### FIFRA SCIENTIFIC ADVISORY PANEL (SAP)

**OPEN MEETING**

**POLLINATOR RISK ASSESSMENT FRAMEWORK**

**DOCKET NUMBER: EPA-HQ-OPP-2012**-**0543 FIFRA SAP WEB SITE http://www.epa.gov/scipoly/sap/**  **OPP Docket Telephone: (703) 305-5805**

**UNITED STATES ENVIRONMENTAL**

**PROTECTION AGENCY**

**CONFERENCE CENTER LOBBY LEVEL ONE POTOMAC YARD (SOUTH BUILDING)**  **2777 SOUTH CRYSTAL DRIVE ARLINGTON, VIRGINIA 22202**  **SEPTEMBER 11 TH** 

### - 14**TH**

### , 2012

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1 **'5)5('-(1.,16-5**Good morning, again. I 

### 2 want to welcome everyone to this FIFRA Scientific Advisory Panel 

### 3 meeting. The topic of this particular meeting is Pollinator 

### 4 Risk Assessment Framework. 

### 5 My name is Fred Jenkins and I am the 

### 6 designated Federal Official for this meeting. Before we get 

### 7 started, I would like to just take a couple of minutes to go 

### 8 over a few administrative items. As a DFO, I serve as a liaison 

### 9 between the panel and the Agency and I am responsible for 

### 10 ensuring that provisions of the Federal Advisory Committee Act 

### 11 are met. 

### 12 I want to extend my thanks to this entire 

### 13 panel for agreeing to serve in this meeting, and to the public 

### 14 as well, for coming to this meeting. The FIFRA SAP is a Federal 

### 15 Advisory Committee that provides independent scientific peer 

### 16 review and advice to the Agency on pesticide issues. 

### 17 It is important to note that the panel only 

### 18 provides the advice and recommendations for EPA, and that all 

### 19 regulatory decision-making and implementation authority remains 

### 20 with the Agency. We have worked with appropriate agency 

### 21 officials to ensure that all appropriate ethics and regulations 

### 22 are satisfied for this meeting. 

### 23 Panel members have been provided their 

### 24 provisions of the Federal Conflict of Interest laws, and each 

### 25 participant has filed a financial disclosure report. I, along 

### 26 with our deputy ethics officer, and in consultation with the

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1 Office of General Counsel, have reviewed these reports to ensure 

### 2 all ethics requirements are met. 

### 3 This meeting provides an opportunity for 

### 4 public comment. We have several people who will be speaking 

### 5 today during the public comment period, which will be tomorrow 

### 6 morning. 

### 7 Now, if you have not made prior arrangements 

### 8 with me to make oral comments and you wish to make an oral 

### 9 comment during the oral comment period tomorrow morning, please 

### 10 let me know or someone else in our SAP staff, who are all 

### 11 situation here to my right. 

### 12 Now, without having made prior arrangements, 

### 13 we ask that you please limit your comments to just five minutes. 

### 14 There is a public docket for this meeting. The docket is 

### 15 listed on the agenda. All background materials and other 

### 16 related documents are available in the dockets and slides of EPA 

### 17 presentations that you will see today. It should be available 

### 18 now. If not, they will be available very shortly. 

### 19 At this point, I want to introduce and extend 

### 20 my thanks to Dr. Daniel Schlenk, to my left, who is serving as 

### 21 Chair to this meeting. He is also the FIFRA SAP Chair. 

### 22 **DR. DANIEL SCHLENK:**Thanks, Fred. Good 

### 23 morning, everyone. Welcome to this particular panel, the 

### 24 Pollinator Risk Assessment Framework. 

### 25 At this point in time, it's customary to go 

### 26 around the room and introduce ourselves. If you wouldn't mind

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1 stating your name, where you're from, and sort of your area of 

### 2 expertise. 

### 3 I'll begin. My name is Dan Schlenk. I am a 

### 4 professor in the Department of Environmental Sciences at the 

### 5 University of California, Riverside. My research expertise is 

### 6 in fate and metabolism of pesticides in aquatic organisms. 

### 7 My job for this particular panel, though, is 

### 8 to keep us on time and get us through all 52 questions before 

### 9 Friday so we can get home on the weekend. 

### 10 So with that, I'll turn it over to Martha. 

### 11 **DR. MARTHA SANDY:**Hello. I'm Martha Sandy. 

### 12 I'm a toxicologist. I'm chief of the toxicology and 

### 13 epidemiology section of the Office of Environmental Health 

### 14 Hazard Assessment. That a department within the California 

### 15 Environmental Protection Agency. 

### 16 **DR. STEPHEN KLAINE:**Good morning. I'm Steve 

### 17 Klaine. I'm a professor at Clemson University and I'm an 

### 18 aquatic ecotoxicologist. 

### 19 **DR. JAMES MCMANAMAN:**Good morning. I'm Jim 

### 20 McManaman. I'm a professor in obstetrics and gynecology at the 

### 21 University of Colorado. I'm vice-Chair for research there. I'm 

### 22 a reproductive biologist. 

### 23 **DR. KENNETH DELCLOS:**I'm Barry Delclos -- or 

### 24 Kenneth Delclos from the FDA's National Center of Toxicological 

### 25 Research. I'm a toxicologist, primarily in reproductive 

### 26 toxicology.

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1 **DR. DAVID TARPY:**My name is David Tarpy. I'm 

### 2 in the Department of Entomology at North Carolina State 

### 3 University and the Extension Apiculturist for North Carolina. 

### 4 **DR. PAUL SCHWAB:**My name is Paul Schwab. I'm 

### 5 an environmental soil chemist at Texas A&M University. 

### 6 **DR. THOMAS POTTER:**Good morning. I'm Thomas 

### 7 Potter. I'm a research chemist with the USDA Agricultural 

### 8 Research Service, Southeast Watershed Laboratory in Tifton, 

### 9 Georgia. And my area of expertise is pesticide environmental 

### 10 fate and transport assessment. 

### 11 **MR. JENS PISTORIUS:**Hello. My name is Jens 

### 12 Pistorius. I'm from Germany and working at the Julius 

### 13 KQhn-Institut, in charge of the risk assessment for pesticides 

### 14 in honey bees for the Investigation Center for Honey Bee Poison 

### 15 Incident and for the governmental research for Germany for 

### 16 pesticide abuse. 

### 17 **DR. JEFF PETTIS:**Good morning. I'm Jeff 

### 18 Pettis. I'm an entomologist and I'm with the USDA Agricultural 

### 19 Research Service and I head the bee research lab here, nearby, 

### 20 in Beltsville, Maryland. 

### 21 **DR. NANCY OSTIGUY:**My name is Nancy Ostiguy. 

### 22 I'm in the entomology department at Penn State. I do a research 

### 23 on honey bee epidemiology. 

### 24 **DR. ROSALIND JAMES:**My name is Rosalind James 

### 25 and my expertise is in bee diseases, a bee pathologist, and I'm 

### 26 director of the Pollinating Insect Research Laboratory in Logan,

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1 Utah, which is part of the USDA Agricultural Research Service. 

### 2 **DR. GREG HUNT:**Good morning. My name is Greg 

### 3 Hunt. I'm at Perdue University. And my area of research is 

### 4 honey bee genomics and behavior. 

### 5 **DR. NINA FEFFERMAN:**Hi. I'm Nina Fefferman. 

### 6 I'm here from Rutgers University. My area of research is 

### 7 applied mathematical modeling of ecological and epidemiological 

### 8 systems. 

### 9 **DR. MAY BERENBAUM:**I'm May Berenbaum, 

### 10 professor and head of the Department of Entomology at the 

### 11 University of Illinois at Urbana-Champaign. And my interest is 

### 12 in insect chemical ecology and chemical mediation with 

### 13 interactions between plants and insects. 

### 14 **DR. DANIEL SCHLENK:**Thank you, everyone. 

### 15 At this point in time, we're going to get introduction and 

### 16 charge from the director of the Office of Pesticide Programs, 

### 17 and that's Dr. Steven Bradbury. 

### 18 **DR. STEPHEN BRADBURY:**Thanks, Dr. Schlenk. 

### 19 Welcome, to all members of the panel to join us for this next 

### 20 week and 52 questions. 

### 21 I think part of the reason you're seeing so 

### 22 many questions is because of the challenge that is before us in 

### 23 undertaking these risk assessments and advancing the science to 

### 24 support our decision-making. I want to give a little bit of 

### 25 background to that before we get started. 

### 26 First, I want to thank you all for all the

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1 work you've already done in reading our White Paper and starting 

### 2 to put together your thoughts on the charge questions, and also 

### 3 thank you for your time this week, and then for the month or two 

### 4 after the meeting when you pull the report together to 

### 5 contribute to these kinds of peer reviews. 

### 6 Sometimes sitting on the other side of the 

### 7 table, I know what it takes. So I greatly appreciate your 

### 8 contribution to the Agency's efforts to advance our scientific 

### 9 undertaking. 

### 10 I also want to thank Fred and members of the 

### 11 SAP panel for all the hard work it takes to get one of these 

### 12 meetings together. 

### 13 The SAP, the Science Advisory Panel, which is 

### 14 created under the FIFRA statute, is a Federal Advisory Committee 

### 15 that provides the Agency with expert advice on the science 

### 16 behind its ecological risk assessments and human health risk 

### 17 assessments for pesticides. This panel is a very important part 

### 18 of the day-to-day operation of our business at EPA, which is 

### 19 undertaking risk assessments for pesticides and making 

### 20 regulatory decisions. And science is the foundation of that 

### 21 decision-making process. 

### 22 So this panel is an important component to 

### 23 that effort, in addition to transparency and openness in public 

### 24 participation, which is critical to our business and as we go 

### 25 forward. 

### 26 So I also want to thank members of the public

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1 who have already provided information to the docket and for the 

### 2 comments that will be coming around tomorrow morning because all 

### 3 that input is very important to your deliberations, as well as 

### 4 the work that we're undertaking. 

### 5 As you know, this week's effort is all around 

### 6 assessing risks of pesticides to pollinators, with the primary 

### 7 focus on honey bees at this stage of our effort. As you know, 

### 8 the White Paper that's been prepared reflects collaboration with 

### 9 Health Canada's Pesticide Management regulatory Authority, as 

### 10 well as California's Department of Pesticide Regulation. 

### 11 And that theme of collaboration, both 

### 12 internationally and nationally, I want to touch upon as I go 

### 13 forward with my remarks. Because clearly, the concepts that we 

### 14 tried to pull together in the White Paper reflect approaches 

### 15 that have been developed, not only with the folks here, but have 

### 16 been enhanced and built from national and international 

### 17 interactions and collaborations with the scientific community 

### 18 and different countries from around the globe. 

### 19 So clearly, what we're presenting here today 

### 20 is a reflection of a lot of work, across a lot of organizations 

### 21 and with a lot of colleagues. Hopefully, we've done justice to 

### 22 the thoughts and ideas that we've put forward in the document 

### 23 which you'll be seeing. 

### 24 As you know, pollinator services are critical 

### 25 to agricultural production and in human food supply and also 

### 26 just the honey itself, and other products that come out of the

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1 hive is an important part of both our nutrition and the commerce 

### 2 of this country. 

### 3 Also, pollinators have been a critical 

### 4 component to the viability of the ecosystems with the 

### 5 pollination that they provide in support of the habitat that we 

### 6 all depend upon. As we know, from the National Academy of 

### 7 Sciences report of 2007, and the yearly USDA reports and other 

### 8 reports around the globe, pollinator decline is a significant 

### 9 issue that we're facing, not only in the United States, but 

### 10 globally. And the causes of pollinator decline are quite 

### 11 complex and clearly not straightforward -everything from habitat 

### 12 modification, habitat loss, nutritional issues, bee management 

### 13 practices, and pesticides all being considered as part of the 

### 14 issues that are facing the viability of honey bees and 

### 15 pollinators in general. 

### 16 At least, from the best we can tell, there 

### 17 isn't a single cause, but clearly, all these stressors are 

### 18 playing some role, to varying degrees, in the decline. Folks in 

### 19 the EPA, as well as our colleagues in PMRA and Cal DPR are 

### 20 taking a hard look at issue and working very diligently with 

### 21 colleagues across the country and the globe in trying to 

### 22 elucidate what these causes can be and how to reconcile these 

### 23 effects. 

### 24 It's clearly a complex issue that's going to 

### 25 take a multi-faceted approach to resolving the issue and working 

### 26 across lots of different organizations and stakeholders to reach

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1 a solution to this problem and try to turn the corner on 

### 2 pollinator populations. 

### 3 In EPA and the Pesticide Program, our primary 

### 4 focus in contributing to this global effort is to try to better 

### 5 understand the role that pesticides may play in pollinator 

### 6 populations and pollinator viability and to advance our risk 

### 7 assessment technique so that we can better elucidate cause and 

### 8 effect and what the potential role of pesticides could be and 

### 9 pollinator honey bee effects, and thereby inform well-reasoned 

### 10 regulatory decisions on the safe use of pesticide products. 

### 11 So our niche in this big challenge is to try 

### 12 to see what we can do to help advance pesticides science and 

### 13 risk assessment techniques. So consistent with this 

### 14 international challenge, and not just the national challenge, 

### 15 we've been working closely with state and federal partners, as 

### 16 we've been moving forward over the years. 

### 17 I think the White Paper reflects an aspect of 

### 18 that collaboration and working with Canada and California's 

### 19 Department of Pesticide Regulation as we pull the White Paper 

### 20 together and some of the approaches from moving forward. 

### 21 Clearly, the White Paper is also drawing upon 

### 22 the deliberations and thoughts from the SETAC Pellston Workshop 

### 23 of a couple of years ago, which brought people together from all 

### 24 over the world to think through the test data requirements and 

### 25 different kinds of risk assessment approaches that could brought 

### 26 to bear.

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1 Also, clearly working with our colleagues at 

### 2 the European Food Safety Authority and getting some of their 

### 3 insights and concepts. You can see that reflected in the 

### 4 aspects of the White Paper, as well as working with our 

### 5 colleagues in OECD, the Organization of Economic Cooperation and 

### 6 Development, where there is an effort across all the member 

### 7 countries of OECD to try to develop harmonized approaches and 

### 8 integrate a variety of techniques and ideas as we go forward. 

### 9 The risk assessment methodology that's 

### 10 described in the White Paper is strongly founded on US EPA's 

### 11 ecological risk assessment framework. And that framework is 

### 12 used, and used quite heavily in the White Paper, but it's used 

### 13 across the agency in how we formulate our risk assessment and 

### 14 undertake the risk assessment. Part of the beauty of that 

### 15 framework is that it helps us integrate not only issues we want 

### 16 to deal with, in terms of pollinators, but our broader 

### 17 components of our ecological risk assessment. So that 

### 18 conceptual model and how it's dealing with fate and transport, 

### 19 for example, of a pesticide, creates a foundation that's also 

### 20 being used in other components of the ecosystems and for other 

### 21 receptors as well. 

### 22 We feel, using a framework, it provides us a 

### 23 solid foundation for what we can do today and what we can do 

### 24 tomorrow. Our first goal by using the framework and all of the 

### 25 various components you'll be hearing about this morning and into 

### 26 the afternoon is that at the end we want to have a risk

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1 characterization component to that risk assessment that provides 

### 2 a transparent, clear, reasonable, and consistent approach to how 

### 3 to interpret the potential risk of pesticides to pollinators, 

### 4 honey bees in particular, in this White Paper. 

### 5 And it's with that clarity of our risk 

### 6 description and risk estimates that we feel it's critical to 

### 7 informing our regulatory decisions as we go forward. 

### 8 Now, clearly, there are a lot of uncertainties 

### 9 that remain and a lot of scientific complexities that aren't 

### 10 resolved yet today. And our risk assessment process in our 

### 11 White Paper hopefully captures what we feel we know something 

### 12 about today, and documents things that we don't know the answers 

### 13 to yet. But our approach is that we feel that we have credible 

### 14 science that we can start moving forward today in advancing our 

### 15 techniques for risk assessments for pollinators, realizing we'll 

### 16 learn more as we go forward. But hopefully we have a foundation 

### 17 upon which we can build upon, so that as new insights and new 

### 18 techniques come to bear, we can fold them into our risk 

### 19 assessment process. 

### 20 So there's always more to learn. There's 

### 21 always more that could be done, but our approach is not to wait 

### 22 for perfection before we can start moving forward and to start 

### 23 to advance our capabilities for undertaking quantitative risk 

### 24 assessments for pollinators. 

### 25 So with that, I want to thank EPA and the PMRA 

### 26 and the California scientists that all contributed to the White

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1 Paper. And I'm really looking forward to the next several days 

### 2 as we go through the presentations and hear your first thoughts 

### 3 on the charge questions. If it's okay with the Chair, I would 

### 4 like to turn it over to Dr. Brady for a little more focused 

### 5 introduction. 

### 6 **DR. DANIEL SCHLENK:**Sure. Before you get 

### 7 started, Dr. Brady, we have some noise coming in on the phone. 

### 8 So if you are on the teleconference, can you please mute your 

### 9 phone? Thank you. 

### 10 Dr. Brady. 

### 11 **DR. DONALD BRADY:**Good morning, members of 

### 12 the panel. I'm Don Brady. I'm the director at the 

### 13 Environmental Fate and Effects Division, here in the Office of 

### 14 Pesticide Programs. 

### 15 Today you will hear presentations designed to 

### 16 solicit your advice on a proposed tiered process for 

### 17 quantitatively evaluating the potential risk to pollinators, 

### 18 utilizing honey bees as surrogates and associated with systemic 

### 19 and non-systemic pesticides. You will hear an overview of a 

### 20 proposed tiered process for quantifying the risks of pesticides 

### 21 to honey bees. 

### 22 As Steve noted, the causes of pollinators are 

### 23 complex and result from many factors. The advice we request 

### 24 today is intended to enhance the ability of EPA, the Pest 

### 25 Management Regulatory Authority in Canada and the California 

### 26 Department of Pesticide Regulation to reliably screen chemicals

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1 for direct and indirect effects on honey bee colonies and on the 

### 2 usefulness of this framework we present for characterizing 

### 3 potential effects to other pollinators as well. 

### 4 I want to thank the panel for the work you 

### 5 will embark on this week. We appreciate and value the advice we 

### 6 receive from this panel. At the outset, I would also like to 

### 7 acknowledge the efforts of the team that prepared this White 

### 8 Paper and presentations. They have worked long and hard to 

### 9 prepare these materials for you. And I want to acknowledge, in 

### 10 addition, the work of Dr. Moshameem (ph), who led the work of 

### 11 the team. 

### 12 Finally, I'm pleased to be here this morning 

### 13 with my colleagues, Mary Mitchell, from Pest Management 

### 14 Regulatory Agency of Canada, and Richard Bireley, of the 

### 15 California Department of Pesticide Regulation, as we initiate 

### 16 this consultation. 

### 17 Their presence today and their participation 

### 18 in the development of the White Paper and the presentations is 

### 19 indicative of the high degree of scientific collaboration among 

### 20 the agencies during the preparation of the White Paper and the 

### 21 presentation and of the Agency's shared commitment to bring the 

### 22 best possible science to bear on this issue. I would like to 

### 23 pass it to Mary if possible. 

### 24 **DR. DANIEL SCHLENK:**Thanks. The panel 

### 25 recognizes Mary Mitchell. 

### 26 **DR. MARY MITCHELL:**Good morning. I'm Mary

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1 Mitchell. I'm Director General of the Environmental Director of 

### 2 Health Canada's Pest Management Regulatory Agency, which 

### 3 effectively makes me Don's counterpart in Canada. 

### 4 I'd like to, first of all, thank the EPA for 

### 5 inviting Canada and California to be part of this event. Like 

### 6 Don, I'd like to acknowledge the collaborative efforts of the 

### 7 U.S. and the Canadian team. They really have worked well 

### 8 together to pool together the proposal you're going to be 

### 9 looking at. 

### 10 I think this is an excellent example of good 

### 11 international collaboration. I strongly believe that 

### 12 international collaboration allows access to the best science 

### 13 available and ultimately results in more robust assessment 

### 14 methods and risk assessments. 

### 15 The international work and pollinator 

### 16 collaboration is part of an ongoing series of collaborations 

### 17 that have been designed to improve and harmonize pesticide data 

### 18 requirements and assessment methods. It's harmonized data needs 

### 19 and assessment methods that make it possible for the U.S., 

### 20 Canada and other countries to share the evaluation of scientific 

### 21 studies, which decreases regulatory burden for both regulated 

### 22 and regulated communities. 

### 23 It also allows the coordinated and efficient 

### 24 pesticide registration decisions which results in better access, 

### 25 faster access to modern, safer pest control products for farmers 

### 26 in all the countries that are involved.

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1 The U.S. and Canada have made it clear that 

### 2 we're highly committed to continuing such international 

### 3 collaboration, whether it be under the auspices of NAFTA, the 

### 4 OECD, or the more recent U.S., Canada Regulatory Cooperation 

### 5 Council that was jointly announced by President Obama and Prime 

### 6 Minister Harper last February. 

### 7 And finally, I would like to thank you, the 

### 8 panel members for agreeing to participate. The breadth and 

### 9 depth of experience in this panel is really amazing. I'm not 

### 10 going to speak to the science; there will be plenty of other 

### 11 people doing that this week. The importance of ensuring 

### 12 pesticide use doesn't impact honey bee health goes without 

### 13 saying -- and I think agencies are really looking forward to 

### 14 your advice on how to further strengthen the proposed framework 

### 15 for characterizing potential effects of pesticides in bees. I'm 

### 16 quite sure this is going to be an interesting week. Thank you. 

### 17 **DR. DANIEL SCHLENK:**Thank you. Next on 

### 18 our agenda is Richard -- is it Bireley? 

### 19 **MR. RICHARD BIRELEY:**Bireley, that is 

### 20 correct. My name is Richard Bireley. I work for the California 

### 21 Department of Pesticide Regulation as an ecotoxicologist. I, 

### 22 too, would like to thank the U.S. EPA for inviting staff from 

### 23 PRMA Canada and the California Department of Pesticide 

### 24 Regulation to participate in this Scientific Advisory Panel to 

### 25 address the potential risks of pesticides to honey bees. 

### 26 The White Paper before you represents a

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1 collaborative effort of OPP's Environmental Fate and Effects 

### 2 Division; Health Canada's Environmental Assessment Directorate 

### 3 and California DPR. 

### 4 Just five years ago, following the review of 

### 5 data submitted as an adverse effects disclosure, staff at Cal 

### 6 DPR realized that the data requirements in 4D CFR were no longer 

### 7 adequate to address the uncertainties posed by pesticides to 

### 8 pollinators. It quickly became clear that this issue had been 

### 9 identified by staff at OPP's Environmental Fate and Effects 

### 10 Division and the Environmental Assessment Directorate at Health 

### 11 Canada. 

### 12 As many of you are aware, California is the 

### 13 home of the largest pollination event in the world, when more 

### 14 than half the bee colonies in the United States are transported 

### 15 to California to pollinate over 800,000 acres of almonds. 

### 16 I believe this international collaboration has 

### 17 resulted in a solid, focused approach that should also increase 

### 18 the confidence of the regulated community in a meaningful 

### 19 product that will meet international data requirements. 

### 20 I would also like to thank each of you serving 

### 21 on the panel for agreeing to vet the proposal and provide input 

### 22 and strengthen it further. I know that Cal DPR staff have found 

### 23 this collaborative effort rewarding, and we all look forward to 

### 24 your input. Thank you. 

### 25 **DR. DANIEL SCHLENK:**Thank you, Mr. 

### 26 Bireley. Okay. Our first presentation this morning will be

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1 from Thomas Moriarty, who is in the Pesticide Reevaluation 

### 2 Division of OPP. 

### 3 **MR. THOMAS MORIARTY:**Thank you, Chairman. 

### 4 And thank you, the FIFRA Scientific Advisory Panel, for allowing 

### 5 me to address you. My name is Tom Moriarty. I'm a risk manager 

### 6 in the Office of Pesticide Programs in the Pesticide 

### 7 Reevaluation Division. 

### 8 During my presentation today, I will be 

### 9 providing an overview of pollinator declines and some of the 

### 10 efforts being conducted here in North American and abroad to 

### 11 refine exposure and effects assessment methods, as well as risk 

### 12 assessment methods for determining the potential impact of 

### 13 pesticides on pollinators and the role the pesticides play and 

### 14 pollinator declines in general. 

### 15 I'll provide a brief overview of the current 

### 16 process used in North America to evaluate the potential effects 

### 17 of pesticides to bees and provide a brief overview of the risk 

### 18 assessment process itself, focusing on problem formulation phase 

### 19 where the protection goals are articulated. 

### 20 Finally, I'll touch upon the extent to which 

### 21 honey bees serve as a surrogate for non-Apis bees and the extent 

### 22 to which a proposed risk assessment process is protective of 

### 23 non-Apis bees. This presentation relates to Charge Questions 1 

### 24 and 3 that deals with the protection goals in non-Apis bees. 

### 25 A number of sources have reported declines in 

### 26 certain pollinator species globally. The 2006 report by the

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1 National Academies of Science indicated decline in some North 

### 2 American pollinators, including North America's most important 

### 3 and productive managed pollinator, the honey bee. 

### 4 However, at the time the NAS report was 

### 5 published, there was insufficient information to determine the 

### 6 cause of these declines. At roughly the same time, some of the 

### 7 declines were being reported in Europe in a report by Biesmeijer 

### 8 et al. in the July 2006 issue of Science. 

### 9 The decline of managed honey bees has been 

### 10 measured by the U.S. Department of Agriculture's National 

### 11 Agricultural Statistics Survey. NASS data indicate that managed 

### 12 honey bee colonies have declined from a peak of approximately 6 

### 13 million colonies in 1947 to roughly 2.8 million colonies in 

### 14 2006. 

### 15 These declines reflect a variety of factors in 

### 16 North American society and agriculture, as well as changes in 

### 17 the way that a NASS itself collect survey data. Despite changes 

### 18 in any survey techniques, data indicates that the overall number 

### 19 of managed colonies had steadily declined over the past 60 

### 20 years. 

### 21 Although not depicted on the graph here, the 

### 22 most recent estimate of 2011 places the number of colonies in 

### 23 the U.S. at roughly 2.5 million. One of the factors affecting 

### 24 honey bee declines has been introductions, spread of Tracheal 

### 25 and Varroa mites. Prior to the introduction of mites, average 

### 26 colony of losses were around five to ten percent; however, since

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1 the introduction of parasitic mites in the U.S. in the 1980s, 

### 2 typical losses of managed honey bee colonies have roughly been 

### 3 about 15 to 21 percent. 

### 4 According to USDA, starting around 2007, 

### 5 average colony loss suddenly increased and ranged between 31 and 

### 6 36 percent. Losses varied among beekeepers, and for some, 

### 7 losses were complete. 

### 8 Some of these losses were marked by sudden 

### 9 loss of adult forage bees, leaving only the queen behind, a few 

### 10 nurse bees, developing brood and ample food reserves. This 

### 11 scenario would lead to the eventual collapse of the colony and 

### 12 was termed Colony Collapse Disorder. 

### 13 While large losses in 2006 and 2007 were 

### 14 attributed to CCD, per se, the contribution of CCD to overall 

### 15 declines in managed colonies appears to be on the decline. 

### 16 Reports of CCD symptoms continue, however, declines in honey bee 

### 17 health are attributed to symptoms such as winterkill, failure to 

### 18 thrive and queen loss. 

### 19 While a number of factors and agents have been 

### 20 hypothesized as potential contributors to CCD and general 

### 21 pollinator declines, at this time, no factor has been identified 

### 22 as single cause. Available science suggests that pollinator 

### 23 declines are a result of multiple factors which may be acting in 

### 24 various combinations. 

### 25 USDA, the lead federal agency on investigating 

### 26 general declines in pollinator health has hypothesized that

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1 losses of managed colonies may be caused by primary stressors 

### 2 such as parasitic mites, poor bee management, nutrition and/or 

### 3 pesticides that may, in turn, cause honey bees to become 

### 4 susceptible to disease and ultimately lead to the collapse of 

### 5 the colony. 

### 6 Research continues to be directed at the range 

### 7 of individual factors, but is also being directed at identifying 

### 8 combinations of stressors that are most strongly associated with 

### 9 pollinator declines. As part of the federal response, a general 

### 10 decline in pollinators, EPA's Office of Pesticide Programs has 

### 11 taken a number of steps to improve the understanding of the role 

### 12 of pesticides and pollinator declines. As part of those 

### 13 efforts, EPA, along with its state and federal partners, has 

### 14 developed a proposed pesticide risk assessment framework for 

### 15 pollinators. 

### 16 Over the past several years, EPA has actually 

### 17 been actively engaged in a number of efforts, both here and 

### 18 abroad, aimed at improving and refining the methods for 

### 19 examining the potential adverse effects of pesticides to bees. 

### 20 In 2009, EPA hosted a USDA-funded meeting to 

### 21 discuss testing protocols for acute and chronic toxicity studies 

### 22 with honey bees. Participants identified uncertainties and 

### 23 limitations of existing protocols and considered refinements to 

### 24 those test protocols. Participants also discussed moving test 

### 25 designs toward more comprehensive and standardized protocols to 

### 26 increase consistency and reproducibility.

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1 In 2011, the Society of Environmental 

### 2 Toxicology and Chemistry, abbreviated as SETAC, sponsored a 

### 3 global Pellston Workshop on pesticide risk assessment for 

### 4 pollinators. The intent was to synthesize the best available 

### 5 science regarding pesticide exposure and effects assessment 

### 6 methods for Apis and non-Apis, and to further harmonize risk 

### 7 assessment approaches among regulatory authorities. 

### 8 The Pellston Workshop had four major themes: 

### 9 1. Exposure Assessment. 

### 10 2. Laboratory Effects Assessment 

### 11 3. Field Effects Assessment 

### 12 4. Integrating Exposure in Effects Data to 

### 13 estimate the likelihood and magnitude of 

### 14 potential effects known as risk assessment. 

### 15 A fifth area of focus was on determining the 

### 16 extent to which honey bee exposure and effects estimates could 

### 17 serve as a reasonable measures with which to evaluate risk to 

### 18 non-Apis bees. The International Commission for Plant Bee 

### 19 Relationships, abbreviated at ICP-BR, which is affiliated with 

### 20 the International Union of Biological Science, draws upon 

### 21 expertise and government industry and academia to inform 

### 22 scientific assessment aimed at bee protection. ICP-BR has been 

### 23 developing methods for testing the toxicity of pesticides to 

### 24 bees and these efforts have informed testing methods supported 

### 25 by the European and Mediterranean Organization for Plant 

### 26 Protection, abbreviated as EPPO, and the Organization for

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1 Economic Cooperation and Development, OECD. 

### 2 The prevention of colony losses, abbreviated 

### 3 as COLOSS, is a global network of scientists, beekeepers and 

### 4 industry members from 55 countries, focused on developing 

### 5 standards for monitoring and research on factors associated with 

### 6 colony losses. 

### 7 The OECD's working group on pesticides has 

### 8 also developed an expert subgroup on pollinators, entitled PEIP, 

### 9 Pesticide Effects on Insect Pollinators workgroup. EPA and PMRA 

### 10 serve as co-chairs to the PEIP workgroup and have been working 

### 11 with other member countries on issues related to pollinator 

### 12 protection. The focus of the PEIP workgroup developed from a 

### 13 2010 OECD member survey on regulatory needs related to 

### 14 pollinators that identified four themes. These themes included 

### 15 advancing and harmonizing the science of risk assessment, 

### 16 developing a source for information on risk management 

### 17 approaches, developing tools to share bee kill incident 

### 18 information, and providing an index of research relevant to 

### 19 pollinator health and protection. 

### 20 Finally, in 2012, the European Food Safety 

### 21 Authority's panel on plant protection products and their 

### 22 residues release a scientific opinion on the development of risk 

### 23 assessment processes for bees, including Apis as well as 

### 24 non-Apis. 

### 25 The opinion examined the potential role of 

### 26 exposure, strengths and limitations of current toxicity testing

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1 methods and provided recommendations on how to improve these 

### 2 methods. Currently, OPP relies on a qualitative process for 

### 3 evaluating the potential hazard of pesticide to honey bees. 

### 4 Typically, the quantified risks to non-target 

### 5 organisms, the Agency relies on point estimates of exposure and 

### 6 effects and the ratio of those estimates, referred to as the 

### 7 risk quotient. Because not every organism can be tested, 

### 8 toxicity testing is typically conducted using surrogate species. 

### 9 10 Selection of a surrogate species is based on 

### 11 the extent to which they are readily available and amenable to 

### 12 testing under laboratory and field conditions. This then 

### 13 reflects the species for which husbandry conditions are well 

### 14 defined. For evaluating potential hazard to terrestrial 

### 15 invertebrates, insect pollinators, toxicity tests are frequently 

### 16 conducted, used in Western or European honey bee. 

### 17 The test listed on this slide will be 

### 18 discussed in greater detail in the presentation to follow. I'll 

### 19 only make several notes on this current tests used in North 

### 20 America to estimate hazard to honey bees. The toxicity tests 

### 21 are typically tiered, where the first tier is an acute toxicity 

### 22 test with young adult bees. 

### 23 According to EPA test requirements, if the 

### 24 acute contact toxicity is less than 11 micrograms per bee and 

### 25 toxicity of residues on foliage may be required. If the 

### 26 toxicity is less than 11 micrograms per bee and there is

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1 evidence of prolonged exposure and/or if there is evidence that 

### 2 bee colonies may be adversely affected, then a field study may 

### 3 be required. 

### 4 Unlike PMRA, EPA guidelines do not currently 

### 5 address acute oral toxicity to capture potential exposure 

### 6 through residues in pollen or nectar that may result from 

### 7 systemic pesticides; nor do current guidelines address the 

### 8 potential toxicity of pesticides to larval bees. However, acute 

### 9 oral toxicity tests are available in the OECD Guidelines, and 

### 10 while acute toxicity tests with larval bees are continuing to 

### 11 evolve, several methods are also currently available. 

### 12 As it will become apparent through the 

### 13 presentations to follow, the closed framework makes use of all 

### 14 the current guideline studies and recommends the addition of 

### 15 certain tests to address areas of uncertainty. 

### 16 The figure above represents the overall risk 

### 17 assessment process. This process is consistent with EPA 

### 18 guidelines and is consistent with a process used by our 

### 19 partners, PMRA and California DPR. It consists of three phases 

### 20 that include problem formulation, analysis, which consists of 

### 21 exposure and effects assessment, and risk characterizations, 

### 22 consisting of risk estimation and risk description. 

### 23 Throughout the assessment process, multiple 

### 24 lines of evidence are synthesized, and although the figure 

### 25 suggests that the process is unidirectional, it is intended to 

### 26 be iterative where exposure and toxicity data reviewed in the

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1 analysis phase may be used to refine the risk hypothesis 

### 2 articulated in the problem formulation phase, or where risk 

### 3 estimates derived in the risk characterization phase may be 

### 4 refined through additional data from the analysis phase. 

### 5 For the remainder of the presentations, 

### 6 detailed elements of this process will be discussed. However, 

### 7 before doing so, I would like to draw your attention to the box 

### 8 at the top left of the diagram, entitled, "Planning." As it 

### 9 indicated, the risk assessment process is intended to be 

### 10 iterative and intended to involve both risk assessor and risk 

### 11 managers. 

### 12 The planning of the risk assessment is 

### 13 intended to occur early in the process as an opportunity for 

### 14 risk managers and risk assessors to articulate management goals 

### 15 and whether there are complexities or issues that may require 

### 16 particular attention. These discussions, in turn, inform 

### 17 decisions regarding resources and scheduling needed to complete 

### 18 the assessment. 

### 19 The presentations today will highlight the 

### 20 tiered nature of the risk assessment process. The initial tier 

### 21 serves as a screen where conservative assumptions about exposure 

### 22 and effects are made, and for those chemicals which do not 

### 23 indicate a minimal risk at the screen level, additional analysis 

### 24 in the form of refinements or higher tiers may be necessary. 

### 25 While the decision to proceed to higher levels 

### 26 of refinement depend on the understanding of both the risk

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1 assessor and risk manager with regard to the management needs. 

### 2 Frequent discussions between the risk assessor and the risk 

### 3 manager often yields opportunities where data or information 

### 4 needs are identified early in the process and it can be 

### 5 communicated to the technical registrant or the appropriate 

### 6 stakeholder. 

### 7 At the top of the figure, the problem 

### 8 formulation phase is identified. The formula is intended to 

### 9 identify the objective of the risk assessment and produce three 

### 10 products. 

### 11 The assessment and measurement endpoints that 

### 12 reflect management goals, conceptual models that describe key 

### 13 relationships between the stressor and the assessment endpoint 

### 14 and the analysis plan. Other presentations you'll hear today 

### 15 will discuss a conceptual model and analysis plan aspects of the 

### 16 problem formulation. But over the next several slides, I'll 

### 17 discuss goals and endpoints and how they are used in the process 

### 18 and their relationship to one another. 

### 19 Ecological risk assessment is typically 

### 20 developed in a risk management context to evaluate human-induced 

### 21 changes that are considered undesirable. Changes often 

### 22 considered undesirable are those that alter important structural 

### 23 or functional characteristics or components of an ecosystem. An 

### 24 evaluation of adverse affects may include consideration of type, 

### 25 intensity and scale of the effects and well as the potential for 

### 26 recovery.

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1 Acceptability of certain adverse effects 

### 2 reflects risk management. It is done so in the light of multiple 

### 3 factors used to weigh both the risks and the benefits of a 

### 4 pesticide. Therefore, it's important that as a risk assessment 

### 5 is being developed, both management goals and science objectors 

### 6 are considered and specificity is given when developing 

### 7 assessment endpoints in conceptual models. 

### 8 Protection goals are statements by a 

### 9 regulatory authority on the desired condition of values of 

### 10 concern. When they are stated at the highest levels, they may 

### 11 be considered generic. U.S. EPA's generic protection goal is to 

### 12 protect human health and the environment. With respect to 

### 13 pesticides, management goals are in part defined by the laws 

### 14 that would provide statutory authority to regulate them. 

### 15 The Federal Insecticide Fungicide and 

### 16 Rodenticide Act directs EPA to regulate pesticides, such that 

### 17 their use does not result in unreasonable adverse effects on man 

### 18 or the environment, taking into account economic, social an 

### 19 environmental costs and benefits from the use of any pesticide. 

### 20 These generic protection goals therefore reflect statutory 

### 21 scientific and societal goals of a regulatory agency. 

### 22 Generic protection goals, however, do not -- 

### 23 may not provide adequate guidance at the risk assessment level. 

### 24 And specific protection goals may be needed which can better 

### 25 direct and inform decisions at that level. Specific protection 

### 26 goals speak more directly to the objectives for an ecological

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1 entity and inform the type of data that is needed; so the 

### 2 specific protection goals therefore must be consistent and 

### 3 support generic protection goals. 

### 4 Consistence with the results of the SETAC 

### 5 Pellston Workshop of specific protection goals with this 

### 6 proposed risk assessment framework include protection of 

### 7 pollination services provided by bees; protection of honey 

### 8 production and other hive products, and the protection of 

### 9 pollinator diversity, in terms of adequate number and diversity 

### 10 species that contribute to the health of the environment. 

### 11 In its recent scientific opinion on the 

### 12 development of risk assessment of plant protection products for 

### 13 bees, the European Food Safety Authority has also identified 

### 14 pollination, hive products and biodiversity as relevant 

### 15 ecosystem services and values to protect. 

### 16 Once protection goals are identified, specific 

### 17 assessment and measurement endpoints can be developed that are 

### 18 consistent with and support the stated protection goals. As 

### 19 defined in the EPA guidelines, assessment endpoints are explicit 

### 20 expressions of the actual environmental values that are to be 

### 21 protected and are operationally defined as an ecological entity 

### 22 and its attributes. 

### 23 The ability of assessment endpoints to support 

### 24 risk management decisions is dictated by whether or not there 

### 25 are measurable characteristics that adequately represent or 

### 26 reflect management goals. EPA has identified three criteria for

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1 determining the selection of assessment endpoints and it 

### 2 included whether they are ecologically relevant, susceptible to 

### 3 the known or potential stressor, and what's relevant to the 

### 4 management goals. 

### 5 Assessment endpoints are then specifically 

### 6 informed by the data generated for the risk assessment. Both 

### 7 effects data and exposure data provides specific information as 

### 8 measurement endpoints than inform one or more of the assessment 

### 9 endpoints. 

### 10 When measurement endpoints are appropriately 

### 11 linked to assessment endpoints and specific protection goals, 

### 12 they therefore also support generic protection goals. This 

### 13 slide is Table 1 of the White Paper and illustrates both 

### 14 examples of and the relationship between measurement endpoints, 

### 15 assessment endpoints and specific protection goals. 

### 16 The specific protection goals identified in 

### 17 the table are then consistent with the generic protection goals 

### 18 of protecting human health and the environment. The measurement 

### 19 endpoints identified in the table are not intended to be 

### 20 exhaustive, but only serve as examples. For the remaining 

### 21 presentations, a range of measurement endpoints will be 

### 22 discussed in the context of the pro's risk assessment framework. 

### 23 24 In conclusion, the presentations you will hear 

### 25 today will layout a proposed risk assessment framework that is 

### 26 tiered, iterative and flexible, allowing for multiple lines of

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1 evidence to be considered and refinements to be made. In this 

### 2 way, the proposed framework is sensitive with respect to the 

### 3 resources needed to complete a risk assessment. 

### 4 The primary focus of the proposed framework is 

### 5 for evaluating the potential for adverse effects to honey bees 

### 6 and also uses a honey bee as a surrogate for other non-Apis 

### 7 bees. The extent to which any species is a reasonable surrogate 

### 8 for other species, involves considerable uncertainty, unless 

### 9 sufficient data are available to put differences in 

### 10 relationships into context. 

### 11 Uncertainties reflect differences in life 

### 12 biologies and histories that can impact both exposure and 

### 13 effects. Despite the potential differences among bee species, 

### 14 it is believed that the proposed risk assessment process with 

### 15 honey bees provides information on non-Apis bees as well. At 

### 16 lower tiers, for example, where effects to individual bees is 

### 17 tested, exposure routes considered, i.e. contact and ingestion 

### 18 are shared by both honey bees and non-Apis bees alike. 

### 19 Until guideline studies with non-Apis bee 

### 20 species are better developed and available for regulatory 

### 21 purposes, the Agency is asking the panel the extent to which it 

### 22 believes the proposed processed is informative of a potential 

### 23 pesticide risk to non-Apis species. 

### 24 Finally, EPA and its partners seek the opinion 

### 25 of the panel regarding the extent to which the proposed risk 

### 26 assessment framework supports the identified protection goals

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1 and the extent to which biodiversity, identified as a protection 

### 2 goal, is supported by the use of a honey bee, insofar as it is a 

### 3 reasonable surrogate for non-Apis bees. 

### 4 Thank you for your time. And if there are no 

### 5 questions, I would like to allow Mr. Sappington to move into his 

### 6 presentation on conceptual models. 

### 7 **DR. DANIEL SCHLENK:**I think we do have some 

### 8 questions. Again, this is a general overview. So if you could 

### 9 hold your questions to more of the general framework, I believe 

### 10 future speakers will have very specific details on the process. 

### 11 So if you have questions related to the overall objectives, 

### 12 then, yeah. Dr. James. 

### 13 **DR. ROSALIND JAMES:**I have a simple question. 

### 14 You talked about management goals. The word "management" has a 

### 15 lot of meanings to me. Are you talking about pest management 

### 16 goals in your model framework? 

### 17 **MR. THOMAS MORIARTY:**Risk management goals, 

### 18 balancing between the risks and benefits. 

### 19 **DR. DANIEL SCHLENK:**That was Mr. Moriarty. 

### 20 Let me just remind the speakers that anytime you respond, can 

### 21 you state your name real quick beforehand so that we can get 

### 22 that on the written record. Thank you. 

### 23 Any other questions? 

### 24 (No response.) 

### 25 Okay. Mr. Sappington, do you want to take the 

### 26 reins for the next presentation? We can get that loaded.

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1 **MR. KEITH SAPPINGTON:**Hello. My name is 

### 2 Keith Sappington. I'm a biologist and senior science advisor in 

### 3 the ecological Environmental Fate and Effects Division within 

### 4 the Office of Pesticide Programs. 

### 5 I appreciate this opportunity to address the 

### 6 FIFRA Scientific Advisory Panel on this important topic, and I 

### 7 look forward to your feedback to help us improve our process for 

### 8 pesticide risk assessment to bees. 

### 9 This morning I'll be summarizing two aspects 

### 10 of the White Paper in back-to-back presentations. In the first 

### 11 presentation, I will describe a set of generic conceptual models 

### 12 we are proposing for assessing pesticide risk to honey bees. 

### 13 Details of these generic conceptual models are 

### 14 found in Section II of the White Paper. Immediately following 

### 15 this presentation I will summarize our proposed decision 

### 16 framework for assessing risks of pesticides to honey bees. And 

### 17 this is also found in Section II of the White Paper. 

### 18 With respect to conceptual models, I will 

### 19 first provide a brief overview of the use of conceptual models 

### 20 and ecological risk assessment, and then I'll describe the 

### 21 context and the purpose of these models as we apply them in 

### 22 ecological risk assessment of pesticides to honey bees. 

### 23 Following this introduction, I'll then 

### 24 describe the generic conceptual models that are specific to four 

### 25 common application methods. And these include foliar spray 

### 26 applications for both systemic and non-systemic pesticides, soil

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1 application for systemics, and seed treatment application for 

### 2 systemic pesticides. 

### 3 We present a fifth conceptual model in the 

### 4 White Paper for trunk drench and tree injection, but I will not 

### 5 be presenting that this morning. 

### 6 The figure on the right of this slide 

### 7 illustrates the components of problem formulation phase and it 

### 8 depicts, visually, how the selection of assessment endpoints and 

### 9 conceptual models are used to inform the analysis plan of the 

### 10 risk assessment. 

### 11 As defined in EPA's ecological risk assessment 

### 12 guidelines, a conceptual model is a written and visual 

### 13 description of the hypothesized relationships between the 

### 14 ecological entities and the stressors of concern. In the 

### 15 conceptual models I'll be presenting today, the ecological 

### 16 entities, or honey bees, and the stressor of concern is the 

### 17 pesticide application, and this includes any degradates that are 

### 18 of toxicological concern. 

### 19 The written portion of a conceptual model 

### 20 includes a set of risk hypotheses which are statements regarding 

### 21 the purported relationships between the entities and the 

### 22 stressor. The visual portion of the conceptual model -- which I 

### 23 will be presenting shortly -- are diagrams of these purported 

### 24 relationships. 

### 25 In the context of ecological risk assessment, 

### 26 conceptual models are useful tools for communicating to risk

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1 managers, risk assessors and the public of how we believe 

### 2 exposure to pesticides and their associated effects are likely 

### 3 to occur. 

### 4 With that said, there is not a single standard 

### 5 format for depicting a conceptual model diagram. They can vary 

### 6 widely in their level of detail, from simple, highly aggregated 

### 7 diagrams to ones that are very complex and highly disaggregated. 

### 8 9 In addition, the graphical format can vary, 

### 10 for example, from the more traditional box and arrow diagrams, 

### 11 to ones that are pictorial in their representation. But 

### 12 regardless of format, the general structure of conceptual models 

### 13 is generally similar and, as indicated on the bottom of this 

### 14 slide, represents a cascade of events which begins at the 

### 15 stressor source and its relevant exposure routes, and then to 

### 16 receptors of concern and their responses, and ultimately to 

### 17 changes in the attributes or the assessment endpoints 

### 18 indentified in the risk assessment. 

### 19 The generic conceptual models I'll be 

### 20 presenting this morning are consistent with the format typically 

### 21 used in EPA pesticide risk assessments. Before presenting the 

### 22 conceptual models, I first would like to make several points 

### 23 regarding their development and intended use in risk assessment. 

### 24 25 First, the generic conceptual models are 

### 26 called generic because they're intended to be used as a starting

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1 point for pesticide risk assessors. And therefore, are expected 

### 2 to be modified, where necessary, depending on the circumstances 

### 3 of that specific risk assessment. 

### 4 Second, the conceptual models presented here 

### 5 are not comprehensive of all pesticide uses. Rather, they 

### 6 represent several major use categories that are expected to 

### 7 result in some exposure to honey bees. 

### 8 Lastly, in addition to being organized around 

### 9 different pesticide use patterns, the models also consider some 

### 10 differences in chemical characteristics, and in particular, the 

### 11 systemicity of a pesticide, which, in turn, impacts the nature 

### 12 of exposure to honey bees. 

### 13 In describing the generic conceptual models, I 

### 14 first plan to go through in detail the conceptual model for 

### 15 foliar applications of non-systemic pesticides here and then 

### 16 I'll highlight the main differences for the other three 

### 17 conceptual models that we have identified. 

### 18 As shown in bold, on the left, these 

### 19 conceptual models begin at the top, with the stressor of concern 

### 20 and continue downward by illustrating the sources of exposure, 

### 21 the related exposure media, receptors of concern and changes in 

### 22 the attribute or assessment endpoints. 

### 23 In this case, the stressor of concern is a 

### 24 non-systemic pesticide application -- and again, including any 

### 25 of its degradates -- that is applied to a crop foliage, be it 

### 26 ground or application methods. As shown in these boxes, the

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1 exposure sources considered in this model include deposition of 

### 2 spray droplets directly onto bees, plants, soil, and surface 

### 3 water, either on the treated site or via drift, to adjacent 

### 4 sites. 

### 5 From these exposure sources, resulting 

### 6 pesticide residues in various environmental media are 

### 7 considered, including residues on plant surfaces, in soil, 

### 8 pollen and nectar, and in surface water, which can result from 

### 9 spray drift and pesticide runoff/erosion, shown in blue. 

### 10 You'll notice that the dashed line surrounding 

### 11 the residues in the soil and surface water boxes. This 

### 12 indicates that we believe these exposure pathways are less 

### 13 important to honey bees compared to other exposure routes 

### 14 depicted. Additional information supporting this distinction 

### 15 will be presented later in a presentation by Ms. Christina 

### 16 Wendel. 

### 17 The nature of pesticide exposure is then 

### 18 depicted by the lines connecting the pesticide residues and 

### 19 environmental media to the receptors. Worker bees foraging for 

### 20 food and water sources constitute the primary receptor of 

### 21 concern outside of the hive; although direct exposure to drones 

### 22 and the queen is also possible during mating and orientation 

### 23 flights. 

### 24 Primary routes of exposure outside of the hive 

### 25 that we have included include dermal uptake of pesticide, via 

### 26 spray droplets and foliar residues, inhalation -- which is

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1 applicable to more volatile pesticides, primarily -- ingestion 

### 2 of pesticides absorbed onto pollen and nectar. 

### 3 Secondary routes of exposure of lessor concern 

### 4 for honey bees include contact with contaminated soil and 

### 5 surface water. Another secondary route of exposure not depicted 

### 6 here is contaminated dew on plant surfaces. Exposure from 

### 7 contaminated dew droplets is discussed in detail in a later 

### 8 presentation. 

### 9 We note, however, that exposure from pesticide 

### 10 residues in soil is likely to be more important for some 

### 11 non-Apis bee species. For example, ground nesting bees that 

### 12 collect and process soil for nest construction. 

### 13 From here, exposure to bees inside the hive is 

### 14 possible, via direct contact, ingestion and processing of 

### 15 contaminated pollen and nectar brought into the hive by the 

### 16 workers. Ingestion of honey and as well as processing of wax 

### 17 and propolis during comb production and brood raring activities. 

### 18 19 Exposure of hive bees to pesticide is also 

### 20 possible from contaminated water used for consumption, honey 

### 21 production, and evaporative cooling. And this pathway is 

### 22 further described in a later presentation. 

### 23 Subsequently, bee brood may be exposed to 

### 24 pesticides via ingestion of brood provisions that includes brood 

### 25 food, honey, and processed pollen, wax and propolis. Inside the 

### 26 hive, queens are expected to be exposed primarily through

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1 contact with comb material and consumption of royal jelly. 

### 2 Lastly, the relationships between the exposure 

### 3 of different bee castes and changes in attributes or assessment 

### 4 endpoints are shown with these arrows and text boxes. The text 

### 5 in bold in these boxes represent either the assessment endpoints 

### 6 or in a case to pollinator biodiversity, the protection goal. 

### 7 Example of measurement endpoints are shown below at each of the 

### 8 assessment endpoints. 

### 9 We note that these arrows collapse many 

### 10 complex chemical, biological and toxicological processes that 

### 11 collectively determine the responses of individual bees and the 

### 12 entire colony to a given pesticide exposure. For example, the 

### 13 populations size and stability of colonies may not only be 

### 14 impacted by a direct loss in forager bees due to mortality, but 

### 15 also by other factors, including, but not limited to reduced 

### 16 foraging success and associated reduction in food reserves, 

### 17 insufficient workers to maintain hive temperatures during 

### 18 winter, and a reduction in homing success. 

### 19 For systemic foliar applied pesticides, the 

### 20 generic conceptual model is identical to that described 

### 21 previously for non-systemic pesticides, except that it includes 

### 22 pesticide uptake and translocation to the plant tissues. And I 

### 23 show this in red in the arrows in this figure. 

### 24 Thus, for systemic pesticides that are applied 

### 25 to foliage via spray method, a portion of the residues in pollen 

### 26 and nectar may result from pesticide deposition onto these

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1 tissues in addition to a portion that may be subsequently 

### 2 transported to these tissues via translocation. 

### 3 The remainder of this conceptual model from 

### 4 the receptors downward is the same as described previously for 

### 5 non-systemic foliarly applied pesticides. For application of 

### 6 systemic pesticides directly to bare soil either by granular 

### 7 spray or soil incorporation methods, the dominate route of 

### 8 exposure we have depicted for honey bees begins with pesticides 

### 9 uptake and translocation within the developing plant. 

### 10 As with the foliarly-applied systemic 

### 11 pesticides, the foraging bees may then be exposed from ingestion 

### 12 and contact with residues in pollen, nectar, plant exudates, and 

### 13 honey dew. It is assumed that direct exposure of foraging bees 

### 14 with pesticide spray droplets would not be a primary exposure 

### 15 route of concern because soil applications would be presumed to 

### 16 occur prior to planting. 

### 17 Furthermore, spray applications to bare soil 

### 18 typically involve a coarse droplet spectrum which would not 

### 19 likely to be subject to extensive offsite drift. However, if 

### 20 offsite drift of spray droplets was a concern, or if there was 

### 21 attractive foliage in the field at the time of the soil 

### 22 application, then these exposure routes would be addressed as 

### 23 described previously for the foliar spray conceptual model. 

### 24 Runoff of a pesticide into surface water is 

### 25 another potential route of concern, but this is expected to be 

### 26 minor relative to exposure from residues in pollen and nectar.

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1 Again, the remainder of the conceptual model 

### 2 from the receptors downward is the same as described previously. 

### 3 Exposure of honey bees to pesticides via seed treatment is 

### 4 depicted here as a result of two major sources: offsite drift of 

### 5 abraded seed coat dust and from pesticide residues in plants 

### 6 that result from pesticide translocation to pollen, nectar, 

### 7 plant exudates and honeydew. 

### 8 Subsequent exposure of bees to pesticides in 

### 9 these matrices follows the same pathways as depicted earlier for 

### 10 soil application of systemic pesticides. For drift of abraded 

### 11 seed coat dust, foraging bees may be exposed by directly 

### 12 intercepting dust particles containing the pesticide during 

### 13 flight and subsequent dermal or respiratory uptake. It is also 

### 14 possible for contaminated dust to deposit on other environmental 

### 15 media, including soils, surface water, and plant tissues. 

### 16 Generally speaking, these routes are likely to 

### 17 result in lower exposures compared to direct contact with seed 

### 18 dust or contaminated pollen and nectar from the treated crop. 

### 19 We note, however, that exposure to pesticides 

### 20 from deposition of contaminated dust particles onto flowing 

### 21 plants may be important, particularly if these plants are 

### 22 located in close proximity to the seed planting operations. 

### 23 Again, the bottom half is the same as I described earlier. 

### 24 So to conclude, the generic conceptual models 

### 25 for pesticide exposure and effects on honey bees presented here 

### 26 and in the White Paper, are intended to reflect only several

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1 major pesticide use categories. They also are intended to serve 

### 2 as a basis for consideration in a forthcoming pesticide risk 

### 3 assessment involving honey bees, and are expected to be modified 

### 4 when circumstances warrant. 

### 5 As indicated in Charge Question Number 2, we 

### 6 are seeking feedback on these models, in terms of their overall 

### 7 accuracy and the extent to which their implementation is 

### 8 consistent with the proposed assessment endpoints and management 

### 9 goals. 

### 10 At this time, myself and the team would 

### 11 address any clarifying question on the generic conceptual 

### 12 models. And after that, I will present on our proposed decision 

### 13 frameworks. Thank you very much. 

### 14 **DR. DANIEL SCHLENK:**Thank you. Okay, any 

### 15 questions of clarifications. Dr. Potter? 

### 16 **DR. THOMAS POTTER:**I have a question about 

### 17 your category of foliar-applied pesticide. Does that 

### 18 essentially include all non-systemic broadcast applied active 

### 19 ingredients? 

### 20 **MR. KEITH SAPPINGTON:**For the first 

### 21 conceptual model I presented, that's for non-systemic 

### 22 foliar-applied sprays. 

### 23 **DR. THOMAS POTTER:**But does it have to be 

### 24 foliar applied is, I guess, where I'm looking for an answer on 

### 25 that? 

### 26 **MR. KEITH SAPPINGTON:**Well, we distinguish

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1 between foliar and soil applied. So the presumption is that it 

### 2 is being applied to foliage in that conceptual model. 

### 3 **DR. THOMAS POTTER:**But it seems to leave out 

### 4 a large category of active ingredients that one presumes 

### 5 assessments being made regarding risks, pre-emergent herbicides 

### 6 would be one category. 

### 7 **MR. KEITH SAPPINGTON:**Right. Those would, I 

### 8 think, be covered in the soil application methods; the 

### 9 conceptual model for soil application. 

### 10 **DR. THOMAS POTTER:**I guess what I saw for 

### 11 soil application, though, was systemics and not, you know, non - 

### 12 again, non-systemics are what we're talking about here. Again, 

### 13 I'm just looking for clarification on this as to where those 

### 14 categories of chemicals that are soil applied, that are 

### 15 non-systemic, where do they fall into these four different 

### 16 categories? 

### 17 **MR. KEITH SAPPINGTON:**For non-systemic 

### 18 soil-applied pesticides, you would be primarily concerned about 

### 19 the contact exposure; although, again, if they're applied 

### 20 pre-emergence to bare soil then the exposure to bees foraging in 

### 21 that particular field site would be presumed to be rather low. 

### 22 If there were foliage present then it would follow the foliar 

### 23 conceptual model. Or, if there was a reason to expect drifts 

### 24 offsite from that, then it would follow the foliar conceptual 

### 25 model framework. 

### 26 **DR. THOMAS POTTER:**So again, just to be

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1 clear, all non-systemic active ingredients, which would be 

### 2 broadcasts applied, would fall into that first schematic that 

### 3 you showed? 

### 4 **MR. KEITH SAPPINGTON:**I think that would be 

### 5 the closest one that would cover them. Now we could split those 

### 6 out as we refine these conceptual models to make that more 

### 7 clear. But we focused on the systemics because of the focus on 

### 8 the uptake in translocation in the plants and exposure through 

### 9 pollen and nectar. But we can break that out to make that 

### 10 clearer. Thank you. 

### 11 **DR. DANIEL SCHLENK:**Any others? Yes, Dr. 

### 12 McManaman. 

### 13 **DR. JAMES MCMANAMAN:**I see that larvae left 

### 14 out of your model here, it doesn't look like they're present in 

### 15 there. Is that to simply the model or is that because you have 

### 16 in mind a different kind of model for larvae? 

### 17 **MR. KEITH SAPPINGTON:**The larvae would be 

### 18 represented towards the bottom of the conceptual model as part 

### 19 of the brood in the hive. That's where we would be expecting 

### 20 their exposure for honey bees. 

### 21 **DR. JAMES MCMANAMAN:**So you make a 

### 22 distinction between the brood provisions and provisions for the 

### 23 queen, in terms of royal jelly. But my understanding is that 

### 24 oftentimes early on, the bees receive the same kind of food 

### 25 until it's distinguished between who's going to become queen or 

### 26 not. Is that again left out due to simplification?

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1 **MR. KEITH SAPPINGTON:**Let me see if I can 

### 2 understand the question first. So the question is, is direct 

### 3 exposure from pollen and nectar to bee brood as opposed to its 

### 4 processed pollen and nectar and how is that represented in the 

### 5 conceptual model. Do I understand it correctly? 

### 6 **DR. JAMES MCMANAMAN:**Yes. 

### 7 **MR. KEITH SAPPINGTON:**Yes. My understanding 

### 8 is that was left out just for simplicity reasons. I mean, at 

### 9 some point you get to have so many hours and lines directed. My 

### 10 understanding is that -- and we've investigated and I believe 

### 11 Ms. Kris Garber will be presenting more information about the 

### 12 consumption of larvae later today -- but we've investigated the 

### 13 extent to which the larvae were consuming or may consume pollen 

### 14 directly versus the process pollen. 

### 15 **DR. JAMES MCMANAMAN:**So the Agency's view is 

### 16 that larvae are included in this model system and the details 

### 17 may -- 

### 18 **MR. KEITH SAPPINGTON:**Absolutely. 

### 19 **DR. JAMES MCMANAMAN:**-- vary from adults. 

### 20 **MR. KEITH SAPPINGTON:**Absolutely. 

### 21 **MR. JAMES MCMANAMAN:**Okay. 

### 22 **DR. DANIEL SCHLENK:**Any other questions. 

### 23 Clarification? Dr. James? 

### 24 **DR. ROSALIND JAMES:**This is Rosalind James. 

### 25 I think this is the appropriate place to ask this question. 

### 26 When honey is made, nectar is collected and then a significant

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1 proportion of the water is evaporated off and it becomes 

### 2 concentrated. Is that included in your analysis at all? 

### 3 **MR. KEITH SAPPINGTON:**We have a separate 

### 4 presentation this afternoon that looks at the whole issue of 

### 5 contaminated surface water and its relative importance, compared 

### 6 to these other aspects. 

### 7 **MS. ROSALIND JAMES:**I'm actually talking 

### 8 about the nectar itself. So the honey is made from nectar and 

### 9 then the nectar is predominately water. But it's the nectar -- 

### 10 I'm not talking about collected water. And then to make honey, 

### 11 the bees evaporate off the -- 

### 12 **MR. KEITH SAPPINGTON:**Right. 

### 13 **MS. ROSALIND JAMES:**I mean, that's processing 

### 14 of the nectar. But is the fact that you're concentrating the 

### 15 nectar down included in your risk assessment? It could change 

### 16 the exposure level. 

### 17 **MR. KEITH SAPPINGTON:**That's right. There 

### 18 are basically two competing processes, potentially, as we 

### 19 understand them. One would be degradation of the pesticide over 

### 20 time. And then conceptually, there could be some concentration 

### 21 as the honey is formed. I personally am not aware of data on 

### 22 that concentration phase, but those processes would be, I think, 

### 23 in competition with one another. 

### 24 **DR. THOMAS STEEGER:**I would like to address 

### 25 Dr. James's question. As you will learn from the presentation 

### 26 from Kris Garber later on today, the concentration of residues

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1 that could potentially take place as honey is being processed 

### 2 into the colony is accounted for in our exposure modeling 

### 3 because it's expressed on a sugar basis, as opposed to a water 

### 4 basis. So hopefully after you hear that presentation you'll 

### 5 have a better understanding of how we've accounted for that 

### 6 potential mechanism of exposure. 

### 7 **DR. DANIEL SCHLENK:**Thank you, Dr. Steeger. 

### 8 Any other questions? Dr. Fefferman. 

### 9 **DR. NINA FEFFERMAN:**My guess is that that the 

### 10 answer to this will be addressed in a later portion. But I was 

### 11 wondering, in the conceptual modeling where you felt that the 

### 12 layer for how the impact might propagate throughout a colony, it 

### 13 looks to me like everything that you've discussed is sort of 

### 14 exposure to individual bees and how that might get to individual 

### 15 bees. Where do you see that layer fitting? 

### 16 **MR. KEITH SAPPINGTON:**I agree. As I went 

### 17 through the conceptual model I noted that those lines would 

### 18 connect the exposure media to the receptors and ultimately to 

### 19 the changes in the attributes, collapse a lot of information 

### 20 down. One aspect that I've thought out, personally, you know, 

### 21 there's no reason that a conceptual model necessarily has to fit 

### 22 on one page. 

### 23 So we could have a conceptual model, much like 

### 24 this, that addresses the exposure pathways and then a separate 

### 25 one that lays out the various ways in which the response is to 

### 26 that given exposure, could then have affects on the hive. And

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1 that could be very useful for the planning phase of a risk 

### 2 assessment. Thank you for that comment. 

### 3 **DR. NINA FEFFERMAN:**Is that currently in some 

### 4 section somewhere for consideration or is that something that 

### 5 really might be flushed out better in the conceptual model 

### 6 section but separately, as you proposed? 

### 7 **MR. KEITH SAPPINGTON:**We discuss, texturally, 

### 8 in the White Paper these various ways in which the colony 

### 9 responses could be manifested, but they're not visually 

### 10 presented, currently. 

### 11 **DR. THOMAS STEEGER:**I would like to add to 

### 12 Mr. Sappington's response by saying that as you will learn in a 

### 13 later presentation, it's not captured in the conceptual model, 

### 14 but we evaluate multiple lines of evidence to estimate, not just 

### 15 risk to individual bees, but to the colony as a whole. And part 

### 16 of that process we will be asking for SAP input on, is the use 

### 17 of colony level modeling in addition to the advanced or more 

### 18 refined testing that is conducted at the semi-field and full 

### 19 field level that look at whole colony effects. 

### 20 **DR. DANIEL SCHLENK:**Okay. Any other 

### 21 questions before we break? 

### 22 (No response.) 

### 23 All right. Thanks. We're going to go by this 

### 24 clock on the wall here. Let's be back at 10:35. 

### 25 (Brief recess.) 

### 26 **DR. DANIEL SCHLENK:**Okay. Thanks. Again,

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1 let me just reiterate, if you're on the phone, please mute your 

### 2 line at this time so we don't hear you during the presentations. 

### 3 4 Before we go to our next presentations, I 

### 5 believe there were some questions, some overall framework 

### 6 questions that were not asked the last time, for you, Keith, if 

### 7 that's okay. I believe Dr. Fefferman has questions, please. 

### 8 **DR. NINA FEFFERMAN:**So forgive me, these 

### 9 don't actually address the specific conceptual models you were 

### 10 putting forth because I think those are really quite complete 

### 11 and lovely, and this is more a structural, as we consider the 

### 12 framework of risk assessment itself type of question. 

### 13 In the types of risk assessment frameworks 

### 14 that I'm used to considering, sort of the very first step is 

### 15 which things are going to be useful to measure and which things 

### 16 are going to inform choices and how do we balance those. And I 

### 17 feel sort of as though we skipped past that to here are the 

### 18 things we're definitely measuring and here are the conceptual 

### 19 models and how those measurement might play out in terms of the 

### 20 risks associated with those particular aspects. 

### 21 Has there been a stage of evaluation about 

### 22 some of these? I mean, I know we're not discussing necessarily 

### 23 the benefits of some of the pesticide use, but some of the risks 

### 24 to colonies, to bees, has there been a stage of thinking 

### 25 relative impact, types of measurements available, when we've got 

### 26 diminishing returns from better measurement, that kind of thing?

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1 **DR. THOMAS STEEGER:**I think what we're trying 

### 2 to depict here is the generalized process that the agency -- it 

### 3 plans on or proposes to use for honey bees and to the extent 

### 4 that honey bees serve as surrogates for non-Apis bees as well. 

### 5 And this process has to rely, at least initially, as with all 

### 6 taxa, a screening level that is relatively generic and looks at 

### 7 very broad categories of effects to see where resources really 

### 8 need to be expended to focus in on where risk, at this very 

### 9 broad level, cannot be dismissed. 

### 10 So when you talk about the nuances about 

### 11 colony level effects -- 

### 12 **DR. NINA FEFFERMAN:**Forgive me. I don't 

### 13 actually mean nuances of particular effects; I mean actually 

### 14 going more abstract rather than more specific. 

### 15 There seems to me to be layers of very 

### 16 specific and then we measure this type of pesticide application 

### 17 and how much gets into food supply. You know, that all seems to 

### 18 be to me to be a very specific measurement as opposed to much 

### 19 broader things like, well, what are the toxicological effects at 

### 20 this layer, as opposed to we measure this flow path or something 

### 21 like that. 

### 22 Has that kind of assessment of a more abstract 

### 23 flow process for risk been accomplished already? 

### 24 **DR. THOMAS STEEGER:**I think -- and correct me 

### 25 if I'm wrong -- you're getting at this issue that the agency 

### 26 routinely wrestles with and it's this concept of a adverse

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1 outcome pathway and how do you transition from one level of 

### 2 biological organization to the next. 

### 3 I think that we're looking, as this process 

### 4 evolves, to help develop a more standardized approach for making 

### 5 those linkages. That's one of the difficulties that we have in 

### 6 the use of many of the sublethal endpoints that have been 

### 7 identified in some of the bee studies that are reported in the 

### 8 open literature. That's a challenge. 

### 9 I'll say that we don't have all the answers at 

### 10 this point. But for the frank effects that we traditionally use 

### 11 of acute lethality and survival and reproduction. Those are the 

### 12 affects that right now are what the assessment endpoints are 

### 13 based on and the measurement endpoints that we plan to quantify 

### 14 as we develop this process further. 

### 15 But developing those linkages that would 

### 16 better enable us to move from one level of biological 

### 17 organization to the next, you're absolutely right. That's what 

### 18 the ultimate endgame is. Have they been developed yet? No. 

### 19 But the science is still evolving. 

### 20 **DR. NINA FEFFERMAN:**Okay. Great. Thanks. 

### 21 So you're thinking of that as something that comes out of this 

### 22 level as opposed to something that is done before we get down to 

### 23 the drill down level of these discussions? 

### 24 **DR. THOMAS STEEGER:**I think that the process 

### 25 that we're using to refine risk assessment where we identify 

### 26 chemicals that there appears to be risk that can't be

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1 discountable at the screening level, that we move to higher tier 

### 2 testing at the whole colony level to try to detect whether under 

### 3 actual use conditions is the colony going to be able to survive, 

### 4 either in the short-term or in the long-term? 

### 5 Having very bright lines at the full colony 

### 6 level is difficult to establish at this point because these 

### 7 nuances that I mentioned earlier, we don't really have a clear 

### 8 understanding of how much of an impact can a colony sustain or a 

### 9 population of bees can sustain, over long periods of time. 

### 10 I think that that is something that still has 

### 11 to evolve, but our screening level process is geared towards 

### 12 trying to do that type of refinement, but at this point, it 

### 13 would be a qualitative assessment to determine is the colony 

### 14 going to make it or not. 

### 15 **DR. NINA FEFFERMAN:**Thank you. I think your 

### 16 comments directly address my question. So I guess my take home 

### 17 from this might be that on some level, we might consider, as 

### 18 we're talking, that some of this might be putting the cart 

### 19 before the horse, where as if, when we get to that qualitative 

### 20 level, we discover that some of these aspects are not having any 

### 21 impact than the careful ability to measure the local impact, may 

### 22 not help us build up to that level. So that's great. Thank 

### 23 you. I just want to get that on the record. Thanks. 

### 24 **DR. THOMAS STEEEGER:**I think you put that 

### 25 very well. 

### 26 **DR. DANIEL SCHLENK:**Great. Thanks a lot

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1 for getting that clarified. Okay. Our next speaker is 

### 2 Christina Wendel. Oh, sorry. You have a second one? Oh, 

### 3 sorry. Keith Sappington, please. 

### 4 **MR. KEITH SAPPINGTON:**Thank you. In my 

### 5 second presentation, I will describe two risk assessment 

### 6 decision frameworks that we've proposed to use as guide for 

### 7 assessing pesticide risk to honey bees. Here, I will only 

### 8 summarize the proposed decision frameworks and their components 

### 9 and detail specific to each of these components will be 

### 10 presented in subsequent presentations. 

### 11 In describing the proposed decision 

### 12 frameworks, I'll first begin by summarizing some of their key 

### 13 attributes which shape their development. I will then describe 

### 14 the proposed decision frameworks for pesticides that are applied 

### 15 via foliar sprays and then for pesticides applied via soil and 

### 16 seed treatment applications. 

### 17 Lastly, I will conclude with a comparison of 

### 18 our proposed decision-making process with other decision 

### 19 frameworks that we've identified in the literature. Before 

### 20 describing the frameworks themselves, I think it's important to 

### 21 discuss some of the attributes or factors that shape their 

### 22 development and, as a consequence, their overall scope. 

### 23 First of all, the agencies represented here 

### 24 are responsible for assessing ecological risk of over 1,000 

### 25 active ingredients for conventional pesticides. As a result, 

### 26 one attribute that shaped the proposed risk assessment process

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1 for bees was the need to efficiently screen out pesticides that 

### 2 posed little or no potential risk concern to bees from those 

### 3 that have a potential risk concern. 

### 4 To accomplish this, we are proposing a tiered 

### 5 process that begins with, at first, in Tier I, with a simple 

### 6 screening level assessment. And by design, this Tier I process 

### 7 is both conservative and quantitative in nature. And by that, I 

### 8 mean that it relies on high-end estimates of exposure and 

### 9 results in a derivation of a numeric risk quotient. 

### 10 The proposed Tier I assessment considers 

### 11 effects measured at the individual level of biological 

### 12 organization, under controlled laboratory conditions. Tiers II 

### 13 and III involve increasing information requirements that are 

### 14 typically satisfied by studies conducted in the field. And as a 

### 15 result, Tiers II and III involve a greater environmental realism 

### 16 in pesticide exposure compared to Tier I. 

### 17 In some cases, pesticide exposure information 

### 18 generated in Tiers II and III may be used to refine exposure 

### 19 estimated in Tier I. Importantly, however, in Tiers II and III, 

### 20 effects are evaluated at the colony level. 

### 21 Our proposed frameworks are also shaped by the 

### 22 need to rely largely on toxicity studies which have established 

### 23 regulatory guidelines; however, we and others have noted 

### 24 significant gaps in the exiting battery of toxicity test 

### 25 guidelines for bees. Therefore, these frameworks include 

### 26 placeholders for several additional studies for which test

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1 methods have not been fully vetted as regulatory guidelines, but 

### 2 protocols are available. 

### 3 The proposed decision frameworks also focus on 

### 4 major exposure pathways of concern to honey bees and are also 

### 5 shaped by the need to have sufficient information available from 

### 6 which a quantified pesticide exposure from these pathways. 

### 7 For example, some exposure pathways, such as 

### 8 ingestion of contaminated surface water are considered less 

### 9 important routes of exposure for honey bees, relative to direct 

### 10 contact and the consumption of pollen and nectar. 

### 11 Other pathways, such as consumption of plant 

### 12 guttation fluid may contain significant concentrations, but the 

### 13 degree to which honey bees rely on this source of water and 

### 14 nutrients has proven difficult to quantify. 

### 15 And again, later this morning, Ms. Christina 

### 16 Wendel will provide further evaluation of the selection of 

### 17 exposure pathways considered in this risk assessment process. 

### 18 The proposed decision frameworks also differ depending on the 

### 19 type of pesticide application which is consistent with the 

### 20 conceptual models that I presented previously. 

### 21 And lastly, decisions on moving from one tier 

### 22 to the next, which incorporate both risk assessment and risk 

### 23 management considerations are based on multiple lines of 

### 24 evidence and considerations of uncertainty. 

### 25 In addition to these attributes, I also wish 

### 26 to make several points regarding the potential application of

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1 these frameworks and future pesticide risk assessments involving 

### 2 honey bees. First, I'll be describing them as one moves from 

### 3 the top in Tier I, down to the bottom in Tiers III. 

### 4 As we envision this top to bottom application 

### 5 of the frameworks, may be more typical of their use and new 

### 6 pesticide risk assessment where the information base is just 

### 7 being developed. 

### 8 However, for existing pesticides, relevant 

### 9 information may be available at multiple tiers at the onset of a 

### 10 risk assessment, and in this case, the expectation would be that 

### 11 the decision-making process would reflect all available 

### 12 information available at the time of the assessment and may 

### 13 involve starting at various tiers simultaneously in the 

### 14 assessment. 

### 15 The second bullet emphasizes that the decision 

### 16 frameworks are intended to reflect an iterative process, whereby 

### 17 information gained at higher tiers can and should be used to 

### 18 refine information and assessments conducted at the lower tiers 

### 19 and vice-versa. 

### 20 Third, these decision frameworks are intended 

### 21 to be used as a guide to risk assessors and not as a rigid and 

### 22 inflexible recipe for decision-making. 

### 23 And then finally, while the frameworks reflect 

### 24 current state of the sciences as we understand it, the 

### 25 expectation is that as information and test guidelines are 

### 26 developed in the future, for example, for non-Apis bees,

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1 appropriate modifications will be made to the frameworks. 

### 2 So I will first begin by stepping through the 

### 3 proposed decision framework for assessing risk for honey bees to 

### 4 foliar spray application of pesticides, and this would include 

### 5 systemics as well as non-systemics. 

### 6 The first step in this process is to collect 

### 7 all relevant information regarding the nature of its pesticide 

### 8 and its use pattern, and this is a routine step in the problem 

### 9 formulation phase that includes gathering information on the 

### 10 pesticide's chemical, physical and toxicological properties, as 

### 11 well as detailed information on its environmental fate. 

### 12 Information on the timing and location and type and 

### 13 attractiveness of crops to which pesticides are being applied is 

### 14 also evaluated. 

### 15 Shown here, the Tier I begins with assembling 

### 16 this information and asking the question whether or not exposure 

### 17 of adults or brood to the pesticide is likely to be a potential 

### 18 concern. 

### 19 For outdoor spray applications, the answer 

### 20 would likely be yes, but for some other applications such as 

### 21 indoor or greenhouse uses, the answer would likely be no. 

### 22 In cases where pesticide exposure is not 

### 23 considered a potential concern at this stage, a presumption of 

### 24 minimal risk would be made. If exposure is considered to be a 

### 25 concern, then three major exposure routes would be evaluated in 

### 26 the Tier I part of the assessment. The first of these includes

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1 direct contact of adult foraging worker bees to the pesticide 

### 2 which intercept the spray droplets during flight. 

### 3 We have proposed a process to derive 

### 4 conservative or high-end estimated environmental concentration, 

### 5 abbreviated EEC, to the bees via contact exposure, as depicted 

### 6 in Box 3(a) of this diagram. 

### 7 The derivation of this EEC and other EECs used 

### 8 in Tier I will be discussed later by Ms. Kris Garber. As shown 

### 9 in Box 4(a), an acute risk quotient, or RQ, is then determined 

### 10 by the ratio of the contact EEC to the acute contact LD50. 

### 11 The second exposure route that would be 

### 12 evaluated is oral exposure of adult foraging bees. And again, 

### 13 here a conservative estimate of exposure via consumption and 

### 14 nectar is calculated as shown in Box 3(b) and an acute oral RQ 

### 15 is determined as a ratio of this oral EEC to the acute oral LD50 

### 16 for adult honey bees. And that's shown in Box 4(b). 

### 17 In addition, we propose that a measure of 

### 18 chronic risk to adult foragers be determined in Tier I by 

### 19 dividing the same EEC by the no-observed adverse effect 

### 20 concentration, or NOAEC, from a chronic toxicity study on adult 

### 21 bees. As noted with the asterisks, a regulatory guideline is 

### 22 not yet available for this study, although various protocols 

### 23 have been described in the literature. 

### 24 In the third exposure pathway proposed for 

### 25 evaluation in Tier I is oral exposure to developing honey bee 

### 26 brood. An oral EEC for larvae would be determined, as shown in

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1 Box 3(c) and the acute and chronic oral risk quotient values 

### 2 would be calculated using acute oral LD50 and a chronic oral 

### 3 NOAEC from the larval toxicity test. 

### 4 We note again that the regulatory guidelines 

### 5 have yet to be developed or finalized for measuring larval 

### 6 toxicity in the laboratory, and these would need to go through 

### 7 the formal guideline development process. 

### 8 Once these RQ values are determined, they will 

### 9 be compared to the proposed level of concern, abbreviated LOC. 

### 10 The proposed LOC for evaluating acute risk is 0.4, and that 

### 11 proposed for evaluating chronic risk is 1.0. The technical 

### 12 basis of these LOC values will be described later by Dr. 

### 13 Steeger. 

### 14 As the LOC comparisons are used to inform the 

### 15 decision to proceed with additional refinement or move on to 

### 16 Tier II; however, not explicitly depicted in Box 5, is that this 

### 17 decision would not only include a comparison to the LOC values, 

### 18 but also other lines of evidence, including available ecological 

### 19 incident reports, pesticide mode of action and uncertainty in 

### 20 the available information base. 

### 21 After considering the LOC, other lines of 

### 22 evidence, if no significant risk is indicated then a presumption 

### 23 of minimal risk would be made and documented. If risk could not 

### 24 be ruled out at this Tier I screening phase, then other 

### 25 information could be considered for refining the exposure 

### 26 estimate, which may include residue information derived from

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1 crop magnitude of residue studies which are routinely submitted 

### 2 to the Agency. Additional detail on this refinement step will 

### 3 be provided by Mr. Baris in a subsequent presentation. 

### 4 If the applicable and refined RQ value is 

### 5 recalculated using this refined information and other lines of 

### 6 evidence are considered, the decision then becomes, again, 

### 7 whether to presume minimal risk or continue with the risk 

### 8 assessment process. As shown in Box 8, the next step involves 

### 9 considering the available risk mitigation options and 

### 10 uncertainties in the assessment and other lines of evidence as 

### 11 part of this decision to move on to Tier II. 

### 12 The risk mitigation option, such as modifying 

### 13 the pesticide application rates, timing of application, types of 

### 14 crops being treated, could then be evaluated by proceeding back 

### 15 through the Tier I process, as indicated by this dashed black 

### 16 arrow on the right of the diagram. 

### 17 If after weighing this information, including 

### 18 a dialogue with risk managers, information from Tier II studies 

### 19 is determined to be necessary, then different types of studies 

### 20 could then be considered. 

### 21 One type of study may be an exposure-based 

### 22 field or semi-field residue study in which pesticide residues in 

### 23 pollen and nectar are quantified using pesticide applied to a 

### 24 treated crop. Information from this study could then be used to 

### 25 refine the estimates of pesticide exposure from pollen and 

### 26 nectar used earlier in Tier I.

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1 Another type of study would be considered in 

### 2 Tier II as a semi-field effects-based study, such as a tunnel 

### 3 enclosure or high feeding study. Semi-field studies provide 

### 4 pesticide effects information gathered at the whole hive level. 

### 5 And endpoints from these studies may include, but are not 

### 6 limited to worker and brood mortality, brood development, 

### 7 foraging activity, behavioral observation, colony strength and 

### 8 survival. Additional information on the Tier II semi-field 

### 9 studies will be provided by Mr. DeCant Later this afternoon. 

### 10 Once this information is gathered from the 

### 11 Tier II studies, other lines of evidence are considered as well 

### 12 as the uncertainties and potential risk to honey bees is again 

### 13 assessed. The outcome of this assessment may be a presumption 

### 14 of minimal risk, or if risk is identified, available risk 

### 15 mitigation options would be considered and evaluated, as 

### 16 indicated in Box 11. 

### 17 Although not shown here, evaluation of these 

### 18 mitigation options might also involve preceding back to the 

### 19 beginning of the process. After evaluating these risk 

### 20 mitigation option and considering, again, all the lines of 

### 21 evidence and uncertainties, a decision would be made to proceed 

### 22 to Tier III. 

### 23 And Tier III, as we've depicted it, includes 

### 24 full field studies that are designed to address specific 

### 25 uncertainties raised in the earlier tiers. An example of this 

### 26 might be the design and conduct of a field study specifically to

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1 track overwintering success of colonies following pesticide 

### 2 exposure. 

### 3 As shown in Box 13, the results of the Tier 

### 4 III field studies are then evaluated and available risk 

### 5 mitigation options are considered, and again, along with other 

### 6 lines of evidence and uncertainties. Risk associated with these 

### 7 mitigation options may be evaluated, as indicated by this black 

### 8 arrow, and risk is then characterized. 

### 9 This risk characterization may lead to a 

### 10 presumption of minimal risk or it may lead to a presumption of 

### 11 risk from the Tier III studies, as well as the other earlier 

### 12 information. Dr. Tom Steeger will be providing additional 

### 13 information on this risk characterization step this afternoon. 

### 14 The proposed decision framework for risk 

### 15 assessment of soil application and seed treatment application 

### 16 begins in a similar way that is depicted for foliar application. 

### 17 That is, information on the pesticide and the nature of its 

### 18 application are evaluated and questions are asked whether or not 

### 19 exposure is a potential concern to adults or brood. 

### 20 If there are little or no potential exposure 

### 21 of adults or brood, then a presumption of minimal risk is made. 

### 22 But if there is potential for exposure, then conservative 

### 23 estimates of oral exposure to adult worker bees and larvae are 

### 24 then determined, as indicated in Boxes 3(a) and 3(b) of this 

### 25 figure. 

### 26 As noted in Boxes 4(a) and 4(b), Tier I

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1 screening level, acute and chronic risk quotients are then 

### 2 calculated using these EECs and the relevant toxicity endpoints 

### 3 from the acute and chronic oral toxicity studies to adults and 

### 4 larvae are considered. 

### 5 As noted in the conceptual model and in the 

### 6 White Paper for seed treatments, exposure to bees from seed coat 

### 7 dust has been shown to be an important route of concern and the 

### 8 focus of numerous ecological incident reports. 

### 9 At this time, we're not proposing a risk 

### 10 assessment process for this exposure pathway, primarily because 

### 11 methods for predicting a priori pesticide exposure from drift of 

### 12 seed coat dust are not yet available for regulatory application 

### 13 and because the Agency is currently working to mitigate this 

### 14 exposure pathway based on modifications of seed treatment 

### 15 methods, seed planting equipment and associated practices. 

### 16 In addition, if there's a potential exposure 

### 17 for offsite drift of spray applications of soil, this would be 

### 18 addressed, as described previously, for the foliar spray 

### 19 applications after accounting for the fraction of pesticide that 

### 20 is subject to drift. 

### 21 As shown in Box 5, the acute and chronic risk 

### 22 quotients are then compared against the same LOC values as 

### 23 described previously. These RQ values are then considered along 

### 24 with other lines of evidence, such as incident reports, to 

### 25 determine whether a presumption of minimal risk can be made or 

### 26 if additional refinements are considered necessary in the Tier I

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1 exposure estimate. 

### 2 The remainder of the proposed decision 

### 3 framework is the same as I described for foliar sprays. 

### 4 However, the design of tests in Tiers II and III would reflect 

### 5 the specific pesticide application methods being used. And the 

### 6 EECs calculated in Tier I would also differ in their method. 

### 7 Lastly, we thought it would be useful to place 

### 8 the proposed decision framework I just presented in the context 

### 9 of two recently published risk assessment schemes for bees. 

### 10 The first is presented in a scientific opinion 

### 11 document developed by the European Food Safety Authority, or 

### 12 EFSA, earlier this year. And the second is a decision scheme 

### 13 that was published by the Society of an Environmental Toxicology 

### 14 and Chemistry, SETAC, in 2011, and based on an international 

### 15 Pellston-style Workshop. These efforts were very useful to us 

### 16 as we developed the White Paper. 

### 17 And as a result, we note a number of 

### 18 similarities among the three assessment schemes. First of all, 

### 19 as indicated in the first bullet, they all involve a tiered risk 

### 20 assessment process that begins with assessing effects at the 

### 21 individual level, based mostly on laboratory studies. This is 

### 22 then followed by effects which are assessed at the colony level 

### 23 at higher tiers. 

### 24 As indicated in the second bullet, they all 

### 25 contain risk assessment schemes that are tailored to different 

### 26 application methods; for example, sprays, soil and seed

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1 treatment. 

### 2 Third, exposure to pesticides from contact 

### 3 routes as well as oral exposure is prominent in all three 

### 4 schemes, although the EFSA scheme does include a conservative 

### 5 estimate of dust exposure from seed treatments as a fraction of 

### 6 the overall calculated application rate. It's not clear at this 

### 7 time which methods would then be used to refine this screening 

### 8 level estimate. 

### 9 Fourth, all schemes recognize the need to 

### 10 assess both acute and chronic toxicity early in the risk 

### 11 assessment process. I wish to point out some of the differences 

### 12 that we've noted as well. First, one example is that the 

### 13 triggers used to move from Tier I to higher tiers differ in 

### 14 their form. 

### 15 The process proposed in the White Paper bases 

### 16 the Tier I trigger value on the ratio of the estimated dose to 

### 17 the corresponding toxicity value. The approach recommended in 

### 18 the EFSA Scientific Opinion document, bases its trigger on the 

### 19 ratio of the application rate in grams per hectare to the 

### 20 toxicity value. 

### 21 As application rate is obviously related to 

### 22 pesticide doses, the two approaches are similar in concept, but 

### 23 they differ in the details. Another difference is that non-Apis 

### 24 bees are explicitly included in the risk assessment schemes 

### 25 recommended by the SETAC Pellston Workshop and EFSA. However, 

### 26 these schemes acknowledge that additional test guideline

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1 development is needed before they can be fully implemented for 

### 2 non-Apis bees. 

### 3 Lastly, the proposed approach in the White 

### 4 Paper recommends characterizing risk by considering the overall 

### 5 weight of evidence at higher tiers, which would include results 

### 6 from semi-field and field studies, but also would include other 

### 7 lines of evidence provided by suitable studies from the 

### 8 literature and incident reports. It appears to us that the risk 

### 9 conclusions at higher tiers from these other schemes are largely 

### 10 limited to the results from the applicable semi-field and field 

### 11 studies. 

### 12 I want to thank you very much for your 

### 13 attention. At this time, myself or the team would be pleased to 

### 14 address your questions on the overall risk assessment process, 

### 15 noting that detailed questions related to components of this 

### 16 process are probably best reserved for the specific presentation 

### 17 this afternoon. Thank you. 

### 18 **DR. DANIEL SCHLENK:**Any questions? Dr. 

### 19 Berenbaum. 

### 20 **DR. MAY BERENBAUM:**This may be best reserved 

### 21 for the afternoon, but just as a point of clarification, the 

### 22 acute and chronic toxicity testing at Tier I is described in 

### 23 discrete life stages. Is there any consideration of a way to 

### 24 incorporate life cycle testing rather than just individual 

### 25 discreet stage testing, sort of risk, kind of concept? 

### 26 **MR. KEITH SAPPINGTON:**In Tier I, we've only

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1 described, as you say, the larval and the adult stages. The 

### 2 larval toxicity test, as I understand, that we are evaluating is 

### 3 one that could continue out through emergence, but at this time, 

### 4 it's quite challenging to get acceptable responses in the 

### 5 controls from this. 

### 6 But I would believe with additional 

### 7 refinement, we would be able to perhaps capture that part of the 

### 8 development cycle. For capturing the entire life cycle, I 

### 9 believe those are reserved now in Tier II, where we would be 

### 10 looking at the hive level. 

### 11 **DR. DANIEL SCHLENK:**Dr. Potter. 

### 12 **DR. THOMAS POTTER:**Yes. Just a quick 

### 13 question about the LOC values that you proposed. Would those be 

### 14 bright line values in a sense that the value is fixed as .4 and 

### 15 you have a chemical that is say, .41? You know, what's the 

### 16 process there? 

### 17 **DR. THOMAS SEEGER:**When a compound or an RQ 

### 18 value exceeds an LOC, that information is relayed back to the 

### 19 risk manager. The decision to transition to higher tier levels 

### 20 or the degree to which the LOC is exceeded, that's a risk 

### 21 management decision. 

### 22 So would you consider it a bright line? It is 

### 23 a bright line in the sense that the LOC is fixed for screening 

### 24 level assessment. Whether it triggers the need to do higher 

### 25 tier assessments is a risk management decision. 

### 26 **DR. DANIEL SCHLENK:**Dr. Bradbury, you want

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1 to add a little bit more to that? 

### 2 **DR. STEVEN BRADBURY:**That LOC isn't provided 

### 3 in isolation from other information. So your .41 versus .39, 

### 4 what we want to know is what do we know about the mode of 

### 5 action; what does the dose response curve look like? 

### 6 All that information that Keith's been 

### 7 describing in the conceptual model would all come into play in 

### 8 terms of making a decision about what the uncertainties are and 

### 9 what would be the magnitude and the nature of the uncertainties 

### 10 around that threshold. 

### 11 But Dr. Steeger is right in that that LOC 

### 12 value is a value we use to make sure we pause, let's think, 

### 13 let's talk, let's look at all the information we got and make 

### 14 some decisions about what the next most logical step would be. 

### 15 **DR. DANIEL SCHLENK:**Okay. Dr. McManaman. 

### 16 17 **DR. JAMES MCMANAMAN:**Is it the Agency's view 

### 18 that the LD50 is the most conservative method estimate of 

### 19 toxicity for larvae or have you considered other possibilities? 

### 20 **DR. THOMAS STEEGER:**The LD50 and the LC50 are 

### 21 the traditional measures that the Agency relies on for screening 

### 22 level because it's considered the most precise estimate from the 

### 23 acute toxicity studies where the confidence limits around that 

### 24 estimate are at their narrowest point. 

### 25 **DR. JAMES MCMANAMAN:**But it may be the most 

### 26 reliable, but you view it as the most conservative estimate,

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1 especially for this first tier of screening? 

### 2 **DR. THOMAS STEEGER:**Whether it's the most 

### 3 conservative value, I mean, certainly, there are more 

### 4 conservative values that could be selected, but again, the 

### 5 precision that's surrounds those, the confidence limits that 

### 6 would be surrounding those estimates would be highly variable. 

### 7 At this point, that's the traditional measure that we, as a 

### 8 regulatory authority, as well as other regulatory authorities 

### 9 have used as a conservative measure of acute toxicity. 

### 10 **UNIDENTIFIED SPEAKER:**I wondered the same 

### 11 thing, actually. 

### 12 **DR. DANIEL SCHLENK:**Excuse me. Somebody 

### 13 on the telephone, can you please mute your line again, please? 

### 14 Thank you. 

### 15 **MR. KEITH SAPPINGTON:**I'd like to follow-up 

### 16 on that question. Again, I agree that the endpoint selected 

### 17 from the acute studies, such as the LC50 and the LD50 is the 

### 18 most precise estimate, but we then apply this level of concern 

### 19 to that. Dr. Steeger will be presenting the basis for that, so 

### 20 I don't want to get into that in significant detail. Part of 

### 21 the basis is the extrapolation down to a lower effect level, in 

### 22 this case, a ten percent effect level. 

### 23 **DR. DANIEL SCHLENK:**Dr. Bradbury. I saw 

### 24 your hand up. Did you want to -- 

### 25 **DR. STEVEN BRADBURY:**Keith beat me to the 

### 26 punch. The concept is a dose response relationship factors into

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1 that LOC. So it's a different way to get at the concept, but I 

### 2 think in the talks this afternoon it'll come out in more detail. 

### 3 **DR. DANIEL SCHLENK:**Dr. McManaman. 

### 4 **DR. JAMES MCMANAMAN:**I understand the 

### 5 concepts here, but whereas with adult bees I can see that it 

### 6 might be the appropriate measurement, but I'm concerned that for 

### 7 larvae, it may not be the appropriate measurements. So that was 

### 8 the basis of my question. 

### 9 **DR. DANIEL SCHLENK:**Okay. Dr. Pettis. 

### 10 **DR. JEFF PETTIS:**Just a point of 

### 11 clarification about, currently the White Paper is considered 

### 12 honey bees as a surrogate for all pollinators. And there's 

### 13 little consideration of the other life history for solitary 

### 14 bees; is that correct? 

### 15 **DR. THOMAS STEEGER:**You're correct that 

### 16 the proposed framework is using the honey bee as the focus and 

### 17 we are asking the SAP to comment on the extent to which it can 

### 18 be applied to non-Apis bees as well. But those measures of 

### 19 effect are based on individual bees which would presumably 

### 20 relate to solitary native bees. 

### 21 And at higher tier assessments, you're looking 

### 22 at colony level effects that would presumably speak to potential 

### 23 effects to social and non-Apis bees and as will be pointed out, 

### 24 and was already pointed out in preliminary presentations, but 

### 25 will be further discussed by Kris Garber, the measures of 

### 26 exposure are through diet and through contact which would

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1 presumably be relevant to ground nesting bees, solitary bees or 

### 2 non-Apis bees, just as it is relevant to the honey bee. 

### 3 **DR. DANIEL SCHLENK:**That was Dr. Steeger. 

### 4 Any other questions about the framework? 

### 5 (No response.) 

### 6 Okay. So now I think we're ready for Ms. 

### 7 Wendel, is that right? 

### 8 **MS. CHRISTINA WENDEL:**Hello. My name is 

### 9 Christina Wendel and I'm a biologist in the Environmental Fate 

### 10 and Effects Division of the Office of Pesticide Programs. And 

### 11 thank you for the opportunity to address the panel. 

### 12 This morning I will be talking about the 

### 13 exposure of honey bees to pesticides, the exposure routes that 

### 14 were considered and their overall relative importance in terms 

### 15 of major versus minor routes of exposure. 

### 16 Discussion of the potential exposure pathways 

### 17 is presented in the White Paper as part of the conceptual model 

### 18 in Section II, the Tier I exposure assessment in Section III, 

### 19 and the drinking water consumption rate analysis is in Appendix 

### 20 Number 2. This presentation relates to Charge Question Number 

### 21 7. 

### 22 I will be presenting the exposure routes 

### 23 identified and considered in the conceptual model as presented 

### 24 and described earlier by Mr. Sappington. I will also briefly 

### 25 discuss and describe the rationale used in determining the 

### 26 exposure routes identified for use in the Tier I exposure

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1 assessment, as well as those routes that were not included. 

### 2 Ms. Garber will follow this presentation with 

### 3 a more in-depth analysis of our proposed Tier I exposure 

### 4 assessment and the primary routes of exposure that were chosen 

### 5 and quantified. 

### 6 For the Tier I exposure assessments, three 

### 7 types of exposure routes were considered. These are contact, 

### 8 oral, and inhalation exposure. As identified previously, 

### 9 exposure routes differ with the type of application, which may 

### 10 include foliar sprays, soil applications, and seed treatments. 

### 11 In addition, exposure routes also differ based 

### 12 on the type of bee. Exposures differ for larval and adult bees 

### 13 and for the castes of the bee, which include workers, drones, 

### 14 and queen bees. 

### 15 The next few slides provide visual 

### 16 representations of exposure routes for bees based on both the 

### 17 application methodology and the type of bee. Again, this slide 

### 18 provides a visual representation of a honey bee and depicts the 

### 19 potential exposure routes for forager bees for foliar spray 

### 20 applications. 

### 21 For foliar spray applications, adult forager 

### 22 worker bees have the greatest potential for exposure as they 

### 23 leave the hive. Forager bees have direct contact with 

### 24 folairly-applied pesticides via direct spray and contact with 

### 25 pesticide residues on foliage and soil. 

### 26 Forager bees also are potentially exposed to

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1 pesticide residues via the collection of pollen, nectar, 

### 2 drinking water and plant resins or propolis that may be brought 

### 3 back to the hive. Another route of exposure for forager bees 

### 4 includes oral exposure through the diet from the consumption of 

### 5 contaminated pollen, nectar and/or drinking water. 

### 6 It is important to note that it was assumed in 

### 7 the Tier I exposure method that the major route of oral exposure 

### 8 is through the actual consumption of food. However, there could 

### 9 potentially be a dose of exposure for the forager bee from the 

### 10 carrying transport of pollen, nectar, and/or drinking water back 

### 11 to the hive. But we expect that the dose obtained from the 

### 12 consumption of food would be greater and productive of any dose 

### 13 obtained during the transport of pollen, nectar, and/or drinking 

### 14 water to the hive. 

### 15 Therefore, the oral exposure obtained from 

### 16 diet and not during the collecting phase was considered to be 

### 17 the oral route of exposure. During flight, forager bees may 

### 18 also be exposed via inhalation of spray droplets and/or the 

### 19 gassiest phase of volatilized chemicals. 

### 20 This slide provides a visual representation of 

### 21 the honey bee and depicts the potential exposure routes for hive 

### 22 bees for foliar spray applications. Adult bees living within 

### 23 the hive include workers, for example, nurse bees, as well as 

### 24 drones and queen bees. 

### 25 The adult hive bees are exposed primarily 

### 26 through the oral route of exposure from the consumption and

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1 processing of pollen and nectar into bee bread, honey, brood 

### 2 food and/or royal jelly, or the consumption of drinking water. 

### 3 There is no longer contact exposure to direct 

### 4 spray; however, contact exposure can occur within the hive 

### 5 through the transfer of pollen and/or nectar from forager bees 

### 6 to the hive worker bees, as well as direct contact with comb wax 

### 7 and propolis. 

### 8 Queen bees can also be exposed to direct spray 

### 9 and inhalation spray of spray droplets during mating and 

### 10 orientation flights; however, the dominate exposure route for 

### 11 queen bees is based on the exposure route as she consumes royal 

### 12 jelly throughout her life in the hive. 

### 13 For larval bees, items must be brought back to 

### 14 the hive and processed prior to larval exposure. Larval bees 

### 15 are exposed to the ingestion of the processed pollen and nectar, 

### 16 as royal jelly and/or brood food. As is the case for all adult 

### 17 and larval bees within the hive, inhalation of the gassiest 

### 18 phase of a volatilized chemical is also a potential exposure 

### 19 route. 

### 20 This slide also provides a visual 

### 21 representation of the honey bee and depicts the potential expose 

### 22 routes for forager and hive bees for soil and seed treatment 

### 23 applications. For soil and seed treatment applications, there 

### 24 is no longer contact exposure to direct spray. Instead, for 

### 25 seed treatment applications, there is potential contact exposure 

### 26 resulting from seed coat dust abrasion.

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1 In addition, for both adult and larval bees, 

### 2 oral exposure through the diet remains a major route of exposure 

### 3 for soil and seed treatment applications. For systemic 

### 4 compounds, translocation and route uptake occurs, resulting in a 

### 5 transfer of a chemical into the growing plant tissues and 

### 6 exudates, including pollen, nectar, as well as drinking water. 

### 7 Although the magnitude of exposure may be 

### 8 different between forager bees and hive bees, there is the 

### 9 potential for contaminates to be brought back to the hive and 

### 10 transferred to the bees living within the hive, based on 

### 11 grooming, the transfer of the collected items to the hive bees, 

### 12 the processing of pollen and/or nectar into bee bread, honey, 

### 13 brood food and/or royal jelly, the ingestion of the processed 

### 14 pollen and nectar products, as well as direct contact with comb 

### 15 wax and/or propolis. 

### 16 Again, for both adult and larval bees, 

### 17 inhalation is a potential exposure route in the hive from 

### 18 exposure to the gassiest phase of a volatilized chemical. 

### 19 During the Tier I exposure assessment, the 

### 20 exposure routes presented within the conceptual models for all 

### 21 application methodologies and chemical classes, including 

### 22 systemic and non-systemic compounds were examined and 

### 23 considered. 

### 24 Based on our review of the available 

### 25 information, the major routes of exposure of bees to pesticides 

### 26 are contact from direct spray for foliar spray application and

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1 oral exposure from the ingestion of the contaminated pollen and 

### 2 nectar for all pesticide application methods. 

### 3 The primary routes of exposure are shown here 

### 4 in green. These two routes of exposure are quantified in the 

### 5 Tier I exposure assessment by generating estimated environmental 

### 6 exposure concentrations, EECs. The process used to validate the 

### 7 Tier I exposure method will be discussed in the next 

### 8 presentation. 

### 9 For contact exposure, direct spray is the most 

### 10 conservative exposure route for forager bees and covers the 

### 11 other potential contact exposure routes shown here in blue, 

### 12 including contact with contaminated foliage, soil, comb wax, 

### 13 propolis and/or pollen. 

### 14 For oral exposure, the ingestion of 

### 15 contaminated pollen and/or nectar are assumed to be protective 

### 16 of other food consumed by bees, shown here in blue, and include 

### 17 honey, bee-bread, royal jelly, and brood food. 

### 18 Contact exposure via dust, oral exposure 

### 19 resulting from consumption of contaminated drinking water and 

### 20 inhalation exposure from spray droplets and/or the gassiest 

### 21 phase of a volatilized chemical, shown here in red, were 

### 22 considered and evaluated separately and will be discussed in the 

### 23 next several slides. 

### 24 Contact exposure from abraded dust as a result 

### 25 of seed treatment application is a potential major route of 

### 26 exposure to bees. Numerous incidents related to abraded dust

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1 from seed treatment applications have been reported in North 

### 2 America. As a result, the agencies are actively working with 

### 3 registrants, growers, applicators and seed equipment 

### 4 manufacturers to minimize this exposure pathway. Different 

### 5 technologies, stewardship programs and a variety of different 

### 6 mitigation measures, including label language, have been 

### 7 developed and research in these areas continues. 

### 8 Mitigation measures have been identified, 

### 9 including BMPs, new seed coat technologies, and alternative low 

### 10 dust lubricating agents to replace high dust producing 

### 11 lubricants like talc and graphite. In addition, best management 

### 12 practices are being considered and developed. A review of these 

### 13 processes will continue to occur as these technologies and 

### 14 practices advance over time. 

### 15 Although dust is considered to be a major 

### 16 exposure route, efforts are actively being made to minimize this 

### 17 route of exposure to bees. As a result, exposure to dust was 

### 18 not included in the Tier I screen for all pesticides. This 

### 19 issue is associated only with seed treatment applications and as 

### 20 a result, will be dealt with on a case-by-case basis for 

### 21 relevant chemical and application types. 

### 22 Water is collected for two primary reasons. 

### 23 First, to regulate the temperature within the hive through 

### 24 evaporative cooling to maintain conditions for healthy brood, 

### 25 and second, to use in preparing larval brood food by nurse bees. 

### 26

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1 Water is not stored within the hive; rather, 

### 2 it must be collected by foraging bees when needed by the colony. 

### 3 To reiterate, it is assumed in the Tier I Exposure Method that 

### 4 the major route of oral exposure is through the actual 

### 5 consumption of food, recognizing that there could potentially be 

### 6 a dose of exposure for the forager bee from the carrying and 

### 7 transport of the pollen nectar and/or drinking water back to the 

### 8 hive. But again, we expect that the dose obtained through the 

### 9 consumption of food would be greater and protective of any dose 

### 10 obtained through the transport of these materials. 

### 11 EPA completed an analysis to estimate 

### 12 potential exposure doses bees could receive from consuming 

### 13 contaminated drinking water. This analysis is presented in 

### 14 detail in Appendix 2 of the White Paper. Based on estimated 

### 15 pesticide concentrations in various sources of water potentially 

### 16 consumed by bees and water flux rate from the brown paper wasp, 

### 17 the results of this analysis indicate that if bees consume the 

### 18 majority of their water from ponds, including water adjacent to 

### 19 treated fields or puddles of water located on treated fields, 

### 20 the exposure is relative to dietary and direct spray are 

### 21 discountable. 

### 22 However, if bees drink a substantial amount of 

### 23 water from guttation fluid found in crops, or dew, the 

### 24 condensation on the plant, with exposure from dew occurring on 

### 25 crops folairly sprayed with pesticides, conservative exposures 

### 26 may be similar to or even exceed pesticide exposures through the

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1 diet or direct spray. Given the potential concern associated 

### 2 with exposure through drinking dew and guttation fluid, these 

### 3 exposure pathways were investigated further. 

### 4 Based on the results of EPA's analysis 

### 5 presented in Appendix 2 of the White Paper, pesticide exposures 

### 6 through dew and guttation fluid are not expected to be as 

### 7 significant when compared to the diet and direct spray because 

### 8 of two primary reasons. First, the relative importance of dew 

### 9 and guttation fluid as a source of drinking water to bees is 

### 10 unknown. In addition, dew and guttation fluid are expected to 

### 11 be present only during a portion of the day, typically in the 

### 12 morning, with a very conservative maximum of six hours overlap 

### 13 in foraging times of bees and the presence of dew and/or 

### 14 guttation fluid, preventing bees from drinking a substantial 

### 15 amount of water from these sources. 

### 16 Secondly, for many worker bees, pesticide 

### 17 doses, through the consumption of contaminated dew and/or 

### 18 guttation fluid, may be much smaller due to lower or 

### 19 non-existent drinking water consumption rates when compared to 

### 20 the percentage of water consumed from food. 

### 21 As shown in this slide, the estimated food 

### 22 consumption rates and resulting percentage of water consumed as 

### 23 part of food, including honey or nectar for adult worker and 

### 24 forager bees, ranges from 7 percent to greater than 100 percent 

### 25 of their daily water requirement. And again, these calculations 

### 26 are presented in Appendix 2 of the White Paper.

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1 Therefore, for many bees, pesticide exposure, 

### 2 through the consumption of either dew and/or guttation fluid is 

### 3 expected to be minimal because of higher amounts of water are 

### 4 likely consumed through food rather than from drinking water. 

### 5 Therefore, based on this analysis, pesticide exposures, through 

### 6 drinking water, was not included in the proposed Tier I exposure 

### 7 method for bees. 

### 8 Inhalation is a relevant exposure pathway for 

### 9 only highly volatile chemicals, including fumigants applied via 

### 10 foliar or soil application, where hives are in close proximity 

### 11 to the treated site. It is not a relevant exposure route for 

### 12 seed treatment applications. 

### 13 Based on a preliminary analysis conducted 

### 14 after the White Paper was developed, it was determined that 

### 15 exposure to bees via the inhalation route is substantially lower 

### 16 than the dietary exposure routes. Therefore, the inhalation of 

### 17 exposure pathway was not included as an exposure route in the 

### 18 proposed Tier I Exposure Method for bees. 

### 19 In summary, the primary routes of exposure 

### 20 that are quantified in the proposed Tier I method include 

### 21 contact resulting from direct spray and oral exposure from the 

### 22 dietary consumption of pollen and nectar. Although there are 

### 23 other potential exposure routes, these were evaluated and found 

### 24 to be minor in comparison to the direct contact and oral routes. 

### 25 26 Therefore, these other routes of exposure were

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1 not included within the proposed Tier I Exposure Assessment. We 

### 2 are seeking the input from the SAP on the proposed contact and 

### 3 oral exposure routes included in the Tier I Exposure Method, as 

### 4 well as those exposure routes that were not included in the Tier 

### 5 I Exposure Method for Bees. Thank you for time. The team and I 

### 6 can address any questions you may have. 

### 7 **DR. DANIEL SCHLENK:**Thank you. Any 

### 8 questions? Dr. James. 

### 9 **DR. ROSALIND JAMES:**Thank you. I thought the 

### 10 analysis you did was very good, but I have one question. You 

### 11 don't seem to consider Layer I, discussing about doing the 

### 12 bioassays for Tier I test, combining the oral and dermal 

### 13 together, whereas most bees are going to get both an oral 

### 14 exposure and the contact exposure. What's the reason for that? 

### 15 Or did I misunderstand that? 

### 16 **MS. KRISTINA GARBER:**You didn't misunderstand 

### 17 that. We're in the proposed process as it's depicted in the 

### 18 White Paper. We would just treat those two routes separately. 

### 19 Potentially, we could add risk quotients. 

### 20 So one approach could be to add risk quotients 

### 21 of the two pathways and then compare them to an LOC. But we are 

### 22 in the proposed process of treating them separately and the test 

### 23 would be conducted separately as well. 

### 24 **DR. THOMAS SEEGER:**I would like to add to 

### 25 Chris's comment. The in vitro assay that's been proposed for 

### 26 larval toxicity testing, those larvae are being exposed sort of

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1 as a combination of both oral and contact exposure. So in a way 

### 2 that concern is accounted for by that assay. 

### 3 **DR. DANIEL SCHLENK:**Okay. Dr. Tarpy. 

### 4 **DR. DAVID TARPY:**You had two main routes of 

### 5 water that bees would forage on, one is ponds or kind of runoff, 

### 6 and the other would be dew. Where would things like irrigation 

### 7 systems that are often used and bees use as an important source 

### 8 of water that are also conduits for pesticide treatment, where 

### 9 would that fall among those two routes of exposure? 

### 10 **MS. KRISTINA GARBER:**I think we weren't 

### 11 actually accounting for the irrigation or even direct 

### 12 applications to irrigation canals. That really doesn't fit into 

### 13 the concept of the pond. If it's a direct application or a 

### 14 chemigation application, perhaps, if that's what -- okay. 

### 15 You're nodding "yes." 

### 16 I think that, you know, one way to deal with 

### 17 that could potentially be to take the labeled application rate 

### 18 as it appears, you know, for the chemigation application and use 

### 19 the drinking water ingestion rates that we've got in the paper 

### 20 and see if the assumptions that we've got presented here still 

### 21 hold. And if so, we could proceed as usual. But this kind of 

### 22 speaks to what Keith was talking about earlier about how our 

### 23 conceptual diagrams that we're presenting are kind of based on a 

### 24 suite of application methods that are pretty typical of 

### 25 agriculture. 

### 26 In this case, I mean, chemigation is a typical

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1 application as well. We could modify that conceptual model to 

### 2 account for this other scenario. 

### 3 **DR. DANIEL SCHLENK:**Dr. Pistorius. 

### 4 **DR. JENS PISTORIUS:**I have a question. You 

### 5 said the effective quantitative screening tool is not currently 

### 6 available for dust. If such methods are available, could you 

### 7 imagine incorporating those in the risk assessment scheme? I 

### 8 hope this is appropriate to ask, how long do you think it would 

### 9 take? 

### 10 The second is it is a technical issue also. 

### 11 So it is a question of risk assessment and risk management. If 

### 12 such risk management measures would be possible, do you think it 

### 13 is appropriate to take it out from the risk assessment 

### 14 completely? 

### 15 **DR. THOMAS STEEGER:**The issue of dust, 

### 16 abraded dust from treated seed coat, as we understand it, 

### 17 requires a number of factors. And the difficulty in quantifying 

### 18 the level of dust that could be generated that would allow us to 

### 19 predict it on a regular basis, we don't know if at this time 

### 20 such a method of prediction is possible. 

### 21 If one were developed, then yes, I think we 

### 22 would consider it, but at this point we're hoping that through 

### 23 best management practices, the likelihood of abraded dust can be 

### 24 reduced such that it won't represent major route of exposure. 

### 25 But clearly there are incidents that suggest -- as was evidenced 

### 26 in Germany in 2008 -- that abraded dust can serve as a source of

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1 exposure, given an alignment of factors that influenced the 

### 2 generation and movement of those dust from treated seed. 

### 3 The issue of management, as Mr. Sappington 

### 4 portrayed throughout the movement of the framework, risk 

### 5 assessment or risk management options are considered throughout 

### 6 the process. And if we can discount potential exposure through 

### 7 mitigation, that would be factored into the assessment. 

### 8 That's why the importance of maintaining close 

### 9 communication with the risk manager from the very start of the 

### 10 assessment process would enable us to better account for 

### 11 mitigation options that might be considered. 

### 12 **DR. DANIEL SCHLENK:**Do you have a follow 

### 13 up with that Mr. Pistorius? 

### 14 **MR. JENS PISTORIUS:**Just a second question. 

### 15 With the risk mitigation measures, this would also maybe be 

### 16 possible to imply, for instance, if there's a risk for close set 

### 17 up of hives near a field where there is guttation very likely. 

### 18 Would it be possible to make a link between risk assessment and 

### 19 risk mitigation measures there and apply with mitigation 

### 20 measures? 

### 21 **DR. THOMAS STEEGER:**The regulating pesticides 

### 22 at a nation level to account for the position of colonies near 

### 23 crops that may be producing guttation fluid, we still -- and we 

### 24 welcome panel input on this -- we're still uncertain as to the 

### 25 degree that bees would be attracted to plants when guttation 

### 26 fluids are actually being expressed from plants.

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1 If we had a better understanding of that route 

### 2 of exposure, I think we would be in a better position to 

### 3 consider whether it should be included as a routine route of 

### 4 exposure and whether mitigation could be consistently applied to 

### 5 that as a way of resolving it. 

### 6 **DR. DANIEL SCHLENK:**Dr. Potter. 

### 7 **DR. THOMAS POTTER:**I have two questions, if I 

### 8 may. The first has to do with the -- it's a question and a 

### 9 comment and it has to do with the drinking water exposure. 

### 10 In reading through Appendix 2, I was struck by 

### 11 what appears to be a considerable uncertainty in terms of the 

### 12 quantity of water that a bee may consume during the day. I 

### 13 looked at your assessment; perhaps that's all the information 

### 14 that's available. Not being a bee expert, I can't comment on 

### 15 that one way or the other. 

### 16 I will note, in that context of drinking water 

### 17 exposure that I looked at your calculations with regard to 

### 18 concentrations of pesticides in puddles. I think there was an 

### 19 error in units in application of the formula that you used, that 

### 20 you need to take a careful look at. 

### 21 My question connected to that is in looking at 

### 22 the possibility of pesticide contamination of infield puddles, 

### 23 did you look back at the work that was done in establishing the 

### 24 ECO Framework? It was about 10 years ago that ECOFRAM was on 

### 25 the scene. 

### 26 Buried in there, and apparently the

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1 information is still available on the website, there is a very 

### 2 nice little model that was developed to actually look at the 

### 3 possibility of residues accumulating in puddles infield. I 

### 4 think that that might be something that you'd want to take a 

### 5 look at. 

### 6 So my question for you is, did you look at 

### 7 that ECO Framework at all in the context of your puddle 

### 8 assessment? 

### 9 **MS. KRISTINA GARBER:**No. We didn't look at 

### 10 the ECOFRAM report specifically for this method, but we can 

### 11 certainly do that. Thank you. 

### 12 **DR. THOMAS POTTER:**My second question is with 

### 13 regard to dust exposure, did you consider any other dust in the 

### 14 bees' environment that may contain pesticide residues? And this 

### 15 could include windblown soil, dust that is generated from soil 

### 16 or sprays that have been applied. It could indeed include a 

### 17 dust application or formulation that contains the active 

### 18 ingredient. 

### 19 My question for you is, what other forms of 

### 20 dust in the environment were looked at other than the 

### 21 descriptions you made about dust that may have been generated 

### 22 from abraded seed? 

### 23 **DR. THOMAS STEEGER:**The dust that was 

### 24 considered was that that was abraded from treated seed because 

### 25 it would be the most concentrated form of dust where the active 

### 26 ingredient would be present. So the dust that's potentially

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1 contributed through the movement of seeding equipment, 

### 2 disrupting soil or any type of tillage was not considered in our 

### 3 assessment. Again, the concentration, that would be considered 

### 4 from dust as a result of abraded seed coat would expected to be 

### 5 higher. 

### 6 **DR. THOMAS POTTER:**I agree it would be 

### 7 higher, but there may be some exposure there. We'll get into 

### 8 that, I think, when we talk about some of the questions. At 

### 9 least from my rationale, I think it might be worth taking a look 

### 10 at. 

### 11 **DR. DANIEL SCHLENK:**Any other questions? 

### 12 (No response.) 

### 13 Okay. According to my schedule and agenda, we 

### 14 have a lunch break scheduled. So let's go ahead and do that and 

### 15 be back at 12:45 and we'll resume at that time. Thanks. 

### 16 (Whereupon, at 11:41 a.m., a 

### 17 luncheon recess was taken.) 

### 18 * * * * * 

### 19 A F T E R N O O N S E S S I O N 

### 20 (12:50 p.m.) 

### 21 **DR. DANIEL SCHLENK:**Now that our permanent 

### 22 panel members are here, we can kind of go forward. Let's go 

### 23 ahead and get started. 

### 24 Before we get started, I believe you have some 

### 25 answers to Dr. Potter's question. 

### 26 **MS. KRISTINA GARBER:**Thank you. Dr. Potter,

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1 I did a little research on my lunch break and I just wanted to 

### 2 get back to you on the puddle comments that you had made. 

### 3 So first off, you had something about a unit 

### 4 error. And I was just hoping that you could note that in your 

### 5 written comments so that we can make sure we go back and take a 

### 6 look at that. 

### 7 **DR. THOMAS POTTER:**I absolutely intend to 

### 8 comment on that. I just wanted to point that out, you know, I 

### 9 went back and ran the numbers and got something different. So I 

### 10 will certainly put it in the record. 

### 11 **MS. KRISTINA GARBER:**Thank you. And then the 

### 12 other thing related to the ECOFRAM approach, it's my 

### 13 understanding that that's kind of based on a modification of the 

### 14 prism model. So we did actually consider that in the 

### 15 development of the model that we presented for estimating 

### 16 pesticide concentrations in puddles. 

### 17 And we chose to go with the model that we did 

### 18 just simply because it was a more simple approach, you know, 

### 19 using the simple equilibrium-based approach, it indicated that 

### 20 concentrations in puddle weren't a problem. So we didn't see 

### 21 the need to move further to a more refined approach. That's 

### 22 basically why we chose to proceed the way that we did. 

### 23 **DR. DANIEL SCHLENK:**Thanks. Okay. I 

### 24 guess you're up, Kristina. So go ahead and lead us on our next 

### 25 presentation, please. 

### 26 **MS. KRISTINA GARBER:**Great. Thank you, Dr.

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1 Schlenk. Thank you, members of the panel for allowing me to 

### 2 present on the Tier I Method for Estimating Exposure of Honey 

### 3 Bees to Pesticides. As a formal introduction, I'm senior 

### 4 biologist in the Environmental Fate and Effects Division of the 

### 5 Office of Pesticide Programs. 

### 6 Now, the material that I'm going to cover 

### 7 today is included in Section III in Appendix 1 of the White 

### 8 Paper. This material is relevant to Charge Questions 3, 4, 5, 

### 9 and 6. 

### 10 In this presentation, I'll give a brief 

### 11 overview of the Tier I Exposure Assessment Methodology, 

### 12 including a description of the purpose of a Tier I assessment, 

### 13 as well as a discussion of where this method fits into the 

### 14 decision framework that was presented earlier by Keith 

### 15 Sappington. 

### 16 As Christina Wendel discussed earlier, dietary 

### 17 exposure is a major exposure route of pesticides for honey bees. 

### 18 So in order to estimate pesticide exposures through the diet, 

### 19 it's necessary to understand what honey bees eat and how much. 

### 20 In this presentation I'll describe the 

### 21 proposed food consumption rates for worker larvae and for 

### 22 adults. I'll also discuss how conservative these consumption 

### 23 rates are when compared to other honey bees, as well as other 

### 24 species of bees. 

### 25 I'll then move on to describe the proposed 

### 26 method for estimating pesticide exposure from contact that

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1 results from foliar applications as well as the methods for 

### 2 estimating pesticide concentrations in pollen and nectar, 

### 3 following applications that are made via foliar spray, seed 

### 4 treatment and soil applications. 

### 5 These concentrations in pollen and nectar are 

### 6 used in combination with the proposed food consumption rates for 

### 7 adults and for larval worker bees in order to estimate doses 

### 8 that are received by these two groups of bees. When I'm saying 

### 9 "bees" here, I mean honey bees. 

### 10 Now I'll give a brief overview of the Tier I 

### 11 Exposure Assessment. Now, the goal of the Tier I Method is to 

### 12 generate reasonably conservative estimates of pesticide 

### 13 exposures to bees. This method is intended to estimate upper 

### 14 bound concentrations that are comparable to an upper 95th 

### 15 percentile exposure for honey bees. 

### 16 When evaluating whether a method generates 

### 17 reasonably conservative estimates of exposure, estimates should 

### 18 preferably be higher and generally be within one or two orders 

### 19 of magnitude of available exposure data, empirical exposure 

### 20 data. 

### 21 The Tier I Exposure Assessment Method is 

### 22 designed to allow risk assessors to confidently distinguish 

### 23 between those pesticides that do not pose a risk to honey bees 

### 24 and those that require further consideration. In this approach, 

### 25 we attempt to minimize Type II errors, which would involve 

### 26 concluding that there is no risk when there actually is a

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1 potential for effects. And at the Tier I level, Type I errors 

### 2 are more acceptable because additional refinements can always be 

### 3 made at higher tiers of the risk assessment. 

### 4 Now, this figure is a subset of the decision 

### 5 framework for foliar spray applications that was presented 

### 6 earlier today by Keith Sappington. And the focus of this 

### 7 particular slide is on the Tier I Exposure Assessment Method. 

### 8 As depicted in Boxes 1 and 2 of this figure, 

### 9 once the use information for a pesticide product is determined, 

### 10 the risk assessor would determine whether or not there's a 

### 11 potential exposure for adults or for brood to the pesticide. In 

### 12 making this determination, the risk assessor would need to 

### 13 consider various factors, such as the potential overlap of the 

### 14 pesticide application and the presence of honey bees on the 

### 15 field, the attractiveness of the crop to honey bees or plants 

### 16 that may be present in areas adjacent to the field, such as 

### 17 dandelions. 

### 18 If there is the potential for exposure, the 

### 19 risk assessor moves on to boxes 3 a, b, and c, which is the 

### 20 focus of this particular method. These involve estimation of 

### 21 exposures to adults and to larvae. These three boxes are the 

### 22 focus of the Tier I Exposure Assessment Methodology. For foliar 

### 23 spray applications, exposures are assessed separately for 

### 24 contact and for diet. 

### 25 As depicted in boxes 4 a, b, and c, estimated 

### 26 exposure values should be matched to their respective toxicity

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1 endpoints from contact and oral tests for adult honey bees and 

### 2 larvae to derive risk quotients. 

### 3 Tier I toxicity endpoints are expressed on a 

### 4 dose basis in units of micrograms active ingredient, or AI, per 

### 5 bee. Therefore, all exposure values are expressed on a dose 

### 6 basis. Boxes 4 b and c indicate that both acute and chronic 

### 7 risk quotients are calculated in the Tier I risk assessment. 

### 8 Both acute and chronic exposure estimates are represented by the 

### 9 highest single days' dose. 

### 10 Although a time-weighted estimated exposure 

### 11 value may be more representative of the exposure period in a 

### 12 chronic toxicity test, exposure occurring over a single day 

### 13 could potentially be sufficient to elicit effects. Therefore, 

### 14 in the Tier I approach, chronic exposure is conservatively 

### 15 represented by the highest single day estimated exposure. 

### 16 This figure is a subset of the decision 

### 17 framework for seed and soil treatments, again, with a focus on 

### 18 the Tier I Exposure Assessment Method. I'm not going to go into 

### 19 detail on this figure because it's very similar to the one I 

### 20 just discussed for foliar applications. But the one notable 

### 21 difference here is that there's no contact exposure that's 

### 22 included in this decision tree. So the major exposure route 

### 23 incorporated into the Tier I Exposure Assessment Method for soil 

### 24 and seed treatments is through the diet. 

### 25 Now I'll move on to discuss the honey bee food 

### 26 consumption analysis. And this is covered in detail in Appendix

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1 1 of the White Paper. The basis of the honey bee diet is pollen 

### 2 and nectar. Honey bees will consume pollen and nectar directly 

### 3 or they'll eat bee bread and honey, which are forms of pollen 

### 4 and nectar that are processed for storage in a hive. 

### 5 In addition, honey bees consume royal jelly 

### 6 and brood food, which are composed primarily of glandular 

### 7 secretions that are produced by nurse bees. The amount of food 

### 8 and specific consumption of a honey bees' diet varies by caste 

### 9 and by age. The diet of a worker bee changes over the course of 

### 10 its life to meet the energy requirements of its task which 

### 11 change with age. 

### 12 For instance, nurse bees consume the most 

### 13 pollen, compared to other worker bees, so that they can produce 

### 14 jelly to feed larvae and the queen. Foraging bees consume the 

### 15 most honey and nectar in order to meet the energy demands of 

### 16 flying. 

### 17 Because the proposed risk assessment method 

### 18 involves conducting separate risk assessments for brood and for 

### 19 adults at the Tier I level, separate food consumption rates for 

### 20 larvae and for adults are necessary to calculate the different 

### 21 pesticide exposures through the diet. 

### 22 The proposed food consumption rates for the 

### 23 Tier I Exposure Assessment are 292 milligrams of food per day 

### 24 for adults and 120 milligrams of food per day for larvae. As I 

### 25 indicated earlier, the Tier I Exposure Assessment is intended to 

### 26 produce reasonably conservative estimates of exposure.

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1 Therefore, the proposed food consumption rates 

### 2 are based on portions of the adult and larval life stages where 

### 3 honey bees are expected to be consuming the greatest amounts of 

### 4 food, and would therefore be expected to receive the highest 

### 5 pesticide exposures through the diet, when compared to honey 

### 6 bees at different ages. 

### 7 So for adults, the nectar forager bees consume 

### 8 the most food and for larvae, it's individuals that are aged 

### 9 five days. There are several assumptions related to the 

### 10 proposed food consumption rates. In this approach, it is 

### 11 assumed that exposures through consumption of nectar and pollen 

### 12 are conservative representations of potential exposures through 

### 13 consumption of honey and bee bread. 

### 14 This approach is likely to be conservative 

### 15 because it assumes that pesticides contained in honey and bee 

### 16 bread do not degrade while these foods are stored in the hive. 

### 17 For honey bees that consume honey, it's 

### 18 assumed that the estimated pesticide exposures can be related 

### 19 back to the original concentration of the pesticide and nectar. 

### 20 This is accomplished by accounting for the amount of sugar 

### 21 that's consumed by bees and the amounts of sugar that are 

### 22 present in honey and in nectar. This is necessary since 

### 23 pesticide concentrations are estimated in nectar of treated 

### 24 crops. 

### 25 Using this approach, the pesticide dose 

### 26 received by a honey bee consuming nectar would be a equivalent

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1 to a dose received by a honey bee consuming honey from the same 

### 2 source of nectar. It's also assumed that pollen and nectar 

### 3 consumption rates and resulting exposures are protective of 

### 4 exposures of honey bees, the pesticides through consumption of 

### 5 royal jelly and brood food. This is based on results from two 

### 6 studies that have observed concentrations of pesticides in royal 

### 7 jelly that were 100 times less than the food that was consumed 

### 8 by the nurse bees that produce that jelly. 

### 9 Finally, for the Tier I Exposure Assessment 

### 10 Method, we're proposing to use the estimated pesticide 

### 11 concentrations in or on crop foliage as a surrogate for 

### 12 pesticide concentrations in nectar and in pollen. 

### 13 So essentially, the pesticide concentration in 

### 14 nectar is treated in the Tier I method as being equivalent to 

### 15 the concentration in pollen. Therefore, for the Tier I Exposure 

### 16 Assessment Method, the proposed food consumption rates are based 

### 17 on consumption of pollen plus nectar. So I'll express that as 

### 18 total food consumption. 

### 19 In order to indentify the group of adult 

### 20 worker bees that have the highest food consumption rates, pollen 

### 21 and nectar consumption information were identified in the 

### 22 scientific literature. Now, this table provides food 

### 23 consumption rates for different adult worker bees, as identified 

### 24 by their task. 

### 25 The age range associated with each task is 

### 26 intended to be general and basically reflects the task that a

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1 worker will carry out at different ages of her life, with 

### 2 in-hive tasks being carried out by younger bees and foraging 

### 3 tasks that occur outside of the hive being carried out by older 

### 4 worker bees. 

### 5 Pollen consumption rates of worker honey bees 

### 6 are based on direct empirical measures published by Crailsheim, 

### 7 et al. in 1992. Nectar consumption rates are based on a 

### 8 commonly cited body of work published by Rortais, et al. in 

### 9 2005. 

### 10 These authors estimated sugar consumption 

### 11 rates of different types of worker honey bees based on energy 

### 12 requirement that were associated with the bees' specific tasks. 

### 13 The sugar consumption rates published by Rortais, et al. are 

### 14 converted here to nectar equivalence by accounting for the 

### 15 average sugar content of nectar, which is 30 percent. As you 

### 16 can see by this table, pollen consumption rates vary among bees, 

### 17 but are at least one order of magnitude below nectar consumption 

### 18 rates of the same bees. 

### 19 When we compare the total daily food 

### 20 consumption rates among the different worker bees, the nectar 

### 21 foraging bees had the highest food consumption rates. As you 

### 22 can see by this table, the majority of food consumption of this 

### 23 group of bees is represented by nectar, since the pollen 

### 24 consumption rate is four orders of magnitude lower. 

### 25 Since the food consumption rate has a large 

### 26 influence on the final estimated dose received by honey bees and

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1 for the nectar forager, this food consumption rate is 

### 2 represented by nectar consumption. We did a detailed review of 

### 3 the analysis conducted by Rortais, et al. for the nectar forager 

### 4 bee. 

### 5 According to the method published by Rortais, 

### 6 et al., the nectar consumption rate of the nectar foraging bee, 

### 7 as estimated, using five variables, including the sugar required 

### 8 for flying, the amount of sugar that's required for flying. The 

### 9 number of foraging trips that are made in one day, the duration 

### 10 of a single foraging trip, the fraction of time spent flying 

### 11 during a trip, and the amount of sugar present in nectar. 

### 12 One limitation to the approach is that there's 

### 13 a great deal of variability in these parameters. Also, the 

### 14 approach presented by Rortais, et al. did not account for the 

### 15 sugar requirements of bees while at rest. 

### 16 Because there's so much variability within 

### 17 each of these parameters, we conducted a distributional analysis 

### 18 using a Monte Carlo simulation that simultaneously accounted for 

### 19 variability in all five parameters. We also accounted for the 

### 20 resting metabolic requirements of nectar foragers. 

### 21 The results of the Monte Carlo simulation are 

### 22 depicted in this figure. The proposed food consumption rate for 

### 23 adult worker bees in 292 milligrams of food per day. This value 

### 24 is based on the median estimate of nectar consumption of nectar 

### 25 foraging bees in the simulation that was conducted. 

### 26 The median value is selected for two primary

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1 reasons. First, the median is considered most likely to 

### 2 represent food consumption of this group of bees. Second, use 

### 3 of the median avoids compounding conservative assumptions in the 

### 4 overall Tier I Exposure Method. 

### 5 The proposed food consumption value already 

### 6 targets the group of bees with the highest food consumption 

### 7 rates compared to other adult bees. As I indicated earlier, the 

### 8 goal of the Tier I Exposure Assessment Method is to estimate 

### 9 reasonably conservative exposures to bees. 

### 10 Now, the target of the reasonably conservative 

### 11 estimate is approximately the upper 95th percentile, and also 

### 12 I'll discuss later, the estimated pesticide concentrations in 

### 13 pollen and nectar represent an upper bound. 

### 14 If an upper bound food consumption rate for 

### 15 nectar foragers was also proposed, the final dietary dose would 

### 16 most likely exceed the desired 95th percentile for worker bees 

### 17 due to a combination of a upper bound estimate of residues and 

### 18 food with an upper bound estimate of food consumption. 

### 19 Next, I'd like to discuss the relative 

### 20 conservativeness of the proposed food consumption rate for adult 

### 21 bees when compared to other worker honey bees and drones, as 

### 22 well as other species of bees. This figure depicts the nectar 

### 23 and pollen consumption rates of different adult worker honey 

### 24 bees and drones. 

### 25 As you can see by this figure, the worker bee 

### 26 that forages for nectar has the highest food consumption rate

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1 among adult worker bees and drones, even when considering nectar 

### 2 and pollen consumption rates. 

### 3 Nectar foraging bees are obviously summer 

### 4 bees. Winter bees that maintain the functions of the hive have 

### 5 different energy requirements. The nectar forager food 

### 6 consumption rate is higher than the food consumption rate of 

### 7 winter workers, indicating that it is also conservative for 

### 8 winter bees. 

### 9 These data are presented in Appendix 1 of the 

### 10 White Paper. In some cases, ranges of food consumption rates 

### 11 were presented in this appendix. In this figure, the midpoints 

### 12 of those ranges are presented. This is worth noting here 

### 13 because the high-end estimate of nectar consumption of flying 

### 14 adult drones actually exceeds the proposed nectar consumption 

### 15 rate of nectar foraging workers; however, the low-end of the 

### 16 drone's nectar consumption rate is below the proposed value. 

### 17 Therefore, the proposed food consumption rate 

### 18 for the nectar forager is still considered to be representative 

### 19 for drones. One limitation to this analysis is that data were 

### 20 not located to estimate the food consumption rates of adult 

### 21 queens. And this represents a data gap. 

### 22 However, given that queens primarily consume 

### 23 royal jelly and that available data suggests that this food will 

### 24 contain at least 100 times lower pesticide concentrations, 

### 25 compared to the food that's consumed by nurse bees, the food 

### 26 consumption rate of queens would have to be at least 100 times

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1 greater than the proposed food consumption rate for the nectar 

### 2 forager in order for the queen to receive a higher pesticide 

### 3 dose through the diet. Although the queen likely consumes more 

### 4 food than other honey bees, it seems unlikely that she would 

### 5 consume two orders of magnitude more food. 

### 6 In summary, it appears that the proposed food 

### 7 consumption rate is conservative for other adult worker honey 

### 8 bees, as well as drones and queens. Before moving on, it's 

### 9 worth noting here that this talk is focused on the food 

### 10 consumption rates for the Tier I Exposure Assessment. In some 

### 11 cases, the risk assessor may choose to use different food 

### 12 consumption rates than the proposed value. 

### 13 For instance, if chemical-specific pesticide 

### 14 concentration data are available for pollen and nectar, the risk 

### 15 assessor may want to use the pollen and nectar consumption rates 

### 16 separately. In addition, the food consumption rates that are 

### 17 depicted here for different worker bees could also be used for 

### 18 colony level modeling. 

### 19 As I noted previously, honey bees also consume 

### 20 honey and that we're calculating food consumption rates on a 

### 21 nectar-equivalent basis. When we consider the relative food 

### 22 consumption rates among worker bees, when honey and pollen 

### 23 consumption is considered, the nectar forager is still the most 

### 24 conservative among adult bees. 

### 25 We also compared the proposed food consumption 

### 26 rate for adult worker honey bees to available food consumption

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1 rates for three other species of bees, including the bumblebee, 

### 2 the European mason bee and the alfalfa leaf-cutting bee. 

### 3 These data suggest that the proposed food 

### 4 consumption rate for adult honey bee workers is similar to that 

### 5 of the adult bumblebee and is greater than that of the European 

### 6 mason bee and the alfalfa leaf-cutting bees. 

### 7 This information suggests that the proposed 

### 8 food consumption rate for the Tier I Exposure Assessment for 

### 9 adult honey bees would be representative and somewhat protective 

### 10 for adults of these three non-Apis species. Please note that 

### 11 this information is discussed in Section 5.3 of the White Paper 

### 12 if you'd like any additional details. 

### 13 As with adult worker bees, the intention of 

### 14 the food consumption rate for honey bee larvae is to focus on 

### 15 the age of larvae that consume the most food compared to other 

### 16 ages. Over the course of the five-day unkempt period, the diet 

### 17 of larvae changes. During the first three days they eat brood 

### 18 food and royal jelly. During the fourth and fifth days, they 

### 19 eat honey and pollen. 

### 20 Worker larvae grow at an exponential rate over 

### 21 the course of their five-day developmental period. It's assumed 

### 22 that their food consumption rate doubles each day in order to 

### 23 meet the metabolic requirements of growth. The food consumption 

### 24 rates of larvae are based on information cited by Rortais, et 

### 25 al. that indicated that larvae consumed 120 milligrams of food 

### 26 over the course of the fourth and fifth days.

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1 The pollen consumption rates are based on 

### 2 empirical measures of pollen contained in the guts of larvae. 

### 3 It is assumed that the remainder of the 120 milligrams of food 

### 4 consumed over days four and five is honey that is diluted with 

### 5 sugar -- with water, to 45 percent sugar. Based on the 

### 6 assumption that the food consumption rate of larvae doubles 

### 7 every day, the food consumption rates were calculated as 

### 8 depicted in this table. 

### 9 The proposed food consumption rate of 120 

### 10 milligrams of food per day is based on the food consumption of 

### 11 the larvae at Day 5. This value represents consumption of 2.7 

### 12 milligrams of pollen per day and 117 milligrams of nectar per 

### 13 day. In this case, the amount of honey consumed is converted to 

### 14 nectar equivalence, assuming that the sugar content of nectar is 

### 15 30 percent and the sugar content of honey, which is diluted with 

### 16 water, is 45 percent. 

### 17 Next, I'd like to discuss the relative 

### 18 conservativeness of the proposed food consumption rate for 

### 19 worker larvae when compared to other castes of honey bees and 

### 20 then to other species of bees. This figure depicts the total 

### 21 food consumption rates of worker larvae at different days of 

### 22 this life stage. 

### 23 Please note that at on the X-axis, the caste 

### 24 of the larvae is provided along with the day of development in 

### 25 parentheses. The Y-axis represents food consumption rates where 

### 26 food is represented by royal jelly or brood food, honey, and

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1 pollen. 

### 2 This figure includes the food consumption 

### 3 rates for drone larvae at days 5 and 6. The food consumption 

### 4 rate for the five-day-old worker larvae appears to be consistent 

### 5 with that of the drone larvae. It should be noted here that the 

### 6 pollen consumption rates of drone larvae are unknown. In this 

### 7 figure, it's assumed that they are equal to the pollen 

### 8 consumption rates of workers at day 4 and 5. 

### 9 Given that pollen consumption rates are 

### 10 generally much lower than honey consumption rates, this 

### 11 assumption is unlikely to impact the conclusion that the 

### 12 five-day worker larval consumption rate of food is consistent 

### 13 with that of drone larvae. 

### 14 This figure includes the food consumption 

### 15 rates for queen larvae at days 1 through 5. These food 

### 16 consumption rates are calculated by assuming that queen larvae 

### 17 consumed 2.5 times the amount of food that's consumed by worker 

### 18 larvae at the same age. Therefore, the food consumption rate of 

### 19 queen larvae at day 5 is higher than that of the worker larvae 

### 20 at day 5. 

### 21 When we convert the honey consumption rates of 

### 22 worker and drone larvae to a nectar equivalent basis, the 

### 23 five-day worker larvae value is still less than the five-day 

### 24 queen larval value for consumption of royal jelly. This is a 

### 25 concern because we don't want to have exposure values that pass 

### 26 screens, but the colony may be impacted due to elevated

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1 pesticide doses received by the queen. This is a case where a 

### 2 Type II error could occur. 

### 3 Although the queen larvae consume more food 

### 4 compared to worker larvae at day 5, the two types of larvae are 

### 5 consuming different types of food. As I noted earlier, we have 

### 6 data available from, two different studies, involving pesticides 

### 7 that indicated that pesticide concentrations and royal jelly are 

### 8 at least 100 times lower compared to the food fed to nurse bees. 

### 9 On the next slide I will illustrate the impact of diet and 

### 10 relative concentrations on the different foods. 

### 11 This figure depicts the relative doses 

### 12 received by larvae through consumption of royal jelly or brood 

### 13 food at a concentration of one microgram active ingredient per 

### 14 kilogram and pollen and nectar at a concentration that's 100 

### 15 times greater. 

### 16 Although the consumption rate of queen larvae 

### 17 at day 5 is higher than worker larvae by a factor of 2.5, the 

### 18 overall dose received by the queen larvae is less than the 

### 19 worker larvae because the pesticide concentration in the queen 

### 20 larvae's food is so much lower. 

### 21 In summary, the proposed food consumption rate 

### 22 for the Tier I Exposure Assessment for larval honey bees appears 

### 23 to be conservative compared to other castes of honey bees. We 

### 24 also compared the proposed food consumption rate for larval 

### 25 honey bees -- which is depicted in the top row of this table -- 

### 26 to available food consumption rates for larvae of other species

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1 of bees, including the bumblebee, the European mason bee and the 

### 2 alfalfa leaf-cutting bee. 

### 3 These data suggest that the proposed food 

### 4 consumption rate for larval worker honey bees is representative 

### 5 or even protective compared to food consumption rates of these 

### 6 three species of bees. 

### 7 To summarize, the food consumption rate 

### 8 analysis, the proposed method assumes that at the Tier I level, 

### 9 food consumption can be based on nectar and pollen. This 

### 10 assumes that honey bees are either eating these foods directly 

### 11 or that they're eating stored forms of these foods. 

### 12 It is assumed that pesticide concentrations in 

### 13 pollen and nectar are equal. This also assumes that pesticide 

### 14 concentrations in brood food and royal jelly are less than those 

### 15 in nectar and pollen. The proposed food consumption rate for 

### 16 adult honey bees is 292 milligrams per day, based on the nectar 

### 17 foraging bee. The proposed larval food consumption rate of 120 

### 18 milligrams per day is based on five-day old larvae. 

### 19 Compared to other castes of honey bees and 

### 20 three other species of bees, these values appear to be 

### 21 conservative. With Charge Question 5, EPA, PMRA and Cal DPR are 

### 22 seeking feedback from the SAP on the proposed food consumption 

### 23 rates for the Tier I Assessment, including consideration of the 

### 24 calculations behind these values, their assumptions, their 

### 25 strengths and limitations, and their conservativeness relative 

### 26 to other castes of honey bees.

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1 In Charge Question 3, we are interested in 

### 2 feedback from the SAP on the extent to which the honey bee can 

### 3 be a surrogate for assessing risk to other species of bees. 

### 4 We're interested in feedback on the extent to which the 

### 5 available food consumption rates indicate that dietary doses 

### 6 predicted for honey bees at the Tier I assessment level will be 

### 7 protective for other species of bees. 

### 8 Now I'll move on to discuss the proposed Tier 

### 9 I Exposure Assessment Methods for estimating pesticide doses 

### 10 received by honey bees through contact or dietary exposure. The 

### 11 proposed contact dose for foliar spray applications is based on 

### 12 the maximum dose on forager honey bees from a study published by 

### 13 Koch and Weisser in 1997. 

### 14 Dietary exposure is assessed for foliar spray, 

### 15 seed treatments and soil applications. All three application 

### 16 types have different methods for estimating pesticide 

### 17 concentrations in pollen and nectar which are converted to doses 

### 18 using the proposed food consumption rates I just discussed for 

### 19 adult and larval honey bees. 

### 20 In all three methods, estimated pesticide 

### 21 concentrations in or on plant foliage of treated crops is used 

### 22 as a surrogate for pesticide concentrations in nectar and 

### 23 pollen. For foliar spray applications, the proposed approach 

### 24 involves the use of the tall grass concentration from the T-REX 

### 25 model, as a surrogate for pesticide concentrations in nectar and 

### 26 pollen.

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1 For seed treatments, the proposed 

### 2 concentration is one-milligram a.i. per kilogram in nectar and 

### 3 pollen based on EPPO's default screening concentration. EPPO 

### 4 stands for the European and Mediterranean Organization for Plant 

### 5 Protection. For soil treatments, the proposed method is based 

### 6 on a modification to the plant soil uptake model developed by 

### 7 Briggs, et al., and published in 1982 and 1983. 

### 8 This model is designed to estimate pesticide 

### 9 concentrations in plant shoots. When explaining each exposure 

### 10 method, I'll follow a similar format. I'll start out with a 

### 11 description of the proposed approached, followed by an 

### 12 evaluation of the method using relevant empirical data. I'll 

### 13 then discuss the assumptions and uncertainties associated with 

### 14 the method and then its strengths. 

### 15 In determining which methods to consider for 

### 16 the Tier I Exposure Assessment, we considered approaches that 

### 17 have been peer reviewed with an emphasis on models that have 

### 18 been vetted in a regulatory context. As I'll discuss, various 

### 19 methods were evaluated by comparing estimated exposures to 

### 20 available empirical data for honey bees, nectar and pollen to 

### 21 determine whether the estimates could be considered reasonably 

### 22 conservative. 

### 23 To compare the estimated exposures to the 

### 24 empirical measurements, all exposure values were normalized to 

### 25 an application rate of one point a.i. per acre. Empirical data 

### 26 were available from the scientific literature as well as

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1 unpublished registrant-submitted studies. In some cases, the 

### 2 identity of pesticides are disguised here and in the White Paper 

### 3 in order to maintain confidentiality of pesticides that are not 

### 4 yet registered. 

### 5 The level of detail and amount of data 

### 6 provided in each study varied from study to study. The amounts 

### 7 of data that could be used to evaluate the methods also varied, 

### 8 which led to differences in the evaluation of the 

### 9 conservativeness of the methods. 

### 10 For example, for foliar spray applications, 

### 11 the empirical data available to evaluate the proposed methods 

### 12 for estimating exposures through consumption of pollen and 

### 13 nectar were much larger than the data for estimating contact 

### 14 exposures. 

### 15 For foliar spray applications, the proposed 

### 16 method for estimating exposures to adult forager bees through 

### 17 contact is to use the highest dose from a field study published 

### 18 by Koch and Weisser in 1997. In this approach, the contact dose 

### 19 is 2.7 micrograms a.i. per bee, per one pound a.i. per acre. In 

### 20 applying this method, the actual dose is calculated by 

### 21 multiplying the single highest application rate in pounds a.i. 

### 22 per acre by 2.7. 

### 23 The study conducted by Koch and Weisser 

### 24 involved measuring the concentration of a tracer on honey bees 

### 25 soon after foraging in the treated area. Honey bees were 

### 26 collected, following applications to phacelia fields and apple

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1 orchards that were located in Germany. 

### 2 The study involved multiple field trials that 

### 3 were conducted over the course of several years. During this 

### 4 study, tracer residues were quantified on over 6,000 honey bees. 

### 5 The proposed dose represents the highest value that was 

### 6 measured during this study. We chose the highest dose rather 

### 7 than the upper bound due to some uncertainties associated with 

### 8 this data set, in particular, because the study only includes 

### 9 two crops. In our application of this methods, we want to be 

### 10 able to represent multiple crops. 

### 11 As indicated by comparing the doses observed 

### 12 in the trials on the apple and phacelia fields, the distribution 

### 13 of doses may vary from crop to crop. This figure depicts the 

### 14 frequency distribution of residues measured on honey bees 

### 15 foraging on phacelia fields over the course of the five trials 

### 16 that were conducted by Koch and Weisser. 

### 17 As you can see from this figure, 43 percent of 

### 18 the honey bees analyzed had residues that were in order of 

### 19 magnitude below the proposed dose of 2.7 micrograms a.i. per 

### 20 bee, normalized to one pound a.i. per acre. 

### 21 This figure depicts the frequency distribution 

### 22 of residues that were measured on honey bees foraging on apple 

### 23 orchards over the course of the nine trials that were conducted 

### 24 during this study. As you can see from this figure, 71 percent 

### 25 of all of the honey bees that were analyzed had residues that 

### 26 were in order of magnitude below the proposed dose of 2.7.

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1 These results, along with those from the 

### 2 phacelia trial suggest that the proposed dose is conservative 

### 3 when considering all of the results from this study. In 

### 4 developing the proposed method for assessing contact exposures 

### 5 to honey bees following foliar application, we also considered 

### 6 the utility of the upper bound arthropod exposure value that is 

### 7 in the current version of the T-REX model. 

### 8 This dose is 12 microgram a.i. per bee, per 

### 9 one pound a.i. per acre, which is a factor of five higher than 

### 10 the proposed contact dose from the Koch and Weisser dataset. 

### 11 This dose, the T-REX dose is calculated by multiplying the upper 

### 12 bound concentration of 97 milligrams active ingredient per 

### 13 kilogram of arthropods by the average body weight of an adult 

### 14 worker bee, which is .128 grams. 

### 15 The upper bound pesticide concentration for 

### 16 arthropods used in the T-REX model represents a 95th percentile 

### 17 concentration on arthropods, based on empirical data from the 

### 18 scientific literature and registrant-submitted studies. These 

### 19 studies measured pesticide residues on various arthropods, 

### 20 including moth and beetle larvae, crickets, grasshopper, beetles 

### 21 and unidentified arthropods that were captured in treated areas. 

### 22 This dataset does not include residue data for honey bees. 

### 23 The use of the arthropod concentration relies 

### 24 upon measured residues of pesticides in and on arthropods that 

### 25 were located on treated fields at the time that the field was 

### 26 sprayed. It is assumed that the residues are predominately

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1 based on direct spray; however, they may also incorporate some 

### 2 contact of arthropods with pesticide residues on treated 

### 3 foliage, as well as consumption. 

### 4 We chose not to propose the use of the T-REX 

### 5 arthropod dose because it does not include residues from honey 

### 6 bees. Since the focus of the Tier I risk assessment method is 

### 7 on honey bees, the proposed dose was based on the Koch and 

### 8 Weisser data. 

### 9 In cases where it is necessary to estimate 

### 10 exposures to other species of bees, the T-REX approach may be 

### 11 more representative of residues resulting from contact exposure. 

### 12 13 A review of the scientific literature failed 

### 14 to identify maximum doses following contact exposures of honey 

### 15 bees to pesticides resulting from foliar spray applications. 

### 16 Two studies were identified that contained mean residues of 

### 17 pesticides on bees. 

### 18 The figure depicted on this slide compares the 

### 19 mean doses reported in those studies to the mean doses from the 

### 20 Koch and Weisser study on phacelia and apple orchards, as well 

### 21 as the T-REX arthropod dose. The mean doses all appear to be 

### 22 similar, generally being within an order of magnitude of the 

### 23 Koch and Weisser data. 

### 24 The method evaluation for the proposed contact 

### 25 exposure dose is uncertain because there are a limited number of 

### 26 studies available to evaluate the approach. Of the two studies

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1 that are available, only mean measured concentrations on honey 

### 2 bees are reported. 

### 3 In addition, because there are only two 

### 4 studies available for evaluation purposes, it's unclear how 

### 5 representative these data are related to other locations, crops, 

### 6 and field conditions. 

### 7 It's unknown whether the data from these two 

### 8 studies are representative of high-end exposures that other 

### 9 honey bees may receive under different conditions. Despite 

### 10 these limitations, the Koch and Weisser maximum dose is 

### 11 empirically based and appears to be consistent with the data 

### 12 from the other two studies. 

### 13 Also, the proposed dose appears to be 

### 14 conservative when we consider the overall distributions of 

### 15 residues that were detected by the authors over the course of 

### 16 the study, namely that the majority of the contact doses 

### 17 measured on the foraging honey bees where in order of magnitude 

### 18 lower than the highest dose. 

### 19 The Koch and Weisser study appears to have a 

### 20 robust study design. The use of the tracer facilitated 

### 21 measurements of the full distribution of concentrations of the 

### 22 chemical on honey bees. This would not necessarily be the case 

### 23 for a fast-acting insecticide, where honey bees with lethal 

### 24 doses would not return to the hive and would not be measured. 

### 25 In addition, this dose is based on a large 

### 26 end. Each trial quantified doses on hundreds of honey bees for

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1 a total of more than 6,000 measurements on individual bees over 

### 2 the course of several years. In addition, the proposed contact 

### 3 dose is consistent with other approaches, including the T-REX 

### 4 arthropod dose as well as the Atkins Method. 

### 5 For dietary exposure, the proposal includes 

### 6 the use of an upper bound residue value for foliage that is used 

### 7 in the T-REX model. T-REX is already used by the EPA for 

### 8 ecological risk assessments of pesticides for establishing 

### 9 screening level exposures to terrestrial vertebrates. 

### 10 T-REX includes pesticide concentrations on 

### 11 different plants or plant parts, including short and tall grass, 

### 12 broadleaf plants, fruit, pods and seeds. The upper bound and 

### 13 mean concentrations used in T-REX are provided in this table. 

### 14 These concentrations are based on a review of pesticide residues 

### 15 on plants that was conducted by Kenega and then later updated by 

### 16 Fletcher, et al. 

### 17 Ideally, the proposed approach would be to 

### 18 base the upper bound concentrations on measured values from 

### 19 pollen and nectar; however, there was insufficient amount of 

### 20 empirical data to confidently estimate the upper bound. 

### 21 Therefore, it's necessary to use a surrogate. 

### 22 This figure depicts the method evaluation for 

### 23 the T-REX pesticide concentrations for short grass, broadleaf 

### 24 plants, tall grass and fruit pods and seeds. The estimated 

### 25 concentrations are depicted in this figure with red dashed 

### 26 lines. And you can see to the right of the dash which

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1 particular residue from T-REX is represented by the specific 

### 2 line. 

### 3 The empirical data that are relevant to this 

### 4 method are depicted by blue columns. I'll use this format in 

### 5 other figures throughout the rest of this presentation. 

### 6 The empirical data that are depicted here for 

### 7 nectar includes mean concentrations. Some maximum values are 

### 8 also available. Most often, only mean concentrations were 

### 9 reported in the literature. 

### 10 As you can see by this figure, the mean 

### 11 concentrations for short grass, broadleaf plants and tall grass 

### 12 are all greater than the mean concentrations available for 

### 13 nectar. The highest empirical measurement from these studies 

### 14 was 13.6 milligram per kilogram nectar, normalized to one pound 

### 15 a.i. per acre. This value is on the same order of magnitude as 

### 16 the mean concentrations for tall grass, short grass, and 

### 17 broadleaf plants. 

### 18 Four of the ten mean concentration of the 

### 19 empirical dataset exceeded the mean T-REX residue value for 

### 20 fruit, seeds, and pods. All of the empirical concentrations 

### 21 from the maximum dataset were below the T-REX upper bound 

### 22 concentrations for all four categories. 

### 23 This figure depicts the mean empirical 

### 24 concentration data for pollen, compared to the mean T-REX 

### 25 concentrations. The mean concentration for short grass is 

### 26 greater than the mean concentrations available for all of the

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1 dataset for pollen. The mean concentrations for broadleaf 

### 2 plants and tall grass exceed all but one of the nine mean 

### 3 empirical concentrations for pollen. 

### 4 Five mean pollen concentrations exceed the 

### 5 mean T-REX concentration for fruit, seeds, and pods. All of the 

### 6 maximum empirical concentrations for pollen are below the upper 

### 7 bound concentrations for short grass, broadleaf plants and tall 

### 8 grass. Seven of the 14 maximum concentrations exceed the upper 

### 9 bound concentration for fruit pods and seeds. 

### 10 So when considering all of the empirical data 

### 11 that are available for nectar and pollen, the T-REX 

### 12 concentrations for short grass, broadleaf plants and tall grass 

### 13 are consistently above the empirical concentrations. Only one 

### 14 value out of more than 40 exceeds the tall grass and broadleaf 

### 15 plant concentrations. 

### 16 The tall grass upper bound concentration is 

### 17 proposed as a surrogate because of the three possible surrogates 

### 18 that appear to be conservative for nectar and pollen. Tall 

### 19 grass is the closest to the empirical data. 

### 20 For the Tier I Exposure Assessment, the upper 

### 21 bound concentration of 110 milligram a.i. per kilogram, 

### 22 normalized to one pound a.i. per acre, would then be converted 

### 23 to a dietary dose received by adult and worker bees using pollen 

### 24 and nectar consumption rates for these two life stages. 

### 25 Therefore, the dietary-based dose for larvae is 13 microgram 

### 26 a.i. per bee, per one pound a.i. per acre, and the value for

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1 adults is 32. 

### 2 Estimated doses can be scaled to an 

### 3 application rate by multiplying by the maximum single 

### 4 application rate. And in practice, the concentrations generated 

### 5 by T-REX also account for multiple applications and dissipation 

### 6 of the chemical in between applications. 

### 7 In terms of assumptions and uncertainties 

### 8 associated with this method, I've already referenced the most 

### 9 significant one, which involves the assumption that the upper 

### 10 bound concentration for tall grass will be an appropriate 

### 11 surrogate for pesticide concentrations in pollen and nectar of 

### 12 flowers that were directly sprayed with the pesticide. 

### 13 Comparison of empirical data to the tall grass 

### 14 mean and upper bound concentrations suggests that this value is 

### 15 reasonably conservative. As empirical measures of pesticide 

### 16 concentrations on pollen and nectar become available, we may be 

### 17 able to refine the Tier I Exposure Assessment Method using 

### 18 pollen or nectar-specific data rather than using plant foliage 

### 19 as a surrogate. 

### 20 Also, in using the T-REX concentration, it's 

### 21 assumed that direct spray onto foliage represents a conservative 

### 22 estimate of concentrations in pollen and nectar for both 

### 23 systemic and non-systemic pesticides. 

### 24 Although systemic transport of pesticides to 

### 25 pollen and nectar are expected, concentrations in nectar and 

### 26 pollen that occur due to systemic transport are not expected to

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1 exceed the concentrations from direct spray. This is because 

### 2 systemic transport takes time and not all of the amounts of the 

### 3 pesticide that is applied to the plant will move the pollen and 

### 4 nectar due to dissipation of the pesticide and transport to 

### 5 other portions of the plant. 

### 6 The proposed Tier I Exposure Method for seed 

### 7 treatments is based on the EPPO 2010 screening concentration of 

### 8 one milligram a.i. per kilogram and pollen and nectar of plants 

### 9 to which seed treatments were made. EPPO screening 

### 10 concentration of one milligram a.i. per kilogram is based on the 

### 11 maximum value from data compiled by Alix, et al. in 2009, 

### 12 including pesticide residues measured in different plant parts 

### 13 following applications to soil or seed treatments. 

### 14 Plant parts included in this analysis were 

### 15 described as leaves, fruit, green parts -- in parenthesis -- 

### 16 whole plants and grains. This approach may be used for all 

### 17 pesticides that are applied via seed treatment, by assuming that 

### 18 the upper bound concentration for pollen and nectar is one 

### 19 milligram a.i. per kilogram with no need for adjustment, based 

### 20 on application rate or chemical properties. 

### 21 This concentration can then be multiplied by 

### 22 the food consumption rates for adult and larval worker bees to 

### 23 determine the upper bound doses potentially received by honey 

### 24 bees. For adults, this dose is .29 microgram a.i. per bee. And 

### 25 for larvae, the dose is .12. 

### 26 EPPO screening concentration was evaluated

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1 using empirical data describing pesticide concentration in 

### 2 pollen and nectar of crops whose seeds were treated with the 

### 3 pesticide. The highest concentration measured in pollen was 

### 4 .036 milligram a.i. per kilogram, and again, no adjustment was 

### 5 made for application rate here. This value is a factor of 28 

### 6 lower than the EPPO screening value. 

### 7 Considerably less empirical data are available 

### 8 to compare the EPPO one milligram a.i. per kilogram screen to 

### 9 nectar concentrations. The screening concentration is three 

### 10 orders of magnitude above the highest concentration for nectar, 

### 11 which is .003 milligram a.i. per kilogram. 

### 12 In the Tier I approach, it's assumed that all 

### 13 pesticides that are applied to seeds are systemic, and therefore 

### 14 can be transported into pollen and nectar that may be consumed 

### 15 by honey bees. If this is not the case for a specific 

### 16 pesticide, registrants can submit data to refute the assumption 

### 17 of systemicity. 

### 18 The proposed use of the EPPO screening level 

### 19 exposure concentration of one milligram a.i. per kilogram for 

### 20 pollen and nectar of seed treated crops does not account for 

### 21 differences in application rate where the mass of the pesticide 

### 22 that's applied to a seed. 

### 23 There is uncertainty in not accounting for the 

### 24 mass of the chemical applied since it's expected that the 

### 25 magnitude of the pesticide concentration of the plant will be 

### 26 influenced by the magnitude of the application. However, the

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1 evaluation of this screen indicates that it is protected for 

### 2 rates that are currently being used for seed treatments which 

### 3 are approximately one milligram of active ingredient per seed. 

### 4 Even if seed treatments would exceed one 

### 5 milligram a.i. per seed, it seems unlikely that pesticide 

### 6 concentrations and nectar and pollen would exceed the EPPO 

### 7 screening value since it is a factor of 28 greater than the 

### 8 highest residue that's measured in pollen and nectar. 

### 9 In addition, pesticide mass applied to seeds 

### 10 is not expected to be that much greater than one milligram a.i. 

### 11 per seed, since this value is already similar to the overall 

### 12 weight of the seed itself. 

### 13 Another uncertainty associated with this 

### 14 approach is that it does not account for physical chemical 

### 15 properties of a pesticide that may influence its potential 

### 16 systemic transport throughout the plant and into pollen and 

### 17 nectar. For example, a chemical with low mobility may be 

### 18 expected to have lower concentrations and nectar when compared 

### 19 to a chemical with higher mobility. 

### 20 Despite these limitations, this approach 

### 21 appears to generate reasonably conservative estimates of 

### 22 pesticide concentrations in the pollen of seed treated crops. 

### 23 The one milligram a.i. per kilogram screening value is much 

### 24 greater than the data that are available for nectar; however, 

### 25 there are only a few empirical concentrations, and so it may not 

### 26 be appropriate to conclude at this time that this method is

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1 unreasonably conservative for nectar. 

### 2 For soil treatments, the proposed method 

### 3 involves the use of the Briggs soil plant uptake model. This 

### 4 model predicts the concentration of a chemical in the shoot of a 

### 5 plant, using the Kow that chemical and its concentration in the 

### 6 water that is in contact with the plant's roots. 

### 7 This approach is based on observations from 

### 8 studies conducted by Briggs et al. that involved uptake of 18 

### 9 different pesticides into the shoots of barley plants. These 

### 10 pesticides included carbamate insecticides and phenylurea 

### 11 herbicides with log Kow that ranged from -.57 to 4.6. 

### 12 Note that one of the major components of the 

### 13 Briggs equation is the transpiration stream concentration factor 

### 14 or TSCF, which is a ratio of the concentration of the chemical 

### 15 in the xylem of the plant to the concentration of the aqueous 

### 16 medium that's in contact with the roots. 

### 17 Median TSCF values are calculated in the 

### 18 Briggs' published model, using the Kow of the chemical. 

### 19 Suggestions to modify the Briggs approach were 

### 20 made by Ryan, et al. in 1988. This involved adding an 

### 21 equilibrium partitioning component based on the chemicals Koc 

### 22 that allows for estimation of pesticide concentrations in 

### 23 plants, using concentrations in soil rather than in aqueous 

### 24 medium, as was used by Briggs. 

### 25 Note that if Koc is not available or 

### 26 appropriate for a chemical, the Kd can be used as an alternative

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1 to the Koc and Foc component of this equation. The soil 

### 2 properties that are required by this equation can be easily 

### 3 obtained from EFED's standard scenarios that are used for the 

### 4 prism model. 

### 5 The TSCF calculation provided by Briggs 

### 6 generates a median estimate of the TSCF in resulting 

### 7 concentration in plant shoots. As I noted at the beginning of 

### 8 this talk, the goal of the Tier I Exposure Assessment is to 

### 9 derive exposure estimates that represent an upper bound. To 

### 10 accomplish this, we recalculated TSCF values to represent the 

### 11 95th percentile values and they're depicted in this figure by 

### 12 the red line. 

### 13 These TSCF values are provided in Appendix 5 

### 14 of the White Paper and can be selected using the Kow of a 

### 15 chemical. Empirical data for pollen were available for four 

### 16 pesticides applied to pumpkin, squash, tomato or melon. 

### 17 Comparison of estimated concentrations to these measured data 

### 18 indicates that model predictions are generally conservative 

### 19 compared to empirical data. There are three empirical 

### 20 concentrations that are higher than the estimated values, but 

### 21 they are still within an order of magnitude. 

### 22 It should be noted here that some 

### 23 discrepancies between the estimated and empirical data may 

### 24 originate from several sources. For instance, none of the 

### 25 empirical data were from barley, which is the basis for the 

### 26 Briggs model. And the soil properties that are used in the

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1 Briggs model were not matched to the studies that were used to 

### 2 generate the empirical data. 

### 3 In addition, the empirical data represents a 

### 4 combination of parent and degradates, whereas the estimated 

### 5 concentrations are based on parent only. When comparing 

### 6 empirical concentrations for nectar to the estimated 

### 7 concentrations, the results are similar to those of the pollen 

### 8 analysis. Empirical concentrations for nectar were available 

### 9 for five pesticides that were applied to fuchsia, nasturtium, 

### 10 pumpkin, squash, cotton, and melon. 

### 11 Comparison of estimated concentrations to 

### 12 empirical data indicates that the model predictions are 

### 13 generally conservative compared to the empirical data. There 

### 14 are two empirical values that are higher than the estimated 

### 15 values, but they are still within an order of magnitude. As I 

### 16 noted previously, the discrepancies between the estimated 

### 17 empirical values may be attributed to differences in soil 

### 18 properties and systemic transport of the crops that are 

### 19 different than barley. 

### 20 In addition, in the case of imidacloprid, for 

### 21 which the estimated value is below one of the eight empirical 

### 22 concentrations, the estimated value is based on the parent 

### 23 alone, whereas the empirical values represent parent plus 

### 24 degradates. 

### 25 In developing the proposed method for 

### 26 estimating pesticide concentrations in nectar and pollen of

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1 crops that receive soil treatments of pesticides, we also 

### 2 considered using the EPPO one milligram a.i. per kilogram 

### 3 screening concentration that's proposed for use with seed 

### 4 treatments. When the one milligram a.i. per kilogram default is 

### 5 compared to maximum concentrations from available studies where 

### 6 pesticides were applied to soil, all empirical concentrations 

### 7 for pollen are below the one milligram a.i. per kilogram value. 

### 8 Note that this comparison did not involve 

### 9 adjustment of the concentrations for application rate. There is 

### 10 one concentration for nectar that exceeds the screen by a factor 

### 11 of almost five; however this concentration was from a study 

### 12 where the application rate of dimethoate was equivalent to 17 

### 13 pounds a.i. per acre, which is much higher than currently 

### 14 registered application rates for this chemical and most other 

### 15 insecticides in North America. 

### 16 With the exception of the dimethoate data, all 

### 17 the measured concentration of pesticides in pollen and nectar of 

### 18 plants receiving soil applications are two orders of magnitude 

### 19 below the one milligram a.i. per kilogram value. Based on this 

### 20 information, it appears that the one milligram a.i. per kilogram 

### 21 default concentration is a reasonably conservative estimate for 

### 22 soil applications. However, for pesticides that have high 

### 23 applications, this method may under-predict the exposure. 

### 24 In addition, this approach does not account 

### 25 for the physical chemical properties of a pesticide that may 

### 26 influence its potential systemic transport throughout the plant

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1 and into pollen and nectar. For example, a non-ionic organic 

### 2 chemical with low mobility may be expected to have lower 

### 3 concentrations in nectar when compared to a chemical with higher 

### 4 mobility. 

### 5 There are several notable limitations to using 

### 6 the modified Briggs approach. As with the use of the T-REX tall 

### 7 grass upper bound concentration for foliar spray applications, 

### 8 this approach assumes that estimated concentrations of a plant 

### 9 pesticide and plant foliage is an appropriate surrogate for 

### 10 concentrations in pollen and in nectar. 

### 11 Comparison of empirical data to the estimated 

### 12 concentrations suggests that the Briggs approach is reasonably 

### 13 conservative, relative to concentrations that have been measured 

### 14 in pollen and nectar. In the Tier I approach, it's assumed that 

### 15 all chemicals may be systemically transported. This assumption 

### 16 may be limited based on a chemical's log Kow, -for example, for 

### 17 chemicals that have log Kow values above five, they may not be 

### 18 systemically transported. 

### 19 But whether a chemical is transported, 

### 20 systemically in plants, could potentially be confirmed using 

### 21 empirical data that is submitted to EPA; for example, plant 

### 22 metabolism studies. However, it would be up to pesticide 

### 23 registrants to submit sufficient data to demonstrate that a 

### 24 pesticide is not systemic. 

### 25 Another limitation is that this methodology is 

### 26 based on empirical data from only one type of plant. Uptake may

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1 be different for different types of plants. However, the 

### 2 species that was used by Briggs, which is barley, is 

### 3 representative of a crop species that is likely to be grown on 

### 4 soils where pesticides are applied. 

### 5 Also, the dataset used to establish the Briggs 

### 6 equation is based on a limited number of chemicals that 

### 7 represent only two classes of pesticides. Despite this 

### 8 limitation, the pesticides that were used in this approach are 

### 9 systemic and cover a wide range of log Kow values, leading to an 

### 10 approach that is applicable to a wide range of pesticides with 

### 11 the potential to be systemically transported by plants. 

### 12 However, the approach may have limited utility for ionic 

### 13 chemicals whose transport may not be predicted while using Kow 

### 14 and Koc. 

### 15 The final limitation I'll discuss is that the 

### 16 proposed approach is based on passive transport of chemicals 

### 17 into the xylem. 

### 18 This approach does not directly estimate 

### 19 pesticide concentrations in plants that are the result of phloem 

### 20 transport. One possible assumption that could be made in 

### 21 applying this model for estimating pesticide concentrations in 

### 22 plants would be to assume that chemical transport via phloem, is 

### 23 equivalent to that of xylem. 

### 24 The alternative approach would be to use 

### 25 different models for neutral and ionic compounds such as those 

### 26 described by Briggs, et al. 1987; Kleier 1988; and Trapp, et al.

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1 1995. Those citations are provided in the White Paper. 

### 2 Despite the limitations discussed here, both 

### 3 the Briggs model with the modifications by Ryan, et al. and 

### 4 those proposed by EPA in the White Paper, as well as the EPPO 

### 5 one milligram a.i. per kilogram screening value, both appear to 

### 6 be useful tools for generating reasonably conservative estimates 

### 7 of pesticide concentration in plants after systemic transport 

### 8 from the soil. These approaches are consistent with the 

### 9 screening level assessment because they're both simple and 

### 10 efficient and are empirically based on data for pesticides. 

### 11 The Briggs model had the advantage that it 

### 12 relies upon some basic physical chemical properties of 

### 13 pesticides and also accounts for the mass that's applied to the 

### 14 soil. Therefore, the Briggs model is the one that's proposed 

### 15 for estimating pesticide concentrations in pollen and nectar, 

### 16 following a soil treatment. 

### 17 This table summarizes the proposed Tier I 

### 18 Exposure Methods. The values provided in the right column 

### 19 represent the resulting doses for each method, based on an 

### 20 application of one pound a.i. per acre. For foliar 

### 21 applications, the proposed Tier I Exposure Assessment involves 

### 22 quantification of pesticide doses to adult honey bees through 

### 23 contact exposure. This method is based on the maximum dose of 

### 24 2.7 microgram a.i. per bee, per one pound a.i. per acre that was 

### 25 measured on honey bees as recorded by Koch and Weisser. 

### 26 For foliar spray applications, seed treatments

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1 and soil treatments that proposed Tier I Exposure Assessment, 

### 2 quantifies pesticide doses received by adults and larvae through 

### 3 the diet. These methods involve translating estimated 

### 4 concentrations in pollen and nectar into doses by multiplying 

### 5 the concentrations by the food consumption rates of adults and 

### 6 larvae. For adults, the proposed food consumption rate is 292 

### 7 milligrams of food per day. And for larvae, that value is 120. 

### 8 9 For foliar spray applications, the proposed 

### 10 method involves using the T-REX tall grass upper bound 

### 11 concentration of the surrogate for pesticide concentrations in 

### 12 pollen and nectar. The resulting doses of 32 micrograms a.i. 

### 13 per bee and 13 micrograms a.i. per bee for adults and larvae can 

### 14 be adjusted to the use of the pesticide by accounting for the 

### 15 application rate of the chemical. 

### 16 These doses are the highest among the proposed 

### 17 methods, indicating that if the application rate is held 

### 18 constant, then foliar spray applications would represent the 

### 19 highest potential exposures to honey bees at the Tier I Exposure 

### 20 Assessment level. 

### 21 For seed and soil treatments of pesticides, 

### 22 it's assumed that the pesticides are systemically transported to 

### 23 pollen and nectar. For seed treatments, the proposed method is 

### 24 to use the EPPO screening value of one milligram a.i. per 

### 25 kilogram to represent pesticide concentrations in nectar and 

### 26 pollen of treated crops. No adjustment s made for application

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1 rate or mass of chemical applied to the seed. Therefore, for 

### 2 all uses and chemicals, the doses would be .29 and .12 microgram 

### 3 a.i. per bee for adults and larvae respectively. 

### 4 For soil treatments, the proposed method for 

### 5 estimating pesticide concentrations in nectar and pollen 

### 6 involves the use of the modified Briggs soil plant uptake model. 

### 7 This model relies on a pesticide's application rate, Kow and 

### 8 Koc to estimate the concentration in plant shoots which is a 

### 9 surrogate for pollen and nectar. 

### 10 As an illustration of the relative doses 

### 11 generated by this approach, the values depicted in this table 

### 12 are for dimethoate, which is a mobile chemical with a low Kow. 

### 13 In evaluating the proposed methods using empirical data, the 

### 14 proposed methods appear to be reasonably conservative. 

### 15 As additional information becomes available, 

### 16 we may reevaluate the proposed methods, for example, as targeted 

### 17 monitoring data become available where pesticides are measured 

### 18 in pollen and in nectar as part of Tier II studies, it may be 

### 19 possible in the future to replace the use of pesticide 

### 20 concentrations on plant foliage as surrogates with empirical 

### 21 measures of pesticides and pollen and in nectar. 

### 22 The EPA is interested in SAP feedback on the 

### 23 proposed Tier I Exposure Assessment Method that I discussed in 

### 24 this presentation and is presented in detail in Section 3 of the 

### 25 White Paper. For contact exposures resulting from foliar 

### 26 applications, which is the focus of Question 4, EPA, PMRA and

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1 Cal DPR are interested in SAP comments on the strengths and 

### 2 limitations of the proposed approach that is based on the 

### 3 maximum of the Koch and Weisser data. 

### 4 We're also interested in SAP feedback in the 

### 5 potential utility of the T-REX arthropod dose in assessing 

### 6 contact exposure to non-Apis bees as well as the honey bee. The 

### 7 focus of Charge Question 6 is on the proposed methods, meaning 

### 8 pesticide concentration in pollen and nectar, which are relevant 

### 9 to assessing dietary exposures. 

### 10 We're requesting SAP feedback to the extent to 

### 11 which the T-REX tall grass upper bound concentration the EPPO 

### 12 one milligram a.i. per kilogram screen and the modified Briggs 

### 13 model can represent pesticide concentrations in pollen and 

### 14 nectar for foliar spray, seed treatment and soil applications 

### 15 respectively. 

### 16 We're also interested in any other methods or 

### 17 data that may represent viable alternatives to these proposed 

### 18 methods or may be used to improve the proposed methods, perhaps, 

### 19 in the form of reducing assumptions or uncertainties. 

### 20 If I have time, I can take questions. 

### 21 **DR. DANIEL SCHLENK:**Thanks. Dr. 

### 22 Fefferman. 

### 23 **DR. NINA FEFFERMAN:**Okay. Thank you. That 

### 24 was really interesting. I'm wondering, a lot of the estimates 

### 25 that I saw for how you were using characterizations of larval 

### 26 and adult uptake for consumption rates were for normal colony

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1 processes. And I'm wondering if so, how you guys are dealing 

### 2 with non-normal but lifecycle dependent colony processes. 

### 3 So gorging, having to leave the hive very 

### 4 quickly or swarming times when both the energy needs of an 

### 5 individual may be different and also the consumption rate may 

### 6 skyrocket for non-energetic process needs. 

### 7 **MS. KRISTINA GARBER:**The proposed method is 

### 8 based on kind of the baseline functioning of the colony. It 

### 9 doesn't account for those scenarios like swarming. 

### 10 **DR. NINA FEFFERMAN:**Can we stick something in 

### 11 there about that? Because if they accidentally poison 

### 12 themselves once a year that might be really bad. 

### 13 **MS. KRISTINA GARBER:**That's part of why we're 

### 14 here. So if you have any advice on how we could potentially 

### 15 modify that and some data that you could point to or good 

### 16 sources that could provide another way to quantify the 

### 17 consumption rates, that would be appreciated. 

### 18 **DR. NINA FEFFERMAN:**Sure. Thanks. 

### 19 **DR. DANIEL SCHLENK:**Okay. Dr. Berenbaum. 

### 20 **DR. MAY BERENBAUM:**So estimating the contact 

### 21 exposure doses based on models that derive from empirical 

### 22 studies that in some cases are 15 to 30 years old, is there any 

### 23 indication or is it a consideration that detection methods have 

### 24 changed so that the ability to detect residues is also a 

### 25 variable in this process? 

### 26 **MS. KRISTINA GARBER:**Just for clarification,

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1 are you talking about the Briggs model as being -- 

### 2 **DR. MAY BERNEBAUM:**That's one that was 

### 3 referenced to Atkins '82 [sic] in there. There are a couple of 

### 4 studies, Koch and Weisser from '97, I think. So these are older 

### 5 technologies. Is there some consideration of how chemical and 

### 6 analytical techniques change so that detection is improving? 

### 7 **MS. KRISTINA GARBER:**I think that at this 

### 8 point, when we were looking at these studies, I think that the 

### 9 detection rates were sufficient to confidently estimate the 

### 10 values that were being detected. 

### 11 If you have any concerns about the specific 

### 12 detection rates, some of the studies would definitely account 

### 13 for that. If that's an uncertainty in some of the methods, then 

### 14 certainly we could look at newer studies that may account for 

### 15 that uncertainty. 

### 16 **DR. DANIEL SCHLENK:**Okay. Dr. James. 

### 17 **DR. ROSALIND JAMES:**Your doses are really 

### 18 dose per day, right? 

### 19 **MS. KRISTINA GARBER:**Yes. 

### 20 **DR. ROSALIND JAMES:**I work with solitary bees 

### 21 and they make a pollen provision and it's really easy to 

### 22 calculate the total dose that larvae will have over their entire 

### 23 development period. Honey bees also only feed for five days. 

### 24 One thing that I was thinking about when you had the drone 

### 25 consumption rates and they're eating less than the workers as 

### 26 larvae and yet they're bigger, so you know they must be eating

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1 more because they have to get bigger somehow. If the 

### 2 measurements are accurate, it must be because they eat longer, 

### 3 they're larvae for longer, and similarly with queens. 

### 4 So they could potentially be exposed to higher 

### 5 amounts of pesticide as larvae. But if you're only looking at 

### 6 dose per day you would miss that, right? 

### 7 **MS. KRISTINA GARBER:**In developing this 

### 8 model, we did go back and forth between the idea of the duration 

### 9 of the exposure we were trying to represent. I think that what 

### 10 you're saying is a valid point, you know, that the duration of 

### 11 the EEC or the exposure that we're looking at, it is relevant. 

### 12 And part of why we settled on day was that it could be matched 

### 13 to the acute toxicity test when we're only exposing the bees for 

### 14 a shorter period of time. 

### 15 **DR. DANIEL SCHLENK:**Dr. Tarpy. 

### 16 **DR. DAVID TARPY:**I have a couple of 

### 17 questions, actually. One is largely out of my own ignorance, 

### 18 not being a toxicologist. But knowing that that there is 

### 19 variation, sometimes substantial variation, among workers, not 

### 20 to mention, huge variation with different castes, and even 

### 21 larger variation among different pollinators of which honey bees 

### 22 are supposed to be a proxy in this means; in the in vitro 

### 23 studies, why is the denominator a per bee, per day basis rather 

### 24 than say, milligram per individual? 

### 25 **MS. KRISTINA GARBER:**By "denominator" you 

### 26 mean the toxicity endpoint?

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1 **DR. DAVID TARPY:**Yeah. All of the 

### 2 recommendations here on the milligrams of active ingredient per 

### 3 bee. Why is it on a per individual basis rather than something 

### 4 more standardized that can translate across different life forms 

### 5 and different types of pollinators? 

### 6 **MS. KRISTINA GARBER:**I think that part of 

### 7 that is the historical basis of how honey bee toxicity tests are 

### 8 conducted where they tend to be on a dose per individual bee. 

### 9 That's how the residues are quantified. 

### 10 **DR. DAVID TARPY:**Okay. So it's just based on 

### 11 practice and historical purposes, but there is variation there 

### 12 but it's not captured in these types of bioassays. 

### 13 **MS. KRISTINA GARBER:**Just to clarify, by 

### 14 "variation," do you mean individual variability? 

### 15 **DR. DAVID TARPY:**Yeah. Bees weigh different 

### 16 from each other. 

### 17 **MS. KRISTINA GARBER:**Right. 

### 18 **DR. DAVID TARPY:**And I would presume, and 

### 19 again, correct me if I'm wrong, but that toxicity would be a 

### 20 function of weight, of body weight to some capacity, assuming 

### 21 that's even linear. It might even be nonlinear effects, right. 

### 22 23 I just wanted to ask that question, not 

### 24 knowing what the answer was. So you seem to have answered that. 

### 25 26 **MS. KRISTINA GARBER:**Okay. Presumably, you

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1 would catch some of the variability in the confidence interval 

### 2 that would surround your LD50. That would be generated from 

### 3 that. So you'd probably get -- if there is a relationship 

### 4 between weight and toxicity, then that would be translated in 

### 5 the 95th percentile confidence around your LD50. 

### 6 **DR. DAVID TARPY:**Per individual. 

### 7 **MS. KRISTINA GARBER:**Correct. 

### 8 **DR. DAVID TARPY:**Okay. The second question 

### 9 is combining the total food intake for the different adult 

### 10 caste, behavioral caste, especially with the pollen versus 

### 11 nectar, is there consideration in doing these bioassays for 

### 12 lipophilic versus hydrophilic compounds in those different 

### 13 phases that in some cases, combining those two things in total 

### 14 food consumption per day might not be captured for particular 

### 15 compounds that are being screened. 

### 16 **MS. KRISTINA GARBER:**So I have to confess, 

### 17 you're asking a lot of toxicity questions that are related to a 

### 18 talk that Joe DeCant will be giving. 

### 19 **DR. DAVID TARPY:**Okay. 

### 20 **MS. KRISTINA GARBER:**If I may, would you mind 

### 21 deferring your question to that time? Maybe that will be 

### 22 covered. 

### 23 **DR. DAVID TARPY:**Mm-hmm. No problem. 

### 24 **MS. KRISTINA GARBER:**Thank you. 

### 25 **DR. DANIEL SCHLENK:**Dr. Potter. 

### 26 **DR. THOMAS POTTER:**I had a question about any

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1 consideration that may have been made regarding physical 

### 2 chemical properties of the chemicals and the potential 

### 3 concentrations in nectar, given its importance, in terms of an 

### 4 exposure pathway. Something, for example, is the aqueous 

### 5 solubility of the chemical or its solubility in the 30 percent 

### 6 sugar solution or something to that degree. 

### 7 At some point, solubility will define an upper 

### 8 bound. Perhaps that's something that you guys thought about. 

### 9 Essentially, I'm asking, is there anything that went into your 

### 10 thought processes in that regard? 

### 11 **MS. KRISTINA GARBER:**We didn't directly 

### 12 consider using the solubility. I would think that given that 

### 13 the nectar would have more than just water, it might complicate 

### 14 the way that we address solubility, but that's something that we 

### 15 could consider in the future. But we didn't consider that as an 

### 16 alternative to the Briggs model. 

### 17 **DR. THOMAS POTTER:**I wasn't referring to the 

### 18 Briggs model, per se, but in terms of contact exposure from 

### 19 foliar sprays that are intercepted by pollen and nectar. You 

### 20 had several studies that you pulled from the literature where 

### 21 there were some measured data. 

### 22 What was interesting about those studies, and 

### 23 I'll comment on them in the question period, but they were 

### 24 looking at -- a couple of the chemicals that were extremely 

### 25 insoluble in water. So you wouldn't expect to be finding them 

### 26 in the nectar.

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1 I had some concerns about one of the 

### 2 particular studies. Also about their analytical techniques as 

### 3 well. But I think it's an important consideration and without 

### 4 too much effort, I think you could put some thought into that 

### 5 and bring that into the process. I think it would strengthen it 

### 6 and certainly add to its credibility. 

### 7 **DR. THOMAS STEEGER:**I would just like to add, 

### 8 part of the risk assessment process that we'll be talking about 

### 9 later on this afternoon is the risk characterization component. 

### 10 Part of that is the risk discussion and just risk description 

### 11 section of risk characterization and in there the risk assessor 

### 12 would attempt to capture solubility issues and how it relates to 

### 13 screening level assumptions so that those types of uncertainties 

### 14 are indeed characterized in our risk assessments. 

### 15 **DR. THOMAS POTTER:**What you're saying is 

### 16 you're catching that downstream then? 

### 17 **DR. THOMAS STEEGER:**To the best of our 

### 18 ability. 

### 19 **DR. THOMAS POTTER:**Okay. 

### 20 **DR. THOMAS STEEGER:**Okay. Thanks. 

### 21 **DR. DANIEL SCHLENK:**Dr. Sandy. 

### 22 **DR. MARTHA SANDY:**I have a question about the 

### 23 estimate of contact exposures for the foliar spray applications. 

### 24 So you were basing that on a study with two crops, apples -- I 

### 25 have to admit I don't know what that other crop was, phacelia or 

### 26 whatever. So I'm wondering, how representative are those two

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1 crops to other crops, like corn and maybe there are others that 

### 2 bees might come into more contact in the process of gathering 

### 3 nectar and pollen. 

### 4 **DR. THOMAS STEEGER:**The use of phacelia, it's 

### 5 a European plant and it has come into considerable use, as of 

### 6 late, in the semi-field studies, intent on doing screening level 

### 7 studies is to use a crop that is particularly pollinator 

### 8 attractive and is a good source of both pollen and nectar. 

### 9 So phacelia fits into that category well. It 

### 10 has a robust bloom. So presumably, based on it being pollinator 

### 11 attractive, it is, as I say, presumed to be relatively 

### 12 protective and a source of information regarding residues that 

### 13 could accumulate in pollen and nectar, either through a foliar 

### 14 application or because of systemic transfer to pollen and 

### 15 nectar. 

### 16 **DR. DANIEL SCHLENK:**Dr. McManaman. 

### 17 **DR. JAMES MCMANANMAN:**I have a question of 

### 18 clarification, basically because I'm not a bee physiologist, but 

### 19 your pesticide dose used for the larvae, you claim that the 

### 20 pollen and nectar have pesticide concentrations, about 100 times 

### 21 higher in pollen and nectar compared to royal jelly. Is that 

### 22 correct? 

### 23 **DR. THOMAS STEEGER:**That is correct. 

### 24 **DR. JAMES MCMANAMAN:**I think that your Tier 

### 25 I, for larvae testing, you're planning on using Day 5 larvae. 

### 26 What I've read from the literature over the course of the past

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1 couple of weeks is that Day 5 larvae still consume royal jelly. 

### 2 I mean, it certainly would be more conservative, but would that 

### 3 be inaccurate using those levels to test, would that actually 

### 4 give result that really wouldn't be applicable to the real 

### 5 world? 

### 6 **MS. KRISTINA GARBER:**I think that is an 

### 7 uncertainty associated with the proposed approach that we are 

### 8 assuming that they would be eating honey and pollen only. We 

### 9 didn't have data available to quantify how much royal jelly they 

### 10 may be consuming at that time. 

### 11 So as a conservative approach, we chose to 

### 12 represent the entire diet as pollen and nectar. So that may 

### 13 over-predict, but honestly, we don't have any data to tell us 

### 14 how much. 

### 15 **DR. JAMES MCMANAMAN:**Of the bee experts in 

### 16 the room, do they consume royal jelly or do they consume pollen 

### 17 at Day 5 larvae? 

### 18 **UNIDENTIFIED SPEAKER:**Brood food. 

### 19 **DR. JAMES MCMANAMAN:**Pollen? Brood food. 

### 20 Okay. All right. So it's accurate then. Good. 

### 21 **DR. DANIEL SCHLENK:**Dr. Hunt. 

### 22 **DR. GREG HUNT:**Maybe this will be clarified 

### 23 later, but with -- it's suggested to use .29 micrograms for seed 

### 24 treatment products. For clothianidin, that represents about 70 

### 25 times the LD50 for the smallest estimate of the LD50. 

### 26 Given that this is already being used in corn,

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1 I assume that means that it needs to go to Level II trials and 

### 2 Level III trials. I'm wondering, what are the ramifications? 

### 3 How long will that take? I'll just throw that out there for 

### 4 whoever wants to respond. 

### 5 **DR. THOMAS STEEGER:**Without getting into how 

### 6 this actually plays out in terms of data requirements, as you've 

### 7 pointed out, for a chemical that has failed a Tier I screen, 

### 8 first, the decision has to be made by the risk manager whether 

### 9 to ask for additional information. And the process is 

### 10 management-based process as opposed to a risk assessment-based 

### 11 process. 

### 12 So the higher tier studies, it depends on the 

### 13 nature of the study and it depends on the results, how far you 

### 14 move up in the refinement process. I don't want to digress too 

### 15 much here, but depending on the risk assessment question or the 

### 16 risk management question that's being asked at the higher tier 

### 17 will dictate the length of the study, particularly if you want 

### 18 to know what effect overwintering; what effect the chemical may 

### 19 have overwintering, these can be long studies. 

### 20 The bottom line is that we can't rush the 

### 21 science to derive an answer that involves regulating pesticides 

### 22 at a national level. The foundation of our risk assessments has 

### 23 to be quality science. That being said, it can take a while for 

### 24 quality science to be generated. I know that's frustrating, but 

### 25 that's the reality that we have to deal with in terms of being a 

### 26 regulatory agency that's founded on quality science.

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1 **DR. DANIEL SCHLENK:**Okay. Any other 

### 2 questions or clarification on the presentation? 

### 3 (No response.) 

### 4 Okay. Let's go ahead with the next 

### 5 presentation which will be by Reuben Baris. 

### 6 **MR. REUBEN BARIS:**Good afternoon. My name is 

### 7 Reuben Baris. I'm an environmental scientist in the 

### 8 Environmental Fate and Effects Division of the Office of 

### 9 Pesticide Programs. Thank you very much for the opportunity to 

### 10 address the SAP. 

### 11 I will be talking about methods for 

### 12 characterizing Tier I exposures to bees, as well as study design 

### 13 and objectives for Tier II exposure studies. These topics are 

### 14 presented in detail in Section III, and in part, Section IV of 

### 15 the White Paper. 

### 16 The studies that I will present in the first 

### 17 part of this presentation are currently included in the Part 158 

### 18 data requirements. With minor adjustments and modifications, 

### 19 these studies have the potential to be used to refine exposure 

### 20 estimates. 

### 21 As discussed previously, the risk assessment 

### 22 process is intended to be iterative. At the Tier I level, 

### 23 exposure value is used to estimate risk quotients are intended 

### 24 to be conservative. However, for chemicals identified to be of 

### 25 concern at the first tier, refined estimates of exposure may be 

### 26 needed to provide more realistic understanding of pesticide

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1 exposures to bees under actual use conditions. 

### 2 Moving from Tier I to Tier II is a function of 

### 3 risk management needs and decisions. Tier II studies can be 

### 4 solely measures of exposure as pollen and nectar or can be 

### 5 coupled with measures of exposure and effects. Higher tiered 

### 6 studies on the effects of pesticides on honey bees will be 

### 7 presented by Mr. DeCant, following this presentation. 

### 8 I would like to return your focus to the 

### 9 proposed decision tree, specifically, seen here in Box 6, 

### 10 Refining the Tier I Exposure Assessment. I will touch on a few 

### 11 studies that are currently available, where with modern 

### 12 modifications to the protocols could be used to refine Tier I 

### 13 exposure estimates and recalculate risk quotients. The effects 

### 14 data will not change, but with these studies, we are able to 

### 15 refine estimates of dietary exposure. 

### 16 For pesticides, they are applied as a foliar 

### 17 spray and exceed levels of concern for dietary exposure to bees. 

### 18 It may be possible to refine dietary-based estimates of 

### 19 exposure. 

### 20 This refinement could be accomplished using 

### 21 pesticide-specific measurements of treated foliage taken within 

### 22 24 hours of application. Registrant-submitted magnitude of 

### 23 residue studies used by the Health Effects Division may be used 

### 24 for this type of refinement. 

### 25 The empirical studies presented on the next 

### 26 slide may contain information on the transport of a compound in

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1 the plant. That is, systemic or non-systemic transport; 

### 2 therefore, will influence the need for higher tiered studies. 

### 3 For example, if the chemical in soil is applied and not detected 

### 4 in plant tissue, fruit, or flower, it is not likely systemic. 

### 5 Therefore, exposure is unlikely and higher tiered studies would 

### 6 not be required. 

### 7 The table displayed here outlines empirical 

### 8 data where identified to be potentially useful for refining Tier 

### 9 I exposure estimates. These data are routinely submitted as 

### 10 part of registration or could be submitted if conditionally 

### 11 required. The Xs represent which study could be used to refine 

### 12 Tier I estimates of exposure, based on the application type. 

### 13 None of the studies listed are specifically 

### 14 designed to measure high-end pesticide concentrations in pollen 

### 15 and nectar at times when bees may be exposed. With modification 

### 16 to the study design, these studies may be used to generate 

### 17 chemical-specific measures of pesticides in pollen and nectar 

### 18 that could be used for chemicals that do not pass the initial 

### 19 Tier I screen. Some of these studies may be used as refinements 

### 20 for seed treatments as well. 

### 21 With some limitations recognized, the studies 

### 22 listed here are currently Part 158 data requirements for 

### 23 registering pesticides and may be used, with minor 

### 24 modifications, to study protocols to obtain Day 0 data, or at 

### 25 the very least, could be used as confirmatory data for 

### 26 determining the systemic nature of the compound in plants.

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1 Again, I would like to bring your attention 

### 2 back to the proposed decision tree. For the second portion of 

### 3 my talk, I will be referring to Box 9(a), Conducting Tier II 

### 4 Exposure Studies. Box 9(a) is intended for chemical-specific 

### 5 residue studies as presented earlier by Mr. Sappington. 

### 6 The purpose of Box 9(a) and the intent of the 

### 7 Tier II Exposure Assessment is to obtain pesticide-specific, 

### 8 empirically-based exposure data that represent concentrations 

### 9 encountered by bees. Like the Tier I exposure method, potential 

### 10 exposures should be assessed through quantification of doses 

### 11 through direct spray or diet. 

### 12 Specifically, studies should be designed to 

### 13 quantify pesticide residues in pollen and nectar on bees 

### 14 foraging in the treated field. The dotted line depicted in this 

### 15 figure shows the process where the Tier II exposure values will 

### 16 be used to refine Tier I exposure estimates for adults and brood 

### 17 and then recalculate risk quotients. 

### 18 Tier II exposure studies with bees are not 

### 19 typically conducted in the laboratory, but rather focus on 

### 20 relatively controlled field studies where the movement of bees 

### 21 to and from the colony is restricted through the use of 

### 22 enclosures containing the treated crop. These studies may be 

### 23 designed to measure residues on parent only or parent and 

### 24 potential degradates of concern. 

### 25 The next few slides describes the basic 

### 26 objectives and design elements of the two types of field studies

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1 that can be used to quantify pesticide exposure and matrices 

### 2 that are relevant to bees, such as targeted field studies and 

### 3 semi-field tunnel studies. 

### 4 The higher tiered studies are designed to be 

### 5 conducted under vulnerable targeted conditions to provide 

### 6 quantifiable, refined estimates of pesticide concentrations in 

### 7 bee food sources. 

### 8 The strength of basing the Tier II exposure 

### 9 approach on empirical data from field studies is that some of 

### 10 the uncertainties associated with the Tier I exposure method are 

### 11 reduced or eliminated. For instance, the assumption 

### 12 incorporated into the Tier I Exposure Assessment for foliar 

### 13 applications that pesticide concentrations are in grass are 

### 14 equivalent to nectar and pollen, is not necessary if pesticide 

### 15 concentrations are quantified directly in pollen and nectar. 

### 16 Also, the Tier I use of upper bound exposure 

### 17 values is based on a large set of chemicals that is not 

### 18 necessary for pesticide-specific residues, can be estimated 

### 19 under field conditions that are representative of the 

### 20 pesticide's intended use pattern. 

### 21 As part of the tiered testing process for 

### 22 evaluating potential exposures and effects on bees, higher 

### 23 tiered refinements shift the focus from exposure and effects on 

### 24 individual bees to that of the intact colony. Typical designs 

### 25 of semi-field studies include a crop that is grown outdoors in 

### 26 an enclosed system with controlled or confined exposure.

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1 The test crop is grown under good agricultural 

### 2 practices, receiving the maximum-labeled application rate, with 

### 3 a minimum interval between applications. The test crop provides 

### 4 the source of pollen and nectar. The design could be structured 

### 5 to reflect a desired exposure system or foraging environment 

### 6 such as the mixture of crops and weeds, or flowering margins, 

### 7 although, such designs can introduce additional variables which 

### 8 may be difficult to control across replicates and treatments. 

### 9 As is evident, there are a range of different 

### 10 designs and the actual test structure and study design will 

### 11 depend on the specific questions asked in the risk assessment in 

### 12 coordination with risk management needs. Ultimately, the Tier 

### 13 II exposure studies must be conducted in a manageable way so 

### 14 that data is collected and used to inform the risk assessment. 

### 15 The field and tunnel residue studies can 

### 16 provide important information regarding the levels of 

### 17 contamination potentially encountered by bees in the field; 

### 18 however, there are a number of uncertainties associated with 

### 19 these types of studies. One major uncertainty is that the level 

### 20 of residues inside the hive may be different than those first 

### 21 encountered by foraging bees in the field. 

### 22 In other words, pesticide concentrations in 

### 23 freshly collected nectar and pollen could differ from residues 

### 24 and honey and bee bread. Bee bread and honey, which form the 

### 25 basis of the hive diet, undergo processing by bees. 

### 26 Consequently, measures of residues in pollen and nectar

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1 collected directly from the flower or from returning bees may be 

### 2 different from the exposure to individuals within the hive. 

### 3 Tier I exposure estimates are conservative for 

### 4 unprocessed food sources. The objective of the higher tiered 

### 5 studies for exposure are dependent upon the Tier I assessment 

### 6 and where we have the ability to refine those estimates. 

### 7 Refinements are chemical specific 

### 8 concentrations in pollen and nectar that are used to refine the 

### 9 quantitative risk quotient. Another notable limitation for the 

### 10 Tier II Exposure Assessment Method is that it will require a 

### 11 substantial amount of time and resources to complete. 

### 12 In the future, it may be possible that the 

### 13 data required from targeted monitoring studies may be used to 

### 14 establish patterns of exposure based on application methods, 

### 15 crops and location. These patterns could then be used to 

### 16 develop a refined method where the data could be extrapolated to 

### 17 other chemicals. 

### 18 In addition, some field and tunnel residue 

### 19 studies that are currently being conducted also involve the 

### 20 collection of data on residues and leaves and flowers. These 

### 21 data potentially provide a more complete dataset from which to 

### 22 evaluate the ability of flowers or leaf residues to provide a 

### 23 conservative measure of residues in pollen and nectar. 

### 24 The first type of Tier II study presented is a 

### 25 targeted field study or field residue trial, where pesticide 

### 26 residues are quantified in pollen and nectar, collected directly

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1 from the treated crop. This study could be specifically 

### 2 designed with the sole intent of quantifying pesticide residues 

### 3 and matrices relevant to honey bees, or could be incorporated 

### 4 into the study design of an existing protocol required for 

### 5 pesticide registration, such as a cropped terrestrial field 

### 6 dissipation study. 

### 7 In the design of these studies, it is 

### 8 important to consider the type of application, that is, foliar, 

### 9 soil, or seed treatment and relate the design to the desired 

### 10 outcome. 

### 11 The second study is referred to a tunnel study 

### 12 because the treated crop is kept under an enclosure, along with 

### 13 a nucleus hive of bees, which are small colonies with roughly 

### 14 5,000 bees. Study design guidance is currently provided by 

### 15 EPPO, OACD, and also more recently by EFSA. 

### 16 The selection and use of this study, compared 

### 17 to the targeted field study, must be considered, along with the 

### 18 desired outcomes of the Tier II study. For example, if the 

### 19 desired outcome is to collect pollen, nectar, and bee residue 

### 20 data, that is, pollen sacs, pollen traps, or regurgitated 

### 21 nectar, then the semi-field study would meet those objectives. 

### 22 If the goal is to obtain targeted field data, 

### 23 that is, measured residues in pollen and nectar directly from 

### 24 plants, the targeted field study would meet those objectives. 

### 25 The difference in the goals of the Tier II studies is the 

### 26 measured residues of relevant bee matrices, where the tunnel

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1 study design is set up to measure bees and hive products. 

### 2 Additional detail for the Tier II study design will be 

### 3 presented, following this presentation. 

### 4 There are several sources of variation related 

### 5 to the anticipated use of the pesticide that should be 

### 6 considered when designing the studies for Tier II Exposure 

### 7 Assessment, including application methods, props, and location. 

### 8 Application methods employed for a chemical can vary. Because 

### 9 different application methods may result in different levels and 

### 10 types of exposure application practices that are expected to 

### 11 generate high-end exposure concentrations for pesticides should 

### 12 be selected. 

### 13 In determining which crops to include in field 

### 14 studies, a focus should be placed on registered uses for crops 

### 15 that are highly attractive to bees and that have the highest 

### 16 application rates. For systemic pesticides in particular, 

### 17 different plant species may have differences in uptake, 

### 18 distribution, and metabolism of the test substance. 

### 19 In addition, different plants have different 

### 20 matrices that represent food sources for bees. For example, 

### 21 corn only produces pollen that bees will collect as a food 

### 22 source, whereas, canola produces both nectar and pollen. Some 

### 23 plants, such as cotton, also produce extra floral nectaries 

### 24 which may be present when flowers are not. 

### 25 The sampling scheme is an important 

### 26 consideration in the design of the Tier II studies. Sampling

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1 should target the maximum concentrations found in pollen and 

### 2 nectar, therefore, these studies should sample pollen and nectar 

### 3 at different time points during bloom to evaluate the presence 

### 4 and decline of the compound over time. 

### 5 Sample timing should be selected based on the 

### 6 consideration of the dissipation of the chemical though 

### 7 degradation and transport, away from the plant, as well as 

### 8 systemic transport through the plant to pollen and nectar. For 

### 9 foliar applications, studies should be designed such that the 

### 10 pesticide concentrations are quantified on the day of 

### 11 application and during subsequent days that are selected to 

### 12 represent the concentrations on bees and relevant plant samples. 

### 13 For soil and seed treatment applications, 

### 14 sampling should account for the delayed transport from roots to 

### 15 pollen and nectar. Different field locations have different 

### 16 climates and soils, and therefore, growing conditions at a 

### 17 spacial scale. 

### 18 The locations of the studies should be 

### 19 selected so that they represent anticipated use sites of the 

### 20 chemical. If possible, honey bee specific studies should be 

### 21 conducted in multiple regions that are identified to be of 

### 22 concern. 

### 23 As a result of these considerations, it may be 

### 24 necessary to conduct targeted studies, using multiple locations, 

### 25 methods, crops, and locations in order to quantify potential 

### 26 pesticide exposures to bees under conditions that are

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1 represented of the anticipated use of the chemical. 

### 2 Because field residue data may exhibit extreme 

### 3 spacial and temporal variability, it is necessary to collect a 

### 4 sufficient amount of data to allow the risk assessor to have an 

### 5 understanding of the central tendency and upper bound exposure 

### 6 that represents the potential environmental exposures. Thank 

### 7 you very much for your time. I would like to open the floor to 

### 8 any clarifying questions the panel may have. 

### 9 **DR. DANIEL SCHLENK:**Any questions 

### 10 regarding exposure. Again, we're going to be talking about 

### 11 toxicity in the next presentation. Dr. James? 

### 12 **DR. ROSALIND JAMES:**Did you take into account 

### 13 the formulation at all? Is it at this point? Do you ever take 

### 14 into account formulation? 

### 15 **DR. THOMAS STEEGER:**At a screening level, we 

### 16 have available to us formulation toxicity data that are 

### 17 submitted for all different types of formulations. And from 

### 18 that, we get a sense for the extent to which different 

### 19 formulations may be toxic - more toxic than the technical-grade 

### 20 active ingredient. 

### 21 To be candid, those data are on mammals. If 

### 22 we have information for existing chemicals that particular 

### 23 formulations may be more toxic than a technical-grade active 

### 24 ingredient based on open literature data or an incident data, we 

### 25 have the purview to require studies looking specifically at 

### 26 those particular formulations.

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1 **DR. ROSALIND JAMES:**There's a toxicity 

### 2 question, but then also it might affect exposure, like one of 

### 3 the famous cases of microencapsulation where the bees were going 

### 4 after the microcapsules if they were pollen, so they were 

### 5 actually having a higher exposure. So to me, it seems like Tier 

### 6 II might be a place where that came into play, but I wasn't sure 

### 7 if you had considered that. 

### 8 **DR. THOMAS STEEGER:**The microencapsulated 

### 9 products, particularly like Penncap-M that had particular affect 

### 10 on bees, a lot of that information became available to us 

### 11 through incident data. As we'll talk about it in my 

### 12 presentation, incident data serves as a means for the Agency to 

### 13 sort of ground truth its risk assessments with an understanding 

### 14 of how that chemical is performing under actual use conditions. 

### 15 16 So it's a critical need of ours to have access 

### 17 to that information as well as open literature information that 

### 18 would also inform us. But you're correct; there are 

### 19 formulations that you would be able to better glean through a 

### 20 higher tier testing. 

### 21 **DR. DANIEL SCHLENK:**Dr. Potter. 

### 22 **DR. THOMAS POTTER:**I have a quick question. 

### 23 Have you had any studies of this type, the Tier II type or the 

### 24 Tier III type, submitted for your review? 

### 25 **DR. THOMAS STEEGER:**Yes, we have. 

### 26 **DR. THOMAS POTTER:**How did it go?

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1 **DR. THOMAS STEEGER:**I think that the 

### 2 higher tier studies, to date, have been difficult to interpret 

### 3 because they tend to ask or look at very broad questions. One 

### 4 of the things that we're proposing in the White Paper is that as 

### 5 you transition through the various tiers of refinement, the 

### 6 questions that are being asked are intended to address specific 

### 7 uncertainties and that we get away from shotgun approaches to 

### 8 doing field studies to those that are answering very, very 

### 9 refined question. I'm hoping that through that process it'll be 

### 10 much easier to interpret the results of those studies. 

### 11 **DR. DANIEL SCHLENK:**Something to add? 

### 12 **MR. KEITH SAPPINGTON:**I would like to just 

### 13 add that based on my experience in looking at a number of field 

### 14 studies, first of all, in those studies, the typical end-use 

### 15 product was used so we did have a sense of the different 

### 16 formulations. 

### 17 But secondly, and this is, I believe, part of 

### 18 the charge questions, is understanding biological significance, 

### 19 visa-vis, statistical significance because we have difficulties 

### 20 with statistical power. It is important, even more, I think, to 

### 21 have a good sense of what is biologically significant from 

### 22 different endpoints. 

### 23 **DR. DANIEL SCHLENK:**Any other -- Dr. 

### 24 Tarpy. 

### 25 **DR. DAVID TARPY:**Based on Figures 2 and 3 in 

### 26 the White Paper, if a particular compound is shown in Tier I not

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1 to have any concern, then it does not go on to Tier II or III by 

### 2 default, correct? 

### 3 **DR. THOMAS STEEGER:**As Steve Bradbury 

### 4 pointed out earlier, the risk quotients that are calculated at a 

### 5 screening level are not viewed in and of themselves. We take 

### 6 into consideration other lines of evidence that might be 

### 7 available, and those would include open literature studies that 

### 8 might've been conducted; field studies that the registrant may 

### 9 have conducted as part of their testing process, and again, 

### 10 incident data. 

### 11 It's those other lines of evidence that if we 

### 12 feel are in accordance with our understanding of the lack of 

### 13 risk at a screening level, then that would be correct. But if 

### 14 there is information to suggest otherwise, then it would be up 

### 15 to the risk manager to determine whether higher tier levels of 

### 16 refinement would be needed. 

### 17 **DR. DAVID TARPY:**Have there been examples of 

### 18 compounds at Tier I have shown no concern, but tested that Tier 

### 19 II and actually do have effects at that level? 

### 20 **DR. THOMAS STEEGER:**I don't want to cite a 

### 21 particular chemical, but I think that some of the current 

### 22 limitations in the testing process where we don't do -- we 

### 23 haven't done larval toxicity testing as a matter of routine, and 

### 24 compounds that can be completely innocuous to adult bees, but 

### 25 quite detrimental, such as insect growth regulators to the 

### 26 larval stage, might make it through that screen but now we are

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1 proposing a means to address both life stages. 

### 2 **DR. DAVID TARPY:**Thank you. 

### 3 **DR. DANIEL SCHLENK:**Okay. Any other 

### 4 questions before we break? Let's try to be back about five 'til 

### 5 3:00. 2:55. 

### 6 (Brief recess.) 

### 7 **DR. DANIEL SCHLENK:**Okay. Our first 

### 8 presentation is by Joseph DeCant. 

### 9 **MR. JOSEPH DECANT:**Good afternoon. My name 

### 10 is Joseph DeCant, and I'm an ecologist in the Environmental Fate 

### 11 and Effects Division. I'll be talking about the various methods 

### 12 that are available for evaluating the toxicity of pesticides to 

### 13 bees, as outlined in the White Paper. 

### 14 Throughout this talk, I intend to present our 

### 15 studies. The first step, though, will be to provide an overview 

### 16 of how these toxicity studies relate to each other and the risk 

### 17 assessment process, and the overall tiered approached to 

### 18 toxicity evaluation. 

### 19 This talk will also encompass the linkages 

### 20 between the different tiers and introduce the triggers that 

### 21 provide the foundation for moving from one tier to the next 

### 22 within the toxicity-testing framework. Finally, I would like to 

### 23 call your attention to Charge Questions 8 through 12, which this 

### 24 presentation will touch on and to which it will provide context. 

### 25 26 In order to understand the importance of the

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1 toxicity studies themselves and their relation to the risk 

### 2 assessment process, I want to recall Table 1 from the White 

### 3 Paper, regarding the various protection goals. Note, that for 

### 4 each protection goal, there are associated assessment endpoints 

### 5 that will be evaluated in the risk assessment. 

### 6 In addition, there are also associated 

### 7 measurement endpoints that will be evaluated through certain 

### 8 toxicity studies. Therefore, keep in mind that each of the 

### 9 toxicity studies available for the risk assessment should 

### 10 provide one or more measurement endpoints with a clear linkage 

### 11 to an assessment endpoint of concern. 

### 12 As we have already heard in the previous 

### 13 presentations, we have explored methods relating to evaluating 

### 14 exposure, both at the screening level and more refined methods 

### 15 that provide information and actual residues in the crops of 

### 16 interest. 

### 17 The goal of these refinements has been to 

### 18 capture more realistic measures of exposure, albeit at greater 

### 19 expense in terms of the cost of conducting and reviewing such 

### 20 studies. The approach is similar with toxicity studies and 

### 21 their use within the risk assessment process. 

### 22 Consequently, the tiered approach to toxicity 

### 23 testing is meant to provide a means of evaluating the 

### 24 measurement endpoints described in the previous slide through an 

### 25 iterative evaluation process that reflects a progressively more 

### 26 realistic environment and refines the earlier tier evaluations.

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1 For example, at the screening level, toxicity 

### 2 studies are based on the individual in a laboratory; however, as 

### 3 we move through Tiers II and III, we move to studies that 

### 4 evaluate the entire colony and eventually in the field under 

### 5 actual application scenarios for specific labeled uses. 

### 6 For the first year, the U.S. EPA 40 CFR Part 

### 7 158 data requirements currently require an acute contact 

### 8 toxicity test with young adults identified as guideline 

### 9 850.3020. In addition to this test, Canada also requires an 

### 10 acute oral toxicity test with young adult bees. 

### 11 A recent EFED Guidance document, the Interim 

### 12 Guidance on Honey Bee Data Requirements, provided and overview 

### 13 of various studies for use of the screening level and included 

### 14 both the acute contact and oral studies with young adult bees 

### 15 and in an in vitro study for the larval stage of honey bees. 

### 16 Similarly, the proposed approach presented in 

### 17 the White Paper expands upon the current U.S. EPA requirement of 

### 18 the acute contact study to include these two additional oral 

### 19 exposure studies. This expansion is consistent with the 

### 20 USDA-led workshop, as well as two recent publications; one from 

### 21 a 2011 SETAC Pellston Workshop, as well as one from the European 

### 22 Food Safety Authority in 2012. 

### 23 Through the expanded Tier I battery of 

### 24 toxicity tests, we can evaluate the toxicity of a pesticide for 

### 25 both contact and dietary routes of exposure. I also want to 

### 26 mention at this time that these Tier I toxicity tests will be

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1 used to evaluate the stressors of concern to honey bees. 

### 2 Therefore, in some cases, degradates may be identified to be of 

### 3 concern as well. In these cases, toxicity tests will evaluate 

### 4 the parent compound as well as any potentially toxic degradates. 

### 5 6 A U.S. EPA and OECD Guideline evaluate acute 

### 7 contact toxicity to young adult honey bees. The study produces 

### 8 a 48-hour LD50 or a lethal dose at which 50 percent of the test 

### 9 bees dies. The study can be expanded to 96 hours if it is 

### 10 appropriate, as in the case of chemicals that showed delayed 

### 11 toxicity. 

### 12 The most sensitive LD50 values are intended to 

### 13 be used in combination with the appropriate estimated exposure 

### 14 value to derive risk quotients for the Tier I risk assessment. 

### 15 In cases where a pesticide is not applied by foliar spray, the 

### 16 acute contact toxicity data may not be necessary for assessing 

### 17 the risks of a chemical because for soil applications, seed 

### 18 treatments and tree trunk applications, dietary exposure is 

### 19 expected to be the predominant route. 

### 20 Depending on the outcome of these toxicity 

### 21 tests, pesticides are classified as practically non-toxic, 

### 22 moderately toxic, or highly toxic to bees on an acute exposure 

### 23 basis. If the acute contact LD50 is less than 11 micrograms of 

### 24 active ingredient per bee, additional testing may be required in 

### 25 the form of a foliar residue study to determine the duration 

### 26 over which field weathered foliar residues remain toxic to honey

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1 bees. 

### 2 This guideline study, 850.3030, provides an 

### 3 RT25 or the time at which a chemical produces 25 percent or less 

### 4 mortality. The RT25 will not be used to quantitatively evaluate 

### 5 risk, but rather the study will provide another line of evidence 

### 6 related to the contact toxicity of residues on foliages over 

### 7 time. 

### 8 Although not currently required by EPA, Canada 

### 9 and the European Union routinely require acute oral toxicity 

### 10 data for adult worker bees. The acute oral toxicity study is a 

### 11 laboratory test method designed to assess a mortality of young 

### 12 adult worker bees, following a single oral dose of a pesticide. 

### 13 14 A standard OECD Guideline exists for this 

### 15 study. Similar to the acute contact toxicity test required by 

### 16 EPA, the study provides a 48-hour LD50 value and can be extended 

### 17 to include a 96-hour LD50 if mortality is demonstrated to 

### 18 increase between 24 and 48 hours. 

### 19 The proposed Tier I Risk Assessment Methods 

### 20 involves using the 48-hour or 96-hour LD50 generated from this 

### 21 study to derive risk quotients representing risk to adult bees 

### 22 exposed to pesticides through the diet, following foliar spray 

### 23 application, soil application, seed treatments, and tree trunk 

### 24 treatments of pesticides. 

### 25 The previous two slides show the studies that 

### 26 will provide measures of toxicity to adult honey bees. Unlike

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1 with the acute contact in oral toxicity studies for adult worker 

### 2 bees, established regulatory test guidelines for assessing 

### 3 larval toxicity do not exist. Given that larvae may be exposed 

### 4 to pesticides through the diet, the lack of understanding of the 

### 5 toxicity of a chemical on larvae is considered to be a 

### 6 significant data gap. 

### 7 Therefore, efforts are underway to identify 

### 8 critical design elements of an acute oral larval toxicity study 

### 9 that may be used to understand the toxicity of a chemical to 

### 10 this lifestage of worker bees. One option is a field-feeding 

### 11 design that looks at impacts to the brood at the whole hive 

### 12 level. This study is considered a semi-field design and will be 

### 13 discussed later in the presentation. 

### 14 Another option is an in vitro assay based on 

### 15 the method, developed by Aupinel et al., with larval honey bees, 

### 16 which will provide a screening evaluation of toxicity to 

### 17 immature stages. 

### 18 In this method, first instar bee larvae are 

### 19 transferred into 48-well plates and fed a synthetic royal jelly 

### 20 sucrose solution mixture containing known quantities of the test 

### 21 chemical over the 48-hour period. 

### 22 This study can provide an LD50 but it can also 

### 23 potentially provide a no effect concentration, hereafter 

### 24 referred to as NOAEC, if there is sufficient replication. 

### 25 The methodology has been ring tested and a 

### 26 review indicated that additional adjustment in the method may be

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1 needed to adjust potential sources of variability, including 

### 2 colony origin of the brood, season and larval heterogenedia 

### 3 grafting. It is also important to note that the study design 

### 4 has a number of uncertainties associated with it, including 

### 5 laboratory conditions in which the larvae must survive outside 

### 6 of the hive environment. 

### 7 In addition, the larvae are directly fed an 

### 8 artificial mixture of royal jelly and sucrose solution, which is 

### 9 different from what the larvae would be fed, after processing, 

### 10 by adult nurse bees in the hive. 

### 11 The studies that I have discussed so far are 

### 12 meant to evaluate acute toxicity. Chronic laboratory-based 

### 13 toxicity testing has not been routinely required on individual 

### 14 bees in the U.S.; rather information on the potential chronic 

### 15 toxicity of pesticides to bees has historically been discerned 

### 16 from whole colony studies conducted under the semi-field or full 

### 17 field conditions. 

### 18 The proposed Tier I risk assessment method for 

### 19 bees includes quantification of the toxicity of larvae and adult 

### 20 worker bees following chronic exposures to pesticides; however, 

### 21 no formal guidelines have been developed, to date, for 

### 22 conducting chronic toxicity tests with either adult or larval 

### 23 bees. 

### 24 Efforts are underway to identify design 

### 25 elements of chronic larval and adult toxicity studies that may 

### 26 be used to understand the toxicity of a chemical to worker bees

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1 and test protocols already exist for these types of studies. A 

### 2 10-day oral chronic toxicity test with young adults would be 

### 3 used to quantitatively evaluate risk to adults. 

### 4 The USDA technical working group report noted 

### 5 that this type of chronic toxicity test with individual adult 

### 6 bees has already been conducted to support registrations in the 

### 7 EU. These tests consist of collecting newly emerged juvenile 

### 8 bees from the comb and placing 50 of these bees per cage. 

### 9 A sufficient number of replicates are run, 

### 10 along with suitable negative and reference controls to conduct 

### 11 hypothesis testing to establish a NOAEC and LOAEC. The bees are 

### 12 fed a sucrose solution; however, no protein source, such as 

### 13 pollen, is provided. 

### 14 Exposure is through spikes made to the sucrose 

### 15 solution and cages of bees are provided 10 milliliters of test 

### 16 solution every other day. Bees are observed for a total of 10 

### 17 days, and mortality is a primary measurement endpoint. The in 

### 18 vitro chronic larval toxicity study is similar in design to the 

### 19 previously mentioned acute larval toxicity study, though with a 

### 20 longer duration of feeding and monitoring. 

### 21 It should also deliver in NOAEC as well when 

### 22 monitoring of toxic effects is extended to seven days; however, 

### 23 at this point, the chronic adult and larval study methods 

### 24 require further evaluation prior to use in a regulatory setting. 

### 25 Another approach is to derive a no effect 

### 26 concentration from an acute toxicity study; however, because it

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1 is a regression-based study targeted to ascertain the LD50, the 

### 2 dosing regime is not intended to assess the no effect level. 

### 3 Consequently, there is uncertainty as to the actual 

### 4 concentration where no effects occur. 

### 5 Hypothesis-based testing in chronic studies is 

### 6 more appropriate as the dose election targets the low 

### 7 concentrations around the no effect level. Additional data are 

### 8 often available that may be useful in characterizing the effects 

### 9 of a pesticide in bees. 

### 10 The first major bullet notes scientific 

### 11 literature which often includes toxicity data involving bees. 

### 12 Acute toxicity data for honey bees may be available in 

### 13 scientifically valid studies from the open literature, for 

### 14 example, peer-reviewed journal articles. But these studies from 

### 15 the open literature must be sufficiently robust, based on EFED 

### 16 open literature review guides. 

### 17 In some cases, these values may be used to 

### 18 derive risk quotients when these endpoints are lower than those 

### 19 found in registrant-submitted studies. 

### 20 The second major bullet point relates to data 

### 21 on non-target arthropods, other than bees. These data are not 

### 22 used to derive risk quotients, but are considered in the effects 

### 23 characterization. For example, data are often available from 

### 24 registrant-submitted acute toxicity studies involving other 

### 25 non-target arthropods, such as the green lacewing and parasitoid 

### 26 wasps.

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1 The final major bullet point addresses reports 

### 2 of honey bee mortality events -- that is, bee kill incident 

### 3 reports -- following pesticide exposures. These incident 

### 4 reports may be useful in characterizing the available risk 

### 5 estimates from laboratory toxicity data. 

### 6 Although the proposed measurement endpoints 

### 7 for acute and chronic exposures are based on mortality, there 

### 8 are an increasing number of sublethal endpoints reported in Apis 

### 9 and non-Apis toxicity studies, which may be conceptually 

### 10 relevant to the proposed assessment endpoints. 

### 11 Sublethal effects therefore may also provide 

### 12 additional lines of evidence in the risk assessment. These 

### 13 effects are those that do not directly cause the death of an 

### 14 individual organism and are listed in the slide. 

### 15 The acute and chronic studies may provide 

### 16 information on sublethal effects, but they were designed to 

### 17 evaluate mortality and not address concentrations that would 

### 18 elicit sublethal effects. This is the reason that the EFSA 

### 19 Scientific Opinion of pollinator risk assessment concluded that 

### 20 the current battery of toxicity studies are not adequate at 

### 21 addressing sublethal effects. It recommended that sublethal 

### 22 effects are important indicators of bee health and vitality and 

### 23 must be accounted for in the risk assessment process. 

### 24 These endpoints also represent a challenge in 

### 25 that in most cases, they do not provide a direct connection to 

### 26 our assessment endpoints. Before using these endpoints to

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1 derive risk quotients, the relevancy of these sublethal 

### 2 measurement endpoints for evaluating assessment endpoints needs 

### 3 to be established through quantitative links to colony strength 

### 4 and survival. 

### 5 As the last bullet notes, although such 

### 6 endpoints may not be used quantitatively to estimate potential 

### 7 effects on assessment endpoints, studies on sublethal effects 

### 8 could potentially be used qualitatively to describe effects 

### 9 which may not have clearly established relationships to colony 

### 10 level effects or for explaining mechanisms by which colony level 

### 11 effects might occur. 

### 12 Consequently, these sublethal effects will be 

### 13 used to qualitatively describe risk in the assessment until 

### 14 further research has been conducted that establishes a link 

### 15 between these effects and assessment endpoints. 

### 16 This slide provides a representation as to how 

### 17 the data will be used in the risk assessment. The endpoints for 

### 18 quantitative use, which are depicted in the green box, will be 

### 19 combined with either model estimates of exposure or measured 

### 20 residue levels in order to develop risk quotients. 

### 21 The information contained in the sources in 

### 22 the red box will be used qualitatively to further describe the 

### 23 risk that the risk assessor is quantified. This risk analysis 

### 24 will then be related to the assessment endpoints of concern, 

### 25 highlighted in the blue box, and will be used to inform the risk 

### 26 manager, regarding the screening level of results of the use of

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1 a particular pesticide. 

### 2 At the Tier I screening level, effects data 

### 3 may primarily be based on laboratory studies, while exposure 

### 4 estimates may be based on models and/or conservative default 

### 5 values, as well as more refined measures of exposure from field 

### 6 monitoring studies. 

### 7 These studies are evaluated using relatively 

### 8 conservative assumptions to provide high-end estimates of 

### 9 potential risk such that if a chemical passes a screen, the 

### 10 presumption of low risk to non-target organisms is considered, 

### 11 in most cases, protected. 

### 12 As part of the tier process, there's a 

### 13 progression from the left box to the right box in that higher 

### 14 tiered studies are intended to reflect increasing levels of 

### 15 realism, albeit from a less controlled testing environment, in 

### 16 terms of how organisms may be exposed and the nature of the 

### 17 effects which results from such exposure. 

### 18 Also, as part of the tier testing process, 

### 19 higher tiered refinements shift the focus from exposure and 

### 20 effects on individual bees to that of the intact colony. The 

### 21 social nature of honey bees requires extensive interaction and 

### 22 communication among various castes of bees for successful 

### 23 development and propagation of colonies. 

### 24 As a result, numerous linkages exist between 

### 25 pesticide exposure and effects on individual bees and their 

### 26 collective impact on honey bee colony health, which are not

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1 readily assessed in the laboratory. 

### 2 Additionally, the Tier II semi-field studies 

### 3 provide a means of assessing both exposure and effects that are 

### 4 less conservative and more realistic than the methods used in 

### 5 laboratory testing. It is important to note that Tier II 

### 6 studies address the limitation of the Tier I screening level 

### 7 related to sublethal effects, chronic exposure and potential 

### 8 degradate toxicity by integrating these factors into the colony 

### 9 level measurement endpoints. 

### 10 Thus, Tier II studies may be conducted if 

### 11 concerns remain from Tier I levels of the risk assessment and 

### 12 further information is needed for a more refined estimate of 

### 13 potential risk to honey bees. Ultimately, the transition from 

### 14 Tier I to Tier II is based on more than just the relationship 

### 15 between exposure and toxicity. 

### 16 In the case of a chemical that either passes 

### 17 or fails a screen, the risk manager has a number of different 

### 18 types of information available, including the previously 

### 19 described toxicity and exposure data, other lines of evidence, 

### 20 such as incidents, uncertainties related to the characteristics 

### 21 of the chemical itself and whether the Tier I analysis has 

### 22 substantial gaps and knowledge, potential mitigation options, 

### 23 and finally, the option to off-label specific uses. 

### 24 Consequently, Tier II studies are based on the 

### 25 needs of the risk manager and are based on information of the 

### 26 impact of the pesticide, as identified, in lower tier studies.

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1 Unlike the screening level studies, Tier II 

### 2 studies with bees are not typically conducted in the laboratory, 

### 3 but rather focused on relatively controlled field conditions, 

### 4 referred to as semi-field. In these studies, the movement of 

### 5 the bees to and from the colony is restricted through the use of 

### 6 enclosures containing crops treated with the pesticide, or 

### 7 feeding bees a pesticide-spiked diet. 

### 8 Therefore, a Tier II toxicity study is meant 

### 9 to refine the Tier I assessment by providing a means of 

### 10 evaluating the impacts to the assessment endpoints at the whole 

### 11 hive level. Compared to laboratory studies where exposure is to 

### 12 an individual organism and typically through a single route, 

### 13 such as contact or oral exposure, semi-field studies can provide 

### 14 clear lines of evidence for linking multiple routes of exposure 

### 15 to adverse ecological effects at the colony level. 

### 16 Semi-field studies are also useful for 

### 17 determining the extent to which effects on invidivudal bees 

### 18 identified in Tier I laboratory studies are expressed at the 

### 19 colony level. The decision to move to higher tier testing is 

### 20 also supported by current U.S. EPA 40 CFR Part 158 data 

### 21 requirements for the testing of pesticides. 

### 22 The conditions for the transition to higher 

### 23 tier studies are based on the residual toxicity data that 

### 24 indicate there's the potential for effects other than mortality 

### 25 on the colony, and data that indicate potential chronic, 

### 26 reproductive, or behavioral effects.

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1 Although the current EPA data requirements for 

### 2 pesticides identified in the 40 CFR Part 158 do not specifically 

### 3 include semi-field studies, the Guideline 850.3040 on field 

### 4 testing for pollinators is general and would accommodate such 

### 5 studies. 

### 6 The design of the Tier II semi-field study is 

### 7 based upon the specific issues raised at the initial tiers of 

### 8 the risk assessment. The specific design is flexible and, 

### 9 consequently, can be tailored to suit the needs of the risk 

### 10 assessment. 

### 11 In general, there are two basic designs of the 

### 12 semi-field studies. One employs an enclosure, often called a 

### 13 tunnel, as seen in the top picture of the slide, and can find 

### 14 small nucleus hives to ensure that the bees forage on a treated 

### 15 crop. Smaller hives are used in this type of study in order to 

### 16 prevent the colony from overwhelming the forage base within the 

### 17 enclosures. 

### 18 The other design allows bees to freely move, 

### 19 but provides artificial food sources, as seen by the lower 

### 20 picture with pollen cake, spiked with a pesticide of interest. 

### 21 Although the feeding study designs do not involve enclosures and 

### 22 bees are allowed to forage freely, the studies are considered 

### 23 semi-field since test organisms are provided pesticide-spiked 

### 24 food, which does not occur in full field studies. Both designs 

### 25 offer relatively controlled conditions in comparison to Tier III 

### 26 full field studies, yet enable whole hive evaluations to be on

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1 Tier I screening laboratory studies. 

### 2 As is evident, there are a range of different 

### 3 designs and the actual test structure will depend on the 

### 4 specific questions asked in the risk assessment. There are a 

### 5 number of already established guidelines for the semi-field 

### 6 tunnel study. 

### 7 The typical designs of semi-field studies 

### 8 include a highly pollinator attractive crop or surrogate species 

### 9 such as phacelia that is grown outdoors in an enclosed system 

### 10 with confined exposure. 

### 11 The test crop is grown under good 

### 12 agricultural practices using the maximum application rate with 

### 13 the minimum application interval. The test crop provides the 

### 14 source of pollen and nectar. The test design employs a tunnel 

### 15 in which small nucleus hives containing about 5,000 worker bees, 

### 16 brood in all stages, and stores of pollen and nectar are placed. 

### 17 A test crop is used at or near full bloom that 

### 18 is representative of high-end exposure condition for the given 

### 19 type of application method and rate. For foliar sprays, the 

### 20 hives are introduced in the tunnel two to three days before 

### 21 pesticide application and hive condition is assessed while the 

### 22 bees acclimate to the enclosure. 

### 23 Pesticides can then be applied at peak bee 

### 24 foraging activity to maximize exposure to foraging bees. Hives 

### 25 are kept in the tunnel for a period of seven to ten days, after 

### 26 the first pesticide application and allowed to forage on the

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1 treated crop. The hives are then removed from the tunnel and 

### 2 moved to an area of minimal pesticide use to forage on untreated 

### 3 areas with continued monitoring, which is usually up to a total 

### 4 study period of 28 days but sometimes longer. 

### 5 A reference chemical is also frequently tested 

### 6 to demonstrate the application method used that actually results 

### 7 in exposure to bees, as well as the ability of the study to 

### 8 detect and quantify effects. Reference chemical selection is 

### 9 dependent on the expected mode of action of the pesticide under 

### 10 consideration, which would be a chemical with known toxicity to 

### 11 the hive. 

### 12 For pesticides that are expected to directly 

### 13 impact brood development, an insect growth regulator, such as 

### 14 fenoxycarb, is typically used. For pesticides that are expected 

### 15 to elicit effects directly on adults and indirectly on brood, 

### 16 through rapid knockdown, a reference chemical that acts 

### 17 similarly, such as dimethoate, is often selected. 

### 18 In addition to the monitoring of toxic 

### 19 effects, residues in pollen and nectar on the flowering crop can 

### 20 also be provided in order to assess the levels of exposures to 

### 21 the honey bee colonies. 

### 22 As the last bullet highlights, it is important 

### 23 to note that this design can be used to evaluate each of the 

### 24 application methods, depending on when the tunnels are in place. 

### 25 Moving to the feeding designs, one design 

### 26 typically addresses brood toxicity. The study is based on a

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1 method by Oomen, et al. in 1992, which uses hives that can 

### 2 freely forage in order to test for the possibility of adverse 

### 3 effects on the brood within a honey bee hive. The basic design 

### 4 uses one liter of a sucrose solution that is placed in a feeder 

### 5 near a full-sized honey bee hive. The sucrose solution is 

### 6 either a control without contamination or a solution spiked with 

### 7 a known concentration of a pesticide. 

### 8 The solution is provided as a food source and 

### 9 once the hive has collected all of the solution, weekly 

### 10 measurements are taken on brood development within the hives. 

### 11 In addition to the data on brood development, data on adults and 

### 12 brood mortality estimates can be obtained through counts of bees 

### 13 collected in dead bee traps, placed at the entrance of the 

### 14 colony. 

### 15 The test chemical concentrations are not 

### 16 necessarily meant to be reflective of an environmentally 

### 17 relevant concentrations, but rather may be exaggerated in order 

### 18 to screen chemicals for possible impacts to the brood when in a 

### 19 whole hive situation. 

### 20 The EPA has also explored the modification of 

### 21 the feeding design to expand it to adults as well as brood 

### 22 toxicity, to incorporate multiple dietary pathways of exposure 

### 23 and to increase the duration of the study if necessary. It 

### 24 should be noted that these refinements have not been well 

### 25 vetted, but a number of field feeding designs are increasingly 

### 26 found in the open literature, reflecting the potential for this

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1 type of experiment to address uncertainties related to dietary 

### 2 exposure. 

### 3 For this type of study, spiked food, as in 

### 4 pollen patties, sucrose solution or both, can be fed ad libitum 

### 5 to full size honey bee hives of at least 10,000 bees over a 

### 6 specified duration of time and during a specified time of the 

### 7 year. Different concentrations of the chemical and food are 

### 8 used with the intent of identifying a NOAEC and LOAEC. 

### 9 The treatment levels are either selected from 

### 10 concentrations of concern that have been identified in the body 

### 11 of literature or by environmentally relevant concentrations 

### 12 found in targeted monitoring studies for residues in pollen and 

### 13 nectar. Therefore, the design should be based on 

### 14 environmentally relevant concentrations in an attempt to 

### 15 ascertain where the no effect level is found, relative to these 

### 16 concentrations. 

### 17 The design does not employ tunnels, so the 

### 18 bees are allowed to forage freely. As the second bullet notes, 

### 19 while the potential exists for bees to forage in areas where 

### 20 other pesticides may be in use, test site selection is intended 

### 21 to minimize the potential for such exposure. Consequently, the 

### 22 test hives are placed in an area of low pesticide use in order 

### 23 to minimize any exposure to other pesticides. 

### 24 Related to the third bullet, during a field 

### 25 feeding study, exposure can be controlled to a greater extent 

### 26 than a full field study through the use of pollen traps, to

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1 limit incoming pollen and encourage feeding on the treated food 

### 2 source. As previously mentioned, the enclosures used in the 

### 3 typical semi-field design introduced stress to the colony and 

### 4 effective brood performance on the colonies in as little as 

### 5 seven to ten days. 

### 6 Moving to the fourth bullet, the open field 

### 7 feeding design allows a colony to freely forage, thereby 

### 8 eliminating the stress of the enclosure. Furthermore, as the 

### 9 fifth bullet states, the study hives can be large enough to 

### 10 survive through overwintering, unlike the smaller nucleus hives 

### 11 typically used in the tunnel studies. 

### 12 The use of larger colonies enables a study to 

### 13 proceed over a longer duration than other semi-field studies in 

### 14 an enclosure. With this flexibility, the full field feeding 

### 15 design can be used to evaluate a colony's response to either 

### 16 short-term or a long-term exposure condition to evaluate 

### 17 overwintering performance of the hives. 

### 18 Consequently, this study can address the 

### 19 uncertainties identified at the lower tier levels or even at the 

### 20 initial stages of the Tier II toxicity assessment with other 

### 21 semi-field studies, specifically when oral exposures is the 

### 22 pathway of concern. 

### 23 Finally, as the last bullet states, the 

### 24 measurement endpoints can be taken at specified time periods to 

### 25 evaluate the hive at the most relevant time points, relative to 

### 26 the nature and concern of the chemical.

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1 The results of this study are intended to 

### 2 reflect to the extent to which specific effects may occur in 

### 3 response to various pesticide levels in pollen and nectar when 

### 4 compared to measured residues from targeted monitoring studies. 

### 5 Assuming that targeted and monitoring studies 

### 6 are available to design the field feeding study or are available 

### 7 following the completion of the feeding study, these data would 

### 8 provide a measure of the level of residues in pollen and nectar 

### 9 from various crops. This feeding study could then provide a 

### 10 context for these residue data at the whole hive level. 

### 11 Given the assumption that exposure to 

### 12 concentrations in the pollen and nectar are equivalent to 

### 13 exposure from concentrations in the sucrose solution and/or 

### 14 pollen patties, the study would reveal to what extent the risk 

### 15 assessor could anticipate effects at the residue levels in the 

### 16 different crops by comparing the concentrations that produce an 

### 17 effect and measured concentrations in pollen and nectar. 

### 18 The comparisons between the no effect level 

### 19 and the residue levels are not quantitative comparisons to be 

### 20 used as triggers, but rather a means of understanding the 

### 21 potential impacts on the colony, given realistic residue levels 

### 22 in pollen and/or nectar from various crops. 

### 23 The study results could also further describe 

### 24 the nature of these impacts to the risk manager to consider with 

### 25 other lines of evidence. The measurement endpoints are 

### 26 determined by the protection goals and remaining uncertainties

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1 from the lower tiers in coordination with the risk manager. 

### 2 Measurement endpoints include adult bee 

### 3 mortality, brood condition, hive strength, queen health, and 

### 4 hive resources which are measured at various points over the 

### 5 duration of the study. 

### 6 The endpoints should also be specific to the 

### 7 uncertainties identified earlier in the tiered process as they 

### 8 relate to the assessment endpoints, so as to minimize the number 

### 9 of endpoints to be collected because continued opening of the 

### 10 hives can add stress to the colony. 

### 11 Another major strength of these semi-field 

### 12 studies is that they integrate effects from toxic degradates, 

### 13 chronic exposure and sublethal effects. Two prominent 

### 14 uncertainties at the Tier I screening level are that the 

### 15 laboratory studies either are inadequate to screen for various 

### 16 types of sublethal effects and there are no currently 

### 17 established protocols to evaluate chronic toxicity. 

### 18 The endpoints measured in the semi-field 

### 19 studies integrate each of the previously mentioned factors and 

### 20 that have toxic degradates, chronic exposure of sublethal 

### 21 effects impact these measurement endpoints which are also tied 

### 22 to assessment endpoints of concern and the study reflects the 

### 23 sum of both these lethal and sublethal effects. 

### 24 In addition to the strength of the semi-field 

### 25 studies, it is also important to note that there are limitations 

### 26 to these study designs as well. Since the studies are conducted

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1 outside of a laboratory environment, feeding and tunnel designs 

### 2 are subject to the changes of weather, as noted in the OECD 

### 3 Guidance for Semi-Field Studies, the test cannot be performed 

### 4 under adverse weather conditions. In addition, there is a high 

### 5 degree of variability in the measurement endpoints, not only 

### 6 between hives, but within the same hive over time as well. 

### 7 Related specifically to the tunnel design 

### 8 because bees are not allowed to forage freely over large areas, 

### 9 the quantity and quality of food they are able to gather may be 

### 10 compromised, compared to open field conditions. Experience with 

### 11 these studies indicates that confinement stress on foraging 

### 12 bees, among other factors, often leads to adverse effects on 

### 13 egg, larval, and pupil abundance, with increasing duration of 

### 14 confinement in the tunnels. 

### 15 As a result, hives can only be kept in tunnels 

### 16 for a relatively short period of time that is seven to ten days, 

### 17 to ensure adequate condition of controls for comparison. Even 

### 18 during this time declines in control hive condition are commonly 

### 19 encountered. 

### 20 Furthermore, effects at other critical time 

### 21 periods of hive development, such as overwintering, are not 

### 22 addressed. Although such effects could be assessed if the post 

### 23 tunnel observation period were extended and colonies were of 

### 24 sufficient strength to support overwintering. 

### 25 For the feeding studies, the bees are allowed 

### 26 to freely forage, so consumption of the spiked food provided to

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1 the colony depends on a range of factors that may not be 

### 2 constant for all experimental conditions. For example, if 

### 3 alternative forage is ample, then the bees may use less spiked 

### 4 sucrose solution or spiked pollen then when outside forage 

### 5 sources are scarce. Thus, this type of study, which does not 

### 6 use an enclosure, is subject to numerous sources of variability 

### 7 at the field level, including weather, disease, parasites and 

### 8 exposure to other pesticides present in the environment. 

### 9 On the second bullet, the feeding aspect is 

### 10 artificial and may not accurately mimic typical foraging 

### 11 behavior by honey bees, or the stability of the test material in 

### 12 such matrices under natural conditions. 

### 13 The third bullet, the foragers only need to 

### 14 move minimal distances to the food source, therefore, use of an 

### 15 internal spiked food source can confound efforts to integrate 

### 16 impacts on foraging behavior when foragers feed on a treated 

### 17 crop. 

### 18 The fourth bullet, the performance of the 

### 19 colonies has not been evaluated in relation to long-term 

### 20 continual use of the artificial food source. And the final 

### 21 bullet notes that as a study increases in duration, it also 

### 22 increases in complexity and potentially confounding factors, 

### 23 such as the interventions required to enable a colony to 

### 24 overwinter. 

### 25 It is essential to highlight the importance of 

### 26 establishing the statistical significance necessary to

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1 adequately evaluate a desired level of protection based on 

### 2 biological significance. The issue of statistical and 

### 3 biological significance relates to both semi-field and full 

### 4 field studies; the latter of which will be explored later in the 

### 5 presentation. 

### 6 Both of these types of studies seek to 

### 7 evaluate the potential for an effect, relative to the control 

### 8 group at a specified application rate or concentration within a 

### 9 supplied food source. In some cases, we are able to evaluate 

### 10 time trends and hive performance, though these are not routinely 

### 11 reported. Yet there is a high level of variability typically 

### 12 associated with the measurement endpoints in both types of 

### 13 studies, where variability may be so high that it is difficult 

### 14 to isolate any effect. 

### 15 While 10 percent mortality has been 

### 16 established as a level of protection at the screening level, a 

### 17 level of biological significance has not yet been established 

### 18 that links measurement endpoints at either the semi-field or the 

### 19 full field level to the assessment endpoints. Furthermore, 

### 20 because of compensatory mechanisms within the colony, apparent 

### 21 effects may only be transitory. 

### 22 Well designed semi-field studies, despite 

### 23 their uncertainties, should be able to inform the risk manager 

### 24 by providing clear lines of evidence linking multiple routes of 

### 25 exposure to adverse ecological effects and provide for a more 

### 26 robust characterization of the risk related to the assessment

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1 endpoints. 

### 2 Semi-field studies assess both exposure and 

### 3 effects at the colony level that are less conservative and more 

### 4 realistic than laboratory assays on the individual bees. These 

### 5 studies also integrate effects from toxic degradates, sublethal 

### 6 effects and chronic exposure. 

### 7 Furthermore, the Tier II feeding study for 

### 8 adults and brood should provide a no effect concentration that 

### 9 can then be compared to residue levels in various crops 

### 10 identified in targeted residue studies. 

### 11 Unlike the Tier I toxicity studies that are 

### 12 used to calculate risk quotients, the use of the Tier II studies 

### 13 will be used to qualitatively evaluate risk and to help 

### 14 characterize the quantitative risk estimates derived from 

### 15 laboratory studies on the individual at the first tier. 

### 16 Consequently, semi-field studies will help 

### 17 inform the risk manager regarding any need for mitigation 

### 18 options or options for further refinement. When uncertainties 

### 19 remain from the Tier II assessment or risk cannot be mitigated, 

### 20 the risk manager may decide that additional data are necessary 

### 21 in the form of Tier III full field studies. 

### 22 The transition to Tier III is based on 

### 23 addressing remaining uncertainties at the previous tiers. This 

### 24 slide highlights some of the major uncertainties inherent to the 

### 25 semi-field study designs and how the Tier III full field study 

### 26 may address those design limitations. In the case of the tunnel

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1 study, a full field study does not employ tunnels and therefore 

### 2 eliminates the stress of the tunnel on the various aspects of 

### 3 colony functioning, as well as limitations in the size of the 

### 4 colonies and the duration of the study. 

### 5 Related to the feeding designs, a Tier III 

### 6 full field study allows hives to naturally forage and it also 

### 7 allows a study to integrate any impacts on the foragers that 

### 8 visit the treated crop or consequences to the hive based on 

### 9 foraging on the treated crop. 

### 10 In summary, the full field study overcomes the 

### 11 major limitations of the stress placed on the colonies from the 

### 12 enclosure in the tunnel design and the artificial exposure 

### 13 pathway in the feeding designs. 

### 14 A Tier III study should further refine any 

### 15 uncertainties identified by the Tier II assessment, related to 

### 16 the assessment endpoints. The Tier III study is a full field 

### 17 design that provides the highest level of refinement of the 

### 18 tunnel and feeding study designs. 

### 19 Consequently, Tier III effects assessments are 

### 20 intended to represent highly refined studies that address 

### 21 specific uncertainties and/or risks identified in lower tiered 

### 22 studies. 

### 23 Similar to the transition from Tier I to Tier 

### 24 II, the risk manager will consider the availability of multiple 

### 25 sources of information. The need to transition to Tier III is 

### 26 therefore not an automatic requirement, but rather, based on

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1 multiple lines of evidence. 

### 2 A field study serves as a means of addressing 

### 3 uncertainties raised in lower tier studies conducted either in 

### 4 the laboratory or under semi-field conditions and can be useful 

### 5 in examining the effects of pesticides with extended residual 

### 6 toxicity. 

### 7 However, a full field study is intended to 

### 8 represent realistic application conditions, whereby the test 

### 9 substance is applied to a specific crop in which bees are 

### 10 foraging freely, without the use of an enclosure, and whereby 

### 11 test colonies are primarily exposed through residues carry back 

### 12 to the hive by the forager bees. 

### 13 Consequently, a full field study is also the 

### 14 most resource-intensive, relative to studies used for the Tier I 

### 15 and II assessments, to conduct in terms of both time and money. 

### 16 These studies can also be the most difficult to interpret given 

### 17 the level of associated variability. Therefore, they can 

### 18 represent a significant investment to regulatory agencies in 

### 19 terms of the resources that are required to review these 

### 20 studies, but these studies may also provide contacts relative to 

### 21 other lines of evidence. 

### 22 By serving as a study to refine the 

### 23 uncertainties and effects identified in the previous tiers, the 

### 24 field study is also meant to test for the potential of an effect 

### 25 and not the mechanistic cause of an effect. Furthermore, the 

### 26 Tier III field study should have a focused risk hypothesis, that

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1 is, a risk hypothesis informed by and built upon the previous 

### 2 tiers. 

### 3 There are a number of guidance documents 

### 4 related to the conduct of a field study. Currently, the EPA has 

### 5 guidance for conducting the pollinator field study. 

### 6 Similarly, the EPPO Guidance also addresses 

### 7 pollinator field study design. Regarding the first major 

### 8 bullet, the EPA field pollinator guidance does not provide much 

### 9 detail, but rather, provides flexibility for designing a study 

### 10 based on the needs of the risk assessment. 

### 11 The second major bullet mentions Guideline 

### 12 850.2500. Although intended as guidance for conducting field 

### 13 studies with mammals and birds, this EPA Guidance titled, "Field 

### 14 Testing for Terrestrial Wildlife" provides very useful 

### 15 information on study design elements to consider for field 

### 16 testing. 

### 17 This guidance addresses the basic purpose of a 

### 18 field study, geographic area and selection of study sites, 

### 19 sample size, and the test conditions of the study. EPPO 

### 20 Guidance provides more specific detail that can be used to 

### 21 design a field pollinator study with honey bees in terms of 

### 22 experimental conditions, such as selection of the crop, 

### 23 placement of the colonies, relative to the treated field, size 

### 24 of the colonies and plot size. 

### 25 Also, application of treatments, such as use 

### 26 of controls, timing of application and rates; and finally, mode

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1 of assessment, such as timing and frequency of assessments. 

### 2 Similar to Tier II studies, full field studies 

### 3 are flexible in terms of their specific design, as they are 

### 4 based on uncertainties highlighted in the previous tiers, as 

### 5 well as the nature of the pesticide. Consequently, registrants 

### 6 should submit protocols for the field study prior to study 

### 7 initiation in order to identify the key design elements based on 

### 8 specific risk management concerns. In general, though, there 

### 9 are design elements common to most field studies. 

### 10 The second bullet notes that a field study for 

### 11 honey bees, in general, should be conducted under actual use 

### 12 conditions in which a typical end-use product is used at its 

### 13 maximum application rate, frequency and method, as described in 

### 14 the label for a specific crop. The design should reflect a 

### 15 realistic foraging environment and exposure system in the field 

### 16 where the pesticide is to be applied. 

### 17 The third bullet notes that the study is 

### 18 conducted in a crop grown outdoors without enclosures. 

### 19 Consequently, the primary exposure route is to foragers that are 

### 20 allowed to freely fly and forage. 

### 21 The fourth bullet notes that the crop is 

### 22 subject to good agricultural practices. And the fifth bullet 

### 23 mentions the size of the hives, each with a minimum of 

### 24 approximately 10,000 bees which are placed within or at the edge 

### 25 of treated or controlled flowering plots and monitored for a 

### 26 specified period of time, depending on the concerns of the

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1 chemical, as in the case of a systemic compound that is 

### 2 persistent in high food stores. 

### 3 Finally, the last bullet deals with the 

### 4 duration of the study. The evaluation of the hive should 

### 5 ideally proceed for a minimum of 50 days, based on the lifecycle 

### 6 of the honey bee within the colony. Because hives may differ in 

### 7 size, colonies should be distributed as equitably as possible 

### 8 between treatments. The colonies should be in position 

### 9 approximately two to three days before the trial in order to 

### 10 acclimate to the study site. 

### 11 If the chemical is a foliar spray, treatments 

### 12 should be applied when the test crop is in full flower during 

### 13 the daytime when bees are demonstrated to be actively foraging 

### 14 on the test crop. Hives should be queen ripe; that is to say 

### 15 have a single functional queen. Use sister queens that is a 

### 16 common maternal source and be of similar strength at the start 

### 17 of the study. 

### 18 Equipment should be free of other chemicals 

### 19 and the hives should have low incidences of disease and 

### 20 parasites, though it is nearly impossible to use hives complete 

### 21 free of disease and pests. Unlike Tier II studies, there are 

### 22 currently no pesticides identified as an adequate positive 

### 23 control at the full field Tier III level. 

### 24 The range of endpoints that may be measured, 

### 25 as well as the selection of endpoints in the Tier III full field 

### 26 study are similar to those in the Tier II semi-field study. As

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1 mentioned in the Tier II section, the endpoints should also be 

### 2 specific to the uncertainties identified earlier in the tier 

### 3 process as they relate to the assessment endpoint so as to 

### 4 minimize the number of endpoints to be collected and the 

### 5 continued opening of the hives. 

### 6 Also similar to the Tier II study designs, 

### 7 Tier III studies integrate toxic degradates, chronic exposure 

### 8 and sublethal effects into the measurement endpoints. A major 

### 9 advantage of the Tier III field study is that the duration of 

### 10 the study with possible overwintering and the consequent timing 

### 11 of measurements are flexible and could be suited to the 

### 12 properties of the chemical under actual use conditions. 

### 13 In order to adequately interpret a field 

### 14 study, it would be helpful for the study to properly evaluate 

### 15 exposure. It can be difficult to evaluate the effects of the 

### 16 test chemical within a full field study because the design 

### 17 allows for bees to forage freely and to obtain food both onsite 

### 18 and off the treated site. 

### 19 In studies with limited bloom density on the 

### 20 treated crop and extensive alternative forage in the vicinity, 

### 21 bees may actually forage very little in the test plot and this 

### 22 may lead to an underestimation of potential effects on the hive 

### 23 due to reduced exposure or alternatively to an inability to 

### 24 detect treatment effects due to controlled contamination 

### 25 There are a number of ways of measuring the 

### 26 degree to which the colony is exposed to the test substance.

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1 First, pollen traps are fixed at the entrances of the hives, 

### 2 force pollen off of the legs of a percentage of foragers when 

### 3 they return to the hive. This pollen can then be analyzed for 

### 4 residues. 

### 5 Second, freshly stored nectar and pollen or 

### 6 stored bee bread and honey can be measured in the hives to 

### 7 determine the level of residues that the bees are storing for 

### 8 consumption or processing prior to feeding of the queen and 

### 9 brood. 

### 10 Measures of residues in the blossoms would 

### 11 provide a comparison of stored concentrations with those 

### 12 collected from the blossoms. When pollen is collected from the 

### 13 pollen trap, the study may analyze floral origin and percent 

### 14 composition of the pollen. Some pollen from the flowers may 

### 15 also be collected by the bees and mixed with the nectar when it 

### 16 is stored. So in addition to sampling of residues and freshly 

### 17 stored nectar, pollen could potentially be obtained from within 

### 18 this collected nectar to determine the floral origin of the 

### 19 nectar as well. 

### 20 Finally, qualitative measures of foraging 

### 21 activity, potential quantitative measures of the return of 

### 22 forager bees and quantitative measure of bloom density can 

### 23 provide information on the potential attractiveness of the crop 

### 24 of interest. 

### 25 The Tier III full field study is complex and 

### 26 difficult to carry out. So there are a number of design

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1 challenges that we face and need to address. Consequently, we 

### 2 are requesting feedback on the design challenges of the field 

### 3 studies and our suggestions for improving these designs that 

### 4 will be covered later in the presentation. 

### 5 Concerning the design challenges, honey bees 

### 6 have an extensive foraging range. Given an alternative to 

### 7 forage, honey bees may minimally forage on the test fields as 

### 8 there is no means of controlling where honey bees forage. In 

### 9 addition, uncontaminated food reserves collected prior to study 

### 10 initiation may result in delayed exposure of the colonies to the 

### 11 test substance, as these previously collected food reserves may 

### 12 be used first. 

### 13 The weather can also affect the ability of 

### 14 bees to forage, where similar to the Tier II studies, adverse 

### 15 weather may prevent honey bees from foraging on a test crop. 

### 16 Finally, to date, typical plot sizes in field studies are 

### 17 minimal, relative to both the representative acreage of some 

### 18 crops in North America and the potential foraging radius of the 

### 19 honey bee. 

### 20 This slide represents the range of plot sizes 

### 21 for typical field studies ranging in sizes from two and a half 

### 22 to 13 acres, which equates to .004 to .02 square miles. 

### 23 Relative to the maximum foraging area of the honey bee colony, 

### 24 based on a five-mile foraging radius, this represents .0051 to 

### 25 .025 percent of the potential maximum foraging area. The 

### 26 question then remains: what size a plot would satisfy the needs

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1 of the colony to ensure that it forages to the maximum extent on 

### 2 the treated field; especially considering that replication of 

### 3 the colonies at the within plot level is typically three to four 

### 4 full size hives that can each be up to 50,000 to 60,000 bees. 

### 5 The colony size is another factor to consider. 

### 6 The EFSA Scientific Opinion and pollinator risk assessment also 

### 7 notes several points related to the advantages of different 

### 8 sizes of the hives for the field study. Larger populations may 

### 9 be more efficient honey producers and produce more honey on a 

### 10 per bee basis while using less per bee over the winter. 

### 11 Thus, a larger colony with more foragers and 

### 12 in nest bees could provide a better observation of the effects 

### 13 of the pesticide because of the higher number of bees involved. 

### 14 However, the EFSA Scientific Opinion also notes that smaller 

### 15 colonies may be more sensitive due to reduced resilience to 

### 16 replace foragers with nurse bees in a smaller brood area. 

### 17 The second major bullet notes that hives at 

### 18 the full field level cannot be completely protected from the use 

### 19 of other pesticides, either on the test field itself or adjacent 

### 20 to the test fields. These products may be applied to the same 

### 21 crop as part of grower's standard practice or they may be 

### 22 applied on neighboring fields on which honey bees may forage or 

### 23 from which the pesticides may drift into the test or control 

### 24 plots of a full field study. 

### 25 In the case of persistent pesticides, these 

### 26 chemicals may remain on the test field or in the adjacent field

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1 well after the application. These products may then be 

### 2 collected by honey bees and brought back to the hive, thereby 

### 3 serving as a source of exposure to other chemicals or even the 

### 4 chemical under study. 

### 5 As the third major bullet highlights, the 

### 6 treatment and control fields should also be adequately 

### 7 separated, though logistical constraints associated with 

### 8 replication of sites may make this difficult. 

### 9 For control in treatment plots with inadequate 

### 10 separation where honey bees can forage freely, bees form control 

### 11 plot may forage on treated plots, thereby bring contaminated 

### 12 food into the control hives. Conversely, treatment plot hive 

### 13 bees may forage in control plots as well, thereby decreasing the 

### 14 extent of exposure. This movement of control and treatment plot 

### 15 bees between plots can confound the interpretation of the field 

### 16 study. 

### 17 Parasites and certain diseases can also affect 

### 18 honey bee colonies, as the fourth major bullet states. Some of 

### 19 these can impact the same measurement endpoints as the pesticide 

### 20 of interest. 

### 21 The next major bullet highlights the duration 

### 22 of the study that also presents a challenge, especially when the 

### 23 study is carried through overwintering and attempts to ascertain 

### 24 the effect of a pesticide while minimizing any potential 

### 25 confounding effects of the interventions that are required to 

### 26 enable a honey bee colony to survive overwintering.

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1 The final bullet addresses replication and 

### 2 variability. As previously noted in the section on Tier II 

### 3 studies related to variability and biological significance, the 

### 4 level of variability in the measurement endpoints is relatively 

### 5 high, due to all of the potential confounding factors at the 

### 6 full field level. 

### 7 As a result of these design challenges, we 

### 8 have identified a number of modifications that may enable more 

### 9 robust field designs. The size of the plots should increase to 

### 10 be more reflective of the size of representative crop acreage in 

### 11 North America. The size of the areas should also adequately 

### 12 support the nutritional requirements of the hive for the 

### 13 duration of the study with the specific purpose of maximizing 

### 14 the foraging of the hives on the test crop to provide an upper 

### 15 bound exposure scenario for that crop. 

### 16 It is recognized, however, that the bloom 

### 17 period of the test crop may be short, relative to the overall 

### 18 study period. But the study design must consider that the 

### 19 nutritional requirements of the study colonies are met. 

### 20 A field study should also provide for adequate 

### 21 separation of treated and control plots, however, climatic and 

### 22 landscape variations may preclude greater distances between 

### 23 plots, depending on these variations over varying geographic 

### 24 scales. 

### 25 According to the EFSA Scientific Opinion, the 

### 26 field test sites should use areas with similar environmental

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1 conditions, where possible, with at least four to six kilometers 

### 2 that is 2.5 to 3.7 miles between treated and control plots. 

### 3 Statistical analysis should be conducted for 

### 4 all field studies; therefore, field studies should also 

### 5 determine the power of the test to adequately assess the 

### 6 endpoints relative to the appropriate level of biological 

### 7 significance. An appropriate power analysis should accompany 

### 8 each protocol proposal in order to determine the number of hives 

### 9 necessary for the conduct of the study. 

### 10 In addition, the final report should also 

### 11 contain an appropriate power analysis to determine if the level 

### 12 of statistical power has been achieved. Measures of exposure 

### 13 are essential to the interpretation of the field study. 

### 14 Consequently, it would be helpful for the study to measure 

### 15 residues in incoming pollen through the use of pollen traps 

### 16 and/or nectar, as in sampling of the honey bee stomach; measure 

### 17 residues and store pollen and nectar pre-exposure and at various 

### 18 times, post-exposure; measure bloom intensity and duration of 

### 19 that intensity and identify sources and percent composition of 

### 20 pollen and nectar. 

### 21 The studies should also limit disease and 

### 22 pests using new hive equipment and obtain bees from sources with 

### 23 low pest occurrence. This slide presents other modifications 

### 24 that could help to improve the quality of the field studies that 

### 25 regulatory agencies receive. The duration of the study should 

### 26 match the concern of the chemical. According to the EFSA

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1 Scientific Opinion, the colonies that are used in the experiment 

### 2 should be monitored for a period of time, covering the entire 

### 3 flowering period and last at least two brood cycles. When 

### 4 residues are persistent, monitoring should continue through 

### 5 winter to after the overwintering period. 

### 6 Other recommendations proposed by the EFSA 

### 7 Scientific Opinion, related to field study designs, include the 

### 8 use of a highly attractive crop, quantitative assessment of 

### 9 brood condition, residue analysis of in hive bees and evaluation 

### 10 of the storage of food and their associated residue levels, and 

### 11 in some cases, the removal of food stocks from the hive at study 

### 12 initiation to maximize the exposure to the test substance. 

### 13 Given the previously mentioned modifications 

### 14 to the design of the field study, it is important to note that 

### 15 some of them notably increasing the plot sizes and ensuring 

### 16 adequate replication would likely be difficult to implement, 

### 17 given the logistics of a field study. 

### 18 The purpose of these studies is that they 

### 19 provide important information relative to the risk assessment. 

### 20 Note, that as we move up the tiers to Tier III, we not only 

### 21 increase the realism, but we also increase the variability due 

### 22 to the number of potentially confounding factors as well as the 

### 23 cost, in terms of time, money, and resources. 

### 24 In fact, full field studies have cost a 

### 25 considerable amount of resources, both to conduct and review, 

### 26 without being classified as acceptable, and only providing

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1 minimal gains, in terms of the risk assessment. 

### 2 In these cases, field studies have not been 

### 3 particularly refined, but rather, measured numerous endpoints, 

### 4 whether quantitatively related to the assessment endpoints 

### 5 outlined earlier or not. 

### 6 Furthermore, there is uncertainty in the 

### 7 results, due to factors, such as minimal distance between 

### 8 treated and control plots that allows the forager bees of the 

### 9 two treatment groups to cross forage; the exposure of test 

### 10 colonies to numerous other chemicals that are used as standard 

### 11 grower practice and the lack of adequate replication given high 

### 12 levels of variability and the measurement endpoints. 

### 13 To date, for the studies that we've seen 

### 14 related to the exposure studies, the refined exposure studies, 

### 15 there appears to be less variability in those studies relative 

### 16 to the ones that assess effects to the bees at the whole hive 

### 17 level. 

### 18 The previously mentioned modifications to the 

### 19 Tier III field study should enable a stronger, more robust 

### 20 design. A properly design and implanted field study should 

### 21 therefore provide essential information on actual uses of the 

### 22 pesticide of interest that can inform the risk manager in any 

### 23 additional mitigation options, if available. 

### 24 The field study should also provide a more 

### 25 clear linkage between the measurement endpoints and the 

### 26 assessment endpoints from the actual use of the chemical, as a

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1 field study can be screening or definitive in nature to either 

### 2 determine the presence or absence of an effect or define the 

### 3 magnitude and duration of an effect. 

### 4 The ultimate design of the study will be 

### 5 dictated by the specific uncertainties it is intended to address 

### 6 in order to qualitatively describe the risk, effects and 

### 7 uncertainties identified in the lower tiers. While these 

### 8 studies are conducted in response to concerns identified in 

### 9 lower tier studies, the full field study is ultimately intended 

### 10 to address whether risks estimated using lower tier exposure and 

### 11 effect data occur under actual use conditions. 

### 12 It is also important to note that the field 

### 13 study will not be interpreted in isolation of the other tiers 

### 14 and other information. Rather, we will evaluate and interpret 

### 15 the field study and their risk assessment using all of the data 

### 16 available to the risk assessor, including data from laboratory 

### 17 toxicity studies at the first year, open literature, non-target 

### 18 arthropod studies, sublethal effects, incident reports, other 

### 19 semi-field or full field studies and the residue levels in 

### 20 various crops, based on exposure data. This weight of evidence 

### 21 approach will be further explained in the next presentation. 

### 22 Thank you for your time. We can now take any 

### 23 questions that the panel may have. 

### 24 **DR. DANIEL SCHLENK:**Okay. Dr. Berenbaum. 

### 25 **DR. MAY BERENBAUM:**I may have missed this, 

### 26 but in transitioning from Tier I to Tier II to Tier III, is

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1 there a task that assumption that colony genetics will be 

### 2 controlled for or minimally subspecies? Is there a 

### 3 consideration of consistency in terms of genetic identity from 

### 4 one tier to the next? 

### 5 **DR. THOMAS STEEGER:**At this time, we're 

### 6 not proposing the use of doing genetic analyses to ensure that 

### 7 the bee populations that are typically used in these studies are 

### 8 drawn from healthy hives and presumably reflect a relatively 

### 9 broad genetic base. 

### 10 **DR. DANIEL SCHLENK:**Dr. Hunt. 

### 11 **DR. GREG HUNT:**I'm wondering, have there been 

### 12 any full field studies that have been classified as acceptable 

### 13 by the EPA? 

### 14 **DR. THOMAS STEEGER:**We have classified 

### 15 studies as acceptable as having met the standard or the 

### 16 guideline requirement for conducting a full field study, but as 

### 17 was pointed out by Joe DeCant, the 850.3040 guideline is 

### 18 relatively broad and does not specify specific expectations that 

### 19 would have to be met or specific criteria that would have to be 

### 20 met to downgrade a study unless things that have happened, such 

### 21 as contamination, have reached a point that the ability of the 

### 22 study to differentiate a treatment level effect has been 

### 23 completely compromised. 

### 24 If the study is scientifically unsound it will 

### 25 be downgraded, but we have accepted field studies; but primarily 

### 26 because there aren't strong criteria for potentially downgrading

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1 those, except best professional judgment that the scientific 

### 2 integrity has been compromised. 

### 3 **DR. GREG HUNT:**Thank you. That answers my 

### 4 question. I was just concerned about the level of cost and time 

### 5 in doing these studies. 

### 6 **DR. DANIEL SCHLENK:**Mr. Pistorius. 

### 7 **MR. JENS PISTORIUS:**Would it be possible to 

### 8 skip Tier II and go to Tier III directly? 

### 9 **DR. THOMAS STEEGER:**Again, the transition 

### 10 from screening level assessments to higher tier assessments are 

### 11 a risk management decision. It is entirely possible to 

### 12 transition from a screening level to a full field study; 

### 13 however, the intent, as you move up in refinement, is to refine 

### 14 the uncertainties; and I'm not sure that you would have been 

### 15 able to do that by transitioning quickly to a full field study 

### 16 from what could be gleaned from laboratory studies, unless you 

### 17 had available to you open literature studies or incident data 

### 18 that would enable you to do those types of refinements so that 

### 19 you can better fine tune the risk hypothesis that's being tested 

### 20 at the full field level. 

### 21 **MR. JENS PISTORIUS:**Another one. What would 

### 22 be the basis for election of which Tier II test has to be 

### 23 conducted, taking into account the different limitations of the 

### 24 described methods? 

### 25 When would you say use this test, use this 

### 26 test, free feeding, Oomen, et al. and all these different kind

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1 of -- 

### 2 **DR. THOMAS STEEGER:**I think that you're 

### 3 asking some very excellent questions and we're hoping that the 

### 4 SAP can provide insight on that. Again, the planning dialogue 

### 5 that takes place as part of the risk assessment process -- the 

### 6 risk assessor will inform the risk manager of what kind of 

### 7 uncertainties there are and we attempt to work with the 

### 8 regulated community in terms of how best to address those 

### 9 uncertainties. 

### 10 So whether you use a semi-field tunnel study 

### 11 or whether you use a feeding study or you use the Oomen method, 

### 12 those are all on the table for discussion in terms of what 

### 13 direction the Agency will actually require the registrant to 

### 14 take. 

### 15 **DR. DANIEL SCHLENK:**Okay. Dr. Fefferman. 

### 16 **DR. NINA FEFFERMAN:**Forgive me if this is a 

### 17 really silly question, it relies on two things I don't know of 

### 18 honey bee physiology and basic toxicology. 

### 19 The discussions I've been hearing about 

### 20 toxicity levels, again, I know you're mainly focusing on the 

### 21 acute side of things, but on the chronic side, the flip side of 

### 22 exposure, of course, is elimination. 

### 23 I was wondering whether or not these are 

### 24 things such that once they're ingested the stay in the bee 

### 25 permanently or if there is a rate of elimination kind of 

### 26 question that we should be asking. And if so, if there's any

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1 room in the kind of toxicity studies that you're running to look 

### 2 at the durational dose response curves and dose maintenance 

### 3 curves in a bee over its individual life and then possibly life 

### 4 of a colony. 

### 5 I'm thinking specifically of things that might 

### 6 be pulse treatment types of pesticides and whether or not that 

### 7 will actually be a constant daily level or whether or not as 

### 8 long as they survive the main pulse and we don't pulse them 

### 9 again until that's out of their system that might be okay. That 

### 10 kind of thing. 

### 11 **DR. THOMAS STEEGER:**I think you raised 

### 12 some very interesting questions. The screening level assessment 

### 13 is obviously testing or focused on acute toxicity and the 

### 14 exposure estimates that we're using are based on the presumption 

### 15 that none of the compound is degrading, so that's why they're 

### 16 very conservative assessment estimates. But as you move up in 

### 17 higher levels of refinement and the duration of these studies is 

### 18 more protracted, presumably you are able to incorporate how the 

### 19 chemical is moving through the colony, through the animal and 

### 20 looking at its potential to rebound from the effects that 

### 21 might've been observed at lower level studies. 

### 22 So actually attempting to isolate out the 

### 23 formation and decline curves of compounds within the bee can be 

### 24 a problem because you're going to have to be manipulating the 

### 25 colony quite a bit. And as Joe DeCant had indicated, the more 

### 26 that you manipulate the colony the more variability it's likely

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1 that you're going to introduce into it. 

### 2 So our focus at this time is trying to collect 

### 3 data points that provide us a sense for the colony level 

### 4 response, as opposed to the individual bee response and under as 

### 5 natural conditions as possible without over manipulating the 

### 6 colonies to try to get at those answers. 

### 7 **DR. DANIEL SCHLENK:**Do you have something 

### 8 to add? 

### 9 **MR. KEITH SAPPINGTON:**I just wanted to add 

### 10 that some of the characteristics of the compound -- while I 

### 11 agree in the Tier II and Tier III level you would have a 

### 12 long-term exposure -- but some of the characteristics of the 

### 13 compound, particularly its persistence in bioaccumulative 

### 14 abilities or characteristics would be sort of indicators, up 

### 15 front, that you might consider longer term accumulation within 

### 16 the organism. We held a separate SAP about four years ago on 

### 17 this whole process of how you do risk assessment for persistent 

### 18 bioaccumulative and toxic chemicals. 

### 19 With that said, we have a process for aquatic 

### 20 organisms that relies on bioaccumulation modeling as well as 

### 21 measured information in bioaccumulation tests. And there are 

### 22 some models that we use as a kind of a screen, up front, in 

### 23 problem formulation for terrestrial organisms. But to my 

### 24 knowledge, I have not seen a model yet that looks specifically 

### 25 at the toxicokinetics and accumulation within the bee; but at 

### 26 least we might get an indication early on.

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1 **DR. DANIEL SCHLENK:**I think Dr. Potter 

### 2 had a -- 

### 3 **DR. THOMAS STEEGER:**This is Tom Steeger. 

### 4 I'm sorry. 

### 5 **DR. DANIEL SCHLENK:**Go ahead. One more. 

### 6 **DR. THOMAS STEEGER:**I'd just like to add 

### 7 onto a question that had been asked earlier by Dr. Potter 

### 8 regarding the utility of the higher tier field studies. My 

### 9 interpretation of your question was relative to full hive 

### 10 studies. 

### 11 If your question was relative to exposure, 

### 12 higher tier exposure studies, those studies have proven to be 

### 13 very effective means of documenting the level of exposure that 

### 14 colonies or individual bees are exposed to. 

### 15 We've been working collaboratively with the 

### 16 California Department of Pesticide Regulation and with Health 

### 17 Canada Pest Management Regulatory Agency to collect those 

### 18 studies from field-based information and they are very useful 

### 19 for documenting exposure. 

### 20 **DR. DANIEL SCHLENK:**Dr. Potter. 

### 21 **DR. THOMAS POTTER:**I guess as a follow-up, 

### 22 are information from those studies imbedded in this risk 

### 23 assessment process that we're looking at here today? 

### 24 **DR. THOMAS STEEGER:**We are proposing that 

### 25 was depicted in the flow chart that Keith Sappington discussed 

### 26 and as Chris Garber discussed as well in her presentation that

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1 -- it's not up on the board -- but Box 6(a) is where you're 

### 2 evaluating potential exposure through RQ values, through 

### 3 field-based studies and those estimates of exposure that are 

### 4 actually measured values feed back into the calculation of RQ 

### 5 values that would hopefully, potentially reduce the RQ value 

### 6 below the level of concern. 

### 7 So what is inherent in this risk assessment 

### 8 framework that we are proposing is that field level measurement, 

### 9 whether they're collected either from the semi-field environment 

### 10 or from a full field study where you're actually focusing on 

### 11 exposure, you're measuring residues in pollen and nectar, would 

### 12 then be used at the numerator in the risk quotient that we'll be 

### 13 talking about in the next section that would be able to 

### 14 potentially -- at a screening level, where you're still 

### 15 quantifying risk -- to get risk quotients below LOCs and be able 

### 16 to come to a presumption of minimal risk. 

### 17 **DR. THOMAS POTTER:**Can I follow up on that? 

### 18 **DR. DANIEL SCHLENK:**Sure. 

### 19 **DR. THOMAS POTTER:**In looking through the 

### 20 White Paper, I was struck by what I perceived to be a lack of 

### 21 information or good quality data regarding concentrations of 

### 22 active ingredients in pollen and nectar so that to a large 

### 23 degree, we're dependent on a limited number of studies to 

### 24 compare the proposed exposure levels to. 

### 25 If there is additional information that's been 

### 26 submitted as part of the registration process, it certainly

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1 would seem to me to be very helpful, in terms of supporting this 

### 2 entire process to somehow get that data out to the light of day. 

### 3 **DR. THOMAS STEEGER:**I think that Dr. 

### 4 Berenbaum raised that same issue in terms of what appears to be 

### 5 relatively dated information. Those are published studies, but 

### 6 the Agency does have registrant-submitted studies that have been 

### 7 conducted either on the initiative of the regulated community or 

### 8 in response to data call-ins, such as that of California's, that 

### 9 are providing that very information and we deem it to be high 

### 10 quality information that would enable us to test the 

### 11 assumptions. 

### 12 In fact, we've compared some of these measured 

### 13 residues against the model estimates, the Chemical X or Chemical 

### 14 1, the undisclosed chemical. That information is still 

### 15 considered confidential business information, but those are 

### 16 actually measured values that some of these model estimates and 

### 17 default values are being compared to. 

### 18 **MS. KRISTINA GARBER:**Just to add a little bit 

### 19 more, we did actually look at all the registrant-submitted data 

### 20 that we had for pollen and nectar and we incorporated everything 

### 21 we could in the White Paper. 

### 22 **DR. THOMAS POTTER:**I will note that if you 

### 23 look at the executive summary of the Pellston conference -- I 

### 24 believe it's in the European Risk Assessment document as well -- 

### 25 that identified the need for data that can be used to support 

### 26 the risk assessment process as a critical need. So I'll leave

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1 it at there. We can go around in circles with that, but thank 

### 2 you. 

### 3 **DR. DANIEL SCHLENK:**Any other questions 

### 4 related to the presentation? 

### 5 (No response.) 

### 6 Okay. Dr. Steeger, do you want to move us 

### 7 ahead? 

### 8 **DR. THOMAS STEEGER:**Thank you for this 

### 9 opportunity to address the Scientific Advisory Panel. Again, my 

### 10 name is Tom Steeger. I'm a biologist and I'm a senior science 

### 11 advisor in the Environmental Fate and Effects Division. 

### 12 In this presentation, I will discuss the risk 

### 13 characterization component of the risk assessment process and 

### 14 how risk characterization integrates exposure and effect 

### 15 estimates with other lines of evidence to provide risk managers 

### 16 with an understanding of potential risk from the proposed use of 

### 17 the chemical. 

### 18 I'll also discuss how the risk 

### 19 characterization articulates the assumptions, limitations, and 

### 20 uncertainties associated with the available data. 

### 21 The risk characterization is intended to 

### 22 provide risk managers with an overall understanding of potential 

### 23 risks to the environment. In doing so, the assessment must be 

### 24 transparent, that is, it must be explicit with respect to the 

### 25 process that was followed. The risk assessment must be clear 

### 26 and easy to understand and consistent with Agency guidance and

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1 reasonable such that the risk manager and any stakeholder can 

### 2 understand how conclusions were arrived at, such that the risk 

### 3 assessment could be repeated and reach a relatively similar 

### 4 conclusion given the same process, underlying data and 

### 5 assumptions. 

### 6 As was discussed in the preceding 

### 7 presentations by Tom Moriarty and Keith Sappington this morning, 

### 8 the risk assessment process consists of three phases or steps: 

### 9 the problem formulation phase, the analysis phase, where effects 

### 10 are characterized and exposure is characterized, and that's 

### 11 followed by the risk characterization phase. 

### 12 Although the flow chart on the left is 

### 13 depicted as unidirectional, as Keith had pointed out earlier, 

### 14 the various phases in the risk assessment are intended to be 

### 15 iterative and as more information becomes available through the 

### 16 analysis phase, the problem formation may evolve. 

### 17 Communication with risk manager is an 

### 18 essential component of the risk assessment process since 

### 19 potential mitigation measures or other additional data can be 

### 20 evaluated in the context of how exposure estimates may be 

### 21 affected. 

### 22 Once the analysis phase is concluded, the risk 

### 23 assessment process proceeds to the risk characterization phase. 

### 24 This phase consists of two components: the risk estimation and 

### 25 the risk description. 

### 26 During risk estimation step of risk

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1 characterization, risk estimates are calculated. Risk estimates 

### 2 are based on a deterministic approach using point estimates or 

### 3 providing that there are sufficient data, estimates can be based 

### 4 on a distributional approach. 

### 5 Although not discussed extensively in the 

### 6 White Paper, distribution-based approaches, as opposed to point 

### 7 estimate-based approaches, can be use to characterize both 

### 8 exposure and effects and joint probability curves constructed to 

### 9 then estimate the likelihood and magnitude of an adverse effect. 

### 10 However, in today's presentation, we are 

### 11 focused on point estimate-based approach, which compares risk 

### 12 estimates to levels of concern and the potential for risk as 

### 13 articulated in terms of whether the level of concern is exceeded 

### 14 or not. 

### 15 The proposed framework for estimating risk at 

### 16 a screening level is consistent with the process currently used 

### 17 by OPP to estimate risk to other taxa, and is based on the 

### 18 concept of risk being a function of both exposure and toxicity. 

### 19 Point estimate of environmental concentrations, such as those 

### 20 described earlier by Kris Garber, are compared to point 

### 21 estimates of toxicity endpoints discussed by Joe DeCant. 

### 22 The quotient of the ratio of exposure to 

### 23 effects is the risk quotient. As discussed by Kris Garber, at 

### 24 the screening level, point estimates of exposure are initially 

### 25 based on relatively conservative models or default values. 

### 26 However the risk assessment process is

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1 intended to be iterative, and as discussed by Reuben Baris, 

### 2 exposure values used to generate RQs can be further refined 

### 3 through the use of measured values obtained through other 

### 4 studies, typically submitted to EPA, such as magnitude of 

### 5 residue studies or through targeted monitoring studies of 

### 6 residues and foliage or in pollen and nectar. 

### 7 Table 22 of the White Paper provides a summary 

### 8 of the proposed exposure and effect point estimates used to 

### 9 calculate risk quotients and the screening level risk assessment 

### 10 for pesticides applied folairly, as seed treatments, soil 

### 11 treatments, and tree trunk injections. 

### 12 This is an excerpt from the table and it's 

### 13 taken from Table 22, for pesticides applied to seed treatments, 

### 14 the dietary-based RQ values for adults is based on the products 

### 15 of the Briggs model estimated environmental concentration, 

### 16 multiplied by .29 grams per day, serving as a numerator, divided 

### 17 by the acute oral LD50 value for individual adult bees. 

### 18 RQ values estimated in the risk estimation 

### 19 phase of the risk characterization are compared to the Agency's 

### 20 level of concern. Levels of concern represent the Agency's 

### 21 interpretive policy and are used to analyze potential risk to 

### 22 non-target organisms and the need to consider regulatory action. 

### 23 24 These criteria are used to indicate when a 

### 25 pesticide is used as directed on the label, has the potential to 

### 26 cause adverse effects on the non-target organisms.

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1 Historically, the Environmental Fate and Effects Division has 

### 2 used the default probit dose response slope of 4.5 to estimate 

### 3 acute effects to organisms; however, the level of concern for 

### 4 estimating acute risk to honey bees is based on an analysis of 

### 5 honey bee acute contact and acute oral toxicity studies that had 

### 6 been submitted to EPA. 

### 7 This analysis resulted in a mean probit dose 

### 8 response slope for acute contact toxicity at 3.93 and a mean 

### 9 probit dose response slope for acute oral toxicity studies of 

### 10 3.4. The Environmental Fate and Effects Division has relied on 

### 11 the Individual Effect Chance model, which is abbreviated the 

### 12 IEC, to estimate the likelihood of an effect, for example, acute 

### 13 mortality. 

### 14 This model uses the calculated risk quotient 

### 15 and the probit dose response slope that integrates those values 

### 16 to derive the probability of an individual mortality. However, 

### 17 the model can be used in reverse to estimate the risk quotient 

### 18 value that would be needed to result in a particular magnitude 

### 19 of an effect, given a specific dose response slope. 

### 20 Since data are available for both acute oral 

### 21 and acute contact lethality studies, both sets of data were 

### 22 combined. Rather than using the mean dose response values, 

### 23 though, the median dose response slope across all acute contact 

### 24 and oral toxicity studies was used as a measure of central 

### 25 tendency and this median dose response slope is 3.2. 

### 26 Using the individual effect calculated model,

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1 the level of concern was then back calculated, which was a 

### 2 result in 10 percent mortality based on the median dose response 

### 3 slope of 3.2. This slide depicts Table 24 from the White Paper, 

### 4 summarizing that the median slope of 3.2 and the likelihood of 

### 5 10 percent mortality that is in effect, the LOC of 0.4 is 

### 6 necessary. 

### 7 The 10 percent level of effect, that is the 

### 8 acute mortality, is based on the maximum level of mortality 

### 9 permitted in controls. That is, untreated studies of acute oral 

### 10 and contact toxicity based on current guideline recommendations. 

### 11 Although the chronic toxicity study guidelines 

### 12 are not currently available for honey bees the proposed chronic 

### 13 risk level of concern is 1) since the RQ value for estimating 

### 14 chronic risk is the ratio of the maximum estimated exposure 

### 15 concentration to the no-observed adverse effect concentration; 

### 16 that is, the NOAEC, an LOC of one is consistent with that used 

### 17 for evaluating chronic risk to other taxa. 

### 18 As discussed by Mr. Sappington and as depicted 

### 19 in Figures 2 and 3 of the White Paper showing the proposed risk 

### 20 assessment framework for honey bees, the process is intended to 

### 21 be iterative. The slide depicts the risk estimation portion of 

### 22 the framework where risk quotients are calculated in Boxes 4(a) 

### 23 and 4(b), based on conservative exposure estimates derived in a 

### 24 screening level assessment. 

### 25 Charge Question 13 asks the SAP to provide 

### 26 comment on the exposure and effect data used to estimate risk

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1 and the acute and chronic risk LOCs that are in turn used to 

### 2 evaluate the risk quotients. Question 13 also asks the SAP to 

### 3 comment on how other line of evidence are integrated into the 

### 4 risk assessment during the second phase of risk 

### 5 characterization, that is, the risk description phase, which I 

### 6 will now discuss. 

### 7 The second component of risk characterization 

### 8 is risk description where RQ values are calculated in the risk 

### 9 estimation phases are considered in conjunction with other lines 

### 10 of evidence which may either support or refute these estimates, 

### 11 based on assessment endpoints. In this part of the risk 

### 12 characterization, the relevance of other lines of evidence are 

### 13 considered. 

### 14 The risk description builds on the RQ values 

### 15 calculated during the risk estimation phase of the risk 

### 16 characterization process. The risk description includes a 

### 17 discussion of possible refinements and exposure values -- 

### 18 discussed by both Chris Garber and Reuben Baris -- that lead to 

### 19 refined RQ values, using toxicity estimates for individual and 

### 20 adult bees. 

### 21 The quantitative information is then 

### 22 considered in the context of available studies that were 

### 23 discussed by Joe DeCant of the whole colony that may have been 

### 24 conducted under relatively controlled conditions, that is, 

### 25 semi-field studies, or under conditions that are intended to be 

### 26 more reflective of actual use conditions, that is, the full

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1 field Tier III studies. 

### 2 While adverse effects may be measured on 

### 3 individual bees and these effects quantified through risk 

### 4 quotients, the higher tier refinements are intended to answer 

### 5 whether adverse effects are observed at the whole colony level. 

### 6 The intent is for the risk assessor to determine whether or not 

### 7 there is a concordance of information to indicate whether there 

### 8 is a potential for adverse effects that could limit the extent 

### 9 to which the protection goals may be met. 

### 10 Thus, while RQ values may indicate that 

### 11 individual bees may be at risk from acute exposures to a 

### 12 pesticide, semi-field studies may indicate that no adverse 

### 13 effects occur at the whole colony level when the pesticide is 

### 14 applied to pollinator attractive crop at the maximum proposed 

### 15 application rate. 

### 16 Conversely, risk concerns, based on individual 

### 17 bees, may be supported by semi-field or full field studies, 

### 18 showing colony level effects. Other scientifically relevant 

### 19 information would be considered as well. 

### 20 The risk description integrates multiple lines 

### 21 of evidence in the context of assessment endpoints discussed by 

### 22 Tom Moriarty in the opening presentation. As such, the risk 

### 23 description is not intended to be simply a comparison of risk 

### 24 quotients to the levels of concern, rather, the quantitative 

### 25 estimates of risk, that is, the RQ values are integrated with 

### 26 other lines of evidence that may be available through higher

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1 tier guideline toxicity testing, non-guideline studies reported 

### 2 in open literature, and incident data. 

### 3 Other lines of evidence, such as incident 

### 4 data, available through the Office of Pesticides Program 

### 5 Incident Data System, the IDS, and the Ecological Incident 

### 6 Information System, the EIIS, provide a means for understanding 

### 7 the effects of chemicals under actual use conditions. 

### 8 As these additional lines of evidence are 

### 9 considered, the risk hypothesis and the conceptual model are 

### 10 refined. As discussed in the preceding presentations, the 

### 11 intent of the tiered risk assessment process is to provide 

### 12 increasingly realistic understanding of potential risk that 

### 13 reflects how the chemical will actually be used. 

### 14 For new chemicals, available data may be 

### 15 confined to guideline laboratory and field testing; however, for 

### 16 older chemicals, a wider array of data may be available and are 

### 17 considered, provided those data meet the standard established by 

### 18 EPA for inclusion in risk assessments. EPA has developed 

### 19 guidance for evaluating multiple lines of evidence, and an 

### 20 example of such guidance is discussed in the White Paper where 

### 21 elements of the endocrine disrupting screening program, weight 

### 22 of evidence document are articulated. 

### 23 Also, the Agency's risk characterization 

### 24 handbook developed by the EPA Office of Research and 

### 25 Development, also provides guidance on the factors that should 

### 26 be considered in risk characterization and which are discussed

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1 in the following slides. 

### 2 In considering the additional lines of 

### 3 evidence that may be available, the evaluation considers 1) the 

### 4 adequacy and quality of the data; 2) the degree and type of 

### 5 variability and uncertainty; and 3) the relationship of the 

### 6 evidence to risk assessment endpoints. 

### 7 With respect to the adequacy and the quality 

### 8 of the data, risk assessors consider whether the data quality 

### 9 objectives were followed. OPP has published formal guidance on 

### 10 the conduct and evaluation of guideline toxicity studies and has 

### 11 developed guidance for staff in evaluating the utility of open 

### 12 literature studies that may not adhere to testing guidelines. 

### 13 Just as the White Paper reflects and is 

### 14 consistent with the Agency's commitment to transparency, each 

### 15 risk description articulates what assumptions are made, where 

### 16 there are data gaps and what type of limitations may be present 

### 17 when non-guideline or supplemental data are used. As such, the 

### 18 experimental designs have been considered when evaluating the 

### 19 utility of the data for characterizing risk. 

### 20 As was discussed in the presentation by 

### 21 Christina Wendel, the Agency is clear as to what is considered 

### 22 in the risk assessment process and what is not considered and 

### 23 why. Also, when evaluating lines of evidence, degree and type 

### 24 of uncertainty have to be articulated. 

### 25 With respect to characterizing the degree and 

### 26 type of uncertainty and variability, variability refers to the

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1 differences in characteristics of individuals or exposure, 

### 2 reflecting inherent variety in test organisms or exposure 

### 3 scenarios that can be quantified. 

### 4 With respect to characterizing uncertainty, 

### 5 the risk characterization should include a discussion of what is 

### 6 not known, that is, of the uncertainties that may be associated 

### 7 with the available data. Also in evaluating lines of evidence, 

### 8 the relationship of the evidence to risk assessment has to be 

### 9 articulated. 

### 10 Relationship of available evidence to risk 

### 11 assessment can draw from previous assessments on the pesticide 

### 12 or similarly structured pesticides and/or risk assessments 

### 13 conducted by other organizations, such as Pest Management 

### 14 Regulatory Authority -- or Agency, the Australian APVMA, and the 

### 15 European Food Safety Authority. Plus, the risk characterization 

### 16 must reflect current agency policy regarding the use and 

### 17 interpretation of data. 

### 18 As discussed earlier by Tom Moriarty, the risk 

### 19 assessment framework discussed in the White Paper is intended to 

### 20 apply to honey bees; however, given that laboratory studies are 

### 21 based on individual bees and that higher tier studies, discussed 

### 22 by Joe DeCant examine both individual bees and colony level 

### 23 effects, the assessment may provide insight on potential 

### 24 exposure to and effects of the pesticide to solitary and social 

### 25 non-Apis bees as well. 

### 26 In addition to measurement endpoints that are

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1 directly related to assessment endpoints of a impaired growth, 

### 2 survival and reproduction, the risk description would also 

### 3 provide a discussion of sublethal effects that may have been 

### 4 measured in the various studies that have been evaluated, and 

### 5 the uncertainties associated with how these measurement 

### 6 endpoints may be related to the assessment endpoints. 

### 7 Finally, the risk description should, to the 

### 8 extent possible, discuss the potential adversity of risks in 

### 9 terms of the nature and intensity of the effects, where there is 

### 10 evidence to suggest temporal or spacial trends, and whether the 

### 11 data indicate that recovery may be possible. 

### 12 In the following presentation by Reuben Baris, 

### 13 the relative conservatism of the proposed risk assessment 

### 14 process will be briefly discussed. Also, as multiple lines of 

### 15 evidence are intended to be covered in the risk description, the 

### 16 white paper discusses the potential utility of colony level 

### 17 models to estimate potential effects of pesticides. Therefore, 

### 18 Keith Sappington will provide a presentation on how these models 

### 19 can inform the risk assessment process. Any questions? 

### 20 **DR. DANIEL SCHLENK:**Wow. Thanks, Dr. 

### 21 Steeger. Any questions for Dr. Steeger? Dr. Pistorius? 

### 22 **MR. JENS PISTORIUS:**I have one. Has there 

### 23 been an exercise done with a new proposed risk assessment 

### 24 process? How many substances would fail or pass compared to the 

### 25 old risk assessment? Is it many more or few more or all? 

### 26 **DR. THOMAS STEEGER:**This is Tom Steeger.

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1 That is a most excellent question and you're going to get an 

### 2 answer to it in the next presentation. 

### 3 **DR. DANIEL SCHLENK:**Any other questions? 

### 4 Okay. We're running a bit behind. I want to take a poll of the 

### 5 panel here. Who would rather take about a five minute break or 

### 6 keep going? We have two more presentations left to go. So, in 

### 7 favor of keep going, hands; and five minute break? Okay. Looks 

### 8 like we're going to keep going. Okay. With that, Reuben Baris, 

### 9 I guess you're up next. 

### 10 **MR. REUBEN BARIS:**Thank you very much. My 

### 11 name is Reuben Baris. The information presented in this 

### 12 presentation was not included in the White Paper, but is 

### 13 intended to provide contextual information about the efficiency 

### 14 of the Tier I screen as presented in earlier presentations. As 

### 15 presented earlier, the purpose of the Tier I screen is to 

### 16 efficiently identify those pesticides that do not pose a risk to 

### 17 honey bees, so that researches can be focused on pesticides that 

### 18 potentially have risks. 

### 19 Available acute oral and contact toxicity for 

### 20 adult worker bees from unpublished registrant studies were 

### 21 compiled for this analysis. This is a partial examination and 

### 22 is based only on available toxicity data for adult worker bees. 

### 23 24 Individual chemical risk quotients were 

### 25 calculated and compared to the proposed level of concern that 

### 26 was presented just now by Dr. Steeger. This effort was

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1 completed to provide some context on the efficiency of the 

### 2 initial exposure screen using the most conservative assumptions 

### 3 of exposure. The Tier I screen for foliar applications were run 

### 4 for each chemical using the maximum foliar application permitted 

### 5 on the label. The Briggs model was not included in this 

### 6 analysis because there is limited oral toxicity information for 

### 7 adult worker bees for chemicals where soil applications are 

### 8 permitted on the label. Therefore, this analysis applies to 

### 9 foliar applications only. 

### 10 A subset of adult worker acute contact 

### 11 toxicity data that cover a breadth of chemical categories, 

### 12 chemical groups and modes of action were used in this analysis. 

### 13 Shown here, the number of chemicals in each category is similar, 

### 14 and a few examples of the types of chemical groups are shown in 

### 15 the parentheses. 

### 16 An attempt was made within each category to 

### 17 use chemicals from different groups and modes of action in order 

### 18 to remove potential bias towards a specific chemical activity. 

### 19 All available data from unpublished registrants submitted acute 

### 20 oral toxicity studies for adult worker bees were used in the 

### 21 acute oral analysis. Many of the chemical groups and modes of 

### 22 action represented in the acute contact analysis were also 

### 23 represented in this acute oral analysis. 

### 24 This table and the following slide represent a 

### 25 snapshot of the Tier I screen based on available acute contact 

### 26 toxicity data. As was presented by Kris Garber this afternoon,

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1 the Koch and Weisser approach with honey bee specific data is 

### 2 the agency's proposed approach for the Tier I contact exposure 

### 3 screen. 

### 4 Shown here, fewer insecticides make it through 

### 5 the screen and would likely require additional refinement. 

### 6 However, herbicides and fungicides generally pass the screen 

### 7 with approximately 32 and 4 percent respectively requiring 

### 8 additional evaluation or refinement. 

### 9 Fewer insecticides pass the initial Tier I 

### 10 screen compared to herbicides and fungicides. This is expected 

### 11 because insecticides are more likely to directly affect insects 

### 12 such as honey bees. It is clear from the table shown there that 

### 13 while greater than 85 percent of the insecticides evaluated did 

### 14 not pass the screen, the evaluation was based on acute toxicity 

### 15 studies with young adult bees. 

### 16 Larval toxicity testing may result in RQ 

### 17 values that exceed the level of concern of 0.4. However, even 

### 18 for some insecticides, it is possible that bees are not 

### 19 particularly sensitive, and these chemical will likely have RQ 

### 20 values that are below the LOC, just as not all plants are 

### 21 sensitive to herbicides. 

### 22 Since fungicides and herbicides tend to target 

### 23 fungi or plants, it is less likely that they will directly 

### 24 affect insects. This analysis was also completed with the T-REX 

### 25 contact approach, and similar results were observed. 

### 26 Again, as was expected in this acute oral

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1 analysis for adult worker bees and shown previously on the 

### 2 previous slide by the contact exposure analysis, fewer 

### 3 insecticides pass the initial Tier I screen compared to 

### 4 herbicides and fungicides. There are a limited number of 

### 5 fungicides that are included in this analysis because of limited 

### 6 oral toxicity data that are available for adult worker bees. 

### 7 But again, as shown by these preliminary results, the Tier I 

### 8 screen is functioning well as a screen by identifying the 

### 9 chemicals that need additional refinement and screening out the 

### 10 majority of non-insecticides that are not likely to be a risk 

### 11 concern. 

### 12 This analysis was an exercise to illustrate 

### 13 the decision point of Box 5(a) and (b) of the proposed decision 

### 14 tree. That is, does the contact or oral RQ exceed the level of 

### 15 concern? It provides context for the efficiency of the Tier I 

### 16 screen. The analysis included a variety of chemical groups and 

### 17 modes of action for contact and oral toxicity, identifying those 

### 18 chemicals that require additional refinements. These 

### 19 refinements, Box 6 of the decision tree would include working 

### 20 with risk managers to refine the Tier I exposure assessment. 

### 21 Therefore, the conclusion is that the Tier I 

### 22 approach is functioning as expected as an initial screen. It is 

### 23 important to keep in mind that these preliminary results are 

### 24 intended to give a very rough example that the screen in not 

### 25 serving as a wall for all classes of pesticides, but rather, it 

### 26 is identifying those chemicals that are most likely to need

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1 additional review. In doing so, only model-estimated exposure 

### 2 values have been evaluated. 

### 3 It is possible that measured exposure values 

### 4 could result in a reduction in risk estimates such that further 

### 5 refinements -- that is higher tiered testing -- may not be 

### 6 required. However, as mentioned by Dr. Steeger in the previous 

### 7 presentation, multiple lines of evidence are used in determining 

### 8 the risk to honey bees. 

### 9 The next presentation given by Mr. Sappington 

### 10 will discuss colony level models and how they can be used in the 

### 11 risk characterization process. Thank you for your time. 

### 12 **DR. DANIEL SCHLENK:**Questions? Dr. 

### 13 Berenbaum? 

### 14 **DR. MAY BERENBAUM:**While fungicides might 

### 15 themselves not be considered to present as greater risk, there 

### 16 is increasing evidence that in combination fungicides can 

### 17 exacerbate risks. Were any studies conducted on combinations or 

### 18 synergistic interactions? 

### 19 **DR. THOMAS STEEGER:**This is Tom Steeger. 

### 20 That as Reuben pointed out towards the end of his presentation, 

### 21 the risk quotient isn't viewed in isolation and other lines of 

### 22 evidence would be considered. And you're absolutely right, 

### 23 there is open literature pointing to potential interactive 

### 24 effects and we have required studies to address those 

### 25 uncertainties, even for chemicals that had initially RQ values 

### 26 that would have been below the LOC.

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1 **DR. DANIEL SCHLENK:**Okay. Any other 

### 2 questions? Yeah, Dr. Hunt? 

### 3 **DR. GREG HUNT:**Well, along those same lines, 

### 4 some formulations come as a package, such as seed treatments. 

### 5 Is there any consideration that that might be required of 

### 6 registrants to - I know you can't test every possible 

### 7 interaction, but there are some common formulations that could 

### 8 be tested. 

### 9 **DR. THOMAS STEEGER:**There is a terminology 

### 10 difference that we deal with. When we talk about formulations, 

### 11 oftentimes what the agency is meaning is chemicals that are 

### 12 co-formulated into a single product as opposed to what you might 

### 13 see happen out at the field level where are tank mixes, and 

### 14 these aren't really formulations. These are brews, for lack of 

### 15 a better term, that are used. 

### 16 Where we have an indication that a particular 

### 17 formulation could potentially be more toxic to a taxa, whether 

### 18 it be honey bee or any taxa that we're evaluating, we may call 

### 19 in - again it's a risk management call - data to address that 

### 20 uncertainty. 

### 21 **DR. DANIEL SCHLENK:**Okay. No further 

### 22 questions? Our last presentation of the day, no pressure there 

### 23 Keith. 

### 24 **MR. KEITH SAPPINGTON:**Thanks for bringing up 

### 25 brew. This will make me go faster. 

### 26 Good afternoon. Again, my name is Keith

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1 Sappington with the Environmental Fate and Effects Division. I 

### 2 will be changing gears a little bit here in the context of our 

### 3 previous discussions and talking about the potential use of 

### 4 mathematical models in the context of pesticide risk assessment 

### 5 for honey bees. 

### 6 For convenience, we've termed these as colony 

### 7 level models, which mean that the models focus on describing the 

### 8 dynamics of a single colony, not a population of multiple 

### 9 colonies. This information is found in section 5.4 of the White 

### 10 Paper. I would also like to acknowledge the work of Kris Garber 

### 11 who helped greatly in the review, providing additional support 

### 12 for this topic in the White Paper. 

### 13 So, first, I will briefly describe some of the 

### 14 reasons why we may be interested in colony-level models in the 

### 15 context of pesticide risk assessment with honey bees and then I 

### 16 will discuss their current state of the science. Finally, I 

### 17 will conclude with a brief description of selected models that 

### 18 we identified in the literature. 

### 19 There are a number of reasons why conceptually 

### 20 we would be interested in these models, and I'm sure they are 

### 21 obvious to the panel at this point. First of all, these models 

### 22 enable the dynamics of a honey bee population to be measured. 

### 23 These dynamics is shaped numerous interdependent processes, 

### 24 which can be partially informed and managed through biological 

### 25 feedback loops. 

### 26 For example, excessive loss of foragers may be

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1 partially compensated for by an earlier recruitment of hive bees 

### 2 into the forager workforce. Such a change may compensate for 

### 3 the loss of foragers in the short term, but may in the longer 

### 4 term lead to adverse effects on the colony through a reduction 

### 5 in brood rearing efficiency as well as a reduction in worker 

### 6 longevity. 

### 7 While empirical studies may capture the net 

### 8 effect of these interdependent processes on overall colony 

### 9 strength, a properly constructed colony-level model could 

### 10 provide a means for understanding these relationships, and 

### 11 therefore enable one to extrapolate observed effects over 

### 12 perhaps longer period of time and possibly under different 

### 13 environmental conditions. 

### 14 Second, colony level models could help in 

### 15 interpreting sublethal effects, which we have discussed on 

### 16 repeated occasions in earlier presentations. And in particular, 

### 17 for linking sublethal effects quantitatively on individual bees 

### 18 to effects at the whole colony level. 

### 19 Third, colony level models could also help in 

### 20 a design of empirical studies such as the Tier II and Tier III 

### 21 field studies described by Joe DeCant. For example, they could 

### 22 be used in addressing study design elements such as when to 

### 23 conduct a study, at what spacial scale, and even help with 

### 24 deciding on the degree of replication necessary. They also can 

### 25 help in identifying the duration that may be needed for a 

### 26 particular semi-field or field study.

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1 Fourth, such models could also assist in 

### 2 interpreting the results from the Tier II and Tier III studies, 

### 3 such as accounting for the seasonal effects on measurement 

### 4 endpoints or other factors like the impact of the weather on 

### 5 model results, or on observed results. 

### 6 In the context of the proposed risk assessment 

### 7 scheme, colony-level models could enable one to interpret 

### 8 results of toxicity studies and integrate these results from 

### 9 different risk assessment tiers, all the way from Tier I at the 

### 10 individual level through Tier II and Tier III. 

### 11 Finally, such models could potentially 

### 12 incorporate not only the effects of a single stressor, but those 

### 13 of multiple stressors, which may be either chemical or 

### 14 nonchemical in nature. 

### 15 The overall state of the science of 

### 16 colony-level models for honey bees was recently reviewed and 

### 17 published as part of an executive summary of the SETAC Pellston 

### 18 workshop. We've included the conclusions of this review here 

### 19 because it was conducted specifically in the context of their 

### 20 potential application in pesticide regulatory risk assessment. 

### 21 In this review, it was concluded that none of 

### 22 the available honey bee models seemed suitable for regulatory 

### 23 risk assessment for several reasons listed here. Collectively, 

### 24 these include a lack of explicit linkage of foraging bees to 

### 25 their surrounding landscape, insufficient testing of model 

### 26 results against empirical data, lack of a comprehensive

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1 sensitivity analysis in order to understand the impact of model 

### 2 parameters and assumptions, exclusion of multiple stressor 

### 3 impacts on model results and insufficiency documentation of 

### 4 model assumptions and/or algorithms in some cases. 

### 5 At this time, only the SETAC executive summary 

### 6 is available from the Pellston workshop. So, the detailed 

### 7 review specific to each model was not available to us. 

### 8 Because the details were not available at the 

### 9 time we were drafting the White Paper, we decided to conduct a 

### 10 very brief review of only selected models that we identified in 

### 11 the literature. As we point out in the White Paper, this review 

### 12 is not intended to be comprehensive or exhaustive. Instead, it 

### 13 reflects a sampling of more recent models that had been 

### 14 published for honey bees as a way for us to explore their 

### 15 potential utility in pesticide risk assessment. 

### 16 In this and the subsequent table, we summarize 

### 17 some of those attributes of the models we reviewed. The models 

### 18 are listed here from top to bottom, alphabetically by the lead 

### 19 author of the publication in which they appeared, and various 

### 20 attributes are listed at the top of this table. We note that 

### 21 inclusion of a model in this list does not constitute our 

### 22 endorsement nor does exclusion imply lack of endorsement. 

### 23 So, in looking at the first two columns, one 

### 24 can see that all but one of the models we considered address 

### 25 effects at the colony level with output including overall colony 

### 26 strength in composition. The model published by Makela et al.

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1 in 1993 is a true population model for honey bee in that it 

### 2 considers multiple honey bee colonies since it was designed to 

### 3 predict the impact and distribution of Africanized bees on the 

### 4 European honey bee. 

### 5 In the third column of this table, I've 

### 6 included some key model input parameters and model processes. 

### 7 I've also provided some of the unique features of each model in 

### 8 bold. For example, the model by Becher et al. in 2010 shown in 

### 9 the first row focuses on the distribution of comb temperature 

### 10 and how this in turn affects queen egg laying rate, brood 

### 11 development and overall colony strength. These authors 

### 12 acknowledge that the their model is, in effect, a partial model 

### 13 and it's not intended to address overall colony dynamics. 

### 14 Among other features, the BeePoP model 

### 15 published by DeGrandi-Hoffman shown in the second row includes 

### 16 an explicit representation of weather patterns in addition to 

### 17 modeling queen fertility as a function of queen age. 

### 18 The model by Khory et al. shown here in the 

### 19 third row was specifically designed to evaluate age-related 

### 20 division of labor among hive bees that is related to the onset 

### 21 of foraging. This model was designed to investigate 

### 22 specifically how the early onset of foraging could impact the 

### 23 survival of foragers as well as brood rearing efficiency. 

### 24 The inter-colony model, I just mentioned by 

### 25 Makela et al. in the fourth row contains some unique attributes 

### 26 such as bee swarming, inter-colony migration of bees and even

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1 changes in genotype. 

### 2 Perhaps the most comprehensive colony model in 

### 3 this collection in this table is the HoPoMo model published by 

### 4 Schmickl and Crailsheim in 2007. This model listed in the fifth 

### 5 row includes, among many other processes, foraging activity, 

### 6 food availability and even larval cannibalism and their impact 

### 7 on colony strength and composition. 

### 8 Lastly, the model in the last row by Thompson 

### 9 published in 2005 focuses on pesticide impacts on honey bee 

### 10 processes, including the queen egg laying rate, the onset of 

### 11 foraging activity, brood rearing and worker longevity. 

### 12 Other features of these models that we 

### 13 summarize include whether or not it is able to accommodate 

### 14 spatially explicit information, whether a formal sensitivity 

### 15 analysis was performed, the extent to which it has been verified 

### 16 using empirical data and its application to pesticide risk 

### 17 assessment. 

### 18 Several models we looked at were able to 

### 19 include some spatially explicit or geographic based information, 

### 20 although the BeePoP and HoPoMo models, this inclusion is limited 

### 21 to climate data, which can be made region or site-specific. 

### 22 Most models did not include a formal sensitivity analysis except 

### 23 for the BeePoP model, although some limited analysis of selected 

### 24 model input parameters was conducted in some other studies. 

### 25 In terms of verifying model projections with 

### 26 empirical data, only some of the models included a comparison of

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1 model results or model components to observe data. For example, 

### 2 Becher et al. compared their predicted timing of peak hive 

### 3 strength and minimum number of bees necessary for successful 

### 4 overwintering of hives to observe values. 

### 5 Khoury compared a predicted onset of foraging 

### 6 and the forager age to literature values. For the HoPoMo model, 

### 7 Schmickl and Crailsheim compared several model subcomponents as 

### 8 well as the entire model output. That is a number of bees to 

### 9 observed data. 

### 10 Lastly, only two of the models that we 

### 11 identified in this list included an explicit application to 

### 12 pesticide risk assessment. 

### 13 In this slide, we thought it would be useful 

### 14 to articulate some of the main attributes that we believe are 

### 15 desirable in order to apply a colony-level model in pesticide 

### 16 risk assessment. As indicated in the first bullet, it's is 

### 17 clearly important to account for the major processes that govern 

### 18 the dynamics of honey bee colonies, initially at the colony 

### 19 level and eventually at the population of inter-colony level. 

### 20 Second, the model should quantitatively link 

### 21 the various measurement endpoints from pesticide toxicity 

### 22 studies to the proposed assessment endpoints. For example, 

### 23 colony strength and survival, production of hive products. 

### 24 As indicated in the third bullet, such models 

### 25 should be able to be readily parameterized using existing 

### 26 biological and pesticide-specific exposure and effects data.

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1 Another desirable feature shown in the fourth 

### 2 bullet is the ability to tailor model results to a specific 

### 3 region of the country in order to account for differences in 

### 4 pesticide use patterns, crops, climate and even landscape 

### 5 features. 

### 6 Lastly, but no less important, the model must 

### 7 be scientifically defensible, highly transparent, well 

### 8 documented and publically available in keeping with the Agency's 

### 9 efforts to promote transparency with clear, concise and 

### 10 reasonable risk characterizations and tools used therein. 

### 11 We note in the footnote that EPA has published 

### 12 guidelines for the development of environmental models in 

### 13 general that are intended for use in regulatory activities. In 

### 14 addition, the European commission is developing several model 

### 15 guidelines specifically for mechanistically-based ecological 

### 16 effects models. 

### 17 Among other factors, a colony-level model 

### 18 should include a formal sensitivity analysis and uncertainty 

### 19 analysis in addition to verification of the model results to 

### 20 observe empirical data. 

### 21 So to conclude, the SETAC Pellston workshop 

### 22 participants indicate the currently colony level simulation 

### 23 models are not ready for regulatory application. This 

### 24 conclusion seems consistent with our cursory review of a limited 

### 25 subset of models. However, once these models are fully 

### 26 developed and evaluated, we think the colony-level models could

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1 provide an important line of evidence in the risk 

### 2 characterization of pesticide effects on honey bees, and 

### 3 potentially help to identify adverse outcome pathways across 

### 4 multiple levels of biological organization. 

### 5 As we pointed out, they could be extremely 

### 6 valuable in the problem formulation phase in an assessment in 

### 7 the design and interpretation of field studies and in 

### 8 integrating pesticide effects and exposure data generated at 

### 9 multiple risk assessment tiers and at various levels of 

### 10 biological organization. 

### 11 Therefore as indicated in the final charge 

### 12 question number 14, we are interested in SAP feedback on the 

### 13 concept of using colony-level models in pesticide ecological 

### 14 risk assessment. In other words, is this concept worth pursuing 

### 15 by the pesticide regulatory agencies represented? 

### 16 In addition, we would like SAP feedback on the 

### 17 current state of the science of honey bee models in relation to 

### 18 their potential use and application in pesticide risk 

### 19 assessment. 

### 20 And finally, we would appreciate your input on 

### 21 the most important model design elements that we should consider 

### 22 as part of any review of a colony level model for application in 

### 23 pesticide risk assessment. Thank you very much. 

### 24 **DR. DANIEL SCHLENK:**Any questions? Dr. 

### 25 Fefferman? 

### 26 **DR. NINA FEFFERMAN:**Just as a very high-level

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1 cursory question in shifting from the White Paper discussion to 

### 2 your presentation today, there was a shift from a discussion of 

### 3 general models for colony-level use to simulation models. I was 

### 4 wondering if that was purposeful and if you were interested in 

### 5 analytic mathematical models and formulations as well as 

### 6 simulation-based methods? 

### 7 **MR. KEITH SAPPINGTON:**Could I ask you to 

### 8 clarify general models as compared to mathematical models? 

### 9 **DR. NINA FEFFERMAN:**Sorry, so in my mind sort 

### 10 of mathematical analytic models live on one side and simulation 

### 11 models live on another. So, simulation models are ones where 

### 12 there are algorithms for action, and from those you get 

### 13 numerical solutions, whereas mathematical models are ones where 

### 14 by understanding the interactions of systems of equations, you 

### 15 do things like generate threshold behavior or equilibrium 

### 16 states. Things that don't involve, for example, running Monte 

### 17 Carlo analyses in the same way as outcome measurements. 

### 18 So, simulation models produce data. Analytic 

### 19 models produce sort of dynamic insight, and I was wondering if 

### 20 you are specifically restricting yourself to simply the 

### 21 simulation methods or if you're also interested in methods that 

### 22 provide close form equation type understanding. 

### 23 **MR. KEITH SAPPINGTON:**Keith Sappington. 

### 24 There was not an intentional restriction to the simulation 

### 25 models, although they were attractive because of the analytical 

### 26 components and the type of measuring endpoints included and the

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1 outcome aligning very closely to our assessment endpoints. With 

### 2 that said, if the analytical versions of the models could 

### 3 provide us insights, for example, the thresholds that need to be 

### 4 maintained for long-term hive success, we would welcome input on 

### 5 that. 

### 6 **DR. NINA FEFFERMAN:**Thank you. I'm sure I'll 

### 7 talk about this more with you when we get to the specific 

### 8 modeling section of the discussion, but thanks. That's great. 

### 9 **DR. DANIEL SCHLENK:**Dr. Schwab? 

### 10 **DR. PAUL SCHWAB:**Given the fact that the 

### 11 number of times we have talked about chronic effects, one of the 

### 12 models, I think it was the last one in your table, was talking 

### 13 about the impact of pesticides in sort of a chronic fashion 

### 14 rather than in terms of mortality, on laying of eggs and so on. 

### 15 What was the basis of that model if there is really a lack of 

### 16 data to put that together? 

### 17 **MR. KEITH SAPPINGTON:**Are you referring to 

### 18 the Thompson model? 

### 19 **DR. PAUL SCHWAB:**Yes I am. 

### 20 **MR. KEITH SAPPINGTON:**That was built on - in 

### 21 that paper, the authors actually coupled semi-field experiments 

### 22 on colonies and used that information as input into their -- it 

### 23 was actually a colony model or, excuse me, a model developed for 

### 24 Varroa mites, actually predicting Varroa mite infestations. 

### 25 They adapted that model, which was published earlier, and then 

### 26 included information from their colony-level tests as well as -

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1 I think, I'll have to check - individual level information from 

### 2 various pesticides, and then looked at the results from that, 

### 3 both in terms of the empirical studies as well as the 

### 4 simulation. 

### 5 **DR. PAUL SCHWAB:**So, the effect of the 

### 6 pesticide was a negative impact on the bees from the pesticides 

### 7 themselves, or somehow the pesticides interacting to take care 

### 8 of the mites that they were originally looking at? 

### 9 **MR. KEITH SAPPINGTON:**Keith Sappington. They 

### 10 did not include mites in this particular simulation. The model 

### 11 itself was derived from a model built to investigate mite 

### 12 infestations, but they adapted that and then coupled measured 

### 13 data on colony level effects, and basically tried to compare the 

### 14 two. 

### 15 **DR. DANIEL SCHLENK:**Okay. Dr. McManaman? 

### 16 **DR. JAMES MCMANAMAN:**Yeah, I wonder if we 

### 17 could bring up Mr. Moriarity's presentation. I have a question 

### 18 that's related to models, but it's more related to population of 

### 19 bees. Is that possible? 

### 20 **DR. DANIEL SCHLENK:**They are working on it. 

### 21 Is there another question that we can do while they're bringing 

### 22 that up? Does somebody have something else that we can do? Oh, 

### 23 he's got it. 

### 24 **DR. JAMES MCMANAMAN:**It's number four. So, I 

### 25 was struck by this, and I didn't know exactly - it didn't seem 

### 26 like it was an appropriate time to ask during the presentation.

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1 2 This is kind of an unusual curve in that it 

### 3 looks like there is a steady decline from about 1950 to about 

### 4 1970 in the pollinator population. Then, there's this plateau, 

### 5 kind of up and down plateau for about 20 years, and then when 

### 6 the mites hit, there is a sharp decline. Then, from you know, 

### 7 1995 to about 2006, it looks like there that there is another 

### 8 plateau here. 

### 9 So, this looks like there is a multivariable 

### 10 interaction going on with the pollinator population in general. 

### 11 Has there been any kind of analysis to try to attempt to look at 

### 12 potential -- kind of like an epidemiological analysis -- to look 

### 13 at what was going on? Because in order to understand what could 

### 14 contribute to pollinator decline would really be nice to know 

### 15 what environmental changes have happened, what other kinds of 

### 16 factors could have gone into the declines that we have seen 

### 17 historically. 

### 18 And part of this question is that this 

### 19 plateau, the last plateau, it looks like there is a bit of a 

### 20 decline at about 2005/2006 - do you have more recent data on 

### 21 that the know does that continue to go down or is that just kind 

### 22 of a blip and it really steadies off? 

### 23 **DR. THOMAS STEEGER:**This is Tom Steeger. A 

### 24 couple of comments about this graph. I think that you need to 

### 25 be careful in terms of interpreting it as it speaks to 

### 26 pollinator declines per se. It's certainly informative about

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1 the issue of pollinator declines. This is a graph of the number 

### 2 of commercial colonies that are used in honey production in the 

### 3 United States. 

### 4 What it is probably more reflective of is a 

### 5 changing demographic; that after the war there wasn't the need 

### 6 for people to produce sweetener because sugar was in short 

### 7 supply. And so, more and more people got out of the business of 

### 8 honey production and the other part of this, it's hard to say 

### 9 how much of an impact it had, was on the way that NASS actually 

### 10 collected data, and their exclusion of sort of back yard 

### 11 beekeepers that are producing honey to more commercial honey 

### 12 production is an artifact that is part of this graph. So, it 

### 13 doesn't really present what might be represented -- the 

### 14 contribution from actual pollinator declines. So you have to be 

### 15 careful with how you interpret this graph. 

### 16 As was pointed out in the presentation, it's 

### 17 not - this graph goes out to 2006. The peak was about roughly 6 

### 18 million colonies. It was actually 5.5 million in the late 

### 19 1940s. The most current estimate, the 2011 estimate, places the 

### 20 number of managed honey bee colonies for honey production at 

### 21 2.49 million. So, you still see this decline taking place. As 

### 22 was discussed in Tom Moriarity's presentation, these losses that 

### 23 are being reported by beekeepers of 30 to 34 percent and the 

### 24 high 20s this year because of a mild winter, these are real 

### 25 losses. It's not just changing demographics. 

### 26 There is something happening that is really

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1 difficult to understand in terms of what combination of factors 

### 2 are actually causing it. But getting to your question about 

### 3 epidemiological approaches, it's interesting you should bring 

### 4 that up because Rosalind James certainly has proposed that to 

### 5 potentially pull in super-computing capabilities to get at that 

### 6 issue. It's certainly an issue that USDA has been tasked with 

### 7 over the past five years, congressionally mandated to determine 

### 8 causes, and the week after next, there is actually a conference 

### 9 to look at causal analysis on what is causing bee decline. 

### 10 So, there is a lot of interest and energy 

### 11 being put at addressing this. It certainly was an issue that 

### 12 was flagged by NAS in their publication on pollinator declines 

### 13 in North America, but relative to this graph in terms of 

### 14 interpreting plateaus and the peaks, it can be difficult to 

### 15 interpret. It looks like Dr. Berenbaum has something to add. 

### 16 **DR. MAY BERENBAUM:**Yeah, this was a subject 

### 17 of discussion of the NRC, a study on the status of pollinators 

### 18 in North America. Just to amplify what Dr. Steeger said, this 

### 19 is only, it's not pollinator decline. It's honey producer 

### 20 decline. These data don't reflect the changing nature of the 

### 21 apiculture industry with a greater emphasis on provision of 

### 22 pollinator services. It's just honey production up through 

### 23 2006. That steep drop in the mid 1980s also, although it's easy 

### 24 to attribute that drop to the introduction of mites, it 

### 25 coincides with the change in methodology. So, there is a 

### 26 confounding factor there.

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1 As you said, the sideliners, hobbyists, anyone 

### 2 with five or fewer hives is not included. So, it's not suitable 

### 3 for epidemiological analysis. It's really reflective more of 

### 4 the industry than it is of the pollinator workforce of the 

### 5 United States. You know, it's hard to over interpret because it 

### 6 was not initially set up for that purpose. It was more for 

### 7 honey production. 

### 8 **DR. JAMES MCMANAMAN:**This is Jim McManaman 

### 9 again. So, what is the state and in terms of assessing the role 

### 10 of pesticides and their contributions to decline, whether this 

### 11 is through honey production, but in terms of overall pollinator 

### 12 decline, it would be helpful to really have an accurate idea of 

### 13 what the real nature of the decline has been because it sounds 

### 14 like this is maybe an artificial - it may not be reflecting 

### 15 reality. So that's the basis of my question. Is it still going 

### 16 down or has it plateaued off? 

### 17 **DR. MAY BERENBAUM:**Maybe Jeff, Dr. Pettis 

### 18 could address the Apiary Inspector of America Effort to obtain 

### 19 better data. 

### 20 **DR. DANIEL SCHLENK:**Go ahead, Jeff. Thanks. 

### 21 **DR. JEFF PETTIS:**Very briefly, Jeff Pettis, 

### 22 USDA. As May pointed out, this is just managed honey bee, and 

### 23 actually there has been an analysis of the slope before that 

### 24 blip and afterwards that it's the same - it's mostly driven by 

### 25 economics, honey prices in fact. 

### 26 As far as getting at the loss rate of let's

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1 say honey bee colonies in the U.S., NASS was encouraged to take 

### 2 over and do a very large study and they had no funding to do 

### 3 that. There has been an effort for six years now, but we've 

### 4 done a survey of losses in the U.S. and we asked a number of 

### 5 questions. 

### 6 We're looking at ways to probe that data and 

### 7 try to get at, you know, Varroa mites versus pesticide exposure, 

### 8 et cetera. It's not the best dataset, but it's the best dataset 

### 9 we have. We are looking at various ways to use that dataset to 

### 10 try to get at some of these multifactorial issues, but a lot of 

### 11 it is based on questionnaires where we have, depending on the 

### 12 beekeeper to give us truthful answers. 

### 13 So, we have some other better datasets, but 

### 14 only for specific colonies, two to three hundred colonies, not a 

### 15 huge dataset. 

### 16 **DR. DANIEL SCHLENK:**Okay. Any questions for 

### 17 the Agency? Thanks. Any questions? Dr. McManaman? 

### 18 **DR. JAMES MCMANAMAN:**This is Jim McManaman 

### 19 again. So, I'm going to push this point just a little bit more. 

### 20 So, in terms of understanding the risk and developing risk 

### 21 assessment, it really would be good to have numbers about if 

### 22 there are any inaccuracies and what the actual decline is, it 

### 23 really would in terms of setting policy and going forward on 

### 24 this, it doesn't sound like we really have the numbers in hand 

### 25 to really assess the real risk to the pollinators. Maybe I'm 

### 26 over stating that.

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1 **DR. THOMAS STEEGER:**This is Tom Steeger. I 

### 2 think that if there's any uncertainty in the room that 

### 3 pollinators, particularly the honey bees, where there is a 

### 4 question as to whether they are experiencing significant 

### 5 declines, as someone who is responsible for monitoring the 

### 6 incident database, I have received calls and talked to 

### 7 individual beekeepers that have lost within a period of two 

### 8 weeks 21,000 colonies. As a biological event, if you figure 

### 9 that there are 50,000 bees per colony, the loss of 21,000 

### 10 colonies in that short of a period of time is a staggering 

### 11 event. 

### 12 We are unclear as the role that pesticides are 

### 13 playing in pollinator declines. There have been multiple 

### 14 factors as was discussed in Tom Moriarity's presentation that 

### 15 USDA has identified as contributing to pollinator declines. Do 

### 16 we have a firm understanding of what declines are actually out 

### 17 there? I don't think so, because it's difficult to get a handle 

### 18 on this data, but I think that as an agency that is responsible 

### 19 for regulating pesticides, we have a responsibility to document 

### 20 to the best of our extent, the extent that pesticides are 

### 21 playing a role and to mitigate those effects to the extent 

### 22 possible. 

### 23 But it has been my experience based on 

### 24 information that I get through incident reports and through my 

### 25 interactions with the beekeeping community in the United States 

### 26 that these losses are significant, and people who have been in

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1 the business of beekeeping who have an understanding of how to 

### 2 keep bees are experiencing staggering losses. This is not a 

### 3 contrivance. It is a real problem. The ability of the country 

### 4 to meet pollination service requirements is becoming a question, 

### 5 given the increased need for pollination services for 

### 6 pollinator-dependent crops. 

### 7 **DR. JAMES MCMANAMAN:**This is Jim McManaman 

### 8 again. I certainly didn't want to imply that there was a 

### 9 contrivance, but you know, as a matter of it's just difficult to 

### 10 assess what the contributions of pesticides might be to the 

### 11 loss. There may be multiple factors leading to it. So, without 

### 12 hard numbers, it's just really, you know, we might be shooting 

### 13 at the wrong target is all I'm saying. 

### 14 **DR. DANIEL SCHLENK:**Yeah, Dr. Fefferman? 

### 15 **DR. NINA FEFFERMAN:**Something occurred to me 

### 16 very briefly. For the transparency and clarity aspects of the 

### 17 demands from the modeling side, transparent and clear to whom? 

### 18 To practitioners? To biologists? To beekeepers? To modelers? 

### 19 Where does that benchmark get set? 

### 20 **DR. THOMAS STEEGER:**Presumably to the risk 

### 21 manager as the primary client of our risk assessment, but it's 

### 22 to the stakeholders as well who are affected by our decisions. 

### 23 **DR. NINA FEFFERMAN:**So does that mean that 

### 24 the methodology by which it was arrived must also be clear 

### 25 because I think that might eliminate almost all modeling from 

### 26 being sufficiently transparent.

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1 **DR. THOMAS STEEGER:**As Keith Sappington 

### 2 pointed out, the Agency has specific guidance on the development 

### 3 of models and the transparency has to do with the coding that 

### 4 goes into the algorithms that are used to support those models, 

### 5 and by the testing that goes on subsequent to determine how well 

### 6 those model estimates reflect actual exposure or effects that 

### 7 occur at the colony level. 

### 8 **MR. KEITH SAPPINGTON:**Keith Sappington. I 

### 9 would add in addition to just having the equations in the Paper, 

### 10 having the basis for parameters that are used for those 

### 11 equations, and then again, having that information. So, then it 

### 12 could be evaluated by anybody in terms of a scientific 

### 13 defensibility. 

### 14 **DR. NINA FEFFERMAN:**So, thank you. I mean I 

### 15 guess part of why I'm asking is that again, moving towards the 

### 16 more analytic side of models. There are occasionally very 

### 17 valuable insights that can be provided in the absence of data or 

### 18 validation in the sense of ground truthing outcomes, as long as 

### 19 the process itself is understood to be reflective of a realistic 

### 20 process. So, in some sense, there are no parameters that come 

### 21 in, there are simply functional relationships. And in that 

### 22 sense, then the equations can still provide quite a bit of 

### 23 insight into things like the existence of a meaningful threshold 

### 24 within, for example, this thing can't go negative, and therefore 

### 25 there exists no satisfiable threshold, because otherwise we've 

### 26 created bees out of thin air kind of validity.

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1 On that level, sometimes there can be English 

### 2 descriptions, but there isn't so much to grab on to. So, I'm 

### 3 wondering are there guidelines that apply equally well to models 

### 4 like that? Are they necessarily then off the table? What 

### 5 happens? 

### 6 **DR. STEPHEN BRADBURY:**Maybe there is another 

### 7 document we could try to get into the record, which may get a 

### 8 bit at what you're asking. There is an OECD document, and it's 

### 9 for quantitative structure activity relationships, which is a 

### 10 different specific topic, but it talks about model - it calls it 

### 11 model validation; but it is more around what kind of thing 

### 12 should you kind of describe about your model. 

### 13 It doesn't come at that a model is right or 

### 14 wrong, it's about describing what the model does and various 

### 15 attributes of the model and explaining it in such a way that a 

### 16 user of the model can be informed as to whether or not it's 

### 17 appropriate to use a given model for the given purpose that 

### 18 somebody maybe - or question somebody may be trying to answer. 

### 19 So, I don't know if it will match exactly what 

### 20 you're getting at, but I think it might provide some insights 

### 21 into where we're coming from. It isn't the model is good, bad 

### 22 or indifferent, it's how do you try to describe attributes of 

### 23 the model so that you can understand how to use it or not use it 

### 24 or use it with care for a given question you may need to answer. 

### 25 26 Getting back to your transparency and

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1 understandability, certainly it's in the risk assessment 

### 2 framework that the risk assessor and the risk manager hopefully 

### 3 understand each other so they're not passing in the night and 

### 4 burning resources in time, misunderstanding each other; but also 

### 5 at the end of the day it's to ensure that the public understands 

### 6 what it is we're thinking and how we formulated out position. 

### 7 Clearly, sometimes these documents are huge. So, we have a 

### 8 range of audiences from scientific experts that know the very 

### 9 excruciating detail of a certain component of the risk 

### 10 assessment and somewhere in that risk assessment document, being 

### 11 able to explain it to that audience. But ultimately being able 

### 12 to go through risk communication and analysis and writing so 

### 13 that public can understand the rationale to the decision making. 

### 14 So it gets challenging, but that's what we're trying to strive 

### 15 for. 

### 16 **DR. NINA FEFFERMAN:**Thank you, that sounds as 

### 17 though the extra document you were thinking of would get to 

### 18 exactly the sort of thing that I'm thinking, and it does also 

### 19 set a useful benchmark for me in considering. Thank you. 

### 20 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 21 **DR. JEFF PETTIS:**More of a comment, but it 

### 22 does tie into the modeling, and it gets back to your question a 

### 23 bit. 

### 24 As you go from Tier I to Tier II or Tier III, 

### 25 you go from control studies with individual bees, which we can 

### 26 control. We can control the genetics. We can control

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1 everything. Then you get an interpretation of some result of 

### 2 these individual bees. As soon as you take it Tier II or Tier 

### 3 III, where you have colony level, you bring in all this 

### 4 uncertainty. I think that's what we grapple with. 

### 5 I mean, so maybe a modeling approach, 

### 6 especially at the colony level, it can be very useful and maybe 

### 7 applying a certain degree of loss or something attributable to 

### 8 one thing and then following that over time. But as soon as you 

### 9 go to the colony level, it's impossible to control all these 

### 10 other factors, and that's what we grapple with. 

### 11 **DR. DANIEL SCHLENK:**That would be a great 

### 12 discussion for tomorrow and the next day. Looking for any other 

### 13 questions for the Agency while we got them here. Yes, Dr. 

### 14 Potter? 

### 15 **DR. THOMAS POTTER:**This is Tom Potter here. 

### 16 We've been looking at question 14 because I was identified as an 

### 17 associate discussant on there. So, I have been trying to get my 

### 18 head around it. I think it's question 14 and it has to do with 

### 19 a colony-level modeling. I though the answer was easy. Yes, 

### 20 it's a great thing. Boy, I wish we had the simulation model 

### 21 where we could assess these effects, but apparently it doesn't 

### 22 exist and they exist in various, what appear to be crude form. 

### 23 So, ultimately my question comes around to you 

### 24 folks is you know, what kind of response are you looking for? 

### 25 Is the answer yes, models are a great idea? I think that one's 

### 26 easy. And the second part of it is who is going to develop

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1 those models and support them? Obviously, that's a 

### 2 fundamentally important question with regard to any simulation 

### 3 model that we might use. 

### 4 **MR. KEITH SAPPINGTON:**Keith Sappington. I 

### 5 think the first thing we wanted was the simple question, perhaps 

### 6 yes, was that is this concept worth dedicating some amount of 

### 7 resources to, because it is a rather large effort even to take 

### 8 an existing model and understand the transparency, understand 

### 9 all of the information that went into it, develop it and modify 

### 10 it and then bring it to a panel like this for final review. 

### 11 So, before doing that and in the context of 

### 12 pollinator declines and multiple stressors, we wanted to just 

### 13 check in before considering this option further. 

### 14 The second thing is getting sort of a check in 

### 15 on this state of the science. Do you agree that with the 

### 16 current state of the science, what we were able to do just 

### 17 briefly was to summarize a few of the models. 

### 18 And then thirdly, sort of helping us evaluate 

### 19 or even perhaps dedicate efforts to developing some models, 

### 20 understanding what are the most important factors to include in 

### 21 a model? 

### 22 The HoPoMo model has 66 differential equations 

### 23 within it, and all models are wrong and some models are useful. 

### 24 So, I'm not sure that we need a model with 66 differential 

### 25 equations. So, the advice that would be very helpful to us 

### 26 would be to say, well here's your top tier of things that you

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1 must include in a model to have any chance of getting a 

### 2 realistic result, and then here are some other sort of secondary 

### 3 tiers that gee, if you have the data, this will be useful and it 

### 4 will be useful for this purpose. 

### 5 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 6 **DR. MAY BERENBAUM:**Not to prolong this, but 

### 7 just to follow up and reflect my profound ignorance of how 

### 8 Washington, DC, works. So, we are likely to identify data gaps, 

### 9 but isn't EPA primarily regulatory and whose responsibility is 

### 10 it then to fill those data gaps? Is it kind of who will help me 

### 11 bake the bread scenario. Yeah, how does our response to the 

### 12 charges figure into meeting the EPA's need for certain types of 

### 13 information? 

### 14 **DR. DANIEL SCHLENK:**Dr. Bradbury, I think 

### 15 would answer that one, yeah? 

### 16 **DR. STEPHEN BRADBURY:**I will just be careful 

### 17 on my editorial comment about Washington. So, it gets back to 

### 18 some of the earlier questions in terms of will it go quick. But 

### 19 the big picture, what's the cause of pollinator decline. That's 

### 20 an activity, USDA is leading it and multiple federal agencies 

### 21 are involved in academia. So, as we said at the beginning of 

### 22 today's discussion, we're focusing on the pesticides first in 

### 23 terms of under FIFRA we do take a look at effects of pesticides 

### 24 on nontarget organisms, pollinators are an important component 

### 25 in the ecosystems. They have benefits for agricultural 

### 26 production and the products of the hive, with everything else

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1 that's going on, making sure we're advancing our capability of 

### 2 doing that. 

### 3 Some of our discussion that was in isolation 

### 4 of interactive effects and we talked a little bit about it today 

### 5 and you may have some insights as we go through the charge 

### 6 questions, that perhaps while not directly related to what we do 

### 7 in the pesticide program, your report combined with the SETAC 

### 8 report any other reports, EFSA report, may help spawn research 

### 9 in different organizations in the federal government or 

### 10 internationally. 

### 11 With regard to specific uncertainties that may 

### 12 relate to doing pesticide risk assessment, there is a variety of 

### 13 scenarios that could play out. The Part 158 test guidelines 

### 14 that some of the speakers discussed already has an opening - 

### 15 more than an opening, it has the ability to do different kind of 

### 16 field studies. But those guidelines are written, I think 

### 17 appropriately so, open and not very prescriptive because they 

### 18 are designed to answer specific questions. 

### 19 Some of the feedback potentially from the 

### 20 charge questions could give us insights as to how to tailor, how 

### 21 to better define what the attributes of the semi-field studies 

### 22 or full-field studies, or maybe variations on the laboratory 

### 23 experiments could be to help focus, because our guidelines 

### 24 already give us some flexibility. That could be a way through 

### 25 chemical, by chemical or risk assessment scenario by scenario to 

### 26 get information, but you would be giving us some guidance on how

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1 to be more efficient or effective in focusing the chemical risk 

### 2 assessment specific issues. 

### 3 There could be some feedback that you provide 

### 4 that with the work we're doing with other countries and OECD or 

### 5 in other bilateral multilateral arrangements, there may be 

### 6 advice along with the SETAC Pellston document that may help 

### 7 advance some test guidelines, say at the laboratory level, that 

### 8 we all could use and then work with the regulating committee to 

### 9 go forward. 

### 10 So, I think there is a variety of ways that 

### 11 this could play out. There may be some really pure research 

### 12 questions that we suggest and there are other parts of the 

### 13 federal family, state, international groups that will read your 

### 14 report and use it as well. 

### 15 **DR. DANIEL SCHLENK:**Okay. Are we done? Any 

### 16 other questions? What we will do just to again summarize, 

### 17 mentioned this, this morning in our early morning, I think the 

### 18 Agency will be here tomorrow after our public commenting period. 

### 19 Before we start questions, you'll have another opportunity to 

### 20 ask any questions that come out tonight in your discussions and 

### 21 deliberations, and just mulling over what has been gone over 

### 22 today that you ask one last time again before we actually get 

### 23 into the charge questions. 

### 24 Okay. All right, before I turn it over to 

### 25 Fred, I just want to acknowledge thanks for the Agency's 

### 26 presentations today, and also for the panel if you would please

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1 meet in the conference room after this, after you can do a 

### 2 restroom break, we need to meet and hash some things out before 

### 3 you head back to your rooms. 

### 4 **DR. FRED JENKINS:**Okay. I just have a few 

### 5 things to say that are directed towards the public commenters 

### 6 for tomorrow. If you are giving a public comment that has a 

### 7 power point presentation, please see me as soon as possible, 

### 8 either now or early in the morning before the meeting, 

### 9 preferably no later than 8:30 tomorrow morning so I can load 

### 10 your power point presentation onto the computer behind me. 

### 11 Also, to all the public commenters, please 

### 12 arrive to the meeting on time tomorrow at 9 a.m. We don't want 

### 13 you to miss your opportunity to provide your public comment. 

### 14 Once the public comment period closes, there will be no more 

### 15 opportunity to provide oral public comments at the meeting. 

### 16 As a reminder, if you have not made prior 

### 17 arrangements with me to provide oral public comments, please see 

### 18 me as soon as possible so we can get you signed up as a public 

### 19 commenter and I ask also as a reminder, please limit your 

### 20 comment to five minutes. 

### 21 And the last point I want to make, I ask that 

### 22 all public commenters please keep your oral comment times to the 

### 23 agreed upon time frame. If you've asked for five minutes, 

### 24 please only speak for five minutes, 10 minutes, please only 

### 25 speak for 10 minutes, an hour, please only speak for an hour. 

### 26 We have a whole lot of ground to cover this

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1 week, and we ask all of our public commenters to please help us 

### 2 in covering and accomplishing what we need to do for this 

### 3 meeting. Please stay within your specified timeframe. And 

### 4 that's all I have to say. This meeting is adjourned for today 

### 5 and we look forward to seeing everyone in the morning. Thank 

### 6 you for coming. 

### 7 (WHEREUPON the meeting was adjourned for the 

### 8 day.) 

### 9 **DR. DANIEL SCHLENK:**Okay. So, let's go 

### 10 ahead and just do a real brief introduction around the panel 

### 11 again. Just state your name and affiliations before we get 

### 12 started so everybody can identify themselves. My name is Dan 

### 13 Schlenk. I am professor in the Department of Environmental 

### 14 Sciences, University of California Riverside and I Chair of the 

### 15 SAP. 

### 16 **DR. STEPHEN KLAINE:**My name is Steve Klaine. 

### 17 Good Morning. I am with Clemson University and I am an aquatic 

### 18 ecotoxicologist. 

### 19 **DR. JAMES MCMANAMAN:**Good morning, I am Jim 

### 20 McManaman. I am at the University of Colorado in the Department 

### 21 of Obstetrics and Gynecology. 

### 22 **DR. KENNETH DELCLOS:**Barry Delclos from the 

### 23 Food and Drug Administration's National Center for Toxicological 

### 24 Research. 

### 25 **DR. DAVID TARPY:**Good morning, I'm David 

### 26 Tarpy. I'm at North Carolina State University in the Department

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1 of Entomology. I am a honey bee biologist and the extension 

### 2 apiculturist for North Carolina. 

### 3 **DR. PAUL SCHWAB:**I am Paul Schwab from Texas 

### 4 A&M University. I am an environmental soil chemist. 

### 5 **DR. THOMAS POTTER:**Tom Potter, USDA 

### 6 Agriculture Research Service, Southeast Watershed Laboratory in 

### 7 Tifton, Georgia. I am a pesticide chemist. 

### 8 **MR. JENS PISTORIUS:**Jens Pistorius, Germany, 

### 9 Julius Kuehn Institute. I work with risk assessment for 

### 10 pesticides in honey bees, investigation of bee poisoning 

### 11 incidents and research pesticides in honey bees. 

### 12 **DR. JEFF PETTIS:**Good morning, I am Jeff 

### 13 Pettis, I am with USDA Agricultural Research Service, Bee 

### 14 Research Laboratory here in Beltsville, Maryland, and I am an 

### 15 entomologist. 

### 16 **DR. NANCY OSTIGUY:**Nancy Ostiguy, from 

### 17 Entomology Department at Penn State. 

### 18 **DR. ROSALIND JAMES:**Rosalind James, I am with 

### 19 the USDA Agricultural Research Service. I am director of the 

### 20 Pollinating and Sex Research Laboratory in Logan, Utah. 

### 21 **DR. GREG HUNT:**Good morning, I am Greg Hunt. 

### 22 I am at Purdue University and I study honey bee genomics, 

### 23 behavior and health issues. 

### 24 **DR. NINA FEFFERMAN:**Good morning, I am Nina 

### 25 Fefferman from Rutgers University. I am an applied mathematical 

### 26 modeler of ecology and epidemiological systems.

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1 **DR. MAY BERENBAUM:**I am May Berenbaum, the 

### 2 Profession and Head of Entomology at the University of Illinois 

### 3 at Urbana-Champagne, and I am an insect chemical ecologist. 

### 4 **DR. DANIEL SCHLENK:**Thank you everyone. 

### 5 Before we get started, Fred has some comments he needs to make 

### 6 to our public commenters. 

### 7 **DR. FRED JENKINS:**Good morning everyone and 

### 8 welcome to the second day of our FIFRA SAP on Pollinator Risk 

### 9 Assessment Framework. My name is Fred Jenkins, and I am the 

### 10 Designated Federal Official for the FIFRA SAP. So, I just 

### 11 wanted to briefly say to the public commenters - we have quite a 

### 12 few public commenters this morning. If you have a power point 

### 13 presentation, please see someone on our staff so that we get 

### 14 your presentation loaded onto the computer behind us. Also, if 

### 15 you have any handouts of your comments that you wanted 

### 16 distributed to the Panel, please bring those now to our staff so 

### 17 that we can have those distributed as well. With that said, I 

### 18 am going to turn the floor over to Dr. Daniel Schlenk. 

### 19 **DR. DANIEL SCHLENK:**Thanks Fred. Couple 

### 20 comments -- this morning just speaking of materials - there were 

### 21 a couple papers that were placed in the chairs of some of the 

### 22 panel members. If you have materials that need to be presented 

### 23 to the panel they need to go through the EPA staff. We can't 

### 24 have materials distributed without a formal distribution pattern 

### 25 through the EPA. So again, if you have materials that you need 

### 26 to get to the Panel or you would like the Panel to see, please

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1 do so through the EPA staff here. Okay. With that, we turn it 

### 2 over to the EPA and any sort of follow-up issues you need to 

### 3 present today before we start with the public comments. 

### 4 **DR. DONALD BRADY:**No, thanks very much Dr. 

### 5 Schlenk, I think we are ready to go right to public comments. 

### 6 **DR. DANIEL SCHLENK:**Great, what I would like 

### 7 to do, if you don't mind, is following the public comment period 

### 8 if you guys could come back up, I will ask the panel if they 

### 9 have any last minute questions related to the materials. Great, 

### 10 thanks. 

### 11 Okay. Our first public commenter is CropLife 

### 12 America, and I believe that is going to be Dr. McAllister making 

### 13 that presentation. Again, as Fred had mentioned, please hold to 

### 14 the time that you have been allotted. We have an hour allocated 

### 15 for you guys, for CropLife, if you can please keep to that time 

### 16 that would be great. 

### 17 **DR. RAYMOND MCALLISTER:**Thank you very much. 

### 18 My name is Ray McAllister. I am the Senior Director of 

### 19 Regulatory Policy for CropLife America. CLA is the National 

### 20 Trade Association representing the crop protectin industry in 

### 21 the United States. We would like to express our sincere 

### 22 gratitude to the bee technical workgroup, which has worked very 

### 23 hard over the past month or so to prepare the written comments, 

### 24 which have been distributed to you as well as the oral summary 

### 25 that you will be hearing this morning. I will just making a few 

### 26 introductory remarks, and then we will hear from David Fischer

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1 of Bayer and Jay Overmyer of Syngenta for the bulk of the 

### 2 remarks today. 

### 3 We all recognize that pollinator health is a 

### 4 vital issue for American agriculture. CLA in particular 

### 5 recognizes that crop protection technology, as well as the 

### 6 pollination services provided by both managed honey bee colonies 

### 7 as well as native pollinators are necessary for successful 

### 8 agriculture in feeding a hungry planet. 

### 9 CropLife America commends the efforts of EPA, 

### 10 PMRA and Cal DPR in improving the science of pollinator risk 

### 11 assessment. Our member companies appreciate this opportunity to 

### 12 provide input into a very important process. We believe that 

### 13 improving the quality and rigor of risk assessments will ensure 

### 14 that the crop protection tools, which are critical for 

### 15 sustainable agriculture production, can be used in a manner 

### 16 compatible with pollinator health. 

### 17 Pollinator health can be adversely affected by 

### 18 a combination of stress factors, which include a variety of 

### 19 diseases, some very devastating parasites, problem with 

### 20 nutrition, loss of habitat, chemical stress and bee management 

### 21 practices. 

### 22 There are some who have asserted that 

### 23 pesticides are the root cause of colony collapse disorder in 

### 24 honey bees, but this is not supported by the weight of the 

### 25 evidence. We believe that establishment of a scientifically 

### 26 credible risk assessment approach for pollinators will help

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1 promote sustainable agricultural practices and these are of 

### 2 benefit to all involved, from farmers, beekeepers and the 

### 3 American Consumer. 

### 4 The establishment of sound and scientifically 

### 5 valid test methods and risk assessment processes should aid all 

### 6 interested researchers in optimizing study designs to ensure 

### 7 that the results are reproducible and that they can be 

### 8 interpreted clearly. Lack of such guidance has led to 

### 9 publication of some recent studies, which are not optimized for 

### 10 contribution to risk assessment and have led to some erroneous 

### 11 conclusions about the relative importance of pesticides in 

### 12 pollinator health in practical circumstances. 

### 13 The improved risk assessments will be 

### 14 important to support the appropriate labeling of pesticide 

### 15 products and accompanying best management practices for 

### 16 safeguarding pollinator health. 

### 17 CLA believes that the scientifically sound 

### 18 risk assessment procedures are critical not only to ensure that 

### 19 the EPA's and the nation's environmental protection goals are 

### 20 met, but to instill confidence in the public in the regulatory 

### 21 approval process. In the hopes of improving these risk 

### 22 assessment processes under review, we are offering the following 

### 23 comments, and I will turn the time over to Dave Fischer. 

### 24 **DR. DAVID FISCHER:**Thanks Ray. I am David 

### 25 Fischer. I am an ecotoxicologist with Bayer CropScience, and on 

### 26 really the comments that I am presenting and Jay Overmyer from

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1 Syngenta will be presenting is the work of - technical workgroup 

### 2 we have among the companies and CropLife America. 

### 3 First, some general comments on the White 

### 4 Paper Risk Assessment Process. We support the harmonized 

### 5 approach among the three regulatory agencies. This is something 

### 6 that is of great benefit to our industry that we have a 

### 7 consistent approach stake in various jurisdictions. Not only in 

### 8 North America, but we would like to see harmonization around the 

### 9 world as well. 

### 10 The tiered risk assessment approach we 

### 11 support. This is similar to what we do with other taxa. We try 

### 12 and screen out the chemicals that you can eliminate concern 

### 13 about, focus the resources where there is the main 

### 14 uncertainties; and we also support expanding the Tier I data 

### 15 requirements as the Pellston Workshop recommended and as the 

### 16 White Paper recommends to fill in some of the gaps we currently 

### 17 have with just the limited tests that are in the current 

### 18 guidelines. 

### 19 We also support a flexible approach to doing 

### 20 higher tier testing, Tier II and Tier III. It would be nice if 

### 21 there was a prescription for doing this, but every chemical in a 

### 22 use pattern really will have its own differences that need to be 

### 23 taken into account. The key thing is to make sure that when we 

### 24 go to higher tiers, we are focusing on what the uncertainties 

### 25 are and addressing those with the studies. 

### 26 We do have some concerns. We think the Tier I

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1 assessment, especially the way the dietary exposure is being 

### 2 estimated, is overly conservative and will lead a lot of 

### 3 chemicals that are reasonably safe or really should not cause a 

### 4 concern progressing to higher tiers. I will say more details 

### 5 about that, why we think that might be true; and also how we 

### 6 think it might be remedied. 

### 7 We are also concerned if we get into a system 

### 8 where a lot of chemicals have to go to higher tier testing that 

### 9 we have clear acceptability criteria in place for those higher 

### 10 tier tests and how they are going to be conducted, how they the 

### 11 data is going to be interpreted. Right now, there is a lot of 

### 12 uncertainty there. 

### 13 So, we will go through sort of the parts of 

### 14 the White Paper with respect to problem formulation. We support 

### 15 the protection goals, which largely were the same as we 

### 16 developed at the Pellston Workshop, which our industry 

### 17 participated in along with the EPA scientists and academic 

### 18 scientists. 

### 19 One thing that is appreciated in the White 

### 20 Paper that we have not always seen in other assessment documents 

### 21 is a real clear explanation for how levels of concern are 

### 22 established. We appreciate the clarity and the transparency of 

### 23 how they said this is what we are trying to do and this is the 

### 24 RQ value that seems to meet that criteria; and its science 

### 25 based, so we support that. 

### 26 I mentioned flexible testing beyond Tier I. I

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1 won't say anything more about that. Use of conceptual model - 

### 2 we thought that this White Paper has a very good illustration of 

### 3 how to do a conceptual model and how to use it to focus the 

### 4 assessment and the development of higher tier data to address 

### 5 the main uncertainties. It is very difficult to assess 

### 6 everything in an assessment, and I think this does a good job of 

### 7 focusing on that major routes of exposure. 

### 8 Also, we support using honey bees as a 

### 9 practical surrogate for pollinators in general. This was 

### 10 discussed at length at the Pellston Workshop and the report will 

### 11 have some more information on non-Apis bees, but for practical 

### 12 purposes and certainly with the effects testing, the honey bee 

### 13 should be a reasonable surrogate. 

### 14 Okay. Characterization of Contact Exposure - 

### 15 we basically support what is proposed in the White Paper using 

### 16 the relationship from the Koch and Weisser Database. It seems 

### 17 to be the best available data. It's consistent with what was 

### 18 agreed to at the Pellston Workshop. As the White Paper analysis 

### 19 shows, it seems to be an improvement on previous approaches. 

### 20 Now into some areas where we see some room for 

### 21 improvement. Characterization of Dietary Exposure - first of 

### 22 all, the soil applications, the Briggs' model, we actually think 

### 23 is reasonably good Tier I starting point. You know, if you have 

### 24 to come up with an estimate of systemic uptake into plants, into 

### 25 field crops, this is a reasonable starting point. It doesn't 

### 26 necessarily tell you what is going to be expressed in the

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1 flowers of a plant. It is not developed to do that, but it 

### 2 should be, if anything, an overestimate of that. 

### 3 Again, as more data are developed, this will 

### 4 be a theme on a lot of our slides. Some of these Tier I 

### 5 estimates may need to be reevaluated. 

### 6 With respect to seed treatments, the default 

### 7 value of PPM or mg/kg, I think was originally proposed for all 

### 8 systemic risk assessments. So, whether it was a seed treatment 

### 9 or a soil treatment, that seemed to be an upper-bound for seed 

### 10 treatments. However, the field data are quite a bit less. If 

### 11 you look at the field measurements in table 17, the highest 

### 12 value is 36 parts per billion and the Tier I estimate is more 

### 13 than an order of magnitude; it's like 30 times higher than that. 

### 14 It looks to us like a default value of 0.1 mg/kg would be more 

### 15 appropriate and adequately conservative just based upon the data 

### 16 that we have. 

### 17 For soil treatments, the numbers can be 

### 18 higher, but when they are the Briggs' model does an adequate job 

### 19 of reflecting that. 

### 20 Again, I think maybe it's implied in the White 

### 21 Paper, but it doesn't specifically state, so we want to state 

### 22 it. There is no point in concluding there is a risk of use of 

### 23 seed treatments that aren't systemically active. There are a 

### 24 lot of them; especially a lot of the fungicides won't 

### 25 necessarily be expressed in the pollen and nectar to any extent. 

### 26 We should be able to screen those out rather than just assume

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1 that there is a concentration there. 

### 2 Tree trunk applications - here is an area 

### 3 where we are in need of a lot more data, a lot of research. So, 

### 4 what the Agency has come up with is a back-of-the-envelope 

### 5 calculation that basically tries to say what if all the mass of 

### 6 compound applied is evenly distributed in the biomass of the 

### 7 tree, the leaves and flowers. There is a lot of uncertainty 

### 8 whether this is an accurate prediction. We think it is probably 

### 9 very conservative because flower concentrations are often orders 

### 10 of magnitude less than leaf concentrations, but it is a starting 

### 11 point. 

### 12 So, you know, we think it is going to be very 

### 13 conservative. We do think that there may be some data out there 

### 14 from the USDA studies on Asian longhorn beetle control programs, 

### 15 Emerald Ash borer, maybe the Hemlock Woolly Adelgid. I think - 

### 16 and we don't have access to all these data, but there may be 

### 17 some data that USDA can pull in here that EPA hasn't considered 

### 18 yet. 

### 19 Okay. So, our main concern is the foliar use, 

### 20 the dietary estimate that is being proposed is basically to use 

### 21 the tall grass value from the T-REX model, which really comes 

### 22 from the Hoeger and Kenaga nomogram as modified by Fletcher et 

### 23 al. For just looking at the numbers versus what these estimates 

### 24 are, the pollen numbers appear to be reasonably conservative. 

### 25 They're high. They're about 2X higher than the highest value 

### 26 that is in the dataset, but the dataset is small, so that

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1 doesn't seem to be unreasonable as a starting point. 

### 2 But for nectar, it's much, much higher. It 

### 3 appears to be overly conservative with the highest field 

### 4 measurements more than an order of two magnitude below what the 

### 5 T-REX model value is. 

### 6 So, since nectar consumption, nectar 

### 7 concentrations really are what drive this risk assessment -- if 

### 8 you crunch the numbers 97 percent of the exposure comes from 

### 9 nectar and 3 percent from pollen according to the calculations 

### 10 -- it's critical that you not grossly overestimate the nectar 

### 11 concentrations. 

### 12 So, just looking at the data, and I think I 

### 13 will transition to the next slide, looking at the data in table 

### 14 5, it looked to us that there is about a 3X factor between 

### 15 pollen and nectar, particularly when you look at the studies 

### 16 that are reporting average field measurements. We put more 

### 17 weight on those than the maximum ones because those can be just 

### 18 more variable. 

### 19 So, we are proposing that you take the value 

### 20 for pollen and multiply it by 0.33 to get the estimate for 

### 21 nectar. 

### 22 Then the question - how conservation should 

### 23 Tier I exposure estimates be? There is a statement that the 

### 24 Tier I method is intended to generate reasonably conservative 

### 25 estimates. We agree with that; but meaning that the estimates 

### 26 should generally be within one or two orders of magnitude higher

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1 than the true environmental exposure. That seems to us to be 

### 2 too high. One to two orders of magnitude bias in your estimate 

### 3 can greatly change the number of the compounds that you fail to 

### 4 screen out and have to conduct higher tests with. 

### 5 If you look at what Fletcher did in developing 

### 6 the nomogram values that are used in T-REX, they look at field 

### 7 residue measures and just took the 95th percentile. Really, if 

### 8 you look at EPA policy in the framework documents, they refer to 

### 9 high end estimates usually being in the 90th-95th percentile 

### 10 range; that is traditionally what is done. Now here we do not 

### 11 have a lot of data, so we may not have enough to just say use a 

### 12 95th percentile, but taking the highest value that you have and 

### 13 going 10X or 100X more than that seems to us to be too 

### 14 conservative. 

### 15 We point out that the distributions and 

### 16 residue measurements tend to be log-normal, which means they are 

### 17 skewed to the low side with a fewer high-end values. Those 

### 18 high-end values can be statistical outliers and probably are 

### 19 less reliable. Sometimes, they may even be errors due to 

### 20 contamination when the sample is collected or analytic 

### 21 methodology. 

### 22 Just what is the consequence if you over 

### 23 estimate exposure by 10 to 100 times? Just as an example, a lot 

### 24 of us probably had our morning coffee. If you look at caffeine, 

### 25 I don't know if you know this, but caffeine is actually pretty 

### 26 toxic.

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1 The LD50 is in the range of 150-200 mg/kg body 

### 2 weight, which is about 80 cups of coffee with a typical strength 

### 3 of caffeine. So, that's about a little over 1 percent of an 

### 4 LD50 in each cup of coffee. 

### 5 If you over estimate the caffeine 

### 6 concentration in coffee by 10X, you conclude that each cup has 

### 7 more than 10 percent of an LD50 dose, which in most risk 

### 8 assessment schemes would at least cause some concern. If you 

### 9 overestimate it by 100X, you can conclude that a cup of coffee 

### 10 is lethal to a lot of people. So clearly that would -- you know 

### 11 just as an analogy -- I think we would want to screen out very 

### 12 reasonably safe products like we would screen out having to do 

### 13 high risk assessments for caffeine in coffee. 

### 14 The other thing is the food intake rate, which 

### 15 I think there is a lot of uncertainty, or at least we are 

### 16 wondering if that needs to be looked at - and I really 

### 17 appreciate what the Agency did because they pulled off a higher 

### 18 tier methodology approach by using a Monte Carlo procedure to 

### 19 try and take the data that Rortais et all had and come up with 

### 20 an estimate of how much food a bee ingests in a day based upon 

### 21 energetics and other things. We did a lot of the similar things 

### 22 in ECOFRAM a while back. Having done some of that modeling, 

### 23 some of the pitfalls were made aware to me because I made some 

### 24 of the same mistakes. 

### 25 First of all, log-normal distribution was 

### 26 assumed for the sugar concentration in nectar. That might be

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1 fine, but just setting the mean at 30 percent means that most of 

### 2 those trips out there, the assumption is that they're coming 

### 3 back with nectar with less than 30 percent because the 

### 4 log-normal is going to be skewed to the low side. 

### 5 Of course, why is that significant? It's 

### 6 because you are basically totaling up the amount of nectar the 

### 7 bees are ingesting in order to get a certain amount of sugar and 

### 8 summing the mass of pesticide in that quantity of nectar. So, 

### 9 there could be some bias there. At least it is something that 

### 10 you might want to look at. 

### 11 There is also an assumption that bees are 

### 12 foraging randomly. They don't show any preference for picking 

### 13 out food sources that have higher sugar content. I wonder if 

### 14 that is true. I would be surprised if it was. I think if you 

### 15 put a bunch of feeders out there, and there are probably people 

### 16 on the panel that know the answer to this, you would find that 

### 17 they would select the higher sugar source. You know, it would 

### 18 just be -- optimal foraging theory would predict they would. 

### 19 There are three variables in this Monte Carlo 

### 20 Simulation that use uniform distribution. This is where some of 

### 21 my personal experience comes in. I did this myself. I put in 

### 22 uniform distributions in some of the ECOFRAM models and when 

### 23 Dwayne Moore reviewed them he told me, don't ever put in a 

### 24 uniform distribution. It puts too much weight on the tails, 

### 25 come up with some sort of beta distribution that uses expert 

### 26 judgement.

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1 So, I don't know if this changes - you know, I 

### 2 would want to see a sensitivity analysis to see if changing the 

### 3 distribution would change the result of the simulation. 

### 4 Then finally, number of foraging flights per 

### 5 day - I think and I could be wrong, but there was nothing that 

### 6 indicated to me when I read it that they weren't just assuming 

### 7 this was an independent variable from the distance traveled or 

### 8 the duration of the flight. It makes sense to me that wouldn't 

### 9 you expect this to be highly correlated that if you were making 

### 10 more flights per day, they were shorter flights. So, you 

### 11 wouldn't need as much energy, you wouldn't need as much sugar to 

### 12 fuel those flights. 

### 13 So, if you have correlated variables - and 

### 14 there are ways of doing that - you want to specify that in your 

### 15 modeling. I'm not sure if that was done. All of these factors 

### 16 may result in a nectar consumption being over estimated. 

### 17 So, I want to support the concept of using 

### 18 this sort of modeling technique. I just think it might need to 

### 19 be vetted. 

### 20 Dose from food intake - another thing is and 

### 21 this is common to assessments we do for birds and mammals as 

### 22 well - we do back-of-the-envelope, you know how many LD50s per 

### 23 day does an animal get? That's kind of what we are doing. We 

### 24 are calculating the cumulative dose over entire day of foraging 

### 25 per bee, but we are not taking into account metabolism and 

### 26 detoxification that is going on as this food is being ingested.

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1 That can be a significant factor. 

### 2 Going back to the caffeine example with 

### 3 coffee, you get a different dose if you have one cup of coffee 

### 4 an hour versus if you have 10 at once. You know, the caffeine 

### 5 is metabolized and detoxified, excreted so the doses don't 

### 6 necessarily add up, aren't completely additive. So, that is 

### 7 another source of conservatism in the assessment. 

### 8 Then there are a lot of other factors. We 

### 9 support the option of addressing uncertainties in the risk 

### 10 assessment factors with whole colony exposure studies, which 

### 11 really is, I think, where we need to get better data and really 

### 12 find out what the colony is being exposed to. 

### 13 Then there is in these other bullets, all 

### 14 these other factors that need to be taken into account as far as 

### 15 the use patterns and just how likely is exposure to occur. We 

### 16 would hate to see us wasting time doing higher tier assessments 

### 17 on exposure scenarios that should be screened out just on the 

### 18 basis that the way you are using a product as a bait or applying 

### 19 to a crop that bees aren't going to visit like leaf lettuce or 

### 20 applying post bloom. All of these factors I think EPA currently 

### 21 takes into account. We just want to emphasize that maybe the 

### 22 White Paper did not state that as clearly as needs to be stated. 

### 23 So there will be other uses that will be screened out besides 

### 24 just based upon risk quotients. 

### 25 Okay. I will turn the next piece on effects 

### 26 and risk assessment over the Jay Overmyer.

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1 **DR. JAY OVERMYER:**Thanks Dave. Again, my 

### 2 name is Jay Overmyer. I am an ecotoxicologist with Syngenta 

### 3 Crop Protection. I am going to go over a couple of the slides 

### 4 here related to effects testing. 

### 5 As Dave mentioned earlier, CLA supports the 

### 6 use of tier testing and the approach in the risk assessment. 

### 7 Obviously moving from Tier I to Tier III, if looking at the 

### 8 amount of ability to control the situation or control the test, 

### 9 there is a reduction in the amount of control from Tier I to 

### 10 Tier III. Also, moving from Tier I to Tier III, we also have an 

### 11 increase in environmental realism within those studies. 

### 12 So the Tier I studies are highly controlled 

### 13 laboratory tests designed to predict an LD50 or NOAEC, often 

### 14 subjecting the organisms to high doses to achieve those 

### 15 endpoints. Basically to come up with these endpoints, we are 

### 16 able to determine what the relative toxicities of these 

### 17 compounds are. 

### 18 Moving to Tier II, again a little bit less 

### 19 controlled, semi-field type studies. Exposure is closer to real 

### 20 world, but yet we are still looking at forced exposures with the 

### 21 bees inside of a tunnel or an enclosure, which inhibits them 

### 22 from naturally foraging that they might do. The benefits of the 

### 23 Tier II we're also increasing environmental realism, therefore 

### 24 we're getting up to a hive-level effect as opposed to 

### 25 individual-level effect. 

### 26 Then with Tier III, obviously again, we're

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1 losing a little bit more control. The bees are allowed to 

### 2 forage naturally, often looking at natural application rates and 

### 3 exposures, and also being able to obtain information related to 

### 4 the hive, which is important. 

### 5 For the Tier I testing, CLA does support that 

### 6 the use of the Tier I test, the adult contact and oral acute 

### 7 toxicity test as well as the seven day larval test when the 

### 8 methods are finally validated or for use of that. We also 

### 9 support development protocols for chronic toxicity testing of 

### 10 both adult bees and the larvae; and the industries interested in 

### 11 participating in this process if it is possible. 

### 12 We also support the White Paper position that 

### 13 studies need to produce measured endpoints that can be clearly 

### 14 tied to assessment endpoints, such as colony strength and 

### 15 survival and the protection goals. We also feel that the 

### 16 internationally-common data requirements should have harmonized 

### 17 test guidelines so that we can use our studies for multiple 

### 18 countries and multiple regulatory agencies. 

### 19 For Tier II, as Dave mentioned also that we 

### 20 like to see flexible study designs to be able to be produced to 

### 21 address uncertainties in the lower tier assessments; and we also 

### 22 realize that it is critical to communicate with the agencies on 

### 23 the study design such that we design studies that help answer 

### 24 questions that are apparent or has been useful for the risk 

### 25 assessment purpose. 

### 26 I would also like to point out that within the

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1 semi-field studies, again we are subjecting the bees to a 

### 2 somewhat unrealistic exposure scenario, again force feeding on a 

### 3 plot of a crop where insecticide has been applied or pesticide 

### 4 has been applied. And that in these studies they can produce 

### 5 false negatives or fail to show effects that occur at real field 

### 6 sites; but often produce false positives that show effects that 

### 7 don't occur at real field sites. 

### 8 So again, this relates to the fact that we 

### 9 need to go to Tier III studies to help answer, again, some of 

### 10 these questions related to the uncertainty of Tier II. 

### 11 So again, as with the Tier II studies, we also 

### 12 feel that it is necessary to consult with the agencies to 

### 13 develop appropriate protocols and site designs to help answer 

### 14 the questions that are necessary for risk assessment purposes. 

### 15 Studies should be designed and endpoints selected to provide the 

### 16 most appropriate information to assess protection goals. 

### 17 Again, the study should be focused and help to 

### 18 answer specific questions rather than addressing all potential 

### 19 scenarios. Adding additional scenarios to study design creates 

### 20 more confusion as to how to interpret the results. So, keeping 

### 21 them focused would definitely be of benefit. 

### 22 The Tier III studies incorporate potential 

### 23 effects of the plant protection products. It is multiple levels 

### 24 of organizations, so we would incorporate information from lower 

### 25 levels of biological organization up through the population 

### 26 level. So, we feel like this is, you know, important that we

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1 focus on these higher level assessment endpoints that are able 

### 2 to relate to protection goals, such as protecting pollination 

### 3 services. 

### 4 I would also like to point out in field 

### 5 studies, control plots are really reference plots because in 

### 6 most situations we have to apply chemicals to the control plots 

### 7 in order to grow a viable crop for appropriate comparisons. And 

### 8 also I would like to indicate that real-world biological systems 

### 9 are inherently variable and the variability should be properly 

### 10 interpreted within these studies. 

### 11 The next two slides are a couple of graphs 

### 12 illustrating some data we obtained from some Tier III field 

### 13 studies. Again, these graphs are in the White Paper on page 

### 14 170. 

### 15 The first graph here is illustrating effects 

### 16 of individual mortality, and that would be dead worker bees, 

### 17 pupae and larvae over the course of the study. As we see here, 

### 18 at specific days throughout the course of the study, we see 

### 19 differences in mortality between treatments and controls. 

### 20 I would also like to point out that the 

### 21 highest level of mortality was observed prior to initiation of 

### 22 the treatment where the hives were brought into the actual study 

### 23 site. 

### 24 Throughout the course of the study we do see 

### 25 again, variability within individual mortality. If we move to 

### 26 or look more closely at higher level assessment endpoint such as

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1 colony strength -- and again this is estimated number of bees 

### 2 per hive -- we see that the controls and treatment track fairly 

### 3 well. 

### 4 So again, at this higher level endpoint, it is 

### 5 incorporating all of the lower level assessment endpoints, the 

### 6 lower levels of biological organization into this higher level 

### 7 of biological organization endpoint. We feel that this colony 

### 8 level type of assessment is where we need to be as far as 

### 9 determining effects of potential compounds on the health of 

### 10 honey bees. 

### 11 In reality, these assessment endpoints will 

### 12 then help looking toward protection goals. So, we are working 

### 13 towards determining endpoints that help to assess the protection 

### 14 goals as stated in the White Paper. With that, I will hand it 

### 15 back to Dave. 

### 16 **DR. DAVE FISCHER:**Okay. So, in terms of risk 

### 17 assessment, again we support a tiered approach for all the 

### 18 reasons we have said. It is consistent with the standard 

### 19 methodology the EPA uses for other taxa. 

### 20 Science-based approach to develop the proposed 

### 21 levels of concern, I mentioned that before; use of honey bees as 

### 22 surrogate for non-Apis bees; use of whole colony studies when 

### 23 triggered to put the results of laboratory testing with 

### 24 individual adult or larval bees into real-world context. This 

### 25 is an opportunity that we have with bees that we don't frankly 

### 26 have with some of the other species we do testing and risk

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1 assessment for. It is hard to do a population level test with 

### 2 birds, fish and mammals. 

### 3 Bees - that is what usually we want to protect 

### 4 is that level. With bees, we can test colonies and even acres. 

### 5 It is just an opportunity that we can test at that level that we 

### 6 don't always have. 

### 7 Communication of the results of pollinator 

### 8 risk assessments should convey the uncertainties and inherent 

### 9 conservatism of the assessments methods. That is something we 

### 10 think is key with these Tier I assessments that are going to - 

### 11 there are compounds that are going to not be screened out and 

### 12 the concern is that that is going to be communicated as there is 

### 13 a risk. It doesn't necessarily mean there is any risk. It 

### 14 means that with the conservatism in the assessment, you couldn't 

### 15 say there wasn't one. 

### 16 Population modeling - this is an area that a 

### 17 number of companies are investing in. We support the 

### 18 development of population modeling to address uncertainties in 

### 19 lower tier risk assessments. We are interested in participating 

### 20 in model development, and as I mentioned, we are investing some 

### 21 resources in that. Models should be transparent, have options 

### 22 for refinement and be well vetted before use by regulatory 

### 23 agencies. 

### 24 So, to summarize sort of our technical review 

### 25 of this, there are a number of points of agreement. We think 

### 26 the new methodology represents an important advancement that

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1 should improve the scientific credibility of pollinator 

### 2 assessments and increase the transparency and predictability of 

### 3 regulatory decisions. That's a good thing. We support the 

### 4 efforts of EPA, PMRA and CDPR to harmonize their pollinator risk 

### 5 assessments processes. Pollinator assessments should be based 

### 6 on scientifically sound methods and we think the proposal is 

### 7 consistent with that goal. Risk assessment should be 

### 8 interpreted within the risk-benefit context as required by 

### 9 FIFRA. That is particularly well explained in the White Paper. 

### 10 11 There are some areas where we would suggest 

### 12 there are opportunities for improvement. Proposed screening 

### 13 criteria appear in some cases to be overly conservative and will 

### 14 produce many false positives. Many chemicals that pose no 

### 15 real-work risk to pollinators will fail the Tier I screen. 

### 16 There was some information presented on that, but I just thought 

### 17 I would throw up here that if you apply the Tier I dietary risk, 

### 18 it's basically - there is a fixed assumed dose per 1 pound per 

### 19 acre applied that is something like 32 micrograms that the bee 

### 20 ingests. 

### 21 So, how high does your - I'm sorry I have to 

### 22 go back to my spread sheet - but basically to pass the 

### 23 assessment, you need an LD50 - if you have 1 pound per acre, you 

### 24 need to have an LD50 of 80 if you are applying 5 pounds per 

### 25 acre, the LD50 has to be above 400. We only test up to 100, so 

### 26 any chemical that is slightly above 1 pound per acre is not

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1 going to pass this assessment, whether it is toxic or not unless 

### 2 we test higher. We think that the limit test should be enough. 

### 3 If you are down at 1/10 of a pound a.i. per acre, you need to 

### 4 have an LD50 higher than 8, which again that is a very low rate 

### 5 and that is not a particularly low LD50 number. 

### 6 Just comparing it to the Hazard Quotient 

### 7 method in Europe - in Europe there is sort of a 

### 8 back-of-the-envelope calculation, which the application rate is 

### 9 compared to the LD50 and you compare that result, that Hazard 

### 10 Quotient to a trigger value of 50 and to pass that to get a 

### 11 Hazard Quotient that is less than 50 for 5 pound-per-acre 

### 12 application rate, the numbers are up there - 112 micrograms per 

### 13 bee is what the LD50 has to be for 1 pound per acre, 22, all of 

### 14 these are 3.6X different. 

### 15 So this North American approach is 3.6X more 

### 16 conservative than the EU Hazard Quotient approach; and it's all 

### 17 because of the assumed dose that these are getting from nectar 

### 18 and the fact that you are estimating the nectar concentrations 

### 19 to be as high as they are. 

### 20 And by the way I think everybody in Europe is 

### 21 pretty comfortable that Hazard Quotient is very conservative. 

### 22 We are going even more conservative with this one. 

### 23 I mentioned that if you have a chemical that 

### 24 is nontoxic at the limit dose in the acute oral test then these 

### 25 chemicals should just be considered safe and you should not 

### 26 proceed to higher tiers.

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1 It is important the Tier I assessments are 

### 2 used only to identify chemicals requiring further testing and 

### 3 not for concluding or implying that such chemicals pose a 

### 4 real-world risk, I mentioned that. 

### 5 Since higher tier testing will be triggered 

### 6 for many chemicals, there must be clear guidance and achievable 

### 7 acceptability criteria for higher tier studies. I mentioned 

### 8 that at the outset. I will mention it again. That is a 

### 9 concern. We have a long history of doing higher tier tests on 

### 10 other taxa, for which there is a high rejection rate of the 

### 11 studies when they are reviewed by the Agency. So, this is an 

### 12 area where we see the need for a lot of discussion, maybe 

### 13 workgroup, research, whatever and we are interested in 

### 14 participating in the process. 

### 15 Just as a last slide, industry continues to be 

### 16 proactive in advancing the science related to bee health issues 

### 17 such as conducting research programs, developing best management 

### 18 practices, formulation improvements, stewardship programs, 

### 19 pollinator habitat initiatives, and this is an area that we are 

### 20 active in and we want to continue to collaborate with the 

### 21 agencies and with the research community to continue to advance 

### 22 the science. Thank you for your attention. 

### 23 **DR. DANIEL SCHLENK:**Thank you for your 

### 24 comments. Questions from the panel? Dr. Hunt? Dr. Hunt is 

### 25 going to go first. 

### 26 **DR. GREG HUNT:**I wonder if given the problems

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1 with Tier III testing and getting an acceptable study that EPA 

### 2 would accept, would CropLife be open to the idea if EPA only 

### 3 required Tier II testing? 

### 4 **DR. DAVE FISCHER:**Jay might want to answer 

### 5 this as well, but I think we definitely want to have the option 

### 6 for field testing. At Tier II, we exaggerate the exposure when 

### 7 we do bee tunnel tests. We know of cases where there are 

### 8 affects in those tests that are basically false positives with 

### 9 respect to what happens in the field because the exposure is 

### 10 exaggerated. There is also the problem - I think the Agency 

### 11 gave a good summary of all the deficiencies or short comings of 

### 12 tunnel studies in that if you stress the bees you have limited 

### 13 amount of time that you can keep them. 

### 14 So, I think we want to definitely have the 

### 15 option of doing full field testing and the challenge is to 

### 16 develop protocols where the results are clearly interpretable 

### 17 and everybody agrees on how to interpret the results. 

### 18 **DR. GREG HUNT:**On the other hand, full field 

### 19 testings may lead to false negatives. I just wanted to make 

### 20 that comment. 

### 21 **DR. DANIEL SCHLENK:**Thanks. Dr. Overmyer, 

### 22 did you want to add anything to that? 

### 23 **DR. JAY OVERMYER:**No. I agree with what Dave 

### 24 had mentioned. 

### 25 **DR. DANIEL SCHLENK:**Dr. James, you said you 

### 26 had a - no? Okay. Anyone else? Dr. Potter?

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1 **DR. THOMAS POTTER:**With regard to Tier II and 

### 2 Tier III, where does evaluation of mitigation practices fit into 

### 3 the framework and the approach? 

### 4 **DR. DAVE FISCHER:**We see mitigation as being 

### 5 an option at every stage of when we develop a product. Before 

### 6 we even submit it for a registration, we are determining what 

### 7 the hazards are to various taxa and developing a label with 

### 8 potential mitigation in mind; so some products, people like 

### 9 myself and Jay are asked by the company, do we need an at bloom 

### 10 restriction? Do we need to off label this crop or that crop 

### 11 because of particular attractiveness to pollinators. 

### 12 So, that is part of preparing the label and as 

### 13 you work through the system there should be an option for - 

### 14 instead of conducting another million dollars of work, 

### 15 mitigating the problem with a label restriction. 

### 16 **DR. DANIEL SCHLENK:**Yes, Dr. Pettis? 

### 17 **DR. JEFF PETTIS:**Yeah, Jeff Pettis. With 

### 18 regard to systemics and trying to predict concentration in 

### 19 nectar and pollen, do you feel that what is found in stems or in 

### 20 the plant tissue can accurately predict that? And also could 

### 21 you comment on just the ratio which we find in the few studies 

### 22 that we have about the concentrations in pollen versus nectar? 

### 23 **DR. JAY OVERMYER:**From the data that we 

### 24 generated, we typically see higher levels in the actual plant 

### 25 leaves and stems compared to what we see in nectar and pollen, 

### 26 but we do see variability within different plants. So, I think

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1 it is important that we indicate that in generalization you 

### 2 cannot take one example and then extrapolate it to all different 

### 3 examples. As far as nectar and pollen goes, we typically do see 

### 4 more residues of pollen than nectar. Again, the ratios there 

### 5 vary between different plants. 

### 6 **DR. DAVE FISCHER:**This is Dave Fischer. I 

### 7 will just add to that that it may differ between the chemistry, 

### 8 the compound involved. There are definitely a lot of chemicals. 

### 9 Most of the systemic chemicals are largely xylem-limited in 

### 10 their transport in the plant. They will move with the water 

### 11 transport system, but not with the sugar transport system, but 

### 12 then there are some other chemicals that have the right chemical 

### 13 properties that will also be systemic in the phloem. 

### 14 So, just looking at data and saying what is 

### 15 the concentration of stem versus the flowers, you really 

### 16 probably need to look a little bit more carefully about, you 

### 17 know how does that compare based upon the chemical properties 

### 18 that would affect their ability to move in these different 

### 19 transport systems. But as a general rule, the stem and leaf 

### 20 concentrations are an order of magnitude higher than what is in 

### 21 the flowers - what gets in the pollen and nectar. 

### 22 **DR. DANIEL SCHLENK:**Okay. Any other 

### 23 questions? Okay. Thank you very much. Our next public 

### 24 commenter is CropLife Canada on my list. Presenter is Pierre 

### 25 Petelle. 

### 26 **MR. PIERRE PETELLE:**Good morning. So, yeah,

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1 my name is Pierre Petelle. I am the Vice President of Crop 

### 2 Protection Chemistry at CropLife Canada. I just have a few 

### 3 slides to do through, and you will see probably a lot of 

### 4 repetition from what we just talked about because we do 

### 5 collaborate quite a bit with our colleagues to the south here. 

### 6 Our technical working group in Canada, we 

### 7 actually share some members with the technical team here at CLA. 

### 8 We collaborate on this file in particular because it is a very 

### 9 important file for the industry. CropLife Canada and our 

### 10 members support the CLA position and the comments you just heard 

### 11 on the White Paper. 

### 12 As indicated previously, pollinators and 

### 13 pesticides are complementary components of the sustainable 

### 14 agricultural system. Many of the crops grown in Canada, like 

### 15 here in the U.S., depend on both crop protection and pollination 

### 16 services. 

### 17 So, a sound scientific approach to 

### 18 understanding impact of pesticides on pollinators is going to be 

### 19 absolutely critical here. 

### 20 As Ray pointed out earlier, bee health is a 

### 21 complex issue. Our industry is committed to the protection and 

### 22 promotion of pollinator health, and we are very supportive of 

### 23 efforts to advance the science. So, as we move forward in this 

### 24 process, we want to reiterate the need for scientifically sound 

### 25 risk assessment procedures and the use of data from 

### 26 well-designed, valid scientific studies.

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1 Canada and the U.S. enjoy a long history of 

### 2 cooperation and collaboration in the pesticide regulatory area. 

### 3 PMRA and EPA in particular have been world leaders in the area 

### 4 of regulatory harmonization, pesticide reviews, undertaking 

### 5 joint reviews, work sharing projects and just improving the 

### 6 scientific processes. I think this has helped both growers and 

### 7 regulators and our industry on both sides of the border in terms 

### 8 of access to newer safer products, in terms of using the most 

### 9 modern science and in terms of efficient regulatory review. 

### 10 This process has been developing over the past 15 years at 

### 11 least, and you know, I think as we move forward with this 

### 12 particular issue, we are highly supportive of harmonized data 

### 13 requirements, test guidelines and risk assessment processes. We 

### 14 are glad to see the role that PMRA has played in this whole 

### 15 process with the White Paper and here at this Panel. 

### 16 So just lastly, this is a reiteration of some 

### 17 of the comments that CLA made a few minutes ago, but in order to 

### 18 ensure the protection goals are met and instilling public 

### 19 confidence, we circle back to this need for a scientifically 

### 20 sound approach to risk assessment. We feel that the proposed 

### 21 approach is broadly consistent with this goal looking forward. 

### 22 In particular, we want to stress the 

### 23 importance of the tiered risk assessment approach that was just 

### 24 discussed in detail, and the use of honey bees as a good 

### 25 surrogate for non-Apis. 

### 26 That's all I wanted to say today. Thanks very

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1 much, and we again appreciate the role of PMRA in this process 

### 2 and stress the importance of scientifically rigorous process as 

### 3 we move forward. 

### 4 **DR. DANIEL SCHLENK:**Any questions from the 

### 5 panel? Dr. Berenbaum? 

### 6 **DR. MAY BERENBAUM:**Given the differences that 

### 7 exist in apicultural practices in Canada and the U.S. and 

### 8 variance on reconstraints, do you think standards are completely 

### 9 harmonizable, or do you think that there still may be means for 

### 10 recognition of differences? 

### 11 **MR. PIERRE PETELLE:**Yeah, I mean I think in 

### 12 terms of the science and the risk assessment process, I think 

### 13 there is an ability to harmonize those processes. Are there 

### 14 Canadian specific or U.S. specific practices that need to play 

### 15 into that? I mean that is going to have to be a question as we 

### 16 move forward, but from our experience up to this date, 

### 17 harmonization of those processes should be fine. 

### 18 **DR. DANIEL SCHLENK:**Thank you very much. 

### 19 **MR. PIERRE PETELLE:**Thank you. 

### 20 **DR. DANIEL SCHLENK:**Our next presenter is the 

### 21 Center for Regulatory Effectiveness, and that is Mr. William 

### 22 Kelly. 

### 23 **MR. WILLIAM KELLY:**I do not have any slides 

### 24 or handouts. My name is Bill Kelly. I am with the Center for 

### 25 Regulatory Effectiveness. We are an independent organization 

### 26 that evaluates regulatory issues that are of widespread concern

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1 and usually have quite a bit of controversy. We are also 

### 2 placing great emphasis on what we call information quality and 

### 3 the Information Quality Act and its guidelines. 

### 4 We have already submitted some detailed 

### 5 written comments, lots of footnotes and quotations, so I won't 

### 6 bother you with that. 

### 7 Dr. Steeger yesterday at the end of the 

### 8 meeting emphasized that the high quality of EPA studies and the 

### 9 studies that are conducted or required to be submitted under 

### 10 their guidelines, and we recognize that. I think that the 

### 11 problem we have is with a lot of journal articles. 

### 12 The Information Quality Act Guidelines 

### 13 recognize that peer review of journal articles is of very uneven 

### 14 quality. A lot of articles are published simply because they 

### 15 are, let's say, thought provoking and not really contributory to 

### 16 the knowledge base. Those need to be examined carefully for 

### 17 compliance with the standards and guidelines under the IQA. 

### 18 Near the end of yesterday's meeting, the Panel 

### 19 asked a good question that relates to the Information Quality, 

### 20 and that is what to do with data gaps. 

### 21 Data gaps are not really a problem with Tier I 

### 22 or Tier II, but when you get to the final risk assessment and 

### 23 you have gone through Tier III, the way the data gaps have to be 

### 24 handled is a matter of law under the IQA and its guidelines is 

### 25 for the scientists -- that are the risk assessors as opposed to 

### 26 the risk managers -- to simply describe the data gap and its

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1 significance, and then leave it to the risk managers to decided 

### 2 what to do with that. The data gap should not be filled with 

### 3 assumptions or default values or whatever. Those are usually 

### 4 policy-driven values that should not be used in the risk 

### 5 assessment process. 

### 6 Another good example that came up very late 

### 7 yesterday after unfortunately a lot of people had left the 

### 8 meeting was the issue of what we are really dealing with here. 

### 9 The extent and I would say also the distribution of the 

### 10 pollinator decline, which is often equated with colony collapse 

### 11 disorder unfortunately. A chart was presented, which showed the 

### 12 decline was presented by Mr. Steiger or Mr. Carrington. Then we 

### 13 found out that that chart was based on honey production. So, 

### 14 that would not pass muster under the Information Quality Act 

### 15 Guidelines for example. 

### 16 I mean we really do need good data on the 

### 17 extent of this problem and its distribution. I think that that 

### 18 is important, especially for Tier I, especially the 

### 19 distributional aspect of it. I mean if you look at distribution 

### 20 of the problem and the extent of the problem in various areas, 

### 21 you might find out that there are say, crop areas. We're not 

### 22 talking about states necessarily, but crop areas or particular 

### 23 crops where there is a high level of decline, but a particular 

### 24 pesticide is not used on those crops or in that area. That 

### 25 could be a basis for eliminating that pesticide from Tier I. 

### 26 On the other hand, you could have an area in

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1 which a particular pesticide is used very intensively, and yet 

### 2 there is no significant increase in pollinator decline beyond 

### 3 what you would expect from, you know, say ordinary winter kill. 

### 4 So again, that could factor into Tier I determinations. 

### 5 This issue of the extent of pollinator decline 

### 6 and the distribution of the problem is really missing from the 

### 7 framework at this point. There could be an interagency 

### 8 coordination issue here because it appears that USDA has taken 

### 9 the lead on this issue. 

### 10 Back in January, the Inspector General of the 

### 11 Department of Agriculture issued a report that was on progress 

### 12 the USDA had made in evaluating this issue. The quote from the 

### 13 report says, "The true extent and impact of CCD -- which they 

### 14 equated with general decline -- in the United States has not 

### 15 been adequately assessed despite USDA's use of significant 

### 16 resources for honey bee research to address CCD." 

### 17 The EPA is part of that steering committee 

### 18 that is working on this issue. And as I understand it, part of 

### 19 the problem is money either from Congress or with redirecting 

### 20 resources within the agencies. 

### 21 So, I think in looking at the matter of 

### 22 problem formulation, this issue is really missing from the 

### 23 framework at this point and needs to be added. The SAP needs to 

### 24 address it. We need to do some sort of pattern analysis on 

### 25 where this problem is occurring, the extent of it and whether it 

### 26 is really associated with pesticide use in those ares.

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1 And finally, I think if we are going to do 

### 2 this since it is missing from the framework at this point, we 

### 3 need some further public comment on this particular aspect of 

### 4 the issue. Okay. That is all I have at this point. Are there 

### 5 any questions? 

### 6 **DR. DANIEL SCHLENK:**Thanks. Dr. Berenbaum? 

### 7 **DR. MAY BERENBAUM:**Actually, I am a little 

### 8 confused. In view of the fact that bees are livestock and 

### 9 there's at least some legal precedence that bees are owed the 

### 10 duty of common humanity if they trespass on agricultural land, 

### 11 why is documenting decline relevant to risk assessment of 

### 12 pesticide use on agricultural land? 

### 13 **MR. WILLIAM KELLY:**Thank you. Well, 

### 14 presumably this framework and this risk assessment could lead to 

### 15 FIFRA Regulatory Action, particularly on registration. FIFRA, 

### 16 as a matter of law, requires a balancing of risks and benefits. 

### 17 In order to do that, you are going to need to quantify the risks 

### 18 together with the benefits, and you can't do that with some 

### 19 accurate appreciation of the extent of the impact of what's 

### 20 occurring. Does that. 

### 21 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 22 **DR. MAY BERENBAUM:**Well as you pointed out, 

### 23 bee decline is not equal to bee mortality. So, bee death per se 

### 24 isn't colony collapse disorder, but still risk assessment for 

### 25 pesticide use involves determining bee susceptibility, bee 

### 26 mortality. So, I am still not sure where the linkage to

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1 documenting decline, at least for honey bees. 

### 2 **MR. WILLIAM KELLY:**I think what we are 

### 3 interested in is decline. I don't know how we would measure 

### 4 decline in health short of colony loss and bee mortality. At 

### 5 least that's how the problem has been posed to date in terms of 

### 6 colony loss and mortality. If we want to start looking for sick 

### 7 bees, I guess that takes us to another level. 

### 8 **DR. DANIEL SCHLENK:**Okay. Any other 

### 9 questions or comments? Okay. Thank you very much. Our next 

### 10 presenter is Scott Schertz with the National Agricultural 

### 11 Aviation Association. 

### 12 **MR. SCOTT SCHERTZ:**Hello. I do appreciate 

### 13 the opportunity to be in front of the SAP on the pollinator 

### 14 protection issues, and we do support this initiative. 

### 15 Basically, well my name is Scott Schertz. I 

### 16 do own and operate Schertz Aerial Service, an aerial application 

### 17 operation in central Illinois in corn-soy-bean country. I am a 

### 18 past president of the NAAA and the comments have to do with 

### 19 basically aerial application of crop protection products. 

### 20 The NAAA does represent a majority of the 

### 21 aerial application industry in the U.S. There are approximately 

### 22 1700 members in 46 states, and the majority of the members of 

### 23 the NAAA are operations such as mine or small or medium-sized 

### 24 businesses that provide these services to many crops, everywhere 

### 25 from the major crops to many of the specialty crops. 

### 26 As the White Paper reflects that mode of

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1 application is one of the areas of concern, you know I would 

### 2 like to give a little introduction to the aerial application 

### 3 industry as far as our scope and qualifications. 

### 4 Aerial application does account for 

### 5 approximately 25 percent of the application of crop protection 

### 6 products in the U.S. and commercial bases, and in many areas, it 

### 7 is the preferred ways of applying due to its speed, able to get 

### 8 over tall crops or in conditions that make ground application 

### 9 impractical. 

### 10 In addition, that means that aerial 

### 11 application does have, you know, a unique opportunity on 

### 12 pollination protection in that the airplanes and helicopters can 

### 13 apply a lot of product very rapidly and in a narrow time window. 

### 14 When the hives - the areas of the prime bee activity are known 

### 15 as many states required, this does allow a tiered approach, 

### 16 basically applying at time of low pollinator activity. 

### 17 I do want to emphasize though, that that 

### 18 doesn't mean that it is practical for all applications to be 

### 19 done then; and that knowledge of where they are is very, very 

### 20 important as I will emphasize many states require. 

### 21 When you start looking at the crop protection 

### 22 needs of, you know, many of the major field crops that I am very 

### 23 familiar with, there is a need for viable products with 

### 24 reasonable label requirements, even during pollination time. I 

### 25 mean, that is when the majority of the insects actually affect 

### 26 plants, and obviously there is a conflict here in times.

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1 And then also our members do serve many of the 

### 2 specialty crops that may be intensively utilizing managed 

### 3 pollinators, and in those cases, I mean these types of things of 

### 4 spraying at night are common. But it doesn't, I mean, it isn't 

### 5 a matter of just getting more airplanes. I mean, this is a very 

### 6 specialized industry. There are a finite number of airplanes. 

### 7 The airplanes are very sophisticated. There is a lot of 

### 8 technology in them that I will go through briefly in a couple 

### 9 minutes. 

### 10 Then also the night operation is a very 

### 11 specialized subpart of it. Yet, some areas it is common; 

### 12 typically it is in more of the specialty crop areas. In my 

### 13 area, yes I do some night work, but there are also many fields 

### 14 that it is impractical and unsafe to do at night. When you 

### 15 start talking about dealing with obstacles such as met towers, 

### 16 winter events, large power lines, the judgement at night, the 

### 17 depth perception is a real issue. So, you can't just say well 

### 18 that is going to magically fix all of these issues. 

### 19 The industry is very active on refining the 

### 20 safety programs and the deposition of the airplanes. Our main 

### 21 program is something called PASS (phonetic) that has been in 

### 22 effect for about 14 years. It is something that is an active 

### 23 training session that goes around the country and is available 

### 24 to the entire industry. Basically, the subjects have to do with 

### 25 drift mitigation, aeronautical decision making, security and 

### 26 pollinator issues have been something that has been addressed in

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1 the last several years. 

### 2 Another unique thing about the aerial 

### 3 application industry is what is called our safe program. 

### 4 Obviously, these details are in the paper submitted, and that is 

### 5 a program that is also unique to aerial application industry on 

### 6 an infield way of measuring the deposition from the airplanes 

### 7 and refining that to be appropriate for the products to be used. 

### 8 9 So, I did want to give you a bit of 

### 10 introduction and how that, you know, aerial application does 

### 11 have some unique opportunities on this, but you will also need 

### 12 to be realistic that it isn't a silver bullet of just doing 

### 13 everything at night. 

### 14 There is further detail in the paper that was 

### 15 submitted that you should have a copy of. I do appreciate that 

### 16 opportunity to be in front of you. Any questions? 

### 17 **DR. DANIEL SCHLENK:**Any questions from the 

### 18 Panel? Okay. Thank you Mr. Schertz. 

### 19 **MR. SCOTT SCHERTZ:**Thank you. 

### 20 **DR. DANIEL SCHLENK:**Our next presenter is 

### 21 James Doan, who is an owner of a bee keeping operation. 

### 22 **UNIDENTIFIED SPEAKER:**Mr. Doan is running 

### 23 late. 

### 24 **DR. DANIEL SCHLENK:**Okay. Is Mr. Jenkins 

### 25 available, for Center for Food Safety? 

### 26 **MR. PETER JENKINS:**Greetings. I am Peter

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1 Jenkins, Attorney and Consultant working with the Center for 

### 2 Food Safety. You should have copies of the oral comments that I 

### 3 am going to give you now. 

### 4 We are in contact with Mr. Doan, and he is 

### 5 hung up on the Metro somewhere. Hopefully he will be here 

### 6 before the end of the comment period. Dr. Jenkins if you could 

### 7 be a bit lenient on that, and we will let you know when he shows 

### 8 up. He is a beekeeper who came from New York. 

### 9 Let's see, I am just going to basically read 

### 10 my comments. We have written comments that are in the record 

### 11 and I will be glad to answer any questions at the end about 

### 12 either the oral or written comments. 

### 13 I am pleased to make these comments on the 

### 14 White Paper that was submitted in support of the risk assessment 

### 15 process or the proposed RA process for short. 

### 16 My organization, the Center for Food Safety, 

### 17 has got comments in your packet. Please give those your close 

### 18 attention because our comments as well as the comments of Beyond 

### 19 Pesticides and some of the other NGOs involved here -- and 

### 20 beekeepers -- have submitted some detailed scientific studies 

### 21 that are omitted from the proposed RA process document that we 

### 22 think very much relate to this process. 

### 23 Let me stress the critical role you are 

### 24 playing at ensuring a state-of-the art risk assessment framework 

### 25 to protect honey bees and native bees from excessive and perhaps 

### 26 unsustainable mortality from pesticides. It appears,

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1 unfortunately, that EPA lacks inhouse PhD levels entomological 

### 2 expertise, at least very much of it that we can see. So, you 

### 3 can help to provide that. 

### 4 The process you are a part of has involved 

### 5 real world cost and impacts already. Years of delay and 

### 6 thousands of staff hours have gone into creating the proposed 

### 7 framework, but it still has major areas of uncertainty and 

### 8 omission. Further, EPA has refused to follow through on the 

### 9 prior condition that had it required years ago when it 

### 10 registered two neonicotinoid insecticides, Clothianidin and 

### 11 Thiamethoxam, for which providing a valid bee impact field 

### 12 study, a pollinator field study or Tier III sort of test as we 

### 13 have been hearing about, was made a condition to continue in the 

### 14 registration of those pesticides by EPA. That condition has 

### 15 still not been met nine years after it was first imposed in the 

### 16 case of Clothianidin. 

### 17 An excuse that EPA now uses is its condition 

### 18 currently is being "held in reserve" pending the outcome of this 

### 19 FIFRA SAP process. So you, as SAP Panelists, have become part 

### 20 of EPA's non-precautionary approach to this issue of pollinator 

### 21 field test design for these pending conditional registrations. 

### 22 So, now you can at least help make sure that 

### 23 the cross bar on the shifted goal post is high enough to be 

### 24 really comprehensively protective. It has taken so long to get 

### 25 to this point. 

### 26 A written comment highlighted the inadequacy

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1 of the proposed RA framework as far as taking sublethal effects 

### 2 into account. Our comment also urged modeling for both the 

### 3 contaminated dust and the guttation fluid exposure routes for 

### 4 systemic neonicotinoid insecticides, and to have a clear call 

### 5 out in the RA framework for more Tier II and Tier III testing, 

### 6 if necessary, of those roots before approving such insecticides 

### 7 in the future. 

### 8 On the neonicotinoid contaminated dust route, 

### 9 it was very inappropriate for the RA framework on page 99 to 

### 10 include evasive statements from the agency that in effect say, 

### 11 the SAP can just rely on EPA to mitigate the risks of this route 

### 12 through discussions with seed treatment registrants and the 

### 13 manufacturers of seed planting equipment. 

### 14 That sort of vague, hopeful, self-serving 

### 15 assertion restated at least twice by EPA officials yesterday has 

### 16 no place in a risk assessment framework. 

### 17 EPA has no real power to enforce such a 

### 18 speculative, undefined requirement for modifying tens of 

### 19 thousands of planting machines across the entire country, and 

### 20 even if EPA had such power, the practical obstacles to EPA 

### 21 enforcing that sort of requirement in any reasonable timeframe 

### 22 far exceed the Agency capabilities. 

### 23 This contaminated dust exposure route and its 

### 24 impact on honey bees has led to Germany, Italy, Slovenia, to 

### 25 some extent France, prohibiting various neonicotinoid seed 

### 26 treatments in the last four years. So relying in EPA's attempts

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1 to downplay this route in the U.S. would be irresponsible. 

### 2 The SAP must demand a more rigorous treatment 

### 3 of this route or it won't be doing its job. Please take careful 

### 4 account on this issue of the written comments by Dr. Christian 

### 5 Krupke of Purdue University that in the packets - I hope they 

### 6 are in your packets - which reinforce what I have just said. 

### 7 Proposed RA framework document makes 

### 8 occasional mention of EPA's ecological incident information 

### 9 system or EIIS. I would remind the SAP members that it is well 

### 10 accepted across numerous stake holders, especially beekeepers, 

### 11 that the EIIS does a very poor job of collecting national bee 

### 12 kill incident data. 

### 13 In contrast, Canada's PRMA reporting system is 

### 14 well regarded. Here is an illustration. In EPA's July 27, 2012 

### 15 response letter to a petition filed by my organization, the 

### 16 Center for Food Safety, 25 beekeepers and other NGOs to suspend 

### 17 the registration of Clothianidin, the Agency stated with respect 

### 18 with EIIS, "We are however aware of 14 additional incidents 

### 19 occurring in the U.S. in 2012 that are not yet present in the 

### 20 database and approximately 120 additional incidents reported in 

### 21 Canada." 

### 22 Extensive media and non-EIIS reports make 

### 23 clear that both Canadian and U.S. beekeepers suffered vast 

### 24 numbers of spring 2012 bee kills due to the contaminated dust 

### 25 exposure route associated with corn planting just this past 

### 26 spring. It is not plausible at all that 120 documented bee kill

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1 incidents associated with neonicotinoid seed treatments 

### 2 involving several thousand bee colonies occurred in Canada's 

### 3 relatively small corn planting area -- mostly in southwestern 

### 4 Ontario and Quebec as we understand it -- but only 14 additional 

### 5 bee kill incidents occurred in the entire U.S. across its orders 

### 6 of magnitude larger corn areas. 

### 7 The seed treatment products, machinery used 

### 8 and environmental conditions do not change dramatically when you 

### 9 cross the border. What changes is the reliability of the bee 

### 10 kill reporting systems. My point here is that a revised RA 

### 11 framework should not rely on the EPA, EIIS unless it is 

### 12 dramatically improved. 

### 13 On the issue of drinking water - our written 

### 14 comments focused on the guttation liquid route and those are 

### 15 important comments, but I urge your attention to a separate 

### 16 exposure route that our written comments did not really address. 

### 17 A key paper by Starner and Goh 2012, is Detections of the 

### 18 Neonicotinoid Insecticide, Imidacloprid, in Surface Waters of 

### 19 Three Agricultural Regions in California. It documents that a 

### 20 significant portion of sampled surface waters were contaminated 

### 21 above EPA allowed levels for chronic invertebrate exposure 

### 22 across diverse agricultural landscapes. 

### 23 These waters are wide-spread sources of 

### 24 drinking water for all bees, but the proposed RA framework does 

### 25 not mention this California DPR paper, nor does it quantify the 

### 26 risk to bees from the real world levels that were measured in

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1 that study. Instead, it follows a very questionable academic 

### 2 exercise in appendix 2 in the RA framework to discount this 

### 3 drinking water route all together. 

### 4 The fact this environmental contamination by a 

### 5 major neonicotinoid exists now in California and elsewhere is 

### 6 indicative of Agency failure to present undue consequences and 

### 7 it has passed inadequate risk assessments. The SAP can now 

### 8 correct this failing going forward. 

### 9 On the portion of the document addressing the 

### 10 risks to non-Apis bees, I think any fair appraisal of the 

### 11 framework document would conclude that this is its weakest 

### 12 section. After years of delay, merely suggesting that there 

### 13 might be future "modifications" of the basic RA framework to 

### 14 address the unique circumstances of thousands of native North 

### 15 American bee species is utterly inadequate to the risks that you 

### 16 are facing here. 

### 17 Given that many native species have small 

### 18 localized ranges, the RA process must at least explicitly assess 

### 19 the need to restrict or limit the use of pesticides in these 

### 20 limited locations, which is a common sense way to reduce risk to 

### 21 these localized species. Several of these non-robust species 

### 22 already are disappearing and lack any margin for Agency error 

### 23 moving forward and ensuring their survival. 

### 24 To the extent that the proposed RA framework 

### 25 has overlooked, down played or minimize various exposure routes, 

### 26 the classes of sublethal effects, the existing U.S. National Bee

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1 Kill Baseline and the other issues such as cumulative and 

### 2 synergistic impacts that I can't go into, the document 

### 3 constitutes a best case scenario from the perspective of the 

### 4 companies that you have heard from seeking to approve 

### 5 pesticides. But it is a worst-case scenario from the 

### 6 perspective of bee conservation and preservation and preserving 

### 7 beekeeper livelihoods. 

### 8 You hold the intellectual key to finding the 

### 9 right medium between these best and worst case scenarios. If 

### 10 you fumble an important element of the risk framework, you could 

### 11 put the long-term sustainability of these species, particularly 

### 12 the thousands of relatively poorly studied native bee species in 

### 13 jeopardy. 

### 14 I want to hit one last point because it just 

### 15 came up in a previous talk by CropLife. I have been looking at 

### 16 the RA framework on page 170. There are two figures, figures 12 

### 17 and 13; and some generalizable point is made about those 

### 18 figures. It is in our written comment. We said what are the 

### 19 labels, what is the citation, what is going on in these figures 

### 20 12 and 13, because they are basically cleansed of any 

### 21 information except to try to make a generalized point. 

### 22 Well, it turns out that those figures are the 

### 23 same ones that the gentleman from Syngenta showed in his slides. 

### 24 He actually had labels on his slides indicating what was going 

### 25 on there. 

### 26 So, it appears that EPA has just taken

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1 Syngenta's figures, put them into the RA framework at page 170 

### 2 without the labels and tried to make a generalizable point about 

### 3 it, but the point doesn't flow and it doesn't make sense. So, I 

### 4 just bring that to your attention. It's a bit concerning that 

### 5 EPA has used an unattributed bit of information from Syngenta or 

### 6 CropLife, whoever it was that put it together and just stuck it 

### 7 into the document. 

### 8 So, with that, again I am an attorney. I 

### 9 cannot answer scientific questions, but I am glad to answer any 

### 10 questions that I can. Thank you. 

### 11 **DR. DANIEL SCHLENK:**Thank you for your 

### 12 comments. Any questions from the Panel? Okay. So let's go 

### 13 ahead and take a break right now - 15 minutes - be back at 

### 14 10:45. We will continue with our public comments at that time. 

### 15 16 (Brief recess.) 

### 17 Okay. If I can have everybody take their 

### 18 seats please. Okay. Is James Doan here? Yes. Okay. Mr. 

### 19 Doan, if you don't mind, we are going to have you come second 

### 20 after Beyond Pesticides comes forward if that's okay. Sorry 

### 21 about that. 

### 22 So, can I have Nichelle Harriott, is she in 

### 23 the room? As Nichelle is coming forward, just to point out, I 

### 24 believe Dr. Fefferman you had asked for the OECD Guidance 

### 25 Document on the Validation of Structure Activity Relationship 

### 26 Models? That is on the docket. So, for anybody else who would

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1 like to do that, just go onto the website and you can down load 

### 2 a copy of that. Okay. Ms. Harriott? 

### 3 **MS. NICHELLE HARRIOTT:**Hi, good morning. 

### 4 Thank you for allowing me to present these comments. My name is 

### 5 Nichelle Harriott. I am representing Beyond Pesticides. Beyond 

### 6 Pesticides is a not-for-profit organization that works at the 

### 7 grassroots level to protect public and environmental health, 

### 8 identifying the hazards of chemical-intensive land, building and 

### 9 community management practices, and promoting healthy, 

### 10 sustainable and organic systems. 

### 11 Beyond Pesticides is very concerned about the 

### 12 status of our nation's pollinators, especially the honey bee. 

### 13 Each year since 2006, commercial beekeepers have reported 

### 14 unprecedented bee losses. Normal losses occur around 17.6 

### 15 percent, but over the 2008 and 2009 period an estimated 29 

### 16 percent of all U.S. bee colonies died. 

### 17 Since the honey bee is the most economically 

### 18 valuable pollinator worldwide, responsible for the pollination 

### 19 of high value crops such as almonds and broccoli, it is 

### 20 important that these pollinators are protected from the 

### 21 potential harmful effects of pesticides under EPA's 

### 22 jurisdiction. 

### 23 One class of pesticides in particular has 

### 24 emerged as a major suspect in recent bee decline. Several 

### 25 studies have demonstrate that neonicotinoid insecticides like 

### 26 Imidacloprid, Clothianidin and Thiamethoxam have sublethal

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1 effects in honey bees, which include disruptions in mobility, 

### 2 navigation and feeding behavior. Lethal and sublethal exposures 

### 3 have been shown to decrease foraging activity along with 

### 4 olfactory learning performance and decreased hive activity. 

### 5 When exposed to sublethal doses of 

### 6 neonicotinoids at levels present in the environment, honey bees 

### 7 were less likely to return to the hive after foraging. Bees 

### 8 exposed to these chemicals also exhibit convulsions, 

### 9 uncoordinated movements and tremors that are typical of 

### 10 neurotoxic insecticide exposure. One study analysis of dead or 

### 11 dying bees by Krupke et al. found residues present at the range 

### 12 of 3.8 to 13.3 parts per billion. 

### 13 A study by Schneider et al. shows that under 

### 14 field-like conditions, as little as 0.05 to 2 nanogram per bee, 

### 15 Clothianidin can significantly reduce foraging activity. In 

### 16 context, Reetz et al. found that corn seeds treated with 

### 17 Clothianidin result in translocated concentrations up to 8000 

### 18 nanograms per milliliter. For the honey bee, toxicity has been 

### 19 observed at an LD50 of 22 nanograms per bee for Clothianidin, 13 

### 20 nanograms per bee for Thiamethoxam; levels lower than those 

### 21 noted in EPA's Clothianidin registration documents, which report 

### 22 an LD50 more than 43.9 nanograms per bee. 

### 23 In fact Clothianidin and Thiamethoxam are only 

### 24 second in toxicity to Imidacloprid with a LD50 17.9 nanograms 

### 25 per bee. 

### 26 Studies have also reported that bees exposed

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1 to sublethal doses of pesticides are highly susceptible to 

### 2 pathogens that lead to their decline. A 2012 study by USDA 

### 3 researchers discovered that newly emerged bees exposed to 

### 4 sublethal levels of Imidacloprid during larval development and 

### 5 indirectly from brood food from nurse bees had higher levels of 

### 6 the gut parasite Nosema, which is known to adversely affect 

### 7 colony health. 

### 8 Systemic uses on corn seeds means that 

### 9 pesticide residues are translocated throughout the plant to 

### 10 pollen and nectar. Pesticide-intensive corn cultivation, which 

### 11 accounts for 80 million acres of land planted in the U.S. 

### 12 According to Krupke et al., the application 

### 13 rates for Clothianidin on corn can range from 0.25 to 1.25 mg 

### 14 per kernel. A single kernel therefore contains several orders 

### 15 of magnitude of active ingredient more than the published LD50 

### 16 values for honey bees. The translocation of residues throughout 

### 17 the plant means that pollen and nectar are highly contaminated. 

### 18 19 Similarly, Clothianidin has been found in or 

### 20 on nearby plants like dandelions resulting from translocation of 

### 21 residues from the soil to the flower, from surface contamination 

### 22 of the flowers from airborne particles, or from a combination of 

### 23 both factors. 

### 24 The panel, when making its recommendations to 

### 25 the EPA, must always keep in mind that bees are exposed to 

### 26 chemical residues through multiple routes of exposures. Bees

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1 are exposed to pesticides via foliar and systemic treatments 

### 2 when they are pollinating flowering crops, and pesticide drift 

### 3 from surrounding areas. 

### 4 They are also exposed to residues from wild 

### 5 plants contaminated through cross-pollination with corn 

### 6 systemically treated with pesticide. Bees are also exposed to 

### 7 residues in plant exhaust material produced during the planting 

### 8 of treated seed. Guttated water of seed-treated plants, which 

### 9 provides a source of water for bees, is also a source of 

### 10 contamination and exposure. 

### 11 Similarly, bees can be exposed to multiple 

### 12 pesticides at any one time while foraging. It is now accepted 

### 13 and supported by the scientific literature, that chemical 

### 14 mixtures can pose greater health hazards in combination. In 

### 15 fact, research by scientists at the University of Florida linked 

### 16 bee decline to larval exposure to a cocktail of pesticides. 

### 17 Unfortunately, EPA's risk assessment process does not 

### 18 accommodate chemical mixtures, which means that synergistic 

### 19 effects continue to go unevaluated. 

### 20 Pollinator field studies must therefore 

### 21 reflect real world events. Study designs should be based on the 

### 22 planting practices of the crop treated with pesticide, including 

### 23 evaluating the impacts of farm equipment that may result in 

### 24 airborne residues. 

### 25 Field sites must be large enough to 

### 26 accommodate the distances travelled by foraging bees, and hive

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1 entrances should be placed in a location to ensure bees will 

### 2 sample the field site. Life-cycle studies should also be a 

### 3 requirement. It is also important to note that non-Apis bees, 

### 4 in addition to pollen and nectar, are exposed to soil and tree 

### 5 pesticide residues. 

### 6 EPA states that the protection goals for honey 

### 7 bees include protecting pollination services, honey production 

### 8 and bee diversity, with the endpoints being acute lethality, 

### 9 survival, growth and the reproduction of the colony. 

### 10 For years, the science has shown that exposure 

### 11 to pesticides is detrimental to bees, and that levels not 

### 12 immediately lethal to worker adults may cause significant hidden 

### 13 damage to colonies. We hope the panel places an emphasis on a 

### 14 reliance on broadly representative, multiyear, controlled field 

### 15 tests, to provide realistic data points from which to judge a 

### 16 pesticide's impacts on honey bees. 

### 17 It is imperative that the EPA has available 

### 18 all the relevant data before any pesticide product can be 

### 19 registered and introduced into the environment. It is 

### 20 unacceptable that EPA allows pesticides to be sold and 

### 21 distributed, or expands uses of existing registrations without 

### 22 knowing how these pesticides would impact bees, other 

### 23 pollinators and essential nontarget organisms. 

### 24 In closing, we wish to impress upon the panel 

### 25 the importance for finalizing a uniform, comprehensive protocol 

### 26 that aligns with the pollinator protection goals stated by the

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1 Agency. 

### 2 Again, thank you for this opportunity to 

### 3 comment and Beyond Pesticides has already submitted written 

### 4 comments to the docket with a list of relevant scientific 

### 5 studies for the Panel's consideration. Thank you. 

### 6 **DR. DANIEL SCHLENK:**Thanks for your comments. 

### 7 Any questions from the panel? Okay. Thank you very much. 

### 8 Okay. Mr. Doan, we will get you next here. 

### 9 **MR. JAMES DOAN:**Good morning. My name is 

### 10 James Doan, and I live in Hamlin, New York. I own and operate 

### 11 Doan Family Farms, an apiary where we keep 2000 hives of honey 

### 12 bees. Our apiary places 1600 hives in fruit orchards in the 

### 13 spring and we place another 1000 hives in vegetable crops in the 

### 14 summer. Our hives travel south to Fort Meade, Florida for the 

### 15 winter, and we place our bees in orange groves in the spring for 

### 16 orange honey production. 

### 17 Beekeeping has been in my family for 57 years. 

### 18 Myself - I have kept bees for 44 years. My son, Ben, has also 

### 19 recently joined the family business and will be a fourth 

### 20 generation beekeeper. I come here today to talk about a problem 

### 21 that we have seen this year in our industry and that I feel must 

### 22 be fixed. 

### 23 In May of this year 2012, I saw our apiary's 

### 24 first heavy bee kill since 1987. Our honey bees that were hit 

### 25 then were with a chemical called methyl parathion. The 

### 26 Department of Environmental Conservation that year documented

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1 and did tests on the hives showing the very high loss at that 

### 2 point due to methyl parathion. What we saw this year was very 

### 3 much like methyl parathion. We haven't seen a bill kill like 

### 4 this since that point 25 years ago, and I really don't want to 

### 5 see damage like this again. 

### 6 We had placed bees in an apple pollination 

### 7 location in Red Jacket, Geneva in March of this year. We were 

### 8 called to remove them, and when we got there this spring, the 

### 9 bees were dead, crawling on the ground in front of the hives. 

### 10 These are huge hives of bees, single stories, nine frames a 

### 11 brood, two boxes of honey on them. 

### 12 We removed them, took them back to my farm. I 

### 13 called the DC, which is our Department of Conservation that next 

### 14 morning. They refused to come out. The lady at Region 8 

### 15 Department of Environmental Conservation told me they had no 

### 16 budgetary money to come and look at any bee kills. I will side 

### 17 note here - if I shot a bear, they would have been out in about 

### 18 15 minutes. Bees are not a priority apparently, in New York, to 

### 19 be tested. 

### 20 So I called our New York State Inspection 

### 21 Service. The New York State Inspection Service said, yeah we 

### 22 will come out, at least we can check for mites, we can check for 

### 23 nosema, and we will pull samples, because we knew at this point 

### 24 we had to do something to follow through. We had a good live 

### 25 bee kill. The bees were still coming out the next morning just 

### 26 piling out of these hives.

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1 When he got there, the bee inspector, Bob 

### 2 Duncan, opened the hives up. They had two boxes of honey capped 

### 3 off on them. There was eight to nine frames a brood in a 

### 4 single-story colony bees there were about two to three frames of 

### 5 bees left at that point. 

### 6 He collected up wiggling bees on the fronts of 

### 7 the hives and in the grass in front of the hives, put them into 

### 8 bags, froze them, and he took them to his place and held them. 

### 9 At that point, we didn't know quite where to 

### 10 send them. So, I called Maryann Frazier down to Penn State. 

### 11 She said that they had a program where they would pay half the 

### 12 cost of getting the samples run. We sent them to her. She sent 

### 13 them on to the USDA bee lab down in Gastonia. 

### 14 They ran them through as quickly as possible 

### 15 because, as I said, we've never seen anything like this at the 

### 16 same time -- and you will see, I have an illustration, there is 

### 17 a picture here on the forth page. That is what the bees looked 

### 18 like the day we got them back to our place out front. That is 

### 19 after already they have been at least one or two days into bee 

### 20 kill on that. 

### 21 I called the farmer the next morning also. I 

### 22 wanted to find out what the hell he had done to my bees. He 

### 23 said he had not sprayed yet. They were waiting for us to get 

### 24 the bees out. The only thing he told us was the guys around him 

### 25 were planting corn at that point. 

### 26 So, we took that information, you know, along

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1 with pulling the samples and that. What we came back with is as 

### 2 you can see on my next page is they checked for mites of course. 

### 3 Our mite levels range from 9 to 3 to 18. They weren't 

### 4 extremely high, high enough where we probably had been in 

### 5 treating, but we wouldn't have to worry about that. 

### 6 Nosema samples - the first sample they pulled 

### 7 came back negative, so there wasn't nosema on that one. The 

### 8 second one came back with 3,600,000 spores, which is still, in 

### 9 the bee industry today, is not considered a high level of nosema 

### 10 anymore. I know men that are running 50 to 75 million spore 

### 11 counts and still not suffering huge losses. 

### 12 We got back our results and on the page 

### 13 following the nosema report, you will see what things were found 

### 14 in the pollen. The first page is the pollen. You know, and 

### 15 certainly some of the things we found, Clothianidin was one of 

### 16 them, Captan in the pollen at extremely high levels, but in the 

### 17 bees themselves, Clothianidin was the thing that showed up. 

### 18 I mean, there are a few other things here, but 

### 19 nothing that really strikes you like Clothianidin with all the 

### 20 bees sitting there in those orchards with corn being planted 

### 21 around them. 

### 22 So, you can take your assumptions for what 

### 23 they are as far as how the bees got into Clothianidin. It was 

### 24 an apple orchard with dandelions. They shouldn't have been out 

### 25 working in any corn fields at that point, and I don't know if 

### 26 they had been flying through dust. But that brings us to the

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1 next problem. 

### 2 We lost all those hives, all 148. 

### 3 Consequently, we tried to put splits and nukes back into those. 

### 4 Those all died also. So, it was useless for us to waste our 

### 5 time in doing that. We might as well burn the equipment and 

### 6 that. That is an economic huge loss to me, 148 hives out of 

### 7 2000 is a lot of money. 

### 8 The next thing I want to show you is in June 

### 9 on the last page, you will see a big color photo, and those are 

### 10 mostly all drones. I have about 47 bee arches that had an inch 

### 11 thick pile of drones a foot in front of the hives. I haven't 

### 12 gotten the test results back, but I can tell you that is not 

### 13 nosema and its not rural mites. 

### 14 So, I don't know what chemical it is, but I 

### 15 have never seen it. I can tell you that I've talked to guys 

### 16 across the country, nobody has ever seen this. The genitalia 

### 17 are out on these bees, some of these drones are only days old. 

### 18 This is not normal. It's something that you need to be aware 

### 19 of. There are chemicals out here that are doing really bad 

### 20 things in bee arches. 

### 21 Consequently what happened after this was we 

### 22 had no matings on our queens, so all the hives that swarmed - 

### 23 drone layers or queen-less hives. My loss this summer is 

### 24 running over 20 percent and that is after we put in 1000 queens 

### 25 on a 2000 hive operation over the summer. This is pretty 

### 26 dramatic stuff.

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1 You know, I didn't put it in here, but we are 

### 2 probably re-queening 100 percent of our hives over the course of 

### 3 a year, and I'm not alone in what is going on. It's everyone 

### 4 replacing huge amounts of queens in their operations just to 

### 5 maintain the hives they have right now. And we are still losing 

### 6 bees. 

### 7 I mean, don't get me wrong, there are other 

### 8 problems out there, but when I see bees laying in front of hives 

### 9 like this, that is not those problems. 

### 10 You know, I know there is a big problem - I 

### 11 want to go back to some of the other problems I see are the LD50 

### 12 of Clothianidin. I don't know who runs the test exactly beside 

### 13 the applicant, but in the tests that they do on LD50, do they 

### 14 specify the age of the bees they are working with? You know, 

### 15 contact versus oral - we've got so many multitudes of variables 

### 16 with these things. I am not sure the LD50 - I think it needs to 

### 17 be re-looked at. 

### 18 Then we talk about synergy, stacking products. 

### 19 You know, I talked to one of my corn friends, he has seven 

### 20 different products that go on his corn seed when they go in the 

### 21 ground. What does that do? I mean, when guys are spraying 

### 22 apple orchards, they don't just put in one product. They are 

### 23 mixing two or three at the same time. What's that doing and 

### 24 what do those things together? Is that being looked at? 

### 25 You know, another issue is the tolerance 

### 26 level. I don't know how many of you are aware of it, but there

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1 are only three products that have a tolerance level at honey, 

### 2 fluvalinate, coumaphos and fenpromexite. That's it. 

### 3 All this other stuff that we find in bee 

### 4 hives, it isn't supposed to be there; and I can tell you today 

### 5 the push is in the United States, they want pollen and honey in 

### 6 those containers of honey. They want beeswax. We know what's 

### 7 in beeswax guys and ladies. We know what's in pollen. I mean, 

### 8 this isn't rocket science. 

### 9 I mean, one of these days, there's gonna be a 

### 10 big issue, who's is going to pay for that cleanup and mess? 

### 11 What I am going to do with barrels of honey that I can't do 

### 12 anything with? I can't sell them to anybody and I have to take 

### 13 them to the HAZMAT? No. We need tolerance levels for all of 

### 14 these chemicals that are out there today in honey. Captan, you 

### 15 know, all of them. We just have to have them. You have them in 

### 16 every other food out there. We have to have them. 

### 17 You know, EPA's job is to protect the citizens 

### 18 of this country from unregulated pollution in our environment. 

### 19 As a citizen, I don't feel I'm protected right now. I have 

### 20 honey bees that are being killed by products that are not 

### 21 supposed to kill bees. I have pesticides that cause my cost to 

### 22 go up the tree. And because the results are increased disease 

### 23 levels, they threaten my livelihood. War has been started over 

### 24 less and I feel as though I'm in one; a war to save the bee and 

### 25 me. 

### 26 Please look at the way data I gathered on

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1 incidents. Please look at LD50s, please work on getting 

### 2 tolerance levels in honey for chemicals, and please help us save 

### 3 the bees. Thank you. 

### 4 **DR. DANIEL SCHLENK:**Thanks for your comments. 

### 5 Any questions for Mr. Doan? Okay. Thanks a lot. Appreciate 

### 6 it. Oh you have one, Dr. Pistorius? 

### 7 **MR. JENS PISTORIUS:**Yes, Pistorius. I was 

### 8 wondering, maybe I missed it during the talk. You said that one 

### 9 incident where you provided pictures that a lot of drones were 

### 10 found dead. 

### 11 **MR. JAMES DOAN:**Right. 

### 12 **MR. JENS PISTORIOUS:**Was that the colony that 

### 13 the relation of female bees to drones were changed and do you 

### 14 think that a lot of drones were affected only? 

### 15 **MR. JAMES DOAN:**It seemed like only drones. 

### 16 In that picture, there are some workers, but predominantly it 

### 17 was drones -- all the drones in the hives and every hive it was 

### 18 like that. 

### 19 **MR. JENS PISTORIOUS:**Okay, thank you. 

### 20 **DR. DANIEL SCHLENK:**Dr. Fefferman? 

### 21 **DR. NINA FEFFERMAN:**Sorry, this is just 

### 22 scientific curiosity, do you have any impression from that? 

### 23 Were the drones trying to get back in and being forced out, or 

### 24 were they just flying out? 

### 25 **MR. JAMES DOAN:**No, we were in a major honey 

### 26 flow at that point. Those drones were not kicked out. They

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1 were crawling in the grass in front of the hives dying. We are 

### 2 still waiting on the results back from that. I have 50 bags of 

### 3 samples to run. We have lots of samples collected. It is a 

### 4 matter of who can pay for all these to be run. Right now, 

### 5 beekeepers are paying the cost of running all these things. Yes 

### 6 sir? 

### 7 **DR. GREG HUNT:**So, in those same hives 

### 8 earlier in the year, you had a bee kill. 

### 9 **MR. JAMES DOAN:**Right. 

### 10 **DR. GREG HUNT:**And now this is June. 

### 11 **MR. JAMES DOAN:**Right. 

### 12 **DR. GREG HUNT:**Okay. I just wanted to get 

### 13 the background. 

### 14 **MR. JAMES DOAN:**Now those 148 hives 

### 15 completing were died after the second time. We just push that 

### 16 stuff off to the side and dump all the frames out and burned it 

### 17 up. I couldn't - I was - I couldn't deal with them. There was 

### 18 nothing there - I wasn't going to put anymore bees into it. 

### 19 **DR. GREG HUNT:**So these drones were in other 

### 20 hives? 

### 21 **MR. JAMES DOAN:**Yes sir, in every yard, every 

### 22 hive in every yard. 

### 23 **DR. GREG HUNT:**In every yard. The crops 

### 24 around were similar? 

### 25 **MR. JAMES DOAN:**Well, our area is 

### 26 predominantly corn, soybeans, cabbage, apples, it has dairy,

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1 it's a mixture of everything. 

### 2 **DR. GREG HUNT:**Okay. Thanks. 

### 3 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 4 **DR. MAY BERENBAUM:**Did you have to pay for 

### 5 the Gastonia analysis? 

### 6 **MR. JAMES DOAN:**Yes. 

### 7 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 8 **DR. JEFF PETTIS:**Back to the incident 

### 9 reporting, I realize that state by state, there's a lot of 

### 10 variation in that. The EPA, I believe, has set up a way of 

### 11 directly reporting, have you tried that at all? 

### 12 **MR. JAMES DOAN:**I did do that once I found 

### 13 out about it, but there were a lot of beekeepers in New York 

### 14 state that didn't know about it at the time and a lot of 

### 15 beekeepers didn't have samples pulled. I've talked to people 

### 16 from all across the country that had very difficult time trying 

### 17 to get somebody from their state to come up and pull samples. I 

### 18 have talked to the Canadian beekeepers and they're telling me 

### 19 very similar stories to what I'm telling you, other than they 

### 20 got a lot of help out of their ministry to pull samples. Thank 

### 21 you very much. 

### 22 **DR. DANIEL SCHLENK:**Any other questions? 

### 23 Okay. Our next presenter is Dr. Mike Beevers from California 

### 24 Agriculture Research. 

### 25 **DR. MIKE BEEVERS:**Can you hear me? Yeah, my 

### 26 name is Mike Beevers. I'm a research director for California

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1 Agricultural Research, and we're a contract research 

### 2 organization sometimes called a private lab. We've done 

### 3 regulatory honey bee studies for about 26 years. I'm just here 

### 4 to share a little bit of perspective of the people who are doing 

### 5 some of the work right now and may be doing a lot of the 

### 6 research in the future, just to let you know kind of our 

### 7 understanding of this. 

### 8 The subject here is Potential Changes in 

### 9 Regulatory Research Requirements for Pollinators. Regulatory 

### 10 Research is the bailiwick of the contract research organization. 

### 11 We perform scientific studies for submittal to EPA or similar 

### 12 government entities by manufacturers of agricultural products. 

### 13 Our goal is to provide independence, objectivity and reliability 

### 14 in the studies. 

### 15 Our studies produce data to answer very 

### 16 specific risk assessment concerns regarding pesticides held by 

### 17 government regulatory bodies and our data is confidential. I am 

### 18 trying to just describe this as regulatory research. 

### 19 Exploratory research is something entirely different and seeks 

### 20 to answer more broad ranging hypothetical questions. Most of 

### 21 this work is peer reviewed and is not under GLP and it is done 

### 22 at the universities. Obviously, this work is very important 

### 23 going forward with bees, the university work especially. 

### 24 Contract research organizations assure the 

### 25 public of their independence by following good laboratory 

### 26 practices as established by the EPA.

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1 I was going to ask the Panel, are you familiar 

### 2 with the Good Laboratory Practice Provisions? Just to let you 

### 3 know, we do function under these. These standards are the 

### 4 cornerstone of most contract research organizations, and there 

### 5 are specific standards regarding the data, the recording of 

### 6 data, raw data, protocols, study director, legal 

### 7 responsibilities, quality assurance, in-life audits, data 

### 8 auditing of final reports, archiving, standard operating 

### 9 procedures, short-notice inspections by the enforcement branch 

### 10 of the EPA out of Denver. 

### 11 The purpose of these guidelines is to assure 

### 12 that the agencies, when they get data from our labs, are getting 

### 13 something they can rely on that they can go back years later and 

### 14 produced to defend their decisions. 

### 15 A very simple example of GLP Practice would be 

### 16 placing a sample of nectar or pollen in a freezer. That freezer 

### 17 has to have a maintenance log. The sample must be logged into 

### 18 the freezer. The freezer must have 24/7 temperature monitoring. 

### 19 It has to have alarms, probably a generator backup for power 

### 20 failures. When the sample is removed, it must be logged out, 

### 21 chain of custody, so forth and so on. I am just trying to 

### 22 indicate that there is a lot of work that goes into these 

### 23 studies to make them valid. 

### 24 Noncompliance with these laws can result in 

### 25 fines and imprisonment. There are people on contract labs who 

### 26 have actually gone to prison. I probably don't need to explain

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1 that if working under these circumstances, people take the data 

### 2 very, very seriously. I just would like to communicate that to 

### 3 you guys, and I appreciate you listening on this. 

### 4 The lab's interest is in alignment with the 

### 5 Agency's interest in terms of data quality and objectivity. The 

### 6 people in the labs are interested in learning the newest methods 

### 7 and learning to implement them. 

### 8 I would just like to say that, you know, the 

### 9 people in these labs, and I'm speaking for myself, would like to 

### 10 be a resource in the future as part of this and we are operating 

### 11 under a very strict guideline. 

### 12 Finally, if anyone knows of any problems with 

### 13 labs -- some of these labs have been criticized in the past -- I 

### 14 would be interested in talking with them at some point. That is 

### 15 all I have to say, thank you. 

### 16 **DR. DANIEL SCHLENK:**Thank you. Any questions 

### 17 for Dr. Beevers? Dr. Pettis? 

### 18 **DR. JEFF PETTIS:**There has been a lot of work 

### 19 in Tier I testing on Apis mellifera. Have you done any testing 

### 20 with other species of pollinators at all? 

### 21 **DR. MIKE BEEVERS:**Not successfully, no. The 

### 22 non-Apis issue is very tricky for, as I understand it, for 

### 23 getting predictable, replicable tests. The honey bee is just a 

### 24 wonderful organism to work with in comparison with those. 

### 25 **DR. JEFF PETTIS:**I wouldn't argue about the 

### 26 honey bee, but my concern is that if we were to move to other

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1 species testing, what would be reliable? 

### 2 **DR. MIKE BEEVERS:**We would be - I have worked 

### 3 with leaf cutter bees. We have tried starting with bumble bees. 

### 4 Yes, that can be done and we are interested in doing it, but to 

### 5 be very honest with you, I haven't had a lot of demand for that. 

### 6 But that is an area we are interested in, and we would love to 

### 7 try. 

### 8 **DR. DANIEL SCHLENK:**Okay. Any other 

### 9 questions? Yep, Dr. Tarpy? 

### 10 **DR. DAVID TARPY:**This is Dave Tarpy, NC State 

### 11 University. Is there an analogous kind of GLP, but not for the 

### 12 L but for kind of like ecological setting that would hold up to 

### 13 that same stringency as the GLPs that you work under? 

### 14 **DR. MIKE BEEVERS:**That's an excellent 

### 15 question, and not to my knowledge. The studies in ecological 

### 16 settings are extremely complicated. We have applied the good 

### 17 laboratory practice to the data. There're a lot of things that 

### 18 would transfer there, but in terms of actually addressing some 

### 19 of the possible issues that you may have in mind, we are dealing 

### 20 with a good laboratory practice for that purpose right now. 

### 21 **DR. DAVID TARPY:**Thank you. 

### 22 **DR. DANIEL SCHLENK:**Any other questions? Dr. 

### 23 Pistorius? 

### 24 **MR. JENS PISTORIUS:**Assuming you have a lot 

### 25 of contact with other GLP laboratories in the U.S. too, my 

### 26 question is have you established in vitro test method yet as a

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1 routine method and with acceptable control mortality, and are 

### 2 you able to do to tests longer than day 7? 

### 3 **DR. MIKE BEEVERS:**The answer to that is 

### 4 essentially no. The in vitro test has been tested -- no from me 

### 5 personally. Some of the other labs may have developed that, but 

### 6 I'm not aware of. We have ring tested one in vitro test, but my 

### 7 understanding is it's still in development. 

### 8 **DR. DANIEL SCHLENK:**Any other questions? 

### 9 Okay. Thank you, Dr. Beevers. 

### 10 **DR. MIKE BEEVERS:**Thank you. 

### 11 **DR. DANIEL SCHLENK:**Our next presenter is Rod 

### 12 Snyder from the National Corn Growers Association. 

### 13 **MR. ROD SNYDER:**Good morning. My name is Rod 

### 14 Snyder, Director of Public Policy for The National Corn Growers. 

### 15 We appreciate the opportunity to just have a few brief remarks 

### 16 this morning. I will be providing comments on behalf of NCGA, 

### 17 which represents more than 37,000 members in 48 states and more 

### 18 than 300,000 corn farmers who contribute to state checkoff 

### 19 programs across the country. 

### 20 I want to focus just these very brief 

### 21 statements on the benefits of seed treatments since that is 

### 22 where we've been getting a lot of questions recently regarding 

### 23 the studies for their potential impacts on pollinators. 

### 24 Seed treatments are critically important to 

### 25 growers because they protect the seeds from disease and pests 

### 26 from planting through germination. Unpredictable weather

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1 patterns during the planting season could lead to the seed 

### 2 remaining in the ground for an extended period of time and 

### 3 without these products, yields would be reduced and entire 

### 4 fields may need to be replanted. 

### 5 Seed treatments lead to a higher yield corn 

### 6 crop by providing a healthier root system, particularly by 

### 7 maximizing early season plant stands. Growers also consider 

### 8 these seed treatments an important investment as they seek to 

### 9 protect and maximized high-value seed. 

### 10 Field tests for the products have demonstrated 

### 11 a 6 bushel per acre average yield increase in corn. For 

### 12 example, Clothianidin is an important neonicotinoid seed 

### 13 treatment for corn in the U.S. The product is used in 

### 14 approximately 60 percent of corn planted in this county. It is 

### 15 a valuable product as growers seek to control wireworms, black 

### 16 cutworms, white grubs and other early-season pests that attack 

### 17 corn seeds and seedlings at a period when they are most 

### 18 vulnerable. 

### 19 It is also worth noting that in no-till 

### 20 practices, no-till operations, because the soil health is 

### 21 improved, it also means additional pest pressures. So, all of 

### 22 the benefits from no-till operations could be lost if we have to 

### 23 start tilling again because of pressure on the seeds. 

### 24 The use of seed treatments reduces the 

### 25 potential human health effects for corn growers because it 

### 26 reduces the need for soil-applied sprays or granules.

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1 Environmental exposures are also reduced due to the fact that 

### 2 the seed is incorporated beneath the soil surface at planting 

### 3 time. This leads to a lower probability of exposure to birds or 

### 4 runoff to water bodies when compared to soil-applied products. 

### 5 Seed treatments actually have the effect of reducing the amount 

### 6 of active ingredient needing for effective pest control. 

### 7 Eliminating foliar applications also removes 

### 8 the concern with making extra trips across the field during a 

### 9 time of the year when rainfall and muddy soils can create major 

### 10 time constraints during planting season. Fewer trips across the 

### 11 field also leads to fewer greenhouse gas emissions from fossil 

### 12 fuels used in tractors and other farm equipment. 

### 13 While corn growers remained concerned about 

### 14 the recent declined in bee populations in the U.S. and the 

### 15 potential impact on the larger agricultural economy, studies 

### 16 suggest that there are multiple and complex causes of colony 

### 17 collapse disorder that cannot be explained by a single class of 

### 18 chemicals or agricultural practices. 

### 19 There are no demonstrated long-term effects on 

### 20 bee colonies from neonicotinoid pesticides. Furthermore, corn 

### 21 growers work diligently to minimize dust from planters and often 

### 22 establish relationships with nearby beekeepers to minimize 

### 23 exposure to pollinators. We appreciate the opportunity to 

### 24 comment today, and we urge this Panel to maintain a 

### 25 scientifically sound risk assessment process when reviewing the 

### 26 impact of seed treatments on pollinators. I am certainly happy

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1 to follow up with other information that you need. Thank you. 

### 2 **DR. DANIEL SCHLENK:**Any questions? Yes, Dr. 

### 3 Hunt? 

### 4 **DR. GREG HUNT:**Yeah, I've heard that 

### 5 undoubtedly seed treatments are good for corn yields. I've 

### 6 heard the 6-bushel an acre data before. Is that in a published 

### 7 study, and has there been a study that looked at the benefit of 

### 8 just the neonicotinoids without the other fungicides, et cetera? 

### 9 10 **MR. ROD SNYDER:**Sure, I'd have to go back and 

### 11 figure as we were piecing this together where that citation came 

### 12 from, but we can easily follow up on that. Yeah, absolutely. 

### 13 **DR. DANIEL SCHLENK:**Dr. Potter and Dr. 

### 14 Berenbaum? 

### 15 **DR. THOMAS POTTER:**Yeah, I just feel 

### 16 compelled to make a comment here because regarding your point 

### 17 that you made about increased pest pressure with no-till 

### 18 practices associated with improved soil health. I'm not sure 

### 19 what you mean by that, but I think there're probably other 

### 20 issues other than improved soil health that may cause increased 

### 21 problems with pests, probably just the fact that the soils are 

### 22 colder and wetter. 

### 23 I don't mean to make too big a point about 

### 24 that, but I think improved soil health is a really good thing. 

### 25 It is something that I've connected my career mostly to, so you 

### 26 know, I think there are other issues than soil health that may

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1 contribute to pest pressure in no-till systems, which certainly 

### 2 are important conservation system set up throughout the country. 

### 3 **MR. ROD SNYDER:**Yes, by all means. Improved 

### 4 soil health is a big part of the reason why farmers shift to 

### 5 no-till, and I'm sorry if I've left any confusion there. There 

### 6 are many, many benefits to no-till systems. Our growers are 

### 7 moving in that direction rapidly in the last 10 or 20 years. 

### 8 But you are right. Whether it is the temperature of the soil of 

### 9 what have you, there can be additional pest pressures in those 

### 10 settings. So, seed treatments allow that adaptation to no-till. 

### 11 That's just, sorry, the basic point I was trying to make. 

### 12 **DR. MAY BERENBAUM:**Wireworms and white grubs 

### 13 and black cutworms are not uniformly distributed across the 

### 14 corn-growing region and differ in their importance as economics 

### 15 pests. Do you know if there is - that 60 percent of corn 

### 16 growers who use pretreated seed - is that predominantly in 

### 17 regions where there is history of pest problems? 

### 18 **MR. ROD SNYDER:**As far as the distribution of 

### 19 where the product is used, I'd have to look that up and get back 

### 20 to you. I mean certainly it's a common product in the corn-belt 

### 21 in the midwest and upper midwest, but I can look that up for 

### 22 sure. 

### 23 **DR. DANIEL SCHLENK:**Any other questions? Dr. 

### 24 Pistorius? 

### 25 **MR. JENS PISTORIUS:**Short one. How high do 

### 26 you think is the percentage of non-tillage systems for maze

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1 cropping? 

### 2 **MR. ROD SYNDER:**I'm sorry, the question is 

### 3 how - 

### 4 **MR. JENS PISTORIUS:**How big is the portions 

### 5 of fields where there is no tillage for sowing? 

### 6 **MR. ROD SNYDER:**That is a little bit region - 

### 7 there are regional differences. I can say with certainty in the 

### 8 mid Atlantic region where there's been a lot of emphasis on 

### 9 water quality in trying to prevent runoff because of the 

### 10 Chesapeake Bay situation, recent studies have shown that close 

### 11 to 90 percent of acreage in this region has moved to either 

### 12 no-till or conservation tillage. It is a little lower in the 

### 13 midwest, but I mean, it's a very high percentage. 

### 14 **DR. DANIEL SCHLENK:**Dr. Hunt? 

### 15 **Dr. GREG HUNT:**Just real quick, I was a 

### 16 little surprised by you saying that only 60 percent of the 

### 17 growers are using treated seed. 

### 18 **MR. ROD SNYDER:**That was Clothianidin 

### 19 specifically. Other products would drive that number much 

### 20 higher. Clothianidin has a lot - there has been a lot of talk 

### 21 about that product in particular. 

### 22 **DR. GREG HUNT:**Perhaps up to 98 percent. 

### 23 **MR. ROD SNYDER:**Right. 

### 24 **DR. DANIEL SCHLENK:**Okay. Any other 

### 25 questions? Okay. 

### 26 **MR. ROD SNYDER:**All right, thanks.

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1 **DR. DANIEL SCHLENK:**Thank you. Our last 

### 2 public commenter today is Stephen McFadden from Independent 

### 3 Scientific Research Associates. And Mr. McFadden, if I could 

### 4 have you to about a 5 to 10 minute range, is that appropriate? 

### 5 Thanks. 

### 6 **MR. STEPHEN MCFADDEN:**Hello. I'm from 

### 7 Midland, Texas and after all the academic discussion, I want to 

### 8 point out where the rubber hits to road. We recently had a 

### 9 larger urban aerial spraying in Dallas, Texas. The DFW 

### 10 Metroplex is 6.5 million people. It's the largest metropolitan 

### 11 area in the south. The city of Dallas is 300 square miles. In 

### 12 2003, we beat the West Nile virus using an IPM program put 

### 13 together by former EPA'er, Bill Curry. 

### 14 The West Nile virus virulence has a feedback 

### 15 term where if you have a bad year, the next year a lot of birds 

### 16 are immune. And so, you have a good year the next year. But 

### 17 after nine years, then the birds aren't immune. 

### 18 So, after nobody was watching the store for 

### 19 2010 and 2011, in 2012 we had the worst outbreak probably in the 

### 20 U.S. ever. They aerially sprayed 1150 square miles twice, 

### 21 mostly two nights in a row, homes of maybe 2 million people. 

### 22 They used Clarke Duet with Sumithrin and Prallethrin from 

### 23 twin-engine planes at 300 feet at night. From 300 feet, the 

### 24 spray probably takes between 20 minutes and 2 hours to hit the 

### 25 ground, I'm guessing. So there is probably a lot of drift. 

### 26 A few organic farmers were upset. Some

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1 beekeepers were upset. I would image that a number of bees were 

### 2 upset. 

### 3 There were presumably a bunch of governmental 

### 4 affairs, three issues. One of our issues is like on the City of 

### 5 Dallas website it says that aerial spraying is safe and 

### 6 effective. EPA does not allow the term safe to be used to apply 

### 7 to pesticides, but this came from the top state health office in 

### 8 Texas who was using the term safe including on a CDC 

### 9 teleconference, and nobody confronted him on it. 

### 10 The remedy is to sticker the label of public 

### 11 health pesticides with the statement that it is a violation of 

### 12 EPA regulations and of the label to state that the pesticide is 

### 13 safe. 

### 14 Secondly, one resident ran an adverse effects 

### 15 hotline on the reregistration docket for Prallethrin. They 

### 16 submitted 14 cases of adverse effects. Given the number of 

### 17 instances of vomiting, they wonder that if Prallethrin, which is 

### 18 supposed to cause benign agitation of mosquitos and rapid knock 

### 19 down, if that may have some human effects such as vomiting. 

### 20 Thirdly, they hit 1150 square miles twice. 

### 21 Some people think this is the new norm. We wonder if maybe they 

### 22 should file an Environmental Impact Statement because in the EIS 

### 23 process, maybe the bees would get some representation. Thank 

### 24 you. 

### 25 **DR. DANIEL SCHLENK:**Any questions for Mr. 

### 26 McFadden before he leaves. Would you like to answer any

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1 questions? You don't have to. Dr. James? 

### 2 **DR. ROSALIND JAMES:**Could you tell me what 

### 3 your organization is again. 

### 4 **MR. STEPHEN MCFADDEN:**Independent Scientific 

### 5 Research Advocates. 

### 6 **DR. ROSALIND JAMES:**Okay, who is that? 

### 7 **MR. STEPHEN MCFADDEN:**Well, it's me, myself 

### 8 and I, and I also run the website dalcap.org. And I work with 

### 9 several people in the Dallas Fort Worth area. In 2002 and 2003, 

### 10 I was living in Dallas, and we got on the city council to try to 

### 11 put together IPM program for the West Nile virus. So, in 2002 

### 12 and 2003, I was living there on a daily basis. 

### 13 This year, I cancelled my trip to Dallas when 

### 14 I saw aerial spraying listed, but I followed basically every 

### 15 news article on the internet and I work closely with the 

### 16 activists there. 

### 17 It's just like all of a sudden, government 

### 18 screwed up, nobody is watching the store and all of the sudden, 

### 19 they tell the public we are going to be spraying by air in a 

### 20 couple of days and it's going to hit 1000 square miles. 

### 21 The bees don't like it, we don't like it. 

### 22 They claim its safe in the all the press and we know that that 

### 23 isn't supported that the claim of safety fails to consider 

### 24 individual factors such as genetics, health history, et cetera. 

### 25 So the term safe is conclusive when you can't make a conclusory 

### 26 term about 2 million people. Thank you.

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1 **DR. ROSALIND JAMES:**Thank you. 

### 2 **DR. DANIEL SCHLENK:**Okay. At this point in 

### 3 time, our time of public commenting is closed. I believe so. 

### 4 So, before lunch if we could, if we can have the Agency come up 

### 5 again, and if the Panel has any questions related to the 

### 6 presentations that were made yesterday, you've had the evening 

### 7 to sort of mull over those presentations and your question 

### 8 groups discuss a few of those. If you have any particular 

### 9 questions to any particular representatives here, now is the 

### 10 time to do it before we get into our panel question 

### 11 deliberations. So, Dr. Berenbaum? 

### 12 **DR. MAY BERENBAUM:**I just have a question, 

### 13 don't even know to whom it should be directed, but it is for 

### 14 general information and I apologize if everyone else knows this, 

### 15 but I don't. Incident reporting - who is, how is that - we've 

### 16 heard there is state level, federal level, what agency, who has 

### 17 oversight, how is it regulated? 

### 18 **DR. THOMAS STEEGER:**The only entity that's 

### 19 required to report incidents to the Agency is the Regulated 

### 20 Community Registrants themselves under FIFRA 6A2. The Agency 

### 21 provides funding to states who are the - we refer to them as the 

### 22 State Lead Agencies. They have primacy for enforcing labels and 

### 23 they are responsible for investigating incidents that are 

### 24 reported to them. 

### 25 The Agency is aware that many beekeepers feel 

### 26 that for a variety of reasons incident reports are not making it

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1 back to the Agency. The Agency makes clear in its assessments 

### 2 that the absence if incident reports cannot be construed as the 

### 3 absence of incidents. 

### 4 So, understanding that there have been 

### 5 breakdowns in communication and that the number of incidents is 

### 6 potentially being under reported, there is reluctance by many 

### 7 beekeepers to report incidents because of fear of offending 

### 8 growers that they depend on for pollination services and for 

### 9 honey production. And, there is actually fear of retribution 

### 10 that bee colonies will be destroyed. 

### 11 That is very unfortunate. But understanding 

### 12 that there are impediments to reporting incidents, the Agency 

### 13 has established other means for individuals to contact the 

### 14 Agency directly through the National Pesticide Information 

### 15 Center, and they can contact us through beekill@epa.gov. I 

### 16 monitor the portal for the beekill@epa.gov. 

### 17 We did have the opportunity to record Jim 

### 18 Doan's bee kill incidents, and we were in contact with the state 

### 19 of New York through our regional offices to encourage them to 

### 20 investigate those incidents. 

### 21 **DR. DANIEL SCHLENK:**Any other questions? Dr. 

### 22 Potter? 

### 23 **DR. THOMAS POTTER:**Yes, I have a question 

### 24 about the data that was summarized in the document describing 

### 25 pesticide concentrations in pollen and nectar. And my question 

### 26 is does that represent the entire set of information that you

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1 utilized in the process of establishing a dietary exposure 

### 2 estimate using T-REX? 

### 3 **MS. KRIS GARBER:**This is Kris Garber. We 

### 4 used all the available information from the open literature and 

### 5 unpublished registrant submitted studies that we are aware of. 

### 6 We did some literature searches, and we searched our database of 

### 7 submitted studies. 

### 8 We did exclude some studies from the open 

### 9 literature that didn't provide application rate information 

### 10 because it was really important for us to be able to link the 

### 11 mass of the pesticide that was put on the field so that, you 

### 12 know mass that was observed in the different samples, especially 

### 13 for the T-REX and for the Briggs' analysis. So, there are some 

### 14 studies in the literature that we excluded based on application 

### 15 rate. 

### 16 **DR. THOMAS POTTER:**I got the impression, and 

### 17 maybe again, that is why I'm following up on this, that there 

### 18 were registrant submitted studies that contained information on 

### 19 active ingredient concentrations in pollen and nectar that were 

### 20 utilized. So, what you're telling me is that is not the case 

### 21 other than what you describe in the White Paper? 

### 22 **MS. KRIS GARBER:**What I'm saying is that 

### 23 there were registrant studies that had concentrations in pollen 

### 24 and nectar, and those are included in the White Paper and they 

### 25 are summarized in appendices 3 and 4. 

### 26 **DR. THOMAS POTTER:**Right, and again, I will

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1 just make a point here that I think there are to me a 

### 2 surprisingly small number of measurements that are available to 

### 3 use in making assessments about the dietary exposure here. You 

### 4 know, I think that is critically important. We will get into 

### 5 that this afternoon. Maybe I'm jumping the gun here a bit, but 

### 6 I wanted to confirm what data you had utilized in your 

### 7 assessments. 

### 8 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 9 **DR. JEFF PETTIS:**Jeff Pettis. Kind of 

### 10 following up on that, in looking at the consumption rate of 

### 11 nectar versus pollen, I just had a feeling that the pollen 

### 12 consumption rate seemed abnormally low. Can you comment on the 

### 13 calculations of the average bee consuming nectar versus pollen? 

### 14 15 **MS. KRIS GARBER:**Well, we - both of the 

### 16 consumption rates for pollen and nectar were empirically based. 

### 17 The pollen consumption rates are based on direct counts of 

### 18 pollen grains in the guts of bees, including adults and larvae. 

### 19 So, I think those are some pretty confident measures because 

### 20 they were actually able to, you know, count how many pollen 

### 21 grains were in each bee. 

### 22 Now the nectar consumption rates are not based 

### 23 on direct measures of how much a bee eats, either nectar or 

### 24 honey. So we had to calculate it based on the energetic 

### 25 requirements of the bees in terms of how much sugar they may 

### 26 need based on their different tasks. So the nature of the

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1 estimate is different. 

### 2 Perhaps that could lead to some differences in 

### 3 the magnitude of those two. But for a nectar forager that 

### 4 spends a lot of its time flying, it makes sense that it would 

### 5 need a lot of sugar to, you know, fulfil its task. 

### 6 **DR. DANIEL SCHLENK:**Dr. James? 

### 7 **DR. ROSALIND JAMES:**This is Rosalind James. 

### 8 I have a follow-up question to that. Later today we will 

### 9 probably talk some more about this difference between the pollen 

### 10 and nectar in the EPA assessment for Tier I. In some ways, it 

### 11 wouldn't matter because you are essentially saying that the 

### 12 concentration of pesticide is the same in both, and yet some of 

### 13 the commenters today were saying pesticide concentration in 

### 14 pollen is quite a bit higher than in nectar. So, this is an 

### 15 important question. 

### 16 **MS. KRIS GARBER:**When we were coming up with 

### 17 the Tier I method for foliar spray applications, that's based on 

### 18 the tall grass residue of the surrogate for pollen and nectar, 

### 19 we did actually consider applying some kind of adjustment 

### 20 factor, which was something that was brought up in the public 

### 21 comments; but there were some studies where - and I think that 

### 22 one of them is Choudhary et al., which is summarized -- 

### 23 Choudhary and Sharma I think are the two authors. There were 

### 24 actually some cases where the concentrations in the pollen and 

### 25 the nectar were equivalent. 

### 26 So, we thought that you couldn't necessarily

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1 assume that the concentrations are always going to be less in 

### 2 nectar, although it is true that there are several studies where 

### 3 that is the case. So, that's why we decided to assume that 

### 4 nectar and pollen were equivalent and that, you know, like Dr. 

### 5 Potter had pointed out, there isn't a lot of data available for 

### 6 pollen and nectar. So, we didn't think that we had enough 

### 7 information to necessarily apply an appropriate adjustment 

### 8 factor. 

### 9 **DR. DANIEL SCHLENK:**Go ahead. 

### 10 **MR. KEITH SAPPINGTON:**Keith Sappington, EFED. 

### 11 I just want to follow up with one point on those data that they 

### 12 represent day zero values. So values, you know, measured 

### 13 immediately after application or within 24 hours, so that is 

### 14 something to keep in mind with the foliar applications. 

### 15 The other thing is we notice some differences 

### 16 across plant species. You might think that the availability of 

### 17 the nectaree to receive the spray droplets would be a factor, so 

### 18 different plants and different physiologies associated with the 

### 19 nectarees could come into play. So we looked at this, but we 

### 20 felt that the range of, basically the variability around the 

### 21 pollen to nectar ratio was too large to proposed as a method. 

### 22 **DR. DANIEL SCHLENK:**Dr. Fefferman? 

### 23 **DR. NINA FEFFERMAN:**Hi thanks. Along the 

### 24 same lines, I was actually - I like very much the sort of 

### 25 energetic analysis of the need for the workers for nectar 

### 26 consumption and the absence of their being a measurement for

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1 that. I was wondering of the similar logic couldn't be applied 

### 2 to the protein needs of egg-laying queens in order to establish 

### 3 a rate separate for consumption there. 

### 4 **MS. KRIS GARBER:**I think that's an excellent 

### 5 point and we could definitely consider that. 

### 6 **DR. DANIEL SCHLENK:**Dr. Pistorius? 

### 7 **MR. JENS PISTORIUS:**I have question. Would 

### 8 the EPA be in charge and in power to implicate risk mitigation 

### 9 measures of beyond the action of application of pesticides and 

### 10 especially, for instance, concerning seed treatment facilities, 

### 11 prescribing a certain seed quality level? And in addition to 

### 12 that, for instance like that for non-tilling systems, that there 

### 13 has to be a herbicide application or removal of flower plants 

### 14 before application - is that possible? And machine technique as 

### 15 well, so you could prescribe use of deflectors? 

### 16 **DR. THOMAS STEEGER:**The EPA has been working 

### 17 with the seed treatment organizations and with the seed 

### 18 equipment manufacturers as well as the registrants to develop 

### 19 that very type of mitigation. The label though, is specific to 

### 20 how the product is applied as opposed to how it might be 

### 21 distributed in plantars. 

### 22 So, it is very difficult from my understanding 

### 23 and out my realm for speaking to this issue, but we feel that 

### 24 the best means of mitigating those potential effects is through 

### 25 best management practices that would be imposed by industry. 

### 26 **DR. DANIEL SCHLENK:**Dr. Hunt?

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1 **DR. GREG HUNT:**I want to get back to - this 

### 2 kind of relates to the previous question in a way. I want to 

### 3 get back to the pollen, the amount of pesticide in pollen, 

### 4 specifically Clothianidin; the White Paper cites our Krupke et 

### 5 al. paper as 3.9 parts per billion in the corn pollen that we 

### 6 analyzed. But we also took pollen from pollen traps on hives 

### 7 and found up to 88 parts per billion of Clothianidin. And this 

### 8 was at a time when corn was not being planted. 

### 9 So, my hypothesis is that this is from talc 

### 10 dust that had remained on flowers for a while because that kind 

### 11 of level, we just don't find it in corn pollen that has been 

### 12 treated, although the bees were bringing in up to 80 percent 

### 13 corn pollen in the sampling. 

### 14 **DR. DANIEL SCHLENK:**So, is this a question 

### 15 for the Agency or a comment? 

### 16 **DR. GREG HUNT:**I guess this is a comment. 

### 17 **DR. DANIEL SCHLENK:**Yeah, let's hold the 

### 18 comments until later. Okay? 

### 19 **DR. GREG HUNT:**All right, sorry. 

### 20 **DR. DANIEL SCHLENK:**These are questions the 

### 21 Agency that will help you in your comments later on. Okay? 

### 22 That's what this time is for. Any other questions? Okay. 

### 23 Thanks a lot. Let's break for lunch. We will come back at 

### 24 12:50, and read in the questions at that time. Thanks. 

### 25 (WHEREUPON lunch was taken) 

### 26 **DR. DANIEL SCHLENK:**Question one - and just

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1 to save a little bit of time, who's going to be reading the 

### 2 question in? Who is planning to read the question in? Okay. 

### 3 You just want to read the letter (a) instead of the whole thing. 

### 4 I think that part will be sufficient instead of reading the - 

### 5 so the sub-letters basically throughout I think will be good. 

### 6 Great, thanks. 

### 7 **MR. THOMAS MORIARTY:**This is Tom Moriarty. I 

### 8 also understand that you want us to pause after each 

### 9 subquestion? 

### 10 **DR. DANIEL SCHLENK:**Correct. We will go 

### 11 subquestion by subquestion. 

### 12 **MR. THOMAS MORIARTY:**Just enough to take a 

### 13 breath so I can move onto the next one. 

### 14 **DR. DANIEL SCHLENK:**Okay, go ahead. 

### 15 **MR. THOMAS MORIARTY:**Tom Moriarty, U.S. EPA. 

### 16 Charge question 1, sub (a) - please comment on whether the 

### 17 assessment endpoints, population size and stability of managed 

### 18 bees, quantity and quality of hive products, and species 

### 19 richness and abundance, identified in Table 1, Section 2.2.1 of 

### 20 the White Paper, are consistent with the Agencies protection 

### 21 goals. Please include a discussion of any additional assessment 

### 22 endpoints that may be necessary to meet those protection goals. 

### 23 24 **DR. DANIEL SCHLENK:**Okay. Our lead 

### 25 discussant for this particular question is Dr. James. 

### 26 **DR. ROSALIND JAMES:**This is Rosalind James.

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1 Since I am answering the first question, I have a couple of 

### 2 general comments. They won't be long. To the group, I wanted 

### 3 to commend EPA on the excellent job in writing the White Paper. 

### 4 It's very thorough and well thought out. It's clear that a lot 

### 5 of care went into writing this. This proposal is definitely a 

### 6 step in the right direction. It is very heartening to see EPA 

### 7 taking on this task with bees. 

### 8 And I have another short comment to the group, 

### 9 the Panel here. This is a small plea on the use of language, 

### 10 and in particular, I would like to ask you not to use the words 

### 11 bees and honey bees synonymously because remember, we are 

### 12 talking about diversity and we will be talking bees other than 

### 13 honey bees. So, please say honey bees when you mean honey bees 

### 14 and when you say bees, mean more than just honey bees. It will 

### 15 be helpful for me, I know, and probably for EPA as well. 

### 16 With regard to question 1(a), essentially this 

### 17 question has to do with Table 1 on page 44. So, I am going to 

### 18 work off of the Table in my answer here. 

### 19 First we are looking at the protection goals 

### 20 that have been set by EPA and the question is whether or not the 

### 21 assessment endpoints address the protection goals. So, here I 

### 22 would like to reiterate about the bees and in question 1 for 

### 23 provisions of pollination and services. The assessment endpoint 

### 24 says managed bees. So, we wanted to make sure that EPA is clear 

### 25 that this means commercial managed bees, which means honey bees, 

### 26 bumble bees, alfalfa-leaf cutting bees and mason bees and any

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1 other bees that may become managed in the future. 

### 2 So when it says population size, we weren't 

### 3 really sure if this meant colony population size as for honey 

### 4 bees or actually truly population size, and that really it 

### 5 should probably be populations in terms of numbers of colonies. 

### 6 7 Then for production of hive products - the 

### 8 assessment endpoint is quantity and quality of hive products. 

### 9 Quantity and quality of hive products is fine endpoint, but 

### 10 probably quantity would be sufficient, unless quality really 

### 11 means pesticide-free or low pesticide residues. In this case 

### 12 quality is an important measure we will see in the next 

### 13 subquestion. 

### 14 Onto number 3 -- contribution to pollinator 

### 15 biodiversity. The question really is asking about the 

### 16 assessment endpoints and whether or not these endpoints address 

### 17 a protection goal. However, we feel that the protection goal is 

### 18 very broad. It is sort of like saying we're going to cure 

### 19 cancer, or we are going to save the world - we are going to stop 

### 20 global warming. It's just a very, very large goal, and 

### 21 likewise, the assessment endpoint is very, very broad to species 

### 22 richness and abundance. Furthermore throughout the White Paper, 

### 23 there really are no measures for species richness and abundance 

### 24 taken. 

### 25 So we recommend that the protection goal be 

### 26 rewritten to read contribution to bee biodiversity. If you are

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1 talking about pollinators, this would mean all pollinators, 

### 2 insect pollinators and mammals and other vertebrates like 

### 3 humming birds and so on. It just seems beyond the scope of this 

### 4 White Paper. 

### 5 The honey bees - the entire genus Apis - they 

### 6 are not native to the Americas, and there is approximately 4000 

### 7 species of bees that are native to the Americas. New species 

### 8 are discovered every year. Just in our laboratory alone, we 

### 9 probably discover two every year. This year, there was a new 

### 10 species discovered in the middle of New York City. So, that's 

### 11 how poorly they're known. I mean, there are 4000 species that 

### 12 are known, but - so, we just wanted you to keep that in mind as 

### 13 you think about this goal. I think that is the end of what I 

### 14 have for (a). It is hard for me to separate out (a), (b) and 

### 15 (c), but we will get to (b) and (c). 

### 16 **DR. DANIEL SCHLENK:**Okay. Our first 

### 17 associate discussant is Dr. Berenbaum. 

### 18 **DR. MAY BERENBAUM:**I concur. 

### 19 **DR. DANIEL SCHLENK:**And Dr. Pettis. 

### 20 **DR. JEFF PETTIS:**I concur as well. 

### 21 **DR. DANIEL SCHLENK:**Dr. Pistorius? 

### 22 **MR. JENS PISTORIUS:**I concur as well. 

### 23 **DR. DANIEL SCHLENK:**Dr. Ostiguy. 

### 24 **DR. NANCY OSTIGUY:**I concur also. 

### 25 **DR. DANIEL SCHLENK:**All right, so let's open 

### 26 it up to other Panel members for any other comments for Question

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1 1(a). Anything else to add? Okay. Let's - who's going to be - 

### 2 any questions for clarification for the Agency for that? We're 

### 3 good? All right, thanks. All right, you want to read in 1(b), 

### 4 Mr. Moriarty. 

### 5 **MR. THOMAS MORIARTY:**Please comment on 

### 6 whether the measurement endpoints at the level of the colony, 

### 7 for examples, colony strength and survival, contamination of 

### 8 pollen and nectar, and species richness and abundance identified 

### 9 in Table 1 are consistent with the assessment endpoints 

### 10 identified in the Table and any additional assessment endpoints 

### 11 discussed in Part (a) of this question. Please include a 

### 12 discussion of any additional measurement endpoints that may be 

### 13 necessary to represent those assessment endpoints. 

### 14 **DR. DANIEL SCHLENK:**And again, Dr. James, you 

### 15 are our lead discussant for that question. 

### 16 **DR. ROSALIND JAMES:**All right, this is 

### 17 Rosalind James. So, again we are still working off of Table 1 

### 18 and now we are on the third column measurement of endpoints and 

### 19 how these meet the protection goals. So, back again to bee 

### 20 diversity, 98 percent, probably closer to 99 percent of bee 

### 21 species are solitary, and thus they do not have colonies. And, 

### 22 so a measurement of the endpoint colony strength would not 

### 23 really be measuring these bees directly. We want to make sure 

### 24 that that's clear. However, if honey bees serve as a surrogate 

### 25 for all bees, then this endpoint should be fine, but EPA needs 

### 26 to keep in mind that this is honey bee serving as a surrogate,

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1 so it is addressing honey bees and other bees. We also wanted 

### 2 to add to this colony strength and survival is listed, but we 

### 3 also wanted to add colony development, which means the growth of 

### 4 the colony during growth season. 

### 5 For the second item, production of hive and 

### 6 hive products, again, we recommend removing the word quality and 

### 7 also we wanted to add other hive products to this list. In 

### 8 particular, pollen, propolis and royal jelly; so that this 

### 9 element would now read, "quantity of hive products; and residues 

### 10 levels in honey, pollen, wax, propolis and royal jelly." 

### 11 And this was partly keeping in mind for honey 

### 12 bees and then for the native bees as well. Some of these plant 

### 13 resins and so on are very important to native bees in their 

### 14 natural habitat. If they are contaminated with pesticides, 

### 15 especially the larvae would be exposed. 

### 16 So then onto number 3 - contribution to bee 

### 17 biodiversity. What's currently the measurement endpoints are 

### 18 colony, strength and survival, and the second element is species 

### 19 richness and abundance. Colony survival is repeated here from 

### 20 up above and that's fine, but all my earlier comments about 

### 21 colony strength and survival apply here as well, and especially 

### 22 now we are talking about not honey bees, but other bees. So, 

### 23 those comments are doubly important here. 

### 24 Then species richness and abundance - again, 

### 25 this is a noble goal, but it's not really included anywhere in 

### 26 the assessments that occur in the White Paper. There's no way

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1 that you can assess species diversity using only one species, 

### 2 the honey bee. Again, this is a species in addition, 

### 3 essentially a domestic animal that's not native to any of the 

### 4 Americas. So, species richness and abundance is - it would be a 

### 5 good measurement endpoint, but you haven't addressed it at all. 

### 6 All right, that is the end of my comment. 

### 7 **DR. DANIEL SCHLENK:**Thank you. Dr. 

### 8 Berenbaum. 

### 9 **DR. MAY BERENBAUM:**I just want to clarify or 

### 10 emphasize that taking quality out of the measurement endpoint 

### 11 for hive products does not affect the validity of residue 

### 12 levels. So, quality independent of residue levels is not really 

### 13 the issue, but residue levels certainly do affect quality. 

### 14 **DR. DANIEL SCHLENK:**Okay. Dr. Pettis? 

### 15 **DR. JEFF PETTIS:**I concur with what's been 

### 16 said. 

### 17 **DR. DANIEL SCHLENK:**And Dr. Pistorius? 

### 18 **MR. JENS PISTORIUS:**Same here. 

### 19 **DR. DANIEL SCHLENK:**Dr. Ostiguy? 

### 20 **DR. NANCY OSTIGUY:**I just wanted to add that 

### 21 in looking at the using of honey bees to measure biodiversity, 

### 22 one of the things that we discussed are the studies that have 

### 23 been published on the negative impact of the presence of honey 

### 24 bees on native species. So, it's a significant consideration 

### 25 that we not use honey bees as a measurement of biodiversity. 

### 26 **DR. DANIEL SCHLENK:**Okay. Yes, Dr. James?

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1 **DR. ROSALIND JAMES:**This is Rosalind James. 

### 2 I want to add to that. At the foot note of this Table, and I 

### 3 believe it was said elsewhere in the White Paper also, the very 

### 4 last sentence says, in addition, protection of the honey bee 

### 5 would contribute to pollinator diversity indirectly by 

### 6 preserving the pollination and propagation of many plant species 

### 7 pollinated by honey bees, which also serve as food sources for 

### 8 other pollinating insects. Although, we have not been asked to 

### 9 edit this document, I would recommend deleting that because of 

### 10 the issue that Nancy brought up that many people - there's 

### 11 considerable body of research investigating how honey bees 

### 12 actually compete with native bees. So honey bees in themselves 

### 13 are not a good measure of bee diversity. 

### 14 **DR. DANIEL SCHLENK:**Dr. Fefferman? 

### 15 **DR. NINA FEFFERMAN:**Hi, I'm not a discussant 

### 16 on this question, so I hope this is still okay. I wanted to 

### 17 mention that maybe instead of looking just at measurements of 

### 18 biodiversity also to consider possibly as a measurement, the 

### 19 genetic variation seen within a species; because long-term 

### 20 conservation, especially population genetics models tell us that 

### 21 when we look for effective population sizes, those can be quite 

### 22 important. So, if we are looking at end measures of quality in 

### 23 the long term instead of the short term, along the same lines of 

### 24 biodiversity, genetic variation might be useful. 

### 25 **DR. DANIEL SCHLENK:**Thank you. Any other 

### 26 comments from panel members? Okay. Dr. James, I am sure that

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1 you are going to get Dr. Fefferman's comments into that? Okay. 

### 2 Great. All right. Let's go to the Agency. Any questions of 

### 3 clarification? Okay. Great. Moving right along. Okay. Mr. 

### 4 Moriarty, you want to read in 1(c) please? 

### 5 **MR. THOMAS MORIARTY:**Thank you. Question 1, 

### 6 subpart (c) - please comment on whether the measurement 

### 7 endpoints at the level of the individual bee, individual adult 

### 8 and larval survival, queen fecundity, brood emergence success 

### 9 and worker longevity, identified in Table 1 are consistent with 

### 10 the assessment endpoint identified in the Table and any 

### 11 additional assessment endpoints discussed in part (a) of this 

### 12 question. Please include a discussion of any additional 

### 13 measurement endpoints that may be necessary to represent those 

### 14 assessment endpoints. 

### 15 **DR. DANIEL SCHLENK:**And Dr. James, lead 

### 16 discussant. 

### 17 **DR. ROSALIND JAMES:**All right, so we are 

### 18 still on Table 1 and now looking at the fourth column 

### 19 measurement endpoints at the individual level and how this meets 

### 20 the protection goals. The items listed as measurement endpoints 

### 21 are very similar among the three different goals, and therefore 

### 22 I am going to discuss them all together. 

### 23 Individual worker survival is a very good 

### 24 measure. Larval survival should also be included. It's not 

### 25 listed here. Larval bioassays, I think, are going to be a very, 

### 26 very important contribution to understanding the possible

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1 effects of pesticides on bees. 

### 2 We suggest here using the word larvae and not 

### 3 brood, so that it's clearly inclusive of more than just honey 

### 4 bees. Brood is a word that is used basically for different 

### 5 larval stages and pupae of honey bees, even though the test will 

### 6 be for honey bees. 

### 7 We also recommend adding larval development 

### 8 time as one of your measurement endpoints; in particular, the 

### 9 time from egg lay to when a cell is capped. The negative 

### 10 effects of some pesticides such as insect growth and regulators 

### 11 may be able to be best detected using this sublethal measure. 

### 12 Also, we were not clear what is meant by brood 

### 13 success, which is on the products of the hive, so it's on the 

### 14 second box. We recommend changing this to larval survival and 

### 15 delayed development instead of brood success, which is 

### 16 essentially a repeat of what I just said above for the previous 

### 17 box. 

### 18 Brood size is a good measure, and there are 

### 19 fairly standard ways for measuring this in a hive. However, 

### 20 some people might think that this means individual larval size. 

### 21 So we recommend changing the wording to brood nest size and 

### 22 define it as the area of the sealed brood in a colony. 

### 23 Queen fecundity is a very useful endpoint, but 

### 24 at this time, it's difficult to measure. Perhaps other people 

### 25 on the Panel would want to make some comments on fecundity and 

### 26 the utility of using this and how it might be measured. That's

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1 the end of my comments. 

### 2 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 3 **DR. MAY BERENBAUM:**One of the parameters that 

### 4 are difficult to quantify is worker bee longevity because it 

### 5 tends to vary over the life cycle of the bee. The assays that 

### 6 exist remove the bee from its social context, so their meaning 

### 7 of the results is difficult to ascertain. So, in terms of 

### 8 reliable endpoints, worker bee longevity may present some 

### 9 experimental challenges. 

### 10 **DR. DANIEL SCHLENK:**Okay. Dr. Pettis you're 

### 11 next. 

### 12 **DR. JEFF PETTIS:**Just a couple of comments. 

### 13 One is for protection goal three and the measurement endpoints. 

### 14 Since it is a biodiversity or bee biodiversity, we are 

### 15 recommending that it be bee biodiversity protection goal. Then 

### 16 brood success for solitary pollinators would be a very excellent 

### 17 measure if you were testing solitary bees; the female's brood 

### 18 production would be an excellent measure and endpoint. 

### 19 Then last, the queen fecundity issue, I see 

### 20 that as a research gap, that we don't have adequate ways to 

### 21 measure queen fecundity at this time. 

### 22 **DR. DANIEL SCHLENK:**Thank you. Dr. 

### 23 Pistorius? 

### 24 **MR. JENS PISTORIUS:**Adding to the queen 

### 25 fecundity, I think we have a possibility of an indirect 

### 26 measurement by the assessment of brood and colony development.

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1 That is the only method available. 

### 2 **DR. DANIEL SCHLENK:**Okay. And Dr. Ostiguy? 

### 3 **DR. NANCY OSTIGUY:**I concur with the 

### 4 comments. 

### 5 **DR. DANIEL SCHLENK:**Okay. Any other panel 

### 6 members want to chime in on this one? Yes, Dr. Hunt. 

### 7 **DR. GREG HUNT:**I am just responding to what 

### 8 May Berenbaum said. Worker longevity does present some 

### 9 problems, but if you could measure it as kind of an average 

### 10 worker longevity if you, for example, mark the bees and then 

### 11 take photographs of the comb, so that could be a possibility. 

### 12 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 13 **DR. MAY BERENBAUM:**Yeah, just to stay it will 

### 14 present experimental challenges. That value is still going to 

### 15 vary depending on season and timing and all kinds of other 

### 16 factors. I'm not saying not to use it, but it can be hard to 

### 17 standardize. 

### 18 **DR. DANIEL SCHLENK:**Yes, Dr. Tarpy? 

### 19 **DR. DAVID TARPY:**This is Dave Tarpy. I would 

### 20 probably feel a little more comfortable rather than using kind 

### 21 of a more restrictive term of queen fecundity in this context to 

### 22 really mean something more like queen reproductive potential, 

### 23 which incorporates other things. Fecundity, at least to me - I 

### 24 could be a minority of one here - but fecundity is really kind 

### 25 of egg laying rate. I think there are other factors dealing 

### 26 with queen reproductive success and reproductive potential that

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1 could be important and captured here. 

### 2 **DR. DANIEL SCHLENK:**I see a lot of heads 

### 3 nodding, so I assume that's a consensus sort of statement there. 

### 4 Okay. Any other comments on 1 (c)? 

### 5 Okay. Hearing none let me go back to the 

### 6 Agency. Do you have what you need for that particular question? 

### 7 Fabulous. Let me just say thanks to Dr. James and that group 

### 8 for getting together and sort of expediting things. I really 

### 9 appreciate that. Okay. Move on to charge question 2. Again, 

### 10 if you wouldn't mind reading the subletters for 2, that would be 

### 11 great. 

### 12 **MR. KEITH SAPPINGTON:**Keith Sappington, EPA. 

### 13 Charge question 2, subpart (a) - please comment on whether the 

### 14 conceptual models depicted in Figures 4 through 8 are consistent 

### 15 with the protection goals and assessment endpoints identified in 

### 16 Table 1 and as discussed in question 1. 

### 17 **DR. DANIEL SCHLENK:**Okay. And lead 

### 18 discussant for this particular question is Dr. Ostiguy. 

### 19 **DR. NANCY OSTIGUY:**First comment - the 

### 20 conceptual models are very good. Lots of detail. We have 

### 21 comments that will help, I hope, fine tune things. I am going 

### 22 to start with some general comments, and then I have specific 

### 23 comments on specific figures. 

### 24 At least according to our reading at this 

### 25 point, none of the models include wax contamination, most 

### 26 specifically for larvae. They are sitting in contact with wax

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1 during the larval stage. Wax is included, sort of after the 

### 2 hive worker, the nurse bee, but it needs to also be looked at as 

### 3 an exposure media for the immatures. 

### 4 One of the things that came up yesterday was 

### 5 the comment about adding the RQs for oral and contact for the 

### 6 larva. And because of, you know, the specific lifestyle of the 

### 7 larva, that probably would be a good idea to combine that 

### 8 information rather than working just from the oral contact. 

### 9 One of the other comments, again dealing with 

### 10 the larval stage, most of our comments - a lot of the comments 

### 11 are on larva because we are still, of course, developing the 

### 12 proper protocols for evaluating them -- is to actually look at 

### 13 the total consumption of food during the larval stage rather 

### 14 than a per day consumption. It would then give you a subacute 

### 15 test, but a more realistic test since the quantities of food 

### 16 consumed over time change rather significantly and the types of 

### 17 food change. So, one value might actually be more useful. 

### 18 This is probably going to come up in a number 

### 19 of locations. Since I'm talking about larva, what will have to 

### 20 be chosen is a particular - well, it probably would be the day 

### 21 that capping occurs, is that you would actually use the weight 

### 22 rather than a larval measurement. So, a lot of the things that 

### 23 we have talked about are done as a quantity of exposure per bee. 

### 24 25 While this has been the way that the 

### 26 apiculture research community has reported exposures in the

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1 past, it's not really appropriate when we are going to be using 

### 2 honey bees as a surrogate because the other non-honey bees are 

### 3 certainly different in sizes. 

### 4 A more general comment, the feeling is that 

### 5 the assorted dotted lines should be changed to be solid lines. 

### 6 They should not be removed at this point as less important 

### 7 exposures or the receptors, the receiving end. 

### 8 One of the reasons for wanting to include more 

### 9 exposure media, more receptors, is that we are using honey bees 

### 10 as a surrogate and there are going to be exposures that are 

### 11 potentially important to non-honey bees that otherwise would not 

### 12 be captured. So for instance, leaf cutter bees - please correct 

### 13 me if I get this wrong - leaf cutter bees used for alfalfa 

### 14 pollination, at this point, we probably bring in as many or 

### 15 nearly as many leaf cutting bees to pollinate alfalfa, some 

### 16 coming from as far away as Canada. 

### 17 These bees are in contact with soil. So 

### 18 excluding soil when we are doing the evaluation is not a good 

### 19 idea. In addition, since we are going to be using honey bees as 

### 20 a surrogate for, let's say tube nesting, solitary bees, they use 

### 21 mud for their partitions. So, we should really include that 

### 22 soil exposure when we are looking at the honey bees. Generally, 

### 23 the group did agree that, you know, the matrices that you 

### 24 created are appropriate for both honey bees and other assorted 

### 25 native bees. 

### 26 Two specific comments - figure 6 - there

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1 should be a line going from drift abraded seed coating as a 

### 2 source to the exposure media residues in or on pollen, nectar, 

### 3 exudates and honey dew. Then figure 7 needs to include dust as 

### 4 an exposure media. Then, residues - it should read for the 

### 5 exposure media of pollen, nectar and exudates, it should be 

### 6 residues in or on since we are looking at soil as a possible, or 

### 7 dust as a media. Those are the comments that I have at this 

### 8 point. 

### 9 **DR. DANIEL SCHLENK:**Thank you. Dr. Hunt, you 

### 10 are our first associated discussant. 

### 11 **DR. GREG HUNT:**I concur. 

### 12 **DR. DANIEL SCHLENK:**Thanks. Dr. Pistorius? 

### 13 **MR. JENS PISTORIUS:**I concur, except one 

### 14 comment for figure 6. As you have said, there's one line from 

### 15 drift abraded seed coating to residues in pollen and nectar, we 

### 16 are talking about residues on planted seeds, but what we mean is 

### 17 on off crop lands. 

### 18 **DR. DANIEL SCHLENK:**Okay. 

### 19 **MR. JENS PISTORIUS:**Or in crop lands if there 

### 20 is no tillage. 

### 21 **DR. DANIEL SCHLENK:**Okay. Yes, Dr. Ostiguy? 

### 22 23 **DR. NANCY OSTIGUY:**Nancy Ostiguy. I did not 

### 24 understand and didn't hear what you wanted in there. 

### 25 **MR. JENS PISTORIUS:**Okay. With this line 

### 26 drift of abraded seed coating, as you said, we want the line

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1 drift to residues in pollen, nectar and exudates and honey dew; 

### 2 but the box above is talking about residues on planted seeds. 

### 3 But what we mean is not drift on the planted seeds nor the 

### 4 secreted seeds that we bring into the soil, but we want to look 

### 5 at residues in pollen and nectar off crop or if there are 

### 6 flowering weeds in the area which is sown. Does that clarify? 

### 7 **DR. DANIEL SCHLENK:**Do you have that 

### 8 information? Perhaps just a written component of your comments, 

### 9 if you could send that, that would probably be appropriate as 

### 10 well just to confirm. Okay. Dr. Potter? 

### 11 **DR. THOMAS POTTER:**I concur with everything. 

### 12 I just wanted to add one small detail, and this is in regard to 

### 13 the soil exposure. I believe that one thing we may want to 

### 14 consider is the treated soil as a source of dust via wind 

### 15 erosion and eroded particles from soil via wind can be strongly 

### 16 enriched if pesticides and active ingredients are implied to 

### 17 them. So, that might be an added twist in terms of the exposure 

### 18 issues that we may be addressing in the case of pesticides and 

### 19 soil. 

### 20 **DR. DANIEL SCHLENK:**Okay. Thanks. And Dr. 

### 21 Schwab? 

### 22 **DR. PAUL SCHWAB:**Yes, I only have one comment 

### 23 and that's on figure 6. With the seed based translocation, the 

### 24 box from the residue on planted seeds, this is -- just that term 

### 25 seed based translocations is a bit confusing to me in itself. 

### 26 I'm not really sure what that means, and it could be - I don't

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1 know if this means that the seed itself is somehow assimilating 

### 2 the pesticide directly from the dust or the coating I mean, but 

### 3 I think it would help to clarify if we could consider the seed 

### 4 coating as becoming part of the soil as well, at least some 

### 5 fraction of that, and then being subject to uptake trough the 

### 6 roots. Maybe that's what is implied already, but it wasn't 

### 7 clear to me. But otherwise, I concur with all the other 

### 8 statements. 

### 9 **DR. DANIEL SCHLENK:**Okay. Other Panel 

### 10 members? Yes, Dr. Berenbaum. 

### 11 **DR. MAY BERENBAUM:**Did the group discuss 

### 12 whether there is a significant possibility of exposure via 

### 13 contamination of resins used for propolis, among other things, 

### 14 populus as a biofuel crop tree and is a common source for bees 

### 15 to harvest propolis materials? 

### 16 **DR. DANIEL SCHLENK:**It would appear not. 

### 17 Okay. Dr. James and then Dr. Hunt. 

### 18 **DR. ROSALIND JAMES:**I've a few different 

### 19 comments - it's a little bit hard to separate out the different 

### 20 questions. I will be addressing some of this in the next 

### 21 question. I am probably the proponent here for the non-Apis 

### 22 bees. These diagrams are a good place where the non-Apis bees 

### 23 don't show up. Dr. Ostiguy already talked a little bit. I just 

### 24 wanted to clarify some of the comments she made. 

### 25 So, the leaf-cutter bees will be using leaf 

### 26 pieces and the larvae will be growing up in an environment with

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1 leaf pieces. So residues that are on these leaf pieces would 

### 2 have a different experience for alfalfa-leaf cutting bees than 

### 3 for honey bees. Honey bees would be that foraging adults would 

### 4 land on them as opposed to larvae being raised inside them. The 

### 5 majority of solitary bees are actually ground-nesting bees. 

### 6 So, we heard today with public comments and in 

### 7 some of the public comments, and again it shows up over and over 

### 8 in the diagrams that bees do not come into contact with soil. 

### 9 Well, the majority of the wild bees grow up in the soil. 

### 10 And there are lots of models, some of which 

### 11 are presented here about tracking what the soil residues might 

### 12 be and those could be applied, perhaps, to include in-contact 

### 13 toxicity test with larvae. So again, in-contact toxicity tests 

### 14 on larvae are often excluded because honey bee larvae would not 

### 15 be exposed to the soil, but many of the solitary bee larvae 

### 16 would. 

### 17 Also, I would like to reiterate a comment that 

### 18 was made yesterday in one of the questioning that what about 

### 19 nonsystemic soil applied pesticide. For the same reason the 

### 20 bees would actually be exposed to the soil, so even though it is 

### 21 not a systemic pesticide, it could have some affect on the 

### 22 non-Apis bees. 

### 23 Throughout the discussions yesterday, I was 

### 24 kind of struck by a very crop-centric view, but we are actually 

### 25 talking about an environmental impact and we have flowering 

### 26 plants in the crops that are not part of the crop. You can have

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1 overspray and so on, and the bees would be visiting some of 

### 2 these. 

### 3 I am not exactly sure where to put it into 

### 4 these diagrams, but certainly flowering weeds, or in my 

### 5 laboratory - I work for the Agricultural Research Service, and I 

### 6 know some of the other folks here are really trying to encourage 

### 7 farmers to intentionally plant flowering plants to increase the 

### 8 bloom period and the blood resources for bees. How do we 

### 9 include that in a pesticide assessment if there is overspray of 

### 10 pesticide? 

### 11 Pesticides are a very big problem for us in 

### 12 trying to introduce non-Apis bees into farming crop systems. 

### 13 These are some of the areas. That's it. 

### 14 **DR. DANIEL SCHLENK:**Thanks. Dr. Hunt? 

### 15 **DR. GREG HUNT:**I don't know if -- this 

### 16 comment may not be as important, but in talking about what May 

### 17 Berenbaum was saying about propolis, this is collected from 

### 18 typically trees that are not directly exposed to pesticide, but 

### 19 propolis in the hive could absorb pesticides and residues could 

### 20 be there. 

### 21 **DR. DANIEL SCHLENK:**Great. Dr. Ostiguy, I 

### 22 see you typing this up, so I'm assuming you are putting all this 

### 23 in. Great. Any other Panel members? Dr. Tarpy? 

### 24 **DR. DAVID TARPY:**This is Dave Tarpy. I've 

### 25 noticed on these flowcharts, which I agree are very good and 

### 26 they also are very kind of Apis centric and don't quite capture

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1 some of the subtleties of the solitary bees. But with that 

### 2 said, the main receptors in these flowcharts as they ought to be 

### 3 are the foraging bees. In doing so, the arrows that point to 

### 4 them really, it's implying that the vast majority of the 

### 5 ingestion is with the foraging bees. 

### 6 I know that the presentations really focus on 

### 7 that with nectar and is true, but really they are transporting, 

### 8 but not always ingesting, especially when it comes to pollen. 

### 9 So, there are some subtleties where hive bees, for example -- 

### 10 the nurses are kind of the direct consumers, the ones that 

### 11 ingest pollen, for example, rather than the nurse bees. So, I 

### 12 don't know if that really affects the overall concept of these, 

### 13 but there is a division of labor when it comes to ingestion. It 

### 14 doesn't all go through the foragers per se. 

### 15 **DR. DANIEL SCHLENK:**Okay. Thank you. Any 

### 16 other comments from the Panel members? Okay. Let me go to the 

### 17 Agency - any questions or clarification? 

### 18 **MR. KEITH SAPPINGTON:**Keith Sappington. I 

### 19 just have clarifying question on the figures. Do you want us to 

### 20 move the wax and propolis, which are found in the figures to a 

### 21 different location? 

### 22 **DR. DANIEL SCHLENK:**Who wants to address 

### 23 that? Dr. Ostiguy, do you want to try to address that? 

### 24 **DR. NANCY OSTIGUY:**Yes, I will try - this is 

### 25 Nancy Ostiguy. Yes, because it is an exposure media for larvae 

### 26 whereas where it is right now is a receptor, at least the way

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1 that I'm reading it, wax seems to be incidental? 

### 2 **MR. KEITH SAPPINGTON:**Keith Sappington. Yes, 

### 3 in terms, I think I understand the comment now that along the 

### 4 boxes that are listed as receptors, we have receptors, but we 

### 5 also have essentially exposure media that are relevant to those 

### 6 receptors, and wax and propolis is listed as an exposure media 

### 7 for the brood, for the larvae. But that is not necessarily 

### 8 tracking with the bold - is that the question that that's not 

### 9 tracking with where the exposure media are listed above it? 

### 10 **DR. DANIEL SCHLENK:**Dr. James? 

### 11 **DR. ROSALIND JAMES:**Yeah, I would like to 

### 12 address that. I agree with what you suggested actually that 

### 13 moving the honey wax and propolis as an exposure media instead 

### 14 of being an attribute change - I mean, it may also be an 

### 15 attribute change, but that's more with human. Here? I think he 

### 16 meant here. Oh, we don't know what you mean. Is it here or 

### 17 here? At receptors or attribute change? You have it in two 

### 18 places. 

### 19 **DR. NANCY OSTIGUY:**This is Nancy Ostiguy. I 

### 20 think that we were talking about putting wax and propolis as an 

### 21 exposure media separate from and maybe in addition to where you 

### 22 have it right now in the receptors. 

### 23 **MR. KEITH SAPPINGTON:**Okay. I can 

### 24 understand. Thank you. 

### 25 **DR. DANIEL SCHLENK:**Okay. Everybody clear on 

### 26 that? Oh, Dr. James?

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1 **DR. ROSALIND JAMES:**I'm the slow one, in the 

### 2 head here, I guess. To me, you don't really have it as a 

### 3 receptor. You have it on top of the arrow, what does that mean? 

### 4 5 **MR. KEITH SAPPINGTON:**Yes, on top of the 

### 6 arrow means a mechanism by which, in this case it would be the 

### 7 bee brood and the queen would be exposed, so they would be 

### 8 exposed via brood provisions, contact with wax, propolis and 

### 9 royal jelly. 

### 10 So, it was just to rather than have a blank 

### 11 arrow going to them from the hive bees, nurse, worker, drones, 

### 12 which all participate in processing that material, it was just 

### 13 to illustrate how they would be exposed. But it could easily be 

### 14 put above under exposure media and avoid that confusion. 

### 15 **DR. ROSALIND JAMES:**This is Rosalind James. 

### 16 It could be in both. I mean, because it really is then an 

### 17 exposure media even by your explanation and by putting it on the 

### 18 arrow, it just sort of explains how it is an exposure media. 

### 19 **DR. DANIEL SCHLENK:**Dr. Ostiguy? 

### 20 **DR. NANCY OSTIGUY:**This is Nancy Ostiguy. 

### 21 One of the reasons in addition to placing wax and propolis as an 

### 22 exposure media is that those particular materials are recycled 

### 23 by the workers. So, they're going to then repeatedly be taking 

### 24 it up and placing it back down again. 

### 25 **DR. DANIEL SCHLENK:**Dr. Hunt? 

### 26 **DR. GREG HUNT:**I hate to complicate things,

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1 but in the exposure media could also have an arrow down to the 

### 2 hive bees, all the adults, because they are exposed by the wax. 

### 3 Their exoskeleton dissolves liquids. 

### 4 **DR. DANIEL SCHLENK:**Yes, Dr. Ostiguy? And 

### 5 then Dr. Berenbaum? 

### 6 **DR. MAY BERENBAUM:**And to complicate things 

### 7 further; the royal jelly arrow goes from hive bees to queens, 

### 8 but in fact larvae eat royal jelly too. So, all three of those, 

### 9 brood provisions, wax propolis, royal jelly, should go to that 

### 10 joint line because they go to both queens and to regular worker 

### 11 grubs. 

### 12 **DR. DANIEL SCHLENK:**Dr. Ostiguy? 

### 13 **DR. NANCY OSTIGUY:**This is Nancy Ostiguy. I 

### 14 think we've talked a little bit about that yesterday where there 

### 15 was sort of an - because the conceptual model just would get 

### 16 incredibly messy if we had lines all over the place, which might 

### 17 be the argue for having provisions, wax, propolis, royal jelly 

### 18 up in the exposure media maybe to clarify that everything is 

### 19 moving. 

### 20 **DR. DANIEL SCHLENK:**Okay. Dr. James? 

### 21 **DR. ROSALIND JAMES:**I'm hoping to simplify, 

### 22 not complicate here. So, maybe your bee brood and your queen 

### 23 boxes could be one box called larvae, queen and worker, drone 

### 24 larvae, all of them. 

### 25 **DR. DANIEL SCHLENK:**Okay. Let me go back to 

### 26 Keith. It sounds like you have a few options here.

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1 **MR. KEITH SAPPINGTON:**I think we understand 

### 2 some of the confusion. Should the Panel have time or feel it 

### 3 useful to actually suggest the modifications in the report or 

### 4 have a revised figure in the report that might also clarify 

### 5 things. And we could even provide the electronic versions if 

### 6 necessary because it takes time to. 

### 7 **DR. NANCY OSTIGUY:**This is Nancy Ostiguy. 

### 8 That would be wonderful, and then we can make sure that we are 

### 9 clear on what we are trying to tell you. 

### 10 **DR. DANIEL SCHLENK:**If you would email that 

### 11 to Fred that would be great and then he can distribute that. 

### 12 Other than that, everything is clear? Clear as honey? Sorry. 

### 13 I was going to say mud, but - all right. 

### 14 We are ready to move onto question three. Who 

### 15 is going to be reading that in? Okay. Mr. Moriarty? Again, 

### 16 please read (a) if you wouldn't mind. 

### 17 **MR. THOMAS MORIARTY:**Question 3, subpart (a) 

### 18 - please comment on the extent to which the assessment of risk 

### 19 to the honey bee may serve to meet the protection goals 

### 20 identified in the White Paper, that is protection of pollination 

### 21 services, protection of honey and hive product production, and 

### 22 protection of pollinator diversity. 

### 23 **DR. DANIEL SCHLENK:**Okay. Dr. James, you are 

### 24 the discussant. 

### 25 **DR. ROSALIND JAMES:**This is Rosalind James. 

### 26 I want to make a request. I want to address (a) and (b)

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1 together. They are hard for me to separate. Can I do that? 

### 2 **DR. DANIEL SCHLENK:**Sure. 

### 3 **DR. ROSALIND JAMES:**Do you want to read (b) 

### 4 also? 

### 5 **Dr. DANIEL SCHLENK:**Do you want to read (b) 

### 6 in also then if we are going to do that? 

### 7 **MR. THOMAS MORIARTY:**3(b) - until guidelines 

### 8 are developed for testing non-Apis species of bees, please 

### 9 comment on the extent to which the honey bee may or may not 

### 10 serve as a reasonable surrogate for non-Apis bees. Given the 

### 11 differences in lift history, strategies, and potential different 

### 12 sensitivities to pesticide toxicants, please include a 

### 13 discussion of which types of non-Apis bees may be particularly 

### 14 well represented by either the individual level or the colony 

### 15 level endpoints identified in Table 1 of the White Paper, as 

### 16 well as which types of non-Apis bees may not be well represented 

### 17 and therefore may be the focus of potential research areas in 

### 18 the future. 

### 19 **DR. DANIEL SCHLENK:**Okay. Dr. James? 

### 20 **DR. ROSALIND JAMES:**This is Dr. James. To me 

### 21 these two questions kind of go together. Due to the large 

### 22 diversity of non-Apis bees, I understand the EPA cannot test a 

### 23 significant number of them and furthermore, most of them are 

### 24 wild and they are not in production in any way and this makes it 

### 25 difficult to do Tier I types of testing. 

### 26 So, I think that honey bees probably could

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1 make a good surrogate species to help represent the 

### 2 susceptibility of other bees, but I would implore EPA to 

### 3 recognize that it is a surrogate species that represents other 

### 4 bees. This then affects how you - this part addresses (a), in 

### 5 which how you do the risk assessment. So, when you come up with 

### 6 your flow diagrams and when you come up with your measures of 

### 7 exposure and so on, you have to keep these other bees in mind as 

### 8 well as a honey bee. 

### 9 The biology of these other bees is diverse and 

### 10 great and it's hard to know about all of the possibilities, but 

### 11 I think there are some basic principles that you could learn. 

### 12 We discussed some of them already, exposure to leaf pieces of 

### 13 the larvae and nesting in the ground. Another one is the 

### 14 seasons in which they fly are going to be very different than 

### 15 for honey bees. 

### 16 But there are some things that also have to be 

### 17 kept in mind where honey bees may not be very good surrogates 

### 18 for representing these other bees. So, as a stop gap measure, 

### 19 maybe honey bees are adequate, but there has been discussion 

### 20 within the group of whether really you ought to include some 

### 21 other solitary bee to help keep in mind some of the particulars 

### 22 that occur with solitary bees. 

### 23 In particular, we discussed that honey bees, 

### 24 because of their colony behavior and the fact that the colony 

### 25 lives for multiple years, they store food and they have large 

### 26 number of workers and they are able to change status of the

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1 workers from being nurse bees to foragers and so on, to 

### 2 compensate for various environmental factors. So, they can ride 

### 3 through essentially bad times or they can withstand a certain 

### 4 amount of mortality among the workers. 

### 5 But with solitary bees or something like most 

### 6 of the social non-Apis bees that have very small colonies or 

### 7 their colonies are annual, they don't have the ability to 

### 8 withstand these kinds of stresses. An example would be with 

### 9 pollen storage. 

### 10 With your solitary bees and with bumble bees 

### 11 too, the bees go out and collect pollen - I will back up. With 

### 12 solitary bees, the female provisions each one of her larvae. So 

### 13 the female is going to go out and she is going to have one whole 

### 14 provision made for that larva at a particular time. There may 

### 15 be a pesticide exposure out of that time. The entire life of 

### 16 that larva is going to be exposed to that pesticide. A later 

### 17 larva may not have any exposure to pesticide if it's a 

### 18 short-lived pesticide out in the field. 

### 19 Whereas with honey bees, maybe they could 

### 20 avoid the pesticide and live on reserves within the colony if 

### 21 there is an avoidance mechanism - so their exposure would be not 

### 22 as forced as it is with the solitary bees. 

### 23 We also had a considerable amount of 

### 24 discussion about the pollen and how much pollen the honey bees 

### 25 consume. I think over lunch time, we resolved that. Your 

### 26 numbers are probably not that inaccurate, they are probably a

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1 good estimate. What threw us off was your use of nectar instead 

### 2 of honey. If we take your nectar numbers and divide them by 

### 3 three, then we come up with a more reasonable estimate of honey 

### 4 consumption. 

### 5 With your solitary bees and your bumble bees, 

### 6 to some extent they don't consume as much honey. They don't 

### 7 consume any honey - bumble bees make some honey. It's not as 

### 8 processed as honey bee honey. Your solitary bees generally 

### 9 don't have honey. They just collect nectar and the primary food 

### 10 source is really going to be pollen. So, probably more pollen 

### 11 based. 

### 12 When I look at numbers from my laboratory, I 

### 13 have numbers from this summer and I know what size my pollen 

### 14 provisions are. I divide them by 14 days, that's how long the 

### 15 larvae take to live. I come up with pollen consumption rates 

### 16 that are similar to what you're finding for honey bees. These 

### 17 bees are much smaller, but they are eating about the same amount 

### 18 of pollen per day. 

### 19 So, another bee such as the Osmia lignaria, 

### 20 the blue orchard bee, which has a size more comparable to honey 

### 21 bees, are probably going to eat more pollen than honey bees. 

### 22 So, there is probably more pollen consumption in the non-Apis 

### 23 bees; and certainly the ratio of pollen to nectar is going to 

### 24 probably be higher than it is with honey bees. 

### 25 Again - I don't want to reiterate too much, 

### 26 but I'm kind of bent out of shape a little bit about not doing

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1 contact toxicity to solar soil-applied pesticides - I think that 

### 2 should be included. 

### 3 For the honey bee test, even though honey bees 

### 4 will never be exposed probably to soil pesticides, other bees 

### 5 are likely to be exposed to soil pesticides. So, there should 

### 6 be some contact toxicity done for those. 

### 7 The other big difference is some of your 

### 8 solitary bees are going to be more specialists than honey bees. 

### 9 The honey bees are what we call generalist bee. They are 

### 10 probably the extreme of generalist bees. Bumble bees also are 

### 11 fairly general. They will forage on most any kind of flowering 

### 12 plant. Not any kind of flowering plant, but they have a very 

### 13 broad host range. Your solitary bees tend to have a narrower 

### 14 host range. 

### 15 So with the mason bees, the Osmia lignaria, 

### 16 they tend to forage mainly on fruit trees or flowering trees. 

### 17 They are not going after only one plant, but there's a much 

### 18 smaller group of plants that they go after. This will be true 

### 19 for many of your native bees. So, they are going to be very 

### 20 specialized. Somehow, this should be taken into consideration 

### 21 in the risk - what do you call -- the risk management aspect I 

### 22 think, if I understand the difference between risk assessment 

### 23 and risk management. 

### 24 So when you are depending on the pesticide and 

### 25 where it's being applied, this may affect some bees more than 

### 26 others. So, perhaps the honey bee could be the surrogate in

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1 your Tier I bioassay, but in your risk assessment you need to 

### 2 include what specialist bees might be exposed under a particular 

### 3 circumstance. Especially - I mean not all applications are 

### 4 agricultural. So, we spray for mosquitos, we spray for forest 

### 5 pests and so on. 

### 6 Again, if you were to include one other bee, I 

### 7 thought about this question. Probably the easiest bee to 

### 8 include in addition to honey bee, and I don't think it's just a 

### 9 personal bias on my part, but would be Megachile rotundata, 

### 10 which is the alfalfa-leaf cutting bee. You can buy it by the 

### 11 gallon. 

### 12 Doing bioassays with them really is no more 

### 13 difficult than honey bees and probably easier, but they aren't 

### 14 honey bees, so you can't treat them exactly the same as honey 

### 15 bees. Certainly adult bioassays would be easy because you can 

### 16 buy the bees by the gallon, incubate them and emerge them as 

### 17 adults. 

### 18 Doing larval tests will be more difficult 

### 19 because like honey bee, you will have to collect larvae out in 

### 20 the field and that can only be done during the time that the 

### 21 bees are pollinating alfalfa. So, that's more difficult, but 

### 22 you could include adult alfalfa-leaf cutting bee or even blue 

### 23 orchard bee. You can buy those, but they are a little more 

### 24 expensive and they are not by the gallon, they are by the bee. 

### 25 So, they are about 50 cents a bee. 

### 26 At least - you know one thing, keep that in

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1 mind I guess as you go through the assessment. Something to 

### 2 force you to keep in mind, not the particularly of only honey 

### 3 bees if you want to use honey bees as a surrogate. 

### 4 **DR. DANIEL SCHLENK:**Okay. Thanks Dr. James. 

### 5 Dr. Berenbaum? 

### 6 **DR. MAY BERENBAUM:**I concur. 

### 7 **DR. DANIEL SCHLENK:**And Dr. Pettis? 

### 8 **DR. JEFF PETTIS:**I just have to add a bit to 

### 9 the using honey bees as a surrogate. It is somewhere along the 

### 10 lines of Dr. James's comment. Just that, I think two genera, 

### 11 Osmia and Megachile, both would make excellent things for Tier I 

### 12 testing. They can be used; you can do direct toxicity testing 

### 13 on adults and get some idea about the degree to which honey bees 

### 14 can serve because protection goal 3 is about bee biodiversity. 

### 15 So really to get at that, some testing using at least one or 

### 16 both of those species might be valid. 

### 17 **DR. DANIEL SCHLENK:**And Mr. Pistorius. 

### 18 **MR. JENS PISTORIUS:**And I would like to add 

### 19 those are probably also the easiest species to go to a higher 

### 20 tier tests, you want semi-field test, you can also use them. 

### 21 **DR. DANIEL SCHLENK:**Okay. Other Panel 

### 22 members? Dr. Ostiguy? 

### 23 **DR. NANCY OSTIGUY:**This is Nancy Ostiguy. I 

### 24 would like to add that we are sort of tossing out non-honey bee 

### 25 species that are useful for testing. Bumble bees are actually 

### 26 quite good for Tier II, because you can actually confine them

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1 and they forage normally. 

### 2 **DR. DANIEL SCHLENK:**Mr. Pistorius? 

### 3 **MR. JENS PISTORIUS:**Except one exception, the 

### 4 brood development is very difficult to assess because there are 

### 5 multiple layers of brood and you have no way of assessing them. 

### 6 That is why it is easier with Megachile or Osmia. 

### 7 **DR. DANIEL SCHLENK:**Yes, Dr. James? 

### 8 **DR. ROSALIND JAMES:**Why would brood 

### 9 assessment be so difficult? I mean you can put them in small 

### 10 colonies that you can open up and look at them. 

### 11 **MR. JENS PISTORIUS:**Because you cannot 

### 12 estimate the exact number of larvae and you cannot watch the 

### 13 individual development of larvae. When they make all those 

### 14 piles, one across the other, you don't know what is happening. 

### 15 **DR. DANIEL SCHLENK:**That was Mr. Pistorius. 

### 16 Okay. Any other Panel input here? Okay. Let me go to the 

### 17 Agency in terms of questions or clarification. Got a lot of 

### 18 comments here. Okay. So that wraps up charge question 3. 

### 19 Moving right along. Let's go to charge 

### 20 question 4. If you could read letter (a) there would be great. 

### 21 **MS. KRIS GARBER:**This is Kris Garber. Charge 

### 22 question 4, subpart (a) - please comment on the strengths and 

### 23 limitations of the proposed approach for assessing contact 

### 24 exposures to honey bees in Tier I exposure assessments, i.e. 2.7 

### 25 micrograms a.i. per bee per 1 pound a.i. per acre, which is 

### 26 based on the honey bee specific maximum concentration reported

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1 by Koch and Weisser, 1997. 

### 2 **DR. DANIEL SCHLENK:**Okay. Our lead 

### 3 discussant for that is Dr. Schwab. 

### 4 **DR. PAUL SCHWAB:**Okay. Thank you. This is 

### 5 Paul Schwab. This is a very important consideration because 

### 6 foliar sprays can result in some of the very highest levels of 

### 7 residues other than the foliar spray diet, and thus the highest 

### 8 level of exposure to the foraging bees. 

### 9 Well, there is a lack of data for bee contact 

### 10 and general exposure to key pesticides, so some sort of 

### 11 surrogate is necessary. That was the motivation, I'm assuming, 

### 12 behind this study in the first place. 

### 13 The approach that's being used has a lot of 

### 14 merit for a lot of reasons, but it also some limitations. 

### 15 Clearly in the absence of any data at all, and we will get to 

### 16 this in part (b), then a model would be necessary, and we will 

### 17 talk about that in a moment. 

### 18 Some of the strengths of this approach would 

### 19 be that the surrogate tracer was not being used anywhere else in 

### 20 the region, so there would be - there were a lot of concerns in 

### 21 other studies about bees going outside the region of control and 

### 22 getting into pesticides that weren't part of the study. 

### 23 That wouldn't be the case with this particular 

### 24 tracer because it's unique. This would not be applied to crops 

### 25 under any circumstances. And so, these compounding factors are 

### 26 eliminated. The concentration reported was the highest value

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1 that was normalized and that should be fairly protective. 

### 2 Some of the weaknesses would be that it is a 

### 3 bit of an old study - a 15-year-old study. It doesn't mean that 

### 4 it has no value, but it's possible that some updating could be 

### 5 in order. 

### 6 The compound that was used was a sodium 

### 7 fluorescein. It's a fluorescent dye. It's not really one of 

### 8 the pesticides we look at obviously. It may not even have many 

### 9 of the same properties that these pesticides would have, but 

### 10 nevertheless, it probably is a reasonable start for a surrogate 

### 11 and under these circumstances. 

### 12 Bees were actually foraging during the 

### 13 application and subject to direct spray, so this could result in 

### 14 some high estimates. This was not what was being studied, so 

### 15 any bees that were intercepting some of this spray would be 

### 16 subject to having anomalous concentrations, anomalously high. 

### 17 And then finally, the normalization -- and 

### 18 this is not just in this particular study, but elsewhere as well 

### 19 - but the normalization assumes the linear relationship between 

### 20 the rate of spraying and the rate of accumulation by bees, which 

### 21 is not necessarily valid. I will talk about this later in case 

### 22 there are some questions about this. Anyway, those are my 

### 23 comments. I know that Tom - you also have something? 

### 24 **DR. DANIEL SCHLENK:**Thanks. Dr. Potter? 

### 25 **DR. THOMAS POTTER:**Okay. I certainly concur 

### 26 with Dr. Schwab. I believe that this study was reasonable and

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1 certainly an elegant piece of work, and gets at some very 

### 2 important questions using a very novel approach. 

### 3 My primary concern about the study is in the 

### 4 manner of which the dye was applied to the leaves. It certainly 

### 5 does not necessarily - and its certainly unlikely to mimic how 

### 6 pesticides are applied or co-applied with a number of different 

### 7 adjuvants that are blended into the material, including 

### 8 spreaders and stickers and all sorts of other things. 

### 9 So, that makes this type of study, I guess 

### 10 somewhat questionable in terms of acting as a firm surrogate for 

### 11 pesticides and then until such time more work is done, I think 

### 12 there is considerable uncertainty there in terms of whether this 

### 13 accurately represents potential exposures or not. 

### 14 **DR. DANIEL SCHLENK:**Does anybody else want to 

### 15 chime in on this particular question? Okay. Dr. James? 

### 16 **DR. ROSALIND JAMES:**I don't know, Dave, we 

### 17 talked a little bit about by weight. I mean, and this is 

### 18 consumption, which the next question is exposure, but again as 

### 19 we move away from - as we move towards thinking of honey bees as 

### 20 a surrogate and not just as honey bees, bees come in different 

### 21 sizes and should the consumption rate be by weight or you have 

### 22 different -- drones are larger than workers and queens. Do you 

### 23 have anything you want to add to that? 

### 24 **DR. DAVID TARPY:**This is Dave Tarpy, NC 

### 25 State. Yeah, that was a comment and a question that I had for 

### 26 the EPA members yesterday about standardizing it to kind of per

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1 milligram of body weight of an individual rather than on a per 

### 2 bee basis. 

### 3 To kind of get to your point and to your 

### 4 question, question number 3 about using honey bees as a good 

### 5 surrogate, that would and I think - maybe I'm wrong here, I 

### 6 would like other comments on this -- but it seems to me that 

### 7 that would translate a lot more effectively to other life, to 

### 8 other castes and to other types of bees to do it on a per 

### 9 milligram basis. I was going to raise those points in question 

### 10 number 8 later on. But I think it's applicable here too. 

### 11 **DR. DANIEL SCHLENK:**I see a lot of heads 

### 12 nodding on that in terms of normalizing per mass it sounds like, 

### 13 in terms of per bee. Yeah, Dr. Pettis? 

### 14 **DR. JEFF PETTIS:**I agree with what Dr. Tarpy 

### 15 said. The one exception in honey bee colony would be the queen 

### 16 in which she, even by her body mass, she's actually processing a 

### 17 great deal more food than any other individual in the colony. 

### 18 She is kind of this egg laying machine, thus her nutritional 

### 19 requirements are extremely high relative to anybody else. So, 

### 20 body weight wouldn't account for that. 

### 21 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 22 **DR. MAY BERENBAUM:**Am I correct in assuming 

### 23 that Koch and Weisser's study involved only one race of bee, and 

### 24 again, there is race variation and size and pursuitness and 

### 25 other attributes. So, another argument for weight rather than 

### 26 per bee.

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1 **DR. DANIEL SCHLENK:**So again, let me just 

### 2 encourage you to get your written comments to Dr. Schwab at 

### 3 least for this particular question - oh sorry Dr. James, didn't 

### 4 see you over there. 

### 5 **DR. ROSALIND JAMES:**Sometimes it is hard to 

### 6 know where to put the comments, I guess; but also the 

### 7 consumption rate had been by day. The bee larvae have such a 

### 8 short life span, that it's often easier to measure it across 

### 9 their whole life span and with the honey bee, they are only 

### 10 eating for five days and mostly consumption is done in two days. 

### 11 There are some early. 

### 12 Some other possibility would not to be looking 

### 13 at the consumption per day, but over the lifecycle, I believe 

### 14 for the larvae. This also would be very applicable to the 

### 15 solitary bees where they have given them mass provision and then 

### 16 that is what they eat is that mass provision. Again as I said, 

### 17 their exposure - whatever day that was collected, that's the 

### 18 exposure level that they are stuck with unless, of course, you 

### 19 have degradation over time, but it's a relatively short period 

### 20 of time that they are feeding on. 

### 21 So, I did want to suggest not looking at 

### 22 consumption rate per day for larvae, but at total consumption 

### 23 for larvae and then the question again about the pollen versus 

### 24 the nectar, and if there is a difference in pesticide 

### 25 concentration between pollen and nectar, that ratio could be 

### 26 very important. Different life stages have different ratios.

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1 Your solitary bees are going to be more dependent on pollen than 

### 2 your worker honey bees as adults. Thank you. 

### 3 **DR. DANIEL SCHLENK:**Sure. Dr. Fefferman? 

### 4 **DR. NINA FEFFERMAN:**So, just to quickly 

### 5 follow up on that idea, I'm strongly in favor of the consumption 

### 6 per body weight idea. I also very much like, but I'm not sure 

### 7 how to integrate with that per body weight idea the idea of 

### 8 whole mass over your stage feeding, because your weight changes 

### 9 substantially over the course of being a larva at different 

### 10 days. So, there are good ways to do it, but one should be 

### 11 chosen and it should probably be discussed quite carefully how 

### 12 to do that. 

### 13 **DR. DANIEL SCHLENK:**Yes, Dr. Ostiguy? 

### 14 **DR. NANCY OSTIGUY:**This is Nancy Ostiguy. 

### 15 This question actually is looking at contact exposure. So, we 

### 16 may want to look at instead of a milligram per kilogram, a 

### 17 surface area measurement if we're looking at contact. 

### 18 **DR. DANIEL SCHLENK:**Okay. Jumping the gun a 

### 19 little bit I guess. The following questions, I'm thinking. 

### 20 Yes. In terms of diet, yes. Dr. Tarpy? 

### 21 **DR. DAVID TARPY:**To answer Dr. Fefferman's 

### 22 point, to reconcile those things, one is larval and the other is 

### 23 adult. So, I think it'd be pretty much impossible, I think 

### 24 logistically to measure the weight of larvae effectively, 

### 25 especially as it changes over time. So, it's not a constant. 

### 26 But for adults, they don't grow, so that

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1 standardizing by the adult exposure by weight would be possible, 

### 2 but then the total consumption over the feeding lifetime of the 

### 3 larvae would be the way I think would do it. Again, I think 

### 4 this will come up probably again in question number 8. 

### 5 **DR. DANIEL SCHLENK:**You guys have any other 

### 6 comments from the Panel before I turn it over to the agency? 

### 7 Okay. Any questions or clarification? Ms. Garber? 

### 8 **MS. KRIS GARBER:**Yes, Kris Garber. Dr. 

### 9 Schwab, you mentioned that you had some concerns about the 

### 10 assumption of linear scaling for application rate. I'm curious 

### 11 if you have an alternate suggestion to the linear assumption? 

### 12 **DR. PAUL SCHWAB:**No, not really. I can't say 

### 13 that I have something that is superior to that, but that is 

### 14 definitely an inherent assumption that it is all linear and it 

### 15 may or may not be. 

### 16 **DR. DANIEL SCHLENK:**Okay. So, we are all 

### 17 good with (a)? Oh, sorry, Mr. Pistorius has another comment 

### 18 here. 

### 19 **MR. JENS PISTORIUS:**Sorry, I was a little bit 

### 20 late. I just wanted to add, we calculated on this approach and 

### 21 it is actually in line with the HQ data and also the incident 

### 22 data from exposures, so I think this approach, as it was said 

### 23 before, it quite conservative. 

### 24 **DR. DANIEL SCHLENK:**Okay. This is for -- Dr. 

### 25 Schwab, did you get his comments? 

### 26 **DR. PAUL SCHWAB:**Not thoroughly, can you

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1 write that up for me? 

### 2 **DR. DANIEL SCHLENK:**All right. Let's go to 

### 3 (b) if we can, and then we will take a break. 

### 4 **MS. KRIS GARBER:**Kris Garber. Question 4, 

### 5 part (b) - please comment on the potential utility of the T-REX 

### 6 upper-bound residue value, i.e. 12 microgram a.i. per bee per 1 

### 7 pound a.i. per acre for a broader number of arthropod species to 

### 8 represent contact exposures to honey bees and to other insect 

### 9 pollinators that are directly sprayed with pesticides. 

### 10 **DR. DANIEL SCHLENK:**Okay. Dr. Schwab? 

### 11 **DR. PAUL SCHWAB:**This approach is sort of a 

### 12 modeling approach that's being used when there is an absence of 

### 13 hard data. Obviously, the direct studies of any sort of bees or 

### 14 pollinators would be preferred, but in the absence of data, this 

### 15 would be a method that could be used. One of the strengths of 

### 16 this that the data used in generating T-REX represents actual 

### 17 measurements on arthropods and are expected to actually reflect 

### 18 what you might see in honey bees and non-Apis bees. 

### 19 Some of the weaknesses would include that, 

### 20 although it was based on 14 studies, it was only done on a few 

### 21 different kinds - actually 2 different categories of pesticides, 

### 22 the carbamates and the organophosphates. Some of the others 

### 23 like the neonicotinoids - gee, I probably couldn't pronounce my 

### 24 own name at the moment - with totally different chemistry 

### 25 perhaps may not be very well reflected in this model. 

### 26 Let's see, doses and residues are based on

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1 normalization. Again, this gets back to the same thing. I 

### 2 don't have a better way to go, but it does assume that 

### 3 linearity. That was the sum of my comments at this point. 

### 4 **DR. DANIEL SCHLENK:**Dr. Potter? 

### 5 **DR. THOMAS POTTER:**I will say I agree with 

### 6 Dr. Schwab in regards to the points and the applicability of the 

### 7 study. Again to repeat, it would certainly be appropriate to 

### 8 have measurements on the target organism rather than have to 

### 9 extrapolate to others. With that said, I'm not sure that the 

### 10 arthropod data is any more uncertain that the traits for 

### 11 estimate. They are both, I think, highly uncertain. I think it 

### 12 is clearly a data gap that could be filled relatively easy 

### 13 without substantial cost. So, I certainly would want to 

### 14 emphasize that that be done. 

### 15 I will add perhaps there is another way of 

### 16 thinking about this in terms of estimating exposure, and that is 

### 17 simply to take the assumed spray rate, which is 1 pound per acre 

### 18 or 1 kilogram per hectare, if you do the arithmetic on that, 

### 19 that becomes 11 micrograms per square centimeter. Hopefully my 

### 20 arithmetic is working today, but I believe that is correct. So, 

### 21 that's the application rate for square centimeter. If we look 

### 22 in the literature, and I did some digging on this and I actually 

### 23 found this notation. I was pleased, which provided the surface 

### 24 area of a honey bee. 

### 25 Its these guys, Steve Roberts and John 

### 26 Harrison, maybe you know them whether they have been working in

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1 the field for a number of years, but this was an article 

### 2 published in the Journal of Experimental Biology in 1999. They 

### 3 pulled some wings off the bees. That sounded kind of gruesome, 

### 4 but anyway, the pulled the wings off and measured the surface 

### 5 area. They came up with 2.4 square centimeters. So, if you 

### 6 take your 11 micrograms per square centimeter and divide it by 

### 7 the 2.4, you get essentially a potential contact rate. That is, 

### 8 in essence, a 45 percent interception rate, if you will, of the 

### 9 chemical. So, that in some ways, I think it provides, you know, 

### 10 perhaps another back check in terms of what might be the 

### 11 reasonableness of this number, this 12 microgram per bee number. 

### 12 Perhaps there are some other folks who are much more familiar 

### 13 with bees in the room than I am can comment about the surface 

### 14 area as an issue, but I think that that at least provides a test 

### 15 of reasonableness to the value. 

### 16 **DR. DANIEL SCHLENK:**Yes, we will go with Dr. 

### 17 Fefferman first and then Dr. James. 

### 18 **DR. NINA FEFFERMAN:**Sorry, this may be way 

### 19 too detailed, but in terms of surface area of bees, there are 

### 20 some regions of bees that they (inaudible) much more frequently 

### 21 than others, so I don't know if you want to default the entire 

### 22 surface of your bee unless it's actually sort of an absorption 

### 23 through cuticle kind of thing. I don't know if bees do that. 

### 24 **DR. DANIEL SCHLENK:**Dr. James? 

### 25 **DR. ROSALIND JAMES:**This is Rosalind James. 

### 26 Maybe that would be like your upper limit of what they could

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1 possibly be exposed to. Unless you are spraying soil, a field 

### 2 is actually three-dimensional, so that pesticide rate is being 

### 3 spread over plant leaves and mostly on the upper and not so much 

### 4 on the lower. And in addition to that, bees are designed to 

### 5 collect things, and they are full of hairs, and they may 

### 6 actually collect more liquid than their surface area. So, 

### 7 contrary to what you would say, I would say the opposite that 

### 8 they have the surface tension and are made to gather things. 

### 9 So, that would be a very rough estimate I would say. 

### 10 **DR. THOMAS POTTER:**I will say that, but 

### 11 again, it's in line with the upper-bound estimate that is being 

### 12 proposed in the White Paper. So, that's why I put it forward as 

### 13 to say there is maybe one other way to think about this and that 

### 14 is indeed looking at surface area and potential for 

### 15 interception. I agree with all the things you said and 

### 16 certainly not being a honey bee physiologist, or bee 

### 17 physiologist at all, I have to default to that. But, you know, 

### 18 perhaps there is some more thought that can go into this 

### 19 generalized area, which might help us get some appreciation for 

### 20 interception rate, essentially is what we are talking about 

### 21 here. 

### 22 **DR. DANIEL SCHLENK:**Dr. Ostiguy? 

### 23 **DR. NANCY OSTIGUY:**This has come up a little 

### 24 bit before, but it shows up here very specifically. We are 

### 25 looking at exposures to honey bees and to other insect 

### 26 pollinators. As Dr. Berenbaum said yesterday, there are other

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1 pollinators than bees. We need to be clear what we are talking 

### 2 about. So are we talking about honey bees or are we talking 

### 3 about honey bees and other bee pollinators or are we including 

### 4 flies, or who are we including? 

### 5 **DR. DANIEL SCHLENK:**Okay. Any other comments 

### 6 from Panel members? It sounds like there's a proposed check 

### 7 that's out there with some caveats obviously. 

### 8 **DR. THOMAS POTTER:**I get carried away here. 

### 9 Tom Potter here. I'm sorry about that. Yes, you know, it's 

### 10 perhaps another check. It's a factor of two greater than the 

### 11 proposed value that's been suggested in the White Paper, which 

### 12 you know - and essentially we're talking about a 100 percent 

### 13 interception here. So, in that sense, I would think it puts the 

### 14 value that we are talking about in this case is probably 

### 15 reasonably conservative. 

### 16 **DR. DANIEL SCHLENK:**Any other comments from 

### 17 the Panel? Okay. Let me go to the Agency. Any questions or 

### 18 clarification? 

### 19 **MS. KRIS GARBER:**This is Kris Garber. I just 

### 20 want to just summarize what I heard back and make sure that I 

### 21 heard correctly. This is going to pertain to the whole 

### 22 question. So, it sounds like the proposed value of 2.7 

### 23 microgram a.i. per bee normalized to 1 pound a.i. per acre is a 

### 24 reasonable approach and that T-REX wouldn't necessarily provide 

### 25 an alternative or a better alternative, and that this surface 

### 26 area approach could be another way to just kind of like another

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1 line of evidence to support the use of the 2.7? But you're not 

### 2 necessarily proposing to use the 2.7 to calculate - I mean, 

### 3 you're not necessarily proposing to use the surface area 

### 4 approach to calculate risk quotients. 

### 5 **DR. THOMAS POTTER:**Not at this moment, no. 

### 6 This is Tom Potter here. Maybe Arthur had initial response 

### 7 here. I just wanted to leap in on the surface here. I simply 

### 8 put that as, you know, as another way of thinking about the 

### 9 problem and helping to define what the maximum exposure. I feel 

### 10 that whether or not the T-REX 2.7 value or the arthropod 12 

### 11 value, which one is better - I don't think we have the available 

### 12 information at this point to say. I would strongly emphasize 

### 13 that studies need to be done, collecting bees in sprayed fields 

### 14 and making residue measurements. I believe that that is the 

### 15 most appropriate path forward here. 

### 16 **DR. DANIEL SCHLENK:**Okay. Are we good? All 

### 17 right, let's go ahead and break for 15 minutes. Let's try to be 

### 18 back at 2:30. 

### 19 (WHEREUPON, a recess was taken) 

### 20 Go ahead and get going? We can? All right, 

### 21 Kris are you going to read 5 in? 

### 22 **MS. KRIS GARBER:**Yes. This is Kris Garber 

### 23 again. Charge question 5, part A - although bee larvae 

### 24 typically consume processed foods in the form of royal jelly and 

### 25 brood food throughout much of their development, they also 

### 26 consume honey and pollen during the last two days of the

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1 uncapped period. Please comment on the proposed use of nectar 

### 2 and pollen consumption rate of larvae during the last day of the 

### 3 larval developmental stage. Please include a discussion of the 

### 4 conservatism, strengths and limitations of this approach as well 

### 5 as a discussion of how this value may or may not correspond to 

### 6 data generated from larval toxicity endpoints. 

### 7 **DR. DANIEL SCHLENK:**Okay. Our lead 

### 8 discussant on that is Dr. Pettis. 

### 9 **DR. JEFF PETTIS:**Yes, Jeff Pettis. I'll lead 

### 10 with a question that we have for EPA about the data presented 

### 11 where royal jelly concentrations were graded 100 times lower 

### 12 than that in food consumed. I just like - in a few minutes when 

### 13 EPA has a chance to respond - to talk about those studies 

### 14 whether they've been published, whether they're a single study 

### 15 or multiple data points. I would like some clarification 

### 16 because they affect our response. 

### 17 But in general, we agree that unprocessed 

### 18 nectar and pollen would represent a worst case scenario. If we 

### 19 accept that, then they should be a conservative estimate by 

### 20 using them over brood food or royal jelly. 

### 21 As far as relevance to the larval toxicity 

### 22 endpoints, if you are doing larval assays, most of those are 

### 23 done with dilute royal jelly and so, the use of nectar and 

### 24 pollen again would be a conservative estimate and may not lead 

### 25 directly to larval endpoints. Most of those larval endpoints 

### 26 are done by spiking the royal jelly or the brood food. So they

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1 provide a valid assay tool for looking at larval toxicity 

### 2 issues. 

### 3 We see no issue with the use of dilute royal 

### 4 jelly in these larval assays. Some of the larval assay data 

### 5 that I've seen generated shows that when using brood food or 

### 6 dilute royal jelly, the adults produced in these assays lived as 

### 7 long and foraged normally as would workers reared in whole 

### 8 hives. So, those larval assays are valid, and again back to the 

### 9 use of nectar and pollen, I think they've produced a 

### 10 conservative estimate. So, that's just for 5A. 

### 11 **DR. DANIEL SCHLENK:**Dr. Ostiguy? 

### 12 **DR. NANCY OSTIGUY:**I concur. 

### 13 **DR. DANIEL SCHLENK:**Okay. Other panel 

### 14 members? Yes, Dr. Tarpy? 

### 15 **DR. DAVID TARPY:**This is Dave Tarpy. Jeff, I 

### 16 know that your lab has experience with these larval bioassays, 

### 17 but there is a lot of other labs that have slightly different 

### 18 approaches and in some cases, quite different approaches. So, 

### 19 how those translate to each other is, I think, something of 

### 20 consideration. So, is there an inherent assumption here of 

### 21 always having a control group in these bioassays to make sure 

### 22 that these things are relativized to different concentrations of 

### 23 the compound. 

### 24 **DR. JEFF PETTIS:**Jeff Pettis. I guess - I 

### 25 think there is a need for standardization within these larval 

### 26 assays. I know there has been some ring testing in the EU to

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1 try to standardize and make these larval assays valid. I don't 

### 2 think we are quite there yet with most of the labs in the U.S. 

### 3 **DR. DANIEL SCHLENK:**Dr. James? 

### 4 **DR. ROSALIND JAMES:**But at the same time, 

### 5 larval assays are exceedingly important. I mean, so we have to 

### 6 find a way to do them. 

### 7 **DR. DANIEL SCHLENK:**Mr. Pistorius? 

### 8 **MR. JENS PISTORIUS:**I would like to add to 

### 9 the comment of Jeff, that it is the same in the EU. I think 

### 10 most labs are not currently there yet. I have a question, or 

### 11 maybe a concern or I'm not sure what it is - maybe we will also 

### 12 discuss it later in the Aupinel section of question 9, but now 

### 13 when I read this here, I read that the concentration in royal 

### 14 jelly is in fact 100 lower. I think that somewhere it should be 

### 15 discussed if we do such a larva toxicity test, but maybe this is 

### 16 too early to ask. Are we actually using the lower 

### 17 concentrations, 100 times lower? If you do a risk assessment 

### 18 refinement, you have residue data in nectar and pollen, do you 

### 19 then say we go for concentration 100 times lower? Then again, I 

### 20 apologize if it's not appropriate to ask this at this time. 

### 21 **DR. DANIEL SCHLENK:**Well, I think that's up 

### 22 to Dr. Pettis if he thinks it appropriate to answer that 

### 23 particular question with that sort of component. Would you 

### 24 rather do that now or wait, I guess? 

### 25 **DR. JEFF PETTIS:**I think I would like to 

### 26 address it - Jeff Pettis. I'm hoping that the idea would be not

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1 to use 100 times lower value, but to use more of a worst case 

### 2 scenario in a larval assay. 

### 3 **MR. JENS PISTORIUS:**Agreed. 

### 4 **DR. DANIEL SCHLENK:**okay. Sounds good. Any 

### 5 other comments from the Panel for A? Okay. We go to the 

### 6 Agency. Are you clear with that response? 

### 7 **MS. KRIS GARBER:**Yes, I actually do have one 

### 8 clarifying question though. I can also address Dr. Pettis's 

### 9 question about the 100 times factor. So, Dr. James, you have 

### 10 mentioned earlier that we should consider accounting for dose 

### 11 over the entire larval life stage. So, would that entail 

### 12 expanding the consumption rate out to the fourth and fifth days 

### 13 or all five days? 

### 14 **DR. ROSALIND JAMES:**If I remember right, your 

### 15 data was actually on the entire food consumed, and then you had 

### 16 to extrapolate out to a daily rate - right? 

### 17 **MS. KRIS GARBER:**This is Kris Garber. So to 

### 18 clarify, it was broken out by the first three days and then food 

### 19 consumption over the fourth and fifth days. 

### 20 **DR. ROSALIND JAMES:**I would say the whole 

### 21 life stage. The royal jelly for the first three days and then 

### 22 the pollen and nectar consumed during the fourth and fifth or 

### 23 out to the seventh, depending on whether you are talking about 

### 24 workers or drones. I might as well say this here now - you sort 

### 25 of ruled out royal jelly because it had lower pesticide 

### 26 concentration in it, but if I remember right, that was based on

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1 one study. Is that right? 

### 2 **MS. KRIS GARBER:**So, this gets to Dr. 

### 3 Pettis's question also. It was based on two studies. The first 

### 4 one is actually a published study involving dimethoate and 

### 5 carbofuran. I believe the authors were Davis and Shuel, and 

### 6 it's cited on appendix 1, so you can confirm that. Actually, 

### 7 they observed concentrations that were more than 100 times 

### 8 different. I think it was a factor of 1000? And then the 

### 9 second study - those results are unpublished. Part of the 

### 10 results - the analysis was conducted by EPA, and that involved 

### 11 Imidacloprid. That was actually the basis for the 100X factor. 

### 12 So, nurse bees were fed concentrations of food that were 100 

### 13 times more than what showed up in the royal jelly that they were 

### 14 producing. 

### 15 **DR. DANIEL SCHLENK:**Just hold on a second. 

### 16 So, Dr. Pettis does that answer your question? 

### 17 **DR. JEFF PETTIS:**That does answer the 

### 18 question. 

### 19 **DR. DANIEL SCHLENK:**Okay. Great. And then 

### 20 Dr. Berenbaum, you have another comment? 

### 21 **DR. MAY BERENBAUM:**Just to say that this does 

### 22 come up in question 10, that the royal jelly estimates are based 

### 23 on one, but also there are some issues about the relative 

### 24 nutritional value and contribution, even if discounting the 

### 25 consumption of royal jelly ignores the relative impact on 

### 26 growth, but that may be more relative for question 10 right now.

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1 2 **DR. DANIEL SCHLENK:**Let's wait for question 

### 3 10, yeah, if we can. Any other 5A questions? Any other 5A 

### 4 comments? Okay. Let's move to the second letter of the 

### 5 alphabet. Somebody asleep out there. 

### 6 **MS. KRIS GARBER:**This is Kris Garber. Charge 

### 7 question 5, part B - please comment on the strengths and 

### 8 limitation of basing the Tier I screen for adult honey bees on 

### 9 food consumption rates of nectar foraging bees, including a 

### 10 discussion of the conservatism of this approach and how it 

### 11 relates to other types of worker bees and castes. 

### 12 **DR. DANIEL SCHLENK:**Okay. Dr. Pettis? 

### 13 **DR. JEFF PETTIS:**In general, I think the use 

### 14 of nectar consumption is generally appropriate, but may not be 

### 15 reflective of the age of the bees, which are often tested in 

### 16 Tier I. Tier I testing could use either randomly aged bees or 

### 17 young bees, and especially if they are young bees or newly 

### 18 emerged bees, then their diet normally would contain high 

### 19 amounts of pollen or pollen intake. So the use of just nectar 

### 20 consumption information may not be as appropriate as we would 

### 21 like. 

### 22 As far as relative to worker bees and other 

### 23 castes, as was brought up earlier by Dr. Tarpy, using weight 

### 24 might be a better estimate accounting for caste differences with 

### 25 the exception of the queen as I have already pointed out. But I 

### 26 do think that in general, worker bees - obviously they are

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1 easier to manipulate, they're available year round, so their use 

### 2 in Tier 1 screening is appropriate. With the exception of the 

### 3 pollen intake for young bees, it may not be reflective of the 

### 4 nature progression of the worker bee development. So, if you 

### 5 don't include some protein intake into the Tier I testing, which 

### 6 I don't think is normally the case, then I think we are missing 

### 7 an important component of their lifespan. That's all I have for 

### 8 that. 

### 9 **DR. DANIEL SCHLENK:**Okay. Dr. Ostiguy? 

### 10 **DR. NANCY OSTIGUY:**I concur. 

### 11 **DR. DANIEL SCHLENK:**Okay. Other Panel 

### 12 members? Dr. James? 

### 13 **DR. ROSALIND JAMES:**Just to reiterate about 

### 14 using honey bees as a surrogate for other bees, the non-Apis 

### 15 bees, all of the bees are going to probably eat pollen in 

### 16 addition to nectar. You can think of perhaps, every bee as a 

### 17 queen in the solitary bee. So, just one more argument for 

### 18 including pollen in the diet. It really maybe only matters if 

### 19 you have a difference in pesticide exposure between pollen and 

### 20 nectar. 

### 21 **DR. DANIEL SCHLENK:**Anyone else. Yes, Dr. 

### 22 Hunt? 

### 23 **DR. GREG HUNT:**I would just like to say I 

### 24 agree that including pollen is important because if you don't, 

### 25 it's an artificially high stress on the bee. 

### 26 **DR. DANIEL SCHLENK:**Okay. Dr. Pettis?

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1 **DR. JEFF PETTIS:**And just one other thought 

### 2 that occurred to me during the discussion was that with some of 

### 3 the systemics, the residue levels in pollen are generally higher 

### 4 than they are nectar. I think that has already been taken into 

### 5 account, but again, pollen consumption is normal for young bees. 

### 6 7 **DR. DANIEL SCHLENK:**Anyone else? Okay. Ms. 

### 8 Garber is that - any questions or clarification? 

### 9 **MS. KRIS GARBER:**I do have one point of 

### 10 clarification. I know that it's often difficult to separate out 

### 11 the exposure and effects components of the process, but I just 

### 12 want to make sure that I understand that the responses are 

### 13 specific to the exposure side so the consumption - just make 

### 14 sure that we're not talking about the toxicity tests or the 

### 15 conduct of the toxicity test. Is that correct? 

### 16 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 17 **DR. JEFF PETTIS:**Could you repeat the - 

### 18 because I'm not - go ahead. 

### 19 **MS. KRIS GARBER:**Okay. Thank you. Kris 

### 20 Garber again. So, the focus of this question is on the 

### 21 consumptions rates that we would use to estimate exposure in the 

### 22 Tier I analysis, and I know there is some overlap with how it 

### 23 may be related to the toxicity side of the risk quotient 

### 24 equation. I just want to make sure that the comments aren't 

### 25 focused on giving us advice on how to conduct the toxicity tests 

### 26 themselves.

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1 **DR. JEFF PETTIS:**Your point is well taken. I 

### 2 think there was some overlap in that, so yeah, you're right. If 

### 3 you're trying to look at just exposure, then we have different - 

### 4 some of the comments about pollen consumption and all were maybe 

### 5 not as appropriate as they might have been. 

### 6 **DR. DANIEL SCHLENK:**Okay, Dr. James? 

### 7 **DR. ROSALIND JAMES:**Along those lines, it 

### 8 mainly matters if there is a difference in past site 

### 9 concentration in the pollen versus the nectar. And there's a 

### 10 question that still seems to be hanging in the air. So, until 

### 11 there is more hard data on that, you do sort of need to keep the 

### 12 pollen and nectar ratio in mind when you are doing the 

### 13 calculations I think. 

### 14 **DR. DANIEL SCHLENK:**Dr. Ostiguy? 

### 15 **DR. NANCY OSTIGUY:**That actually brings up 

### 16 something that I have been thinking about and that is an area 

### 17 where we have a data gap. Half lives of our pesticides are 

### 18 calculated usually soil, water, et cetera, we don't necessarily 

### 19 know what the half life would be in the different matrices 

### 20 within the hive. That to me is a data gap. For us to be able 

### 21 to actually then extrapolate with our models and say yes, that's 

### 22 an appropriate exposure. 

### 23 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 24 **DR. MAY BERENBAUM:**And in terms of exposure 

### 25 too, one of the issues of using limited amount of data to 

### 26 estimate pesticide concentrations particularly when they are

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1 older studies is that today's chemistries are very different 

### 2 from old chemistries, so we really don't know whether they're 

### 3 comparable. So, that is why it's an issue of whether it would 

### 4 be another data gap in terms of estimating the degree of 

### 5 exposure through foods like royal jelly. We just don't have 

### 6 those data. 

### 7 **DR. DANIEL SCHLENK:**Okay. Any other comments 

### 8 for 5B? Are you good with 5B? Yeah? Okay. The Agency is 

### 9 nodding their heads. All right, let's read 5C into the record 

### 10 then. 

### 11 **MS. KRIS GARBER:**Kris Garber. Charge 

### 12 question 5C - please comment on the assumption that exposures 

### 13 through consumption of nectar and pollen are conservatively 

### 14 representations of potential exposures through consumption of 

### 15 honey, bee bread, brood food and royal jelly, all of which 

### 16 represent processed foods. 

### 17 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 18 **DR. JEFF PETTIS:**Our general assumption is 

### 19 that we agree with this that in fact, nectar and pollen would 

### 20 represent a worst case scenario or more so than processed food. 

### 21 As example, you bring out the royal jelly - it's a processed 

### 22 food - goes through the honey bee and there are processed 

### 23 intraglandular secretions, so even though bee bread and other 

### 24 stored products would undergo some degradation, so yes, we agree 

### 25 that it's the worst case scenario. 

### 26 **DR. DANIEL SCHLENK:**Okay. Dr. Ostiguy?

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1 **DR. NANCY OSTIGUY:**This is actually one 

### 2 situation where our extrapolation to native bees might not be as 

### 3 far because they're actually provisioned as larva with pollen 

### 4 and potentially a little bit of nectar. So, they're not getting 

### 5 brood food or royal jelly. 

### 6 **DR. DANIEL SCHLENK:**Okay. Dr. Berenbaum? 

### 7 **DR. MAY BERENBAUM:**Does anyone actually know 

### 8 the degree to which processing effects pesticide concentrations? 

### 9 I know of only one case in the literature, and it's degradation 

### 10 of phytochemicals in nectar or honey as it cures, but I don't 

### 11 know of any studies that have shown degradation of pesticides. 

### 12 **DR. DANIEL SCHLENK:**Sounds like a data gap. 

### 13 Okay. I guess we want to put that in the response as a data 

### 14 gap. Yeah, Dr. James? 

### 15 **DR. ROSALIND JAMES:**So, this is Rosalind 

### 16 James. May, would you say it's a reasonable assumption that it 

### 17 the concentration would not increase with processing? 

### 18 **DR. MAY BERENBAUM:**Well, it increases in 

### 19 nectar. It increases when you go from nectar to honey because 

### 20 it's concentration. It goes from, you know 80 percent water to 

### 21 18 percent water, so that is an increase in terms of what 

### 22 happens as it is stored in the hive. I don't think anybody 

### 23 knows. It is not going to increase, presumably, although there 

### 24 are some pesticides like organophosphates that can be 

### 25 bio-activated. Whether or not they are in honey or bee bread, I 

### 26 don't know if anybody would know. I'm not saying it's a good or

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1 bad - it's not an adequate proxy, I just don't know. 

### 2 **DR. DANIEL SCHLENK:**Dr. Sandy? 

### 3 **DR. MARTHA SANDY:**I think another data gap is 

### 4 we don't know what the degradation or metabolic products, 

### 5 whether they might be more toxic than the parent compound to the 

### 6 bees. 

### 7 **DR. DANIEL SCHLENK:**Yes, Mr. Pistorius? 

### 8 **MR. JENS PISTORIUS:**I don't think that 

### 9 although in the first moments on logic, when you go from nectar 

### 10 to honey that you would have a concentration of the residues, 

### 11 but I think data that we have and that were generated in EU has 

### 12 shown that it's actually vice versa, that you have a lower 

### 13 concentration in honey than nectar. And does data, for 

### 14 instance, also opinion which undermines it? 

### 15 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 16 **DR. MAY BERENBAUM:**Is that published? 

### 17 **MR. JENS PISTORIUS:**Yes, those are published 

### 18 data, for instance, in the Scientific Opinion 2012. There is a 

### 19 list of data that was used. 

### 20 **DR. DANIEL SCHLENK:**Mr. Pistorius. Dr. 

### 21 Fefferman? 

### 22 **DR. NINA FEFFERMAN:**Sorry, just for my own 

### 23 clarification, is that given the assumed half life of the active 

### 24 agents in the pesticide over that duration of time that it takes 

### 25 to reduce to the honey irrespective of honey reduction, or is it 

### 26 the action of reducing the nectar to the honey that you think is

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1 doing that? 

### 2 **MR. JENS PISTORIUS:**I'm not quite sure what 

### 3 is the reason for it? There has been data from (inaudible) who 

### 4 have shown that actually when the bees incorporate nectar and 

### 5 then they have it in the honey they still make themselves that 

### 6 actually after a few hours, the concentration is lower. So, the 

### 7 bee seems to absorb a little bit, but that's only one process. 

### 8 There seems to be another process that over time, and here we 

### 9 only have data - we have measurements of nectar directly when 

### 10 the foragers come back home, and then nectar in the hive and 

### 11 then this nectar to honey again. You can see that the 

### 12 concentrations get lower and lower, but I'm not quite sure what 

### 13 the exact process is behind it. 

### 14 **DR. DANIEL SCHLENK:**So, that would support 

### 15 conservation in terms of consumption of nectar and pollen, 

### 16 right? Yes. 

### 17 **DR. MAY BERENBAUM:**May Berenbaum. Can we 

### 18 include that in our - can we include that reference in our. 

### 19 **DR. DANIEL SCHLENK:**Definitely. I would 

### 20 think that would support exactly what Dr. Pettis initially said, 

### 21 yeah. Any other comments? Okay. Dr. Pettis? 

### 22 **DR. JEFF PETTIS:**I just would like to ask the 

### 23 group if they kind of agree with the things that are processed, 

### 24 the royal jelly or brood food, if there is general consensus 

### 25 that those values would be lower. It sounds like with honey, it 

### 26 could also be lower, but with pollen, there is a lot of level of

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1 uncertainty as far as stored pollen versus bee bread. 

### 2 **DR. DANIEL SCHLENK:**Okay. Everyone shake 

### 3 their heads. It looks good. Oh, question mark? No? Okay. 

### 4 Yeah, a lot of uncertainty there, it sounds like. Any other 

### 5 comments for 5C? Okay. Let me go Kristina - do you have any 

### 6 questions or clarification? 

### 7 **MS. KRIS GARBER:**No, we're good. 

### 8 **DR. DANIEL SCHLENK:**Okay. Great. All right, 

### 9 let's move on to charge question 6. Going to read letter A into 

### 10 the record? 

### 11 **MS. KRIS GARBER:**Kris Garber. Charge 

### 12 question 6, part A, foliar spray - please comment on the 

### 13 analysis presented in Section 3.1.1.1, with a focus on the 

### 14 extent to which the T-REX tall grass upper-bound residue may 

### 15 serve as an adequate surrogate to represent upper-bound 

### 16 pesticide concentrations in pollen and nectar of flowers that 

### 17 are directly sprayed with pesticides. 

### 18 **DR. DANIEL SCHLENK:**Okay. Our lead 

### 19 discussant on that is Dr. Schwab. 

### 20 **DR. PAUL SCHWAB:**Okay. Paul Schwab. The use 

### 21 of the T-REX model is necessitated due to the lack of data 

### 22 directly available on this subject. So, clearly data would be 

### 23 better, but when we look at the strengths of this, both in terms 

### 24 of the nectar and the pollen from the point of view of the 

### 25 nectar, T-REX is used currently by EPA. This is something that 

### 26 has been done for a while. It was based on actual experimental

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1 data as we spoke on the last question. The model actually uses 

### 2 very few input parameters, which helps in terms of its 

### 3 simplicity of use. It was developed with a large quantity of 

### 4 field data. The upper limit values are the upper-bound on 

### 5 residues form the actual measurements. 

### 6 The empirical data for nectar concentrations 

### 7 were compiled to evaluate the upper boundaries of T-REX and then 

### 8 specifically there were seven studies that were used. And, tall 

### 9 grass was chosen because it is three times higher than the 

### 10 upper-bound value in the empirical data. So, they were choosing 

### 11 from different species that were available and felt that the 

### 12 tall grass would be the best to sort of match the conservative 

### 13 approach that was being sought, and yet have as few as possible, 

### 14 if any, values of the empirical data exceed the tall grass 

### 15 value. 

### 16 From a pollen point of view, 11 studies for 

### 17 direct spray were used in developing for T-REX. The values that 

### 18 were compared to the empirical value show that the T-REX for 

### 19 tall grass is two times higher than the highest measure, so it 

### 20 is conservative. Twenty three actual data point were used in 

### 21 the development of this set, and the 110 milligrams per kilogram 

### 22 per pound of active ingredients per acre should be protected 

### 23 then. 

### 24 Some of the weaknesses would include that 

### 25 clearly no actual nectar or pollen data were used, so this is a 

### 26 very strong weakness, if that makes sense. Normalize to 1 pound

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1 per acre assumes a linear change, again sorry. The model does 

### 2 not contain actual values for pollen nectar from flowers that 

### 3 were directly sprayed. The highest empirical values correspond 

### 4 to a mean measurement of pesticides in nectar not a maximum. 

### 5 Therefore, the comparisons were made to T-REX as not an 

### 6 upper-bound really - this is just a matter of the way it was 

### 7 developed, really doesn't have to do with whether or not it's 

### 8 conservative. And, crop specific factors really would be 

### 9 difficult to account for under these circumstances. 

### 10 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 11 **DR. JEFF PETTIS:**I just have one additional 

### 12 comment or weakness. The T-REX model may in fact be 

### 13 conservative, but only if active ingredients are tested. They 

### 14 may not be predictive of the actual residues of the formulated 

### 15 product. So, if the formulated product was used, it would 

### 16 change the movement, penetration and longevity of the active 

### 17 ingredient in the field. So, that's a weakness of just using 

### 18 the a.i., the active ingredient. 

### 19 **DR. DANIEL SCHLENK:**Dr. Potter? 

### 20 **DR. THOMAS POTTER:**You know, I generally 

### 21 concur with both Dr. Schwab and Dr. Pettis. I think tall grass 

### 22 appears to be conservative, but I am not convinced that we have 

### 23 the data to determine whether the value is conservative or not. 

### 24 25 There is an appearance of conservatism, but 

### 26 the reality is -- and I've made this comment several times

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1 during the meeting in the form of questions - we have apparently 

### 2 a paucity of data, actual residue measurements on nectar and 

### 3 pollen with which to compare to this estimate. 

### 4 I wanted to make one comment about one of the 

### 5 papers, peer review papers that was used in the data 

### 6 compilation. That was the paper by Choudhary and Sharma. I 

### 7 took a quick read through it, and I think this is one of ones 

### 8 where maybe a little more peer review would have helped. 

### 9 I had some serious questions after I read 

### 10 through that paper. I still am puzzled as to how they came up 

### 11 with the detection limits that they reported. With regard to 

### 12 study design, the spray was applied when 50 percent of the 

### 13 flowers were open. There is no accounting for flowers opening 

### 14 after the spray was applied. The pollen and nectar were 

### 15 collected from bees that in theory were foraging in their crops, 

### 16 that they had treated, but there is no way of knowing that. 

### 17 So, we have a data quality problem in terms of 

### 18 study design. With that said, I think our subset of data that 

### 19 we have that it is reliable with which to compare to this T-REX 

### 20 value was even smaller unfortunately. So, I just wanted to make 

### 21 the comment about that and again, beat on this idea that we need 

### 22 a data call-in, need to get that information compiled, and 

### 23 that's going to, I think, give his whole risk assessment process 

### 24 in terms of dietary exposure a whole lot more credibility. 

### 25 One final point about nectar, because it's a 

### 26 liquid, was that it's roughly a 30 percent sugar solution.

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1 Pesticides do dissolve in it. Therefore, the solubility of 

### 2 pesticide active ingredients in the nectar ought to be 

### 3 considered in terms of assessing potential exposures. I think 

### 4 if we go through that process, we might gain some further 

### 5 insight into how conservative out estimates are. Again, I 

### 6 strongly emphasize we need to call in data, we need more 

### 7 information in order to be able to determine whether T-REX is an 

### 8 appropriate exposure estimate or not. Secondly, we perhaps 

### 9 ought to - in the case of nectar, which appears to be the 

### 10 primary exposure pathway, we need to take into account the 

### 11 physical and chemical properties of the active ingredients in 

### 12 terms of ultimately deriving our exposure estimates. 

### 13 **DR. DANIEL SCHLENK:**Any other panel members 

### 14 want to weigh in on this one? Dr. Hunt? 

### 15 **DR. GREG HUNT:**Yeah, I don't know if this 

### 16 data is relevant to Tom's question, but the managed pollinator, 

### 17 coordinate agricultural project has sentinel apiaries and in six 

### 18 different states, I believe, throughout the country and 

### 19 collecting all kinds of pesticide residue data on bee-collected 

### 20 pollen and wax and bees, and there are some other published 

### 21 studies that are looking at extant samples, so that may help to 

### 22 identify maximum levels that are seen. 

### 23 **DR. DANIEL SCHLENK:**Would you provide that 

### 24 information to Dr. Schwab, I guess, for his comments so that he 

### 25 can get that in there? Yes, Mr. Pistorius? 

### 26 **MR. JENS PISTORIUS:**Again, from the EFSA

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1 opinion or the EFSA opinion site that there are a few more 

### 2 studies that undermine some of the residue data that are 

### 3 mentioned here in Table 3 of the White Paper, so I completely 

### 4 agree and offer to send those data to compile them. 

### 5 **DR. DANIEL SCHLENK:**Thanks. Anyone else to 

### 6 comment? Okay. Go to Ms. Garber, any questions or 

### 7 clarification? No? Okay. Read in the letter that follows A. 

### 8 **MS. KRIS GARBER:**Kris Garber. Charge 

### 9 question 6, part B, soil application - please comment on the 

### 10 analysis presented in Section 3.1.2, with a focus on the extent 

### 11 to which the Briggs' model may generate estimates of pesticide 

### 12 exposure in plant stems that can represent upper-bound pesticide 

### 13 concentrations in pollen and nectar of flowers. 

### 14 **DR. DANIEL SCHLENK:**Dr. Schwab. 

### 15 **DR. PAUL SCHWAB:**Okay. Looking at this from 

### 16 the point of view of the strengths. This has been used in a 

### 17 couple of different approaches. It's used in the PRZM model. 

### 18 It has also been selected by United Kingdom Environmental Agency 

### 19 as coming closest to estimating empirical data. All the 

### 20 pesticides that were used in this model were nonionic. That's 

### 21 not necessarily a strength, but it does relate to an awful lot 

### 22 of the pesticides in question. 

### 23 The range in the Log Kow values that were used 

### 24 in this pretty well encompasses the range of Kows of various 

### 25 pesticides that we discussed thus far. The original model was 

### 26 based only on the Log Kow and regression analysis of

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1 experimental data, which made it fairly simple. Ryan et al. 

### 2 modified this and added three soil parameters to make the model 

### 3 more soil specific. They added the soil bulk density, so a 

### 4 water content, fraction of organic content in the soil. This it 

### 5 makes it then possible because the way that the model was 

### 6 developed was out of solution. This makes it possible then to 

### 7 do it out of the soil. It does require having these parameters, 

### 8 however. 

### 9 It was tested against experimental data that 

### 10 is by Briggs' on five chemicals. Modeling was conservative to 

### 11 maximize the concentrations in solution. Now the predicted data 

### 12 are within an order of magnitude to be experimental, but in the 

### 13 White Paper, a development was used to get the 95th percentile 

### 14 upper-bound estimates for the transportation stream 

### 15 concentration factors. This was discussed yesterday. 

### 16 Some of the weaknesses are that it is indeed a 

### 17 model. It does only examine nonionic compounds, so actually the 

### 18 mechanism of uptake could be completely different for ionizing 

### 19 species. The insecticides that were examined were carbamates 

### 20 and phenylureas, and based on only one kind of plant. This is 

### 21 definitely a drawback. 

### 22 The soil parameters that were used in the Ryan 

### 23 modification may or may not be known. Most particularly, the 

### 24 soil moisture content will be a huge variable. Bulk density 

### 25 needs to be known, but along with the fraction of the organic 

### 26 carbamate is very much spacially dependent both in terms of the

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1 horizontal and vertical directions. So, this can actually add 

### 2 some uncertainty to the whole process. I mean, it's a necessary 

### 3 step, but it doesn't really tighten up the model very much. 

### 4 Now the model is used for stem concentrations 

### 5 in the transportation stream and not for pollen or for nectar. 

### 6 So, it sort of gets back to the same problem we had with T-REX, 

### 7 in that we are examining one thing and extrapolating to another 

### 8 in terms of plant tissues. 

### 9 In testing the model against five chemicals 

### 10 with experimental data, various assumptions had to be made 

### 11 because this was not really using - well apparently these were 

### 12 unknown. So, there are multiple other assumptions. So, this 

### 13 would get into some uncertainty again. 

### 14 Not all the values predicted by Briggs' model 

### 15 were greater than the experimental values. This is why they 

### 16 went to the 95th percentile method, but even when this was done 

### 17 to try to gather the upper bound, still not all the data were 

### 18 above the empirical data, so it's not really truly conservative. 

### 19 20 I would like to add, some discussion has been 

### 21 forward both within the White Paper itself as well as other 

### 22 comments that this particular relationship is somewhat dated, 

### 23 somewhat limited. But, I would like to throw in that it was 

### 24 modified, reexamined and other data were put together by 

### 25 Burghardt and Schonherr in 1998 added another 10 or 15 years or 

### 26 so to updating this. These were a much broader array of

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1 chemicals used. Also a different plant species was used. So, 

### 2 this could really add to the dataset and help strengthen the 

### 3 model. That reference will be included in my comments. 

### 4 And finally, we need to put an upward bound on 

### 5 the water concentration of the pesticide determined by 

### 6 solubility of the compound in question. It kind of gets back to 

### 7 sort of the nectar question that Tom was talking about just a 

### 8 moment ago, in that we can't assume that we can get to just any 

### 9 water soluble or any water concentration, but that we must 

### 10 identify and upper bound and make sure that it's not exceeded 

### 11 because of the physical properties of these compounds. 

### 12 **DR. DANIEL SCHLENK:**Thank you. Dr. Pettis? 

### 13 **DR. JEFF PETTIS:**I agree with the comments of 

### 14 Dr. Schwab. I just want to elaborate a bit on one more aspect, 

### 15 and that is the use of stem data to extrapolate to nectar and 

### 16 pollen. I would agree in principle that it's a conservative 

### 17 estimate, and there are a few published studies out there, which 

### 18 examine stem, pollen and nectar, and almost all of those show 

### 19 that there are higher concentrations in the stem or leaf. So, I 

### 20 would agree that in general, that's a valid assumption. One 

### 21 caveat is that plant by plant could be quite different. So, 

### 22 trying to extrapolate to all plants from hardwood or few crop 

### 23 plants may not be valid. So it could be a crop-by-crop 

### 24 difference there. 

### 25 **DR. DANIEL SCHLENK:**Okay. Thanks. Dr. 

### 26 Potter?

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1 **DR. THOMAS POTTER:**I concur. I think that 

### 2 both Dr. Schwab and Dr. Pettis have, I think, hit on the points. 

### 3 I generally agree that Briggs is a good first cut. I want to 

### 4 emphasize that there are substantial uncertainties when that 

### 5 model is applied to charge species and that needs to be taken 

### 6 into account. Indeed, there are many active ingredients, 

### 7 particularly in the herbicide realm these days, that are 

### 8 charged, in fact I think increasingly, we will see more of that. 

### 9 So, that's a major data gap that needs to be filled, not only 

### 10 for this risk assessment, but for the use of this same equation 

### 11 and many other pesticide fate and transport models. It has been 

### 12 propagated through the system, shall we say. So, there is 

### 13 substantial effort needed to update that model, particular for 

### 14 charge species. 

### 15 **DR. DANIEL SCHLENK:**Any other comments? 

### 16 Okay. Ms. Garber, any questions or clarification? 

### 17 **MS. KRIS GARBER:**No, we're good thank you. 

### 18 **DR. DANIEL SCHLENK:**Great. 

### 19 **DR. PAUL SCHWAB:**Dr. Schlenk, before we move 

### 20 on, may I request that we reverse the order of discussion on the 

### 21 next two? 

### 22 **DR. DANIEL SCHLENK:**Sure. 

### 23 **DR. PAUL SCHWAB:**I think that it would be a 

### 24 more natural discussion to talk about the 1 milligram per 

### 25 kilogram value before we compare that the Briggs' model. Would 

### 26 that be okay?

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1 **DR. DANIEL SCHLENK:**So you'd like to go with 

### 2 D first before C? 

### 3 **DR. PAUL SCHWAB:**Yes. 

### 4 **DR. DANIEL SCHLENK:**Okay. Sure. Go ahead 

### 5 and read D in, great. 

### 6 **MS. KRIS GARBER:**Kris Garber. Charge 

### 7 question 6, part D, seed treatments - please comment on the 

### 8 analysis presented in Section 3.1.3, including a discussion of 

### 9 the strengths and limitations of the use of 1 milligram a.i. per 

### 10 kilogram value as an upper-bound concentration for pollen and 

### 11 nectar of seed-treated crops. 

### 12 **DR. DANIEL SCHLENK:**Thanks. Dr. Schwab? 

### 13 **DR. PAUL SCHWAB:**Okay. This particular 

### 14 approach is addressing section 3.2.2 of the white paper, and 

### 15 quoting from that, it says that if pesticide specific residues 

### 16 in pollen and nectar are not available, a screening value of 1 

### 17 milligram a.i. per kilogram plant matrix is the assumed 

### 18 exposure. This is from EPPO 2010. This is based on the upper 

### 19 limit value from empirical data of pesticide concentrations and 

### 20 different plant parts. The strengths of this would be that it 

### 21 is stated that the 1 mg per kg value tends to be higher than the 

### 22 reported concentrations. Most pesticides tend to be about two 

### 23 orders of magnitude below this particular value. 

### 24 Obviously, a weakness of this is that because 

### 25 there are no data that this is just simply - it's not exactly 

### 26 random, but it is selecting a value rather than depending on

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1 data. We sort of visited this concept before and this is a real 

### 2 obvious problem here. 

### 3 The value may not be completely protective. 

### 4 The value really takes into account no real aspects of the real 

### 5 situations that are going on in the field, but that's why 

### 6 there's no data. 

### 7 **DR. DANIEL SCHLENK:**Okay. Dr. Pettis? 

### 8 **DR. JEFF PETTIS:**I concur. 

### 9 **DR. DANIEL SCHLENK:**And Dr. Potter? 

### 10 **DR. THOMAS POTTER:**I concur. 

### 11 **DR. DANIEL SCHLENK:**Wow. Any other comments? 

### 12 Okay. Ms. Garber? Are you good with that one? 

### 13 **MS. KRIS GARBER:**Good. Thank you. 

### 14 **DR. DANIEL SCHLENK:**So, then, you want to 

### 15 read in C next, is that correct? Okay. 

### 16 **MS. KRIS GARBER:**Charge question 6C, soil 

### 17 application - please discuss the relative strengths and 

### 18 limitations of the 1 milligram a.i. per kilogram value and the 

### 19 soil uptake model (the Briggs' model) proposed in the white 

### 20 paper as Tier I screens, including consideration of the extent 

### 21 to which this method may generate conservative Tier I estimates 

### 22 of dietary exposures to bees. Does the Panel conclude that the 

### 23 one approach may be better suited to specific types of 

### 24 assessment scenarios? If so, please elaborate. Alternatively, 

### 25 if both approaches are equally suited for a Tier I screen, 

### 26 please provide guidance on how to best capture variability and

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1 uncertainty in the exposure estimates using the two approaches. 

### 2 **DR. DANIEL SCHLENK:**And Dr. Schwab? 

### 3 **DR. PAUL SCHWAB:**Okay. There were three 

### 4 parts to this. One is to compare the two and the other is to 

### 5 try to decide if one is better suited to specific scenarios than 

### 6 the other, and finally then to provide guidance on capturing 

### 7 variability and uncertainty. 

### 8 We just reviewed the Briggs' model as well as 

### 9 the 1 milligram per kilogram level. That won't be repeated 

### 10 here, but EPPO approach is that if all positions - assuming that 

### 11 nothing is known about the soil, the plants or the pesticides - 

### 12 and this approach assumes that any attempt to estimate pesticide 

### 13 residues offers no more reliability or even unreliability than 

### 14 simply assigning a value. The Briggs' model makes an attempt to 

### 15 try to integrate information about the soils and the chemistry 

### 16 of the chemical in the plants, to try to make a projection of 

### 17 the concentrations. 

### 18 So, in this way, the Briggs' model might be a 

### 19 more suitable way to go. Unfortunately, it also has its 

### 20 uncertainties because of limitations of the number of chemicals 

### 21 that were involved and development of the model as well as the 

### 22 limited number of plant species. So this is clearly a problem. 

### 23 24 In terms of one approach being better than the 

### 25 other for a given scenario, I would assume then that the Briggs' 

### 26 model would be best used for those plant species that are

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1 closest to the - as modified also by Burghardt and Schonherr - 

### 2 best suited to those plant species that were used in the 

### 3 development of the model. We would have to investigate more, 

### 4 more information to see where this model broke down. So, just 

### 5 assigning a single value would be best. 

### 6 Then finally in terms of providing guidance on 

### 7 how to capture variability and uncertainty, there is so much 

### 8 uncertainty and it's really hard to pin this down. 

### 9 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 10 **DR. JEFF PETTIS:**I concur with the comments 

### 11 of Dr. Schwab. 

### 12 **DR. DANIEL SCHLENK:**Dr. Potter? 

### 13 **DR. THOMAS POTTER:**I concur. 

### 14 **DR. DANIEL SCHLENK:**Okay. Anyone else on the 

### 15 Panel? Ms. Garber, you clear with that one? 

### 16 **MS. KRIS GARBER:**Clear, thank you. 

### 17 **DR. DANIEL SCHLENK:**Great. Okay. So, last - 

### 18 let's get back to letter E, read that into the record please. 

### 19 **MS. KRIS GARBER:**Kris Garber, charge question 

### 20 6E - please comment on other approaches or data that should be 

### 21 considered for estimating upper-bound estimates of pesticide 

### 22 residues in pollen and nectar as a Tier I screening-level 

### 23 assessment for pesticides applied via foliar spray, soil 

### 24 application or seed treatment. 

### 25 **DR. DANIEL SCHLENK:**And Dr. Schwab? 

### 26 **DR. PAUL SCHWAB:**Well, just sort of quickly

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1 reviewing the methods that have been described in terms of 

### 2 trying to get a handle on these values, we have direct 

### 3 measurements of pesticides and questions on the very specific 

### 4 plant tissues of interest, whether they are nectar or pollen or 

### 5 whatever. Secondly, there would be direct measurement of the 

### 6 pesticide in question and other plant issues such as stems and 

### 7 leaves and so on. 

### 8 Third, direct measurement of surrogate 

### 9 pesticides in various plant tissues, then we have modeling 

### 10 approaches after foliar spray, such as the T-REX modeling, 

### 11 pesticide uptake from the soil, the Briggs'/Ryan/Burghardt 

### 12 approach to this. And then finally the sixth one that we looked 

### 13 at, assigning a default value with no other method is available. 

### 14 15 This is sort of an exhaustive list. There 

### 16 might be some other methods. There might be some other modeling 

### 17 methods, some purely mathematical kinds of approaches that might 

### 18 be taken, but the way I - at least in my own limited perspective 

### 19 on this would see this as just sort of tweaking what we already 

### 20 have. Other approaches might not be all that truly different. 

### 21 Some may disagree. 

### 22 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 23 **DR. JEFF PETTIS:**I agree. I guess I'd just 

### 24 like to emphasize that, yeah, we need direct measurements and 

### 25 real data, and maybe some of this data may be held by the 

### 26 registrants and could be useful in the evaluation process.

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1 **DR. DANIEL SCHLENK:**And Dr. Potter? 

### 2 **DR. THOMAS POTTER:**I will throw this out 

### 3 again that I think that perhaps an upper-bound on nectar has to 

### 4 do with the solubility of the chemical in the nectar solutions. 

### 5 I'm not aware of any systematic study looking at solubility of 

### 6 organic chemicals in nectar, but that's something that can be 

### 7 relatively easily rectified, and perhaps would be a useful data 

### 8 gap to fill. 

### 9 **DR. DANIEL SCHLENK:**Okay. Any other Panel 

### 10 members? Okay. Ms. Garber, are you good with question 6? 

### 11 **MS. KRIS GARBER:**Yes we are, thank you. 

### 12 **DR. DANIEL SCHLENK:**Okay. So, we are 

### 13 actually doing really well in terms of our schedule. Would I 

### 14 would like to do is go ahead and finish up the exposure side 

### 15 today. Maybe we'll take break after that and start the effects 

### 16 side tomorrow. So, the last exposure question really is 7. I 

### 17 know you guys haven't met together, but apparently, everybody 

### 18 has an answer, so we'll go the traditional route. You guys can 

### 19 go through that. You got it? Okay. So, if we can go ahead and 

### 20 read 7A into the record. It's way down there. Yeah, I think 

### 21 it's the next page perhaps - there it is. 

### 22 **MS. CHRISTINA WENDEL:**Christina Wendel, 7A - 

### 23 please comment on the strengths and limitations of basing the 

### 24 Tier I exposure method on contact and diet. Does the Panel 

### 25 agree that for the majority of pesticide applications, the 

### 26 primary exposure routes for bees will be represented by contact

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1 and diet? 

### 2 **DR. DANIEL SCHLENK:**And Dr. Potter, you are 

### 3 lead on this one. 

### 4 **DR. THOMAS POTTER:**Okay. And the way this 

### 5 question seems to broken out, I think we are going to have to 

### 6 have a short answer to the beginning and then we'll address the 

### 7 individual exposure pathways down in the subsequent questions. 

### 8 **DR. DANIEL SCHLENK:**Okay. 

### 9 **DR. THOMAS POTTER:**First and foremost, I 

### 10 would simply say that diet and direct contact are likely 

### 11 dominant exposure pathways. However, other exposure pathways 

### 12 have the potential to be quantitatively significant and should 

### 13 therefore be considered. 

### 14 **DR. DANIEL SCHLENK:**Okay. 

### 15 **DR. THOMAS POTTER:**And we've covered some of 

### 16 that earlier when we talked about the conceptual model, and we 

### 17 will get to some of the other issues on that further. 

### 18 **DR. DANIEL SCHLENK:**Okay. Dr. Hunt, I have 

### 19 you as your firs associate. 

### 20 **DR. GREG HUNT:**I don't have anything to add. 

### 21 22 **DR. DANIEL SCHLENK:**And Dr. Pettis? 

### 23 **DR JEFF PETTIS:**Just briefly, I agree that 

### 24 contact and diet may be the primary routes, and that may vary 

### 25 contact over diet or diet over contact by chemical type. Then, 

### 26 I don't think we should discount these other routes of exposure

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1 yet. We will get to those. 

### 2 **DR. DANIEL SCHLENK:**Other Panel members? 

### 3 Okay. I will go to the Agency. Are we clear with the answers 

### 4 for 7A? Okay. So let's go to the letter that precedes C. 

### 5 **MS. CHRISTINA WENDEL:**Question 7, subpart 

### 6 (b), dust - if the panel believes that this exposure route 

### 7 should be quantitatively included in the Tier I exposure method, 

### 8 excuse me, Christina Wendel -- for the relevant application 

### 9 types i.e., seed treatment, please discuss the data that may be 

### 10 needed to address the exposure route quantitatively. 

### 11 **DR. DANIEL SCHLENK:**And Dr. Potter? 

### 12 **DR. THOMAS POTTER:**Okay. Well first, I would 

### 13 like to propose a slight modification to the question here, 

### 14 which is we don't restrict it to seed treatments. Is that 

### 15 acceptable? 

### 16 **DR. DANIEL SCHLENK:**I think as long as you 

### 17 state that's how you answer the question, then you can put that 

### 18 in there. 

### 19 **DR. THOMAS POTTER:**Okay. And I say that 

### 20 because I think that there are basically two sources of 

### 21 pesticide laden dust that may be present in the environment of 

### 22 the bees. Certainly, emissions from seed treatments have been 

### 23 widely discussed. I believe Jens Pistorius is going to say 

### 24 something about that, so I won't get into detail on that. I 

### 25 think the data strongly indicates that it should be considered. 

### 26

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1 Ideally, we should have information that 

### 2 quantifies those admissions. You know, we've heard they exist, 

### 3 but not being a person who has conducted research in this area, 

### 4 I'm hard pressed to know what the emission rates are from 

### 5 planters. 

### 6 We've also heard that there are a number of 

### 7 mitigation measures that can be applied in modified planters and 

### 8 reducing emissions. Certainly, it's important and relevant that 

### 9 we get quantitative information about how effective those 

### 10 practices may or may not be. So, with regard to the seed 

### 11 coating, I think we again, the theme is we need more data and we 

### 12 are greedy for data here. 

### 13 The second part of the dust issue, which I'd 

### 14 like to raise, which hasn't come up in the discussion and didn't 

### 15 surface in the White Paper is the potential for dust to be 

### 16 generated by wind from treated fields. This is a remarkably 

### 17 little researched area, although I think it's potentially 

### 18 important. The limited data that are available in the published 

### 19 literature, I'm not aware of any unpublished registrant studies, 

### 20 indicate that in certain fine fractions of dust generated from 

### 21 fields, in particular something we define as PM10. Probably 

### 22 most of us in the room are familiar with that. 

### 23 You can have very strong enrichment of 

### 24 pesticide active ingredients. So dust, i.e. generated from 

### 25 wind-blown materials from fields, can have relatively high 

### 26 pesticide concentrations. Therefore, I think we perhaps need to

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1 step back and contemplate that a little bit and see if this may 

### 2 be an exposure pathway for bees, which would include a direct 

### 3 deposition of the dust on hives that are actually remote from 

### 4 the field as well as body contact from dust that accumulated on 

### 5 various surfaces, including plant surfaces. 

### 6 I strongly feel that there are two parts of 

### 7 the dust question that need to be examined and relative to 

### 8 potential exposures in this case. 

### 9 **DR. DANIEL SCHLENK:**Dr. Hunt: 

### 10 **DR. GREG HUNT:**Well, I'll just speak to the 

### 11 seed treatment. Neonicotinoid seed treatments in particular, in 

### 12 the conceptual model, this is modeled - the EPA White Paper is 

### 13 only looking primarily at systemic movement plant parts. But 

### 14 clearly, we're seeing a problem with dust, particularly with 

### 15 corn planting and in regards to the soil. We see at least twice 

### 16 the concentrations of Clothianidin that we find in corn pollen. 

### 17 We're seeing a lot of reports, many of which apparently aren't 

### 18 getting transmitted to the EPA, and I think there is a lag in 

### 19 that also because, for example, in Indiana, the office of the 

### 20 state chemist has looked at 14 incident reports and they all 

### 21 came up positive for Clothianidin. 

### 22 In Ohio, there was something like 50 reports, 

### 23 incident reports, which again have not gotten their way to the 

### 24 EPA. In Ontario and Quebec, there are a lot of positive reports 

### 25 - over 130 of them, I understand - just from this year. Last 

### 26 year in the midwest, there were no incident reports. The

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1 difference is that now the beekeepers know about it, so they're 

### 2 making reports. 

### 3 I think dust through talc and abrasion of seed 

### 4 is a mode of exposure that needs to be addressed. We need more 

### 5 data on residues and wax comb, which we have virtually nothing 

### 6 from yet, but we're working on that, and more pollen samples. 

### 7 So, I think I'll leave it at that. 

### 8 **DR. DANIEL SCHLENK:**Okay. And Dr. Pettis? 

### 9 **DR. JEFF PETTIS:**Just a few comments to add 

### 10 relative to dust, I agree that it's generated a lot of interest 

### 11 and public opinion of things this year. I don't think in 

### 12 general - there probably are two ways that dust could become an 

### 13 issue. One is the seed treatment and the other is just general 

### 14 dust from sprayed crops. 

### 15 I do feel though, however, as far as looking 

### 16 at exposure levels that even with corn and seed treatment that 

### 17 the dust exposure is going to be a temporary effect, temporary 

### 18 and short-term effect whereas the translocation throughout the 

### 19 plant and expression in say, corn pollen, would be more of a 

### 20 chronic effect. So, I don't want to sight of that in focusing 

### 21 on the dust. I do think dust is a real issue that should maybe 

### 22 have a solid box rather than a dotted box. In terms of relative 

### 23 to dietary exposure, I think we just need to keep in mind that 

### 24 the expression of systemics in pollen and nectar are probably of 

### 25 more concern. 

### 26 **DR. DANIEL SCHLENK:**Okay. Mr. Pistorius?

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1 **MR. JENS PISTORIUS:**Thank you. I think I 

### 2 have to have a longer comment on this. Looking at the question, 

### 3 I think there are two questions in which data should be a useful 

### 4 Tier I exposure method. The second part of the question is 

### 5 basically what data may be needed to address the exposure route 

### 6 quantitatively. 

### 7 From the research that we've done at the 

### 8 Governmental Institution in the last year, I think we can say, 

### 9 I've had the honor to make a presentation to EPA on the 27th of 

### 10 July last year. We presented a tip of the iceberg of all these 

### 11 data. We can say that dust is a relevant route, which has to be 

### 12 integrated into the risk assessment. Nevertheless, I think on 

### 13 Tier I, so that's the answer for the first part of the question, 

### 14 we don't need any more data. Although there are scientific 

### 15 question still open, for instance, that we have the indication 

### 16 that compared in gram a.i. per hectare for instance or the oral 

### 17 and contact toxicity compared with dust to spray. There is a 

### 18 higher problem, a higher toxicity of dust compared to sprays, 

### 19 also when we compare the effects of the sprays and dust, between 

### 20 a spray application and a dust application, looking at the same 

### 21 gram a.i. per hectare with higher effects for dust. 

### 22 Nevertheless, I think we have the option to 

### 23 say for Tier I screening exposure method, we can say that it is 

### 24 enough. If we look at the acute oral and contact data that we 

### 25 get with feeding bee nectar. So, we get an indication that, you 

### 26 know, we have highly toxic substances for instance. Then the

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1 question is what is out exposure? We have done a lot - and I 

### 2 think I must apologize, we haven't published all the data yet, 

### 3 we are still working on the science. We've done a lot on 

### 4 generating data of seed treatment quality, what does it mean, 

### 5 how can you improve it, then machinery, what does it mean, how 

### 6 does the culmination of the two, considering that you can use 

### 7 deflectors with different efficacy, translate into potential 

### 8 exposure off crop. 

### 9 With this off-crop exposure, we think is it 

### 10 really important issue that has to be considered. There would 

### 11 be data available that actually allow integration into the risk 

### 12 management process. What I believe is that the management 

### 13 issues that have been considered by EPA are an effective way of 

### 14 mitigating the risks if you have a quality assurance of the seed 

### 15 treatment quality and also the planting practices. 

### 16 But in general for this answer, I can only 

### 17 give a limited answer for the second part of the question. The 

### 18 data that may be needed to address the exposure route 

### 19 quantitatively, I think that for highly toxic substances, you 

### 20 may have to go for higher tier tests. 

### 21 **DR. DANIEL SCHLENK:**So, it sounds like a 

### 22 little bit of disagreement there. Some would say yes dust and 

### 23 Mr. Pistorius says no dust, I guess, in terms of the Tier I, is 

### 24 that what I hear? 

### 25 **MR. JENS PISTORIUS:**Jens Pistorius. I think 

### 26 what you can do in the oral and contact laboratory tests, you

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1 get an indication of how toxic is such a substance. If you have 

### 2 highly toxic substance, those are a matter of concern. We can 

### 3 clearly state from our findings that no toxic substances are not 

### 4 a matter of concern. Regarding the question - do we need an 

### 5 extra test to assess the toxicity of dust compared to liquid 

### 6 formulations in a laboratory level? No, we can extrapolate from 

### 7 the dust toxicity from the feeding toxicity and normal toxicity. 

### 8 **DR. DANIEL SCHLENK:**I think this is just 

### 9 looking at exposure though, not necessarily toxicity. So, I 

### 10 guess the question - I could be wrong here, and other Panel 

### 11 members, but in terms of the Tier I exposure side is should dust 

### 12 be added into the other two variables. I guess that's how I see 

### 13 the question being asked. 

### 14 **DR. THOMAS POTTER:**I say yes. 

### 15 **DR. DANIEL SCHLENK:**That's what I thought. 

### 16 Yes? 

### 17 **DR. JEFF PETTIS:**If I understood you 

### 18 correctly that the difference between the dust application and 

### 19 oral or dermal, you could gain the same information by. 

### 20 **DR. DANIEL SCHLENK:**So the consensus is yes, 

### 21 or dust should be included. 

### 22 **DR. THOMAS POTTER:**Can I get in here, because 

### 23 now I'm confused. This is Tom Potter here. When we're talking 

### 24 about dust, are we talking about dust that was generated from 

### 25 treated seed? Are we talking about dust as a formulation of the 

### 26 pesticide? Or are we talking about the dust associated with

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1 wind-blown material from treated fields? So, I think we need to 

### 2 be clear in terms of what type of dust we are talking about in 

### 3 terms of exposure here. I'm a bit confused on that. 

### 4 **MR. JENS PISTORIUS:**Jens Pistorius. Thank 

### 5 you for this because I forgot to mention that it's not only seed 

### 6 treatments, which may generate such problematic dust. Also 

### 7 granules may. What we did is we did investigations with abraded 

### 8 dust. So basically, it's the formulation. 

### 9 **DR. DANIEL SCHLENK:**Yeah, Dr. Hunt? 

### 10 **DR. GREG HUNT:**I think there is some 

### 11 consensus. I think we're talking about two different things in 

### 12 a way, but I think there is consensus among the group that 

### 13 abraded dust from treated seed should be considered on 

### 14 off-target plants as another method of exposure and that we need 

### 15 to - and I think that's clear. Just looking at the 

### 16 concentrations that we see in bee collected pollen, I think that 

### 17 as my colleague, Christian Krupke mentioned, that this is an 

### 18 area where clearly there could be some mitigation, planters 

### 19 could be modified. We could, as Tom Potter mentioned, estimate 

### 20 how much dust is coming out of the planter. There could be 

### 21 changes in formulation. There could be other regulatory actions 

### 22 or reanalysis of the regulation of this using so much 

### 23 neonicotinoids over such a large acreage, especially since it 

### 24 has such a long half life than high water solubility and such 

### 25 high toxicity. 

### 26 **DR. DANIEL SCHLENK:**Okay. Any other

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1 comments? Dr. Pettis? 

### 2 **DR. JEFF PETTIS:**Well, I guess I would just 

### 3 like to hear maybe from the group or from EPA about EPA was 

### 4 proposing that mitigation measures might be the primary means of 

### 5 dealing with this dust issue. I don't know how the Panel feels 

### 6 about that, well other than - in other words, are mitigation 

### 7 measures at the time of planting sufficient for this issue? 

### 8 **DR. DANIEL SCHLENK:**Okay. 

### 9 **DR. THOMAS POTTER:**This is Tom Potter. Can I 

### 10 just comment then because I brought that up because honestly, I 

### 11 have no idea how effective the mitigation measures might be. I 

### 12 went and googled around it, but I didn't come up with anything. 

### 13 So, perhaps there's something marvelously effective? 

### 14 **DR. DANIEL SCHLENK:**Okay. Just, let's do one 

### 15 thing at a time. Is that something the Agency wishes to answer 

### 16 or has an answer for? 

### 17 **DR. STEVEN BRADBURY:**It's a challenging 

### 18 combination of entities that come to bear on reducing dust. The 

### 19 equipment manufacturers, the seed planters, and other 

### 20 manufacturers, the pneumatics - I'm not an engineer, I don't 

### 21 pretend to be one, but you all get that part. Then there's the 

### 22 pesticide itself and the product in the sticking agent that the 

### 23 registrant, but also the seed company are in play there. Then 

### 24 there's talc or graphite or other components that are mixed in 

### 25 with the seeds so they don't in the planter. The seed treatment 

### 26 is a little sticky because it wants to hold the fungicide and

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1 insecticide on. 

### 2 So without getting too much into statutory 

### 3 stuff, the FIFRA label only goes so far in terms of what we can 

### 4 or can't do in terms of asking for data. Then there are things 

### 5 we do that aren't on the label, but part of the stakeholders 

### 6 getting together and trying to work through some solutions. 

### 7 There was a public meeting mid July, end of July at Iowa State 

### 8 and the State of Iowa convened in sort of an open meeting to 

### 9 sort of brain storm on this very topic. Some companies were 

### 10 describing with how they're coming up with some alternatives to 

### 11 talc, and they're working with the seed planting companies and 

### 12 they're working with registrants and they're working with 

### 13 formulators. In fact, there is equipment that can be used to 

### 14 measure how much dust is coming off the benchtop in the 

### 15 laboratory, sort of reasonable surrogates to get a sense of 

### 16 different combinations of material can change the amount of 

### 17 abrasion that occurs, and you can measure the amount of the 

### 18 pesticide in that dust. 

### 19 So, as far as data, it appears that there are 

### 20 people working on the kinds of experiments where you could start 

### 21 to get some insight, which then could give us some ways of 

### 22 between what you could potentially do. I'm not trying to 

### 23 project too far into the future, but theoretically some things 

### 24 you might be able to do on a FIFRA label versus things that play 

### 25 out in best management practices, things USDA and others could 

### 26 be doing in terms of improving DMPs, which may be related to

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1 dust management in general. There is probably going to be a 

### 2 combination of data we can get and some experiments to get 

### 3 insights into how products are used. 

### 4 **DR. DANIEL SCHLENK:**Okay. Does that answer 

### 5 your question Tom? 

### 6 **DR. THOMAS POTTER:**Yes, I think very well. 

### 7 Obviously it is in our complex set of characters involved. If I 

### 8 could just offer one more comment on this, I think that by 

### 9 including some estimate of exposure associated with dust 

### 10 unmitigated, it may in some sense motivate, force, compel other 

### 11 parties to you know, increase their effort in this mitigation 

### 12 area. 

### 13 **DR. STEVEN BRADBURY:**I would like to say one 

### 14 thing that information that was shared at this meeting at Iowa 

### 15 State wasn't the worst bias to the credit of the seed planting, 

### 16 the John Deere and the other companies and folks working on 

### 17 formulations and some registrants. They were working on it on 

### 18 their own. So, I think people are working hard and trying to 

### 19 figure out the engineering and the chemistry and the efficacy of 

### 20 the material for plant protection. And there were beekeepers at 

### 21 this meeting. It was quite a good meeting actually, people 

### 22 trying to come up with solutions. 

### 23 **DR. DANIEL SCHLENK:**That was Dr. Bradbury. I 

### 24 think we had Dr. James first? You put your hand back down. 

### 25 **DR. ROSALIND JAMES:**I just wanted to 

### 26 reiterate, I guess, about the dust from pesticide applications

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1 that are not soil treatments also, and make sure that doesn't 

### 2 get lost in this discussion. 

### 3 **DR. DANIEL SCHLENK:**And Dr. Berenbaum? 

### 4 **DR. MAY BERENBAUM:**I just was not clear on 

### 5 whether EPA has regulatory enforcement authority over dust? I 

### 6 mean, is that within bailiwick or. 

### 7 **DR. STEVEN BRADBURY:**Not under FIFRA. If 

### 8 there is a formulation that is a dust, then that is covered. If 

### 9 it gets a little less - if it's dust blowing off of soil from 

### 10 soil on a field that has been in corn for two to three to four 

### 11 years, and there is probably something in there along with the 

### 12 nitrogen and phosphorus. That blowing around - FIFRA is not 

### 13 capturing that. It could be aspects of the Clean Air Act that 

### 14 are capturing that. You have probably heard of the PM10 rule 

### 15 and there is a PM10 rule now in the agricultural setting that is 

### 16 working through. So, without projecting too far, there is 

### 17 probably some air mottling that Office of Air did try to 

### 18 understand PM10 movement. There may be a way to tap into some 

### 19 of that at the bounding estimates, but you start to get the 

### 20 shade of gray in terms of where one statute drops off and 

### 21 another one starts. 

### 22 **DR. DANIEL SCHLENK:**That was Steve Bradbury. 

### 23 Dr. Hunt? 

### 24 **DR. GREG HUNT:**I guess I'm a little out of my 

### 25 element here because I'm not a toxicologist. I'm just a bee 

### 26 researcher, but I guess we're just looking at the risk

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1 assessment side. We're not looking at the benefit side. To me, 

### 2 that is an unknown because we don't have access to these yield 

### 3 studies. I don't know if they've been done just with 

### 4 neonicotinoids or with the entire seed treatment mixture, but it 

### 5 seems like this already registered product is being used on a 

### 6 wide scale. It's kinda of an experiment ongoing. I've talked 

### 7 to the chemical companies. They are working on litigation and 

### 8 they've shown me data, how different seed treatments come off 

### 9 the seeds more than others. Corn is the worst in that regard. 

### 10 I guess I'm wondering what is the EPA thinking in the big 

### 11 picture with neonicotinoid seed treatments. I don't know about 

### 12 regulatory laws very much. Is there any consideration. 

### 13 **DR. DANIEL SCHLENK:**Dr. Hunt, that's getting 

### 14 a little bit into policy and sort of away from the question that 

### 15 we're being asked at this particular point. Let me try to bring 

### 16 us back to that question, which is basically should dust be 

### 17 included in the Tier I assessment and what other ways do we 

### 18 quantify that. From what I understand is that we are in 

### 19 consensus that dust should be used. Apparently, there have been 

### 20 several recommendations of different methods to do that. Is 

### 21 there anything else that we need to add on that? Yes, Mr. 

### 22 Pistorius? 

### 23 **MR. JENS PISTORIUS:**Jens Pistorius. I think 

### 24 dust is not only an issue for maze, maybe also for (inaudible), 

### 25 but there is large number of other crops where dust is not an 

### 26 issue at all because of the seed treatment quality, which is

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1 very different for different seeds. 

### 2 **DR. DANIEL SCHLENK:**If you can, again, give 

### 3 those comments to Dr. Potter to simulate that information, that 

### 4 would be great. Dr. Hunt? 

### 5 **DR. GREG HUNT:**I'm sorry, but the question 

### 6 actually was should dust be considered as a route of exposure, 

### 7 and I think we addressed that. 

### 8 **DR. DANIEL SCHLENK:**Yes, that's what I said, 

### 9 yep. Okay. Any other comments on letter B? Okay. Let me go 

### 10 to Ms. Wendel. Are you clear with Panel's answer? Question or 

### 11 clarification? 

### 12 **MR. KEITH SAPPINGTON:**Clarifying question - 

### 13 Keith Sappington. If we would be interested in the methods of 

### 14 how you would incorporate a dust estimate of exposure a priority 

### 15 in the Tier I assessment. 

### 16 **DR. DANIEL SCHLENK:**Okay. I think we are 

### 17 clear on that. Okay. Let's move onto C, and read that into the 

### 18 record please. 

### 19 **MS. CHRISTINA WENDEL:**Christina Wendel, 

### 20 question number 7, subpart (c), drinking water -- please comment 

### 21 on the analysis, discussions provided in Appendix 2 of the White 

### 22 Paper and the conclusion that pesticide exposure to bees through 

### 23 drinking contaminated water is not expected to be a major route 

### 24 of exposure when compared to contact following foliar spray 

### 25 applications and diet. If the Panel believes that this exposure 

### 26 route should be quantitatively included in the Tier I exposure

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1 method, for the relevant application types, i.e. foliar spray, 

### 2 soil treatment, seed treatment, or trunk spray, please discuss 

### 3 why and what data may be needed to address the exposure route 

### 4 quantitatively. 

### 5 **DR. DANIEL SCHLENK:**And Dr. Potter. 

### 6 **DR. THOMAS POTTER:**Okay. Well, I think it 

### 7 was said earlier that we felt like it was important for this 

### 8 route of exposure to be included. I believe that Tier I, we 

### 9 haven't had a discussion among the group here. So, we may 

### 10 disagree, but I don't think we will. So, the short answer is 

### 11 yes, it should be included. 

### 12 Okay. Now the second part of the answer, 

### 13 we're going to need to look at two different issues that 

### 14 ultimately translate to exposure. One is potential 

### 15 concentrations of pesticides in water that bees may be exposed 

### 16 to. That was addressed to some degree in the appendix, and 

### 17 we'll get to that, but I just wanted to mention the second part, 

### 18 and that is the question of just how much water do bees and/or 

### 19 other pollinating organisms consume on a daily basis. Clearly, 

### 20 we need to multiply the two amount times concentration to come 

### 21 up with exposure. So, we will break out the answer into those 

### 22 two parts in terms of the comment. 

### 23 First with regard to potential concentrations 

### 24 of active ingredients in water sources that bees may use, I 

### 25 believe and I think there will be consensus on that, that the 

### 26 concentration levels that we generally find in surface waters in

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1 aquacultural and in urban areas across the United States are 

### 2 generally very low. Typical datasets that are available were in 

### 3 the parts per billion or subparts per billion level for most 

### 4 active ingredients. So, these are exceedingly low relative to 

### 5 potential exposures given estimated volumes of water consumed 

### 6 during the day. I would say, generally data that is out there 

### 7 supports the conclusion that water should not be concluded. So, 

### 8 there, I've backed up and gone in the opposite direction here, 

### 9 but we'll get there. 

### 10 The second part of this is that the Agency in 

### 11 their appendix went through and did a couple of assessment of 

### 12 potential exposures. One of them, they used the model GENEEC, 

### 13 which is the standard model to use, screening model to be used 

### 14 to estimate pesticide exposures in drinking water and surface 

### 15 water. It's a first tier model. I believe that model is used 

### 16 correctly. Basically, it shows that even in a worst case 

### 17 scenario, if you are looking at a farm pond, which has a lot of 

### 18 water in it, obviously - I mean, I'm not saying that 

### 19 facetiously. So, there is a potential for substantial dilution. 

### 20 The concentrations again are relatively low. So, that's not 

### 21 likely, again, to be a significant exposure pathway. 

### 22 Finally, the third area that we may want to 

### 23 consider in this, is the idea of puddles that are left in the 

### 24 field, either from irrigation water or rain. Those can be 

### 25 directly sprayed as noted in the appendix. In their 

### 26 deliberations, Agency staff went through and did some

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1 calculations using a simple equilibrium model that I think was 

### 2 extracted from a rice paddy assessment that was done regarding 

### 3 pesticides the use in rice paddies. I went in and I looked at 

### 4 that. I think that there was an error in the calculations. I 

### 5 will provide this in written comment. That error resulted in 

### 6 the estimated concentrations in puddles being essentially 1000 

### 7 times lower than what they would likely be using the same set of 

### 8 assumptions. So, anyway, I think that ends up indicating that 

### 9 puddles that are in fields that are sprayed may have relatively 

### 10 high concentrations of pesticides. We're talking about in the 

### 11 parts per million level. If we use those values and then go 

### 12 into looking at the volume of water that bees may consume on a 

### 13 daily basis. 

### 14 First, if we use the low value, which was 

### 15 recommended, I think, from the paper wasp study -- is that 

### 16 correct? Forty-five microliters a day, if we multiply that 

### 17 times these estimated high-end estimated concentrations in 

### 18 pesticide contaminated puddles, the drinking water exposure 

### 19 could be somewhere in the order of 10 percent to 20 percent of 

### 20 the dietary exposure. So, I think that's a potentially 

### 21 significant exposure pathway and should be included. 

### 22 Further, if we go to the second part of the 

### 23 exposure equation, which is that the volume of water consumed, I 

### 24 certainly would like to hear from some of the other panel 

### 25 members who are more knowledgeable about bees than I am. It 

### 26 struck me that there was a substantial uncertainty with regard

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1 to the volumes that may be consumed on a daily basis. The 

### 2 assessments that were made indicated they varied from somewhere 

### 3 in the order of 45 microliters a day to 6 milliliters a day, 

### 4 which is a factor of about 100. 

### 5 So, if we do some bounding estimates here 

### 6 using those high-end concentrations that we could estimate by 

### 7 doing our equilibrium model for pesticide-contaminated puddles. 

### 8 Drinking water exposure could indeed exceed dietary exposure. 

### 9 So therefore, I strongly feel that it needs to be captured in a 

### 10 Tier I assessment, and perhaps some effort could also be made in 

### 11 terms of further refining potential environmental 

### 12 concentrations. I made this comment yesterday in the form of a 

### 13 question. But in ECOFRAM, there is a module that actually 

### 14 looked at runoff in fields and accumulation in puddles using 

### 15 some subcomponents of PRZM, relatively simple model to use and 

### 16 run. Perhaps that should be an alternate approach in terms of 

### 17 estimating pesticide contamination in field puddles that might 

### 18 provide some insight into this exposure issue. 

### 19 **DR. DANIEL SCHLENK:**Thank you, Dr. Potter. 

### 20 Dr. Hunt? 

### 21 **DR. GREG HUNT:**I'm not sure, but I believe 

### 22 that bees get most of the dietary water through nectar. I would 

### 23 need to look in the literature to confirm this. They will 

### 24 collect water in the fields or in puddles primarily to cool the 

### 25 hive, the deposit in the comb, to cool the hive and also to 

### 26 dilute the brood food, but they will also use nectar for that.

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1 That's my understanding. 

### 2 **DR. DANIEL SCHLENK:**So, I guess you would say 

### 3 the water should not be included - is that. 

### 4 **DR. GREG HUNT:**I'm saying I don't know. 

### 5 **DR. DANIEL SCHLENK:**Okay. 

### 6 **DR. THOMAS POTTER:**Can I ask a followup on 

### 7 that? 

### 8 **DR. DANIEL SCHLENK:**Sure, yeah. 

### 9 **DR. THOMAS POTTER:**This is Tom Potter here. 

### 10 I think what we're talking about is around the lunch break, and 

### 11 I was asking how do the bees get the water back to the hive? 

### 12 What my understanding was that they were essentially ingesting 

### 13 it and then regurgitating it. 

### 14 **DR. GREG HUNT:**Yes, that's right. They 

### 15 ingest it into the honey stomach, which is like a waterproof bag 

### 16 more or less, but they have the pyloric valve from the honey 

### 17 stomach to the midgut and some water could pass through that. 

### 18 And I suppose pesticides could be absorbed through the walls of 

### 19 the stomach. 

### 20 **DR. DANIEL SCHLENK:**Okay. Dr. Pettis? 

### 21 **DR. JEFF PETTIS:**I'll just a bit to what Dr. 

### 22 Hunt said about water used by bees. Primarily for two reasons, 

### 23 as you said. One is to dilute honey to then use as food and 

### 24 also to dilut brood food somewhat. The big issue - so the most 

### 25 volume is in cooling the hive. There again, certain bees go 

### 26 out, they collect the water and bring it back in. They can

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1 actually sit in the hive and hold his water in their body for 

### 2 extended periods of time, act like little water tanks held 

### 3 within. Then, they move it around the hive and evaporate it so 

### 4 it can be put into cells and evaporate in the cells. So, at 

### 5 certain times, and in climatic conditions where you have hot 

### 6 weather, the use of water can be extensive. 

### 7 I would probably agree with you Greg, that 

### 8 that in general under non-hot conditions that there is a lot of 

### 9 - the water requirements are probably met by consuming honey 

### 10 and/or nectar. But when you have hot conditions, certainly the 

### 11 increase for cooling needs is great. So, I think from my 

### 12 perspective, I would think that it can't be discounted as a 

### 13 source of exposure. How will you incorporate it into the Tier I 

### 14 - I'm uncertain at this time. 

### 15 **DR. DANIEL SCHLENK:**Mr. Pistorius and then 

### 16 Dr. Fefferman? 

### 17 **MR. JENS PISTORIUS:**I have a proposal how to 

### 18 integrate it in Tier I risk assessment, and it will be published 

### 19 in about one or two weeks along with the run of data on 

### 20 guttation issues published in the proceedings of the ICPBR. 

### 21 Basically, what we've done in Germany again, is a lot of 

### 22 different crops. We looked at the residue that get into the 

### 23 guttation droplets and then also, we did semi-field and field 

### 24 trials with worst case and realistic exposure. What you can say 

### 25 - okay in semi-field, you get high mortality if you don't give 

### 26 any other water to the bees, but if you give another water

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1 source, it is quite lower. 

### 2 Now to the field test and monitoring that we 

### 3 have done, we have had, not only by us but agricultural estates 

### 4 and institutes - about 12 different monitoring with a lot of bee 

### 5 hives and worst case exposure directly at maze fields. 

### 6 In one of the trials, we had on two days 

### 7 increased mortality. So, as a general answer to the question 

### 8 that is asked here - what was it? Drinking guttation water is, 

### 9 in my opinion, not expected to be a major route of exposure when 

### 10 compared to contact and diet is what I agree upon. I'm not 

### 11 quite sure what it is like here for all the desert situations. 

### 12 That is maybe different. I can only say from my findings. What 

### 13 we can do is have a little bit of distance, so it is a 

### 14 mitigation measure between highest and the crop, whoever has to 

### 15 keep this distance. 

### 16 **DR. DANIEL SCHLENK:**Dr. Fefferman? 

### 17 **DR. NINA FEFFERMAN:**So, while I don't know to 

### 18 speak to the drinking water per se, I agree that most of the 

### 19 water collected is not being used for drinking. I think that 

### 20 same intent - although maybe this comes under other exposure 

### 21 routes and I don't know, its still water. By virtue of the fact 

### 22 that that water is being collected for cooling, I think plays 

### 23 directly into what we were talking about with the answer to the 

### 24 early question about wax exposure. 

### 25 As it's circulated around the wax and 

### 26 evaporated, I don't know how much pesticide evaporates with that

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1 water and how much is left residually on the wax. I think that 

### 2 actually might be, especially at the hottest points in the 

### 3 summer, a substantial introduction of pesticide into the wax 

### 4 comb itself and that may be a temporal aspect of all of this 

### 5 that might be important to look into. 

### 6 **DR. DANIEL SCHLENK:**It sounds like a seasonal 

### 7 component for sure. Any other comments? Yep, Dr. Tarpy? 

### 8 **DR. DAVID TARPY:**I just wanted to reiterate 

### 9 that point too, is that if using mostly for cooling, the three 

### 10 studies that are cited in Appendix 2, at least one of them is 

### 11 really done in upstate New York, right? Where it doesn't get 

### 12 above 70 degrees in July, right? So, in other places, Central 

### 13 Valley of California, with 120 degrees and the bees keeping the 

### 14 brood nests at 95 degrees, it takes a substantial amount of 

### 15 water for that cooling to occur. So, I would say that those 

### 16 estimates in worst case scenarios could be incredibly low, but 

### 17 there seems to be a data gap there obviously and very high water 

### 18 needs, not drinking water, but high-cooling water needs. 

### 19 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 20 **DR. MAY BERENBAUM:**Shame on me for 

### 21 introducing anecdotal unscientific observations here. I'm 

### 22 asking my bee keeping colony - I'm not a bee biologist, but I'm 

### 23 the one who gets the calls from people who want to save the bees 

### 24 and help the bees. I can't tell you how many calls I've gotten 

### 25 from people who have had bees that drown in bird water baths - 

### 26 that's the word. So, my impression is that a tiny bit

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1 unscientific, I can't site literature here, but bees do visit 

### 2 water and get immersed in water. So, that's totally anecdotal, 

### 3 but that may be something to consider, particularly because 

### 4 there can be puddles that vary in their pesticide content. 

### 5 That's not unexpected. I don't know how that fits into 

### 6 calculations of exposure. 

### 7 **DR. DANIEL SCHLENK:**Yes, Dr. Pettis? 

### 8 **DR. JEFF PETTIS:**We would agree with you that 

### 9 bees do visit water and they need water. We were asked to 

### 10 comment on the exposure method relevant to the application 

### 11 types, foliar spray, soil treatments, seed treatments, trunk 

### 12 spray. There might be an important component here for the type 

### 13 of chemical involved, whether it is water soluble or insoluble 

### 14 in water. I would think that maybe the agency could take into 

### 15 account - if it's insoluble in water, then only foliar 

### 16 applications may be an issue were it collects in puddles or 

### 17 temporary things, whereas water soluble compounds, soil 

### 18 treatments, seed treatments, et cetera, could move through the 

### 19 soil and accumulate in water. So, there could be, based on the 

### 20 chemical type, we may consider that. Some of the studies 

### 21 looking at residues in standing water or temporary water, 

### 22 they've looked at Imidacloprid, which is water soluble. 

### 23 **DR. DANIEL SCHLENK:**Yes, Mr. Pistorius? 

### 24 **MR. JENS PISTORIUS:**I completely agree. It's 

### 25 no problem for a nonsystemic and only for highly toxic - it may 

### 26 only be a potential problem for highly toxic substances.

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1 **DR. DANIEL SCHLENK:**Okay. Anyone else? 

### 2 Sounds like you got a range of answers there. Sounds like 

### 3 definitely maybe on that one. So, we will leave that to Dr. 

### 4 Potter to put all of those answers together in some coherent 

### 5 fashion there. You've heard several answers here. Do you have 

### 6 any questions or clarification? 

### 7 **MR. KEITH SAPPINGTON:**Keith Sappington. 

### 8 Yeah, just methods for quantifying it, but yes, drinking maybe. 

### 9 10 **DR. DANIEL SCHLENK:**Okay. Is that clear to 

### 11 you? 

### 12 **DR. THOMAS POTTER:**Tom Potter here. I just 

### 13 want to say, you know, and thanks for all the discussion here, I 

### 14 think this has been very informative, certainly to me, but it's 

### 15 not just drinking water. Obviously, it is water. And so, we 

### 16 ought to refine our terminology here to say water used. 

### 17 **DR. DANIEL SCHLENK:**Sure. Good point. Okay. 

### 18 Let's move onto - I think this might be our last one of the day 

### 19 here, perhaps, letter D. 

### 20 **MS. CHRISTINA WENDEL:**Christina Wendel. 

### 21 Question 7, subpart (d), other routes - please identify and 

### 22 discuss additional exposure routes, if there are any besides 

### 23 contact with dust and consumption of drinking water, that would 

### 24 contribute significantly to pesticide exposure of bees and 

### 25 explain how and why such exposures could be considered 

### 26 quantitatively in establishing the Tier I exposure value.

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1 **DR. DANIEL SCHLENK:**Dr. Potter? 

### 2 **DR. THOMAS POTTER:**Well, I think I'm going to 

### 3 send this over the Rosalind - just teasing, but hopefully you'll 

### 4 comment on this. I feel that the route of exposure that would 

### 5 need to be considered here would be direct soil contact. I 

### 6 think Rosalind described earlier that there are many bees who 

### 7 come in directly in contact with soil using the material for 

### 8 nesting, nest building and et cetera. So certainly, there is 

### 9 potential for contact. Now the question of how we would 

### 10 evaluate that quantitatively, I'm hoping somebody here is going 

### 11 to help me with that. 

### 12 **DR. DANIEL SCHLENK:**Dr. Hunt, any help for 

### 13 Dr. Potter there? 

### 14 **DR. GREG HUNT:**Not really. One other obvious 

### 15 route of exposure that we haven't talked a lot about is the wax 

### 16 comb itself with the brood swimming in royal jelly, which is 

### 17 liquifilic and the wax comb, which is liquifilic, but that would 

### 18 come under food, I mean ingestion or absorption through the 

### 19 cuticle. But, I don't think that needs to be, in my opinion, 

### 20 considered under Tier I exposure, but it's just another route of 

### 21 exposure. 

### 22 **DR. DANIEL SCHLENK:**So, not a significant 

### 23 route of exposure, not to be included in that first tier 

### 24 approach. Okay. And Dr. Pettis? 

### 25 **DR. JEFF PETTIS:**Really, not much to add that 

### 26 hasn't been already said, other than the fact that bees are

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1 great samplers of the environment, and they have these hairs 

### 2 that will pick up all kinds of dust and particles and whatever, 

### 3 but it's not really - it's more of a general issue with bees, 

### 4 not so much a regulatory issue. 

### 5 **DR. DANIEL SCHLENK:**Well, what do you say, 

### 6 Dr. James - are you going to help Dr. Potter out there? 

### 7 **DR. ROSALIND JAMES:**I'm probably just 

### 8 reiterated what I have said earlier today in that if you are 

### 9 going to use honey bees as a surrogate for other bees, you need 

### 10 to think about where other bees go besides just where honey bees 

### 11 go, and you should add contact toxicity for soil pesticides. 

### 12 So, you already have some estimates and some methods for 

### 13 estimating what concentration of pesticides should be in the 

### 14 soil, and I recommend adding larval contact toxicity test for 

### 15 pesticides that are in the soil, and the reasoning being you 

### 16 have your soil nesting bees and you also have mason bees that 

### 17 are using mud for agricultural fields. 

### 18 **DR. DANIEL SCHLENK:**Okay. Anyone else? Oh 

### 19 sorry, Dr. Hunt? 

### 20 **DR. GREG HUNT:**Just a question for Rosalind. 

### 21 So, that would be kind of like the brood food. I mean, if you 

### 22 spike the brood food, then there are contacts with that, right? 

### 23 Would that cover it? 

### 24 **DR. ROSALIND JAMES:**But this has to do with 

### 25 exposures and your estimating exposure levels from soil, I guess 

### 26 is the difference. What is a contact exposure level from soil?

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1 2 **DR. DANIEL SCHLENK:**Okay. Any other comments 

### 3 for question 7? Going once, going twice. Okay. Let me go to 

### 4 the Agency. Are you clear or have any questions or 

### 5 clarification in terms of the Panel's response? We're good to 

### 6 go. Okay. Well, let's call it here for a break until tomorrow. 

### 7 We've made great headway. Congratulations to the Panel and 

### 8 appreciated for your hard work and meeting together in getting 

### 9 these things efficiently done without redundancy. We are about 

### 10 one-third of the way done, a bit over a third of the way in 

### 11 terms of number of questions, although there are 14 total 

### 12 questions - there's seven, but there are a lot of other subparts 

### 13 that we have to go through yet. So with that, I will turn it 

### 14 over to Fred for some administrative comments. 

### 15 **DR. FRED JENKINS:**I don't have a whole lot 

### 16 extra to add, but I just want to thank the panel for a good day 

### 17 of deliberations and also the public commenters. I think we 

### 18 accomplished a lot today. So, I just wanted to say to the 

### 19 panel, remind them to please return your thumb drives to us with 

### 20 the presentation, so we can have those recycled. And with that 

### 21 said, we will see everyone in the morning at 9 a.m. Have a 

### 22 great evening. 

### 23 (WHEREUPON the meeting was adjourned for the 

### 24 day.) 

### 25 **DR. DANIEL SCHLENK:**Good morning everyone. I 

### 26 think in lieu of time, we are going to sort of skip our

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1 introductions since, I think everybody know everyone by now, I 

### 2 hope. This morning, the Agency is going to hit us with some 

### 3 questions of clarification, I believe, on question 7, is that 

### 4 correct? Yep, anybody who is going to be doing that. Okay. 

### 5 **MR. KEITH SAPPINGTON:**Thank you, Dr. Schlenk. 

### 6 I would like to offer, on behalf of the Agency, just a little 

### 7 question of clarification on charge question 7, but first, we 

### 8 want to thank again the Panel for all their work over in 

### 9 preparing for this meeting and the deliberations of the last 

### 10 couple of days. We've already gained quite a lot of insight on 

### 11 how we can improve the proposed process. So, we really 

### 12 appreciate that. 

### 13 With regards to question 7, and I know the 

### 14 Panel has all ready considered this, so this is perhaps more of 

### 15 an emphasis and that is in considering the alternate pathways in 

### 16 Tier I exposure, it would be helpful to us in understanding the 

### 17 conditions in which considering these additional pathways, like 

### 18 drinking water and dust in associated oral exposure from dust 

### 19 onto other plants, conditions in which that might change the 

### 20 outcome of the Tier I assessment from say passing the assessment 

### 21 to not passing the assessment since our goal is to minimize type 

### 22 2 errors. 

### 23 So, in thinking about that and then in 

### 24 thinking about the different context, so perhaps for example, 

### 25 for the seed treatment where we have a default value of 1 mg per 

### 26 kilogram. Is there is enough margin and safety such that not

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1 including drinking water or some of these other sources, would 

### 2 that likely really push that from a no risk or no LOC exceedance 

### 3 to a yes exceedance. Thank you very much. 

### 4 **DR. DANIEL SCHLENK:**I believe that Dr. Potter 

### 5 is the lead on question 7, so do you have any questions related 

### 6 to how you want to present that? 

### 7 **DR. THOMAS POTTER:**Well, I have a comment. I 

### 8 probably have to come up with a few questions too here. Anyway, 

### 9 good morning, Tom Potter here. Are you specifically asking 

### 10 about the seed treatment scenario only or all of the scenarios 

### 11 that we were looking at? 

### 12 **MR. KEITH SAPPINGTON:**Keith Sappington. All 

### 13 the scenarios and considering the alternate routes. 

### 14 **DR. THOMAS POTTER:**Right. You know, one of 

### 15 the things that, you know, in terms of, let's just break them 

### 16 out here. I'll allow for a few thoughts on it. I saw Dr. Hunt 

### 17 also had a comment that he wanted to make, but you know one of 

### 18 the things about the dust is that hopefully there will be some 

### 19 effort to look at the potential exposures. Perhaps you've done 

### 20 that all ready. So, if indeed you could come to some well 

### 21 established conclusion that that is not an issue, it certainly 

### 22 may not belong in Tier I. This is my opinion. 

### 23 So there needs to be some assessment, 

### 24 particularly in regard to wind-blow soil dust. That's kind of 

### 25 my issues. We will let Jens talk about the abraded material 

### 26 from seed coat in a few minutes perhaps.

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1 With regard to drinking water, I would offer 

### 2 the same line of thought or reasoning. I think that, you know, 

### 3 a reassessment to determine if there are potentially significant 

### 4 exposures. I think we noted yesterday that there was 

### 5 considerable uncertainty in the amount of water that bees drink 

### 6 and come in contact with on a daily basis, varying on a factor 

### 7 of a range of 100. So, certainly that could have a major impact 

### 8 on potential exposures. 

### 9 And then there was the question of assessing 

### 10 potential concentrations, in again a worst case scenario and 

### 11 overspray on a field that had standing water in it, using the 

### 12 type of exposure scenario to come up and say well, you know is 

### 13 this 1 percent of the dietary contact exposure or is it 50 

### 14 percent or is it greater than the dietary - I mean if you use 

### 15 the higher numbers for the water exposure in high end numbers 

### 16 for field puddles, you can come up with a pretty significant 

### 17 load. No, I don't necessarily think that that's reasonable or 

### 18 realistic thing to do, but I think it's something that you 

### 19 should spend some more time working on in the process of 

### 20 refining the White Paper. 

### 21 **DR. DANIEL SCHLENK:**Dr. Hunt? 

### 22 **DR. GREGORY HUNT:**Well, in considering, for 

### 23 example, the abraded seed coat dust, I don't think that 

### 24 considering that route of exposure would affect your Tier I, you 

### 25 know, because 1 mg per kilogram is very conservative. It might 

### 26 affect the downstream, for example, and I look at the figures on

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1 the tiered assessment. Downstream if it exceeds the LOC, then 

### 2 it says use available crop residue studies, recalculate our 

### 3 cues. So, in that blight, the crop residue studies may be 

### 4 should include like incoming pollen that the bees are bringing 

### 5 in. What are the maximal values you are seeing in those? 

### 6 **DR. DANIEL SCHLENK:**Dr. Pettis, you're also 

### 7 on this one too. 

### 8 **DR. JEFF PETTIS:**I have no immediate comments 

### 9 at this time. 

### 10 **DR. DANIEL SCHLENK:**Dr. Fefferman, and then. 

### 11 **DR. NINA FEFFERMAN:**Thanks. I think one 

### 12 major thing I took away from the conversation yesterday from 7 

### 13 is maybe even the concept of the phrase drinking water might be 

### 14 an inappropriate way to frame that, and that just water usage 

### 15 might be a way to frame that. While I don't know, again, 

### 16 whether or not on testing this will turn out to be significant. 

### 17 It does lead, in my mind, to the suggested additional outcome 

### 18 that maybe wax sampling concentrations should be measured as 

### 19 part of a Tier I assessment, because that could tell us a 

### 20 gradual accumulation over time as opposed to a dose response 

### 21 from an initial carrying of what exposed larvae are facing or 

### 22 what recycling of wax within the hive is doing. 

### 23 So, if we're worried about deposition 

### 24 gradually over time from evaporation leaving not so much 

### 25 drinking water but evaporated water for cooling leaving residues 

### 26 of pesticides within the wax. Then that suggests to me not only

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1 a potential for it to trigger a positive case where maybe it 

### 2 should be. Not only false but true, but also a new mechanism to 

### 3 measure, which is the concentration in the wax. 

### 4 **DR. DANIEL SCHLENK:**Okay. Mr. Pistorius? 

### 5 **MR. JENS PISTORIUS:**I want to thank EPA for 

### 6 this excellent question. No, it will not change the outcome of 

### 7 the risk assessment and the Tier I seems to be conservatively 

### 8 enough that the substances of concern are identified. So, it is 

### 9 an issue of higher tier testing to address if there are concerns 

### 10 under realistic conditions, but it will not change. Also the 

### 11 route of dust and guttation, and I think also for the risk of 

### 12 water, a lot changed the outcome of the risk assessment. You 

### 13 will be able to identify the substances of concern in Tier I. 

### 14 **DR. DANIEL SCHLENK:**Okay. And Dr. James, did 

### 15 you have your. 

### 16 **DR. ROSALIND JAMES:**Wouldn't it change a risk 

### 17 assessment for nonsystemic pesticides applied to the soil with 

### 18 the dust because the dust would provide another route of 

### 19 possible exposure that you didn't account for with a 

### 20 nonsystemic? You talk a lot about nonsystemic soil-applied 

### 21 pesticides. 

### 22 **DR. DANIEL SCHLENK:**Mr. Pistorius? 

### 23 **MR. JENS PISTORIUS:**I think it is both for 

### 24 the systemic and nonsystemic that the Tier I level will be able 

### 25 to identify those substances of concern where you find a high 

### 26 toxicity and you have a classification as highly toxic

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1 substance, which will detect in the laboratory, oral and contact 

### 2 toxicity tests. You will be able to identify substances with 

### 3 potential concern. For nonsystemic substances, guttation is not 

### 4 an issue at all, but I agree dust is for highly toxic 

### 5 substances. 

### 6 **DR. ROSALIND JAMES:**Rosalind James. But I'm 

### 7 out for dust, not guttation. Right. 

### 8 **MR. JENS PISTORIUS:**Yes, as I said for dust, 

### 9 highly toxic substances are a concern no matter if systemic or 

### 10 nonsystemic. 

### 11 **DR. DANIEL SCHLENK:**Anyone else want to weigh 

### 12 in on this one? Okay. Mr. Sappington, does that answer 

### 13 questions? Are you guys good with that? 

### 14 **MR. KEITH SAPPINGTON:**Thank you very much. 

### 15 It helps a lot. 

### 16 **DR. DANIEL SCHLENK:**Okay. Great. Okay. At 

### 17 this point in time, I guess we are ready to move onto question 

### 18 8, and if you could read in letter A for question 8? 

### 19 **MR. JOSEPH DECANT:**Good morning. This is Joe 

### 20 Decant of EFED. So, I'll start us off on the toxicity side. 

### 21 Question 8, subpart (a) - please comment on the extent to which 

### 22 currently available bee toxicity tests, which focus primarily on 

### 23 mortality or survival, serve as an effective Tier I screen. 

### 24 **DR. DANIEL SCHLENK:**And our lead discussant 

### 25 is David Tarpy. 

### 26 **DR. DAVID TARPY:**Thank you, Mr. Chairman.

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1 Good morning everybody. This is David Tarpy with North Carolina 

### 2 State University. I just want to thank the EPA and those who 

### 3 crafted the White Paper are really doing an excellent job. It's 

### 4 been incredibly informative to me. It's been obviously very 

### 5 thorough, very well articulated, and I think very helpful to 

### 6 have this discussion. I very much welcome it and I'm thrilled 

### 7 we're all doing it. 

### 8 I foresee our conversations today looking at 

### 9 the effects on honey bees as a proxy for all bees and 

### 10 pollinators as being very important because the use of a social 

### 11 system to study solitary systems in general, it adds many 

### 12 complexities of biological organization that need to be taken 

### 13 into account, particularly at Tier I types of bioassays. So, I 

### 14 think we have a lot of thoughts concerning that that we'll be 

### 15 discussing today. 

### 16 A couple of points that the discussants for 

### 17 this particular question wanted to bring up before we get into 

### 18 other details and later sub-questions and later question, one is 

### 19 to reiterate the issue of using the individual bee as the basal 

### 20 determinator rather than standardizing for body weight of the 

### 21 individual, especially if honey bees are going to be used as a 

### 22 proxy for other types of bees that can vary quite dramatically 

### 23 in their size. Seemingly to standardize on a milligram per bee 

### 24 is going to be much more translatable to those other symptoms 

### 25 than just doing it on a per bee basis. 

### 26 Also, because honey bees are obligatorily

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1 social as the White Paper very well articulates, they're a super 

### 2 organism, and that introduces a lot of important biological 

### 3 phenomenon to measuring them in vitro and in vivo. Sometimes 

### 4 those can be tested - sometimes honey bees can be tested in a 

### 5 vacuum as an individual, but often times, it's not very 

### 6 biologically meaningful. So, including the social context in 

### 7 Tier I screenings I think is going to become very important. 

### 8 Another aspect, and this derives from the 

### 9 social complexity of honey bees as social insects is that a lot 

### 10 the Tier I screening techniques that's commonly used for 

### 11 solitary insects don't really capture a temporal component to 

### 12 the potential effects of screening for these compounds. Cutting 

### 13 it off at that 48 hours or extending that to 72 or even 96 hours 

### 14 doesn't really capture a lot of the social effects over time. 

### 15 So, adding that additional dimension to these bioassays we think 

### 16 would be very informative, and we'll be discussing those in a 

### 17 little more detail. 

### 18 Specifically to address the need for measuring 

### 19 outcomes of individual bees in a social context, luckily there 

### 20 are bioassays that are routinely used in apicultural and honey 

### 21 bee biology research that can do this in vitro very high 

### 22 throughput and very readily, very reliable. For example, Evans 

### 23 et al. 2009 published a paper that just articulates the use of 

### 24 what they call bee cups, which are placing dozens, usually 

### 25 around 50 or so newly emerged adult honey bee workers into 

### 26 plastic cups, clear plastic cups where they are fed at lib with

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1 food and can be subject to various treatments, and then in an 

### 2 incubator followed over a period of time. 

### 3 So, it seems logical to the Panel who weighed 

### 4 in on this that there can be two bioassays whereby that type of 

### 5 system could capture these types of phenomenon. One is - not to 

### 6 use acute versus chronic here, but rather kind of a one-time 

### 7 application of a compound in question and the following a marked 

### 8 cohort of individuals within that social group, within that very 

### 9 small social unit over time, and then looking at their 

### 10 longevity. So, measuring longevity curves within that group of 

### 11 a one-time application of a particular compound. 

### 12 Another analogous means of doing that would be 

### 13 to feed the entire group consistently with different compounds, 

### 14 so that the entire group is constantly exposed to that and the 

### 15 following the same longevity over time to see how long the focal 

### 16 workers live. We can provide additional detail about the 

### 17 logistics of these bioassays, but I think the endpoint here is 

### 18 that studying them in a social context when it deals with honey 

### 19 bees I think is very important compared to isolating them and 

### 20 testing them in a vacuum, which has a lot less biological 

### 21 meaning to honey bees as a social system than doing it in a 

### 22 social context. 

### 23 One last point to point out again to deal with 

### 24 his particular question about just looking at the currently 

### 25 available toxicity test with looking at mortality and survival 

### 26 as endpoints is also another temporal component and that is the

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1 seasonality of honey bees themselves and how honey bees can be 

### 2 physiologically different over different times of the year. 

### 3 I'll let Dr. Ostiguy comment on this further if she wants, but 

### 4 bees in the summer are very physiologically different from bees 

### 5 in the winter. So, incorporating the types of bees that would 

### 6 actually be used in these bioassays is biologically meaningful 

### 7 and nontrivial and out to taken into consideration on this. 

### 8 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 9 **DR. MAY BERENBAUM:**Fine. The only other 

### 10 thing - no I just see Ostiguy before Berenbaum. The only other 

### 11 - I concur, would just add to that there be useful to be at 

### 12 least cognisant of the genetic differences among populations and 

### 13 races, which is a well-known biologic attribute of bees among 

### 14 bee keepers. Dog breeds differ in the susceptibility to drugs 

### 15 or any issues with races and population. 

### 16 **DR. DANIEL SCHLENK:**My apologies. Didn't 

### 17 mean to jump the gun there. My version is different than yours 

### 18 I think. Dr. Ostiguy? 

### 19 **DR. NANCY OSTIGUY:**I concur with what David 

### 20 said. One thing about the summer/winter bees that we should 

### 21 also be taken into consideration when designing Tier I studies 

### 22 is that the particular exposures that you might see are 

### 23 different. So for instance, exposure to honey dew is going to 

### 24 be a fall phenomenon. Those will only be winter bees then. So, 

### 25 you can actually very much target where you think an exposure 

### 26 might be.

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1 **DR. DANIEL SCHLENK:**Okay. Mr. Pistorius? 

### 2 **MR. JENS PISTORIUS:**For the available bee 

### 3 toxicity tests as OECD 213 to 214, I think it is important to 

### 4 take into account behavioral abnormalities that are observed in 

### 5 the laboratory tests, especially if they occur at several doses 

### 6 that are clearly below mortality in a larger number of bees. 

### 7 **DR. DANIEL SCHLENK:**Anyone else on the Panel 

### 8 want to get in on this one? Okay. Let me go to the Agency. 

### 9 Mr. DeCant, any questions of clarification? Okay. Let's move 

### 10 onto the second section then. 

### 11 **MR. JOSEPH DECANT:**Joe DeCant, EFED. 

### 12 Question 8, section B -- please comment on additional 

### 13 measurement endpoints such as growth, which should be considered 

### 14 in future modifications of Tier I test protocols, and which are 

### 15 appropriately linked to the proposed assessment endpoints. Given 

### 16 that the queen is the reproductive unit of the colony, please 

### 17 comment on methods to evaluate effects on individual queens, 

### 18 considering practical limitations of testing with queens. 

### 19 **DR. DANIEL SCHLENK:**Dr. Tarpy? 

### 20 **DR. DAVID TARPY:**This is Dave Tarpy again, NC 

### 21 State. So this is where some of those other effects that Mr. 

### 22 Pistorius was just addressing, which are kind of the endpoints 

### 23 that we didn't articulate in part A. There are many - I think 

### 24 because of the social complexity of doing these bioassays in 

### 25 groups, it actually adds a lot of different phenomenon that can 

### 26 be measured and easily quantified and adds some robustness to

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1 these types of bioassays. One case in point including the 

### 2 additional temporal component rather than just looking at 48 

### 3 hour mortality of 72 or whatever, just having kind of binary 

### 4 data like that, by looking at these social groups, measuring the 

### 5 number of individual focal bees that have been treated or not 

### 6 treated, and as they die over time, that facilitates the ability 

### 7 to look at survivorship and analysis with survival statistics. 

### 8 So in essence, that's adding that temporal 

### 9 component to these types of bioassays. So rather than just 

### 10 taking a cross section in time, you're looking over the course 

### 11 of time and being able to make important inferences based on 

### 12 that. So, if you were to just look at 48 hours or really any 

### 13 one time point, that would be a unit dimensional means of 

### 14 assessing that. But there are multiple ways that one can arrive 

### 15 at those same kinds of mortality percentages at that time point. 

### 16 So looking at the curves and the shapes of the curves can be 

### 17 incredibly informative. That's what is captured by survival 

### 18 statistics. 

### 19 So, that's one outcome that I think would be 

### 20 facilitated with really not much additional work in these 

### 21 screening bioassays to measure the entire lifespans of 

### 22 individuals and these small in vitro groups, but then 

### 23 quantifying the slopes of mortality over time would give very, 

### 24 very informative information, we believe. Again, this has been 

### 25 done in many studies in this same capacity. So, this is fairly 

### 26 well documented in the literature.

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1 Because the question also addresses 

### 2 specifically the effects on queens, and I totally agree that 

### 3 this is incredibly important. Queens, of course are the sole 

### 4 reproductive females within colonies. They're kind o the 

### 5 carrier of all the genetic material in the colony and therefore 

### 6 a real focal point. Moreover, just on a side note, in the 

### 7 apiculture industry, there are a lot of reports about problems 

### 8 with queens, although those problems are quite nebulous. It's 

### 9 very hard to define them and to pin them down, but there seems 

### 10 to be consistent issues in the management practices when it 

### 11 comes to queens. Given their importance to the social 

### 12 super-organism of honey bee colonies, it seems to be pretty 

### 13 paramount to try and quantify that phenotype and effects on the 

### 14 queen phenotype. 

### 15 There are ways of doing that, of looking at 

### 16 quantifying queen reproductive potential, not just queen 

### 17 fecundity that is egg-laying rate, but other aspects of queens 

### 18 ability to keep the colony healthy, productive and well 

### 19 populated. I won't go over all of those different measures that 

### 20 can be done, but let's just suffice it to say that there are 

### 21 more logical measures and then reproductive measures that could 

### 22 be done looking at things like the stored sperm count of the 

### 23 queens, the viability of sperm, ovarial number, egg-laying 

### 24 rates, all of those types of things. 

### 25 How those many different characters translate 

### 26 to the ultimate colony phenotype however, it not entirely known

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1 and may be difficult to make extrapolations on measuring those 

### 2 characters in an individual queen to the ultimate colony 

### 3 phenotype. So, in that sense, at a Tier I level, it may be very 

### 4 difficult to be able to quantify effects of particular compounds 

### 5 on queens and reproductives, the drone as well. So, it might 

### 6 really necessitate Tier II levels in order to be able to truly 

### 7 elucidate effects on queens and drones. That's something that 

### 8 we will get to in later questions as well. 

### 9 Some other potential measurements and 

### 10 endpoints that the discussants brought up on these in vitro 

### 11 bioassays is quantifying immunocompetence, effects of compounds 

### 12 on abilities of individuals to withstand disease or other 

### 13 challenges. While this is being studies quite heavily in the 

### 14 literature, it was pretty well agreed that we still don't have a 

### 15 very good cause effect understanding of a lot of these 

### 16 measurements such as phenoloxidase activity, encapsulation 

### 17 response, expression of antimicrobial peptides, those types of 

### 18 things. They can be done, but we don't understand the full 

### 19 relationship to the end phenotype. So, while it might be 

### 20 interesting to keep those in mind, it be premature to include 

### 21 them in a high throughput endpoint at this point. 

### 22 As was mentioned earlier just a few moments 

### 23 ago, other endpoints that could and probably should be 

### 24 considered are changes in behavior, which can be again, very 

### 25 difficult to quantify and the present their own challenges. But 

### 26 in a social organism such as honey bees, they are paramount.

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1 They are everything. So, looking at the super-organism cannot, 

### 2 almost by definition, ignore effects on behavior. 

### 3 So, measuring simple behavioral bioassays such 

### 4 at motility, social interactions, triple axis, other behavioral 

### 5 changes can be and have been, in the literature, documented. We 

### 6 also feel that in order to assist this type of data collection 

### 7 in a repeatable high throughput measure that there is a real 

### 8 need for technologies that could capture and automate that type 

### 9 of data collection. And, we know of several initiatives that 

### 10 are trying to take video recordings of social insect colonies 

### 11 and trying to automate the complexities of a lot of the social 

### 12 insect interactions that are going on. So, there ought to be a 

### 13 real premium on developing those types of technologies in order 

### 14 to incorporate them into here, but again, at this point in time, 

### 15 it might be premature. 

### 16 One behavioral bioassay that is well 

### 17 articulated in the study for decades now is and also discussed 

### 18 in the White Paper, so I won't go too much into it, but that's 

### 19 the PER and learning conditioning bioassays. This is proboscis 

### 20 extension reflex, which incorporates the natural instinct of 

### 21 honey bee worker to stick out her tongue in response to her 

### 22 antenna being touched by a small droplet of sucrose solution. 

### 23 That reflex can then be conditioned and pared with the stimulus 

### 24 such as an odor or tactile response so that over a few trials of 

### 25 that association tested individuals will then immediately stick 

### 26 out their tongues if they are exposed to that same odor. That

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1 is a means by which researchers can quantify learning in their 

### 2 ability to make this association. 

### 3 There is a very rich literature on using PER 

### 4 for learning and conditioning. These are also potential 

### 5 endpoints that can be incorporated and harnessed as part of a 

### 6 Tier I assessment. 

### 7 **DR. DANIEL SCHLENK:**Okay. Dr. Berenbaum? 

### 8 **DR. MAY BERENBAUM:**Just two additional 

### 9 comments. One is drones represent a significant data gap. 

### 10 They're sort of literally the forgotten men here. Even though, 

### 11 they may seem nonessential most of the time, they do serve as a 

### 12 vital and irreplaceable function in colony life. There's not 

### 13 much known about their toxicology. 

### 14 The other comment is that just to point out 

### 15 that the proboscis extension reflex represents a particular type 

### 16 of learning, repetitive learning, and that's only a component of 

### 17 the incredibly rich behavioral repertoire of honey bees. So, 

### 18 it's a proxy maybe. So we have a lot of links to make. First 

### 19 per has to be linked to in general to colony phenotype and in 

### 20 addition, its place within the role of very complex behavioral 

### 21 repertoire of honey bees and needs to be considered. 

### 22 **DR. DANIEL SCHLENK:**Thank you. Dr. Ostiguy? 

### 23 24 **DR. NANCY OSTIGUY:**I concur with Dr. Tarpy 

### 25 and Dr. Berenbaum. 

### 26 **DR. DANIEL SCHLENK:**Okay. Dr. Fefferman?

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1 **DR. NINA FEFFERMAN:**Hi, sorry, I feel like 

### 2 I'm talking a lot. The one thing about queens that I don't see 

### 3 discussed ant it could be completely irrelevant and this is a 

### 4 toxicology question - is either her pheromone production 

### 5 directly or the interpretation of pheromones by workers with 

### 6 higher toxicity at a sublethal effect? There's been a lot of 

### 7 debate in the literature about who is controlling what happens 

### 8 because of queen pheromone, but it doesn't really matter if that 

### 9 gets disrupted. Then that's a huge disruption in colony health 

### 10 and colony reproduction. So, I have no idea if there're 

### 11 appropriate assays for this, but my learned colleagues certainly 

### 12 know if this is an appropriate question to add and if so, how to 

### 13 measure it if it can be measured. 

### 14 **DR. DANIEL SCHLENK:**Yes, Dr. Pettis? 

### 15 **DR. JEFF PETTIS:**There is some recent 

### 16 literature - recent papers in the literature about the effect of 

### 17 disease on pheromone production, and probably you might expect 

### 18 the same thing with some pesticide exposure. Bioassays might 

### 19 include things like new behavior, the number of bees attending 

### 20 the queen, things like that, so there are bioassays that could 

### 21 be looked at. 

### 22 If I may, I'd just like to comment a bit on 

### 23 what Dr. Tarpy said about how to look at queen - or how to 

### 24 incorporate queens into Tier I. I think it's difficult because 

### 25 of the reports that we are getting of queen failures. I think 

### 26 it behooves us to try to look at ways to incorporate it. As I

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1 pointed out earlier, the queen, we believe and we don't have 

### 2 good numbers on it, but in effect, they process much more food 

### 3 than any other worker or drone in the colony because of their 

### 4 high egg laying rate. So, there could be some biomagnification 

### 5 of whatever residue is present in the colony. 

### 6 I can think of a couple of ways. With your 

### 7 group dynamics, it would be possible to just simply introduce 

### 8 queens into that environment. I don't know if it's practical to 

### 9 watch behavior, but certainly you could introduce queens and 

### 10 then see compared to control queens if there was any direct 

### 11 effect in a Tier I test. So, just adding queens the workers 

### 12 that are in those Tier Is, it's a possibility. 

### 13 Dave pointed out some of the things we look at 

### 14 in queen health, such as sperm viability. The queen stores the 

### 15 sperm, so we can actually measure after a certain period of time 

### 16 of exposure of the viability of sperm within the queen. I 

### 17 believe there are issues with queens become drone layers. So, 

### 18 they either run out of sperm or the sperm is dying. That seems 

### 19 to be a common theme of late. So, we have bioassays for that as 

### 20 far as ways of looking at queens. 

### 21 Lastly without sacrificing the queen, you can 

### 22 actually take a fecal sample from the queen and that might be 

### 23 telling. You take and analyze the fecal material. But again, 

### 24 usually at the end of a bioassay, you just sacrifice the queen 

### 25 and look at a number of parameters. I will say that while we're 

### 26 doing a lot of this, the methods and the standardization, I

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1 don't this is there at this point. 

### 2 **DR. DANIEL SCHLENK:**Yes, Dr. Hunt? 

### 3 **DR. GREG HUNT:**Hi. I'm just willing to bring 

### 4 up another potential behavioral endpoint, which is homing 

### 5 ability. There have been a number of recent studies looking at 

### 6 pesticide effects on bees' ability to find their way home. You 

### 7 can train them to a feeding station and then take them a 

### 8 distance to the side to make it a little bit more challenging to 

### 9 see who gets home. And, this has some - it's very important for 

### 10 the bees to be able to find their way home, obviously. 

### 11 Otherwise, they're toast. And it has some advantages over the 

### 12 PER assay because with the learning assays, it depends on the 

### 13 bees' sort of motivation, their inherent response, threshold to 

### 14 sucrose and things of that sort. 

### 15 **DR. DANIEL SCHLENK:**Yes, Dr. Tarpy? 

### 16 **DR. DAVID TARPY:**This is Dave Tarpy again. 

### 17 So, Greg, I totally agree with that of course. It makes a lot 

### 18 of sense at Tier II and Tier III, but these being questions 

### 19 about Tier I, would you think that there would be a good proxy 

### 20 at the Tier I level that would be able to kind of quantify 

### 21 homing behavior, kind of, you know, in an in vitro setting? 

### 22 **DR. GREG HUNT:**No, I guess I was getting 

### 23 ahead of the game there, but I'm glad you pointed that out. 

### 24 **DR. DAVID TARPY:**Well, then bring it up again 

### 25 with the Tier II. I think that will be - because that is 

### 26 obviously important.

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1 **DR. DANIEL SCHLENK:**Yes, Dr. McManaman? 

### 2 **DR. JAMES MCMANAMAN:**So, this is going to 

### 3 come up in the next session, but I just want to ask the panel. 

### 4 There's a lot of evidence that there is pretty definitive 

### 5 genetic changes that occur in queens when they become queens. 

### 6 They occur in the brain. So, I'm just wondering if genetic 

### 7 markers could be used as a way as an alternative method of 

### 8 evaluating queen health and the effects of pesticide on her 

### 9 reproductive ability. 

### 10 **DR. DAVID TARPY:**This is Dave Tarpy again. 

### 11 Actually, there's no genetic - genome of workers and queens are 

### 12 exactly the same. It's just what genes are getting turned on 

### 13 and off. That's actually a function. There have been genomic 

### 14 studies that have looked at caste determination and everything 

### 15 like that, but unless other panel members would like to correct 

### 16 me on this, I don't know if any of them would be good genetic 

### 17 markers that would be reliable to really quantify in a 

### 18 continuous way of some sort of measure for queen quality and 

### 19 queen reproductive potential, but I could be wrong on that. 

### 20 **DR. DANIEL SCHLENK:**Dr. James? 

### 21 **DR. ROSALIND JAMES:**Two comments. One in 

### 22 response to Jim. I agree with Dr. Tarpy. We're just not there 

### 23 yet, I think, in understanding. We're starting to do work with 

### 24 transcript looking at gene expression, but expression levels of 

### 25 transcriptome isn't necessarily related to what is actually 

### 26 happening physiologically and it's complex. We're just

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1 beginning to look at that. I don't think it's anywhere near 

### 2 being ready as some sort of a screening tool. 

### 3 And I had another comment. I was waiting for 

### 4 some of the queen discussion to die down because I want to 

### 5 change the subject. You can go back to queens if you want now 

### 6 that I have the mic. You asked for other endpoint measurements 

### 7 and we'll probably talk about this a little bit more later when 

### 8 we get in - you sort of re-asked the same questions in different 

### 9 ways. So we'll get to this more later, but in part A, we talked 

### 10 a little bit about using longevity, using these bioassays that 

### 11 are run for a longer time and one advantage to doing that. And 

### 12 I'm speaking now about larval bioassays instead of adults as you 

### 13 can look at development time. 

### 14 So, I think later we'll talk a little bit more 

### 15 about alternative bioassays, but one of the things you can do 

### 16 are these larval bioassays and then run them out to some 

### 17 biological endpoint such as cell capping or pupation, and then 

### 18 measure how long it takes to get to that stage for each 

### 19 individual. This would give you another endpoint measurement in 

### 20 addition to mortality. 

### 21 **DR. DANIEL SCHLENK:**Okay. Any other 

### 22 comments? I didn't hear growth mentioned at all. Is that an 

### 23 endpoint? Because that's mentioned in the question. Mr. 

### 24 Pistorius? 

### 25 **MR. JENS PISTORIUS:**One comment that is also 

### 26 related to charge question 9, I apologize if I may be a bit

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1 early in this. I think growth could be included in the future 

### 2 coming Aupinel test method or OECD guidelines, which is 

### 3 developed and regarded in the Tier I, although it needs to be in 

### 4 short, additional handling of the larvae does not affect 

### 5 mortality, which is a bit critical in this test. I think this 

### 6 is the only Tier I test method which has been or is under 

### 7 validation that would actually cover growth on a Tier I. 

### 8 **DR. DANIEL SCHLENK:**Dr. James? 

### 9 **DR. ROSALIND JAMES:**Hopefully this isn't just 

### 10 reiterating the same thing, but the larvae are extremely 

### 11 delicate and you can't really pick them out and weigh them 

### 12 regularly. At the end, perhaps you could weight them, but 

### 13 growth is difficult to measure for the larvae. And then the 

### 14 adults, you are not going to have growth occurring. So, this is 

### 15 one reason way we use development time rather than growth. 

### 16 **DR. DANIEL SCHLENK:**Yes, Dr. Fefferman? 

### 17 **DR. NINA FEFFERMAN:**Although it is - well, 

### 18 I'm sure everyone meant individual growth, the rate of change of 

### 19 the size of the colony after mating or once the queen starts egg 

### 20 laying, not just a measurement of fecundity but a successful 

### 21 rate of change - that ramp up, the differential there might 

### 22 actually be a meaningful colony health measure. 

### 23 **DR. DANIEL SCHLENK:**Yes, Dr. Ostiguy? 

### 24 **DR. NANCY OSTIGUY:**One difficulty with that 

### 25 is there are tremendous strain differences. So, if you're going 

### 26 to ask whether or not or how fast a colony builds, you'd better

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1 standardize for which type of honey bee you're using. 

### 2 **DR. DANIEL SCHLENK:**Mr. Pistorius? 

### 3 **MR. JENS PISTORIUS:**Especially regarding 

### 4 growth, we would like to discuss this also in the Tier II and 

### 5 Tier III tests because they are very reliable and very 

### 6 well-suited methods available for measuring growth. 

### 7 **DR. DANIEL SCHLENK:**Okay. Any other comments 

### 8 on this section? Dr. Delclos? 

### 9 **DR. KENNETH DELCLOS:**Question - now I think 

### 10 that in the first part A, I think we are talking about using a 

### 11 single dose. How critical is the timing of the dose for these 

### 12 various - I mean, I don't know anything about the development of 

### 13 metabolic capacity. I don't hear much about that toxicokinetics 

### 14 discussed in here, but also in the response if you're looking at 

### 15 a specific response, how tight a window - is there an apriority 

### 16 way of choosing one that should be administered? 

### 17 **DR. DAVID TARPY:**This is David Tarpy. I 

### 18 don't think there is an apriority way, but often times in these 

### 19 bioassays, they're done by newly closed workers, newly emerged 

### 20 workers as they're placed into these experimental arenas, but 

### 21 it's true. There may be differences in the time of application 

### 22 on their effects. 

### 23 **DR. DANIEL SCHLENK:**Okay. Any other comments 

### 24 for question 8? Okay. I'm going to go to the Agency - 

### 25 questions of clarification? Mr. DeCant? 

### 26 **MR. JOSEPH DECANT:**Yeah, this is Joe DeCant

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1 from EFED. I just have a question. It sounds like - thank you 

### 2 to the Panel. We got some good feedback in terms of the number 

### 3 of different endpoints. I think some of the endpoints like Dr. 

### 4 Tarpy, you mentioned about immunocompetence. It sounds like 

### 5 some of the endpoints are quite there and aren't quite there and 

### 6 aren't quite developed yet to be able to include into some of 

### 7 our studies. I also want to ask a clarification question - some 

### 8 of the endpoints that were mentioned like the PER Assay - I know 

### 9 that we are trying to link these endpoints with assessment 

### 10 endpoints. So survival, mortality and to what extent do you 

### 11 think some of these measurement endpoints are actually linked to 

### 12 our assessment endpoints? 

### 13 **DR. DANIEL SCHLENK:**Dr. Tarpy? 

### 14 **DR. DAVID TARPY:**This is Dave Tarpy again. I 

### 15 would argue all of them to greater or lesser degrees. Again, 

### 16 adding that extra layer of biological complexity, that is the 

### 17 colony, inherently brings in all of these different aspects. I 

### 18 think logistically in measuring them in a repeatable, 

### 19 quantifiable way is really a difficulty. And as you're correct 

### 20 at that immunocompetence measures may not be as well elucidated 

### 21 to be able to make those connections to the social level yet, 

### 22 but it is definitely something to keep in mind. Some of the 

### 23 other bioassays like the PER are more robust and standardized. 

### 24 So, we bring all of them up as a means of 

### 25 keeping them in mind as to complexity of dealing with the social 

### 26 organism rather than a solitary one, but that there is an

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1 understanding that just based on feasibility that not all of 

### 2 them are going to be incorporated or at least right away. 

### 3 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 4 **DR. MAY BERENBAUM:**With that said, I think 

### 5 it's important to - at least one component of your question is 

### 6 the linkage to colony phenotype. And, I don't even think for 

### 7 proboscis extension reflex that there is a bulletproof linkage. 

### 8 So, there is another data gap. 

### 9 **DR. DANIEL SCHLENK:**Yes, Mr. Pistorius? 

### 10 **MR. JENS PISTORIUS:**I still think it is very 

### 11 difficult to actually link the findings of the PER Test to the 

### 12 realistic situation that bee colonies are encountered with. I 

### 13 would like to point out that there is an area of major need for 

### 14 research to make this link to actually establish a link and only 

### 15 very limited number of studies that have looked into it, but 

### 16 with those for instance, we could find some effect in PER, we 

### 17 could not find any relation as to foraging behavior or to honey 

### 18 flow or honey yield of the colonies. But again, there is so 

### 19 much limited data available on how to link those two that I 

### 20 think we're a little bit unsure about this. 

### 21 **DR. DANIEL SCHLENK:**Dr. Tarpy? 

### 22 **DR. DAVID TARPY:**This is Dave Tarpy again. I 

### 23 failed to mention last time, and I agree with all of the points 

### 24 that were raised there is because of these difficulties and 

### 25 linkages to full colony phenotypes, it places a real premium and 

### 26 a priority on Tier II and III bioassays, and that it is almost

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1 inherent to study a super-organism at the level of that 

### 2 super-organism, which is the colony. So, it's incredibly 

### 3 difficult to use Tier I in vitro tests to truly adequately 

### 4 capture the entire biology at the colony level. So that's an 

### 5 inherent difficulty seen here, which emphasized Tier II and Tier 

### 6 III bioassays for these screenings. 

### 7 **DR. DANIEL SCHLENK:**Okay. Dr. Ostiguy? 

### 8 **DR. NANCY OSTIGUY:**I'm going to disagree a 

### 9 little bit with some of the comments. I actually think the PER 

### 10 does provide some very useful information that can be related 

### 11 then to the full colony in the same way that we evaluate muscle 

### 12 rechemical exposure, et cetera, on human learning ability that 

### 13 we then apply to a population. So, if you adversely affect 

### 14 individuals within the group, which is what we're looking at 

### 15 with honey bees, you are going to at some point impact the whole 

### 16 group. So to me, there's the connection, you know. The 

### 17 individuals do make up the whole. And if we interfere with the 

### 18 ability of individual bees to learn, then we interfere with the 

### 19 whole. So, what we might need to do with the PER is set a 

### 20 particular standard for the degree of learning inhibition rather 

### 21 than saying we can't use the PER as a Tier I screen to try to 

### 22 get at some of that some of that social insect interaction. 

### 23 **DR. DANIEL SCHLENK:**Okay. Dr. Fefferman? 

### 24 **DR. NINA FEFFERMAN:**We'll talk more about 

### 25 this at the end of everything, but I think that kind of question 

### 26 is exactly the sort of thing where modeling can help.

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1 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 2 **DR. JEFF PETTIS:**Just in regard to the PER 

### 3 and other tests that we might run, I agree that the PER has been 

### 4 vetted well as far as demonstrating effects, but I think I would 

### 5 have more confidence in something like Dr. Hunt suggested in 

### 6 these returning forager assays, but it may not be exactly a Tier 

### 7 I. It may be closer to a Tier II test where you actually dose 

### 8 foragers at realistic exposure levels and then ask them to 

### 9 return to the hive. I think there have been several papers on 

### 10 that of late. They seem very convincing that realistic field 

### 11 exposure can disrupt returning forager behavior. You can see 

### 12 that that can have direct effects on the colony. 

### 13 **DR. DANIEL SCHLENK:**Okay. Dr. Klaine? Dr. 

### 14 Ostiguy? 

### 15 **DR. NANCY OSTIGUY:**I actually agree, but I 

### 16 think Tier I -- using PER as a Tier I would then trigger us to 

### 17 do that field returning ability. So, the PER could be set 

### 18 sufficiently conservative so that if we need to, we can then 

### 19 check to see if it really has that effect on the whole group. 

### 20 **DR. STEPHEN KLAINE:**This is Steve Klaine with 

### 21 Clemson. Thinking about this on a larger scale in terms of 

### 22 trying to move from a single organism to a more complex 

### 23 super-organism, it seems to me that any one behavior may not get 

### 24 us all the way, but that a bank of these behaviors may give us a 

### 25 lot of insight into the whole hive health by using single 

### 26 organisms. So, I think it ultimately - and I agree with the

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1 discussions last night - we're not there yet and what you said 

### 2 today, but I think that's where we need to be if we're going to 

### 3 start making predictions on hive health. 

### 4 **DR. DANIEL SCHLENK:**Anyone else to weigh in 

### 5 on this one? No? I hear some discussions back there. Do you 

### 6 guys need some more clarification or are we. 

### 7 **MR. JOSEPH DECANT:**This is Joe DeCant from 

### 8 EFED. Could you elaborate a little bit on the bank of behaviors 

### 9 that you mentioned about and what type of behaviors will be 

### 10 helpful? I know Dr. Pistorius, you mentioned about some of the 

### 11 behavioral effects that are observed in the OECD Guidelines, and 

### 12 so if you could clarify or elucidate on those behaviors. 

### 13 **MR. JENS PISTORIUS:**Well there are a number 

### 14 of described categories of behavioral abnormalities, and it is 

### 15 quite difficult to now put really into a frame when this would 

### 16 actually trigger a real additional concern. But for instance, 

### 17 during my work, I was encountered with one substance that had 

### 18 doses clearly below the lethal doses at a number of dosings, 

### 19 also in very low dosings, in different laboratories, repeatable 

### 20 effects, severe effects on behavior of very agitated, very 

### 21 nervous, trembling, shaking, all this. So, I would just like to 

### 22 point out is that it is important that the assessed behavioral 

### 23 abnormalities are reported by the contractors and that the risk 

### 24 assessor also regardless of any trigger values still has the 

### 25 flexibility to say, on the basis of these concerns, we have to 

### 26 go to a higher tier testing and do additional testing. Does

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1 that answer your question? 

### 2 **DR. DANIEL SCHLENK:**Any other questions, Mr. 

### 3 DeCant? Oh, I'm sorry, Dr. Klaine has something to add here. 

### 4 **DR. STEPHEN KLAINE:**Yeah, I was just - you 

### 5 talked about the bank - I mentioned a bank of behaviors. I 

### 6 think there are a lot of them that maybe aren't standardized but 

### 7 at least in my discussions with my bee colleagues, have really 

### 8 been intriguing. The discussion we had last night on the 

### 9 nursing, the in vitro nursing assay, where you could - that's 

### 10 another behavior that you can quantify, because in my mind, at 

### 11 least, it's not one behavior that describes the health of the 

### 12 hive, but its culmination of all these behaviors that make these 

### 13 hives work. So, if you can capture, quantify three or four of 

### 14 them, you might get a lot better insight into bee health or hive 

### 15 health. 

### 16 **DR. DANIEL SCHLENK:**Okay. Anyone else? Let 

### 17 me go back to Mr. DeCant. Do you have any other questions of 

### 18 clarification. 

### 19 **DR. JOSEPH DECANT:**No, I think we're good. 

### 20 **DR. DANIEL SCHLENK:**Great. Okay. Let's go 

### 21 ahead and move onto question 9. Want to read letter A into the 

### 22 record for us? 

### 23 **MR. JOSEPH DECANT:**Joe DeCant, EFED. 

### 24 Question 9, subpart (a) - please comment on the extent to which 

### 25 the Aupinel et al. in vitro method serves as an appropriately 

### 26 conservative estimate of Tier I acute oral exposure of honey bee

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1 larvae to pesticides, given differences in this test design from 

### 2 actual in-hive exposure conditions, for example during the first 

### 3 three days of the larval development stage, larvae consume royal 

### 4 jelly and brood food, and the uncertainty regarding the extent 

### 5 to which larvae rely exclusively on pollen or nectar as opposed 

### 6 to royal jelly or brood food. 

### 7 **DR. DANIEL SCHLENK:**Our lead discussant for 

### 8 this question is Dr. Berenbaum. 

### 9 **DR. MAY BERENBAUM:**Determining toxicity of 

### 10 pesticides to larvae using in vitro testing is complicated by 

### 11 several factors including possible genetic differences among 

### 12 colonies and difficulties synchronizing larval age, both of 

### 13 which were mentioned by Aupinel and also included in the White 

### 14 Paper. In addition as yet uncharacterized attributes of royal 

### 15 jelly, brood food, honey and been bread may be relevant to 

### 16 pesticide tolerance in larvae. 

### 17 Royal jelly, for example, contains substances 

### 18 that inhibit DNA methylation and histone deacetylase activity, 

### 19 also promote gene expression and similarly honey and bee bread 

### 20 constituents contain, including phytochemicals that can regulate 

### 21 detoxification pathways in worker bees at least. These factors 

### 22 have the potential to alter responses to pesticides, but without 

### 23 more complete understanding of how the composition of food 

### 24 affects toxicity or bioavailability despite compound, it's 

### 25 difficult to say conclusively that the method is conservative, 

### 26 although available information points in that direction.

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1 One specific concern is that the proposed 

### 2 method for testing larvae is based on pesticide levels in pollen 

### 3 and nectar administered to day-5 larvae. Day-5 larvae destined 

### 4 to be queens eat royal jelly. If royal jelly has 1/100th the 

### 5 level of pesticides contaminants the nectar and pollen, test 

### 6 conducted in Tier I using maximum concentrations of pesticides 

### 7 based on worker exposure will be much higher than those that are 

### 8 experienced by queen larvae. 

### 9 So, in terms of toxicity, this approach is 

### 10 reasonable, but if caste determination and hive viability are 

### 11 concerns, then these pesticide amounts might not be appropriate. 

### 12 As noted earlier too, the assumption that royal jelly 

### 13 concentrations of pesticides are 1/100th those in other bee 

### 14 foods, those estimates are based on one published and one 

### 15 unpublished study. So, that underlines that assumption. 

### 16 So, the impact of larval diet on pesticide 

### 17 detoxification represents a significant data gap. I forgot to 

### 18 introduce my comments by saying I am extremely grateful that the 

### 19 EPA is undertaking this effort and also to point out that the 

### 20 first indication that pesticides used for agricultural purposes 

### 21 have nontarget impacts on bees goes back to 1895, and the fact 

### 22 that we are now a century later talking about major data gaps, I 

### 23 think is unsettling at the least, and that's one reason I'm very 

### 24 grateful to the EPA for convening this Panel. 

### 25 Okay. So, data gap - the artificial diet used 

### 26 for bioassay contains glucose and fructose as proxies for honey,

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1 but honey is substantially more than just carbohydrates. 

### 2 Phytochemicals of honey may well influence pesticide toxicity 

### 3 and metabolism as they do in other herbivorous insects. There 

### 4 is a rich literature in the effect of phytochemicals and 

### 5 detoxification in other herbivores. But in larval bees, it's 

### 6 essentially nonexistent. 

### 7 More generally, the concept of an acute dose 

### 8 for a larva may be biologically difficult to interpret. Unlike 

### 9 workers who may experience one-time encounters through pesticide 

### 10 drift outside the hive, larvae mature continuously provide food 

### 11 that has been already processed by workers seem less likely to 

### 12 have acute exposures more over because larvae are confined 

### 13 during their development, they're subject to continuous contact 

### 14 with wax, which if contaminated is likely to be a source of 

### 15 toxicity that is not directly assessed by the proposed methods. 

### 16 17 Contact bioassays would be very useful in this 

### 18 context, particularly using pupae just for operational and 

### 19 logistic reasons and also there are precedents for pupal 

### 20 bioassays of microbial toxicity and the like. I guess those are 

### 21 my comments. 

### 22 **DR. DANIEL SCHLENK:**Okay. Thanks. Dr. 

### 23 McManaman? 

### 24 **DR. JAMES MCMANAMAN:**My comments were 

### 25 incorporated into Dr. Berenbaum's. 

### 26 **DR. DANIEL SCHLENK:**Dr. Ostiguy as well?

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1 **DR. NANCY OSTIGUY:**I concur, my comments were 

### 2 included by Dr. Berenbaum. 

### 3 **DR. DANIEL SCHLENK:**Okay. Open to other 

### 4 Panel members to weigh in on this one. Yep, Dr. James? 

### 5 **DR. ROSALIND JAMES:**I would have to weigh in 

### 6 on the larval bioassays. Because I do so many of them myself, 

### 7 although not with honey bees, but in line with what Dr. 

### 8 Berenbaum said about the one time acute exposure, one of the 

### 9 things that we talk about is contaminating all of the food that 

### 10 you feed to the larvae. So, during the full five-day feeding 

### 11 period, the bees would be continually fed with pesticide 

### 12 contaminated food stuff instead of a one-time dose. This would 

### 13 also get at the question about which age is most sensitive in 

### 14 the developing larvae, which size increase is quite amazing, and 

### 15 we don't really know about sensitivity of the different ages. 

### 16 But I think it would be more realistic to have - they would 

### 17 continually be fed pesticides. 

### 18 **DR. DANIEL SCHLENK:**Any other comments? Just 

### 19 curious on my own side here is would you propose then a 

### 20 standardized diet that would have the same reference material in 

### 21 terms of phytochemical input that could be a baseline 

### 22 standardized diet that would used? Is that something that you 

### 23 would propose? 

### 24 **DR. MAY BERENBAUM:**This is May Berenbaum. It 

### 25 would be ideal. It would be preferable. What constituents 

### 26 would go into that diet would be probably subject to

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1 considerable discussion. 

### 2 **DR. DANIEL SCHLENK:**Okay. Anything else on 

### 3 A? Oh, sorry. 

### 4 **DR. JAMES MCMANAMAN:**I have a question for 

### 5 the Panel. This is just out of ignorance - is royal jelly royal 

### 6 jelly? I mean, are the constituents of royal jelly independent 

### 7 of strain of kind of bee? 

### 8 **DR. ROSALIND JAMES:**This is Rosalind James. 

### 9 I'm going to let one of the honey bee folks answer that 

### 10 question, but I have to add that you do want to make sure that 

### 11 when you buy the royal jelly it's not all ready contaminated 

### 12 with pesticides. 

### 13 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 14 **DR. JEFF PETTIS:**Jeff Pettis. I was going to 

### 15 say a solitary person commenting on royal jelly, come on Rose. 

### 16 That was going to be my point was that royal jelly is royal 

### 17 jelly by and large, but you really have to consider the source 

### 18 and we've had major issues in doing lab studies and even trying 

### 19 to do any kind of larval rearing in that we get contamination in 

### 20 the royal jelly, so that the batch of royal jelly has to be 

### 21 tested and be standardized and known to be clean before it's 

### 22 used. Otherwise, you get confounding results. 

### 23 **DR. DANIEL SCHLENK:**Dr. Fefferman and Dr. 

### 24 Hunt? 

### 25 **DR. GREG HUNT:**I was just about to say that 

### 26 that was an excellent question. I don't really know what the

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1 variability is in royal jelly, but ideally, we would have an 

### 2 artificial diet. It seems that just doesn't work very well. 

### 3 **DR. DANIEL SCHLENK:**That was Dr. Hunt. Dr. 

### 4 McManaman? Hold on, let me get Dr. McManaman back. 

### 5 **DR. JAMES MCMANAMAN:**Yeah, so my question was 

### 6 - you bring up some important points about contamination, but my 

### 7 question was related to the charge question is that if we're 

### 8 trying to understand the effects of pesticides and the food can 

### 9 interfere or affect their responses and royal jelly is a 

### 10 component of that, especially for queens in early development, 

### 11 then I think it's important to know whether it really is the 

### 12 same thing because there are factors in royal jelly that can 

### 13 influence strain differences. 

### 14 **DR. DANIEL SCHLENK:**Okay. Dr. Fefferman? 

### 15 **DR. NINA FEFFERMAN:**Sorry, thanks. I still 

### 16 have the same question I had before, which is when you say 

### 17 contamination in the royal jelly that you're buying, is it 

### 18 pesticide contamination? Because from listening to the 

### 19 conversation just a bit ago, I was under the impression that 

### 20 royal jelly pesticide contamination was really sort of the 

### 21 lowest level of concern in terms of what they were eating, and 

### 22 that sounds scary. 

### 23 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 24 **DR. JEFF PETTIS:**Probably one of the major 

### 25 contaminants is antibiotics. You can get antibiotic 

### 26 contamination. Beekeepers use antibiotics in the colony to

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1 control bacterial diseases. I think 80 percent or 90 percent of 

### 2 the world's production of royal jelly comes from China, and so 

### 3 you can have issues there. You need to really get your royal 

### 4 jelly from colonies that are managed almost totally organic. 

### 5 You also can get some miticide compounds that are used by 

### 6 beekeepers to control mites that get incorporated in the royal 

### 7 jelly. 

### 8 **DR. DANIEL SCHLENK:**Dr. Ostiguy and then Dr. 

### 9 Hunt? 

### 10 **DR. NANCY OSTIGUY:**We also need to be 

### 11 concerned about the possible pathogens that are in that royal 

### 12 jelly. 

### 13 **DR. DANIEL SCHLENK:**Dr. Hunt? 

### 14 **DR. GREG HUNT:**I was just going to comment 

### 15 that it might be best to collect your royal jelly fresh and then 

### 16 just make sure that the controls and the treated get the same 

### 17 stuff. 

### 18 **DR. DANIEL SCHLENK:**Okay. Any other comments 

### 19 on 9A? Mr. DeCant, do you have all you need for 9A there? 

### 20 **MR. JOSEPH DECANT:**I do. 

### 21 **DR. DANIEL SCHLENK:**Okay. All right, let's 

### 22 move on to the second question of 9. 

### 23 **MR. JOSEPH DECANT:**Joe DeCant, EFED, question 

### 24 9, subpart (b) please comment on the extent to which pesticides 

### 25 would be more or less bioavailable using the synthetic matrix 

### 26 relied on for feeding developing bees in this in vitro method.

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1 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 2 **DR. MAY BERENBAUM:**Bioavailability will be 

### 3 influenced by the nature of the compound, larval metabolism, 

### 4 possibly food composition. Without accurate information about 

### 5 how these factors influence bioavailability within honey bee 

### 6 larvae, it's not really possible to really answer this question. 

### 7 Aupinel et al. 2007 used dimethoate, which is an 

### 8 organophosphate the highly water soluble. It's not clear how 

### 9 pesticides with different structures and properties will perform 

### 10 in this particular synthetic matrix. 

### 11 **DR. DANIEL SCHLENK:**Okay. Dr. McManaman? 

### 12 **DR. JAMES MCMANAMAN:**I have nothing to add. 

### 13 **DR. DANIEL SCHLENK:**And Dr. Ostiguy? 

### 14 **DR. NANCY OSTIGUY:**I concur with what Dr. 

### 15 Berenbaum has said all ready. 

### 16 **DR. DANIEL SCHLENK:**Anyone else on this 

### 17 section? Okay. Seems pretty clear to the point. Any 

### 18 clarification question there Mr. DeCant? 

### 19 **MR. JOSEPH DECANT:**I think we're good. 

### 20 **DR. DANIEL SCHLENK:**I'm hoping so. Okay. 

### 21 We're going to move on to C please. 

### 22 **MR. JOSEPH DECANT:**Joe DeCant, EFED, question 

### 23 9, subpart (c) - please comment on the extent to which the 

### 24 absence of trophallaxis that is the transfer of food or fluids 

### 25 between colony members, may render larvae more or less 

### 26 vulnerable to pesticides.

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1 **DR. DANIEL SCHLENK:**And Dr. Berenbaum? 

### 2 **DR. MAY BERENBAUM:**You thought B was straight 

### 3 forward. In response to the question, contribution of 

### 4 trophallaxis to pesticide resistance or susceptibility is to our 

### 5 knowledge completely and utterly unknown. This represents a 

### 6 data chasm. 

### 7 **DR. DANIEL SCHLENK:**Okay. Dr. McManaman? 

### 8 **DR. JAMES MCMANAMAN:**A very deep chasm, I 

### 9 totally concur. 

### 10 **DR. DANIEL SCHLENK:**And Dr. Ostiguy? 

### 11 **DR. NANCY OSTIGUY:**I have to commend the EPA 

### 12 for even asking the question, because we truly have no idea. 

### 13 **DR. DANIEL SCHLENK:**Yes, Dr. Berenbaum? 

### 14 **DR. MAY BERENBAUM:**Let me add that it's 

### 15 important. So, it is important to find out. 

### 16 **DR. DANIEL SCHLENK:**Yes, Dr. Tarpy? 

### 17 **DR. DAVID TARPY:**And not to beat the dead 

### 18 horse, but I don't want to move on too quickly. And that is I 

### 19 think this again underscores the complexities of all of the 

### 20 social interactions when we're dealing with a super-organism as 

### 21 opposed to just individual larvae or individual adults. 

### 22 Everything is interconnected in these types of social systems. 

### 23 So, these are important questions to ask, but it just 

### 24 underscores our data gap and the complexity of the social 

### 25 organism. 

### 26 **DR. DANIEL SCHLENK:**Dr. James, did you have

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1 something? 

### 2 **DR. ROSALIND JAMES:**Well, the only thing I 

### 3 would add is, of course, increases of spread of pesticides that 

### 4 is collected and then it may end up getting fed to many more 

### 5 individuals than originally come into contact with it. In that 

### 6 way, it would perhaps dilute the pesticide, but spread it out 

### 7 among more individuals. 

### 8 **DR. DANIEL SCHLENK:**Okay. Any other comments 

### 9 on C? Mr. DeCant, did you get what you needed on that one? 

### 10 **MR. JOSEPH DECANT:**Yeah, I think we're good. 

### 11 Thanks. 

### 12 **DR. DANIEL SCHLENK:**All right. Then read 

### 13 letter D into the record please. 

### 14 **MR. JOSEPH DECANT:**Joe DeCant, question 9, 

### 15 subpart (d) - please comment on alternative methods for testing 

### 16 individual larvae that may be appropriate for quantitative use 

### 17 in a Tier I screening-level assessment. 

### 18 **DR. DANIEL SCHLENK:**Okay. Dr. Berenbaum? 

### 19 **DR. MAY BERENBAUM:**Literature reports of 

### 20 alternative methods for testing larvae quantitatively are 

### 21 vanishingly rare. Atkins and Kellum in 1986 treated larvae in a 

### 22 colony by micropipetting pesticides directly into the cells, but 

### 23 that might quantify more as a Tier II rather than Tier I. Pupal 

### 24 bioassays have not been widely used for pesticides toxicity 

### 25 testing, but they have been used in other contexts, and it seems 

### 26 that they may be productively employed in this context,

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1 particularly given the confounding problems of social context. 

### 2 I mean, well it said people live alone and die alone - the same 

### 3 is true for bees, but they also pupate alone. So, there is one 

### 4 aspect of their life that is independent, more or less 

### 5 independent of a social context. So, that might be particularly 

### 6 suitable for Tier I. 

### 7 Let's see - following up on Dr. Tarpy's 

### 8 suggestions about other endpoints, larval transcriptomes of 

### 9 honey bees have been described. The existence of such data 

### 10 provides quantitative measures of effects of toxins on larval 

### 11 development that might be useful as alternative indicators. 

### 12 Genetic resources could allow the use of gene expression, 

### 13 detoxification, antioxidant enzyme activities, senescence traits 

### 14 as biomarkers, but in fact, the Johnson et al. 2009 suggested 

### 15 the use of ribosomal RNA fragments as a biomarker, but such 

### 16 methods would require vetting and validation before they could 

### 17 be used in Tier I screening. 

### 18 **DR. DANIEL SCHLENK:**Okay. Dr. McManaman? 

### 19 20 **DR. JAMES MCMANAMAN:**My comments were 

### 21 incorporated in Dr. Berenbaum's response. 

### 22 **DR. DANIEL SCHLENK:**And Dr. Ostiguy? 

### 23 **DR. NANCY OSTIGUY:**I concur with what's been 

### 24 said. 

### 25 **DR. DANIEL SCHLENK:**Okay. Other Panel 

### 26 members? Dr. James?

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1 **DR. ROSALIND JAMES:**I'm going to call on Dr. 

### 2 Tarpy because last night he mentioned a cage study where you put 

### 3 the brood with a few workers. We're getting to that? All 

### 4 right. 

### 5 **Dr. DANIEL SCHLENK:**I think that's for 

### 6 question 10 right? I think they're going to get to that later. 

### 7 Anyone else for D? New alternatives? Okay. Mr. DeCant, any 

### 8 questions of clarification? 

### 9 **MR. JOSEPH DECANT:**Joe DeCant, EFED. Maybe 

### 10 one question, Dr. Berenbaum, you mentioned about pupal 

### 11 bioassays. Do you have references for these protocols? Have 

### 12 they been around for a while? 

### 13 **DR. MAY BERENBAUM:**I also defer to Dr. Tarpy. 

### 14 **DR. DAVID TARPY:**This is Dave Tarpy. I don't 

### 15 have them off the top of my head. All I know is that there are 

### 16 - well, we're writing a paper right now looking at that. I know 

### 17 that many of those bioassays are also being utilized in the 

### 18 Beltsville Lab. So, we can dig those up for you for sure, yeah. 

### 19 20 **DR. DANIEL SCHLENK:**Yeah, it sounds like if 

### 21 you could provide references for some of those, it would be 

### 22 good. Dr. James? 

### 23 **DR. ROSALIND JAMES:**But this could only be a 

### 24 contact toxicity bioassay. 

### 25 **DR. DANIEL SCHLENK:**Okay. All right, let's 

### 26 go ahead and move onto letter E.

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1 **MR. JOSEPH DECANT:**Joe DeCant, EFED, question 

### 2 9, subpart (e) - typically acute toxicity tests are concluded 

### 3 between 48 and 96 hours. Please comment on the appropriate 

### 4 duration of toxicity tests for assessing acute toxicity to 

### 5 individual larval and adult bees. 

### 6 **DR. DANIEL SCHLENK:**And Dr. Berenbaum. 

### 7 **DR. MAY BERENBAUM:**The problems interpreting 

### 8 larval survival over a two to four day period is a metric of 

### 9 toxicity. The four day endpoint represents 80 percent of the 

### 10 period of active feeding. So, distinguishing between acute and 

### 11 chronic becomes difficult. Some agents induce toxicity due to 

### 12 the larval development, which would be over looked in the 

### 13 proposed approach. In general, biological transitions such as 

### 14 pupation provide a more reliable consistent and interpretable 

### 15 endpoint. Time to pupation, pupation rate and pupal weights are 

### 16 more biologically meaningful. Queens and drones can be assessed 

### 17 separately. 

### 18 Such tests however, have not yet been vetted 

### 19 thoroughly. Assessing acute toxicity over seven days for the 

### 20 present time will allow the EPA to harmonize in the national 

### 21 efforts at the OECD level until a satisfactory standard pupation 

### 22 assay has been developed, but developing such a test should be a 

### 23 priority. Among other things, these assays will be more 

### 24 sensitive to pesticide impacts on growth rate and development, 

### 25 such as IGRs. 

### 26 **DR. DANIEL SCHLENK:**Thank you. Dr.

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1 McManaman? 

### 2 **DR. JAMES MCMANAMAN:**I concur. 

### 3 **DR. DANIEL SCHLENK:**Dr. Ostiguy? 

### 4 **DR. NANCY OSTIGUY:**I concur. 

### 5 **DR. DANIEL SCHLENK:**Other Panel members? 

### 6 Nothing further to add? Wow, that's a shocker. Okay. Oh 

### 7 sorry. Kiss of death on there. Mr. Pistorius? 

### 8 **MR. JENS PISTORIUS:**Thank you. One thing to 

### 9 add regarding the in vitro test, I think it is important that 

### 10 also additional standards are testing. Now in the reference, it 

### 11 has mainly mentioned dimethoate. Also fenoxycarb would be 

### 12 necessary, so that should be principally considered in the 

### 13 guidelines. 

### 14 **DR. DANIEL SCHLENK:**Okay. Anyone else? 

### 15 Okay. Mr. DeCant, any questions of clarification? 

### 16 **MR. JOSEPH DECANT:**No more questions. 

### 17 **DR. DANIEL SCHLENK:**Oh, hold on. Ms. Garber 

### 18 has something. 

### 19 **MS. KRIS GARBER:**Kris Garber. Dr. Berenbaum, 

### 20 you indicated that it be more relevant to use the seven-day 

### 21 duration? And that's what we've been calling the chronic 

### 22 toxicity test? So does that mean that you're suggesting the we 

### 23 kind of eliminate the ideas of acute and chronic for larvae and 

### 24 just have one, kind of one risk quotient, one toxicity test? 

### 25 **DR. MAY BERENBAUM:**This is May Berenbaum. 

### 26 This is the conclusion we reached yesterday. It's just

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1 biologically hard to interpret and acute exposure of a larva. 

### 2 So, given the relative return on investment in terms of, you 

### 3 know, information that's relevant to deciding about risk and on 

### 4 whether to proceed to Tier II. The general consensus, at least 

### 5 among the discussants last night is that maybe just time to 

### 6 pupation, that interval is more biologically informative. I 

### 7 don't know if anyone else wants to weigh in here. 

### 8 **DR. DANIEL SCHLENK:**Mr. Pistorius? 

### 9 **MR. JENS PISTORIUS:**To now, the in vitro 

### 10 method is described especially for the period of egg or young 

### 11 larvae until the phase of pupation. But by doing this and 

### 12 ending there, you may get a toxicity for the larvae when they 

### 13 are white, but we know for some substances like fenoxycarb, 

### 14 which has been shown to be critical under practical conditions 

### 15 that those affect after pupation before emergence. So, by just 

### 16 looking at the first seven days, you will miss this effect and 

### 17 this has been an effect, which has been critical in practices. 

### 18 This is not developed yet in the in vitro method. It has not 

### 19 been validated. So, this is a major issue that should be pushed 

### 20 further immediately, that this test can be extended. Instead, 

### 21 there are several endpoints in this test. One would be the 

### 22 seven-day exposure and the larval mortality. Then the second 

### 23 one is the time until hatch as the endpoint, the second one. 

### 24 **DR. DANIEL SCHLENK:**Okay. Dr. Ostiguy? 

### 25 **DR. NANCY OSTIGUY:**Basically, at least my 

### 26 understanding of the conversation was that you would get the

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1 information about acute toxicity without actually having to run 

### 2 a separate test. So, you then would be able to, right now, 

### 3 what's being proposed and certainly is acceptable because it's 

### 4 been tested and is reliable, is measurements to the seven days. 

### 5 Then, as soon as we get information that allows us to take the 

### 6 larva to pupation and then to emergence, those would be good 

### 7 endpoints to strive for. So, method development is needed for 

### 8 the latter two pieces. 

### 9 **DR. DANIEL SCHLENK:**Okay. Dr. James? Yep. 

### 10 11 **DR. ROSALIND JAMES:**I'll just add, just in 

### 12 case it's not clear. If you're going to do development time, 

### 13 you'll be measuring the larvae every single day anyway. So, 

### 14 you'll still have your 48 and 96 hour mortality measures in 

### 15 there because you need to do that for your survival time 

### 16 analysis. 

### 17 **DR. DANIEL SCHLENK:**Any other questions from 

### 18 the Agency? We're good? Okay. Let's go ahead and take a break 

### 19 right now. We will convene at 20 till. 

### 20 (Brief recess.) 

### 21 Mr. DeCant, you want to read in question 

### 22 10(a)? 

### 23 **MR. JOSEPH DECANT:**Joe DeCant, EFED, question 

### 24 10, subpart (a) - please comment on the conclusion that adequate 

### 25 procedures have not been sufficiently developed and validated 

### 26 for assessing chronic toxicity to individual bees in a risk

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1 assessment context. 

### 2 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 3 **DR. MAY BERENBAUM:**The concept of the 

### 4 individual adult bee presents a challenge to assessing toxicity 

### 5 in Apis mellifera because of the importance of the social 

### 6 context, age-related polyethism over time is circumvented in the 

### 7 absence of a social medium in foragers are hard-to-keep alive 

### 8 individually. However, exposing emerging workers for 10 days at 

### 9 low doses with known ages of bees has been attempted according 

### 10 to OECD 213 with some success. 

### 11 Cage brood tests and broodless bee cup tests 

### 12 may also be employed productively in this context. So, it's 

### 13 important to emphasize that, although there may not be at the 

### 14 moment entirely reliable and interpretable assays assessing 

### 15 chronic toxicity is nonetheless remains very important. 

### 16 **DR. DANIEL SCHLENK:**Okay. Thanks. Our next 

### 17 discussants, Dr. - where am I - Dr. James, yes? 

### 18 **DR. ROSALIND JAMES:**That's me, Rosalind 

### 19 James. I concur. I would just add that we had some discussion 

### 20 about this on the previous question about possible chronic tests 

### 21 that could be done in lieu of doing actually the acute tests. 

### 22 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 23 **DR. JEFF PETTIS:**I concur with what's been 

### 24 said. 

### 25 **DR. DANIEL SCHLENK:**Dr. Ostiguy? 

### 26 **DR. NANCY OSTIGUY:**I concur.

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1 **Dr. DANIEL SCHLENK:**Other panel members want 

### 2 to weigh in on this one? Okay. Mr. DeCant, do you have what 

### 3 you need on that one? Okay. Let's move onto 10B. Want to read 

### 4 that in? I have to admit, that was Keith Sappington's 

### 5 recommendations. 

### 6 **MR. JOSEPH DECANT:**Joe DeCant, EFED -- please 

### 7 comment on the potential use of the 10 day adult worker and 

### 8 seven-day in vitro larval toxicity tests discussed in the White 

### 9 Paper for assessing chronic toxicity once these methods are 

### 10 fully vetted. 

### 11 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 12 **DR. MAY BERENBAUM:**The optimal design, we 

### 13 feel, for a larval in vitro test would be one that encompasses 

### 14 the entire active feeding period through pupation with time to 

### 15 pupation, pupal weight, percent pupation, as quantifiable and 

### 16 interpretable measurements of pesticide impact. Such a test can 

### 17 provide data on both acute and delayed responses. It goes 

### 18 without saying, but I will nonetheless say, that such testing 

### 19 requires vetting of course. 

### 20 With respect to adults, the use of a 10-day 

### 21 viability test for adult bees, although it does present some 

### 22 interpretation challenges, nonetheless has a virtue of allowing 

### 23 for harmonization with ongoing efforts in Europe. 

### 24 **DR. DANIEL SCHLENK:**Dr. James? 

### 25 **DR. ROSALIND JAMES:**Yes please do them. I 

### 26 don't know how else to emphasize that the 48 hour, 96 hour tests

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1 are a great frustration to anybody working with bees and 

### 2 toxicity is how come EPA looks at such a short time period? I 

### 3 believe perhaps Dr. Pistorius can comment on - there are some 

### 4 standard OECD tests already available for the seven-day test. 

### 5 **MR. PISTORIUS:**Thank you. If I may answer 

### 6 directly, there is no real OECD test guideline available for the 

### 7 10 day test, but what is done and what is asked for in special 

### 8 cases with the concern of especially insecticides is that the 

### 9 OECD 213 is used and the protocol is used accordingly to OECD 

### 10 213, which means basically that all the test conditions, like 

### 11 how many bees per cage in all this is used accordingly, but the 

### 12 test is extended to 10 days. The only difference is that 

### 13 usually freshly emerged bees are taken, so this test could 

### 14 easily be done accordingly. 

### 15 **DR. DANIEL SCHLENK:**Okay. Dr. Pettis? 

### 16 **DR. JEFF PETTIS:**Just a bit of followup of 

### 17 when Jens said that in doing these cage studies, you often see 

### 18 that they use random age bees and for a longer test, it would 

### 19 not be appropriate. You would want to start the test with newly 

### 20 emerged bees. 

### 21 **DR. DANIEL SCHLENK:**And Dr. Ostiguy? 

### 22 **DR. NANCY OSTIGUY:**I concur with what's been 

### 23 said. 

### 24 **DR. DANIEL SCHLENK:**Okay. Any other Panel 

### 25 members? Okay. Mr. DeCant, any questions of clarification? 

### 26 Okay. Let's move on to C please and read that into the record.

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1 **MR. JOSEPH DECANT:**Joe DeCant, EFED, question 

### 2 10, part C - although a 10-day adult and seven-day larval 

### 3 toxicity tests have been proposed, please comment on whether 

### 4 alternative durations of pesticide exposure may be more 

### 5 appropriate for determining chronic toxicity for adult and 

### 6 larval bees at a Tier I screen. 

### 7 **DR. DANIEL SCHLENK:**And Dr. Berenbaum? 

### 8 **DR. MAY BERENBAUM:**A 10-day test of adult 

### 9 survival can be informative, but interpreting the biological 

### 10 meaning outside the social context remains challenging. Rather 

### 11 than an alternative duration of exposure, an alternative design, 

### 12 which we've all ready discussed, might be considered. Survival 

### 13 could be assessed in social context with the use of these cage 

### 14 brood or broodless bee cup tests, which have been used with some 

### 15 success in other contexts. 

### 16 As for the larval toxicity tests as we stated 

### 17 earlier, estimating the test duration through pupation rather 

### 18 than terminating it at an arbitrating interval would be most 

### 19 appropriate. 

### 20 **DR. DANIEL SCHLENK:**Dr. James? 

### 21 **DR. ROSALIND JAMES:**I concur with what was 

### 22 said. Just to clarify because she said it quickly, I mean there 

### 23 is an alternative way of doing brood tests. The standard tests 

### 24 are putting brood inside a container like a 48 well played and 

### 25 feeding them by hand and cleaning them, but you can also cage 

### 26 adult bees with some brood and allow the adult bees to care for

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1 the larvae, and you can still monitor the mortality of the 

### 2 larvae. 

### 3 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 4 **DR. JEFF PETTIS:**I concur with what's been 

### 5 said. 

### 6 **DR. DANIEL SCHLENK:**And Dr. Ostiguy? 

### 7 **DR. NANCY OSTIGUY:**I concur. 

### 8 **DR. DANIEL SCHLENK:**Any other comments? Mr. 

### 9 Pistorius? 

### 10 **MR. JENS PISTORIUS:**Thank you. Maybe chronic 

### 11 toxicity is a question of definition for the larva bees. It is 

### 12 for the larval test always the question of dosing. Is it acute 

### 13 dosing at one day of feeding or dosing on every day of feeding? 

### 14 We discussed this in the group and we were of the opinion that 

### 15 everyday should be - the pesticide should be added to food every 

### 16 day. 

### 17 **DR. DANIEL SCHLENK:**Other Panel members? 

### 18 Yeah, Dr. James? 

### 19 **DR. ROSALIND JAMES:**I'll just add to that a 

### 20 little bit. Essentially the larval feeding period is so short. 

### 21 A difference between acute and chronic, you know, it's hard. We 

### 22 have difficulty defining it. It's such a short period. It 

### 23 should be treated, perhaps like chronic, but it is still not a 

### 24 very long test. 

### 25 **DR. DANIEL SCHLENK:**Dr. Tarpy? 

### 26 **DR. DAVID TARPY:**This is Dave Tarpy. I

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1 think, to Dr. James's point, that that's why we avoided in 

### 2 question 8 beginning this morning, we're seeing a one-time 

### 3 application versus a continuous application type of language 

### 4 rather than acute and chronic, just to avoid that distinction of 

### 5 previous usage of that. 

### 6 **DR. DANIEL SCHLENK:**Anyone else? Dr. Hunt? 

### 7 8 **DR. GREG HUNT:**Well, I'll just reiterate what 

### 9 has been said before. The reason behind this is that it's more 

### 10 biologically relevant for the larvae because they're expected to 

### 11 be exposed to chronic doses. 

### 12 **DR. DANIEL SCHLENK:**Okay. Dr. Klaine? 

### 13 **DR. STEPHEN KLAINE:**Yeah, but I don't want to 

### 14 lose something that Dr. Delclos mentioned earlier, and that is 

### 15 timing of pesticide application. Really, the results are not 

### 16 really known. So, while for this particular test and a 

### 17 standardized test, it might be worthwhile to have pesticide dose 

### 18 to each day. Somebody needs to look at what happens when they 

### 19 only get in on, say day three or on day five, in terms of the 

### 20 development and ultimately the affect on things like pupation, 

### 21 et cetera. 

### 22 **DR. DANIEL SCHLENK:**Dr. James? 

### 23 **DR. ROSALIND JAMES:**But within the colony, 

### 24 you're going to have all different life stages going on at the 

### 25 same time, so within a colony - my name is Rosalind James - 

### 26 within a colony, you're going to have all the different life

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1 stages going on at once, and you'll also have all this 

### 2 trophallaxis and storage. It's just not - it's really rare that 

### 3 they're just going to one day get a dose in a colony and then 

### 4 it's all gone. 

### 5 **DR. STEPHEN KLAINE:**Steve Klaine - I agree 

### 6 with you about the different life stages going on at the same 

### 7 time, but in terms of from an exposure situation, it's not 

### 8 unrealistic to think that they would have a pulsed exposure 

### 9 where they might only see the pesticide one day or two days, 

### 10 depending on how that exposure occurs. 

### 11 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 12 **DR. MAY BERENBAUM:**Physiologically, it would 

### 13 be very useful and informative to know when pesticides exert 

### 14 their effects throughout the development, of course, their 

### 15 development of larvae. But in terms of the ecology of exposure, 

### 16 it's difficult to imagine how larvae would be subjected to acute 

### 17 exposures that weren't also at the same time horrifically 

### 18 catastrophic. So, you can imagine some kind of direct spray to 

### 19 the entire colony. But, larvae don't go anywhere and everything 

### 20 they eat is handed to them and it's already processed food. So, 

### 21 it's already been through several levels of - well it's been 

### 22 through several layers of bees actually. 

### 23 So, my bee biology colleagues, correct me if 

### 24 I'm wrong, but it's hard to conceive of a scenario of acute 

### 25 exposures that recur frequently at different developmental 

### 26 stages.

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1 **DR. DANIEL SCHLENK:**Dr. Ostiguy? 

### 2 **DR. NANCY OSTIGUY:**I think we might be 

### 3 talking about two different things here. Dr. Klaine, I think 

### 4 you're correct that it's a data gap that we don't know at what 

### 5 point we might have physiological changes that trigger in terms 

### 6 of Tier I or Tier II. I don't know that we need that kind of 

### 7 test done by an applicant. So, it's a data gap in terms of 

### 8 understanding some of the information that we might get from a 

### 9 Tier I or Tier II test, but it's not probably necessary to do 

### 10 during a Tier I or Tier II test. 

### 11 **DR. STEPHEN KLAINE:**This is Steve Klaine. 

### 12 You interpreted me correctly. Thanks. 

### 13 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 14 **DR. JEFF PETTIS:**I agree with Dr. Berenbaum 

### 15 that the majority of the food is processed and goes into larval 

### 16 development, but there is some direct pollen feeding and direct 

### 17 honey that is fed in late larval stage. So, there could be 

### 18 scenarios where you might see things coming into the hive that 

### 19 are then used directly. 

### 20 **DR. DANIEL SCHLENK:**It seems to me that 

### 21 you're kind of interested in windows of susceptibility during 

### 22 that developmental stage. So, when are those sort of critical 

### 23 periods of susceptibility that, you know, changes maybe an 

### 24 outcome that happens at that day, but wouldn't happen in another 

### 25 exposure period. I think that's - yeah. Any other comments or 

### 26 discussion? Okay. Let's move onto letter D. Sorry, I didn't

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1 ask for clarification questions, I'm sorry. Mr. DeCant? 

### 2 **MR. JOSEPH DECANT:**Yeah, Joe DeCant, EFED. 

### 3 Earlier in the discussion, Dr. James, I think you brought up 

### 4 the caged bee test, and that came up earlier in some of the 

### 5 Panels' comments. Is it a semi-field study that you are talking 

### 6 about or is this a laboratory based? 

### 7 **DR. DANIEL SCHLENK:**Yes, Dr. Tarpy? 

### 8 **DR. DAVID TARPY:**This is Dave Tarpy, NC 

### 9 State. These are in vitro studies, so they're all done in an 

### 10 incubator under controlled conditions, so these would be Tier I. 

### 11 However, I think what was just mentioned is by combining adults 

### 12 and larvae in vitro starts to blur Tier I and Tier II, but there 

### 13 are Tier I group bioassays that have been used successfully. 

### 14 That's what mostly we've been referring to up until now. 

### 15 **DR. DANIEL SCHLENK:**Yes, Dr. James? 

### 16 **DR. ROSALIND JAMES:**We had some discussions 

### 17 about how you would do the dosing. I mean, perhaps you could 

### 18 dose the larvae directly by hand and then have the adult bees 

### 19 take care of them so the exposure of the adult bees was minimal 

### 20 and you could make it a larval test. But, you're using a little 

### 21 more natural method for caring for the brood. When you do it 

### 22 yourself by hand, you maybe feed the bees every four hours or 

### 23 how often you feed them, and you have to clean out the wells. 

### 24 It's very labor intensive to do honey bee larval bioassays. 

### 25 It's a lot easier to let the nurse bees do this for you and they 

### 26 feed about every half hour, so the brood gets better care, and

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1 it would just be a little natural setting and this was one idea. 

### 2 So, it would be easier to conduct, I think, and the brood would 

### 3 get better care, so you would have lower control mortality. 

### 4 **MR. JOSEPH DECANT:**Joe DeCant, EFED. So, 

### 5 maybe just a clarification that this cage of brood study would 

### 6 be a replacement for the larval toxicity study to be able to 

### 7 take it through pupation? Or is it an alternative design that 

### 8 could be used, but the larval toxicity study as we have it in 

### 9 the Tier I screen would be an appropriate replacement given that 

### 10 we can take it through pupation. Once it goes through pupation, 

### 11 it would be sufficient in lieu of the cage brood study. 

### 12 **DR. DANIEL SCHLENK:**Dr. James? 

### 13 **DR. ROSALIND JAMES:**Sorry, Dr. Tarpy, are 

### 14 there standard methods published on this method? I am familiar 

### 15 with the method, but I'm not familiar with the literature on it. 

### 16 17 **DR. DAVID TARPY:**This is Dave Tarpy. I don't 

### 18 think there are standard methods on a caged brood study, but it 

### 19 is certainly, I think, feasible. But I don't think there is a 

### 20 standard think like the Aupinel paper on the hand rearing of 

### 21 larvae. 

### 22 **DR. DANIEL SCHLENK:**Yes, Dr. James? 

### 23 **DR. ROSALIND JAMES:**So, in lieu of that 

### 24 answer, my answer to you would be it would be an alternative 

### 25 method that could be used. 

### 26 **DR. DANIEL SCHLENK:**Dr. Ostiguy, did you have

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1 something to add? No. Dr. Berenbaum? 

### 2 **DR. MAY BERENBAUM:**And Aupinel offers the 

### 3 benefit of harmonization with European ongoing efforts. There 

### 4 are advantages to that, at least until method alternatives are 

### 5 developed. 

### 6 **DR. DANIEL SCHLENK:**Okay. Any other 

### 7 questions Mr. DeCant? Okay. Let's move onto D. 

### 8 **MR. JOSEPH DECANT:**Joe Decant, EFED, question 

### 9 10, subpart (d) - the White Paper identifies NOAEC as the 

### 10 chronic toxicity measurement endpoint. Please comment on the 

### 11 possible use of ECx values as a measure of chronic toxicity for 

### 12 use in RQ calculations. 

### 13 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 14 **DR. MAY BERENBAUM:**I must confess, there was 

### 15 a considerable amount of confusion about this question. If I'm 

### 16 not answer the question you're asking, I'm hoping that associate 

### 17 discussants will address it better. 

### 18 But the gist of our conversation generally is 

### 19 that the proposed regression based method for calculating ECx 

### 20 levels seems feasible if a sufficient number of concentrations 

### 21 can be tested to allow for confident estimates given the 

### 22 competence levels do drop at the high and low ends of dose 

### 23 response curves. 

### 24 Some discussion ensued on including a temporal 

### 25 component of chronic effects to provide a quantitative estimate, 

### 26 sort of a lethal time estimate like quantitative analysis of

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1 survivorship curves using Kaplan-Meier type survivorship as a 

### 2 way of estimating chronic impacts. I will leave it to the 

### 3 associate discussants to elaborate. 

### 4 **DR. DANIEL SCHLENK:**Dr. James? 

### 5 **DR. ROSALIND JAMES:**I like the use of 

### 6 something like a survivorship analysis that gives you a 

### 7 different kind of endpoint, but it doesn't give you and 

### 8 effective concentration because it gives you a time to effect 

### 9 for a given dose, and then has to be repeated at different 

### 10 doses. So, the calculation of the RQ wouldn't fit very well in 

### 11 the survivorship type of analysis. I don't have a better answer 

### 12 for you than that. 

### 13 **DR. DANIEL SCHLENK:**Dr. Ostiguy? 

### 14 **DR. NANCY OSTIGUY:**I concur with what's been 

### 15 said. 

### 16 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 17 **DR. JEFF PETTIS:**I have nothing to add. 

### 18 **DR. DANIEL SCHLENK:**Okay. Other Panel 

### 19 members? Yes, Dr. Fefferman? 

### 20 **DR. NINA FEFFERMAN:**So, I don't know if any 

### 21 of them would wind up appropriate, but there are a diversity of 

### 22 different measures for combining dose response and survival 

### 23 curves from the epidemiological human literature. So, that 

### 24 might be somewhere to look for methods that might fit with your 

### 25 requirements somewhat well, specifically in both cancer research 

### 26 for carcinogens, but also more specifically even in cancer for

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1 teratogens. The teratogen studies and time-tell spontaneous 

### 2 abortion or miscarriage is a place where I've seen a diversity 

### 3 of measures that might work really well for you. 

### 4 **DR. DANIEL SCHLENK:**Any other input? Okay. 

### 5 Oh, sorry, Mr. Pistorius? 

### 6 **MR. JENS PISTORIUS:**Maybe just a comment - in 

### 7 the coming OECD Guidance document, there is an attempt to 

### 8 integrate this chronic toxicity measurement endpoints and using 

### 9 ECx values and the use for risk assessment in trigger values. 

### 10 **DR. DANIEL SCHLENK:**Okay. If we could maybe 

### 11 reference that in the minutes, that would be good. Okay. Any 

### 12 other comments from the Panel on D? Okay. Mr. DeCant, any 

### 13 questions of clarification or Ms. Garber? Yeah? 

### 14 **MS. KRIS GARBER:**Yes please, Kris Garber. I 

### 15 think it might be a little helpful just to clarify the question 

### 16 a little bit, but I don't think we're quite getting what we're 

### 17 looking for, would that be all right? 

### 18 **DR. DANIEL SCHLENK:**Sure, yeah. 

### 19 **MS. KRIS GARBER:**Okay. So, just to put it in 

### 20 my own words, you know the endpoint that we traditionally rely 

### 21 upon to measure a chronic duration of exposure would be the no 

### 22 observed adverse effects level and you know, the design of that 

### 23 study. The study is designed in a particular way to derive that 

### 24 NOAEC concentration and, you know, there's been a lot of 

### 25 discussion in the scientific community about perhaps using an 

### 26 ECx value, and X would be whatever we decided to be biologically

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1 relevant. You see 10, you see 20 or EC50, of course, is 

### 2 relevant as well, and that would have, you know, a different 

### 3 study design as well. 

### 4 So, you know the specific endpoint that we 

### 5 would be going after for a chronic study is really relevant for 

### 6 considering the design of the chronic study. So, we're just 

### 7 curious on some thoughts for, you know, looking at effects to 

### 8 bees and the endpoints that you have all been discussion of the 

### 9 strengths and limitations of the NOAEC versus the EC whatever X 

### 10 is. I hope that helps. 

### 11 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 12 **DR. MAY BERENBAUM:**We, or at least I was a 

### 13 little confused about this question and how it relates to the 

### 14 next question, which seems to be setting the EC level, question 

### 15 C. So, maybe we can -- what percent effect - should I go on to 

### 16 C? E, right, sorry. I am myopic. I just wondered if we could 

### 17 meld these two, because what you're asking is where should EC be 

### 18 set and should we use EC? No. 

### 19 **DR. DANIEL SCHLENK:**No, she is not asking. I 

### 20 believe she's asking would you rather use a NOAEC or an ECx 

### 21 value. And which one, what are the benefits and you know, cost 

### 22 associated with those different measures essentially? Because 

### 23 one is a more deterministic sort of endpoint and the other one 

### 24 is much more based on the probabilistic curve. You set a value, 

### 25 which could be 5 percentile, 10 percentile, 20 percentile. I 

### 26 don't think she is asking what X should be, I think she's asking

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1 which method do you want to use. Would it be better to use, you 

### 2 know, a single point that's statistically significant above the 

### 3 note in terms of a single point value that you have in your data 

### 4 set or a derived value that you get from a probabilistic curve. 

### 5 That's what it sounds like. Yes, Mr. Pistorius? 

### 6 **MR. JENS PISTORIUS:**I hope this is slightly 

### 7 an answer or maybe helps. I assume now that for the chronic 

### 8 toxicity testing to determine the NOAEC, you would use the 

### 9 10-day tests. And then, it is the question how do you actually 

### 10 come closer to the dosing that you had testing. Do you go from 

### 11 an acute oral LD50 test identifying doses where you have no 

### 12 LD50? Then it will be a matter that you have to test quite a 

### 13 large number of doses to get a reliable level. Also, usually we 

### 14 know that with no-observed effect of concentrations and a low 

### 15 observed, that's why for other tests always the LD50 is used. 

### 16 The LD50 gives a lot higher certainty. So, that may be an 

### 17 argument to rather use the ECx values. 

### 18 **DR. DANIEL SCHLENK:**Yes, Dr. James? 

### 19 **DR. ROSALIND JAMES:**My brain is hurting on 

### 20 this question. But, the no-effect level will be more 

### 21 conservative, and maybe with some of these complex behavioral 

### 22 things, we want to trigger up higher tiered level. So, that 

### 23 would be an argument for the no-effect level. But, I don't know 

### 24 that we really know enough about the statistical variability 

### 25 that we're going to get out of these bioassays to really answer 

### 26 that question in terms what you're saying, unless you have

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1 experience with that. 

### 2 I really don't know how much variability 

### 3 you're going to have. Because sometimes like, you know, an EC10 

### 4 or an EC20 level may have a high amount of variability 

### 5 associated with it. It might be easier to draw a line at the 

### 6 no-effect level. You know, we are not always looking for the 

### 7 easy way out, but in some ways, triggering the Tier II for the 

### 8 behavioral effects might be a good thing to do if some effect is 

### 9 seen. 

### 10 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 11 **DR. MAY BERENBAUM:**The one advantage of EC 

### 12 versus no-adverse effect level is that it's always an open 

### 13 question just to why no effects have been observed. So, an EC 

### 14 measurement at least is quantifying a known response to the 

### 15 degree that that's reassuring and repeatable that could be an 

### 16 advantage for using EC rather than no observable effect. 

### 17 **DR. DANIEL SCHLENK:**Dr. Klaine? 

### 18 **DR. STEPHEN KLAINE:**Yeah, let me weight on 

### 19 this. As you indicated before, using the NOAEC has a problem in 

### 20 that fact that it's heavily dependent on the on experimental 

### 21 design. So, I'm not sure that the issue with bees has any 

### 22 relevance to which one you use. I think that the better choice 

### 23 would be the EC number because its derived in a more realistic 

### 24 or more probabilistic manner. But, I'm really pleased that you 

### 25 brought up the issue of variability because, Dr. James, because 

### 26 I think that's the real driver here. We all use EC50s because

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1 it's the most accurate. However, if you have the ability to use 

### 2 a higher end, you then drive the uncertainty regarding an EC5. 

### 3 You decrease that uncertainty. So, it may make it more 

### 4 realistic with this organism to actually generate EC5s than with 

### 5 classic toxicological organisms where it's impossible to get the 

### 6 N above the 3 or 4 or 5. So. 

### 7 **DR. DANIEL SCHLENK:**Mr. Pistorius? 

### 8 **MR. JENS PISTORIUS:**I think that in any case, 

### 9 it is an old problem that both the ECx values and the NOAEC 

### 10 levels are reported from those studies. So, I just question 

### 11 which one to use later on, but you can always, from those 

### 12 studies, get both values. 

### 13 **DR. DANIEL SCHLENK:**Dr. Ostiguy? 

### 14 **DR. NANCY OSTIGUY:**If there is a question 

### 15 about a low sample size, then my preference is the NOAEC. But 

### 16 for a large sample size, setting a low EC value, that would 

### 17 absolutely be preferable. So, the larger sample size would 

### 18 certainly produce better data. 

### 19 **DR. DANIEL SCHLENK:**Any other comments? Ms. 

### 20 Garber, does that answer your question? Oh, Mr. Sappington? 

### 21 **MR. KEITH SAPPINGTON:**Keith Sappington. I 

### 22 think, yeah, this question is difficult. The NOAEC versus the 

### 23 hypothesis testing, i.e. NOAEC versus point estimation ECx issue 

### 24 has been around for decades. So, that issue is generic. I 

### 25 think it would be helpful - and there are advantages and 

### 26 disadvantages to each one, and there have been a number of

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1 papers published on that. 

### 2 In my mind, it would be helpful to know if 

### 3 given the biology of the honey bee or non-Apis bees, but given 

### 4 the social nature of at least honey bees, might there be an 

### 5 Apis-specific reason for favoring one over the other. So, that 

### 6 is kind of perhaps where we're coming from, because this is an 

### 7 issue for all tox tests. 

### 8 **DR. DANIEL SCHLENK:**Anybody want to comment 

### 9 on that? Yes, Dr. Hunt? 

### 10 **DR. GREG HUNT:**I don't know much about 

### 11 testing pesticides, but you can expect with honey bees to have 

### 12 with this kinds of tests, kind of small sample size and hive 

### 13 variability. So, that should factor into your thinking. 

### 14 **DR. DANIEL SCHLENK:**Okay. Dr. Berenbaum, 

### 15 since you're kind of the lead discussant, do you need any more 

### 16 input in terms of how to answer that? Or are you good? 

### 17 **DR. MAY BERENBAUM:**I don't think so. 

### 18 **DR. DANIEL SCHLENK:**Okay. Dr. Klaine? 

### 19 **DR. STEPHEN KLAINE:**Not to beat this, but Dr. 

### 20 Hunt, why do we have to have small sample size? 

### 21 **DR. GREG HUNT:**Well, you don't have to have 

### 22 small sample size, of course, given enough resources, but it's 

### 23 difficult to get a lot of incubators and a lot of cages and to 

### 24 the practicality of finding emerging brood, which you need to 

### 25 emerge in an incubator and collect the bees, get it all going at 

### 26 the same time before, you know, they start dying from lack of

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1 food and there are practical constraints on it. And we see in 

### 2 cage studies hive variability without treatment in terms of 

### 3 mortality. 

### 4 **DR. DANIEL SCHLENK:**Mr. Pistorius? 

### 5 **MR. JENS PISTORIUS:**One remark, I just saw 

### 6 that in the OECD Guidance document, which would be out for 

### 7 public commenting next week, I think, there is an approach 

### 8 which, and also a small discussion about which could be used and 

### 9 why. So, just as a recommendation maybe. 

### 10 **DR. DANIEL SCHLENK:**We will include that 

### 11 reference as a document to come, I guess, as you meant. 

### 12 **MR. JENS PISTORIUS:**It will be out for public 

### 13 consultation, so everybody can vote on this by the end of next 

### 14 week as a visual document. 

### 15 **DR. DANIEL SCHLENK:**Great. Okay. You have 

### 16 what you need now? Let's move on into letter E. 

### 17 **MR. JOSEPH DECANT:**Joe DeCant, question 10, 

### 18 subpart (e) - please provide comments on what percent effect 

### 19 would be considered a relevant measure of chronic toxicity for 

### 20 individual bees given the potential compensatory effects which 

### 21 honey bee colonies may exhibit relative to the effects of 

### 22 pesticides. 

### 23 **DR. DANIEL SCHLENK:**And Dr. Berenbaum? 

### 24 **DR. MAY BERENBAUM:**Compensatory responses are 

### 25 essentially a society-level phenomenon, so, inherently or maybe 

### 26 better suited for Tier II assessment. Recruitment of precocious

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1 foragers provides a short-term solution to colonies but may 

### 2 exert long-term costs on colony health. There is one study 

### 3 cited in the White Paper, Khoury et al. 2011, that estimates a 

### 4 reduction of worker longevity of 2.8 days corresponds to a loss 

### 5 of 0.35 bees per day. 

### 6 With that said, then aiming for an EC level 

### 7 that takes that number into account may be an approach for 

### 8 addressing the compensatory behavior. How that translates, 

### 9 we're not sure, but the recommendation is to be conservative. 

### 10 So, if it's not 35 percent, because its 0.35 bees per day, but 

### 11 in general, based on discussions yesterday, it seemed that a 

### 12 level of EC10 or EC20 might be conservative and attainable under 

### 13 these experimental conditions. 

### 14 What we think the question is at what point 

### 15 will worker losses send the colonies into the irretrievable 

### 16 death spiral. And again, there was a general subjective 

### 17 impression that 35 percent is not sustainable documenting that 

### 18 with literature was difficult. So, there is basically one 

### 19 study, possibly two studies. That was the extent of our 

### 20 discussion. 

### 21 **DR. DANIEL SCHLENK:**Dr. James? 

### 22 **DR. ROSALIND JAMES:**Maybe some of the 

### 23 modeling that we'll discuss later today or tomorrow will help 

### 24 answer this question? 

### 25 **DR. DANIEL SCHLENK:**Dr. Ostiguy? 

### 26 **DR. NANCY OSTIGUY:**I concur with the

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1 comments. 

### 2 **DR. DANIEL SCHLENK:**And Dr. Pettis? 

### 3 **DR. JEFF PETTIS:**Just kind of reiterating 

### 4 what Dr. Berenbaum said that there was a general feeling that 

### 5 somewhere around 30 percent loss of foragers might be a tipping 

### 6 point, if you will, on the colony level and then maybe something 

### 7 slightly more conservative than the 20 percent realm might be 

### 8 reasonable, but we don't have good data to back that up. And 

### 9 yes, maybe a modeling approach would help. 

### 10 **DR. DANIEL SCHLENK:**Okay. Any other - yes, 

### 11 Dr. Hunt? 

### 12 **DR. GREG HUNT:**Just to clarify, and I'm not 

### 13 sure, but I think that we're talking about like a 30 percent 

### 14 ongoing loss. Correct me if I'm wrong, because a one-time loss, 

### 15 they could recover from that. 

### 16 **DR. DANIEL SCHLENK:**So, an ongoing loss of 30 

### 17 percent sounds like the estimate, right? Dr. Tarpy? 

### 18 **DR. DAVID TARPY:**This is David Tarpy. I 

### 19 should also note that in a full colony, we're still in Tier I 

### 20 here, but in a full colony situation that that's very 

### 21 environmentally contextual, so that different times of year, 

### 22 colonies are more or less susceptible. So in the fall when 

### 23 they're populations are naturally declining, a smaller 

### 24 proportion of bee loss would probably have a much larger impact 

### 25 versus in the spring when their colonies are rapidly increasing, 

### 26 presumably more likely to be able to absorb that type of loss.

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1 But again, modeling approaches I think would be very informative 

### 2 here. 

### 3 **DR. DANIEL SCHLENK:**Dr. Fefferman? 

### 4 **DR. NINA FEFFERMAN:**So, just quickly - my 

### 5 reason for our needs on that if that the ramp up is actually 

### 6 supporting the continued ramp up, so that if you decrease too 

### 7 much during that increased growth, what you've done has lost the 

### 8 capacity to support increased queen fecundity. So, you're 

### 9 right, I want to model it. 

### 10 **DR. DANIEL SCHLENK:**Mr. Pistorius? 

### 11 **MR. JENS PISTORIUS:**I would just like to 

### 12 point out that in my opinion it is not appropriate to state 30 

### 13 percent mortality no matter at what time of the season is 

### 14 acceptable in this discussion here, and also later on I think, 

### 15 30 percent mortality even of foragers could be compensable by 

### 16 the colony, but this may be my perspective as a risk assessor is 

### 17 not acceptable. 

### 18 **DR. DANIEL SCHLENK:**Yes, Dr. Ostiguy? 

### 19 **DR. NANCY OSTIGUY:**Actually, I think what our 

### 20 discussion was that if it hit 30 percent, the colony was at the 

### 21 point of no recovery. So, the level of acceptable loss level 

### 22 has to be set less than that. The problem is that we don't have 

### 23 data to say where that should be. Our sense was somewhere 

### 24 around 20 percent. 

### 25 **DR. DANIEL SCHLENK:**Yes, Mr. Pistorius? 

### 26 **MR. JENS PISTORIUS:**I think that even in some

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1 cases, colony an recover if the damage is more than 30 percent 

### 2 because we've had some incidents where larger percentages were 

### 3 damaged. But on the other hand, I'm not quite sure if this is 

### 4 the issue of the questions that we are currently discussion. I 

### 5 don't know if we're supposed to intend to extend this discussion 

### 6 at this moment. 

### 7 **DR. DANIEL SCHLENK:**Dr. Ostiguy? 

### 8 **DR. NANCY OSTIGUY:**I think what we were 

### 9 talking about was a continuous 30 percent loss, not a one-time 

### 10 loss, because certainly colonies can recovery from a significant 

### 11 loss depending on environmental conditions. 

### 12 **MR. JENS PISTORIUS:**I agree, and I think the 

### 13 difficulty was that we have just been mingling laboratory issues 

### 14 and realistic world conditions, basically in the discussions. 

### 15 But I agree to what Nancy Ostiguy has said. 

### 16 **DR. DANIEL SCHLENK:**Okay. Any other 

### 17 comments? Okay. Mr. DeCant, do you have what you need for that 

### 18 question? Okay. All right, let's move on to the next 

### 19 subsection and read that into the record. 

### 20 **MR. JOSEPH DECANT:**Joe DeCant, EFED, question 

### 21 10, subpart (f) - although the White Paper identifies some 

### 22 measurement endpoints for assessing chronic toxicity to 

### 23 individual bees such as survival, please comment on other 

### 24 endpoints to consider in chronic toxicity studies which the 

### 25 Panel believes are important and the associated study design 

### 26 elements.

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1 **DR. DANIEL SCHLENK:**And Dr. Berenbaum? 

### 2 **DR. MAY BERENBAUM:**As Dr. Tarpy mentioned in 

### 3 the response to question 8, other endpoints to consider in 

### 4 chronic toxicity tests may include behavioral impacts, learning 

### 5 assays, automated recording of motility and behavioral 

### 6 interactions and the like, although these haven't been vetted 

### 7 extensively in the context of chronic toxicity. 

### 8 Future endpoints could include gene expression 

### 9 relative to detoxification and/or antioxidant status, 

### 10 immunocompetence senescence traits and the like. Such surrogate 

### 11 biomarkers are widely used in human risk assessment. They 

### 12 haven't been validated for bees, but in many ways, they are more 

### 13 appropriate given the social nature of the organism. With that 

### 14 said, we don't have any - at least yesterday, discuss any 

### 15 guidelines known for identifying biomarkers or validating them. 

### 16 17 **DR. DANIEL SCHLENK:**Okay. Dr. James? 

### 18 **DR. ROSALIND JAMES:**I think we answered this 

### 19 in 8B when we talked, the similar question was proposed for 

### 20 acute toxicity. Since we were recommending just one test, we 

### 21 really just answered the question already in 8B, time to 

### 22 pupation and larval development time, success of pupation, that 

### 23 sort of thing. 

### 24 **DR. DANIEL SCHLENK:**And Dr. Ostiguy? 

### 25 **DR. NANCY OSTIGUY:**One thing that we might, 

### 26 that would be a future possibility, there has been work done

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1 looking at colony level ability to respond to health threats, 

### 2 would be one way to describe it. And these are responses to 

### 3 specifically, pathogens is where the work has been done that 

### 4 occur at a colony level but not at an individual level. And, 

### 5 that would then be potentially a future - well hopefully a 

### 6 future test, but there's a lot of data that needs to be 

### 7 collected to make that at all viable. 

### 8 **DR. DANIEL SCHLENK:**Sounds like the colony 

### 9 level effects will be discussed later on, yeah? Okay. And Dr. 

### 10 Pettis? 

### 11 **DR. JEFF PETTIS:**I concur with what has been 

### 12 said. 

### 13 **DR. DANIEL SCHLENK:**Dr. Fefferman? 

### 14 **DR. NINA FEFFERMAN:**As one potential 

### 15 mechanism for sublethal kinds of effects like this one that we 

### 16 haven't discussed so much is response to temporal task 

### 17 switching. So, most bees have an age-based polyethism of what 

### 18 they'll do primarily, but in terms of things like health, not 

### 19 just health threats, but things like if there's a point source 

### 20 of heat, the recognition of individual bees to go and act as 

### 21 their coolant batteries themselves, heat shielding, response to 

### 22 heat shielding, response to solicitation from nurse bees and the 

### 23 lack of nursing bees from older bees, switching back to nursing 

### 24 with that stimulus. Those kinds of task switching behaviors 

### 25 could be critical over chronic toxicity. 

### 26 So, all of the these questions sort of boil

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1 down into my mind into how stoned does the average New Yorker 

### 2 have to be for the busses to stop running? And that's really 

### 3 hard to measure locally. So, but I haven't heard anyone talk so 

### 4 much about the communication and solicitation response aspects 

### 5 of colony organization. I want to put that out there as one 

### 6 thing to try and measure as we look at this. 

### 7 **DR. DANIEL SCHLENK:**Mr. Pistorius? 

### 8 **MR. JENS PISTORIUS:**In addition to what Dr. 

### 9 James has said, on the in vitro studies, I just want to refer 

### 10 the comment I made before for the acute contact and oral 

### 11 toxicity studies that in this case. Also the behavioral aspects 

### 12 should be noted and this might, even if not really, a trigger 

### 13 can be recommended on what portion of bees we see those effects 

### 14 that the risk assessments needs to have the flexibility to 

### 15 trigger other tests on this information of sublethal behavioral 

### 16 abnormalities, especially because when you think that a chronic 

### 17 toxicity test is aiming at doses going below the LD50 and its 

### 18 behavioral abnormalities persist, then it might trigger further 

### 19 concern. The risk assessment needs an option to go to a higher 

### 20 tier test or to other tests as needed. 

### 21 **DR. DANIEL SCHLENK:**Okay. Any other input? 

### 22 Yes, Dr. Potter? 

### 23 **DR. THOMAS POTTER:**Yes, this is Tom Potter. 

### 24 We had a few side bar discussions here at the last break, 

### 25 discussing the possibility of using respiration measurements as 

### 26 index of stress. And, I would be interested to hear some

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1 comments from the rest of the Panel on whether or not that would 

### 2 be a useful approach. 

### 3 **DR. DANIEL SCHLENK:**Anybody want to weigh in 

### 4 on that one? Dr. James? 

### 5 **DR. ROSALIND JAMES:**This is a little remote, 

### 6 but my first thought is that there really aren't any 

### 7 standardized sort of methods for relating that to anything, 

### 8 except there is a fair amount of work done with respiration 

### 9 rates and diapause in the dormant solitary bees. The honey bee. 

### 10 **DR. DANIEL SCHLENK:**Can you - do you want to 

### 11 - can you get that on record please? 

### 12 **DR. NINA FEFFERMAN:**Sorry, Nina Fefferman. 

### 13 Just that there's a lot on colony level respiration needs and 

### 14 stress and success of colonies in ants, but I don't know of 

### 15 anything in bees. 

### 16 **DR. DANIEL SCHLENK:**Yes, Dr. James? 

### 17 **DR. ROSALIND JAMES:**At the colony level, so a 

### 18 Tier II type of test? 

### 19 **DR. NINA FEFFERMAN:**Actually both individual, 

### 20 but those tend to be challenged type individual studies where 

### 21 they take an ant and they either an antimate pheromone signal 

### 22 that signals help, there's a bird or something like that, and 

### 23 then there's sort of what you would call psychological stress 

### 24 for individual ants, but then yeah, most of the stuff is really 

### 25 quite good for regulating stress and colony impact at the Tier 

### 26 II level, yeah.

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1 **DR. DANIEL SCHLENK:**Yes, Dr. Hunt? 

### 2 **DR. GREG HUNT:**Well, there are studies in 

### 3 looking at the flight metabolic rate of honey bees, which have 

### 4 the highest mass flight metabolic rate of anything, even humming 

### 5 birds. So, I mean that could be investigated, and it being a 

### 6 physiological test, it could be a good stress test. 

### 7 **DR. DANIEL SCHLENK:**Yes, Dr. Pettis? 

### 8 **DR. JEFF PETTIS:**I was going to follow up on 

### 9 what Greg said that we have these treadmills that you can 

### 10 actually just glue the bee to and force it to fly and look 

### 11 energetic, so it might be - it hasn't been fully developed, but 

### 12 it might be a way to go about it. 

### 13 **DR. DANIEL SCHLENK:**So, a flight tunnel, 

### 14 right? Okay. So, it sounds like there are some other endpoints 

### 15 that are still in development so to speak that could be, you 

### 16 know, proposed - physiological biomarkers perhaps, if you want 

### 17 to put it into a category I guess. Okay. Any other comments on 

### 18 other endpoints? Okay. I think we got barely enough brain left 

### 19 for one more question before lunch here if we can squeeze in 

### 20 this last letter here. So, Mr. DeCant, do you want to read that 

### 21 into. 

### 22 **MR. JOSEPH DECANT:**Joe DeCant, question 10, 

### 23 subpart (g), section 4.1.3 of the White Paper discusses the 

### 24 uncertainties associated with developing risk assessments based 

### 25 on studies of sublethal effects when sufficient linkages have 

### 26 not been developed to understand how the sublethal endpoints may

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1 be quantitatively related to typical assessment endpoints, that 

### 2 is growth, impaired survival, and reproduction, at the whole 

### 3 colony level. Please comment on the proposal to use data on 

### 4 sublethal endpoints to qualitatively, that is, no Risk Quotients 

### 5 are derived to characterize effects and risk until sufficient 

### 6 linkages between these endpoints and the defined assessment 

### 7 endpoints have been developed as in the Adverse Outcome 

### 8 Pathways. 

### 9 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 10 **DR. MAY BERENBAUM:**The social environment 

### 11 complicates the process of identifying reliable, reproducible 

### 12 and relevant sublethal endpoints. But given the complexities, 

### 13 sublethal endpoint use is conservative and is very important for 

### 14 providing insight into super-organism responses and they are 

### 15 critical triggers for signaling a need for moving onto Tier II 

### 16 or higher tiers. So, we would be considered generally advisable 

### 17 not to discount or dismiss sublethal endpoints. 

### 18 An important priority for future research 

### 19 though, would be developing assessments for neurotoxicity 

### 20 symptoms. Although direct links to colony health may not yet be 

### 21 fully understood, they're indicative of biological activity of 

### 22 chemicals and therefore appropriate candidates for further 

### 23 investigation at the colony level. 

### 24 So, the results of these types of studies can 

### 25 demonstrate a need for additional quantitative tests at Tier II. 

### 26 Eventually, links have to be made, but given the state of

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1 knowledge at the present time, sublethal endpoints are really 

### 2 critical indicators for biological activity that may be 

### 3 difficult to assess using traditional toxicological measures 

### 4 designed for organisms that are not so dependent on a social 

### 5 environment. 

### 6 **DR. DANIEL SCHLENK:**Okay. Dr. James? 

### 7 **DR. ROSALIND JAMES:**I concur, but I will add 

### 8 in defense of the nonsocial bees, they have complex behaviors 

### 9 also in nest building and so on. It would be applicable to the 

### 10 non-Apis bees as well. 

### 11 **DR. DANIEL SCHLENK:**Dr. Ostiguy? 

### 12 **DR. NANCY OSTIGUY:**Just in case this matters, 

### 13 there is not a 4.1.3, so I think it's a 4.1.2.1. So, we think 

### 14 we're answering the correct one, but let us know if we're not. 

### 15 **DR. DANIEL SCHLENK:**Okay. 

### 16 **DR. MAY BERENBAUM:**This is May Berenbaum. 

### 17 4.1.2.1.2 is the section. 

### 18 **DR. DANIEL SCHLENK:**Yeah. I got that one. 

### 19 Okay. Dr. Pettis? 

### 20 **DR. JEFF PETTIS:**I concur. 

### 21 **DR. DANIEL SCHLENK:**That we're on the right 

### 22 number? 

### 23 **DR. JEFF PETTIS:**We actually spent quite a 

### 24 bit of time trying to find the 4.3. Are we missing something, 

### 25 but no we haven't. 

### 26 **DR. DANIEL SCHLENK:**Just a challenge, it was

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1 a challenge. Yes? Dr. Fefferman? 

### 2 **DR. NINA FEFFERMAN:**Sorry, in all 

### 3 seriousness, this is yet another one that I think we'll end up 

### 4 discussing the context for later. But really anytime that these 

### 5 questions relate to how to scale unknown relationships linking 

### 6 individual level sublethal effects, or lethal effects frankly, 

### 7 to the colony level compromise, that's a place where modeling 

### 8 actually may be able to help. Even working backwards toward a 

### 9 Tier I type study to recommend what kinds of sublethal behaviors 

### 10 could be used as proximate measures in individuals that would 

### 11 then scale up to call any compromise, but we'll talk about that 

### 12 more later. 

### 13 **DR. DANIEL SCHLENK:**Any other comments? So, 

### 14 it sounds like at least for adverse outcome pathways, 

### 15 neurotoxicity would be at least one sort of pathway to pursue 

### 16 it. I'm interested in the Panel's response to the beekeeper 

### 17 that had the eviscerated males that were lying around. Would 

### 18 that indicate maybe and endocrine pathway would be another sort 

### 19 of target that would be something to explore. Yes, Dr. Hunt? 

### 20 **DR. GREG HUNT:**I don't really know why drones 

### 21 fall dead with their genitalia everted, but I think it has 

### 22 something to do with general stress. I'll leave it there. 

### 23 **DR. DANIEL SCHLENK:**Yes, Dr. Tarpy? 

### 24 **DR. DAVID TARPY:**This is David Tarpy. This 

### 25 is pure speculation, but because the kind of default situation 

### 26 for drones is to turn themselves inside out to evert their

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1 genitalia to mate, but it's some of the lower ganglion that 

### 2 inhibit that, that it actually suggest a potential neural 

### 3 pathway for that particular report. But, that's again, just 

### 4 speculation. 

### 5 **DR. DANIEL SCHLENK:**All right. Any other 

### 6 comments? Mr. DeCant, do you have questions of clarification? 

### 7 **MR. JOSEPH DECANT:**Joe DeCant, EFED. I just 

### 8 want to sort of make sure that I understood the panel correctly, 

### 9 that the sublethal endpoints, some of them are still being 

### 10 developed. The assays are either still in development or we're 

### 11 still waiting on the quantitative linkage between the sublethal 

### 12 endpoints and some of the colony level effects. Given that 

### 13 though, these sublethal endpoints are important and that while 

### 14 we may not use them quantitatively, the panel would agree that 

### 15 we should use them qualitatively and perhaps use them to also 

### 16 trigger higher tier studies. 

### 17 **DR. DANIEL SCHLENK:**I see most heads nodding 

### 18 on that one. So, it seems like a general agreement. Okay. All 

### 19 right, I think we're sufficiently toasted. So, let's take a 

### 20 lunch break and come back at 12:45. 

### 21 (WHEREUPON lunch was taken) 

### 22 All right, if you would please take your 

### 23 seats. Okay. Let me just go to the Agency. Do you have any 

### 24 other clarifying questions that you have before we start with 

### 25 question 11 on any of the previous questions? We're good? 

### 26 Great. Okay. Let's go ahead and read 11A into the record if

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1 it's up there. 

### 2 **MR. JOSEPH DECANT:**Joe DeCant, EFED, question 

### 3 11, subpart (a) - although colonies are typically confined to 

### 4 enclosures for Tier II studies and these enclosures can limit 

### 5 the environmental realism of the study conditions, tunnel 

### 6 studies provide an opportunity to collect colony-level effects 

### 7 and potentially exposure information. Please comment on the 

### 8 relative strengths, that is foraging activity by adult worker 

### 9 bees is limited to treated crop; trophyllaxis enabled and 

### 10 limitations such as limited study duration, smaller colony sizes 

### 11 and reduced forage area of these methods. 

### 12 **DR. DANIEL SCHLENK:**Lead discussant for 

### 13 that's Dr. Tarpy. 

### 14 **DR. DAVID TARPY:**This is David Tarpy, NC 

### 15 State, good afternoon everybody. I want to preface the comments 

### 16 in talking about Tier II and other studies by kind of briefly 

### 17 reiterating the point of linking this to our discussion this 

### 18 morning about Tier I bioassays, and how when we're looking at 

### 19 social insects such as honey bees that it starts to blur the 

### 20 lines very much. So, that simple LD50 mortality measurements of 

### 21 individual adult worker honey bees doesn't actually really 

### 22 capture the myriad of different exposure pathways and other 

### 23 effects of social insects. So, this to me is really the 

### 24 fundamental question that faces the panel and is driving a lot 

### 25 of our discussions for this White Paper. 

### 26 So, one means of addressing that obviously is

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1 to incorporate a lot of these other bioassays that we discussed 

### 2 in question 8 and others to help bridge this gap, but it still 

### 3 doesn't really fully serve as a proxy for colony-level 

### 4 phenotype. And again, this is because honey bee colonies are 

### 5 like super organisms. So, in that way, biologically the basal 

### 6 unit is that entire colony. It's very hard to test these 

### 7 systems in a reductionistic approach. 

### 8 The kind of analogous or an analogy to this is 

### 9 to study individual cell cultures of say a rat and looking at 

### 10 LD50s of individual cells of that cell culture and then trying 

### 11 to extrapolate that to the entire rat. So, the way that it's 

### 12 actually done is to study individual rats and looking at LD50s 

### 13 of different compounds of whole rats. This is the same that is 

### 14 true for entire colonies. 

### 15 So, in some ways, it's inescapable to have to 

### 16 test entire colonies to find full phenotypic effects at that 

### 17 level. Not to say that Tier I screens of individual bees or 

### 18 brood are not informative, it's just that they are not perfect 

### 19 proxies or surrogates for higher levels at the colony level. 

### 20 So with all that said and with all that in 

### 21 mind, the question asked to identify some strengths and 

### 22 weaknesses of Tier II studies, and obviously some to strengths 

### 23 or because it's measuring at in vivo in this super organism 

### 24 context, it does better enable and capture a lot of those 

### 25 complex social interactions that are going on that result in 

### 26 full fledge colony phenotype.

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1 The limitations, many of which are kind of 

### 2 logistical in that there is difficulty in getting large sample 

### 3 size when you're dealing with entire colonies. It's just a 

### 4 realistic problem with trying to get sufficient number of 

### 5 colonies, especially because they're so highly variable social 

### 6 insects. One of their keys to their ecological success is their 

### 7 huge variability and their ability to adapt different 

### 8 environments. So, there's a lot of variation across colonies. 

### 9 So, trying to account for that requires higher sample sizes and 

### 10 that's obviously a real logistical constraint. Again, 

### 11 automation of data collection and means to be able to streamline 

### 12 that would be incredibly helpful, but I don't think the 

### 13 technology is quite there yet. 

### 14 In doing so, it's always helpful - or the 

### 15 Panels feel that the type Tier II studies, it's very helpful to 

### 16 contextualize a lot of these colony level phenotypes by always 

### 17 having adequate control, either positive and/or negative control 

### 18 groups to relativize it. That helps be able to place the 

### 19 findings into the proper context. It's hard to have kind of 

### 20 universal measurements across these different studies because 

### 21 they're done different times of the year under very different 

### 22 conditions, environmental conditions. So, doing to contextually 

### 23 is very helpful. 

### 24 We can discuss this in part B and in other 

### 25 places, but we do also want to throw out the notion that the two 

### 26 types of type II studies that were discussed in the White Paper,

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1 one was enclosed foraging arenas in cages and then others where 

### 2 they were on a treated target crop and other where they were 

### 3 free flying, but they were being actively fed through 

### 4 supplemental feed these different compounds in question that I 

### 5 think that the former approach might be more amenable to other 

### 6 social insects, particularly bumble bees. 

### 7 Honey bees are notoriously difficult to keep 

### 8 in small enclosed areas. Their foragers always want to be 

### 9 trying to go outwards, further outwards, and that's why honey 

### 10 bees are really not used in greenhouse pollination, for example. 

### 11 They're just not as amenable to that type of condition. Bumble 

### 12 bees, however, which are primitively used social and not highly 

### 13 used social, but they still live in colonies. They tend to be 

### 14 much more amenable to those types of systems. 

### 15 For the free-flying type of approach however, 

### 16 honey bee colonies do have some strengths and would probably be 

### 17 more amenable to that type of Tier II approach. 

### 18 **DR. DANIEL SCHLENK:**Okay. Thanks. Dr. 

### 19 Ostiguy? 

### 20 **DR. NANCY OSTIGUY:**Our concern about Tier I 

### 21 studies that Dr. Tarpy was mentioning because of honey bees 

### 22 being a social insect may be partially addressed when we talk 

### 23 about question 14 and I'm going to probably use the wrong word, 

### 24 Nina, but meta-models. So, the higher level of modeling so we 

### 25 can select which of the Tier I studies might actually provide us 

### 26 with the best information about what's going to happen then at

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1 the colony level. 

### 2 So, there may be a way for us to address that 

### 3 social insect issue that in some ways is better studied 

### 4 empirically or with hard data with Tier II and Tier III, Tier I 

### 5 still may provide us with some useful information once we have a 

### 6 better idea of which of those Tier I pieces of data that we 

### 7 might collect, actually would trigger colony-level problems. 

### 8 **DR. DANIEL SCHLENK:**Thanks. And Dr. Pettis? 

### 9 10 **DR. JEFF PETTIS:**Just a bit to add. There's 

### 11 a question about colony size in some of these Tier II and Tier 

### 12 III studies and how small colonies might not be representative 

### 13 of large colonies. I think if small colonies are properly set 

### 14 up, even if their nucleus colonies, the can be somewhat 

### 15 representative of whole colonies. You actually might - using 

### 16 small colonies, you actually might see an effect, which you 

### 17 wouldn't see in larger colonies. So, I just wanted to get that 

### 18 point in. I concur with the other comments. 

### 19 **DR. DANIEL SCHLENK:**Other Panelists? Yes, 

### 20 Dr. Berenbaum? 

### 21 **DR. MAY BERENBAUM:**Just one caveat about 

### 22 supplemental feeding, there are among these types of studies 

### 23 posed in the literature, there is supplemental feeding and 

### 24 there's force feeding, and it's important to emphasize that 

### 25 honey bee behavior is incredibly sophisticated, particularly 

### 26 with respect to food selection and food evaluation. So, suffice

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1 to say that not all supplemental feeding studies may be 

### 2 equivalently useful. 

### 3 **DR. DANIEL SCHLENK:**Okay. Mr. Pistorius? 

### 4 **MR. JENS PISTORIUS:**I would like to go into 

### 5 detail on the usefulness of different semi-field studies 

### 6 described here, those feeding studies later on. Let me say this 

### 7 all ready, I think it is absolutely -- it has to be refused to 

### 8 use such feeding studies. They are - for me personally, they 

### 9 are intermediate between Tier I and Tier II, but they're not 

### 10 really a Tier II test. I would give the justification for this 

### 11 rather on the point D when we look at different studies. 

### 12 So, now for me, I'm talking about the OECD 75 

### 13 and EPPO 170, which I think are very appropriate to with the 

### 14 worst case enclosure to assess the acute effects on bees. Of 

### 15 course, there are some limitations we know in the confinement. 

### 16 For instance, there may be a natural reduction of the brood 

### 17 because the bees don't want to breed as much under natural 

### 18 conditions, that in effect what you can eradicate by appropriate 

### 19 replications. But I think that, especially the EPPO 170 and 

### 20 OECD 75 are the ones to go for as a Tier II test. 

### 21 You have acute and you've got oral and contact 

### 22 exposure combined. What has come out of the comments that we 

### 23 basically want to have a look at all the different exposure 

### 24 routes and effect on the bee colony. And small colonies, I 

### 25 agree with Jeff, may be used to interpret the effects on 

### 26 colonies to some extent. In some cases, it might be a necessary

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1 step to go to Tier III, but it gives you a clear indication, 

### 2 especially the OECD 75 method, I think, is very reliable. For 

### 3 one, it builds on the EPPO 170, but specifies the brood 

### 4 assessment. 

### 5 This in general allows to have really worst 

### 6 case exposure of bees, of different castes of bees. You have 

### 7 some certain limitations that you have to supply some food, but 

### 8 that is always the case with every method except one feeding 

### 9 method where you could reduce the amount to almost zero, but you 

### 10 have to apply a little bit of food. Of course, there is a 

### 11 little uncertainty, but I think in general, they are very 

### 12 appropriate studies, but only the OECD 75 and the EPPO 170, the 

### 13 semi-field studies - I will elaborate on that later on - the 

### 14 semi-field feeding studies that have been mentioned here before 

### 15 in my opinion are not really appropriate as Tier II. 

### 16 **DR. DANIEL SCHLENK:**Dr. Hunt? 

### 17 **DR. GREG HUNT:**I'm not sure I understand you, 

### 18 Jens. You're saying that in Tier II, you don't think 

### 19 supplementary feeding studies have any place? 

### 20 **MR. JENS PISTORIUS:**Thank you, I will 

### 21 elaborate on this. I think three different types of studies 

### 22 have been mentioned. Please correct me if I'm wrong. First, 

### 23 the Oomen method. The second one is free-flying colony study, 

### 24 so it's a field, but still with supplementary feeding. Then you 

### 25 can do sugar feeding or pollen package feeding. I think this is 

### 26 not appropriate for Tier II tests.

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1 I think the most appropriate is to have a crop 

### 2 that offers nectar and pollen to over spray bees. During 

### 3 flowering, you get contact and oral exposure. With the colony 

### 4 that you feed either sugar syrup or pollen, you'll definitely 

### 5 have bees that go somewhere else and you will not be able to 

### 6 quantify the exposure. 

### 7 You will have a portion of bees even if you 

### 8 can say, okay the sugar water has been consumed and pollen has 

### 9 been eaten to this extent would never be able to quantify what 

### 10 exact amount has been. And it's not a worst case exposure 

### 11 anymore, which is the aim of a Tier II test in my opinion. 

### 12 So, with those feeding methods, I'm not happy 

### 13 at all. Once exception - the Oomen test can be used, but it's 

### 14 regarded as intermediate between Tier I and Tier II. The big 

### 15 advantage, why I still like it a little bit, is that you can 

### 16 actually do such a dose response. The test method, as it is 

### 17 described in the literature in 1992 is not appropriate. It 

### 18 needs modifications. For instance, there is a dosing of one 

### 19 liter sugar solution, a unique dosing. And this is not 

### 20 appropriate. It should be dosing over a longer time with syrup, 

### 21 at least, whatever at least nine or ten days that they get 

### 22 everyday sugar that you actually - and the big advantage is you 

### 23 can find some dose response via this. 

### 24 With the other ones, I think you are really 

### 25 neglecting something. If you do a feeding method, you cannot 

### 26 quantify exposure and there is a lot of worst case exposure.

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1 **DR. DANIEL SCHLENK:**Dr. Hunt? 

### 2 **DR. GREG HUNT:**I guess that clarifies it 

### 3 because you're not talking about tunnel experiments. In a 

### 4 tunnel experiment, you could quantify exposure. We just set one 

### 5 up last week. If you think that the exposure will be through 

### 6 pollen, then you establish colonies with depleted pollen and you 

### 7 feed them paddies and you measure what they eat. 

### 8 **DR. DANIEL SCHLENK:**Okay. Mr. Pistorius? 

### 9 **MR. JENS PISTORIUS:**When I get a study, I'm 

### 10 happy when I get a study that gives me a lot of information and 

### 11 not only partial information. What I would suggest to use EPPO 

### 12 170 and OECD 75 instead of such feeding studies. The reason for 

### 13 this is that you have a combined exposure. You will not be able 

### 14 to get a contact exposure of the bees. 

### 15 With a Tier I, we look at what is the toxicity 

### 16 for oral exposure, what is the oral toxicity, contact toxicity, 

### 17 but by making a spray application on the flowering crop where 

### 18 the bees are actively foraging. This is the only possibility 

### 19 where you have the option to get both effects of oral and of 

### 20 contact exposure in one goal. With this feeding study, if you 

### 21 identified a risk only for oral exposure, okay. But trying to 

### 22 get that we want to have the worst case situation, which I 

### 23 believe should be still the Tier II test, then I think those 

### 24 methods of feeding are not really giving us the worst case. 

### 25 **DR. DANIEL SCHLENK:**Dr. Tarpy? 

### 26 **DR. DAVID TARPY:**This is Dave Tarpy. So

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1 then, from what I'm hearing then is that that's what you're 

### 2 arguing that a full-field rather than a semi-field assessment is 

### 3 best, which I'm not disagreeing with, but what we're talking 

### 4 about here are Tier II semi-field tests. So, they're not 

### 5 supposed to recapitulate all of nature, you're supposed to be an 

### 6 intermediate right? So, while I agree that by forces feeding or 

### 7 by supplemental feeding to kinda of impose exposure may not 

### 8 always mimic the proper exposure routes. I think the question 

### 9 here is do they still have some utility. So, I don't think the 

### 10 question is are they mimicking realistic means of exposure, but 

### 11 do they have some sort of utility in effecting final colony 

### 12 outcome or final colony phenotype. I would probably say that 

### 13 they do, but I agree that they do not perfectly mimic how they 

### 14 would be exposed in the full field, right? But this is not full 

### 15 field, this is a semi-field Tier II level. 

### 16 **DR. DANIEL SCHLENK:**Mr. Pistorius? 

### 17 **MR. JENS PISTORIUS:**I just want you to 

### 18 clarify for me a Tier II is always a semi-field intolerance, not 

### 19 free flying. Free flying in nature, whatever, that for me is a 

### 20 Tier III test. 

### 21 **DR. DAVID TARPY:**This is David Tarpy again. 

### 22 While in the White Paper, they articulate and discuss the two 

### 23 types of Tier II studies. One type is in tunnels, and that's 

### 24 correct. The other is not confined spatially, right? So, 

### 25 that's what I was referring to in that. 

### 26 **MR. JENS PISTORIUS:**You are right, and this

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1 is exactly what I propose to be changed or what I personally 

### 2 feel is not appropriate to consider free-flying colonies as Tier 

### 3 II tests. 

### 4 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 5 **DR. JEFF PETTIS:**Along this line then, if and 

### 6 again I think in the White Paper, they do talk about a possible 

### 7 Tier II with feeding meter, either protein or syrup. If we move 

### 8 that to a Tier III study, do you still think it's not valid to 

### 9 at least have some level of exposure where you reduce the stores 

### 10 in the colony prior to the starting of the test and you start to 

### 11 provide supplemental syrup and/or protein over time? They are 

### 12 free flying and I agree -- I know one of the comments is going 

### 13 to be reduction of incoming pollen is not going to be absolute. 

### 14 We get dearth periods here in the mid Atlantic of two or three 

### 15 months where there is virtually no incoming nectar. So, we do 

### 16 get a period when we can do these tests and not have much 

### 17 incoming outside resources. So, we are providing the resources, 

### 18 we've limited the amount at that start, and we do feel that 

### 19 we're getting exposure. I just think that by controlling the 

### 20 dose a bit, we are in some cases doing maybe a worse case, I 

### 21 don't know. 

### 22 **DR. DANIEL SCHLENK:**Mr. Pistorius? 

### 23 **MR. JENS PISTORIUS:**I think it is actually 

### 24 what you mentioned, you are right. I think for some cases it 

### 25 can actually be used in the field conditions. For instance, now 

### 26 let's assume that we have a very highly toxic substance and it's

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1 even systemic and you have very much concerns and you want to 

### 2 have this long-term duration really checked. 

### 3 I think as an additional method after you have 

### 4 had an exposure to the real crop - I mean for instance, let's 

### 5 have a look at systemic and you have whatever we have in 

### 6 Germany, a winter oilseed rape crop, which is attractive and 

### 7 offering nectar and pollen. Then you say, okay maybe the test 

### 8 crop didn't flower long enough and in real life, you maybe have 

### 9 different flowering crops and this is the situation which is 

### 10 very hard to cover when you conduct a field test. To time the 

### 11 flowering of different crops, I mean that is maybe reality, but 

### 12 if you want to design the study, that is not possible. 

### 13 So then, I would agree that you can say, okay 

### 14 at field realistic concentrations, you can add pollen paddies 

### 15 and you can also do sugar feeding to extend exposure. To have, 

### 16 you know, a long-term test and to mimic that the exposure is not 

### 17 like winter oilseed flowering taking place four weeks, you can 

### 18 mimic that the exposure would be continuous over five weeks to 

### 19 get a worst case experiment. So I think, this case, additional 

### 20 pollen and nectar feeding is appropriate, but I think it's 

### 21 different study design from how I understood -- maybe I 

### 22 understood it wrong from how I understood what was described. 

### 23 **DR. DANIEL SCHLENK:**Okay. Dr. Hunt? 

### 24 **DR. GREG HUNT:**I think I better understand 

### 25 what Dr. Pistorius was taking about. You consider Tier II 

### 26 tunnel experiments, and I agree that is probably the most

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1 practical way to go. You have the most control. It's going to 

### 2 depend on the crop and what you think is the route of exposure. 

### 3 Some crops, the bees might not forage on in the tunnel, and in 

### 4 some cases it may be appropriate to do a feeding experiment. 

### 5 **DR. DANIEL SCHLENK:**Okay. Mr. Pistorius, we 

### 6 need to get on with this. Go ahead. 

### 7 **MR. JENS PISTORIUS:**Just for crops which are 

### 8 not attractive to bees, it is a question of systemic seed 

### 9 treatments for instance. I mean, if it's not attractive to 

### 10 bees, then the question is would you actually have exposure via 

### 11 nectar and pollen, which would then, as I understood, would not 

### 12 be the case. Then if you look at other issues, for instance if 

### 13 you have some guttation issue or whatever, you can do that. 

### 14 Maybe then, you can do some additional feeding. That's okay, 

### 15 maybe. But if you have a target crop where you want your 

### 16 application on, at least what we do and this target crop is not 

### 17 attractive to bees and we want to test the impact of a spray 

### 18 application, we use a surrogate flowering crop, which offers 

### 19 nectar and pollen because we want to know what the pesticide 

### 20 does. If it is intended not to be used on or if it's intended 

### 21 to be used on a not flowering crop, which offers no nectar and 

### 22 pollen, then this is basically a risk mitigation measure. 

### 23 **DR. DANIEL SCHLENK:**Okay. Any other comments 

### 24 on this? It sounds like that tunnels everybody is good with, 

### 25 but there may be some issues relate to the semi-field. Is that 

### 26 what I'm hearing? Yes? Mr. Pistorius?

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1 **MR. JENS PISTORIUS:**May I? Sorry. It's not 

### 2 tunnel or semi-field for me. For me, a semi-field is a tunnel 

### 3 study. It is the question, are they free flying and is 

### 4 additional syrup fit as a main exposure. 

### 5 **DR. DANIEL SCHLENK:**So, let's just be sure we 

### 6 document that very well in the written comments. Dr. Tarpy, 

### 7 you're responsible for that, I guess. Any other comments on 

### 8 this particular issue? Okay. Let me go to the agency then. 

### 9 Any specific questions of clarification? I will give you a 

### 10 moment or two. 

### 11 **MR. JOSEPH DECANT:**This is Joe DeCant, EFED. 

### 12 I think that - so question A refers to the tunnel design and 

### 13 specifically the tunnel design in question. B is going to touch 

### 14 on the feeding design that Mr. Pistorius was talking about. I 

### 15 think that maybe for clarification - so what I heard were the 

### 16 strengths and the weaknesses of the tunnel design sort of at the 

### 17 first part of the conversation and the discussion. I think what 

### 18 would be helpful is if the question is about where the feeding 

### 19 design fits into the tiered approach, if the Tier II is 

### 20 semi-field and we're talking about tunnel studies, then is the 

### 21 feeding design, does it sit in between Tier I and Tier II or 

### 22 does it sit in between the tunnel and the Tier III full field? 

### 23 Maybe just some clarification on where that might sit and 

### 24 getting to the utility of it in subpart (d). 

### 25 **DR. DANIEL SCHLENK:**Do you want and answer 

### 26 for that now? Yes? Can we answer that now? Dr. Tarpy?

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1 **DR. DAVID TARPY:**This is David Tarpy. Since 

### 2 I'm lead, I guess I'm the one in the crosshairs on that. I 

### 3 don't know if there is a clear answer to that. I think in some 

### 4 ways, it depends on the experimental design. A lot of these 

### 5 boundaries are blurred because of the social organism that we're 

### 6 using here as a model for it. In some cases, I would say that 

### 7 it lies between the Tier I and the food house design. In 

### 8 others, I would say it would not be. So, I don't know if there 

### 9 is a definitive answer to that question, if I'm understanding it 

### 10 correctly. 

### 11 **DR. DANIEL SCHLENK:**Yes, Mr. Sappington? 

### 12 **MR. KEITH SAPPINGTON:**Keith Sappington. With 

### 13 regards to subpart (a) and as you draft the report, it would be 

### 14 very helpful to the agencies for any insights that you can 

### 15 provide on the likelihood of false negatives with tunnel 

### 16 studies, semi-field tunnel studies as well as false positives, 

### 17 especially if this may end up being sort of the only study being 

### 18 considered in Tier II. Thank you. 

### 19 **DR. DANIEL SCHLENK:**Okay. Yes, Dr. James? 

### 20 **DR. ROSALIND JAMES:**The tunnel studies I've 

### 21 done have been with solitary bees, and I assume these problems 

### 22 are exacerbated with honey bees in that say with bumble bees, 

### 23 you have to feed them inside a greenhouse even when they're used 

### 24 for greenhouse pollination. So, the idea of being able to treat 

### 25 the plant in the tunnel and then expect normal foraging, you're 

### 26 not really going to get normal foraging because you have to have

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1 a really big tunnel. Bees need a lot of plants. There is not 

### 2 very much nectar and pollen produced per flower. So, they 

### 3 really need many, many plants and they quickly deplete out a 

### 4 greenhouse. So your dose may be reduced in a tunnel test. 

### 5 **DR. DANIEL SCHLENK:**Okay. Mr. Pistorius? 

### 6 **MR. JENS PISTORIUS:**I agree, maybe a 

### 7 different issue with bumble bee colonies, although we have also 

### 8 done some studies with bumble bee colonies, for instance oilseed 

### 9 rape or Phacelia tanacetifolia, which also offer good nectar. 

### 10 At least for the time of flowering, you have good exposure and 

### 11 you were able to keep the colonies maintain - I mean of course, 

### 12 you have to have a certain amount of food inside a bee colony, 

### 13 just at least a few hundred grams of honey and a little bit of 

### 14 pollen. So if there is a weather change for a few days, they 

### 15 don't starve to death because that will of course, kill your 

### 16 study. 

### 17 On the other hand, I think that this is still 

### 18 appropriate. You don't have normal foraging behavior. I agree 

### 19 because the tunnels may be of limited or they are always of 

### 20 limited size, but you can keep them and you can document that by 

### 21 the income of nectar and pollen by when you do the brood 

### 22 assessment that actually pollen came in. When you do the 

### 23 assessments for instance on bee brood, you can actually see at 

### 24 least they were able to, they were consuming the pollen that 

### 25 they had before, then they stored new pollen and they were able 

### 26 to take care of the brood for a while.

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1 Then to clarify this again, I think this is a 

### 2 good way of exposing colonies, but then if you want an extended 

### 3 exposure over the duration of the flowering crop, you can do 

### 4 additional feeding. That's what I thought. And what you said 

### 5 with greenhouse application, that may be true that you have for 

### 6 instance for tomato pollination, that you will have bumble bees 

### 7 starving a little bit and that you have to feed something 

### 8 additionally. Maybe it's a different system. I don't know. 

### 9 What we do is we register the substance. We register a product 

### 10 and we have to test that this product is safe, not just special 

### 11 application. So, what we would go for is we make an application 

### 12 on a flowering crop that is attractive to bumble bees, like 

### 13 Phacelia and would use these results to interpret the potential 

### 14 effects of the product on bumble bee colonies. Then what you 

### 15 can do to extend this again, you do some feeding. 

### 16 **DR. DANIEL SCHLENK:**Dr. James? 

### 17 **DR. ROSALIND JAMES:**So, it would help if you 

### 18 used something like the Celia that is a very high nectar and 

### 19 pollen producing plant instead of the crop. But still, if you 

### 20 have to start feeding at the end, you're effectively diluting 

### 21 your dose. Well, you're reducing the dose if you're adding - if 

### 22 you have to feed supplemental nectar and pollen, they're not 

### 23 collecting it from a crop that was treated. 

### 24 **MR. JENS PISTORIUS:**This is a very good 

### 25 point, thank you. When I mean additional feeding is possible 

### 26 for extending the exposure, of course I mean that you have to

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1 use spiked food with realistic field concentrations. Because I 

### 2 agree if you then feed uncontaminated sugar or pollen paddies, 

### 3 that is again a reduction of the exposure. This is basically 

### 4 reducing the certainty of the test. Then you get uncertainty 

### 5 and we don't know how to make the interpretation. 

### 6 **DR. DANIEL SCHLENK:**Okay. Mr. Sappington? 

### 7 **MR. KEITH SAPPINGTON:**Just one clarification. 

### 8 Mr. Pistorius, when you talk about potentially extending the 

### 9 study with feeding, is that being considered in the context 

### 10 within a tunnel or a separate study like a feeding study where 

### 11 they have free-flying bees. 

### 12 **MR. JENS PISTORIUS:**It would be considered 

### 13 for after the exposure to the flowering crop, so it's not an 

### 14 extra study. It is with the same colony. After the exposure 

### 15 period or after the forage in the tunnel is finished, the 

### 16 flowers are all gone basically, then what you could do is you 

### 17 add, have to think about opening up that tents so they are free 

### 18 flying or maybe you can close, but then you add basically sugar 

### 19 solution, spiked sugar solution and spiked pollen to extend this 

### 20 period over the blooming period if you have such a concern that 

### 21 it is in real life maybe more covering the realistic situation. 

### 22 23 **DR. DANIEL SCHLENK:**Okay. Any other 

### 24 questions of clarification from the Agency? Okay. Dr. Tarpy, 

### 25 you want to - oh sorry, first we need to read in the next 

### 26 question please.

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1 **MR. JOSEPH DECANT:**Joe Decant, question 11, 

### 2 subpart (b) - please comment on any other types of colony-level 

### 3 studies that should be considered as part of Tier II. 

### 4 **DR. DANIEL SCHLENK:**Dr. Tarpy? 

### 5 **DR. DAVID TARPY:**This is David Tarpy. So, 

### 6 the discussants talked about three kind of general alternatives 

### 7 that may be used, not really, well in Tier II as far as inside 

### 8 hoop houses or in cage studies. One is to utilize the power of 

### 9 observation hives, which are glass-walled colonies, glass-walled 

### 10 hives that enable observations of individual nest mate 

### 11 behaviors. So, comparable to what we were discussing in 

### 12 question number 8 this morning about how nest mate interactions 

### 13 and individual behaviors has relevance to colony level 

### 14 phenotype. 

### 15 Recapitulating that in a type II or a Tier II 

### 16 environment that could possibly augment those same types of 

### 17 things done in Tier I level may very well be helpful. Again, I 

### 18 think that the exact behaviors and the manner by which those 

### 19 behaviors are recorded and analyzed is a data gap, but I think 

### 20 that it is something that should be considered in these colony 

### 21 level studies, part of Tier II. 

### 22 Secondly as was eluded to in previous 

### 23 questions about the blurring of Tier I and Tier II when it comes 

### 24 to the colony reproductives. So, queens and drones are both 

### 25 very, very important. Unlike workers, which may be more 

### 26 amenable to small groups in vitro, it's very, very difficult to

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1 do that with queens and drones. So, Tier II studies ought to 

### 2 consider and incorporate means by which phenotypic effects on 

### 3 individual queens and drones can be measured, because those are 

### 4 in some ways a consequence of colony level phenotype. So 

### 5 measuring the reproductive potential of queens, the reproductive 

### 6 potential of drones is something that could also be folded into 

### 7 this level of assessment. 

### 8 Then finally, there was discussion earlier 

### 9 this morning about possible Tier II type studies that are done 

### 10 in vitro, right? So, having small colonies with functional 

### 11 queens, workers of all ages and brood that they're actively 

### 12 raising, but done in incubators and not actually in hoop houses 

### 13 or done outside, but in more controlled environments. These 

### 14 could probably be done for a period of days, may be weeks 

### 15 depending on the size and other factors. 

### 16 So these are, what I would consider Tier II 

### 17 studies because they're involving whole-level colony phenotype, 

### 18 but they're actually done in vitro. So again, it's this 

### 19 blurring of these different tiers when it comes to looking at 

### 20 super organisms. 

### 21 **DR. DANIEL SCHLENK:**Okay. Thank you. Dr. 

### 22 Ostiguy? 

### 23 **DR. NANCY OSTIGUY:**I concur. 

### 24 **DR. DANIEL SCHLENK:**And Dr. Pettis? 

### 25 **DR. JEFF PETTIS:**I concur with the comments 

### 26 of Dave. I will just add a bit about some of these potential in

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1 vitro kind of cage trials, it would more mimic a whole colony. 

### 2 We actually did some of those where we took a small - 100 

### 3 workers and a queen and introduced a known number of eggs to the 

### 4 colony and asked them to rear them and looked at the success 

### 5 rate of that small unit to rear that brood. It was pretty 

### 6 telling. I mean, you can get results with a fairly simple set 

### 7 up. They're not refined. They're not, in all cases - but I 

### 8 think that is something that moves between straight Tier I and 

### 9 Tier II where we begin to capture the super organism. 

### 10 **DR. DANIEL SCHLENK:**Any other panel members? 

### 11 Yes, Dr. Hunt? 

### 12 **DR. GREG HUNT:**Obviously, we're throwing out 

### 13 a lot of ideas on possible assays, and only a few of these will 

### 14 probably end up being the ones considered until there is further 

### 15 evidence of their effectiveness. But one possibility for 

### 16 another one is to - since there is a general feeling among 

### 17 beekeepers and bee researchers that perhaps we're seeing more 

### 18 problems with queens lately. There are indications that certain 

### 19 compounds like the miticides that beekeepers use in their hives, 

### 20 the queens are more sensitive to. 

### 21 So, one study that can be done in two weeks' 

### 22 time is to rear queens in a somewhat contaminated hive. This 

### 23 can be done very rapidly. The question is what is the best 

### 24 method to contaminate the colony, and I see Dr. Pistorius is 

### 25 raising his hand, so he has an idea on this. 

### 26 **DR. DANIEL SCHLENK:**Mr. Pistorius?

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1 **MR. JENS PISTORIUS:**I do have an idea. What 

### 2 you could do is make a shook swarm, which is basic for the 

### 3 non-beekeeper that you put the bees without the queen in a cage, 

### 4 in a box and then you put this box into a tent, and you take 

### 5 very, very low food reserves or if it's good weather, you can 

### 6 even do no food reserves, wait one or two days and then you put 

### 7 the queen frame in. Before this or one day before you put the 

### 8 queen frame in, you make your application so you ensure that the 

### 9 bees have collected nectar and pollen already. 

### 10 So, you know that that pollen and nectar 

### 11 inside is already contaminated and that they will be forced 

### 12 during the rearing of the queen to forage on this contaminated 

### 13 crop. Then, that is actually quite a short process until the 

### 14 queen cell is capped. You will definitely have the flowering 

### 15 period still, and then you can take them out and you can observe 

### 16 them, because then the queen is not fed anymore and then you can 

### 17 do additional experiments to the queen if you would like, but 

### 18 that's actually quite short and quite efficient test I would 

### 19 say. I don't know. 

### 20 **DR. DANIEL SCHLENK:**Dr. Hunt? 

### 21 **DR. GREG HUNT:**I agree. 

### 22 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 23 **DR. JEFF PETTIS:**Jeff Pettis. And I all 

### 24 ready want to modify the method. No. You don't like feeding 

### 25 nectar and spiked pollen, but I think this might be a perfect - 

### 26 rather than have them forage on the crop, if you know something

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1 about the residue in the nectar and pollen, why not just 

### 2 simulate that in a shook swarm, and just keep them completely 

### 3 artificial in little cages by themselves and introduce the 

### 4 queen. I agree, it's a great idea that asking a group of 

### 5 queenless bees to rear queen cells should be a great bioassay. 

### 6 **DR. DANIEL SCHLENK:**Mr. Pistorius? 

### 7 **MR. JENS PISTORIUS:**And I agree with Dr. 

### 8 Pettis. In this case, I would actually think if you know the 

### 9 concentrations that you have to spike nectar and pollen, that's 

### 10 a great idea then in a confined tunnel condition. 

### 11 **DR. DANIEL SCHLENK:**Dr. James? 

### 12 **DR. ROSALIND JAMES:**I don't know if this 

### 13 would really be considered a Tier II test. Throughout this, 

### 14 I've been wondering could you require at the Tier II level that 

### 15 a field experiment be conducted without bees in which then you 

### 16 measure actual residues in the pollen and nectar from different 

### 17 types of applications. That will give you some of these unknown 

### 18 answers, well how much then do we feed to bee. 

### 19 **DR. DANIEL SCHLENK:**Okay. Anyone else? 

### 20 Okay. Let me go to the agency. Any questions of clarification? 

### 21 We're good? Okay. Let's move onto letter C. 

### 22 **MR. JOSEPH DECANT:**Joe DeCant, question 11, 

### 23 subpart (c) - please comment on the most important endpoints 

### 24 that should be measured in the Tier II studies, such as adult 

### 25 forage bee mortality, brood development, queen fecundity, 

### 26 overall colony strength, that are linked to assessment endpoints

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1 and their associated protection goals. 

### 2 **DR. DANIEL SCHLENK:**Dr. Tarpy? 

### 3 **DR. DAVID TARPY:**This is Dave Tarpy. We 

### 4 discussed this briefly and largely agreed that those that were 

### 5 outlined in the White Paper were very adequate at looking at 

### 6 amount of brood, the overall colony population and strength, all 

### 7 of those measures of colony phenotype are very appropriate, and 

### 8 are a good way to quantify colony level effects. I should note 

### 9 I should note that by and large, all of those 

### 10 different measurements are highly correlated in and of 

### 11 themselves. So, quantifying a composite colony phenotype in 

### 12 these ways can be done statistically, principle component 

### 13 analyses, using different loading variables into that is a way 

### 14 to - is a means of comparison and is quite effective in doing 

### 15 that. 

### 16 To touch on the points that were just raised 

### 17 about looking at queen and don't forget drones - right, drones 

### 18 as well as queens - queen and drone viability and their 

### 19 individual phenotypes is another outcome and endpoint that 

### 20 should be measured in these types of Tier II studies as well. 

### 21 A third and final major endpoint is to sample 

### 22 and process returning foragers and to actually see what the 

### 23 foraging behavior is, quantifying their exposure rates, 

### 24 quantifying what their foraging on, and looking at the returning 

### 25 foraging force in these Tier II studies to really quantify their 

### 26 potential exposure routes.

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1 **DR. DANIEL SCHLENK:**Okay. Dr. Ostiguy? 

### 2 **DR. NANCY OSTIGUY:**I think this is also a 

### 3 point where some in-hive samples could be taken to potentially 

### 4 supplement. We had information on the contamination at the 

### 5 pollen and nectar to then look at the corresponding 

### 6 concentrations of the pesticides within honey and bee bread. 

### 7 **DR. DANIEL SCHLENK:**And Dr. Pettis? 

### 8 **DR. JEFF PETTIS:**To extend what Nancy said 

### 9 just a bit further, at the end of the exposure period within a 

### 10 tunnel in addition to looking at bee bread an pollen, you can 

### 11 take newly emerged adult bees that have been fed and processed 

### 12 during that exposure period and assay them directly, or then put 

### 13 them in cages and do longevity studies. A fairly easy thing to 

### 14 do, at the end of the study, doesn't add much to the cost. 

### 15 **DR. DANIEL SCHLENK:**Dr. James? 

### 16 **DR. ROSALIND JAMES:**Question for Dr. Tarpy. 

### 17 So, you said sample then process returning foragers to see what 

### 18 they're foraging on, but in a tunnel study, we pretty much know 

### 19 what they're foraging on, no? 

### 20 **DR. DAVID TARPY:**Sorry, this is Dave Tarpy. 

### 21 One can quantify the nectar loads of individual bees by 

### 22 capturing them at entrances, anesthetizing them with carbon 

### 23 dioxide, then squeezing their crop contents into capillary 

### 24 tubes, then very accurately measuring the nectar loads of each 

### 25 forager and then sampling many foragers to get an idea of the 

### 26 variation in how many are foraging on nectar versus pollen, what

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1 the average nectar loads are. You can even look at sugar 

### 2 content and other things. This is again, very well documented 

### 3 in the literature. That's what I meant is actually to sample 

### 4 the incoming foragers to see kind of more quantitatively of what 

### 5 their exposure rates are. 

### 6 **DR. DANIEL SCHLENK:**Okay. Any other Panel 

### 7 members? Mr. Pistorius? 

### 8 **MR. JENS PISTORIUS:**Real short - for me, this 

### 9 list is missing behavior at the hive entrance and behavior of 

### 10 foraging bees on the crop. 

### 11 **DR. DANIEL SCHLENK:**Okay. Make sure that we 

### 12 get that on the list. Okay. It seems pretty straight forward, 

### 13 but I'll ask if you guys have any questions of clarification? 

### 14 Yeah, Mr. Sappington? 

### 15 **MR. KEITH SAPPINGTON:**Keith Sappington. 

### 16 Quick question regarding the nectar loads in measuring those. 

### 17 Would that include measurement of residues in the nectar as well 

### 18 from the tunnel studies? 

### 19 **DR. DAVID TARPY:**This is David Tarpy. I 

### 20 assume that that could be done as part of that, yeah. 

### 21 **DR. DANIEL SCHLENK:**Okay. Let's go ahead and 

### 22 move onto letter D. 

### 23 **MR. JOSEPH DECANT:**Joe DeCant, question 11, 

### 24 subpart (d) - section 4.2.2 of the White Paper discusses a 

### 25 full-field feeding design. This methodology is discussed under 

### 26 Tier II assessments since the colony is relatively confined to

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1 foraging on either spiked sucrose solutions or spikes pollen. 

### 2 The intent of this methodology is to ensure that colonies are 

### 3 exposed to known residue levels over longer durations than the 

### 4 semi-field tunnel study designs. A limitation to the study is 

### 5 that bees may simply store spiked food rather than consume it 

### 6 and that the reliance on a single source of food may introduce 

### 7 confounding effects, such as nutritional deficits into the 

### 8 study. Please comment on the environmental realism and utility 

### 9 of full-field feeding studies as a line of evidence in 

### 10 characterizing risk to honey bee colonies. 

### 11 **DR. DANIEL SCHLENK:**Okay. Dr. Tarpy? 

### 12 **DR. DAVID TARPY:**So, I feel like we've all 

### 13 ready kind of discussed this quite a bit. So, I don't know if 

### 14 my comments here are really going to be all the relevant, but I 

### 15 guess there is some disagreement on the panel here as to whether 

### 16 is kind of zero utility, some utility or a lot of utility. I 

### 17 guess what it comes down to is that a well-designed full-field 

### 18 Tier II study, I think, would provide information that would not 

### 19 be gleaned at a Tier I level. Again, it may not realistically 

### 20 mimic a full ecological type of realism, but I believe it would 

### 21 have some - this type of design would have some utility. 

### 22 Again, honey bee colonies, I would think, 

### 23 compared to tunnel studies where there are some limitations just 

### 24 on their behavior being amenable to being in small enclosures 

### 25 that they would be more amenable to these full-field types of 

### 26 designs. I think that by being in enclosures, one of the

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1 limitations that we discussed earlier in this question is the 

### 2 shortened duration where as a full-field design would facilitate 

### 3 longer measurement durations of colonies and for chronic levels 

### 4 of exposure that could be very beneficial, especially looking at 

### 5 colony population and turnover where brood raised under a 

### 6 particular exposure could then be measured as adults. 

### 7 So, extending it beyond 28 days, which I 

### 8 believe the White Paper was mentioning, spending it beyond that 

### 9 to try and quantify some of those phenotypes of individual bees 

### 10 that were raised under those particular environmental 

### 11 conditions. 

### 12 **DR. DANIEL SCHLENK:**Okay. Dr. Ostiguy? 

### 13 **DR. NANCY OSTIGUY:**One possible way to 

### 14 address the issue about whether or not the bees store the 

### 15 contaminated material and not use it to raise current brood 

### 16 would be to actually sample the colony to require some 

### 17 indication of what was in the colony in the first place in terms 

### 18 of brood food, honey, et cetera, and then have a measurement 

### 19 that told you how much of what was used would possibly have been 

### 20 from the contaminated material. 

### 21 **DR. DANIEL SCHLENK:**And, Dr. Pettis? 

### 22 **DR. JEFF PETTIS:**Recognizing the concerns of 

### 23 my esteemed colleague, Mr. Pistorius, I actually do like these 

### 24 studies. If we recognize it, they may not represent the contact 

### 25 element. I think given that we reduce the stores at the 

### 26 beginning of the study and then spike them with relative rates

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1 in nectar and pollen that they do have some value. Again, 

### 2 running them at a time of year when there is less incoming 

### 3 resources, so you have some control over the consumption rate. 

### 4 We have a distinct spring and fall pollen and nectar flow. So, 

### 5 there are times in the year when they can be done. 

### 6 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 7 **DR. MAY BERENBAUM:**I think just as integral 

### 8 to the inclination of bees to forage broadly geographically is 

### 9 their tendency to glean. One missing worry here - there is a 

### 10 concern about nutritional deficits and storage of compounds, is 

### 11 that force feeding eliminates that behavior. It's an incredibly 

### 12 sophisticated system of communication, decision making, force 

### 13 feeding does not capture that flexibility in bee behavior. So, 

### 14 yes, these force feeding studies can be informative if and only 

### 15 if there is an understanding of the natural disposition of bees 

### 16 to learn a verse of stimuli and avoid them in the future and to 

### 17 sample and detect and possibly chose not to feed. My experience 

### 18 is mostly with caterpillars and even caterpillars, when given a 

### 19 choice, can avoid toxic food. It's much more difficult to 

### 20 interpret a no choice test than a choice test in terms of its 

### 21 ecological reality. 

### 22 **DR. DANIEL SCHLENK:**Thanks. Dr. Pettis? 

### 23 **DR. JEFF PETTIS:**Good point, May. I guess in 

### 24 some of the studies that I've observed or been a part of, you 

### 25 look at consumption rate by the colonies and often they are 

### 26 equal, so you can at least - you know, again, they're not given

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1 a choice. You have a spiked syrup and non-spiked. At least 

### 2 there is no deterrent to feeding or obvious deterrent. 

### 3 **DR. DANIEL SCHLENK:**Mr. Pistorius, you have 

### 4 something to add on this one? 

### 5 **MR. JENS PISTORIUS:**I will add a little bit. 

### 6 Just from feeding studies, I think we have less of, for 

### 7 instance, repellency effect with feeding sugar solutions. So, 

### 8 you can actually - and maybe this is, you know, for some 

### 9 specific questions, but it should not be the only one for your 

### 10 lines of evidence if you have a good OECD and EPPO study. You 

### 11 can have such studies as additional information, but for 

### 12 instance, we know from all the laboratory toxicity tests that 

### 13 the bees will feed on this. Even with pyrethroids, which have 

### 14 repellency effect when you spray them on flowering crops, if you 

### 15 give a sugar solution to the colony, they will just consume it. 

### 16 They'll take whatever is there. 

### 17 **DR. DANIEL SCHLENK:**I hear Mary Poppins in 

### 18 there somewhere right? Any other comments on section D here? 

### 19 Yes, Dr. Hunt? 

### 20 **DR. GREG HUNT:**Well, I think that when you do 

### 21 the experiment this way in the field and feeding, you don't know 

### 22 what the dose is. You don't really know what the dose is, but 

### 23 possibly it could be useful for, as Dr. Tarpy said, for looking 

### 24 at that cohort of bees that was raised during the time you're 

### 25 feeding and they're receiving some unknown dose, field relevant 

### 26 dose, and see if there're effects.

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1 **DR. DANIEL SCHLENK:**Okay. Any other comments 

### 2 on letter D. Okay. Let me go to the Agency. Mr. DeCant, do 

### 3 you have any questions of clarification? 

### 4 **MR. JOSEPH DECANT:**Joe DeCant, EFED. I have 

### 5 one question of clarification. If we're talking about this 

### 6 field feeding design and we're talking about the cohort that was 

### 7 raised under those conditions and we talked about the ability of 

### 8 the bees to store pollen and nectar, to what extent does the 

### 9 Panel agree about just that cohort of bees or the actual 

### 10 duration of the study? I guess there's a question of the 

### 11 duration of the study when bees can store the food and consume 

### 12 it later. 

### 13 **DR. DANIEL SCHLENK:**Dr. Tarpy, you want to 

### 14 lead off on that? 

### 15 **DR. DAVID TARPY:**Is the question about what 

### 16 the duration should be? Or which cohort you're asking? 

### 17 **MR. JOSEPH DECANT:**Joe DeCant. I think it 

### 18 would be about which cohort to look at, which also relates to 

### 19 the duration of the study. And I say that because, you know, 

### 20 we're talking about measurement endpoints and I think - so the 

### 21 White Paper discusses about minimizing the number of times that 

### 22 we have to open up the hive to collect our measurement 

### 23 endpoints. So, I think if the Panel has any insight about what 

### 24 cohort to target, maybe a timing of the study if we're talking 

### 25 about the dearth period, if that's in the middle of the summer 

### 26 time and there is a honey flow in the spring and the fall, you

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1 know the duration of the study - and I guess ultimately this 

### 2 gets back to maybe a clarification about under what conditions 

### 3 would this type of study be most useful. If we're talking about 

### 4 longer duration, the measurement endpoints, when to collect 

### 5 those measurement endpoints and the cohort to target as you 

### 6 mentioned about, Dr. Tarpy. 

### 7 **DR. DAVID TARPY:**Okay. This is David Tarpy. 

### 8 Others feel free to chime in here, but I think that there are 

### 9 many cohorts that would be informative to this type of design. 

### 10 One would be those active foragers and nurse bees. So, say 

### 11 30-day-old bees and 10-day-old bees, during the time of 

### 12 exposure, during the time of feeding, and then several weeks 

### 13 later, emerging, marking and following of the cohort of bees 

### 14 that were larvae during the time of that exposure as well. 

### 15 So, minimizing the number of times going into 

### 16 the colony, I don't see that as a limitation or a problem. I 

### 17 just see that all of the different cohorts would be very 

### 18 informative in their different ways. So, spending the period of 

### 19 time - and plus you could go on for a very long period of time 

### 20 looking at then subsequent cohorts as well. 

### 21 So, there really is no definitive endpoint to 

### 22 this, it just comes down to tracking these things over time. I 

### 23 don't know if there is one particular time period that makes 

### 24 more sense than others. Certainly, just looking at one period 

### 25 of time is going to be limited in scope. 

### 26 **DR. DANIEL SCHLENK:**Dr. Pettis?

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1 **DR. JEFF PETTIS:**Kind of echo what David 

### 2 said. If you have a 20-day flowering period and you have 

### 3 colonies even in a tunnel study, you want to look at adults 

### 4 right during the flowering period, their behavior and affect on 

### 5 adult mortality. But this brood you want to target maybe the 

### 6 midpoint of the flowering period, 10 days in, as a start point. 

### 7 You need those nurse bees to process that food for about 10 days 

### 8 before they actually, before their glands are fully developed 

### 9 and they're feeding larvae. 

### 10 So, then you just keep moving the clock. 

### 11 We've been going out about 30 days beyond the midpoint of the 

### 12 flowering period to capture that. And of course, if you have 

### 13 opportunity to follow multiple cohorts, it's better. You may 

### 14 see during the peak of incoming nectar and pollen, you may see 

### 15 an effect. Then later, you may see that effect disappear. So, 

### 16 following multiple cohorts - I noticed in the White Paper that 

### 17 there was a concern about disruption to the colony. I think 

### 18 given very good beekeeping practices, that's not such an issue. 

### 19 You will see sometimes a spike in adult mortality if the person 

### 20 managing the hives is not careful, but I think the need to get 

### 21 the data probably overweighs and just hiring a good beekeeper is 

### 22 a way to avoid that. 

### 23 **DR. DANIEL SCHLENK:**Anybody else want to 

### 24 chime in on this one? Dr. Hunt? 

### 25 **DR. GREG HUNT:**Well, I think that some of 

### 26 this information can be gotten out of tunnel studies. I think

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1 there is some kind of consensus that tunnel studies are good, 

### 2 more controlled and should be the main focus of Tier II. If 

### 3 anybody disagrees, speak up now. 

### 4 **DR. DANIEL SCHLENK:**Yeah, in terms of writing 

### 5 this up, it sounds like it would be nice to present just exactly 

### 6 what you said. It seems like some people are for them, limited 

### 7 and the caveats as well as those, the reasons why they're not 

### 8 useful, I guess that would probably be a good way to put that in 

### 9 the minutes. Would that be helpful? Yeah? Dr. Pettis? 

### 10 **DR. JEFF PETTIS:**I guess along that line, do 

### 11 we feel as a group that the free-flying feeding studies are not 

### 12 - like you feel that they're not Tier II at all, they're really 

### 13 Tier III studies. Is there is any general consensus among the 

### 14 group about whether free-flying colony level studies with spiked 

### 15 stuff, is that more of a Tier III or more Tier II? Is there a 

### 16 feeling? 

### 17 **DR. DANIEL SCHLENK:**How many say it's a Tier 

### 18 III? Okay how many IIs? Look like we're split. So, sounds 

### 19 like it's a split. Looks pretty even actually. Pardon? Yes, 

### 20 you can clarify, Mr. Pistorius. 

### 21 **MR. JENS PISTORIUS:**With respect to what is 

### 22 the ultimate possibility for decision? So I guess it would 

### 23 actually be rather a Tier II. That was not my point. I think 

### 24 the study types are not the only ones that you can rely on and 

### 25 it was basically the study design that I was criticizing, but 

### 26 now in the discussions, I think we have elaborated some very few

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1 possibilities where actually a feeding study can also be useful 

### 2 or in addition to a really good OECD, EPPO, you can use this 

### 3 feeding to enhance. I hope that clarifies. 

### 4 **DR. DANIEL SCHLENK:**I'm sure we'll look 

### 5 forward to the written comments on this one. Dr. Hunt? 

### 6 **DR. GREG HUNT:**I want to change my vote. 

### 7 Yes, it's more of a Tier II, and clearly the experiment with 

### 8 raising queens would be Tier II because they're not free flying. 

### 9 **DR. DANIEL SCHLENK:**Okay. Noted. I guess 

### 10 that I think there is - are we II as well? 

### 11 **DR. JEFF PETTIS:**Last comment - I've all 

### 12 ready cast my vote. Last comment along these lines, I do feel 

### 13 that the EPPO guidelines and the OECD - there was a lot of 

### 14 thought and effort that goes into them. They are very valid 

### 15 things to follow. 

### 16 **DR. DANIEL SCHLENK:**Yeah, we can set this up 

### 17 in the minutes in terms of kind of a split decision and people 

### 18 can weigh in before it actually comes out at the end to get a 

### 19 final vote, whatever that is, whatever that means for you guys. 

### 20 Okay. Any other comments on this? Mr. DeCant, are you fine 

### 21 with what we've given you so far on that at this point? Okay. 

### 22 You want to read in letter E then? 

### 23 **MR. JOSEPH DECANT:**Joe DeCant, question 11, 

### 24 subpart (e) - as discussed in section 4.3.4 of the White Paper, 

### 25 it is important to consider the biological significance of a 

### 26 measured effect in addition to its statistical significance.

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1 Please comment on the nature and magnitude of effects that would 

### 2 be sufficient to conclude biologically significant effects on 

### 3 the colony and/or the need to transition to Tier III 

### 4 assessments. 

### 5 **DR. DANIEL SCHLENK:**Dr. Tarpy? 

### 6 **DR. DAVID TARPY:**This is David Tarpy. I 

### 7 think we agree 100 percent that reconciling statistical and 

### 8 biological significance can be a challenge. I would probably 

### 9 just want to leave it at that. This is an incredibly difficult 

### 10 question to answer with any sort of certainty. I mean, this is 

### 11 something that we all wrestle with of course, and it's even 

### 12 further blurred and confused by this kind of social nature of 

### 13 honey bees, and it just makes it even more difficult of a 

### 14 challenge because what can be very, very small, seemingly 

### 15 insignificant effects at the individual level can have profound 

### 16 ramifications at the colony level. 

### 17 Conversely, very large, seemingly striking 

### 18 effects between individuals can be balanced out and can be 

### 19 compensated for at the colony level. So, it is really, really 

### 20 difficult to try to reconcile this. So, I don't know if we're 

### 21 going to really be able to do justice to this particular 

### 22 question, but all of that said, we believe that power analyses 

### 23 that was asked for in the White Paper in multiple occasions at 

### 24 these different tiers feel like that's incredibly informative 

### 25 and should definitely be included in all of the results. 

### 26 This is especially true with the small sample

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1 sizes, small colony sample sizes because of the inherent 

### 2 variability across colonies in different contexts, different 

### 3 environments with larger variability, higher sample size is 

### 4 needed really to be able to elucidate statistically significant 

### 5 effects, which hopefully will provide inference to biologically 

### 6 meaningful effects. So, larger sample size is obviously a 

### 7 logistical limitation, but it's one that needs to be maximized 

### 8 with a sense of urgency, you know, at all possibility. 

### 9 There were some comments earlier talking about 

### 10 the Tier II studies and different measurements that would be 

### 11 taken. One that touches on this particular question about kind 

### 12 of teasing apart statistical versus biological relevant effects 

### 13 is doing ground truthing and looking at actual foraging behavior 

### 14 in the target crop or whatever the bees are actually foraging 

### 15 on. Those type of data, I think, would really help augment the 

### 16 interpretation of trying to tease apart statistical versus 

### 17 biological effects. 

### 18 Some other points here that I think we've 

### 19 already raised, so I won't go over those again. I think one 

### 20 last approach is trying to merge data sets or trying to 

### 21 consolidate data. One difficulty of having low sample sizes and 

### 22 lower power, if different data sets could be combined and have a 

### 23 meta analysis approach, that is a way to also reconcile this 

### 24 point that there may be small, even statistically nonsignificant 

### 25 effects that aren't detected within a single study, but by 

### 26 bolstering sample size by doing this similar study in different

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1 environments and different contexts, meta analyses can often 

### 2 pull out biologically relevant effects where each individual 

### 3 study might not have statistically meaningful effects. 

### 4 So, that again might be beyond the scope of 

### 5 this, but that is another avenue to consider in consolidating 

### 6 data. 

### 7 **DR. DANIEL SCHLENK:**Thanks. Dr. Ostiguy? 

### 8 **DR. NANCY OSTIGUY:**I'm going to make another 

### 9 plug for the analytical models because you can get two things 

### 10 from them. One data that may not show statistical significance 

### 11 or variables that may not show statistical significance may turn 

### 12 out to be incredibly biologically important. You may have 

### 13 somethings that the analytical model would tell you that you 

### 14 have statistical significance on that variable, but it doesn't 

### 15 really matter for the colony. 

### 16 **DR. DANIEL SCHLENK:**And Dr. Pettis? 

### 17 **DR. JEFF PETTIS:**Just very briefly, again we 

### 18 all feel better when we have high statistical power and we have 

### 19 good P values that lead to better publications and stuff like 

### 20 that. But, I don't think we can discount some of the biological 

### 21 effects that we see and I'll give you an example. 

### 22 So, you have a Tier I study, you're measuring 

### 23 mortality and other things, but you notice that the bees are 

### 24 excessively grooming, which could mean a response to pesticides. 

### 25 Then you notice the same response in a Tier II or a Tier III 

### 26 study. So, you see an effect, but it's statistically not valid.

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1 I think these biological effects that we see, we have to take 

### 2 them into consideration and try to figure out what they mean for 

### 3 the colony level. But we recognize that for many of these 

### 4 colony level studies, the ability to do adequate replication is 

### 5 hard to get there - economically it's hard to design a study 

### 6 well enough and have a enough replication to strong statistical 

### 7 power. 

### 8 **DR. DANIEL SCHLENK:**Okay. Any other panel 

### 9 members want to weigh in on this one? Dr. Hunt? 

### 10 **DR. GREG HUNT:**Just a comment. You're 

### 11 dealing with a bunch of academics here. So modeling and meta 

### 12 analyses are maybe good for discovery and for planning, but to 

### 13 echo what Jeff said, you know, I guess what we're really talking 

### 14 about is testing for risk analysis. So, there're two different 

### 15 things going on here. 

### 16 **DR. DANIEL SCHLENK:**Dr. Fefferman? 

### 17 **DR. NINA FEFFERMAN:**While modeling - so, I'm 

### 18 not going to speak so much to the meta analysis for statistics 

### 19 because honestly, I'm not a biostatistician, but while modeling 

### 20 has a huge role in discovery, it can also lead to 

### 21 recommendations for which things to test as measurements, and 

### 22 what those thresholds for the tests should be. So, I think it 

### 23 would be a mistake to dismiss as a purely academic concern that 

### 24 doesn't directly lead to applied recommendations. 

### 25 **Dr. DANIEL SCHLENK:**Any other comments? 

### 26 Okay. Go to the Agency, any questions of clarification? Keith?

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1 2 **MR. KEITH SAPPINGTON:**Keith Sappington. 

### 3 Yeah, this is a difficult question and it again, cuts across 

### 4 taxa, the biological and statistical significance and it would 

### 5 obviously differ depending on endpoints. 

### 6 Speaking from my own experience in reviewing 

### 7 half a dozen or so semi-field studies, I understand there is 

### 8 going to be a lot of gray area where best professional judgement 

### 9 is needed, but if there are bounds if we could put on, if 

### 10 there're areas where you say, look this is an obvious place 

### 11 where its biologically significant because we're dealing with 

### 12 endpoints like brood compensation index and brood termination 

### 13 rates as well as percent comb areas and various stages of brood. 

### 14 So to help us with that interpretation, I'm just speaking to 

### 15 the point of biological significance, that would be extremely 

### 16 helpful. 

### 17 **DR. DANIEL SCHLENK:**So if I may, it sounds 

### 18 like which biological endpoints have a more biological relevance 

### 19 than say a statistical relevance out of the ones that you're 

### 20 sort of measuring. Is that. 

### 21 **MR. KEITH SAPPINGTON:**Keith Sappington. 

### 22 Partly, yes, which endpoints maybe carry a little more 

### 23 biological weight, but even within an endpoint. For example, 

### 24 with the OECD studies 75, there are endpoints including brood 

### 25 termination rate and so forth, but there isn't, to my knowledge, 

### 26 a criterion of what's acceptable for that in the controls. So,

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1 there's interpretation around that. And then just understanding 

### 2 the biological significance, so if you do have a two-fold change 

### 3 regardless of what the stats can say, is that meaningful? And 

### 4 again, obviously that can get into the modeling, which we will 

### 5 be talking about, but any insights based on your experience with 

### 6 conducting these studies and knowledge of bee biology would be 

### 7 helpful. Thank you. 

### 8 **DR. DANIEL SCHLENK:**Dr. Tarpy, you want to 

### 9 come up with some guesses there? 

### 10 **DR. DAVID TARPY:**So would it be helpful to 

### 11 the Agency - sorry this is Dave Tarpy again - would it be 

### 12 helpful to perhaps rank or somehow qualify the list of different 

### 13 colony phenotypes on page 132 and 133 in the White Paper to say 

### 14 as biologically critical versus more environmentally 

### 15 temperamental or something like that where that would give you 

### 16 an idea of the gravity of the particular measurement, is that 

### 17 kind of what you're looking at as far as which are more 

### 18 biologically relevant versus less? 

### 19 **MR. KEITH SAPPINGTON:**Keith Sappington. Yes, 

### 20 if that can be done with some confidence, then that would be 

### 21 helpful, sort of a first tier versus maybe a second tier in 

### 22 terms of biological significance. But again, even within an 

### 23 endpoint, understanding daily mortality rates while I have a 

### 24 six-fold increase for three days, but then things return back to 

### 25 normal. I'm not asking for a specific answer on just that, but 

### 26 understanding even within an endpoint if there are bounds and

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1 very clear bounds on the magnitude that says yes, this is a 

### 2 problem. Like we had discussed earlier that perhaps a sustained 

### 3 mortality of 30 percent based on experience, it looks pretty 

### 4 major. So, if there are some bounds that we can put around the 

### 5 zone of uncertainty, that would be helpful, thank you. 

### 6 **DR. DANIEL SCHLENK:**Okay. We'll go with Dr. 

### 7 Berenbaum first and then Dr. Fefferman. 

### 8 **DR. MAY BERENBAUM:**Completely shooting from 

### 9 the hip, in as much as I have never done any studies. But just 

### 10 in terms of statistics and biological relevance, it would seem 

### 11 those parameters that relate to the queen who is not replicated 

### 12 in any colony should be given some latitude in terms of 

### 13 biological reality. Also, given the incredibly central role of 

### 14 the queen, you have on page 132 there are some measurements 

### 15 relating the presence of the same queen. I mean, within a 

### 16 single colony, you don't have that same statistical power that 

### 17 you do with even a small colony with 5 or 10,000 workers. So, I 

### 18 would say you should have some sort of - if something goes wrong 

### 19 with the queen and that is biologically relevant, let the bee 

### 20 biologists address that one. 

### 21 **DR. DANIEL SCHLENK:**Dr. Fefferman? 

### 22 **DR. NINA FEFFERMAN:**So I think we're 

### 23 conflating two different aspects into the same conversation, and 

### 24 one is - we're conflating two different elements. One is how to 

### 25 get from the statistical significance to the biological 

### 26 significance and the other is the criticality of the biological

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1 significance. 

### 2 I think Dr. Berenbaum's statements speak 

### 3 directly to how to get from the statistical to the biological, 

### 4 but it would be a mistake to try and intuit which aspects are 

### 5 critical biologically relevant aspects as opposed to transient 

### 6 biologically relevant aspects. Within the category of being 

### 7 biologically relevant just isn't off the cuff because we've got 

### 8 example after example from the last 50 to 100 years of research, 

### 9 of experts in the field with very good reasons and very sound 

### 10 science behind them going, I really think this is the important 

### 11 thing, later testing that in either modeling or as greater 

### 12 sensitivity of empirical techniques become available, realizing 

### 13 that the intuition is in fact really off. 

### 14 So, while we can do that, I think we have to 

### 15 separate the discussion from the statistical to the biologically 

### 16 relevant and then be very careful within the biological 

### 17 relevance of estimating relative criticality. 

### 18 **DR. DANIEL SCHLENK:**Okay. Dr. Ostiguy? 

### 19 **DR. NANCY OSTIGUY:**I'd like to add that in 

### 20 addition to queens, if something goes wrong with your drone 

### 21 population, you're in trouble. 

### 22 **DR. DANIEL SCHLENK:**Anybody want to add the 

### 23 other ones too? Dr. Pettis? 

### 24 **DR. JEFF PETTIS:**I'm glad to hear that female 

### 25 side speaking up for drones. That's encouraging. Not Greg, are 

### 26 we looking at here.

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1 **DR. DANIEL SCHLENK:**Except for Greg here. 

### 2 Greg's kind of. 

### 3 **DR. JEFF PETTIS:**Back to biological 

### 4 significance, even with the comment of Dr. Fefferman, I think 

### 5 one thing that I look at and Dave, I think we could take some 

### 6 stab at trying to prioritize to some degree, maybe in two 

### 7 categories. The one that jumps out at me always is just the 

### 8 level of brood production and brood survival. The reason that 

### 9 is so important is it incorporates so many other aspects of the 

### 10 hive activity. Incoming pollen and nectar, nurse bee activity, 

### 11 it really is a great measure. 

### 12 When we look - if we're measuring colony 

### 13 strength, we can measure adult population, which is harder to 

### 14 measure, but we can measure sealed brood production. It's real 

### 15 easy to measure very accurately as a very robust measure. So, 

### 16 if I had to look to one measure, it would be something about 

### 17 brood survival or brood production. But again, I agree that we 

### 18 can't over look everything that is out there. We have to 

### 19 consider that it is perhaps important. 

### 20 **DR. DANIEL SCHLENK:**Okay. Anybody else got 

### 21 another favorite measure? Yes, Mr. Pistorius? 

### 22 **MR. JENS PISTORIUS:**I was keeping back, 

### 23 because it is such a hard question and we ask ourselves a lot 

### 24 too. It depends on the level of protection that you want to 

### 25 achieve for your protection goals. If your major protection 

### 26 goal is the colony size, okay then you might have forager

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1 mortality event, which is not that relevant for colony survival 

### 2 and colony development over the long term. 

### 3 On the other hand, we are for instance, 

### 4 incident driven. We don't want incidents because that is part 

### 5 of my job, investigating the incidents. Don't need too much 

### 6 work. What we don't want, for instance, acute mortality events. 

### 7 So for us, it is a concern if we have clearly increased - it is 

### 8 again the question what level. But if we clearly increase 

### 9 mortality, there was a saying from a bird person from the 

### 10 Netherlands who said in the Netherlands, they don't want clearly 

### 11 visible dead birds. We want exactly the same for bees. We 

### 12 don't want clearly visible dead bees. That's why with the 

### 13 methodology of the bee trap we are a lot more protective 

### 14 compared to what the beekeeper actually realizes. 

### 15 If we see a clear increase in mortality, we 

### 16 even say okay, this is for us biologically relevant, even if the 

### 17 brood production is not yet affected. So for me personally, the 

### 18 most critical, in my personal top list, is acute mortality. 

### 19 Then I agree, it is not really below it on the same very 

### 20 important level is the brood production. Because depending on 

### 21 the substance, if you have an IGR, of course you would not get a 

### 22 clearly visible incident, but you will have no more brood. So, 

### 23 they are both basically on the same level. They are the most 

### 24 important ones in my opinion. 

### 25 Foraging behavior, okay this is also very 

### 26 important. For instance, if you would have a substance where

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1 you have a clear impact on and a long lasting impact on the wish 

### 2 of the bee to forage on that crop, that would of course affect 

### 3 one of your protection goals too, the quantity of hive products. 

### 4 So, I think this is also very important. We have to take them 

### 5 all into account. Ranking will be difficult, but those are I 

### 6 think the most relevant ones. 

### 7 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 8 **DR. MAY BERENBAUM:**I think maybe splitting 

### 9 hairs to say what's most or least relevant. But there's one 

### 10 profound difference in assessing nontarget impacts on bees than 

### 11 for birds or most other nontarget arthropods is that most 

### 12 people, even though the nontarget organisms can be beneficial. 

### 13 Most people don't make a living out of those beneficial 

### 14 organisms. 

### 15 So, in the case of nontarget impacts on bees, 

### 16 it's important economically not only that they are not dead, but 

### 17 that they are also functional and capable of both delivering the 

### 18 economic services for which they are being raised. So, it's a 

### 19 different context. I used the analogy earlier in a conversation 

### 20 in some ways and army, a dead soldier - no disrespect to the 

### 21 military - but a dead soldier at least is easier to deal with 

### 22 than one that's suffering mental of physical problems. 

### 23 So, in a way, we have these troops out there 

### 24 that are delivering pollination services. So, it's not just 

### 25 that they're numbers are reduced. If their behaviors are 

### 26 profoundly altered so that they are not delivering the

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1 ecological and economic services, for which they are being 

### 2 maintained. That's important in the context of risk assessment. 

### 3 4 **DR. DANIEL SCHLENK:**Dr. James? 

### 5 **DR. ROSALIND JAMES:**A point taken with the 

### 6 answer, you could go back to your protection goal and say what 

### 7 are your three protection goals and weigh them. So, what are 

### 8 they provision of pollination services, so what is going to be 

### 9 important for provision of pollination service. I will be hive 

### 10 activity but also be colony size and colony survival. That goes 

### 11 back to what you were saying about protecting the beekeeper 

### 12 also, and then similar for production of hive products will be 

### 13 the same factors, generally colony size and colony survival. 

### 14 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 15 **DR. JEFF PETTIS:**You beat me to it, Rosalind. 

### 16 But going back to those two protection goals, the first two 

### 17 protection goals, your point about loss of forager force in a 

### 18 pollination contract could be critical. So, a short impact on 

### 19 pollination or the pollinating force would be critical at that 

### 20 point, even though it may not have colony level effects, which 

### 21 would affect whatever protection goal number two. 

### 22 **DR. DANIEL SCHLENK:**Dr. James? 

### 23 **DR. ROSALIND JAMES:**But then I still have to 

### 24 add - the fecundity still keeps coming back. We had a case with 

### 25 a growth regulator and alfalfa leaf cutting bees. So a new 

### 26 growth regulator was approved for alfalfa, applications to

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1 alfalfa seed and after it was used in fields the first year we 

### 2 had the beekeepers as alfalfa leaf cutting beekeepers come to us 

### 3 and say we got very poor bee return, that is the number of bees 

### 4 that came back out of the field, from those fields that were 

### 5 treated with this product and be return was normal in the fields 

### 6 where the product wasn't used. The product had very low acute 

### 7 toxicity to adult bees. 

### 8 So then, some of the scientists in our group 

### 9 investigated it and well, if it's a growth regulator, it's not 

### 10 going to really affect adults, but maybe it'll affect fecundity. 

### 11 It turned out a closer investigation that the queens were 

### 12 laying inviable eggs, either that or the larvae exposed to the 

### 13 pollen provision was contaminated. We don't know which, but the 

### 14 eggs were not hatching or when they hatched, they died 

### 15 immediately. So, fecundity kind of comes into play always also. 

### 16 17 **DR. DANIEL SCHLENK:**Dr. Fefferman? 

### 18 **DR. NINA FEFFERMAN:**So, in some sense, I love 

### 19 all of these comments and I'm now almost stealing my own 

### 20 thunder, because you asked me to put examples into question 14. 

### 21 This was the example that I put in. We've been talking about 

### 22 rates of mortality as an issue and now we're talking about which 

### 23 things are most critical. It's a mistake to separate those two. 

### 24 Mathematically, if you had an analytic model for this, it is 

### 25 the ratio of worker death to worker recruitment. Whether that's 

### 26 from eclosion or egg laying, whatever is driving that, that

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1 ratio has to be greater than one in order for the colony to be 

### 2 successfully stable as a population demographic. And either 

### 3 side of that, either because the eclosion rates drop or because 

### 4 mortality goes up can drive that ratio less than one. 

### 5 That is why I'm really sort of cringing about 

### 6 identify these set of things as being critical because neither 

### 7 of those in isolation is necessarily critical to colony 

### 8 survival. It is critical - I like very much Dr. Pettis's idea 

### 9 if it is critical to the pollination services over a duration, 

### 10 but in terms of colony survival, neither one in isolation is 

### 11 itself critical. The ratio of the two is incredibly critical, 

### 12 and that is the sort of thing where just shooting from the hip 

### 13 on what seems to be critical gives you sort of the wrong insight 

### 14 into that. 

### 15 **DR. DANIEL SCHLENK:**So, let me just remind 

### 16 the Panel again that anything that you say, you need to get 

### 17 written comments to Dr. Tarpy on this particular issue, because 

### 18 he needs to put these together into some coherency. So, just a 

### 19 reminder there. Any other comments? Dr. Hunt? 

### 20 **DR. GREG HUNT:**Maybe I'll get the last word. 

### 21 We can't really rank these things as has been said, but anything 

### 22 the affects the reproductives would probably be at the top. 

### 23 Like if you get deformed queens, that's bad. 

### 24 **DR. DANIEL SCHLENK:**Okay. I think you're 

### 25 going to get a list, Keith, on a bunch of stuff. 

### 26 **MR. KEITH SAPPINGTON:**Thank you very much.

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1 **DR. DANIEL SCHLENK:**Any other questions, 

### 2 clarification questions from the Agency? Alright, let's take a 

### 3 break. Let's come back at a quarter to three. 

### 4 (WHEREUPON a recess was taken) 

### 5 We're ready to lead off with question 12, 

### 6 letter A. 

### 7 **MR. JOSEPH DECANT:**Joe DeCant, EFED, question 

### 8 12, subpart (a) - please comment on the strengths and 

### 9 limitations of full-field studies described in the white paper. 

### 10 **DR. DANIEL SCHLENK:**All right, Dr. Pettis, 

### 11 you're our lead discussant. 

### 12 **DR. JEFF PETTIS:**Thank you -- Dr. Pettis. 

### 13 Tier III studies are meant to be ultimate tests of real world 

### 14 exposure and potential effects. However, these studies are 

### 15 fraught with potential pitfalls with regard to the ability to 

### 16 carry out controlled experiments in the open environment and 

### 17 with adequate controls. 

### 18 When properly executed, Tier III studies are 

### 19 the best means to assess the risk and impacts of pesticides to 

### 20 Apis mellifera colonies and other managed colonies such as 

### 21 Bombus or Osmia. In the context of the regulatory framework, 

### 22 Tier III studies are considered highly refined from the White 

### 23 Paper. Studies meant to address specific concerns raised in 

### 24 lower tier studies, in that said, each study design should vary 

### 25 with regard to the endpoints that we measured to address these 

### 26 specific concerns that may be raised in Tier I or Tier II

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1 testing. 

### 2 So, I will point out some strengths and some 

### 3 weaknesses that the group felt were relative. So for strengths, 

### 4 the natural exposure that can be achieved by allowing bees to 

### 5 forage freely on the treated crop and then measuring endpoints 

### 6 allows for the testing of effects at the colony level that are 

### 7 unattainable at Tier I and Tier II level testing. The exposure 

### 8 achieved can accurately simulate the actual use of the 

### 9 pesticide, another strength. 

### 10 Tier III tests allow for colony-level testing 

### 11 of protection goals. So, it's the only aspect. Maybe a little 

### 12 bit in Tier II, but you can do colony-level test. If 

### 13 significant effects are noted at the colony level, the results 

### 14 can be interpreted with a high level of confidence as the colony 

### 15 has many buffering mechanisms such as reserve worker force, 

### 16 ability to compensate for lost or dead workers, things like 

### 17 that. An effect seen at the colony level is probably realistic. 

### 18 19 The use of surrogate crops such as Phacelia is 

### 20 possible the help increase foraging on the crop, or maybe the 

### 21 crop of interest, and thus insuring exposure. So those are some 

### 22 of the strengths. 

### 23 Some of weaknesses - study site selection is 

### 24 probably the single most critical factor in designing a decent 

### 25 study, competing vegetation can result in low or no exposure to 

### 26 the target crop and plot size can often be limited when

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1 designing these Tier III studies. 

### 2 Again, now plot size will vary with the 

### 3 attractiveness of the crop in question. So, if you have an 

### 4 unattractive crop, you may have to have a slightly larger plot 

### 5 size to ensure adequate exposure. 

### 6 As far as competition, bees will forage on the 

### 7 most attractive crop within the flight range, and this again can 

### 8 lead to inadequate exposure to the target crop. So, a knowledge 

### 9 of the surrounding area is critical and attempts to minimize 

### 10 competing vegetation is important. 

### 11 The need to treat the control plots to keep 

### 12 them healthy to some degree may confound the effects that you 

### 13 may be looking for. So, treating the control plots can be a 

### 14 major issue, but the lack of treating the control plot may make 

### 15 it non-comparable because of the health of the plants themselves 

### 16 may be compromised without treatment. 

### 17 Then lastly, positive control plots are 

### 18 normally not possible in an environment like this where you 

### 19 spray a highly toxic compound on a third set of plots to 

### 20 demonstrate an effect. Those are the things that I listed, the 

### 21 strengths and weaknesses. 

### 22 **DR. DANIEL SCHLENK:**Thanks. Dr. Ostiguy? 

### 23 **DR. NANCY OSTIGUY:**I concur. 

### 24 **DR. DANIEL SCHLENK:**Dr. Potter? 

### 25 **DR. THOMAS POTTER:**I have one comment. We 

### 26 may want to make this in several places, is the possible and

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1 potential confounding effect of the need to use other 

### 2 agrochemicals to have agronomic success of your crop may indeed 

### 3 make it difficult to interpret or evaluate the impacts of the 

### 4 active ingredient that's the focus of the investigation. 

### 5 **DR. DANIEL SCHLENK:**Okay. Dr. Tarpy? 

### 6 **DR. DAVID TARPY:**I have nothing further to 

### 7 add. 

### 8 **DR. DANIEL SCHLENK:**Other panel members? Dr. 

### 9 Ostiguy? 

### 10 **DR. NANCY OSTIGUY:**What I remember as talking 

### 11 about that, maybe it was with you Jens that we talked about how 

### 12 if you then applied the material that is applied to the control 

### 13 crop, you apply it to your treatment crop to attempt at least to 

### 14 remove that effect from the control crop. 

### 15 **DR. THOMAS POTTER:**You know, absolutely. You 

### 16 know, in terms of treating your control and hopefully keeping 

### 17 your materials limited, but even still those other products are 

### 18 stressors and may ultimately obscure what you're really try to 

### 19 measure is the effect of the ingredient you're working with. 

### 20 **DR. DANIEL SCHLENK:**That was Dr. Potter. Dr. 

### 21 James? 

### 22 **DR. ROSALIND JAMES:**Along the lines of those, 

### 23 sometimes if you don't control pests in your control plot, you 

### 24 may not have very good flowering and you could have poor control 

### 25 mortality. Poor growth of your colony and the control 

### 26 essentially is what your concerned about right, besides the

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1 possible interactive effects that you're adding a pest control 

### 2 measure. 

### 3 The other thing I wanted to add to Dr. 

### 4 Pettis's list. I hope I didn't miss it. It was the whole study 

### 5 had to be perhaps repeated in different locations and different 

### 6 places. Because if you do it one time in a same crop, I mean, 

### 7 it would be better if it could be repeated in multiple years or 

### 8 multiple sites and locations. The more times you could repeat 

### 9 it the better. You didn't really have that, right? Or did I 

### 10 miss it? 

### 11 **DR. JEFF PETTIS:**That's covered later on. In 

### 12 trying to say how robust the test would be, it's better to do it 

### 13 in several geographic areas. We haven't gotten to that. 

### 14 **DR. ROSALIND JAMES:**I apologize. You're 

### 15 right. That doesn't really address this question. 

### 16 **DR. DANIEL SCHLENK:**Okay. Yes, Dr. Pettis? 

### 17 Mr. Pistorius? 

### 18 **MR. JENS PISTORIUS:**Just a last comment on 

### 19 this. I think the uncertainties can be, to a large extent, 

### 20 avoided. Of course, it should be avoided that if you have a 

### 21 flowering crop where you do your tests that you apply no other 

### 22 insecticides. Also with fungicides because then you might run 

### 23 if that is not the purpose of the study into testing synergisms, 

### 24 but I think that actually should not be a problem. 

### 25 **DR. DANIEL SCHLENK:**Yes, Dr. Pettis? 

### 26 **DR. JEFF PETTIS:**Just again, back to both

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1 those comments. I think the need to have a crop in the control 

### 2 that's viable, that's producing and flowers and stuff is 

### 3 important, but you have to balance that. Maybe the best is to 

### 4 not use a target crop and use something like Phacelia that 

### 5 wouldn't need those treatments. But again, the best is to 

### 6 actually apply the formulated material to the crop of interest, 

### 7 but you have to take measures to make sure the control plots are 

### 8 not confounding but in good health. 

### 9 **DR. THOMAS POTTER:**Exactly, and I think 

### 10 those are all things that come into experimental design and I 

### 11 think are intended to, I guess, provide support for that process 

### 12 that was described in the White Paper. This is Tom Potter here. 

### 13 **DR. DANIEL SCHLENK:**That was Tom Potter. Any 

### 14 other strengths or limitations out there? Okay. Let's go ahead 

### 15 and move onto the next section, question 12. 

### 16 **MR. JOSEPH DECANT:**Joe DeCant, question 12, 

### 17 subpart (b) - please comment on the proposed modifications to 

### 18 the field study design elements presented in section 4.3.2 of 

### 19 the White Paper. 

### 20 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 21 **DR. JEFF PETTIS:**There will be some 

### 22 repetition throughout these things, but - so testing the 

### 23 formulation at the highest label weight is valid. I think 

### 24 that's a good idea. We agreed that plot location is a critical 

### 25 step in competing vegetation is an important consideration to 

### 26 ensure exposure to the target crop. The exposure achieved can

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1 accurately simulate the actual use of the pesticide, I think 

### 2 that's already been stated. 

### 3 The proposed studies are meant to stimulate 

### 4 worst case scenarios. This is true to the extent that the 

### 5 pesticide is applied when the highest label rate on the blooming 

### 6 crop. However, limited plot size may result in a less than 

### 7 worst case scenario in that these in fact are not going to be 

### 8 foraging solely on the target plot. 

### 9 Until the pesticide is in actual use and used 

### 10 on large acreage is you may not see the full extent of the 

### 11 exposure. So, that's kind of a limitation. 

### 12 I've already pointed out that plot size may 

### 13 vary depending on the attractiveness of the crop and that less 

### 14 attractive crops may necessitate having large field sizes. This 

### 15 is between controlled and treated plots. The exact distance 

### 16 should may be a minimum of three to six kilometers, but that 

### 17 distance has to be balanced to get some need to keep those crops 

### 18 in the same geographic and microclimatic conditions. So, there 

### 19 is a balance there between the need to keeping them in the same 

### 20 area and yet keep the distance such that you are not overlapping 

### 21 the foraging. 

### 22 We agree that balancing or equalizing of bee 

### 23 colonies is an important step to be taken in advance of placing 

### 24 the hives in the study site. Otherwise, colony variation 

### 25 between colonies is too great to see differences. So, colony 

### 26 equalization prior to placing the colonies into things is a very

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1 good step. 

### 2 We agreed that power analysis is a useful tool 

### 3 to determine the number of colonies needed to produce a 

### 4 statistically significant result. However, in practice often, 

### 5 plot size and replication are not such that we can use the power 

### 6 analysis and those studies aren't often done with high 

### 7 statistical power. We think the use of a power analysis to at 

### 8 least guide those studies should be used. 

### 9 The potential endpoints listed are useful to 

### 10 look at potential adverse effects. I have a couple of specifics 

### 11 that you list the returning of forager bees. I think you should 

### 12 be listing something more like the returning of bees over some 

### 13 time period. 

### 14 I would add that maybe we should add a 

### 15 consideration of the brood pattern, so you can analyze the 

### 16 consistency of the brood pattern by measuring 100 cell areas, 

### 17 and that's not listed on your list. So, there's a way to 

### 18 measure the consistency or solidness of the brood pattern. We 

### 19 have shown in other studies that that can be an accurate 

### 20 predictor of overall colony health. 

### 21 We concur that the duration of the field trial 

### 22 is going to be variable, and we dictate it by the endpoints of 

### 23 interest and also by the crop the products are sprayed on. 

### 24 We also concur that opening the hives can 

### 25 result in increased disturbance from possible introduction of 

### 26 variation within the results, increased queen loss, things like

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1 that increase spikes in adult bee mortality, but the hive 

### 2 manipulation shouldn't be minimized, but undue concerns about 

### 3 introducing these artificial effects. So, as long as the 

### 4 colonies in both plots are treated the same, I think you could 

### 5 minimize that. I don't think the disturbance factor is so 

### 6 great. So, hire a good beekeeper basically. 

### 7 And the last comment - most of these field 

### 8 studies are designed with use of honey bee colonies. So, you 

### 9 design a large field study and you put honey bee colonies in 

### 10 there. Certainly, you already have the field design there and 

### 11 the big problems with these are size of the plot and separation 

### 12 between control plots and treated plots. The use of things like 

### 13 Bombus or Osmia added to those plots would not incur a great 

### 14 deal of cost. Both of those genera fly shorter distances, so 

### 15 they would be more likely to stay within the field. 

### 16 So most of these Tier III studies are designed 

### 17 with honey bees in mind with the addition of Bombus or Osmia to 

### 18 those field studies could be telling in that both they fly 

### 19 shorter distances, they stay within the treated field and 

### 20 secondly, they have different life histories. So, you might see 

### 21 different effects by using two additional species. And that's 

### 22 all I have. 

### 23 **DR. DANIEL SCHLENK:**Thank you. Dr. Ostiguy? 

### 24 25 **DR. NANCY OSTIGUY:**I concur. 

### 26 **DR. DANIEL SCHLENK:**Dr. Potter?

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1 **DR. THOMAS POTTER:**I concur with everything 

### 2 that was said. I just wanted to add that I think there is some 

### 3 merit in value to possibly incorporating these studies with 

### 4 terrestrial field dissipation studies or plant residue decay 

### 5 studies. The more bang you can get for the buck obviously would 

### 6 be great. So, certainly there is potential for that type of 

### 7 integration here and certainly something to be contemplated and 

### 8 encouraged. 

### 9 **DR. DANIEL SCHLENK:**Dr. Tarpy? 

### 10 **DR. DAVID TARPY:**This is David Tarpy. I 

### 11 agree with Jeff's excellent real assessment there and really 

### 12 have nothing else to add except underscore the utility of the 

### 13 power analysis and that that's a very useful tool. 

### 14 **DR. DANIEL SCHLENK:**Other Panel members? Dr. 

### 15 James? 

### 16 **DR. ROSALIND JAMES:**I have a few things to 

### 17 add to the list. In terms of the honey bees, in terms of all 

### 18 bees, bees are like the rest of us and they are somewhat lazy 

### 19 and they will only fly as far as they have to generally. So, if 

### 20 you can keep the - if there is plenty of bee forage available, 

### 21 they are less likely to fly long distances and leave the 

### 22 treatment area. So, placing the bees in the middle of the field 

### 23 and making sure that there's lots of bloom and the design of the 

### 24 experiment will help. Like a lettuce field, you know, is not 

### 25 going to have a lot bloom in it. 

### 26 Then, going back to table 1, which I was in

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1 charge of table 1, so I'm going to go back to it. One of the 

### 2 things that we talked about was that the brood nest size would 

### 3 be an important measure. So for your list of measures, I would 

### 4 add brood nest size. That's something relatively easy to 

### 5 measure. 

### 6 Then, Dr. Pettis brings up a good point about 

### 7 other bees, and this is a good place now where we can move away 

### 8 from the honey bee being entirely a surrogate. Since you're 

### 9 doing a field trial, there are two things to consider. One, you 

### 10 could have a sample - there's a bee in front of me right now - 

### 11 you could just go out and do net sampling or pan traps, do some 

### 12 sort of sampling for what bees are already in the field and 

### 13 compare between control and treated or before and after. 

### 14 There, you can get your bee diversity measure 

### 15 in there and hit your third production goal, but also as risk 

### 16 managers, what is the crop being registered for. If it's being 

### 17 - the pesticide excuse me - if the pesticide is being registered 

### 18 for a crop in which something other than honey bees as a primary 

### 19 pollinator, then maybe the field trial ought to be conducted 

### 20 with that pollinator instead of honey bees. So, if it's being 

### 21 registered for alfalfa seed, you ought to be looking at alfalfa 

### 22 leaf cutting bee. If it's being registered for a tree crop, you 

### 23 could include something like one of the Osmia bees. So, those 

### 24 are my additions, thank you. 

### 25 **DR. DANIEL SCHLENK:**Any other input? Yes, 

### 26 Dr. Hunt?

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1 **DR. GREG HUNT:**I don't know if this was 

### 2 mentioned, but it's also important to do scouting to see what 

### 3 the bees are foraging on, to see if they're on nontarget crops 

### 4 and look at the pollen they're bringing in. 

### 5 **DR. DANIEL SCHLENK:**Okay. Anyone else? 

### 6 Okay. Apologies - I forgot to ask you guys on the last 

### 7 subquestion if you had any comments. If you have any comments 

### 8 on the last one and this one, any questions of clarification? 

### 9 Yes, Mr. DeCant? 

### 10 **MR. JOSEPH DECANT:**Joe DeCant, EFED. Maybe a 

### 11 clarifying question about the plot sizes - Dr. Pettis, you 

### 12 mentioned about the different crops given different 

### 13 attractiveness may require different size of the plots to be 

### 14 adequate in terms of the field study. Does the panel have any 

### 15 recommendations or thoughts on minimum plot sizes to be able to 

### 16 be able bound what we are looking at in terms of the field study 

### 17 and plot sizes, or a way to calculate how we're going to come up 

### 18 with the plot sizes or the field study related to the crop? 

### 19 **DR. JEFF PETTIS:**Jeff Pettis - on plot size, 

### 20 if you calculate the foraging range of an average colon, even 

### 21 conservative, and you even to a 1-hectare plot or a 5-hectare 

### 22 plot, the proportion of the total forage area that they could 

### 23 visit is really small. So, it's terrible to say, but you know, 

### 24 larger plots are better and more attractive and reducing 

### 25 competing vegetation. I would - this is maybe personal and 

### 26 maybe you can speak to this a bit more. Somewhere, 1 hectare is

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1 small, 5 hectares is starting to get, for me, is seemed like you 

### 2 would be getting in the realm of a reasonable plot size. What, 

### 3 are you thinking larger? I've seen some of these field studies 

### 4 where you can get replication. You don't end up having large 

### 5 fields, I don't know. 

### 6 **DR. DANIEL SCHLENK:**Okay. Who would like to 

### 7 go first? Dr. James? 

### 8 **DR. ROSALIND JAMES:**I was going to suggest 40 

### 9 acres for your plot size. I don't think that is ridiculous. 

### 10 And for seed - so the rules for certified seed and not getting 

### 11 cross pollination to keep your seed certified, I think, is 1 

### 12 mile. So, you know, that would be 1 mile between plots. 

### 13 **DR. DANIEL SCHLENK:**Mr. Pistorius? 

### 14 **MR. JENS PISTORIUS:**Well, I think this is one 

### 15 point where the EPPO needs some modification. Larger field 

### 16 sizes should be used and described in the old EPPO version, but 

### 17 I'm quite sure in the next version, there will be an update on 

### 18 this. 

### 19 I think we have to consider the different 

### 20 crops. For instance, if we compare maze and winter oilseed rape 

### 21 - for maze, I would personally go for larger field sizes then 

### 22 would be necessary for winter oilseed rape. It depends on the 

### 23 landscape where you do such studies. 

### 24 My personal experience, in former times when I 

### 25 was young, I conducted such studies a lot. We have a region 

### 26 where I come from and where I work where you could actually do

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1 peer trials, which I think give you the information that you 

### 2 need with winter oilseed rape of size of one hectare even. 

### 3 I don't know if that is appropriate and I'm 

### 4 not quite sure the agricultural structure is the same, but we 

### 5 had a discussion on this too, and it basically depends on the 

### 6 interpretation of effects and demonstration of exposure. I 

### 7 think, for instance, if you look at the number of foragers. 

### 8 Nowadays, we will say okay, maybe this one hectare field size 

### 9 used to be small for winter oilseed rape in former times. 

### 10 Nowadays, we say okay, yeah we want to go a little bit larger 

### 11 because you know in spring, it might be hard to track all the 

### 12 bees even if you really ensure that in a two or three kilometer 

### 13 circle, you have no relevant major flowering crops around. 

### 14 It may be different, for instance, if you do 

### 15 trials with Phacelia because the foraging density is a 

### 16 completely different one. You get, for instance in our 

### 17 condition, 5-OH foragers per square meter in winter oilseed rape 

### 18 and you get about 25 or 30 for Phacelia. So, I think in terms 

### 19 of these numbers, you may have a certain difference. I think 

### 20 with one or two hectares, you can start a very bee-attractive 

### 21 crop and ensuring that there is no cross foraging possible and 

### 22 landscape which does not provide alternative forage basically. 

### 23 You will not be able to invite completely, but no major nectar 

### 24 flow and no major pollen flow. Then you could start to get very 

### 25 good results with this. 

### 26 When you interpret it as a risk assessor,

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1 basically the effects that you get with care, that is a crucial 

### 2 point because we talked about if you have a controlled level of 

### 3 mortality of whatever 50. If you get 50 more dead bees, so 100 

### 4 dead bees after the application, let's assume that it's a 

### 5 pyrethroid. If you then careful as a risk assessor and say 

### 6 okay, I've got a concern. Then, it doesn't matter. You know, 

### 7 if you have five hectares or 50 hectares, the amount of bees 

### 8 that you will find from your colony is also limited. Because if 

### 9 you have the exposure maximized and if you basically achieve 

### 10 that the very large portion of your foragers visit this field 

### 11 and brings back this contamination - I know this is a crucial 

### 12 point - then you ensure that those have been well exposed. 

### 13 The number of foragers is limited to some 

### 14 extent, and you will not get completely different results if you 

### 15 have good exposure in smaller fields compared to when you have 

### 16 fields with 50 hectare. It has been the work of Helen Thompson, 

### 17 the colleague from the U.K. where they have actually addressed 

### 18 these concerns in studies with the pyrethroid in the U.K. where 

### 19 they had a field study of I think, one-half or two hectares of 

### 20 winter oilseed rape. Then they compared it to a landscape 

### 21 application with 200 hectares. The outcome was not different 

### 22 from the individual colonies. So, I think, you know, we all 

### 23 want that the bees are well exposed but maybe depending on the 

### 24 circumstances, but I think one or two hectares for flowering 

### 25 crops you're well off. Maybe for pollen and maze, you should 

### 26 need a little bit larger one.

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1 **DR. DANIEL SCHLENK:**Okay. First Dr. Pettis 

### 2 and Dr. Ostiguy. 

### 3 **DR. JEFF PETTIS:**Just to follow up, I would 

### 4 agree that one hectare maybe on the low end, but for crops such 

### 5 as Phacelia where you have high visitation rates and in areas 

### 6 where you don't have competition, they could be adequate. I 

### 7 would feel more comfortable with five hectare plots for general 

### 8 crops. 

### 9 You have to in all cases, trap incoming pollen 

### 10 and analyze that pollen to guarantee that their visiting the 

### 11 crop of interest, there is not too much competition, or if there 

### 12 is competition, it's equal between the control and the treated 

### 13 plot. So, there are things you can do to guarantee that you're 

### 14 getting some visitation or reasonable visitation to the plot 

### 15 size. It's just in practicality that large plots would be 

### 16 great, but they're just not usually feasible in most settings. 

### 17 So, five hectares may be - and then maybe down even as low as 

### 18 one hectare on very attractive crops. 

### 19 **DR. DANIEL SCHLENK:**Okay. Dr. Ostiguy? 

### 20 **DR. NANCY OSTIGUY:**This is actually where 

### 21 Tier III studies are of concern to me because I think that you 

### 22 get a fair amount of type II error. It helps knowing that the 

### 23 bees have actually foraged on what you want them to forage on, 

### 24 but our sizes of plots are too small. I think part of the 

### 25 difference between what Dr. James said and Dr. Pettis has to do 

### 26 with differences between the eastern U.S. and the western U.S.

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1 Your plot sizes in the west are going to be much larger 

### 2 generally. 

### 3 I was originally a westerner. That's sort of 

### 4 my vision also is a bigger area. I recognize though that here 

### 5 in the east, the size of each plot is smaller, but it does 

### 6 present a concern to me about type II errors. 

### 7 **DR. DANIEL SCHLENK:**Dr. James. 

### 8 **DR. ROSALIND JAMES:**This is Rosalind James. 

### 9 This occurred to me too the difference between I think even the 

### 10 midwest probably, although I have less experience with the 

### 11 midwest and the west. We are talking about, you know, 

### 12 three-quarters of the United States. If we throw the midwest 

### 13 and the west in there together, and especially something like 

### 14 California agriculture. California is wall-to-wall agriculture, 

### 15 and there is not a lot of margin places for the bees to hide out 

### 16 in. 

### 17 So if you have pesticide use, it's going to be 

### 18 solid and you're talking about cornfields in the midwest, it's 

### 19 going to be solid. There's not going to be other places for the 

### 20 bees to gain forage from. Typical irrigation unit in the west 

### 21 is 40 acres. If you're doing center pivot irrigation, I think 

### 22 it's 120 acres. So that's one farmer's field that is going to 

### 23 be very large. Now, so that's one reason why I say 40 acres, 

### 24 which is what about 12 hectares maybe. It's going to more 

### 25 because it's three to one. So about 12 hectares would be what 

### 26 we're talking about.

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1 But you could put more than one colony of bees 

### 2 in the middle of that. You could put four or so there or more. 

### 3 So, you could put more than one colony in the middle of that and 

### 4 it's somewhat pseudo-replication. So, your experimental design 

### 5 is going to be affected by that. 

### 6 **DR. DANIEL SCHLENK:**Dr. Hunt? 

### 7 **DR. GREG HUNT:**This is just a side note. 

### 8 Regardless of plot size, it may be a good standard practice or a 

### 9 practice to use sometime to just collect the incoming foragers 

### 10 and analyze them for pesticides to at least document that they 

### 11 were exposed. I don't know how good an idea it will give you of 

### 12 the total level of exposure, but some qualitative documentation. 

### 13 **DR. DANIEL SCHLENK:**Yeah, Dr. Pettis? 

### 14 **DR. JEFF PETTIS:**I think it's a good idea, 

### 15 Greg. I guess in the studies I've seen that trapping incoming 

### 16 pollen gives you a really nice measure of the level of exposure, 

### 17 you know, because you can trace it to the specific crop that 

### 18 you're growing. I was going to ask you, Dr. James, a question 

### 19 about - because let's say you can only have one plot of each. I 

### 20 mean, ideally, you have three or four plots of each type. There 

### 21 I think I would argue for small plot size. Take your 40 acres 

### 22 and divide it four ways into four 10 acre plots and get true 

### 23 plot replication because otherwise, we end up with just putting 

### 24 10 colonies on this one plot and using the colony as the source 

### 25 of replication. Ideally, you'd like to use the plot as the 

### 26 source of replication, but that's not always possible. So, I

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1 think that's some of the trade offs. 

### 2 **DR. DANIEL SCHLENK:**Okay. Yeah, Mr. 

### 3 Pistorius? 

### 4 **MR. JENS PISTORIUS:**I agree, but I think that 

### 5 actually comes in a later question. It would be desirable to 

### 6 have those in different environments, then you can actually 

### 7 cover part E of the question a little bit and do the 

### 8 representativeness. We do that even in our very tiny Germany. 

### 9 We ask for northern, eastern and southern or something like that 

### 10 when we test different environments. 

### 11 **DR. DANIEL SCHLENK:**Okay let's - you still 

### 12 have another comment? Yeah? Okay. Dr. James? 

### 13 **DR. ROSALIND JAMES:**Watching the clock, he 

### 14 wants to go home today. But maybe in the west we just have more 

### 15 land, and a 40 acre plot doesn't seem outrageous to me. I'm not 

### 16 back down. I think one hectare is way too small. Five - I 

### 17 don't know, your bees fly a long way. 

### 18 **DR. DANIEL SCHLENK:**So, let me make a 

### 19 recommendation instead of bantering back and forth. What's in 

### 20 the written documentation, you can agree to disagree and say 

### 21 that the positives for the small term plots and then you can 

### 22 document your justifications for the larger term plots. Yep, 

### 23 Dr. James? 

### 24 **DR. ROSALIND JAMES:**With my suggestion that 

### 25 you could sample for bee diversity within the plot, if your plot 

### 26 is too small, I don't think you can - sampling for bee diversity

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1 is not going to work as well because you're going to have the 

### 2 potential for influx from outside bees into your plot also. 

### 3 Unless you have immediate acute kill, you may not be able to 

### 4 tell that there's been an effect on bee diversity. So, if the 

### 5 plot's too small, that measure may not be effective for you. 

### 6 **DR. DANIEL SCHLENK:**Okay, which one? Dr. 

### 7 Pettis? 

### 8 **DR. JEFF PETTIS:**Okay. Back to the -- not 

### 9 dwelling on the larger plot size, but I think the point brought 

### 10 up by Dr. Pistorius was that if we have a chance of replication 

### 11 at the plot level, it's probably better across geographic 

### 12 regions. Then, you gather more information than doing it within 

### 13 one region. So, testing it in Kansas and Utah and Georgia would 

### 14 be preferable I think. 

### 15 **DR. ROSALIND JAMES:**The alfalfa seed isn't 

### 16 grown over all those areas. 

### 17 **DR. DANIEL SCHLENK:**Yes, Mr. Pistorius? 

### 18 **MR. JENS PISTORIUS:**Question to Dr. James. 

### 19 Did you say sampled bee diversity? Because I think that is a 

### 20 very complicated measure, because the bee diversity on such a 

### 21 crop would not be depending on this treatment basically, but how 

### 22 the population of different bee species was established before. 

### 23 If you cannot standardize this, then you cannot make basically 

### 24 an assumption later on how your treatment affected this bee 

### 25 diversity. 

### 26 **DR. DANIEL SCHLENK:**So, the question, I

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1 believe, for clarification was size, right? Is that basically 

### 2 what the clarification was? I mean, do you want more 

### 3 information with regard to other endpoints as well? 

### 4 **MR. JOSEPH DECANT:**This is Joe DeCant, EFED. 

### 5 The main question was about plot size, so I think the panel 

### 6 touched also on the number of hives per plot. So, we're talking 

### 7 about, you know, this brings in loading rate as well in terms of 

### 8 the number of hives per plot. I think that, you know, usually 

### 9 we're talking about three to four full-size hives at each plot. 

### 10 So, we're talking about hives of about 50 to 60,000 bees. 

### 11 Taking that into consideration in terms of the loading rates of 

### 12 the Panel, and the Panel's recommendations about plot size, if 

### 13 you also have a recommended loading rate, relating plot size to 

### 14 the number of hives per plot as well, that would be helpful. 

### 15 **DR. DANIEL SCHLENK:**Okay. That's not 

### 16 addressed further on in any of the questions? Okay. You might 

### 17 want to make a statement if you can at the present time so we 

### 18 can get it on the record. 

### 19 **DR. JEFF PETTIS:**Well, certainly with a small 

### 20 plot size, you could, by putting large colonies on the small 

### 21 plots, you could overwhelm the plot and there wouldn't be - but 

### 22 you still may get some representation, foraging rate and each 

### 23 colony would get some level of exposure. There is probably a 

### 24 formula out there that would, depending on the attractiveness of 

### 25 the crop, you could predict exactly how many colonies. It was 

### 26 kind of like a pollination, like how many colony spray do you

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1 need for almonds, almond crop, things like that. 

### 2 So, there probably are formulas out there. 

### 3 They're not at the top of my head right now. So, we can address 

### 4 that a bit in balancing the plot size versus the number of 

### 5 colonies. Even with and extremely small plot size, you're still 

### 6 - four colonies per plot would be a minimum because three is the 

### 7 minimum for any kind of replication. 

### 8 With honey bee colonies, one is going to die 

### 9 on you automatically. So, you have to start with four. I 

### 10 prefer to go up from that. But you will recognize that there 

### 11 could be an overloading effect and try to address that. 

### 12 **DR. DANIEL SCHLENK:**Okay. Yeah, Dr. Klaine? 

### 13 14 **DR. STEPHEN KLAINE:**If you put multiple hives 

### 15 on a single plot, isn't that pseudo-replication? 

### 16 **DR. JEFF PETTIS:**It is. 

### 17 **DR. ROSALIND JAMES:**This is Rosalind James. 

### 18 Or you could consider like a subsample. I mean, you want to put 

### 19 multiple hives on there because you might lose one for various 

### 20 reasons. 

### 21 **DR. STEPHEN KLAINE:**Then your replicate is 

### 22 your plot, not your hive. 

### 23 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 24 **DR. JEFF PETTIS:**But there is some discussion 

### 25 about that, because really you want to expose the bees and the 

### 26 unit that you're measuring is the colony. You're measuring a

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1 colony level response. So, in some sense, the colony is the 

### 2 replicate. But in reality, we would like to have even that 

### 3 colony replication exposed in multiple plots. That's a better 

### 4 design. 

### 5 **DR. DANIEL SCHLENK:**So, again, just letter E 

### 6 I think we talk about study design elements. So, if we can - 

### 7 yeah. Okay. 

### 8 **DR. ROSALIND JAMES:**I want to answer the 

### 9 question about how many bees per acre because I think I could 

### 10 give you a rough guideline of kind of a magic number, 3000 bees 

### 11 per acre. 

### 12 **DR. DANIEL SCHLENK:**That would be Dr. James. 

### 13 **DR. ROSALIND JAMES:**This is based on what we 

### 14 do with alfalfa leaf cutting bees, which are slightly smaller 

### 15 than honey bees and generally the growers put out really about 

### 16 5000 bees per acre. I'm sorry, 30,000 bees per acre. I will 

### 17 correct that. From, I think, 30,000 bees per acre is probably a 

### 18 good rule of thumb. That would be one colony per acre. 

### 19 **DR. DANIEL SCHLENK:**Just to remind the Panel, 

### 20 we're not here to try to get out quickly, we're just trying to 

### 21 focus the discussion to the questions that are being asked. So, 

### 22 that's what my job is, is to try to focus us. It's not to get 

### 23 us out early and get us home early, just to let you guys know 

### 24 that. We're just trying to focus the discussion to the question 

### 25 that's being asked. So, again, Mr. Pistorius you have another 

### 26 comment?

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1 **MR. JENS PISTORIUS:**Just a question for 

### 2 clarification basically, how much is 3000 bees per acre? 

### 3 **DR. ROSALIND JAMES:**30,000 per acre, so that 

### 4 would be 2.5 per acre. 

### 5 **MR. JENS PISTORIUS:**There are 2.5 acres in a 

### 6 hectare? Okay, thank you. Therefore there is also some 

### 7 recommendation on the minimal number of bees per square meter 

### 8 that should be there, and I think for Phacelia before you do 

### 9 your spray application, for Phacelia it's at least eight and I 

### 10 will have to look up the number. I think they said two to three 

### 11 for winter oilseed rape and other crops. 

### 12 **DR. DANIEL SCHLENK:**Okay. So, it sounds like 

### 13 several recommendations, which will be placed in the minutes. 

### 14 Dr. Pettis again, we are relying on you to put those in there 

### 15 and each panel member to provide their justifications for 

### 16 whatever numerical value they provide. It sounds like there is 

### 17 a little bit of disagreement. So, just make sure whatever 

### 18 number you come across, whether it's hectare size or bee number 

### 19 that there's a justification for the rest of the panel to weigh 

### 20 in on. Any other questions of clarification from the Agency for 

### 21 letter B? Okay. So, let's move onto letter C please. 

### 22 **MR. JOSEPH DECANT:**Joe DeCant, EFED, question 

### 23 12, subpart (c) - please comment on factors that should be 

### 24 considered in evaluating the biological significance of effects 

### 25 measured in full-field studies in relation to the proposed 

### 26 assessment endpoints and related protection goals.

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1 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 2 **DR. JEFF PETTIS:**Colony growth patterns is 

### 3 measured by brood production or adult bee population or robust 

### 4 measures of colony health and should be directly related to the 

### 5 protection goals or many of the protection goals. Similarly, 

### 6 things like colony mortality would be obviously biologically 

### 7 relevant, but you're unlikely to see these adverse direct 

### 8 effects. The effect on queen survival and queen replacement 

### 9 would be of biological significance as it has implications for 

### 10 long-term colony survival and may not be accounted for if test 

### 11 duration is only of a few weeks or months. 

### 12 Over winning success may be the real ultimate 

### 13 test for colony survival, but it's unlikely to be realized 

### 14 unless the exposure period is closer to the fall and you can 

### 15 control for factors outside the test period when colonies can 

### 16 suffer from parasitic mites and things like that. 

### 17 Adult bee mortality is a good biological 

### 18 indicator as are foraging rates in adult bee longevity. Each of 

### 19 these could be good indicators of adverse effects, but effects 

### 20 could be transitory in nature, and thus extrapolation to the 

### 21 colony level impacts is not necessarily direct. So, you may 

### 22 measure those effects, but how they affect total colony survival 

### 23 is not always one to one. 

### 24 Taken together any effects noted in the above 

### 25 endpoints could be used in the risk assessment decision making 

### 26 even if each endpoint alone was not significant, that is taking

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1 the total things measured and looking at those even if they 

### 2 weren't significant, the weight of impacts could be taken into 

### 3 consideration as well as Tier I and Tier II testing. 

### 4 Colonies are complex social organisms, and 

### 5 thus several endpoints measured may be more appropriate than 

### 6 relying on a single endpoint alone. 

### 7 Then finally, disease in pest levels and 

### 8 worker longevity, foraging rates and other endpoints that are 

### 9 measured are less robust, but they should be considered and they 

### 10 should be actually equalized as far as we can at the beginning 

### 11 of each study. 

### 12 Any adverse effects noted in things such as 

### 13 foraging rates or adult bee longevity during the study should be 

### 14 noted and taken into consideration with any evidence from Tier I 

### 15 or II testing. So, again, just what effects seen in Tier III 

### 16 testing should be looked at in context of anything gathered in 

### 17 Tier I or Tier II. Those were the comments that I had. 

### 18 **DR. DANIEL SCHLENK:**Okay. Dr. Ostiguy? 

### 19 **DR. NANCY OSTIGUY:**I concur. 

### 20 **DR. DANIEL SCHLENK:**Dr. Potter? 

### 21 **DR. THOMAS POTTER:**I concur. 

### 22 **DR. DANIEL SCHLENK:**Dr. Tarpy? 

### 23 **DR. DAVID TARPY:**I agree as well. 

### 24 **DR. DANIEL SCHLENK:**Other Panel members? 

### 25 Okay. Any questions of clarification? All right. We're going 

### 26 to move on to letter E then. Oh D, sorry.

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1 **MR. JOSEPH DECANT:**Joe DeCant, EFED, question 

### 2 12, subpart (d) - please comment on factors and methods that 

### 3 should be considered when extrapolating observed effects at the 

### 4 colony level in semi-field and field studies to those expected 

### 5 to occur in the environment, for example spatial and temporal 

### 6 scale of exposure, hive management practices, presence of 

### 7 multiple chemical and nonchemical stressors, et cetera. 

### 8 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 9 **DR. JEFF PETTIS:**In Tier II and Tier III 

### 10 studies, they cannot possible incorporate all seasonal and 

### 11 colony management practices that may occur, or changes in 

### 12 response to the colony due to seasonality changes in colony 

### 13 growth. But if testing is conducted in the normal growing 

### 14 season, a certain level of confidences can be had and the 

### 15 results generated. 

### 16 So, testing whatever the crop of interest is 

### 17 would normally be grown and then using colonies during that test 

### 18 period would be ideal. If you want to do a test and you do it 

### 19 outside, what would normally be expected as far as seasonality, 

### 20 then you could have less confidence in the results. 

### 21 As has been stated earlier, using colonies in 

### 22 either field or semi-field, they should be uniform in strength 

### 23 and as disease free as possible at the start of the study to 

### 24 avoid confounding effects. This is just a personal note - I 

### 25 would tend to skew towards smaller colonies, although there is 

### 26 some discussion in the White Paper about larger colonies being

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1 preferred. I think well-balanced, small colonies would give you 

### 2 reliable results and are meaningful biologically. 

### 3 The seasonal nature of colony growth and the 

### 4 importance of colony preparation for winter, extra stores and 

### 5 honey and things that gathered, should be taken into 

### 6 consideration if we're going to try to do a study where we're 

### 7 going to look at overwintering. So, the exposure period, if 

### 8 you're going to do an overwintering study, exposure period 

### 9 should be in the summer or early fall in advance of that to 

### 10 minimize the amount of time that those colonies are followed 

### 11 where we have less control of the variables. 

### 12 In Tier III studies, the longer the study 

### 13 continues or the more variables that will come into play, even 

### 14 if you've done a good job of equalizing the colonies at the 

### 15 beginning of the study relative to pest and disease, you're 

### 16 going to have other outside influences, such as other crops that 

### 17 they may visit in the interim, additional pesticide exposure, 

### 18 things that can influence the results of trying to follow these 

### 19 Tier III studies for longer periods of time. 

### 20 One way to approach this is to take colonies 

### 21 from the treated plots and the control plots and move them into 

### 22 a common area that is away from urban or agricultural settings 

### 23 and try to put them in an as clean and pristine environment as 

### 24 possible to avoid some of these confounding effectives. 

### 25 Lastly, there could be some consideration of 

### 26 whether a crop to be tested is a minor or major crop. If it is

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1 a major crop, then even the test scenarios under Tier III may 

### 2 not adequately reflect the ultimate use of the product in such 

### 3 that these may be exposed more widely. I'm thinking of crops 

### 4 like soy beans. So, if you test small plot size and you draw a 

### 5 conclusion and then the products in use are widespread, the 

### 6 number of acres in things like soy beans may make in reality, 

### 7 the exposure levels that bees receive in the real world even 

### 8 greater, and that would be during the test period. So, those 

### 9 are some of the ideas that we have. 

### 10 **DR. DANIEL SCHLENK:**Dr. Ostiguy? 

### 11 **DR. NANCY OSTIGUY:**I concur, but I'd also 

### 12 like to add that one of the issues that I have with Tier III 

### 13 studies is that our goal is to try to approach what are the 

### 14 ecological conditions that will occur during use of the 

### 15 material, yet we are at the same time trying to eliminate 

### 16 confounders. The confounders are part of the reality of the 

### 17 ecological conditions for how we use honey bees. So, our type 

### 18 II errors, we're in many ways by our design, increasing the 

### 19 chance that we're going to find the type II error. 

### 20 On the other hand, we have to control for the 

### 21 confounders, otherwise you don't have repeatability in your 

### 22 studies. So, this probably goes in at an evaluation level above 

### 23 the type II study itself, but there is a very severe limitation 

### 24 to how good the date actually are for the ecological conditions 

### 25 we see. 

### 26 **DR. DANIEL SCHLENK:**Thanks. Dr. Potter?

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1 **DR. THOMAS POTTER:**I concur with what Jeff 

### 2 and Nancy had stated. I have one thing to add. It comes in the 

### 3 form of a question, and also from long years of experience doing 

### 4 other types of studies, field studies, and that is the question 

### 5 of multiple year or studies that have to be conducted over 

### 6 multiple years to derive some sense of how climatic variables 

### 7 and other may strongly influence results. So, I do think that's 

### 8 been addressed here very effectively. Probably an appropriate 

### 9 topic for discussion. 

### 10 **DR. DANIEL SCHLENK:**Okay. Dr. Tarpy? 

### 11 **DR. DAVID TARPY:**My comments have all ready 

### 12 been made by the others. 

### 13 **DR. DANIEL SCHLENK:**Okay. Dr. Pettis? 

### 14 **DR. JEFF PETTIS:**To the question raised by 

### 15 Dr. Potter, I think it's listed here in the last set of 

### 16 recommendations. The idea of testing in several geographic 

### 17 locations may get at that a bit. It certainly doesn't account 

### 18 for all weather and possibilities, but if you do a test or a 

### 19 series of tests in several geographic locations, you have a bit 

### 20 more confidence that the results represented the real world. 

### 21 **DR. THOMAS POTTER:**I agree, but I don't with 

### 22 that provisionally. I don't think that simply looking at 

### 23 different geographic regions addresses the issue of local 

### 24 climate variability and the potential confounding effect on 

### 25 results. 

### 26 **DR. DANIEL SCHLENK:**That was Dr. Potter. Mr.

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1 Pistorius? 

### 2 **MR. JENS PISTORIUS:**Dr. Potter also mentioned 

### 3 the studies conducted over several years and the longer you 

### 4 conduct such a study, the more you get influence by the 

### 5 beekeeper and less by the pesticide. What you can do is for the 

### 6 environmental conditions, what is done in Europe for instance, 

### 7 if there were no concerns that stopped the registration that we 

### 8 ask for monitoring in the real world. So, you follow some bee 

### 9 colonies over a while. Nevertheless, the longer you do it with 

### 10 exactly the same colonies, treating every colony alike, the more 

### 11 get an influence of the beekeeping and of individual factors. 

### 12 Okay sorry. The longer you do it basically with the same 

### 13 colonies, the more you get an influence of other factors than 

### 14 pesticides I think. 

### 15 **DR. DANIEL SCHLENK:**Dr. Ostiguy? 

### 16 **DR. NANCY OSTIGUY:**I agree. This is one of 

### 17 the reasons that I would do a replicate study versus following 

### 18 the same colonies over years. Certainly, our experience with 

### 19 the large CAP study, keeping colonies beyond even a year can be 

### 20 a challenge. 

### 21 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 22 **DR. JEFF PETTIS:**I will just do a followup. 

### 23 I think your suggestion of using colonies that are 

### 24 representative of the real world is a good one. It kind of gets 

### 25 to Dr. Berenbaum's thing, that in reality, there are sick bees 

### 26 out there. They're not perfectly healthy. I guess I would

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1 argue for reasonably healthy colonies that are uniform in 

### 2 nature, as disease free as possible, but that's why I would 

### 3 argue for the small 10 to 20,000 bee colonies that you might be 

### 4 more likely to see an effect. And you also don't have this 

### 5 loading issue of putting 60,000 adult bee colonies in there. 

### 6 So, I would opt for smaller, well balanced colonies that are in 

### 7 reasonable health. 

### 8 **DR. DANIEL SCHLENK:**Any other comments? 

### 9 Okay. Let me go to the Agency. Any questions of clarification 

### 10 for letter D? Good. Okay. Let's move onto the last 

### 11 sub-question there, letter E. 

### 12 **MR. JOSEPH DECANT:**Joe DeCant, EFED, question 

### 13 12, subpart (e) - a number of study design elements are 

### 14 discussed in section 4.3.5 of the White Paper. However, even in 

### 15 the best designed studies, there can be confounding effects 

### 16 which can limit the utility of these studies in risk assessment. 

### 17 Please comment on factors that should be considered in 

### 18 determining the utility of field studies for pesticide risk 

### 19 assessment, including a discussion of the representativeness of 

### 20 a study for a National Level assessment, that is the pesticide 

### 21 may be used anywhere in the United States and its territories. 

### 22 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 23 **DR. JEFF PETTIS:**Again, kind of reiterating 

### 24 some of the things that were brought our earlier. If the 

### 25 pesticide is to be used on a widely planted crop common across 

### 26 the U.S., then testing should be representative and should be

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1 done in several environments across the U.S. for it to 

### 2 representative of a valid study. 

### 3 The ideal test would be using the formulated 

### 4 product on the crop of interest. In other words, definitely 

### 5 using it on the exact crop, not on a crop such as Phacelia, and 

### 6 with the highest level of attraction. So if you had multiple 

### 7 crops, maybe using it on oilseed rape, whatever the most 

### 8 attractive crop would be appropriate. 

### 9 If visitation to the crop is confirmed by 

### 10 analyzing pollen and you still see no adverse effects, then I 

### 11 think you could have some level of confidence that maybe you're 

### 12 not producing a type II error that you have good visitation, 

### 13 crop is sprayed at a high rate, then I think you can have some 

### 14 level of confidence that the study actually did give you a 

### 15 reasonable result. 

### 16 The design element, the plot size and plot 

### 17 separation, we've already gone over. I think they're critical 

### 18 in designing it. In an appropriate study, the identification of 

### 19 incoming pollen during the exposure period is probably the 

### 20 single best measure to give you some level of confidence that in 

### 21 fact, the exposure has occurred and additional residue analysis 

### 22 of that pollen can confirm that. 

### 23 I don't know how well that addresses all of 

### 24 the aspects in E, but maybe they can come out in the discussion. 

### 25 So, that's what I had. 

### 26 **DR. DANIEL SCHLENK:**Dr. Ostiguy?

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1 **DR. NANCY OSTIGUY:**I concur. 

### 2 **DR. DANIEL SCHLENK:**Dr. Potter? 

### 3 **DR. THOMAS POTTER:**I concur. What this 

### 4 sounds like is an opportunity to do some field work in Hawaii. 

### 5 So, that sounds good. I do want to add a couple of points here, 

### 6 and I think that again, it comes back to somewhat of what I said 

### 7 as a response in the prior question is that we need to very 

### 8 carefully look at climate and make sure that we're taking 

### 9 climate into account in terms of its impact on bee populations, 

### 10 and I'm talking about local climate, you know, in terms of equal 

### 11 regions within the country as well as variations in climate and 

### 12 certainly among other things that's year to year, and also of 

### 13 course issue surrounding climate change. 

### 14 So, you know, in designing these studies, you 

### 15 know I guess we need to put our thinking caps on and kind of 

### 16 work our way through some of these issues related to climate as 

### 17 well as geography, et cetera, but you know I generally agree 

### 18 with Jeff and unfortunately don't have the easy answer. 

### 19 **DR. DANIEL SCHLENK:**Dr. Tarpy? 

### 20 **DR. DAVID TARPY:**I agree with the comments 

### 21 that have been said all ready as well, and underscore the 

### 22 utility of ground truthing and ensuring that the bees are 

### 23 actually exposed and are foraging on the target crop for a Tier 

### 24 III study to come back with negative results that does not 

### 25 demonstrate that the bees were, in fact, foraging on that crop 

### 26 is not very useful, and that introduces that increased risk of

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1 type II errors. So, that's one way to really kind of 

### 2 standardize across these that the bees are actually doing what 

### 3 they're thought to be doing. 

### 4 **DR. DANIEL SCHLENK:**Other Panel members? Dr. 

### 5 James? 

### 6 **DR. ROSALIND JAMES:**I would just add, make 

### 7 sure you have irrigated and non-irrigated crops because humidity 

### 8 probably could play a big role and still would call for some 

### 9 measure of bee biodiversity in the field. 

### 10 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 11 **DR. MAY BERENBAUM:**This may be unanswerable, 

### 12 but hearing the requirements for plot sizes, you know, they're 

### 13 sufficiently large that are free of pesticide contamination that 

### 14 are in reasonably undisturbed areas that are replicated year to 

### 15 year to accommodate climatic variation. I live in central 

### 16 Illinois. I don't know that there are any places to put ideal 

### 17 experimental plot that isn't one way or another compromised. 

### 18 So, I don't know how to factor this in, but is this a 

### 19 limitation? I don't know. 

### 20 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 21 **DR. JEFF PETTIS:**You raise a good point, Dr. 

### 22 Berenbaum. I think in hearing the German situation, they have 

### 23 valleys in which there are heavily forested areas. They go in 

### 24 and plant specific plots up and down those different valleys. I 

### 25 think in large parts of the U.S. that's impossible and you have 

### 26 almost at any time the year, you have some competition. One

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1 thing I think you can do is look at early spring or summer where 

### 2 maybe there is less - there's other crops that are maybe there, 

### 3 but they're not attractive. Your planting them and hopefully if 

### 4 you're plots are not too far apart, you still got whatever the 

### 5 other crops of interest - or not crops of interest, the other 

### 6 competing crops are equal across that. Again, trapping incoming 

### 7 pollen is a way to measure the amount of visitation to your 

### 8 target crop, things like that. But there are huge obstacles in 

### 9 setting up decent plot size and then reducing competing 

### 10 vegetation. 

### 11 Just one other comment while I've got the mic 

### 12 - I think it's really important if we're looking at a product 

### 13 that is going to be used, I will say for a major use versus a 

### 14 minor crop. If we make an error on a major use and we're going 

### 15 to license something for use on a major crop, I think there is 

### 16 more responsibility there than if we make a mistake on a minor 

### 17 crop use, where the bees may or may not be attractive to the 

### 18 minor crop but it's not in widespread plantings. 

### 19 Major crops where things are being used, you 

### 20 know, in like you say the Midwest is a good example. Mistakes 

### 21 there where we don't adequately test and field test them, then 

### 22 those should be of concern. 

### 23 **DR. DANIEL SCHLENK:**Mr. Pistorius? 

### 24 **MR. JENS PISTORIUS:**I have to add I don't 

### 25 live anymore in the hilly areas. It is totally flat where I 

### 26 live now. Even there, some contract labs are situated and you

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1 can still do good tests also in flat landscapes, but we are in 

### 2 the same situation. It's just a matter of distance between 

### 3 fields. You may have to choose a little bit larger distances 

### 4 when it's hilly, but the main thing is the plot isolation. 

### 5 **DR. DANIEL SCHLENK:**Dr. James? 

### 6 **DR ROSALIND JAMES:**I think in the west this 

### 7 is less of a problem where the majority is agriculture. 

### 8 Two-thirds of American agricultural products come out of the 

### 9 state of California. And I have done large scale field 

### 10 experiments with farmers and you can get replicates in one year. 

### 11 Also, a lot of its desert, so you only have crop growing where 

### 12 there is irrigation and you don't have the much competing forage 

### 13 and forest and things like that, depending on where you do it. 

### 14 So, maybe a large number of these tests just need to be done in 

### 15 the west. 

### 16 **DR. DANIEL SCHLENK:**Any other comments? 

### 17 Okay. We go to the Agency. Mr. DeCant, you have any questions 

### 18 of clarification? 

### 19 **MR. JOSEPH DECANT:**Joe DeCant, EFED. I have 

### 20 one question of clarification. So, the Panel mentioned about 

### 21 the field studies and the design of the field studies and making 

### 22 sure that we have adequate plot sizes and looking at the 

### 23 chemicals under good agricultural practices, but at the same 

### 24 time, the Panel is recommending looking at foraging. So, part 

### 25 of this speaks to the study design and even if we have adequate 

### 26 study design, then we also need to ensure that the bees are

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1 foraging. So, my question to the panel is do you have any 

### 2 recommendations or thought on what would be considered an 

### 3 adequate level of foraging, either in the case of if we identify 

### 4 a difference between the control and the treatment in terms of 

### 5 where they're foraging or just an overall foraging and what we 

### 6 would consider as adequate. 

### 7 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 8 **DR. JEFF PETTIS:**I'll take a stab at that. 

### 9 Knowing that you can't control - I mean, you've got a wide array 

### 10 of options out there to forage on, just as a ballpark figure, if 

### 11 50 percent of the pollen was coming back from the target crop, I 

### 12 would think that would be good exposure. Anything above that is 

### 13 even better. And then, are you interested in foraging rate? 

### 14 Are you interested in something about foraging rate - I think 

### 15 just determining that they are foraging on the crop of interest 

### 16 and using the incoming pollen and the indentifying of that 

### 17 pollen such that you have some assurance that a reasonable 

### 18 amount of the exposure has been undertaken. For lack of a 

### 19 better - off the top of my head, 50 percent of the pollen, you 

### 20 know you're getting 50 percent of their diet from that target 

### 21 crop. With Dr. James's 48 acres, that would be easy to do, 

### 22 right? Acres, yeah. I don't think you want approach 100 

### 23 percent. You're just not going to get there, but anything 50 

### 24 percent or greater of the incoming pollen would indicate a 

### 25 reasonable amount of foraging on the target crop. 

### 26 **DR. DANIEL SCHLENK:**Let me just go through

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1 their main ones. Dr. Ostiguy, do you have a number, percentage? 

### 2 3 **DR. NANCY OSTIGUY:**No, I don't think there 

### 4 really - I don't know of any data that actually gives us that 

### 5 information. I'm just still back to my problem with type II 

### 6 errors. 

### 7 **DR. DANIEL SCHLENK:**Okay. Dr. Potter? 

### 8 **DR. THOMAS POTTER:**Getting back to what Dr. 

### 9 Berenbaum said about Illinois, certainly that's the case in some 

### 10 other parts of the country where you have the landscape 

### 11 literally heavily disturbed, but also covered with vast wasps of 

### 12 crops of interest. So, in the context of these studies, I'm 

### 13 trying to get my head around how you do pollen counts or pollen 

### 14 typing and are assured that that's indeed from the pesticide 

### 15 treated crop. So, I guess I'm asking a question here and maybe 

### 16 Dr. Pettis has some thoughts on that. 

### 17 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 18 **DR. JEFF PETTIS:**Obviously, you couldn't if 

### 19 you were treating canola. You couldn't have competing canola 

### 20 fields because they would be confounding. So, whatever your 

### 21 target crop that you're planting would need to be unique. So, 

### 22 you would have to be able to identify, and then you do analyze 

### 23 that 50 percent of the incoming pollen is from canola, and you 

### 24 analyze the pesticide level of that versus control to see if 

### 25 there's been crossing between the plots. It's fairly straight 

### 26 forward, and I agree that Midwest, it would be very difficult.

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1 I'm thinking desert, Southwest and these nice big circles. When 

### 2 you fly out west and you see these nice big circles, those are 

### 3 idea places of to do that. 

### 4 **DR. THOMAS POTTER:**Yeah, I brought that up 

### 5 because I don't - that is what I wanted to capture in the sense, 

### 6 in our notes here. This is Tom Potter. 

### 7 **DR. DANIEL SCHLENK:**Thanks. Dr. Tarpy? 

### 8 **DR. DAVID TARPY:**This is Dave Tarpy. You 

### 9 want a percentage from me or just to follow up on that? 

### 10 **DR. DANIEL SCHLENK:**Well, I think the EPA is 

### 11 wanting a measure of foraging success. 

### 12 **DR. DAVID TARPY:**So, to Dr. Pettis's point 

### 13 then on that, I think 50 percent is probably a good target. I 

### 14 mean in some cases, that might be high. I don't know if there 

### 15 is a magic number as far as percentage of returning pollen 

### 16 foragers with target crop pollen on their corbiculae, right? 

### 17 Because there are a lot of target crops where they are not 

### 18 pollen bearing at all and they are not going to be collecting 

### 19 any pollen, right? So, that may be difficult. 

### 20 What I was referring to as far as ground 

### 21 truthing was actually doing things like transect walks or what 

### 22 Dr. James was saying of the pan traps and actually collecting 

### 23 bees in those traps to verify that bees are actually in the 

### 24 field of interest. Actually, doing transects up and down the 

### 25 rows of that particular crop and visibly verifying that honey 

### 26 bees and other are foraging on those flowers. Therefore,

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1 putatively coming into contact with and being exposed with the 

### 2 chemical in question. 

### 3 There are many ways to get to that, and it's 

### 4 probably crop dependent as to the best measure of verifying that 

### 5 honey bees are actually in the target crop. 

### 6 **DR. DANIEL SCHLENK:**Okay. Other panel 

### 7 members? Dr. Hunt? 

### 8 **DR. GREG HUNT:**Yeah, these studies are so 

### 9 hard to set up. I don't know the answer to the question, other 

### 10 than to say that you ground truth, you measure the pollen, 

### 11 proportion, whatever it is and report it, and what was the other 

### 12 thing I was going to say - oh yeah, foraging rate, it depends on 

### 13 if what you're applying is expected to be like a foliar 

### 14 application and it's expected to be a burst of time point. Then 

### 15 you would want to take the foraging rate that day. 

### 16 **DR. DANIEL SCHLENK:**Okay. Any other 

### 17 comments? Oh sorry, Mr. Pistorius? 

### 18 **MR. JENS PISTORIUS:**Well, depending on the 

### 19 crop, it's very different. For instance, Phacelia, you can do 

### 20 very easily an optical differentiation because it's a purple 

### 21 pollen. Almost no other crop has a purple pollen. You can 

### 22 easily count by the numbers of pollen grain for that color. 

### 23 That is Phacelia pollen, if you have ensured that there are no 

### 24 other Phacelia crops around, which I assume no. It may be more 

### 25 difficult with, for instance, canola, because at that time of 

### 26 blooming probably the same year, you get a few other flowers or

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1 some other flowers in the surrounding, which also have a 

### 2 yellowish color. So, you have to do pollen analysis. 

### 3 Then what I still think is a very good measure 

### 4 is foraging, the number of bees foraging on that specific crop. 

### 5 For us for our conditions, we have by experience, we have 

### 6 minimum numbers. For instance, for winter oilseed rape, if we 

### 7 know that if it's below two or three foragers per square meter, 

### 8 it is not valid because also depends on the bee size. For 

### 9 Phacelia, you can say that normal is at least 12 to 25. I think 

### 10 those numbers may also - I mean we would have to look at other 

### 11 crops to find certain numbers. I mean for a tree, it is 

### 12 probably more difficult to estimate, but our possible estimates 

### 13 to ensure that the bees have actively been foraging on the crop. 

### 14 15 **DR. DANIEL SCHLENK:**Yes, Dr. Fefferman? 

### 16 **DR. NINA FEFFERMAN:**Sorry, I just have a 

### 17 question and listening to all this. Hopefully, this question is 

### 18 really stupid, but it gets back to something Dr. Berenbaum 

### 19 mentioned about choice. I'm wondering if we separated the 

### 20 exposure route from actual ingestion versus contact if we should 

### 21 worry at all about landing rates on crops that are then rejected 

### 22 as forage, if that's a contact exposure without the concomitant 

### 23 ingestion exposure. 

### 24 **DR. DANIEL SCHLENK:**Again, I think we're kind 

### 25 of trying to answer a design question here with this. Okay. 

### 26 Anybody want to chime in? Yeah, Dr. Hunt?

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1 **DR. GREG HUNT:**Well, if I understand the 

### 2 question, you're talking about bees visiting a flower and then 

### 3 rejecting it. Once they reject that flower, they've all ready 

### 4 landed on it. I think you'll get this from the scouting. You 

### 5 know, if they continue to reject, they will continue visiting 

### 6 flowers of the same kind. But pretty soon, they're going to 

### 7 look for something better. 

### 8 **DR. NINA FEFFERMAN:**This is Nina Fefferman 

### 9 again. This is basically my thought in response to Dr. Tarpy's 

### 10 idea of watching who's flying where and having that mismatch 

### 11 with what they're doing. So yeah, I agree over time if they're 

### 12 going back and communicating that it wasn't a good source and 

### 13 they have to go elsewhere, yeah they just won't keep visit the 

### 14 area. But what you're locally doing is walking a transect and 

### 15 just looking, let's say early in the morning for where scout 

### 16 foraging is going. That might be a real disconnect in what's 

### 17 happening. 

### 18 **DR. DANIEL SCHLENK:**Okay. Does that answer 

### 19 your question in regards to foraging? Any other questions of 

### 20 clarification? Okay. Let's go ahead and move on to 13. 

### 21 **DR. THOMAS STEEGER:**This is Tom Steeger, 

### 22 question 13A please comment on the use of data on individual 

### 23 bees to transition to higher tier studies given that the Tier I 

### 24 studies focus on survival as the primary measurement endpoint 

### 25 although additional endpoints may be forthcoming as test designs 

### 26 continue to develop.

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1 **DR. DANIEL SCHLENK:**Our lead discussant is 

### 2 Dr. Tarpy. 

### 3 **DR. DAVID TARPY:**This is Dave Tarpy, NC 

### 4 State. So, many of these points or at least for this 

### 5 sub-question, I think have been articulated all ready. For the 

### 6 sake of time and just to not have to recapitulate them all over 

### 7 again, we'll just refer to those previous questions. So, this 

### 8 is starting to funnel up a lot of our discussions from yesterday 

### 9 and today on this and starting to distill them. 

### 10 In general, much of those discussions, it 

### 11 seems pretty clear that there isn't this perfect proxy of Tier I 

### 12 and lower tiers that really capture a lot of the complexity of 

### 13 the upper tiers. This is especially true in social systems like 

### 14 in honey bees. So, additional care needs to be taken in trying 

### 15 to equivocate these different tiers. 

### 16 So, because of that, I think a lot of what 

### 17 we've been talking about in question 8, question 11 in 

### 18 particular about blurring the lines between the Tier I, II and 

### 19 III in using groups of individuals rather than individual bees, 

### 20 not solitary bees. But to incorporate bioassays that test the 

### 21 social unit will go a real long way to addressing some of this. 

### 22 Certainly not completely, but it really does help to capture the 

### 23 biological reality of looking at colony and group phenotype 

### 24 rather than just individual phenotype. 

### 25 With of course the exception of queens and 

### 26 drones, where looking at individual phenotypes on their

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1 reproductive potential is something that would be obviously 

### 2 worthwhile. We also agree as the discussants that mathematical 

### 3 modeling will be able to help hopefully tie a lot of these types 

### 4 of aspects together, particularly if the same phenotypes are 

### 5 measured at these different levels of reductionistic to larger 

### 6 scale types of experimental designs. So if the same phenotypes 

### 7 can be measured at those different increasing levels being able 

### 8 to kind of verify them across those different levels of 

### 9 biological organization would be very powerful means of 

### 10 verifying that and using mathematical modeling to have 

### 11 predictions from one level to another. 

### 12 We also, and Dr. Berenbaum can extrapolate on 

### 13 this if she wants, but that data on the individual pupae can be 

### 14 very informative at lower levels to higher levels. That doing 

### 15 Tier I bioassays on contact exposure on individual pupae, that 

### 16 is likely to have very strong penetrants to colony-level 

### 17 phenotype compared to some of the other more nebulous ones like 

### 18 we've been talking about of subtle behavioral differences, which 

### 19 again can have very important biological relevance, but much 

### 20 more difficult to capture and to translate from one level to 

### 21 another. 

### 22 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 23 **DR. MAY BERENBAUM:**What I was going to say 

### 24 was said, so I concur. 

### 25 **DR. DANIEL SCHLENK:**Great. Dr. Pettis? 

### 26 **DR. JEFF PETTIS:**I concur.

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1 **DR. DANIEL SCHLENK:**Other Panel members? 

### 2 Must be getting late in the day. Oh, sorry, Mr. Pistorius? 

### 3 **MR. JENS PISTORIUS:**Well, I just want to make 

### 4 a comment, for instance, on the IGRs. I may have made this 

### 5 comment all ready. I think for IGRs for instance, that the 

### 6 current Tier I studies as they are designed, also taking into 

### 7 account that there's a seven-day Aupinel method available soon, 

### 8 we hope. Unless the Aupinel method is soon also extended to the 

### 9 pupation and the hatching phase, that just looking at the 

### 10 individual mortality data of adult bees as measured by acute 

### 11 oral or contact toxicity, we would miss the effects of IGRs. 

### 12 In principle, I agree that this data on the 

### 13 individual bees, the transition to higher tier tests is a 

### 14 reasonable approach given some limitations, but I still think 

### 15 that there must be some flexibility for the risk assessor to 

### 16 identify, for instance, like the IGRs, the need for other 

### 17 information like the mode of action go to higher tier tests 

### 18 immediately or in addition. So, saying that for the risk 

### 19 assessor, there is a need that we don't have a calculation 

### 20 somewhere. This is limiting the option to go into higher tiers 

### 21 because currently there would be, or if you say okay, no trigger 

### 22 bridge by what we know. Some things could be missed. 

### 23 **DR. DANIEL SCHLENK:**So, it sounds like you 

### 24 would like at qualitative component to that sort of decision 

### 25 process there. Any other comments on letter A? Okay. Let's 

### 26 move forward to letter B.

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1 **DR. THOMAS STEEGER:**This is Tom Steeger, 

### 2 question 13B - please comment on the derivation of the Level of 

### 3 Concern, that is the LOC of 0.4 and the extent to which is 

### 4 serves as an appropriate screen to transition to higher tiers of 

### 5 testing and refinement. 

### 6 **DR. DANIEL SCHLENK:**Dr. Tarpy? 

### 7 **DR. DAVID TARPY:**So this is Dave Tarpy. The 

### 8 discussants on this particular sub-question would also like to 

### 9 hear from many of the toxicologists on the panel to provide 

### 10 their insights since none of us really are toxicologists, but it 

### 11 seems to be that our take on this in distilling a lot of the 

### 12 other discussions, that level of concern as derived at kind of 

### 13 lower tier more abductionistic levels are a good empirical way 

### 14 to arrive at those kind of baseline levels that need to be 

### 15 vetted and verified at upper levels. Those are obviously a very 

### 16 good starting point. 

### 17 But because of the transient property from one 

### 18 level to another is not 100 percent obviously, that I think that 

### 19 that would provide a good starting point, but it's not 

### 20 necessarily going to be exactly the same as one increases in 

### 21 biological complexity and in environmental variability. So, it 

### 22 seems an appropriate starting point, but that it should be 

### 23 modulated depending on the different tiers in which they're 

### 24 being tested. 

### 25 **DR. DANIEL SCHLENK:**Okay. Dr. Berenbaum? 

### 26 **DR. MAY BERENBAUM:**I just like to repeat the

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1 call for input from our toxicologist colleagues who have 

### 2 examined these kinds of estimates in much broader context than 

### 3 we have. 

### 4 **DR. DANIEL SCHLENK:**Dr. Pettis, let me guess 

### 5 what you're going to say. 

### 6 **DR. JEFF PETTIS:**I'm gonna say that our level 

### 7 of concern among the three of us was raised that we were looking 

### 8 for help basically. 

### 9 **DR. DANIEL SCHLENK:**Okay. With that, anybody 

### 10 want to jump in? Dr. Klaine, not saying any names. 

### 11 **DR. STEPHEN KLAINE:**You know, this thing is 

### 12 really confounded by the fact that as you move from one level to 

### 13 another. You've got this issue of single organisms moving to 

### 14 more complex systems. And I'm really not sure if we're able to 

### 15 quantify the level of concern that would be the most useful as 

### 16 you move into these higher tiers. 

### 17 **DR. THOMAS STEEGER:**Could I make a point of 

### 18 clarification? 

### 19 **DR. DANIEL SCHLENK:**Sure, Dr. Steeger. 

### 20 **DR. THOMAS STEEGER:**The level of concern is 

### 21 applied to the Tier I laboratory-based studies only. The higher 

### 22 tiers studies that are at the colony level do not have a level 

### 23 of concern that's applied to them. 

### 24 **DR. DANIEL SCHLENK:**Thanks for that 

### 25 clarification. This is just for the Tier I. 

### 26 **DR. STEPHEN KLAINE:**All right. Okay. I

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1 misread the question. 

### 2 **DR. DANIEL SCHLENK:**Are you done? No? Are 

### 3 you thinking? 

### 4 **DR. STEPHEN KLAINE:**For the moment. 

### 5 **DR. DANIEL SCHLENK:**Okay. Dr. Hunt? 

### 6 **DR. GREG HUNT:**Well, my understanding is this 

### 7 LOC 0.4 corresponds to about 10 percent mortality. Given that a 

### 8 lot of the suggestions are to follow the larvae to pupation and 

### 9 to have longer observation of adults in survival analysis, I 

### 10 don't see how this particular number applies. But probably some 

### 11 number could be developed or some qualitative, you know, in 

### 12 terms of development maybe, there could be a number developed 

### 13 there as well. But, anything that is enough standard deviations 

### 14 from the norms. 

### 15 **DR. DANIEL SCHLENK:**Mr. Pistorius? 

### 16 **MR. JENS PISTORIUS:**With limitations of not 

### 17 knowing what exactly which substances, which products exactly 

### 18 were behind the data, that table that Dr. Steeger showed the 

### 19 other day on how many substances would actually pass the first 

### 20 trigger. Given that there is a need in my opinion for tiered 

### 21 approach in risk assessment and also keeping into account that 

### 22 we might miss a very small number, I think really a very small 

### 23 number of active with such a tiered approach, I have the 

### 24 impression that this level of concern is a conservative measure 

### 25 and an appropriate screen considering the comparison with the HQ 

### 26 data, which has been for a long time used in Europe and very

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1 well information we have on incidents in different countries. 

### 2 The clearly improved conservatism compared to this approach, I 

### 3 don't remember which presentation it was but, in one 

### 4 presentation it was said that this approach is about a factor 

### 5 3.6 more conservative than the approach that we had. So that 

### 6 might be a German interpersonal perspective. 

### 7 **DR. DANIEL SCHLENK:**Thanks. Dr. Klaine? 

### 8 **DR. STEPHEN KLAINE:**Yeah, I agree. It's very 

### 9 conservative. My hesitation before really comes from trying to 

### 10 get my head around the fact that when we do this with other 

### 11 species, when you have an acute number, you get some reflection 

### 12 about the impact on the population. In this case, we did it 

### 13 with single organisms, and we really don't know how that 

### 14 reflects that actual health of the hive or the ultimate 

### 15 population of bees. So, I agree it's conservative. I'm 

### 16 actually bordering on the fact that it might be too 

### 17 conservative. I'm not sure how to quantify that. 

### 18 **DR. DANIEL SCHLENK:**Anyone else want to weigh 

### 19 in on that? For what it's worth, I think it's conservative in 

### 20 terms of you know, what it's trying to do and the function that 

### 21 it's trying to serve in a Tier I screen that seems to have very 

### 22 limited error with regard to false negatives, which is what 

### 23 you're trying to do, right? Yes? 

### 24 **DR. STEPHEN KLAINE:**Yeah, and where I 

### 25 hesitate is what is the right number. 

### 26 **DR. DANIEL SCHLENK:**Isn't it 42? Yes? Dr.

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1 Pettis? 

### 2 **DR. JEFF PETTIS:**I guess just a question to 

### 3 EPA and Dr. Hunt touched on this. This value roughly relates to 

### 4 10 percent mortality, and that would be above whatever the 

### 5 control mortality is. So, if you had control mortality of 10 

### 6 percent, then you would have to see 20 percent mortality in 

### 7 another group. Is that correct? 

### 8 **DR. THOMAS SEEGER:**This is Tom Seeger. 

### 9 Essentially it's taking that LD50 value. The slope that is 

### 10 calculated off of all of the acute toxicity data that we have 

### 11 for oral and contact toxicity and estimating what would the 

### 12 value have to be to back it down to control mortality. So, 

### 13 you're essentially bringing it down to a NOAEC that would be - 

### 14 well you're not able to distinguish between the treated and the 

### 15 control value. So, the chronic LOC is set to 1, because the 

### 16 NOAEC is compared to the exposure value and the acute value 

### 17 because we're backing that LD50 back down to what we think is 

### 18 background mortality, acute mortality is set to 10 percent. 

### 19 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 20 **DR. MAY BERENBAUM:**Point of clarification 

### 21 that you say all available acute mortality data, toxicity data, 

### 22 are those from forager or workers, right? Are there any larval 

### 23 data that is factored in? 

### 24 **DR. THOMAS STEEGER:**These data were based on 

### 25 adult - young worker data, that's correct. 

### 26 **DR. DANIEL SCHLENK:**So, do we have a

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1 consensus that 0.4 is valid? That's what the question is 

### 2 asking, right? Yes? If you don't then you need to come up with 

### 3 another number, I guess. That's the question. Yeah, Dr. 

### 4 Klaine? 

### 5 **DR. STEPHEN KLAINE:**Steve Klaine. You know, 

### 6 the goal is to be conservative and to not make errors. So, you 

### 7 know if we're gonna go in that direction, then this is in fact 

### 8 conservative. I think that as more data become available in 

### 9 terms of the relationship between individuals and hive health, I 

### 10 think that you may want to hold out the opportunity to change 

### 11 that. 

### 12 **DR. DANIEL SCHLENK:**Yeah, what I would say 

### 13 too is particularly if you're going to change the assay to the 

### 14 other assays that have been discussed, that number may actually 

### 15 change as well if we go with some of the alternative assays that 

### 16 have been proposed, at least durations. Yeah, Dr. Fefferman? 

### 17 **DR. NINA FEFFERMAN:**So, this is actually a 

### 18 strange context for me to propose this, but this is actually one 

### 19 of the context in which I would propose a sensitivity analysis 

### 20 from the end, not of the risk assessment, but for the risk 

### 21 manager. Because the only danger I see in being too 

### 22 conservative is that if you accidentally make it so conservative 

### 23 that almost every viable benefit already puts you over the 

### 24 threshold, then it makes it a yes or a no, do we want the 

### 25 benefit at all question. 

### 26 So, if there is a way to do a sensitivity on

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1 the relative benefits from the management perspective in 

### 2 interpreting the outcome of the risk assessment based on the 

### 3 sensitivity of the cutoff for how conservative that should be, 

### 4 there is a utility of not being so conservative that the answer 

### 5 is anything you do is equally bad, so maximize your gains if 

### 6 you're going to do anything because that can push - am I being 

### 7 clear enough? Okay. Cool. 

### 8 **DR. DANIEL SCHLENK:**Did you get that Dr. 

### 9 Tarpy? Any questions of clarification? Yeah, we're good? 

### 10 Okay. We're going to go forward to letter C then. 

### 11 **DR. THOMAS STEEGER:**This is Tom Steeger, 

### 12 question 13C - please comment of the quantitative aspect of the 

### 13 screening-level Tier I assessment and the use of Tier II and 

### 14 Tier III whole-hive studies to qualitatively characterize risk. 

### 15 **DR. DANIEL SCHLENK:**Dr. Tarpy? 

### 16 **Dr. DAVID TARPY:**This is Dave Tarpy. I think 

### 17 our comment on the answer to this one is because it is very 

### 18 similar to the others about extrapolating from one level to the 

### 19 others is a little more difficult in light of the social life 

### 20 history of honey bees. So, this pretty much the same caveat is 

### 21 given as to the previous question. 

### 22 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 23 **DR. MAY BERENBAUM:**I concur. 

### 24 **DR. DANIEL SCHLENK:**Dr. Pettis? 

### 25 **DR. JEFF PETTIS:**I concur. 

### 26 **DR. DANIEL SCHLENK:**Any other panel members?

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1 So what it sounds like is this has been addressed in earlier 

### 2 questions. Are you guys okay with that? Yeah? Okay. Letter 

### 3 D. 

### 4 **DR. THOMAS STEEGER:**This is Tom Steeger, 

### 5 question 13D - please comment on the assumption that the 

### 6 effects on individual bees measured in laboratory studies must 

### 7 be considered in the context of whole colony studies conducted 

### 8 under semi-field and full-field conditions. 

### 9 **DR. DANIEL SCHLENK:**Dr. Tarpy. 

### 10 **DR. DAVID TARPY:**This is Dave Tarpy. Again, 

### 11 as has been covered in many of the questions. I think it has 

### 12 been pretty well flushed out, but the assumption that 

### 13 individuals serve as a valuable proxy for the whole group is 

### 14 pretty much nullified and doesn't exist, which is why we spend 

### 15 so much time emphasizing alternative means of testing groups in 

### 16 vitro and in vivo to try to more realistically approximate in 

### 17 vivo colony dynamics. 

### 18 So, there should be real degrees of care to 

### 19 make broad-sweeping inferences of data on at the individual 

### 20 honey bee level to full-field colony phenotypes. So, by being 

### 21 able to assess at Tier I with social groups and kind of more 

### 22 biologically relevant bioassays that will go a long way to 

### 23 alleviate this particular question. 

### 24 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 25 **DR. MAY BERENBAUM:**I concur. 

### 26 **DR. DANIEL SCHLENK:**Dr. Pettis?

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1 **DR. JEFF PETTIS:**I have just a bit to add. 

### 2 Just in looking at effects that may be realized in Tier I, they 

### 3 may help guide you and then what you might look for in full 

### 4 colony or semi-field or field studies, but there is a caution. 

### 5 I think Dave has already touched on it that an effect that you 

### 6 might see in a cage with individual bees, there are so many 

### 7 compensation mechanism of grooming and prophylaxis and things 

### 8 that happy at the colony level. 

### 9 That effect may totally disappear and you may 

### 10 be looking for other effects that may happen at the colony 

### 11 level. Food transfer may magnify affects towards the queen, 

### 12 things like that. So, I think it's important to look at the 

### 13 effect that Tier I and move forward. But the social dynamics 

### 14 and colony interactions can really - it's really hard to predict 

### 15 how those individual measures might be realized in the colony. 

### 16 So again, kind of reiterating what is being said. 

### 17 **DR. DANIEL SCHLENK:**Other panel members? 

### 18 Okay. We go back to the Agency. Any questions of 

### 19 clarification? 

### 20 **DR. THOMAS STEEGER:**Point of clarification. 

### 21 In the absence of those other socially proper studies, the 

### 22 current guidelines that exist serving as the basis for Tier I 

### 23 studies and using them in a screen, does the Panel concur or can 

### 24 you comment on the use of the currently available guidelines. 

### 25 **DR. DANIEL SCHLENK:**Dr. Tarpy? 

### 26 **DR. DAVID TARPY:**So, this is Dave Tarpy. I

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1 guess that they are informative, but not as informative as they 

### 2 could be because of those complexities. So I think, and the 

### 3 Panel can agree or disagree, but that's why we've spent so much 

### 4 time urging the development of more biologically relevant in 

### 5 vitro bioassays that would kind of harness that a bit better. I 

### 6 mean obviously, some informed information is better than no 

### 7 information, but I think that because of this disconnect of the 

### 8 individual versus group level that those data derive at the 

### 9 individual level ought to be interpreted with much greater 

### 10 caution. 

### 11 **DR. DANIEL SCHLENK:**Dr. Berenbaum? 

### 12 **DR. MAY BERENBAUM:**One additional benefit of 

### 13 using the existing studies as approaches is that it would allow 

### 14 harmonization of internation efforts and an acquisition of a 

### 15 greater data base to use in trying to make that connection. But 

### 16 I just want to echo the call for reexamining assay design to be 

### 17 more reflective of biological reality. 

### 18 **DR. DANIEL SCHLENK:**Dr. Ostiguy? 

### 19 **DR. NANCY OSTIGUY:**I want to actually 

### 20 strongly support the use of Tier I data this point, because we 

### 21 don't have anything else to work with. It is much better than 

### 22 nothing. Actually, so much better than nothing that it's almost 

### 23 no comparison, which doesn't negate the fact that there might be 

### 24 better ways to assess things. I do think that they will provide 

### 25 us with a sufficiently sensitive set of criteria for making 

### 26 decisions that we should not commit too many type II errors.

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1 **DR. DANIEL SCHLENK:**Okay. Mr. Pistorius? 

### 2 **MR. PISTORIUS:**Yes, I think that the current 

### 3 guidelines are very suitable. We know that there are a few 

### 4 guidelines or there is especially one guideline, which has to be 

### 5 further developed or established at all at the Tier I level. 

### 6 This is yet to be done. I would say as a general remark that 

### 7 some of the existing guidelines, also the OECD 75 and EPPO 170 

### 8 need a little bit of updating. But I think those were the 

### 9 points that were discussed in our discussions here already, and 

### 10 they are minor points, which do not criticize the whole approach 

### 11 that is mentioned in the guidelines. 

### 12 **DR. DANIEL SCHLENK:**Dr. James, did you have 

### 13 you hand up earlier? No? Yes? 

### 14 **DR. ROSALIND JAMES:**I don't like the current 

### 15 guidelines. I think they're - we're having problems with bees. 

### 16 They are not a very good assessment of what our effects are 

### 17 going to be. So, I guess I don't know that they are much better 

### 18 than nothing. We're talking about the 48 hour contact toxicity 

### 19 on adult tests, right? When you say current guidelines? At 

### 20 Tier I. They're recommended in the White Paper or the current 

### 21 guidelines, I'm not sure what you're asking I guess. 

### 22 **DR. THOMAS STEEGER:**This is Tom Steeger, 

### 23 point of clarification. Two points of clarification. We are 

### 24 talking about now actually quantifying risk using a formal 

### 25 process, which has not been done before. We are also talking 

### 26 about modifications to the testing process that will give us a

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1 much better understanding of not just adult toxicity, but larval 

### 2 toxicity and a path forward to where there are potential risks 

### 3 that are identified using these screening level tests to 

### 4 transitioning the higher tiered tests so would better enable us 

### 5 to understand whole colony effects. 

### 6 **DR. ROSALIND JAMES:**This is Rosalind. And 

### 7 when you said current guidelines, that's what you're referring 

### 8 to? 

### 9 **DR. THOMAS STEEGER:**This is tom Steeger. I'm 

### 10 discussing what has been laid out in the White Paper. 

### 11 **DR. ROSALIND JAMES:**Okay. 

### 12 **DR. DANIEL SCHLENK:**Okay. Dr. Fefferman? 

### 13 **DR. NINA FEFFERMAN:**So I think the Tier I 

### 14 studies for individual level are really good for the individual 

### 15 effects. Where I find the disconnect for the context of the 

### 16 question of scaling those up is actually propagates back down, 

### 17 and I don't remember seeing quantitative ways to characterize 

### 18 the sublethal effects that sufficiently trigger Tier I response 

### 19 to go to Tier II. And I don't know if that's because I'm just 

### 20 not remembering it or because it's not there. I know there was 

### 21 discussion of how to try to measure some sublethal effects, but 

### 22 I don't remember seeing how that gets interpreted in the 

### 23 conceptual flow model for then going from okay, we've finished 

### 24 our Tier I study, we saw the sublethal effects. That means we 

### 25 now really do need to go to Tier II. That's the part where I 

### 26 start not knowing how to get my mind around if this working

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1 well. 

### 2 **DR. DANIEL SCHLENK:**Any other comments? Dr. 

### 3 Hunt? 

### 4 **DR. GREG HUNT:**I think the guidelines 

### 5 proposed in the White Paper are a good advance - a good 

### 6 improvement. But I don't think there was proposed the use of 

### 7 sublethal effects. In Tier I, it was proposed as something that 

### 8 might be useful in the future. I think that what the panel is 

### 9 saying is that there could be improvements in sublethal effects 

### 10 included for example, following the growth of the larva to 

### 11 pupation and looking at biological endpoints and things of that 

### 12 sort. So I think that what's proposed, if I'm understanding 

### 13 correctly is good, but we're suggesting there could be some easy 

### 14 improvements. Maybe not so easy. 

### 15 **DR. DANIEL SCHLENK:**Any other comments? Dr. 

### 16 Klaine? 

### 17 **DR. STEPHEN KLAINE:**Yeah, I just want to 

### 18 weigh in on this. From the panel discussions and all the 

### 19 discussions we've had, it just at that level, it appears that 

### 20 mortality is just the wrong endpoint and it's difficult to 

### 21 interpret to move forward with that. I think that's why 

### 22 everyone is calling for sublethal endpoints. So that the issue 

### 23 with behavioral endpoint or other endpoints as they are 

### 24 developed would better able this movement from single organisms 

### 25 in Tier I up to the semi-field and field studies. 

### 26 **DR. THOMAS STEEGER:**Point of clarification?

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1 **DR. DANIEL SCHLENK:**Sure. 

### 2 **DR. THOMAS STEEGER:**For the - in the White 

### 3 Paper, it describes that the sublethal effects at this time 

### 4 because of the absence of appropriate linkages to assessment 

### 5 endpoints, while they would not be used quantitatively, they 

### 6 would be used qualitatively and that they would be considered 

### 7 with the other lines of evidence available to determined whether 

### 8 higher tier testing would be required. So, the fact that the 

### 9 current battery of Tier I studies tend to focus on mortality as 

### 10 the primary effect. There is the opportunity that additional 

### 11 endpoints may be reported, but we're not confined to only 

### 12 considering the mortality endpoint. We do leave open the option 

### 13 to qualitatively consider whether the sublethal effects are 

### 14 sufficiently compelling to warrant with other lines of evidence 

### 15 transitioning to higher tier testing. 

### 16 **DR. DANIEL SCHLENK:**I believe that is going 

### 17 to be the next sub-question, right? Yes. Dr. Bradbury, did you 

### 18 have something you want to say? No, he just got - okay. Great. 

### 19 Well with that, are you okay with the answer that we received 

### 20 for this sub-question? So, let's call it quits for now. I'm 

### 21 sensing a little bit of brain drain here this afternoon. So, 

### 22 we'll start in the morning with this. There are three more 

### 23 subsections yet, and we'll start in the morning. And to get 

### 24 better conversation, we'll start in the morning on sub-question 

### 25 E and start with the weight of evidence at that point. Okay. 

### 26 Everybody will come back fresh in the morning. Okay. So with

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1 that, do you need to say anything Fred. Okay. We're adjourned 

### 2 for today. We will meet a nine o'clock tomorrow morning. 

### 3 (WHEREUPON the meeting was adjourned for the 

### 4 day) 

### 5 **DR. DANIEL SCHLENK:**As with yesterday, I 

### 6 think we will skip the introductions and get into our charge 

### 7 questions. 

### 8 Before we move forward I believe the Agency 

### 9 had a couple of comments they wanted to make before we move into 

### 10 the next letter for the charge questions. 

### 11 **DR. THOMAS STEEGER:**Thank you very much Dr. 

### 12 Schlenk. This is Tom Steeger with Environmental Fate and 

### 13 Effects Division. 

### 14 Just as a point of clarification in the 

### 15 discussions as we are talking about higher tier studies and how 

### 16 they relate to the laboratory-based studies. The intent of 

### 17 those higher tier studies as described in the White Paper is to 

### 18 focus in on particular uncertainties that have been identified 

### 19 in either the lower tier studies or incident reports that we 

### 20 might have received or open literature studies, so that you are 

### 21 addressing very specific risk hypotheses and not generalized 

### 22 questions. 

### 23 Our experience in the agency with other taxa 

### 24 and certainly our experience with some of the pollinator studies 

### 25 that have been submitted in the past, is that they are 

### 26 attempting to answer too many questions and in the process you

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1 do not end up with any answers. So please keep in mind that the 

### 2 intent of higher tier studies is to focus in on particular 

### 3 hypotheses. 

### 4 **DR. DANIEL SCHLENK:**Dr. Potter, I think you 

### 5 had a paper you found for your comments yesterday? 

### 6 **DR. THOMAS POTTER:**Yes, I did. This was in 

### 7 regards to Question 7, and there was a question from the EPA 

### 8 staff about examining dust emissions and what might be a 

### 9 reasonable approach to that. 

### 10 I did some digging last night and I found an 

### 11 article that was published in Environmental Science and 

### 12 Technology in January of this year, by an Italian research 

### 13 group. I'll just enter that into the record. 

### 14 It provides some emission factors for some of 

### 15 the seed coating insecticide, and using the emission factors and 

### 16 some of the subsequent deposition data that they were able to 

### 17 obtain we should be able to come up with an exposure estimate 

### 18 and see how that compares to the value that has already been 

### 19 used in the White Paper. I'll make sure we get that into the 

### 20 record. 

### 21 **DR. DANIEL SCHLENK:**Mr. Pistorius. 

### 22 **MR. JENS PISTORIUS:**Thank you. May I add on 

### 23 this that is also the report of the opponent, whether reported 

### 24 on a wider scale on issues on the dust and this paper is one of 

### 25 the issues that is also in this report. 

### 26 And all of this and even more data is also

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1 mentioned in this report, which we include in the document. 

### 2 **DR. DANIEL SCHLENK:**Dr. Berenbaum. 

### 3 **DR. MAY BERENBAUM:**Is this the point at which 

### 4 I can introduce.? No, okay. 

### 5 **DR. DANIEL SCHLENK:**Okay, we left off last 

### 6 time at Charge Question 13d, so who wants to read in Charge 

### 7 Question 13e? 

### 8 **MR. KEITH SAPPINGTON:**Please comment on the 

### 9 proposed use of a weight-of-evidence approach based on 

### 10 information obtained from multiple tiers of risk assessment for 

### 11 characterizing pesticide risks to honey bees. 

### 12 **DR. DANIEL SCHLENK:**The lead on that is Dr. 

### 13 Tarpy. 

### 14 **DR. DAVID TARPY:**Good morning everybody. The 

### 15 discussants on this particular point just again want to 

### 16 reiterate that it is always difficult to move from one level of 

### 17 biological organization to others but we definitely concur and 

### 18 agree with the weightof-evidence approach by trying to 

### 19 synthesize the information from these multiple tiers. 

### 20 And so we also thought that high throughput 

### 21 methods that are reliable and repeatable are obviously going to 

### 22 be the most valuable and hopefully this discussion from this 

### 23 week will be able to generate ideas of how to do those types of 

### 24 screening techniques that account for the biological complexity 

### 25 that is added by social organisms such as honey bees. 

### 26 But then we also believe that because the

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1 basil unit of selection for a social insect like honey bees is 

### 2 the colony level, we feel that there should be a centralized 

### 3 intendancy of the weight-of-evidence towards the Tier II level 

### 4 that is going to be the most informative. This is also going to 

### 5 be discussed in the next sub-question for Charge Question 13. 

### 6 **DR. MAY BERENBAUM:**Just two additional 

### 7 comments that we discussed. One is that certain parameters may 

### 8 be very readily quantifiable and statistically analyzed but they 

### 9 may not necessarily be as informative as less easily measured 

### 10 criteria, which is an argument for weight-of-evidence. 

### 11 And in addition the consideration that the 

### 12 impact that is being assessed is on a workforce rather than 

### 13 simply an organism, it is not just an up or down, alive or dead 

### 14 criterion, and it is quality as well. 

### 15 So quality issues are not as readily 

### 16 quantifiable so weight-of-evidence of multiple indicators would 

### 17 be particularly appropriate for assessing honey bee pesticide 

### 18 sensitivity. 

### 19 **DR. JEFF PETTIS:**Just one additional comment 

### 20 along the same lines. 

### 21 We talked about biological effects and 

### 22 statistically significant effects and this weight-of-evidence 

### 23 approach will allow us to consider those biological effects 

### 24 that, because of sample size in Tier II and Tier III may be 

### 25 reduced and we cannot reach statistical significance, we should 

### 26 keep in mind there is biological effect and I think this

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1 weight-of-evidence approach allows us to do that. 

### 2 **DR. DANIEL SCHLENK:**Other panel members. 

### 3 Really, wow. Great. 

### 4 Let me go back to Mr. Sappington, do you have 

### 5 any questions or clarifications? Okay, great. Let's go ahead 

### 6 and read in Charge Question 13f. 

### 7 **MR. KEITH SAPPINGTON:**Charge Question 13f; 

### 8 please comment on how best to characterize overall uncertainty 

### 9 or weigh different areas of uncertainty in risk 

### 10 characterization. 

### 11 **DR. DAVID TARPY:**So this again is what I 

### 12 alluded to in sub-question e that Tier III studies we feel are 

### 13 much more prone to type II errors and false negatives and 

### 14 therefore, Tier II studies seem to be much more robust in trying 

### 15 to reduce uncertainty in this type of risk characterization. 

### 16 And that is because type II studies are more prone to type I 

### 17 errors but therefore are much more conservative. 

### 18 Tier I studies, of course, are very robust and 

### 19 repeatable but at the individual level do not translate as well 

### 20 to whole colony systems, and so the weight-of-evidence at that 

### 21 screening level, there ought to be caution in interpretation of 

### 22 that. 

### 23 So again, it places the real emphasis of 

### 24 testing and screening at the Tier II level at whole colony in 

### 25 vivo type settings. 

### 26 **DR. MAY BERENBAUM:**I concur.

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1 **DR. JEFF PETTIS:**I concur. 

### 2 **DR. DANIEL SCHLENK:**Other panel members. Dr. 

### 3 Fefferman. 

### 4 **DR. NINA FEFFERMAN:**Good morning. Just a 

### 5 brief comment on it; I concur with everything that has been 

### 6 said. I want to try and disambiguate. In risk assessment, 

### 7 especially in modeling right there are. I'm sorry. 

### 8 **UNKNOWN SPEAKER:**Disambiguate? 

### 9 **DR. NINA FEFFERMAN:**Yes, to choose a part. I 

### 10 do not actually mean a clarification of something that has 

### 11 already been said but I mean it using a part of the dual 

### 12 meanings of the word uncertainty. Did I invent a word? 

### 13 Uncertainty itself has two very meaningful and 

### 14 very very very functionally different sources. One is 

### 15 biological stochasticity leading to different ranges of 

### 16 measurements that are accurate in their measurement and the 

### 17 other is not knowing what those values would be. 

### 18 And so treating those as one thing can lead 

### 19 itself to some confusion. I just wanted to make sure that that 

### 20 was somewhere in the conversation. 

### 21 **DR. DANIEL SCHLENK:**Other comments? 

### 22 **MR. JENS PISTORIUS:**When we address 

### 23 uncertainty, for instance with the investigation of incidence, 

### 24 that may be used to describe the risk that has occurred under 

### 25 practical conditions but nevertheless it only has a limited use 

### 26 for actually the risk assessment.

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1 If you cannot prove that because you have the 

### 2 proper use and you have the improper use. I guess it is the 

### 3 same, yes like in Europe to assess as a risk assesses the proper 

### 4 use of the substance. This is basically one uncertainty that 

### 5 you have to take into account when considering incidents. 

### 6 I think basically you have on all different 

### 7 tiers uncertainties starting with lower uncertainties from Tier 

### 8 I; they increase a little bit, Tier II. I think Tier II studies 

### 9 with worst case exposure have quite a low uncertainty. 

### 10 Depending on the number and the design of the 

### 11 studies you may have a small uncertainty or medium uncertainty 

### 12 also in the tier test, if the study design is bad you may have a 

### 13 very high uncertainty for sure. But it depends on also the 

### 14 number of studies that are actually handed in. 

### 15 So I think you have to take those into account 

### 16 but nevertheless even when you have a certain number of 

### 17 incidents does not have to change the outcome of the risk 

### 18 assessment necessarily, if it is not proven or if you cannot 

### 19 link it to a proper use. 

### 20 For instance, we get a small number of 

### 21 poisoning incidents with a few insecticidal substances, spray 

### 22 application, every year. But then again in most cases where we 

### 23 could trace it down - we cannot trace it down in all cases - we 

### 24 actually found out that it was improper uses. 

### 25 So too high amount applied, wrong crop applied 

### 26 mitigation measures not used. But on the other hand, especially

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1 the incident may be used to give a feedback on how good is 

### 2 actually the risk assessment to be incorporated in the real life 

### 3 situation. 

### 4 This is a part which also may give a feedback 

### 5 on the uncertainty that you create in your risk assessment 

### 6 process. And then in some cases may even be valid enough to 

### 7 say, Okay this application is again a risk management issue not 

### 8 practical for the farmers to achieve. 

### 9 **DR. DANIEL SCHLENK:**Any other comments? Yes, 

### 10 Dr. Ostiguy. 

### 11 **DR. NANCY OSTIGUY:**I'm not sure if I 

### 12 understood Jens' last part well enough. And I do not know if 

### 13 EPA can address improper use, but when you have an overwhelming 

### 14 - and this of course is a judgment call - a large number of 

### 15 improper uses, I do think that that has to weigh into whether or 

### 16 not we are going to use a particular substance. 

### 17 **DR. DANIEL SCHLENK:**Other comments? Mr. 

### 18 Pistorius. 

### 19 **MR. JENS PISTORIUS:**I think that's basically 

### 20 then the part of the risk management to see how good it is 

### 21 actually possible to incorporate this behavior of normal people 

### 22 into this and it is not enforcement action but again looking 

### 23 from at all states we have got different responsibilities. 

### 24 And so it is not a risk assessment; it is a 

### 25 risk management issue and in all cases it is not the federal but 

### 26 then it is a state issue of ensuring that those enforcements are

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1 done; for clarification of my statement. 

### 2 **DR. DANIEL SCHLENK:**Any other comments for 

### 3 this particular question? Mr. Sappington, any questions or 

### 4 clarifications? Dr. Steeger. 

### 5 **DR. THOMAS STEEGER:**Just as a point of 

### 6 clarification on the incident data. We do make an effort to 

### 7 distinguish where chemicals have been used properly or misused, 

### 8 and actually a trend in the misuse of a chemical can inform risk 

### 9 management decisions as well; that can be a problem in and of 

### 10 itself. Thank you. 

### 11 **DR. DANIEL SCHLENK:**All right, let's go ahead 

### 12 and move on to our last sub-question for 13. 

### 13 **MR. KEITH SAPPINGTON:**Charge Question 13g, 

### 14 please comment on how to focus or prioritize uncertainties when 

### 15 designing and interpreting Tier II and Tier III studies. 

### 16 **DR. DANIEL SCHLENK:**Dr. Tarpy. 

### 17 **DR. DAVID TARPY:**So most of the 

### 18 recommendations and issues that have been brought up that weigh 

### 19 in on this particular question, we feel have been discussed 

### 20 previously so we do not really want to recapitulate them here. 

### 21 But things like ensuring adequate and 

### 22 sufficient sample size to increase statically power and then 

### 23 weighing that evidence to focus uncertainties is good. 

### 24 Replication of study obviously, and focusing at Tier II levels, 

### 25 at the colony level for testing phenotypic effects, are all 

### 26 means of really focusing the designs of those types of studies

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1 that seems to be a priority. 

### 2 Also, I understand the real need of wanting to 

### 3 focus on one or two critical measures and hypotheses, but we 

### 4 have been discussing all week the complexities of different 

### 5 routes of exposure and different modes of action and different 

### 6 ill effects, sometimes very subtle on social insect colonies 

### 7 that can all result in ill health. 

### 8 It is hard to always identify kind of with one 

### 9 broad stroke that this is the one thing that needs to be 

### 10 measured and if that is okay then everything else is going to be 

### 11 okay with honey bee colonies. So I think including those 

### 12 complexities in these knowing that one size fits all is not 

### 13 going to be the case in all of these Tier II and Tier III 

### 14 studies is definitely something to keep in mind. 

### 15 But hopefully, as we will discuss in Charge 

### 16 Question 14 that there may be ways to try and narrow those down 

### 17 and identify some of the real key aspects that can be focused 

### 18 on. 

### 19 **DR. MAY BERENBAUM:**I just want to add one 

### 20 other factor to address uncertainties in addition to the 

### 21 geographic replication, season replication. I like us to keep 

### 22 in mind genetic variability within managed populations of Apis 

### 23 mellifera. 

### 24 **DR. JEFF PETTIS:**I concur with what have been 

### 25 said. 

### 26 **DR. DANIEL SCHLENK:**Other panel members? Dr.

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1 Pistorius. 

### 2 **MR. JENS PISTORIUS:**When we focus on how to 

### 3 prioritize uncertainties when designing and later on 

### 4 interpreting tier studies, when designing I think that you have 

### 5 to have a look at the concern that you get from the lower tiers. 

### 6 You have to identify the relevant root of 

### 7 exposure that is of concern to you. For instance, you get 

### 8 different toxicity for certain insecticide; you may get 

### 9 substances with a high or/and low contact toxicity and vice 

### 10 versa, or substances which have both high levels. 

### 11 Such information from the lower tier basically 

### 12 is needed and should be written down what the concern is and 

### 13 then used to be designing and focusing on the design for the 

### 14 Tier II and Tier III studies. For instance, also what I 

### 15 mentioned with the sub-lethal behavioral effect, if you have 

### 16 such a study where you have severe sub-lethal behavioral effect 

### 17 then you might have to change the design accordingly and maybe 

### 18 focus differently on the different aspects that you can test 

### 19 with your Tier II and Tier III studies. 

### 20 What we have not mentioned in this round is 

### 21 basically, "What are the different options that you can test in 

### 22 such a Tier III study." 

### 23 We just talked about tier studies and this is 

### 24 basically standard tier study, but there are additional methods 

### 25 available and we cannot elaborate on them all in this short 

### 26 time, which would actually be able to address specific concerns.

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1 We have shortly mentioned, for instance, the 

### 2 impact on homing behavior. There are tests available which may 

### 3 be used to address such questions. 

### 4 In a standard field test, according to EPPO, 

### 5 for instance, you do a brood assessment according to the leave 

### 6 or failure method, you look at all the brood and the number of 

### 7 brood and this is basic in estimation. But you can additionally 

### 8 combine it with the OECD 75 method where you actually do mapping 

### 9 of the brood and follow the individual brood. 

### 10 So I think this can just be a general comment, 

### 11 but I think the level of the concern that you get from the tier 

### 12 I study is basically the most important one which help you for 

### 13 designing this. 

### 14 **DR. DANIEL SCHLENK:**Dr. James. 

### 15 **DR. ROSALIND JAMES:**Jens that was an 

### 16 excellent answer I think. My point seems minor in comparison. 

### 17 I would add to your answer, I think yesterday 

### 18 some comments were made to me by the EPA folks that when we had 

### 19 discussions about the Tier I experimental design, that we were 

### 20 throwing out sort of too many measures that could be taken. And 

### 21 that is one way that they could focus down to which of these 

### 22 measures are the ones that are most important and base it on the 

### 23 results of the Tier I and the Tier II effects that were seen. 

### 24 The other thing I was going to add about 

### 25 helping with designing an uncertainty is proper replication and 

### 26 controls. Honey bees are highly variable systems and then when

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1 you go to a colony level you really need to make sure that the 

### 2 experiments are replicated enough and that the controls are good 

### 3 controls. 

### 4 **DR. JEFF PETTIS:**Just building on those 

### 5 comments; another good example is if you had a concern raised in 

### 6 Tier I that maybe there was chronic exposure that was going to 

### 7 shorten the longevity of adult worker bees and one ramification 

### 8 for that would be at the colony level maybe overwintering 

### 9 success. 

### 10 So designing a study to get at where the 

### 11 colonies could adequately overwinter after exposure, that kind 

### 12 of thing. And those get longer in duration, they are harder to 

### 13 control, but they are possible. 

### 14 **DR. GREG HUNT:**Along those same lines, if it 

### 15 is going to affect individual longevity in Tier II and Tier III, 

### 16 it is pretty simple to emerge some bees in an incubator, put 

### 17 those newly emerged bees that are paint-marked in the colony and 

### 18 then census them over time to get an average longevity. 

### 19 **MR. JENS PISTORIUS:**To add to what Jeff 

### 20 Pettis has said, I think it's completely right. 

### 21 One idea that might help you in designing, or 

### 22 looking at when do I have to do an overwintering or long-term 

### 23 study. This, of course, is basically self explaining, but use 

### 24 for instance the substance properties; as the persistence and 

### 25 the degradation and also the mode of application if it is an 

### 26 acute spray treatment. Also taking into account the information

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1 that is available on other organisms or on the efficacy side, 

### 2 the mode of action that is proposed. 

### 3 **DR. MARTHA SANDY:**I have a question for the 

### 4 folks here that have the expertise. You are saying the honey bee 

### 5 colonies are very variable; I wonder how would you design a 

### 6 study to take that into account? Would you have sister queens 

### 7 in the different colonies or what would be the ways to do that? 

### 8 **DR. NANCY OSTIGUY:**There are some things you 

### 9 can do to reduce that issue. You would pick the same line, so 

### 10 it is all Italians or all carniolans. You would pick sister 

### 11 queens. 

### 12 As much as possible you would have the 

### 13 colonies in a similar micro-environment. I'm sure others have 

### 14 suggestions. 

### 15 **DR. GREG HUNT:**What she said, but you would 

### 16 have sister queens all mated in the same mating yard. Of 

### 17 course, they are mating to about a dozen different males so you 

### 18 have a lot of genetic diversity in the colony but hopefully that 

### 19 all averages out and then you have of course more replication. 

### 20 **DR. NINA FEFFERMAN:**But that all said, you 

### 21 need the replication of those control samples in order to make 

### 22 sure that you are not looking just within one line of effect. 

### 23 **DR. STEPHEN KLAINE:**I just wanted to ask, do 

### 24 we even know enough about the variation in order to do a power 

### 25 analysis; so we even have an idea of how many replicates we need 

### 26 in these designs?

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1 **DR. GREG HUNT:**The answer is probably not 

### 2 because it depends on which study you are doing and some studies 

### 3 are going to have so much variability due to environmental and 

### 4 colony dynamics. 

### 5 I mean, in theory it is possible, but I do not 

### 6 know. 

### 7 **DR. STEPHEN KLAINE:**Real quickly, just go to 

### 8 probably the easiest one or the one that you guys like, the 

### 9 tunnel study. Do you have enough information about variability 

### 10 in the tunnel study? You know, how many hives to put in? 

### 11 **DR. NANCY OSTIGUY:**I am going to disagree 

### 12 with Dr. Hunt a little bit. 

### 13 For particular outcome measurements I think we 

### 14 can do the power analysis just fine. It gets very difficult 

### 15 with some of the other measures or something that we really have 

### 16 not been talking a lot about. The things that we have suggested 

### 17 as possible future ways to analyze what is going on in honey bee 

### 18 colony, those we do not necessarily have enough information to 

### 19 do a proper power analysis. But colony survivorship, fecundity, 

### 20 those sorts of things. There is a tremendous amount of 

### 21 variability in the data but we can still do a power analysis. 

### 22 **DR. JEFF PETTIS:**There is a paper out there 

### 23 Rose et al., I do not remember the year; that was looking at 

### 24 effects of Bt corn pollen owned bees. And they did a power 

### 25 analysis, I think at the individual bee level, to look at sample 

### 26 size but also at the colony level, so there is a starting point.

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1 2 And then my other comment was going to be 

### 3 about the way to standardize colonies. We often will begin a 

### 4 group of colonies, just a group of 10,000 bees and a queen. We 

### 5 call it package bees. So they all start with the same starting 

### 6 population and presumably same disease levels and grow them for 

### 7 about a two month period. 

### 8 It is always advisable, if you need 40 

### 9 colonies for the study, start about 50 colonies; because as you 

### 10 began the study you can go through those and just take the ones 

### 11 that are of most equal strength. So there are ways to do that. 

### 12 13 **DR. GREG HUNT:**Unfortunately other sources of 

### 14 variability will be disease and honey bees have a lot of viruses 

### 15 with commonly asymptomatic viruses. If I went and tested my 150 

### 16 colonies now, they all have deformed wing virus. And the ones 

### 17 that have higher mite levels have bees that show deformed wings. 

### 18 If I get rid of the mites, they will still have deformed wing 

### 19 virus but they will look fine. 

### 20 One way to deal with that is to control your 

### 21 mites and to quantitatively measure your virus titers. I mean 

### 22 they could have three different viruses in them at one time so 

### 23 that is a source of variability. 

### 24 **MR. JENS PISTORIUS:**In addition, I think all 

### 25 has been said, but not by me. That's a saying we have. 

### 26 You made a very good point, Nancy, in saying

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1 that for most of the variables there are very good statistical 

### 2 tests possible with a high certainty. 

### 3 But on the other hand, for instance when you 

### 4 look at the number of colonies, when you are just looking at the 

### 5 certainty that you want to achieve, just look at the statistical 

### 6 numbers you would need, you end up with ridiculous numbers of 

### 7 colonies that are feasible to be tested. It is not feasible to 

### 8 test for every test, hundreds of colonies, basically just to get 

### 9 a statistical significance, but in my opinion this is not 

### 10 necessary at all to detect biological significance. 

### 11 So I think it has to be looked at with 

### 12 caution. Where you need really this statistical yes or no and I 

### 13 think we all agree that we cannot look at colonies just by 

### 14 evaluating statistics. 

### 15 So I think it is not appropriate to say, okay 

### 16 for power analysis reasons we need whatever, 40 colonies 

### 17 preferred because in other circles I know that such numbers have 

### 18 come up from statisticians who had no experience with bee 

### 19 trials. 

### 20 So I would like to warn that this may be a 

### 21 problem when communication the results. And I think that with a 

### 22 lot lower numbers preferred you already are able to do a good 

### 23 test. And probably this is also the same with semi-field test. 

### 24 You can do statistics, for instance, on mortality, but the most 

### 25 important thing is that you know a bit of bee biology, that you 

### 26 have expert judgement and you consider the biological

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1 significance. 

### 2 **DR. DAVID TARPY:**I just want to agree with 

### 3 all of those things but just to kind of step back to the 

### 4 question a little bit and say how we can focus and prioritize 

### 5 these uncertainties. 

### 6 I agree that we are never going to reduce them 

### 7 to real acceptable level without a ridiculously high sample 

### 8 size; you know all those kinds of things. And so I think what 

### 9 we need to emphasis from the answers from the previous question 

### 10 is that we are just going to have to rely on the interpretation 

### 11 of the given power of a given study and the inherent variability 

### 12 that is part of that study, that we can try to control by using 

### 13 sister queens and other means of standardizing, but we are not 

### 14 going to be able to get rid of all of them. We are not going to 

### 15 be able to get rid of all of the diseases. 

### 16 And so focusing on statistical power by 

### 17 increase sample size, replication of study, and trying to 

### 18 minimize the factors that we can, but looking at the important 

### 19 biological phenomenon at Tier IIs is really the issue. 

### 20 So I mean we can go on and on about all of 

### 21 these different things that need to be control, but in a 

### 22 practical way cannot always. So it is the accumulation of 

### 23 evidence that is important. 

### 24 **DR. NINA FEFFERMAN:**I normally do not suggest 

### 25 this especially for practical use because it is not a practical 

### 26 science, but everything that we have discussed in terms of

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1 significance seems to be boiling down to things like power 

### 2 analysis and p-values, which is a frequent, just statistical 

### 3 method. 

### 4 There are actually Bayesian methods for 

### 5 incorporating lack of understanding of variation in the system 

### 6 that give you, no longer a threshold test for "Yes this is 

### 7 meaningful; this is not", but an understanding of how meaningful 

### 8 given the amount of measurement and the amount of the inherent 

### 9 variation you expect for that measurement, how meaningful the 

### 10 interpretation of your measure have been. 

### 11 I do not want to go on record as suggesting 

### 12 that that become part of the standard expectation for these test 

### 13 but I would like to pipe up and say, "It is not the fact that 

### 14 statisticians are living in a black box and because they've 

### 15 never seen a bee, their work doesn't apply." 

### 16 There are methods for this - they are very 

### 17 good, the times that they have been used. They usually get 

### 18 used, honestly, at this point in practice only in things like 

### 19 human medicine or international politics that has nothing to do 

### 20 with science, kind of sociological studies to lend credence to 

### 21 some others position. 

### 22 But when these things have been applied, 

### 23 especially in the areas of risk assessment for then separate 

### 24 managerial decision making, they have proven to be very 

### 25 informative. So that may be something to keep in mind maybe in 

### 26 generations out - it would be a mistake in this conversation to

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1 assume that there do not exists techniques to handle this. 

### 2 **DR. DANIEL SCHLENK:**Just to follow up on that 

### 3 point. I do know having to attend SAP meetings recently that 

### 4 that is actually being proposed in risk assessment paradigm, 

### 5 Bayesian techniques actually. That is a huge discussion point. 

### 6 **DR. GREG HUNT:**Yes, Bayesian methods are used 

### 7 in many areas of biology. But getting back to the power 

### 8 analysis thing and agreeing with Jeff and Dave that with honey 

### 9 bee colonies, in Tier III studies, we cannot expect at the 

### 10 colony level statistical significance. But at Tier II and Tier 

### 11 III, if you can identify things that are important at the 

### 12 individual level, you can use statistics at the individual level 

### 13 to predict. 

### 14 **DR. ROSALIND JAMES:**Do you have a good 

### 15 general reference on how Bayesian might be used in a risk 

### 16 analysis, or analyzing this kind of data? 

### 17 **DR. DANIEL SCHLENK:**There will be a report 

### 18 coming out - well, the draft is being circulated right now, so I 

### 19 guess in another month, maybe? 

### 20 **DR. ROSALIND JAMES:**Can we put a reference to 

### 21 that in our notes? 

### 22 **DR. DANIEL SCHLENK:**You could, yeah. Yes. 

### 23 **DR. STEPHEN KLAINE:**There are a number of 

### 24 publications using Bayesian in terms of uncertainty analysis. 

### 25 Dwayne Moore and several others have published on this. 

### 26 **DR. ROSALIND JAMES:**That would be relevant

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1 for this type of experiment. 

### 2 **DR. STEPHEN KLAINE:**I'll get the references. 

### 3 4 **DR. MAY BERENBAUM:**It is a lit bit alarming 

### 5 to hear statements that you cannot do statistics on. I just 

### 6 want to make the observation that there appears to be a 

### 7 correlation, which is likely statistically between the amount of 

### 8 media interest in a particular subject and the number of poorly 

### 9 designed studies that lacks statistical analysis, which gets a 

### 10 disproportional amount of media attention. 

### 11 So I just want to emphasis that the absence of 

### 12 a statistical analysis or design that allows statistical 

### 13 analysis profoundly reduces the utility of any study, as it 

### 14 should. And this particular field is really prone to 

### 15 high-profile statistically - for want of a better word - 

### 16 worthless studies. 

### 17 **DR. JEFF PETTIS:**Just an additional comment 

### 18 about all of this. We convene Vets to talk about bee diseases 

### 19 and transmission of other animal diseases, and bees always come 

### 20 out as the exception. If you are dealing with dairy cow, 

### 21 chickens, or whatever, they are just so much easier to deal with 

### 22 because you do not have this individual to colony level effect, 

### 23 so this is nothing new, and in disease transmission around the 

### 24 globe and stuff. 

### 25 If you start dealing with any organism, a 

### 26 flying bird or a land animal, the things are all the same until

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1 you get to bees and then all the rules changes, so it is not 

### 2 unusual. 

### 3 **DR. DANIEL SCHLENK:**Okay, Dr. Tarpy a lot to 

### 4 integrate into that answer for you so I will leave that to your 

### 5 best judgment. And again let me just reiterate that if you are 

### 6 making comments, please get those written comments to Dr. Tarpy 

### 7 so that he can integrate that into the minutes. 

### 8 Any other panel comments? Okay, let me go to 

### 9 the agency, Mr. Sappington do you have any questions or 

### 10 clarifications. 

### 11 **MR. KEITH SAPPINGTON:**No Mr. Chairman we are 

### 12 good. 

### 13 **DR. DANIEL SCHLENK:**Great. Okay, let's move 

### 14 on to our last question, 14. You want to read that into the 

### 15 record. 

### 16 **MR. KEITH SAPPINGTON:**Charge Question 14a; 

### 17 please comment on the concept of using colony-level ecological 

### 18 models to inform the proposed risk assessment process for honey 

### 19 bees, as indicated above. 

### 20 **DR. DANIEL SCHLENK:**And our lead on this 

### 21 question is Dr. Fefferman. 

### 22 **DR. NINA FEFFERMAN:**So I'm kind of worry that 

### 23 everyone is going to hate me on this question. My basic answer 

### 24 to all three parts of this question is, each of these questions 

### 25 is important but I think you are starting with the wrong 

### 26 questions. So if you do not mind I am going to start with a

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1 sort of general explanation of what I mean by that and then move 

### 2 into answering the three particulars. Is that okay with the 

### 3 Chair? Okay. 

### 4 So to start with an idea of methodology and 

### 5 model choice that we have talked about a little bit earlier this 

### 6 week of analytic versus simulation models. And it seems that 

### 7 most of these questions have focus on the stimulation modeling 

### 8 level. 

### 9 And in my opinion this misses really where the 

### 10 greatest utility of modeling is going to be for these types of 

### 11 questions for risk assessment that are being asked throughout 

### 12 the rest of the White Paper. 

### 13 And that you guys have done an amazing job of 

### 14 picking out where the simulation type of questions have been 

### 15 addressed in the literature and where they might be able to help 

### 16 in the future but that that misses an entire analytic level. 

### 17 Especially for Tier I studies, I actually think that the need 

### 18 for simulation modeling might be obviated by the fact that - and 

### 19 this we will get to later in a sub-question -- by the time you 

### 20 have measured carefully enough all of the parameters that you 

### 21 would need and all of the outputs that you would then validate 

### 22 the model against, you may not need the model to answer the 

### 23 question anymore. And that for simulation modeling may for Tier 

### 24 I studies no longer then help you, although for Tier II and Tier 

### 25 III type equivalents would be very helpful. 

### 26 So to explain analytic models, they are kind

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1 of easy examples certainly from physics and I can detail some of 

### 2 those in the notes, although I probably will not go through them 

### 3 here. But the example that we discussed yesterday of the ratio 

### 4 instead of making the direct link from worker mortality for 

### 5 colony success to the ratio of worker mortality to worker 

### 6 eclosion rate was a very informative discussion from my end in 

### 7 terms of what an analytic model can be. 

### 8 That is an analytic model right there saying, 

### 9 "We need to look at that ratio." Now, of course that ratio is 

### 10 not all we need to discuss colony health; so for example notice 

### 11 that one worker dying over one worker eclosing for an entire 

### 12 colony would keep that ratio fairly healthy looking and that 

### 13 would be terrible. 

### 14 So we need to have minimum constraints on 

### 15 things like population sizes; and in some cases we need to have 

### 16 maximum constraint on population sizes to do things like if we 

### 17 want to prevent swarming or if we want to worry on a population 

### 18 level about carrying capacity type issues. 

### 19 But incorporating just those ideas of that 

### 20 ratio and that constraint without knowing any measurements 

### 21 already gives us sort of a universally applicable metric of 

### 22 colony health knowing if the colony is thriving or dying. And 

### 23 that is the hallmark of an analytically valid model, one that we 

### 24 can apply to gain insight regardless of the system. 

### 25 And actually Dr. James and I was just talking 

### 26 a little bit in the hallway afterward and I can put the details

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1 of that conversation in the notes, about how this could be 

### 2 tailored with the inclusion of one other variable to represent 

### 3 solitary bees as well and their success as a population instead 

### 4 of as an individual. 

### 5 And that means that we can get from this 

### 6 somewhat abstract. We are just talking about sort of workforce 

### 7 and its ability to provide for itself as a sustainable 

### 8 population. We can get very interesting insights and it is 

### 9 abstract enough to tell us where the detailed measurements need 

### 10 to go to inform that. 

### 11 So just off the top of my head right, if we 

### 12 say, okay there needs to be a minimum colony size -- that is 

### 13 something we would need to go measure. Is there a minimum size 

### 14 of the colony under which - and we do not actually need to 

### 15 measure it, we know that, but you guys see what I mean. 

### 16 So there are ways using these abstractions and 

### 17 equation-based things that give us thresholds. If that ratio of 

### 18 worker death to worker eclosion is less than one, the colony is 

### 19 going to die. There just is not replacement value for colonies. 

### 20 That is a strict threshold, that number is one. Right, if that 

### 21 ratio is less than one. That does not change regardless of what 

### 22 the worker eclosion rate is; so before we measure anything we 

### 23 now know something. 

### 24 So these are a way to get at some of the 

### 25 conversation we were having yesterday about how we identify the 

### 26 important things to go measure, where do we go look and the more

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1 generally applicability across these things. Of course then you 

### 2 sort of standardly validate them against expertise to ensure 

### 3 that you captured the relationships appropriately and they do 

### 4 still make simulation level predictions. 

### 5 From each analytic model you can then simulate 

### 6 an outcome. You probably can use an Excel program that captures 

### 7 that ratio and goes, "Okay, for a colony of this size should it 

### 8 do okay?" "How long should it take before it is not okay?" 

### 9 That is a testable hypothesis prediction that leads to more 

### 10 immediate approximate metrics for how to test, even if those 

### 11 metrics are not the goal endpoint. 

### 12 So it is actually one of the greatest 

### 13 utilities of this type of model in addition to providing those 

### 14 theoretical insights that you then generate incidental things to 

### 15 measure along the way that do not lead to the outcome of is the 

### 16 colony dying, but are still testable and measureable. 

### 17 And say, "Okay, if the colony is going to 

### 18 succeed there should be this other thing that has to happen as a 

### 19 logical necessity of that successful process. That is not an 

### 20 outcome we would discuss even normally a biologically relevant 

### 21 thing, but is a logical equivalent necessity of a good process 

### 22 happening. Where do we go measure those? 

### 23 This is not to say that this is the equivalent 

### 24 of a data gap. My saying that the questions and the White Paper 

### 25 do not address this type of modeling, that is not to say that 

### 26 there is a data gap here.

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1 One of the benefits of this analysis is that 

### 2 for non-risk assessment, non-pesticide and toxicity purposes, we 

### 3 have analytic models of healthy honey bee colony processes. 

### 4 I've written a couple of them and can provide references for 

### 5 that if that will be helpful, and I will just stick some in when 

### 6 I write up these notes. 

### 7 Each of those has mechanisms of impact from 

### 8 different pathways and it would be very easy in a modular sense 

### 9 to sort of unplug the pathway that is studied in that setting. 

### 10 Sometimes it is foraging richness and in fields there is 

### 11 uncertainty there. Sometimes it has been competition, sometimes 

### 12 it has been predators, any of these sets of things. 

### 13 We can modularly then plug in models that say, 

### 14 "Well, if the empirical measurement of toxicity says here is the 

### 15 impact on longevity, here is the impact on colony level 

### 16 fecundity, not necessary queen fecundity, but colony support of 

### 17 the eggs the queen lay. 

### 18 If we plug those in modularly to the numerator 

### 19 and the denominator, here is our outcome for is this colony 

### 20 thriving. And so there is not necessarily not at all forget 

### 21 necessarily. There is not a gap in the availability of these 

### 22 tools; they simply have not been applied directly yet to this 

### 23 question. So they have not been analyzed to say, "Well then if 

### 24 I just look at this model, the clear threshold would be blah." 

### 25 But they are easily calculable as soon as 

### 26 someone turns their mind to it. And as soon as someone has some

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1 of the individual and group level measurement that you guys are 

### 2 discussing getting already, without that necessarily being the 

### 3 end point. Where we could use those as proximate ways to get at 

### 4 the ultimate question of what is the success of a colony and 

### 5 what is the success of a population at providing pollinator 

### 6 services, at being sustainable itself and having population 

### 7 level success. 

### 8 Lastly, in the general sense - sorry I realize 

### 9 this is a very long answer -- in the general sense modeling does 

### 10 create one set of complications that nothing else we have 

### 11 discussed creates for the levels of the Tiers. It is important 

### 12 to point out that the tiers themselves are somewhat problematic 

### 13 in the sense that the kind of analytic modeling that I am 

### 14 suggesting would be useful. 

### 15 Because colony level models should ideally be 

### 16 used backwards to inform which Tier I types of studies should 

### 17 happen to be most useful as a pass; by which if you pass Tier I 

### 18 and you do not set off any alarms to trigger further study, 

### 19 those were your important metrics. Those need -- not semi-field 

### 20 and field, in that sense I do not mean tier, but because of the 

### 21 way that these study works we have been discussing Tier II and 

### 22 III models as being the ones that leads to colony level and 

### 23 population level type questions, because those are hard to get 

### 24 to in a lab. 

### 25 So that equivalence of which tiers mean which 

### 26 level of scale, no longer applies within the modeling. And a

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1 Tier I study should possibly be, even a population of colonies 

### 2 level model in order to backwards inform what the individual 

### 3 honey bee or solitary bee metrics should be that are most 

### 4 critical to indicting health, and I mean health in a 

### 5 sustainability kind of sense. 

### 6 Very briefly then to address the specific 

### 7 question of (a) which was the concept of colony-level ecological 

### 8 models, yes it is great. The short answer is we should do it. 

### 9 We should do it in intelligent ways. So exactly the types of 

### 10 questions that we have been saying are very hard to get to 

### 11 experimentally of how do individuals scale to colony, scale to 

### 12 populations, and how does all of that go as things fluctuates 

### 13 over time both in terms of natural stochastic fluctuations and 

### 14 in terms of bees changes. 

### 15 The year changes, bees changes. The need 

### 16 changes; bees have different behaviors if a bear comes by, that 

### 17 kind of stuff; although agricultural bears are probably not a 

### 18 big problem, but still. 

### 19 So these kinds of models are simulation and 

### 20 analytic but especially simulation, the kinds of models you 

### 21 proposed directly in the paper, are going to be quite useful for 

### 22 doing that. All this being true, there is a huge pitfall to be 

### 23 avoided in doing this. It would be a mistake to just suppose 

### 24 that the best modeling practice for colony-level ecological 

### 25 models to inform risk assessment would be an all-inclusive, it 

### 26 has every aspect of colony function and every risk model. That

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1 is a bad way to model this even in simulation. 

### 2 And that has to do both with being able to 

### 3 interpret the outcomes and knowing which factors were driving 

### 4 them but also as we just addressed in some of question 13, there 

### 5 is a confounding uncertainty from each modular plug-in. And 

### 6 that is both the uncertainty of the measurement and the 

### 7 uncertainty of the stochasticity. 

### 8 And there are really intricate ways in which 

### 9 those interact and there are some tools from statistical 

### 10 mechanics to try and correct for that, that we do not want to 

### 11 use for this. 

### 12 And so a much better way of approaching an 

### 13 all-inclusive type of model is to have littler models as some of 

### 14 are in the White Paper discussed, and then have a risk assessor 

### 15 as a person with expertise decide how the outcomes from those 

### 16 really might interact. Sticking them all into one model where 

### 17 the math comes out as one number is a great way to confuse 

### 18 ourselves. 

### 19 That is basically my answer to a. 

### 20 **DR. ROSALIND JAMES:**So my role on this team 

### 21 was to help Nina give examples where things were not clear. So 

### 22 I am going to ask you a couple of questions that I had. 

### 23 When you are talking about the tier test you 

### 24 said it slightly backward. I wanted to make sure you understand 

### 25 in the tier testing things move to higher tiers when they fail 

### 26 the lower tier, right?

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1 **DR. NINA FEFFERMAN:**Sorry, I misspoke. What 

### 2 I mean is if there is a trigger that says I failed Tier I. 

### 3 **DR. ROSALIND JAMES:**So then a higher risk, 

### 4 right, not less risk? 

### 5 **DR. NINA FEFFERMAN:**Yes, right. 

### 6 **DR. ROSALIND JAMES:**Okay, I wanted to make 

### 7 sure that you understood that. 

### 8 And so I was not exactly sure what you were 

### 9 saying regarding the tier-level and maybe to help clarify I will 

### 10 put it in another question. Could you go back and look at 

### 11 current colony-level data that we already have for pesticides 

### 12 that are in the literature to go back and help determine what 

### 13 Tier I factors we should note? 

### 14 **DR. NINA FEFFERMAN:**Thank you Dr. James so 

### 15 much. Yes, that is exactly what I meant we should be doing. 

### 16 That we should take the colony-level understanding that we have 

### 17 and any future colony-level understanding from whichever either 

### 18 Tier II or Tier III studies are proposed, if things triggers 

### 19 higher concerns from Tier I; to go back to suggest further 

### 20 refinements in which Tier I should trigger further concern. 

### 21 Does that help clarify? 

### 22 **DR. ROSALIND JAMES:**I think so, but could you 

### 23 use an example? 

### 24 **DR. NINA FEFFERMAN:**For example, and this is 

### 25 a silly example because it will wind up seeming obvious but 

### 26 there are others. Actually, can I depart from honey bees

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1 comfortably for an example? 

### 2 Okay, awesome. So we have been talking about 

### 3 some desert tortoise conservation issues and doing some of the 

### 4 desert tortoise conservation work - I was helping model some of 

### 5 this -- we went initially to a bunch of desert tortoise experts 

### 6 and said what do you think are the most salient features of the 

### 7 endangered population of desert tortoises we are working with. 

### 8 And they gave us a list of here are the ones we think are really 

### 9 important and then here are the ones that we think are sort of 

### 10 ancillary but tortoises depend upon these things. 

### 11 By looking at the population level 

### 12 determinants of these factors we were able actually to move some 

### 13 things out of the categories that the experts expected were 

### 14 going to be very important. Saying, okay, yes that is clearly 

### 15 killing tortoises; that is not the thing driving population 

### 16 decline. That is some of the things you are seeing when you go 

### 17 find a dead tortoise; that is what you are seeing the dead 

### 18 tortoise do. But it turns out that the value of that age-class 

### 19 to the demographic process of population growth, it is not 

### 20 valid. 

### 21 On the other hand, there were some things that 

### 22 were unexpectedly incredibly important. Water reservoir 

### 23 balance, if you have a respirator infection as a tortoise it is 

### 24 apparently really important for conservation. And water balance 

### 25 was known to be important. Respiratory infections were not 

### 26 necessarily. And the fact that you get a runny nose turns out

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1 to drastically change your water balance. 

### 2 Fugitive dust was assumed to be a very 

### 3 important thing. Fugitive dust, you have an ATV kick up dust in 

### 4 the desert and then it doesn't settle for a while, turns out 

### 5 basically unimportant for our population. 

### 6 So is that a kind of concrete example where, 

### 7 from going from the population level in modeling, what is the 

### 8 demographic growth process of that population. 

### 9 We were then able to go back to these 

### 10 individual measurable things either about the environment or 

### 11 about the individual tortoise. And say okay, these are the 

### 12 things you should measure. If you want to be concerned about 

### 13 your population or if you want to be confident your population 

### 14 is okay. 

### 15 Does that clarify? 

### 16 **DR. ROSALIND JAMES:**So here is the case where 

### 17 you could go back. Now the next endangered species. What did 

### 18 you learn form that that you can say now for Tier I test we want 

### 19 to measure? 

### 20 **DR. NINA FEFFERMAN:**So the process of that 

### 21 modeling is the thing that ports across species. But it would 

### 22 be true that we would reformulate the models themselves if we 

### 23 went from - I mean, if we look at something that is still like a 

### 24 tortoise then it would still apply. Sure, for other tortoises 

### 25 not the desert tortoise. 

### 26 Then we could re-apply the same logic and use

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1 the same parameters and that does give us thresholds for which 

### 2 measurements. So we could say, well when we stick in the value 

### 3 for -- here is for example the non-stable age demography. It 

### 4 turns out that many of the models -- the outcome is very 

### 5 sensitive to how many juvenile you have versus how many 

### 6 reproductive adults. And it turns out that the size of an adult 

### 7 female is a really important thing for tortoises because eggs 

### 8 have to fit under the shell. 

### 9 But that turns out to be important in the 

### 10 demography we had for our population. We would reapply that 

### 11 analysis in a new population and say well how many adult 

### 12 tortoises do you have? If you have zillions of adult female 

### 13 tortoises, then female tortoises' mortality might not be the 

### 14 issue; it might be egg survival. And that is putting in new 

### 15 number to the same analysis to get out a different insight for 

### 16 that population, or for that species. 

### 17 That model would not directly apply to 

### 18 colonies of honey bees but the exact same methods would and we 

### 19 do have colony and population-level models of honey bees that do 

### 20 equivalent things. It is just that since I have not done them 

### 21 in this context of toxicology I do not have as ready yet, an 

### 22 answer for like, "this is a surprising result I expect to get 

### 23 out" because if I expected it, it would not be surprising. 

### 24 **DR. DANIEL SCHLENK:**Okay, let's go to Dr. 

### 25 Potter. 

### 26 **DR. THOMAS POTTER:**Well I certainly agree

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1 with my colleagues and concur that it is important to have a 

### 2 multi-dimensional approach to the problem, if I am correct in 

### 3 paraphrasing here. And certainly there are many different types 

### 4 of models that can be used, again both analytic and simulation. 

### 5 6 To further try to compartmentalize this 

### 7 problem, which obviously is exceedingly complex, we can -- again 

### 8 as Nina mention there are modular approaches that can be used 

### 9 and at minimum I believe we need to step back and say you know 

### 10 this is a two-part problem. 

### 11 And forgive me for stating the obvious, but it 

### 12 is exposure and response. So I would say that as a first cut we 

### 13 divide our problem down into two parts, number one exposure and 

### 14 response. 

### 15 Exposure is probably more amendable to 

### 16 simulation modeling I would hazard a guess, maybe, maybe not. 

### 17 It certainly has a long history in all of the other aspects that 

### 18 the Office of Pesticide Programs addresses in terms of 

### 19 evaluating exposure; certainly simulation models are critical 

### 20 tool, certainly on the exposure side. 

### 21 And so I strongly would emphasize that effort 

### 22 be directed in that area to develop simulation models, whether 

### 23 they be pesticide Environmental Fate model that works for a 

### 24 colony in terms of residues that are in the colony and 

### 25 determining what their fate may be in terms of persistence, 

### 26 longevity, whatnot. Obviously those are critical questions that

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1 need to be answered in terms of exposure at different 

### 2 life-stages, et cetera, overwintering success. 

### 3 And then the other part of that problem, and I 

### 4 think again Dr. Fefferman identified this and I think it is 

### 5 critically important and an exceptionally useful tool, perhaps 

### 6 at Tier I, and that would be the use of inverse modeling to try 

### 7 to do some ground-truthing, particularly in the side of the 

### 8 exposure pathway. 

### 9 Again, obviously as has been discussed over 

### 10 the last few days, we have some uncertainty about exposures. 

### 11 Efforts have been made to identify where those are. One 

### 12 possible approach to get at this problem is to take existing 

### 13 colony data and do adverse modeling; looking at the different 

### 14 compartments where residues are stored within the colony and 

### 15 inverse model to see if there are insights to be gained on where 

### 16 those residues may have come from, whether they are potentially 

### 17 dust emission. 

### 18 Again, there are some timing issues and 

### 19 whatnot here et cetera, whether they came in by water or they 

### 20 are totally dietary. I think to some degree we can gain insight 

### 21 by going through that process. 

### 22 I will just sum up that that is obviously a 

### 23 big problem. We recognize that and we would benefit 

### 24 substantially by subdividing it and proceeding forward from 

### 25 there. 

### 26 **DR. DAVID TARPY:**I agree with and very much

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1 appreciate this discussion and really have nothing more to add 

### 2 other than to emphasize the previous comments that were made 

### 3 that colony-level modeling is very helpful in a kind of a 

### 4 top-down approach. I think the statement was made earlier that 

### 5 predictive modeling from lower tiers to upper tiers is something 

### 6 that we need to be very cautious about. 

### 7 And so just simply taking information from 

### 8 sub-colony phenomena and trying to model that upwards to colony 

### 9 and population-level is froth with potential problems; rather 

### 10 taking a colony-level top-down approach of trying to understand 

### 11 that phenomena that informs the lower tiers is going to be much 

### 12 more effective. 

### 13 **DR. DANIEL SCHLENK:**Other panel members. 

### 14 **MR. JENS PISTORIUS:**1:02:08 I wish I had only 

### 15 a fraction of understanding of modeling of what you guy do. I'm 

### 16 really impressed and my brain is working a lot at the moment and 

### 17 I might be confused so excuse me in advance if this answer may 

### 18 sound funny and some things may have been said. My answer will 

### 19 be short. 

### 20 Yes, to inform and to identify potential risk, 

### 21 which trigger maybe the need for higher tier tests; but I want 

### 22 to point out that, no for if you have a model that tells you if 

### 23 you identify a risk in Tier I, and then you come up with a model 

### 24 calculation that said well you do not need a Tier II and Tier 

### 25 III, I think this would be a funny way you would have two 

### 26 complimentary ways in risk assessment, which somehow does not

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1 fit in a risk assessment scheme. 

### 2 **DR. NINA FEFFERMAN:**Mr. Pistorius, I am 

### 3 probably misunderstanding. I think that what you just said is 

### 4 that you should only use modeling for what to do in Tier I if it 

### 5 agrees with the empirical work. That cannot be what you meant. 

### 6 **MR. JENS PISTORIUS:**I'm not quite sure if I 

### 7 understood because again my brain might be overworked at the 

### 8 moment. 

### 9 I just want to say, if you identify a risk in 

### 10 the known Tier I studies and if you have this risk assessment 

### 11 process as it is outlined now and if you trigger higher tier 

### 12 tests. And then if you come up in parallel with the modeling 

### 13 that says, okay but all you have done in your Tier I 

### 14 identification of a potential risk is all not valid. And then 

### 15 you basically have to decide okay our Tier I process identified 

### 16 would have to go to a higher tier study. 

### 17 And then on the other hand you come up with a 

### 18 model that says, well yes you have identify the risk in Tier I 

### 19 but actually it is not necessary, you do not need to do any 

### 20 further studies. That is something I would reject because then 

### 21 you would have two parallel ways in risk assessment. 

### 22 Was that clear and understandable? 

### 23 **DR. NINA FEFFERMAN:**I think I understand you 

### 24 but disagree with you. I think that in my mind that is exactly 

### 25 the use of half of one of the ways that this works. 

### 26 One would be to identify Tier I things that

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1 should be measured empirically that would then trigger the need 

### 2 for higher studies by concern; but the other would be to show 

### 3 when we think things that need to be measured that would trigger 

### 4 higher concern should actually not effect success. 

### 5 And if we reject that half of the modeling 

### 6 then I do think that we disagree. I think that is still a use 

### 7 for modeling. 

### 8 **DR. MAY BERENBAUM:**Good luck explaining that 

### 9 to the public. That is an issue that exactly addresses your 

### 10 point that if there is a cause for concern generated by 

### 11 empirical data at Tier I and a decision is made based on models 

### 12 not to proceed, at lease for the American public, that would I 

### 13 think create a major concern. 

### 14 **MR. JENS PISTORIUS:**I think it is the same 

### 15 for the European public but also for me as a person 

### 16 understanding. Sorry that I really have to say I disagree. 

### 17 I want to see studies with bee colonies and 

### 18 because we have identified that bee colonies are a lot more 

### 19 complex than the individual and modeling may be very good and 

### 20 may inform a little bit, but if this decision is to be made and 

### 21 say, okay you have identified a risk with a normal risk 

### 22 assessment Tier I process, if model comes up to say, well you do 

### 23 not have to do higher tier studies, I would clearly reject that. 

### 24 **DR. NANCY OSTIGUY:**I think that this 

### 25 discussion points out the importance of doing the analytical 

### 26 models first. And we really should do those first so we can

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1 then target which Tier I studies are the most important to do, 

### 2 so we do not have contradictory conclusions that we have to 

### 3 resolve. 

### 4 **DR. MARTHA SANDY:**Just to remind everyone, 

### 5 risk assessment is -- if you remember, the framework diagram we 

### 6 saw - it is supposed to be an iterative process. And I think 

### 7 that that is where we have some confusion going on. 

### 8 I think what Nina is suggesting is, we have 

### 9 come up with some Tier I endpoints we think are important to the 

### 10 colony. She is suggesting we use the analytical models to see 

### 11 if those endpoints in fact are relevant to colony health. And 

### 12 she gave the example using a different species. 

### 13 But to take another example from the honey 

### 14 bee, what if we think mortality to drones is an important thing 

### 15 but in the model it turns out that that does not seem to be a 

### 16 driver to colony health but what does seem to be a driver is 

### 17 mortality to larvae and length of time of development of larvae 

### 18 for example; those are really key. 

### 19 Then what you would do is you would take that 

### 20 information from the model, once you felt confident that you 

### 21 could rely on it, you might go back and revise the Tier I test 

### 22 requirements with which things in Tier I endpoints the 

### 23 measurements that you place and emphasis on that triggered going 

### 24 to the Tier II or Tier III test. 

### 25 So I think you are talking at cross-purposes 

### 26 because you are not, right now Mr. Pistorius, ready to accept

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1 that the modeling has been validated and everyone has agreed to 

### 2 change the current Tier I endpoints that are measured. 

### 3 We are not there yet; we have not even done 

### 4 the modeling. But, there would be a period of time of 

### 5 discussion and then different groups would decide whether they 

### 6 still want to evaluate a certain measurement. 

### 7 **MR. JENS PISTORIUS:**Well I think such 

### 8 modeling are beautiful for the weight-of-evidence and the 

### 9 multiple information that you need for the conclusion on the 

### 10 risk assessment; on the other hand from my experience it is not 

### 11 that an applicant comes with one study and then you get concern. 

### 12 On the Tier I level, usually what we get is a package of the 

### 13 standard requirement, which are in our case oral contact and 

### 14 maybe in future the larvae if we are able -- or larvae will be 

### 15 addressed as we get there. 

### 16 So that is basically the standard package we 

### 17 ask for to be able to make a first conclusion. And then, okay I 

### 18 admit maybe such models may inform do you need other studies on 

### 19 the tier level one risk assessment. But still on the first 

### 20 three that you get, on the contact, oral and larvae, you already 

### 21 do make a decision, you know, are we having a concern? 

### 22 And so from my understanding, but I may be 

### 23 completely wrong, that is already triggering something. And I 

### 24 am really glad if you say as a risk assessor, okay we have 

### 25 identify risk in Tier I, we go to Tier II and we ask for 

### 26 semi-tier study and we find no effect.

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1 And then you say okay for our 

### 2 weight-of-evidence you take those multiple information into 

### 3 account and then you can say with addressing the uncertainties 

### 4 that you get in your risk assessment and say okay with this 

### 5 weight-of-evidence we have the Tier I, Tier II and then we find 

### 6 no effect and this is basically also what the colony said, so I 

### 7 think you have a lot of weight. 

### 8 That is my simple understanding of the issue. 

### 9 **DR. NINA FEFFERMAN:**Thank you very much Dr. 

### 10 Sandy. I think the iterative process is what I have in mind and 

### 11 it sounds to me like what you are suggesting is that you would 

### 12 trust modeling to suggest additional test but not to eliminate 

### 13 existing ones. And I think then we just have a difference of 

### 14 opinion on the utility of models. 

### 15 But to address Dr. Berenbaum's comment about 

### 16 could the public ever accept this, the answer in my mind is 

### 17 salesmanship. We have a couple of examples where we botched it 

### 18 entirely and a couple of examples where we have done it really 

### 19 well. 

### 20 We botched it entirely with vaccine safety; we 

### 21 have also botched that with empirical studies. So just having 

### 22 empirical studies is also not a guarantee of public acceptance 

### 23 of the results. 

### 24 But we have done it really really well with 

### 25 some of the U.S. Fish and Wildlife Services Conservation 

### 26 Endangered Species Act requirements. And when you should close

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1 roads that inconvenience people in order to left spotted 

### 2 salamanders cross and things like that. And we have had really 

### 3 good public acceptance and that spotted salamander work, all 

### 4 modeling, all of it. 

### 5 And it originally met with huge public 

### 6 resistance and we did really good outreach. And we went in and 

### 7 we said, okay guys this is what went into this. We did not 

### 8 explain the equations but we explained the conceptual flow of 

### 9 what we were modeling and we now have communities that come back 

### 10 to us every year and go, "Should we be closing the same roads at 

### 11 the same time this year?" 

### 12 And so my answer to that part is, that is a 

### 13 matter of public outreach and effective communication. It is a 

### 14 slightly harder sell to sell the public on modeling versus 

### 15 empirical work, but forgive me, I have great faith in the 

### 16 public; I do not have great faith in the average person as a 

### 17 scientist. 

### 18 So I think we have work to do to explain 

### 19 empirical science to the public, also we just have a little bit 

### 20 more work to do to explain mathematical science. 

### 21 **DR. THOMAS POTTER:**I just want to make a 

### 22 comment. I'm not sure I heard anywhere in the discussion, 

### 23 certainly in Dr. Fefferman's answer at the outset as well as the 

### 24 other discussants, that we as a group or anyone was recommending 

### 25 that modeling be used to in some way subvert or eliminate or in 

### 26 otherwise change the tier risk assessment process.

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1 We have Tier I, if you fail Tier I one assumes 

### 2 then you get to Tier II. And certainly, this again, there is a 

### 3 long history of that in terms of looking at other problems that 

### 4 the agency deals with. I appreciate your concerns that somehow 

### 5 an ill-informed model may somehow obscure the need for further 

### 6 testing but I do not think that that was in any way suggested. 

### 7 **DR. JAMES MCMANAMAN:**So I got really 

### 8 confused. I really appreciate your discussion Nina. 

### 9 It appears to me that (a) is asking a 

### 10 different kind of question than what we have really been 

### 11 discussing. Because if we take your examples of analytical 

### 12 modeling - and I may be way off base here -- if you take your 

### 13 examples of analytical modeling and apply them to what are the 

### 14 causes of loss of bees, I think that the European data would 

### 15 suggest that pesticides are a cause of loss of bees. 

### 16 If the question is then how should we model 

### 17 pesticide effects on bee colonies, then it seems to me it is a 

### 18 different kind of modeling process all together. Is that not 

### 19 the case? Because in one case you are asking, is it a risk 

### 20 factor, and if the answer is no, then we have eliminated the 

### 21 entire question. So if the answer is no, but we still want to 

### 22 know more about how pesticides affects bees, then I think it 

### 23 seems to be more of a different type of modeling rather than 

### 24 analytical type. 

### 25 **DR. NINA FEFFERMAN:**Thank you. That is a 

### 26 wonderful clarification question.

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1 When I was talking before about colonies 

### 2 thriving or dying as an outcome, I should have been more 

### 3 explicit. I meant that restricted to the example I was using 

### 4 from the discussion from yesterday, of what is the importance of 

### 5 a particular threshold level in worker mortality. 

### 6 But I do mean to suggest that analytic models 

### 7 themselves are going to directly and validly, in risk 

### 8 assessment, be able to point to us the most important either 

### 9 that variables or functional relationships or thresholds for 

### 10 where pesticides can affect success by whichever metric we 

### 11 adopt, either pollinator services or survival of the colony or 

### 12 sustainability of the population. 

### 13 At any level of those analytic models will be 

### 14 able to suggest to us -- for it to be a good model both testable 

### 15 predictable outcomes that should be before we get to the outcome 

### 16 that we are interested in so that we can validate the model and 

### 17 not necessarily on an agreed outcome of what we want but 

### 18 proximate measures that should be easy to measure also easy to 

### 19 see when the answer is negative. They should not be patently 

### 20 obviously always going to be true. 

### 21 So a way of validating, if you would, the 

### 22 by-products of the model as a way to have confidence in the 

### 23 entirety of the model when we are asking these difficult 

### 24 questions; but then apply it to gain insight into which things 

### 25 should be measured that will directly affect whether or not a 

### 26 pesticide is posing a risk to our outcome. Thank you so much.

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1 **DR. JAMES MCMANAMAN:**Thanks for the 

### 2 clarification. 

### 3 **DR. DANIEL SCHLENK:**Okay, any other comments 

### 4 right now? I'm going to try to bring us back hopefully. It 

### 5 seems like all three of the questions have sort have been 

### 6 discussed at the same time interactively here. So let's go back 

### 7 to (a) and just say, please comment on the concept of using 

### 8 colony-level ecological models to perform proposed risk 

### 9 assessment for honey bees. 

### 10 Okay so we have had comments, and it appears 

### 11 that in general the panel feels, yes definitely use models. I 

### 12 think there is some disagreement perhaps in what outcomes those 

### 13 models could be used for, perhaps, so let's make sure we get 

### 14 that into the minutes in terms of concerns for certain panel 

### 15 members and others. 

### 16 But it would appear that everybody is 

### 17 comfortable with the concept of using models for this particular 

### 18 parameter. Is that what I'm hearing? Yes, okay. 

### 19 All right, any other comments before we ask, I 

### 20 am sure there is going to be questions of clarification from the 

### 21 agency, I am guessing. Any other comments before we go to the 

### 22 agency? 

### 23 **DR. THOMAS STEEGER:**I would just like to make 

### 24 a comment and I appreciate the dialog that is going on regarding 

### 25 how you interpret information across multiple levels of 

### 26 biological organization.

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1 The National Research Council through the 

### 2 National Academy of Science in their publication, Toxicity 

### 3 Testing in the 21st Century, has sent a strong message that 

### 4 there should be movement away from whole animal testing and 

### 5 taking advantage of as much of the available information as you 

### 6 can so that you can move from one level of biological 

### 7 organization to another without as much reliance as we have in 

### 8 the past on whole animal testing. 

### 9 And that particularly deals with the screening 

### 10 level approaches where that would enable of regulatory authority 

### 11 to determine where you need to best use resources and apply 

### 12 them. 

### 13 And this whole idea that we touch on in the 

### 14 paper, adverse outcome pathways, expert systems, that once those 

### 15 relationships become clear and you can, based on the mode of 

### 16 action of a chemical, you can have a mechanistic approach that 

### 17 would enable you to make some decision that would be less 

### 18 reliant on animal testing, is the direction that this agency I 

### 19 believe is headed. 

### 20 But with that said, where you have identified 

### 21 risks you would be transitioning likely to higher tiered test to 

### 22 substantiate these potential concerns. Because companies do put 

### 23 a lot of money into the development of chemicals and it is 

### 24 ultimately up to the risk manager to decide where a higher tier 

### 25 testing is required but it is also the purview of the regulated 

### 26 community to demonstrate their product is safe as it is actually

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1 planned to be used. So the higher tier testing would still have 

### 2 a role. 

### 3 But I applaud the efforts to introduce this 

### 4 idea of using information at multiple levels of biological 

### 5 organization to inform he risk management decision. 

### 6 **DR. DANIEL SCHLENK:**Okay, so with that I am 

### 7 assuming you got what you need for this particular letter. Is 

### 8 that correct? Okay. With that let's just take a break I think 

### 9 before we go on to (b) and (c). 

### 10 (Brief recess.) 

### 11 **DR. DANIEL SCHLENK:**Okay, just to remind the 

### 12 panel, during breaks if you do have any interactions with the 

### 13 audience or the agency just to make sure that we state that for 

### 14 the record that you have had those interactions and if it has 

### 15 changed your comments, you need to state that for the record as 

### 16 well. Just to let you guy know we could do that. 

### 17 **DR. MAY BERENBAUM:**Do you want all 

### 18 interactions recorded or only those that have any bearing on the 

### 19 discussion. 

### 20 **DR. DANIEL SCHLENK:**Those that affect your 

### 21 comments please. 

### 22 **DR. ROSALIND JAMES:**Do you want those now? 

### 23 **DR. DANIEL SCHLENK:**I will give you the 

### 24 opportunity as soon as we get to the questions, yes. Let's go 

### 25 ahead and read letter (b) into the record. 

### 26 **MR. KEITH SAPPINGTON:**Dr. Schlenk, if I could

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1 ask one clarifying question. 

### 2 I think it would be helpful to the agencies in 

### 3 responding - and I'm not sure which of these questions or all of 

### 4 them - about the difference between an analytical model and a 

### 5 simulation model and perhaps a concrete example, and perhaps 

### 6 when one may be preferred over another. 

### 7 **DR. DANIEL SCHLENK:**Dr. Fefferman. 

### 8 **DR. NINA FEFFERMAN:**Thank you for asking that 

### 9 question and actually we did have a useful conversation on 

### 10 exactly that point during the break. So I will bring up all of 

### 11 those points but this is my active disclosure on that one. Do 

### 12 you want me to answer or just put it in the minutes? 

### 13 **DR. DANIEL SCHLENK:**No, I think just the fact 

### 14 that you can do that would be appropriate. 

### 15 **DR. NINA FEFFERMAN:**Oh, okay. Yes, we can do 

### 16 that. 

### 17 **DR. DANIEL SCHLENK:**Okay, ready to go on to 

### 18 letter (b)? 

### 19 **MR. KEITH SAPPINGTON:**Question 14, letter 

### 20 (b); please comment on the state of the science regarding 

### 21 available honey bee models discussed in Section 5.4.2 of the 

### 22 White Paper in relation to the potential application in a 

### 23 regulatory risk assessment context. In particular, please 

### 24 comment on the extent that such models have been evaluated using 

### 25 empirical data related to honey bee population dynamics and the 

### 26 availability of such data for their parameterization.

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1 **DR. NINA FEFFERMAN:**I going to rely on the 

### 2 answer to the last question, which is going to come down to for 

### 3 analytic models data is less important than for simulation 

### 4 models. And with that context in mind many of the models 

### 5 mentioned in the White Paper have only very sparse assess to 

### 6 data thus far. And we have discussed during this panel many 

### 7 good ways to try and fill some of those data gaps. 

### 8 And while it is very important to collect that 

### 9 further input data, for measurements, for parameter values, and 

### 10 for model validation and to characterize functional 

### 11 relationships. Many of these analytic models are useful in the 

### 12 absence of direct measure data as long as the functional 

### 13 relationships themselves are hypothesize correctly by the 

### 14 experts building them. 

### 15 And so this does not happens in a vacuum of 

### 16 data but what happens is that there is empirical work that 

### 17 informs relational data. So how do we get - on a very stupid 

### 18 level - how do we get a bigger colony? We have more babies than 

### 19 we have deaths, right? That is something that at some point 

### 20 someone measured and went, "Oh look, demography, demography 

### 21 happens" when there are more births than death. From that you 

### 22 get an abstract analytic model, okay births has to outweigh 

### 23 deaths for a population to grow; the equivalent for that has to 

### 24 happen for a colony for worker mortality to worker eclosion. 

### 25 That is a valuable insight; that one is a 

### 26 trivial insight but valuable insights come out of that

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1 relationship in the absence of, what is the worker mortality 

### 2 today; or what is the overall colony survival with that. 

### 3 So the conclusion of these models to provide 

### 4 insights into how the systems work and to make specific 

### 5 predictions about which focal metrics are important, are really 

### 6 the utility of the analytic side for purposes of risk 

### 7 assessment. The more general employment of models that can help 

### 8 assess the importance of unknown levels of threat involves both 

### 9 the analytic and the simulation levels of these things. 

### 10 So even if very little insight is known about 

### 11 individual sensitivity to -- for instance exposure to particular 

### 12 chemicals what you can do is know which pathways to effect, 

### 13 either within the analytic models that are more abstract or the 

### 14 empirical models that are the specific. Okay, the link is 

### 15 multiply this number by .003, get that number that is the 

### 16 answer. 

### 17 Within the narrow context of the empirical 

### 18 models discussed in the White Paper, the simulation type models 

### 19 -- in modeling, simulation models are also called empirical 

### 20 models, which is not to be confused with empirical experiments. 

### 21 Sorry, I am going to try and remember not to call them that. 

### 22 Simulation experiments, in the narrow context 

### 23 of the simulation models being used we then need to discuss the 

### 24 appropriate sensitivity of the data that is being used, the 

### 25 sensitivity and robustness testing for the individual 

### 26 parameters. So there is sensitivity which is how do the value

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1 affects the ultimate outcome and there is robustness, which is 

### 2 what are the range of values for which this model is still 

### 3 valid. 

### 4 And for a simulation model we have not 

### 5 discussed robustness at all. There are very good local range 

### 6 simulation models that tell you exactly as long as PH does not 

### 7 go off the scale everything in the system behaves exactly this 

### 8 way; but as soon as you have a PH under three, oh my God do not 

### 9 use this model; use a different model. 

### 10 There is a different behavior, in physics it 

### 11 is called a phase transition, right, something else has 

### 12 happened. You have coalescence or rain from a cloud, right. 

### 13 There is water, suddenly something happens and water falls out 

### 14 of the cloud. A model for how water is suspended in the air 

### 15 works really well up until a point, and then you need a model 

### 16 that says okay how does rain falls. And I am not a 

### 17 meteorologist so they might actually have a good model that does 

### 18 both of those; I do not know. 

### 19 So within the available data that we have 

### 20 first we have to ask sort of what do we want out of the model; 

### 21 because if the answer is most appropriately answered by an 

### 22 analytic model, we may not need any of the data; what we want is 

### 23 the insight and that will tell us which data then is going to be 

### 24 important, and then we know whether or not we have it. But we 

### 25 do not know whether or not we have it until we have that 

### 26 analytic model to tell us.

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1 For a simulation model we do have to go 

### 2 through the sensitivity of each of the values that we would need 

### 3 and say, well what if this was everywhere from here to here. 

### 4 Does that change the outcome at all? If the answer that comes 

### 5 out of the entire model of the simulation does not change with 

### 6 regard to the quality of the data input for that parameter, you 

### 7 do not need a better measurement of that parameter. On the 

### 8 other hand if it is very sensitive to that parameter, you need a 

### 9 very good set of empirical experiments to do that. 

### 10 So unfortunately there is not an easy answer 

### 11 to, oh for all of the ones in the White Paper here is the data 

### 12 gap, here is what we know; we are good just use those. Go 

### 13 measure those and at the end we will be fine. On the other hand 

### 14 there is sort of an algorithmic approach that we can take with 

### 15 each parameter for each model going, is it sufficient; is our 

### 16 knowledge good enough to use that with confidence in this 

### 17 context. 

### 18 **DR. ROSALIND JAMES:**I'm debating whether just 

### 19 to say I concur or try to elaborate more. I'll try to give a 

### 20 little example maybe. 

### 21 In the White Paper I think the review of 

### 22 simulation model is excellent and fairly complete; there may be 

### 23 some missing models but I think it is a fairly complete review 

### 24 and a nice summary of the model. 

### 25 An example I think of what Nina is talking 

### 26 about is say with the Khoury et al model, we discussed this

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1 earlier and one of the conclusions was there is this critical 

### 2 threshold on colony of .355 bees per day. If you lose a third 

### 3 of the bees per day then the colony will collapse but that may 

### 4 actually be partially dependent on what the reproductive rate of 

### 5 the queen is in that colony. 

### 6 So that would be an example of a type of 

### 7 relationships we are talking about. 

### 8 **DR. THOMAS POTTER:**In direct response to the 

### 9 question I'll say that I feel in the case of exposure modeling 

### 10 and for pesticides the state of the science is not well 

### 11 advanced. 

### 12 **DR. DAVID TARPY:**I have no additional 

### 13 comments. 

### 14 **DR. DANIEL SCHLENK:**Other panel members. 

### 15 **MR. JENS PISTORIUS:**I also have this feeling 

### 16 that they are valuable to be considered but that they are not 

### 17 well enough advanced to really be used in the risk assessment. 

### 18 They may inform the risk assessment very well 

### 19 but for the available models I think all have still some lacks 

### 20 which really make potentially use very valuable for this moment. 

### 21 Especially the valuation I think is missing and I think that it 

### 22 has been said for some questions before that the honey bee 

### 23 colonies, they are a very complex organisms and there are so 

### 24 many different factors that I think this will always limit the 

### 25 possibility to describe everything that happens with the bee 

### 26 colony.

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1 Nevertheless, I think there is some use but I 

### 2 also know there have been recent publications and also for the 

### 3 query model in recent discussions we did identified that there 

### 4 is one important factor missing but at the moment I cannot 

### 5 recall which one it was exactly, but I can write that into the 

### 6 notes too. 

### 7 So I think at the moment the models that we 

### 8 have are useable to inform risk assessment but again like with 

### 9 the comment on the question before, I think the most important 

### 10 is that we still rely on the higher tier test. 

### 11 **DR. DANIEL SCHLENK:**Other comment. No one 

### 12 else? Okay. Mr. Sappington do you have any questions of 

### 13 clarification? 

### 14 **MR. KEITH SAPPINGTON:**I think we are good, 

### 15 thank you. 

### 16 **DR. DANIEL SCHLENK:**All right, we are on our 

### 17 last question, I think, perhaps, maybe. You want to read (c) 

### 18 into the record please? 

### 19 **MR. KEITH SAPPINGTON:**Question 14, final 

### 20 subpart; please comment on the important elements that should be 

### 21 considered in reviewing available honey bee colonies, ecological 

### 22 models for potential application in risk assessments. 

### 23 **DR. NINA FEFFERMAN:**The first two questions 

### 24 that should be asked about any model are what insight does it 

### 25 provide that accurate empirical studies could not, or if there 

### 26 are none, how does the model spare efforts or expense to provide

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1 those insight? 

### 2 In the case of risk assessment there are two 

### 3 main components, and Dr. Potter was specifically addressing this 

### 4 earlier as well; how likely is the scenario in question to occur 

### 5 and how severe is the likely outcome once that scenario has 

### 6 occurred. 

### 7 In the case of colony-level ecological models 

### 8 the former question relates really to sort of a dose response 

### 9 aspect of do we get doses out there and if the dose is out there 

### 10 does it get on a bee? 

### 11 Where models actually help with that is kind 

### 12 of questionable because if we have the understanding about those 

### 13 routes of exposure, we can do a model that follows those but the 

### 14 routes are either basically there or not and its impact is 

### 15 either there or not. And we can have some sub-position models 

### 16 but, nah. 

### 17 The latter question relates to how does those 

### 18 individual affects actually both interact together in one bee 

### 19 and how does that scale up into a society of bees and how is 

### 20 that society of bees scale to a population of bees and how does 

### 21 those population of bees provide the conservation and service 

### 22 endpoints that we have been discussing? 

### 23 Those questions are a sweet spot for the 

### 24 ability of models. These relate to the impact of individual 

### 25 responses at sub-lethal effects. They can be ways to study the 

### 26 concomitant effects of different pathways. And we were talking

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1 about I think you guys call them brews of pesticide before or 

### 2 something like that - mixtures, unintended or un-marketed 

### 3 mixtures of pesticide. This is a great place to try and explore 

### 4 those things. 

### 5 For risk assessment itself and the models here 

### 6 presented that were described in the White Paper - and if I fail 

### 7 to mention it I'm sorry - I agree the review of the simulation 

### 8 models in the White Paper was wonderfully done. And as you guys 

### 9 said not completely exhaustive but representative and well 

### 10 described so thank you for doing that, that made my comments 

### 11 able to back up to say and analytic instead of just, oh do not 

### 12 do that with simulation. 

### 13 So as those model presented were not 

### 14 specifically designed with these types of questions in mind, 

### 15 they do not specifically address these particular questions of 

### 16 risk assessment directly and in useful ways; but they could be 

### 17 very important in formulating answers within each level of 

### 18 concern to produce a globally useful risk assessment. So this 

### 19 is where that modularity that we were talking about earlier 

### 20 comes into play. 

### 21 Well how do we understand the impact of this 

### 22 kind of toxicity to queen fecundity? Or to the dispersion of 

### 23 queen (inaudible) in a colony or to the ability of foragers to 

### 24 get home? Those are specific questions and specific effects 

### 25 that could be plug modularly into an overall model. 

### 26 I would like to reiterate again that I do not

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1 mean to suggest that we should plug in every available module 

### 2 because then both analytic and simulation models become unwieldy 

### 3 and the interpretation is very very difficult. 

### 4 Again I will point to the analytic side of 

### 5 things to both designate and integrate which of these specific 

### 6 modules will be of most impact to the risk assessment outcomes 

### 7 for particular questions. 

### 8 And again by employing those more algebraic 

### 9 analytic models it is going to be easier to assess which of the 

### 10 more specific fine-grain models or simulations require then 

### 11 their own empirical metrics to parameterize correctly and how to 

### 12 validate those by as I was saying sort of incidental validation 

### 13 portions of models. 

### 14 I can give an example but I am going to leave 

### 15 it to Dr. James and others to call me on it if an example feels 

### 16 necessary for this question. 

### 17 These sorts of models both analytic and 

### 18 simulation can generate insight into which valid threshold of 

### 19 outcome for risk assessment are going to be meaningful 

### 20 measureable endpoints for compromising the goals of all of the 

### 21 services we are talking about, both to ourselves and to bees 

### 22 themselves. 

### 23 **DR. ROSALIND JAMES:**So the question asks 

### 24 which of the most important elements to consider in a model and 

### 25 I will add to what was said already. My view might be a little 

### 26 bit different than yours but for both theoretical and simulation

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1 models there needs to be some sort of empirical validation 

### 2 before you use them. 

### 3 And ideally I think for simulation models 

### 4 there really needs to have been done, or you can do it, some 

### 5 sort of sensitivity analysis, meaning that each component and 

### 6 how sensitive is that component to the outcome of the model. 

### 7 And then that tells you something about how accurate the data, 

### 8 how precise the data has to be that are the inputs. 

### 9 And then of course hopefully the model gives 

### 10 you the critical elements that you are looking for as an 

### 11 endpoint, which I would think one of those would be to help 

### 12 identify what endpoints you need to request data from the 

### 13 registrant for. 

### 14 **DR. THOMAS POTTER:**I generally agree with 

### 15 what Dr. Fefferman and also Dr. James has stated, I just simply 

### 16 want to add that there is perhaps another question and this is 

### 17 in direct response to Dr. Fefferman that we would ask about a 

### 18 model. 

### 19 And that is, and again there are certain 

### 20 conditions that you have to apply to this, once calibrated 

### 21 and/or validated can it be used in the absence of additional 

### 22 empirical data to inform decisions. And certainly that would be 

### 23 critically important to the agency when we look particularly at 

### 24 Tier II and Tier III studies recognizing they are expensive and 

### 25 that there is a limited number of studies that can be done. 

### 26 Is it possible to use data from a study that

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1 was say conducted in Illinois to inform decisions about whether 

### 2 or not it is suitable to use the active ingredient in the same 

### 3 way in the state of Georgia for example? 

### 4 So certainly these are critical questions that 

### 5 one would ask about a model in terms of what it can do and how 

### 6 effective it is as a tool because I see it as part of a toolkit 

### 7 that we use in risk assessment rather than maybe the other way 

### 8 around, which is the model is used to simply evaluate risk. 

### 9 And I will sum up by saying and again maybe 

### 10 because I'm sitting in the exposure side of the room here I 

### 11 strongly think that considerably more work can be done and 

### 12 substantial progress can be made on the exposure side, which I 

### 13 think will serve to support particularly the Tier I testing and 

### 14 advance the process. 

### 15 **DR. DAVID TARPY:**I echo the comments by the 

### 16 previous discussants. 

### 17 **DR. DANIEL SCHLENK:**Other panel members. 

### 18 **MR. JENS PISTORIUS:**As the last addition I 

### 19 concur to all what has been said. When mentioning to the model 

### 20 of Henry (phonetic) and specifically because that has caused a 

### 21 lot of trouble and concern and it was a comment I made to the 

### 22 previous answer. 

### 23 I just wanted to point out that was not my 

### 24 work who found that this model in specific had was not validated 

### 25 for the purpose because there was one parameter and maximum 

### 26 daily rate of production of new work, which was not empirically

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1 based. 

### 2 And this is basically the work of Dr. Creswell 

### 3 (phonetic), and he made a technical letter to science, I do not 

### 4 know if it is published yet or not but I have it; it is 

### 5 confidential and it is his work. I want to point this out. 

### 6 And this leads me to my part of the answer, I 

### 7 think considering the high variability, I am not quite sure if 

### 8 it is the same in the U.S., but in European region for instance 

### 9 for colonies to develop it is just important to point out that 

### 10 all of these parameters that are measured and that come into 

### 11 those models actually suits also the geographical location and 

### 12 the climate conditions. 

### 13 For instance, colony growth is a completely 

### 14 different issue. When you have southern Europe, you have 

### 15 basically the colony growth is like this. There is a higher 

### 16 population in winter and it grows slower in spring and does not 

### 17 achieve such a growth rate basically. And then it goes to a 

### 18 moderate amount of bees. But what they do in Germany is they 

### 19 come from a very small number amount of bees and then there is 

### 20 basically explosion of the colony. You can call it a really 

### 21 massive increase of number of bees. And then in autumn there is 

### 22 a very big decrease. 

### 23 And when those models are suitably adapted, I 

### 24 am very happy. 

### 25 **DR. NINA FEFFERMAN:**I want to thank Mr. 

### 26 Pistorius for providing a fantastic example of what an analytic

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1 model can show you, what kinds of colony growth trajectories you 

### 2 could expect, what are the parameters to change without having 

### 3 to go measure all of the parameters. Awesome, thank you. 

### 4 **DR. DANIEL SCHLENK:**Any other comments? 

### 5 **DR. ROSALIND JAMES:**I will add one more that 

### 6 I forgot to say. On the review of models you have the de grande 

### 7 (phonetic) bee pop model, but the newer varroa pop model is 

### 8 probably better and includes climate data in it also. 

### 9 **DR. JEFF PETTIS:**This was a follow up to 

### 10 that. The only problem with using simulation models solely for 

### 11 that is that a northern researcher would never get to work in a 

### 12 southern climate, so that would be a drawback. 

### 13 **DR. NINA FEFFERMAN:**For simulation I agree. 

### 14 For analytic, um. 

### 15 **DR. DANIEL SCHLENK:**Any other comments from 

### 16 the panel? Okay, let me go to the agency; any questions of 

### 17 clarification? 

### 18 **MR. KEITH SAPPINGTON:**I think we are good, 

### 19 thank you. 

### 20 **MR. STEVEN BRADBURY:**I think it compliments 

### 21 what you all are talking about to the extent it can get captured 

### 22 in the report could be helpful. 

### 23 So in my mind the Tier II or Tier III studies 

### 24 are physical models trying to provide insights into the real 

### 25 world, whatever the real world is. But you can only do a Tier 

### 26 II or Tier III study in certain places at a certain point in

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1 time with that batch of bees. And either implicitly or 

### 2 explicitly we are running a model in our brain to project what 

### 3 that means in many places and at many times with different races 

### 4 of bees or whatever. And so those empirical models have 

### 5 shortcomings. I mean in empirical like those physical models. 

### 6 7 But all of those things are rolling through 

### 8 our heads and then the simulation models or the analytic models 

### 9 are ways to try to think about all of those things rolling 

### 10 around in our heads and using mathematics and other kinds of 

### 11 knowledge to try to help us think through that. 

### 12 And none of them are right but all of them 

### 13 give us different insight and they are all at various stages of 

### 14 development. 

### 15 So to the extent that question 13 and 14 

### 16 emerge in your writing, if there is any way that you can opine 

### 17 or provide advice on how you try to blend what we know today 

### 18 between insight of simulation or an analytic model might provide 

### 19 to help us be more explicit in the assumptions that we are going 

### 20 to use when we look at those physical models to make projections 

### 21 about what is happening in the southeast or what is happening in 

### 22 the northwest of the United States or the parries in Canada, the 

### 23 Sacramento watershed in California. 

### 24 Because there is probably a blending of all of 

### 25 these, those physical models and what we do not really get real 

### 26 clear about what we are assuming when we look at the results of

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1 physical models and how these analytical or simulation models 

### 2 can help inform what we should be saying about our assumptions. 

### 3 I think that would just be helpful if 

### 4 something between 13 and 14 could try to blend what I think I 

### 5 heard in a lot of the comments but if you guys could bring it 

### 6 together that would, I think, help a lot of people not just us. 

### 7 Thanks. 

### 8 **DR. DANIEL SCHLENK:**Okay, at this point this 

### 9 will be our final sort of commenting period so I would like to 

### 10 just go around the panel, if you have any final comments you 

### 11 would like to get on the record this is the time to do so. And 

### 12 begin with Dr. Sandy. 

### 13 **DR. MARTHA SANDY:**I do not have anything to 

### 14 add. 

### 15 **DR. STEPHEN KLAINE:**I for one would just like 

### 16 to thank the EPA for the document; it was great, and my bee 

### 17 colleagues for educating me about bees. This is a really 

### 18 important and very fascinating subject and I really look forward 

### 19 to see how this thing emerges. It has become obvious to me that 

### 20 we are not dealing with single bees to protect but we are 

### 21 dealing with colonies of bees, at least for honey bees. 

### 22 So I think that is going to require some sort 

### 23 of out of the box thinking as we move forward. Concepts like 

### 24 homeostasis might be better applied to bee colonies than to 

### 25 bees. So anyways I just think it has been an excellent week and 

### 26 very informative.

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1 **DR. MARTHA SANDY:**If I could have a go back I 

### 2 would also like to say I really enjoyed meeting all of you and 

### 3 enjoyed the White Paper too to the EPA staff, thank you. 

### 4 **DR. JAMES MCMANAMAN:**I echo Dr. Sandy and Dr. 

### 5 Klaine's comments and appreciation of the White Paper and all 

### 6 the work that has gone into this and the discussions of the 

### 7 panel. 

### 8 I think the one thing that I came away from 

### 9 this was that, and it emphasizes that I do not think -- I think 

### 10 it was a clear indication that survival is not the only endpoint 

### 11 that should be considered and that should be taken to heart by 

### 12 the agency in terms of looking at other ways of evaluating the 

### 13 effects of pesticides. 

### 14 **DR. KENNETH DELCLOS:**I will just say it was a 

### 15 most interesting week but I have nothing to add here. 

### 16 **DR. DAVID TARPY:**I would like to echo all of 

### 17 those same comments. I think this has been a fantastic 

### 18 discussion, one that has been needed and I look forward to the 

### 19 evolution of this process. 

### 20 I hope everybody has gained an appreciation 

### 21 for just how messy and dirty honey bee colonies can be. And I 

### 22 think that that does not distract at all from their importance 

### 23 to our commercial production agriculture and just what an 

### 24 important issue this really is for our entire system. 

### 25 **DR. PAUL SCHWAB:**This has been quite an 

### 26 education for me. I did not have very much information about

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1 bees before I started reading this White Paper. So my learning 

### 2 curve has been very steep but I appreciate the patience that all 

### 3 of you have express when I have been inclined to weave my way 

### 4 into this panel. 

### 5 Just a little bit of bookkeeping, I did send - 

### 6 I have talked to Christina about this and to Tom - but I did 

### 7 send in a comment dealing with a correction to the absorption 

### 8 equation that was being used for contamination of puddles during 

### 9 a spraying event. 

### 10 I know that we were supposed to have all of 

### 11 this into the record or it really could not officially exist so 

### 12 that is the point of this. 

### 13 **DR. THOMAS POTTER:**I would like to thank 

### 14 first off the EPA staff that put together the White Paper, 

### 15 obviously a huge amount of work and probably done on a 

### 16 compressed timeframe I would expect. It was certainly 

### 17 informative and I can say I learned a tremendous amount from 

### 18 reviewing it and really enjoyed being part of the panel. 

### 19 And I will say that I think that the outcome 

### 20 of this is certainly going to have a very strong impact, I 

### 21 think, on the whole process of ecological risk assessment. I 

### 22 think there are many core elements of this that will transcend 

### 23 into the whole area of looking at ecological risk. 

### 24 **MR. JENS PISTORIUS:**First one technical 

### 25 comment to the bee issues we are talking about before and then 

### 26 on to the rest. I have been making a list with all the points,

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1 you know, where I thought during the discussions I hoped this 

### 2 was going to be mentioned. 

### 3 One last comment I have for synergistic 

### 4 effects for spray application on flowers and crops; when you 

### 5 know that there are a certain -- and I know that it is taken 

### 6 into consideration, I just want to point it out again -- that 

### 7 there are synergistic actions between two classes of compounds 

### 8 like the pyrethroids and the EBI fungicides. Please consider 

### 9 those synergisms as well as all others really documented. We do 

### 10 not have to test every sector, but with those two classes 

### 11 pyrethroids and EBI fungicides. 

### 12 Please also do a higher tier risk assessment 

### 13 or do risk mitigation measures that are appropriate because we 

### 14 know we had problems with it in the past. But I think this is 

### 15 taken into consideration so I'm going to end it with a technical 

### 16 comment. 

### 17 But now let me say a few personal words on 

### 18 this whole issue. Before I met the first EPA staff in 2011 at 

### 19 the SETAC Pellston conference, I will tell you frankly that one 

### 20 person told me what is going to happen. You are going to the 

### 21 U.S. and they'll let you talk, let you know your experiences and 

### 22 then they will say, thank you, and not take anything of this 

### 23 into account. 

### 24 And I am deeply impressed, seriously, deeply 

### 25 impressed and I will go to this person and I will tell him all 

### 26 of my experiences. Really it is such a great honor to be here.

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1 I was really impressed by all the good work. I have to thank 

### 2 the whole EPA staff. I think it is such an incredible effort 

### 3 that you have made. 

### 4 This White Paper, when I read it I thought, 

### 5 hey this is great. Really, I think there is so much effort in 

### 6 this and I must say really I am deeply impressed how far you 

### 7 have brought this and how good the advancement is and we have 

### 8 been working together a little bit on other issues since 2011; I 

### 9 love to work with you. And it has been great fun, great honor 

### 10 and I think this has been such a great scientific progress. 

### 11 Thanks a lot, I take my hat. 

### 12 **DR. JEFF PETTIS:**I would just like to offer 

### 13 thanks for the invitation to be part of the panel. I have very 

### 14 much enjoyed it. I would like to thank EPA for outlining the 

### 15 three protection goals, in particular, for me protection goal 

### 16 three which was broader than just honey bees. We tend to get 

### 17 caught up in honey bee biology and the complexities of that, but 

### 18 pollinated declined is bigger than just honey bee issues. 

### 19 So I thank the effort put into the White 

### 20 Paper, in particular the protection goals I thought were spot 

### 21 on. Thank you. 

### 22 **DR. NANCY OSTIGUY:**This was probably the most 

### 23 intense, but also the most fun four days that I have spent in a 

### 24 long time. Everyone has been incredibly collegial. It has been 

### 25 very informative. I have enjoyed myself thoroughly. I have 

### 26 collected also an amazing amount of information I am going to

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1 take back to my students. 

### 2 I have always been a defender of EPA because I 

### 3 think you get picked on too much but I have some very personal 

### 4 stories now to tell my students about what a wonderful job you 

### 5 are doing under very difficult circumstances. 

### 6 And I also have to say something in defense of 

### 7 those messy bees. They are no messier to think about than when 

### 8 you think about human as a population. I feel extremely sorry 

### 9 for the sociologists that are trying to study us. And in many 

### 10 ways maybe we should think of ourselves as sociologists of honey 

### 11 bees. 

### 12 But in defense of those bees they are 

### 13 extremely neat inside those colonies. 

### 14 **DR. ROSALIND JAMES:**I almost want to say, I 

### 15 concur with my colleagues because I guess I will say the same 

### 16 thing. I was very glad to see EPA take on this issue. Everyone 

### 17 has been complimentary; I do not want to be too uncomplimentary 

### 18 but I do want to say, finally. It is a little behind times but 

### 19 I am very glad to see EPA take it on. 

### 20 And when I started reading the White Paper I 

### 21 was very impressed. I mean throughout it. It was better than I 

### 22 expected I guess is what I mean. And I appreciate being invited 

### 23 to this panel with my beyond Apis mellifera views. Thank you. 

### 24 **DR. GREG HUNT:**I concur; and I also am having 

### 25 my students read some papers that relates to these issues and we 

### 26 will be talking about it next week.

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1 And I thank the EPA for their efforts. I 

### 2 think that the increase emphasis in Tier I that is put in the 

### 3 White Paper is very good and I hope that we as a panel can 

### 4 provide a kind of a tiered response in our writings so that we 

### 5 can give you some, putting the simple things at the top and then 

### 6 the more future things at the bottom. 

### 7 **DR. NINA FEFFERMAN:**Thank you guys, all of 

### 8 you, the EPA members, the panel the public commenters, you guys 

### 9 running this, oh my God. 

### 10 I have had an awesome time and I have learned 

### 11 a huge amount and I have made friends and I like it. As I read 

### 12 into the existence record I want to say that there are some 

### 13 analytic models of honey bee colonies and I will provide some of 

### 14 those for the notes. Thank you. 

### 15 **DR. MAY BERENBAUM:**And to come crashing down 

### 16 on an incredibly mundane technical note just to read it into the 

### 17 record but I will follow that with some more inspirational 

### 18 language. 

### 19 Our discussions of prolonging the endpoint of 

### 20 Tier I larvae assay through pupation lacked literature 

### 21 documentation. There were some hand waving or remarks about, 

### 22 yes papers are out there. Well one such paper is "Honey Bee 

### 23 Risk Assessment: New Approaches for In Vitro larvae Rearing and 

### 24 Data Analyses" by Hendriksma, et al. "Methods in Ecology and 

### 25 Evolution" 2011. It is sort of a modification of Aupinel et al, 

### 26 but the point is it is a non-grafting method and the endpoint is

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1 pre-pupation and it was used with some success. 

### 2 On a higher note, as I mentioned 117 years 

### 3 inadvertent bee mortality caused by pesticide is really -- I am 

### 4 deliriously happy that it is now a national priority to protect 

### 5 not just honey bees but bees in general and I do not know if 

### 6 people will appreciate what a massive societal change that is. 

### 7 Insect toxicology for decades involved how to 

### 8 kill insects so this is really un-trodden ground for the most 

### 9 part. And having said that the repeated references to data 

### 10 gaps, I think, obscures the fact that the White Paper represent 

### 11 there has been a tremendous explosion of extremely informative 

### 12 and relevant data so that we now can make science-based 

### 13 decisions on the best way to access risk and protect bees. 

### 14 So I am glad bees are having their day here in 

### 15 Washington D.C. and thank you for letting me be part of it. 

### 16 **DR. DANIEL SCHLENK:**Let me say you are all 

### 17 making history. As I mentioned before this was a record number 

### 18 of questions that we have ever gotten, and you did it. You did 

### 19 do it. I want to congratulate you. This has been one of the 

### 20 best panels I have had the honor of being a part of. 

### 21 I appreciate the tremendous hard work that 

### 22 many of you did throughout this week and days leading up to it 

### 23 of course. But also just to your collegiality and working 

### 24 together and being efficient in terms of answering the 

### 25 questions; that was just phenomenal. Great job, hat off to you 

### 26 guys.

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1 **DR. JEFF PETTIS:**A point about your 

### 2 leadership on the panel. There were some issues when you called 

### 3 it yesterday at 4:30, and we were like "why not press on?" and 

### 4 so there was a lot of discussion afterward but you made the 

### 5 right call there. We never would have gotten through these last 

### 6 questions with the level of detail and discussion that was 

### 7 needed. So you made a great call yesterday. We were pretty 

### 8 burnt yesterday. 

### 9 **DR. DANIEL SCHLENK:**Thanks, actually that was 

### 10 Dr. Jenkins pushing me on the right side over there. I wish I 

### 11 could take credit for that but. 

### 12 **DR. NINA FEFFERMAN:**I also just quickly want 

### 13 to acknowledge the amazing staff that has been helping us 

### 14 throughout; they have made this run smoothly as well and I want 

### 15 to thank them. 

### 16 **DR. DANIEL SCHLENK:**Definitely, where would 

### 17 we be without the SAP staff? Thank you very much, getting you 

### 18 all here and out safely and soundly hopefully. 

### 19 Again, thanks to the panel members for your 

### 20 tremendous work. It has been a pleasure to be a part of this. 

### 21 Obviously the EPA for the presentation, I totally concur on the 

### 22 White Paper and one of the things I think you hear at least 

### 23 among the academics is, even though you never what happens in 

### 24 terms of the policies that come out of this but in terms of 

### 25 academia and education the things that you are doing are making 

### 26 a difference at least in student's lives. Because I know every

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1 single academic in here uses this information to educate our 

### 2 students. And that to me has always been a tremendous benefit 

### 3 of being part of these things; it is just so educational not 

### 4 just for me but for people that we also interact with. 

### 5 So it is an exponential type of information 

### 6 sharing that comes out of this. So just congratulates you on 

### 7 that and again, I thought the White Paper was amazing. Not 

### 8 being familiar with this particular topic and the detail and 

### 9 clarity that was addressing these particular issues was just 

### 10 fabulous, kudos for that as well. 

### 11 I would like to thank our public commenters, 

### 12 if they are still here, probably all gone but for their input 

### 13 and realism in terms of what this issue really means to a lot of 

### 14 those people. 

### 15 EPA staff as well; I already said that. And 

### 16 with that I will turn it over to Dr. Bradbury usually have a 

### 17 word or two to say at the end. 

### 18 **DR. STEVEN BRADBURY:**Real quick, just want to 

### 19 thank again the panel for all the hard work. We know we had a 

### 20 record setting number of questions and I appreciate all the work 

### 21 to do it. And every single topic you guys worked through I 

### 22 think gave us a lot of valuable insights. And the way you were 

### 23 synthesizing information like a super colony in action so we 

### 24 appreciate that too. 

### 25 And we are committed to moving forward and I 

### 26 appreciate the comments about incremental steps and feedback

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1 will help us move bit by bit and we will try to do better and 

### 2 better with our colleagues in Canada and California and around 

### 3 the world that we are all working on this. 

### 4 I just want to thank you; you are a big 

### 5 contributor to advancing science and our ability to evaluate 

### 6 these situations. 

### 7 **DR. FRED JENKINS:**I first want to start off 

### 8 by thanking Dr. Schlenk; this was a very important meeting, 

### 9 obviously a record breaking meeting and he did a great job in 

### 10 leading this panel through these charge questions. 

### 11 And thank you so much to this panel. You all 

### 12 were fantastic and I appreciate you all agreeing to serve on 

### 13 this panel. When I send my invitation, I find you, send my 

### 14 invitations and talk to you and I really appreciate you all 

### 15 expertise, your talents, your commitments in this effort. And 

### 16 it has been truly a pleasure this whole week working with you 

### 17 all. 

### 18 To the EPA-OPP team, there is a lot of hard 

### 19 work that goes on building up to having this meeting and OPP 

### 20 really works very hard. I used to work in OPP so I know and I 

### 21 used to work with EFED as a biologist, as an eco risk assessor 

### 22 for a long time. And so I know the work ethics in that office 

### 23 and I appreciate the opportunity to be there working with you 

### 24 all for this meeting. 

### 25 Special thanks to Dr. Steven Bradbury the 

### 26 leader of the Office of Pesticide Programs. Dr. Donald Brady,

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1 EFED Director, and the Branch Chief who actually led the team 

### 2 who did this work. Dr. Thomas Steeger, Thomas Moriarty, Mr. 

### 3 Keith Sappington, Christina Wendel, Kristina Garber, Reuben 

### 4 Baris, Joseph DeCant, all the speakers and scientists involved 

### 5 in developing this very good White Paper. 

### 6 Thank you for the collaborative efforts of Cal 

### 7 EPA and PMRA, Ms. Mary Mitchell and Richard Bireley from Cal EPA 

### 8 Department of Pesticide Regulation. Thank you for your efforts 

### 9 in this whole process. 

### 10 Thank you to all the public commenters and to 

### 11 all those who provided written public comments; there are a lot 

### 12 of written public comments in regualations.gov; so thank you to 

### 13 all those in the public for participating in this process. 

### 14 And I would be of remiss if I personally did 

### 15 not thank my FIFRA SAP colleagues under the leadership of Laura 

### 16 Bailey, my colleagues Joseph Bailey, Sharlene Matten, Joyce 

### 17 Coates, Lu-Ann Kleibacker, our team lead, and Shirley Percival. 

### 18 19 The meeting report will be available no later 

### 20 than 90 days after the close of this meeting. This meeting is 

### 21 now officially concluded. Everyone have a nice weekend and 

### 22 thank you all. 

### 23 (WHEREUPON the meeting was adjourned) 

### 24 25 26

### FIFRA SCIENTIFIC ADVISORY PANEL (SAP)

**OPEN MEETING**

**POLLINATOR RISK ASSESSMENT FRAMEWORK**

**DOCKET NUMBER: EPA-HQ-OPP-2012**-**0543 FIFRA SAP WEB SITE http://www.epa.gov/scipoly/sap/**  **OPP Docket Telephone: (703) 305-5805**

**UNITED STATES ENVIRONMENTAL**

**PROTECTION AGENCY**

**CONFERENCE CENTER LOBBY LEVEL ONE POTOMAC YARD (SOUTH BUILDING)**  **2777 SOUTH CRYSTAL DRIVE ARLINGTON, VIRGINIA 22202**  **SEPTEMBER 11 TH** 

### - 14**TH**

### , 2012

### FIFRA SCIENTIFIC ADVISORY PANEL DOCKET #EPA-HQ-OPP-2012-0543

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17:11 156:9 158:8 200:17 277:24 278:2,6,9,14 279:1 280:7 292:21 347:16 684:22 695:2 Canada's 8:9 15:2 17:2 292:13 293:2 Canadian 15:7 280:14 292:23 311:18 canals 82:12 cancelled 324:13 cancer 335:19 492:25,26 candid 150:21 candidates 509:22 cannibalism 226:6 canola 148:22 600:19,19,23 602:25 CAP592:19 capabilities 12:23 235:5 291:22 capability 246:1 capable 558:17 capacity 133:20 447:25 458:13 502:8 645:18 capillary 537:23 capped 304:2 342:9 534:14 capping 346:21 456:17 caps 595:15 Captan 305:16 308:14 capture 25:5 67:7 136:12 155:18 222:7 352:26

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347:12 426:9 447:17 647:3 683:21 captures 12:11 647:6 capturing 67:8 404:6 420:13,14 537:22 carbamate 120:10 398:26 carbamates 373:22 398:19 carbofuran 383:5 carbohydrates 467:1 carbon 537:22 carcinogens 492:26 care 232:7 241:24 334:5 484:26 489:19,26 490:3

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461:12 486:21 540:18 588:7 595:17 657:2,19 CFR 17:6 156:6 167:20 168:2 chain 313:21 Chair 3:21,21 13:3 249:14 644:3 Chairman 18:3 441:26 643:11 chairs 251:21 challenge 6:22 10:11,14,14 51:8 163:24 189:22

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43:19 55:10 58:1,5,10,10 65:2 70:26 72:7,23,24 74:3,4,6,24,25 75:14,26 76:9,11,12,18,24 80:21 80:24 81:2,14 82:1 89:26 91:24 92:1,21 106:10 106:11 108:13,17,18 110:4,10 111:2,11,14,24 112:16 113:2 120:6,16 126:23 128:25 129:6 130:20 135:18 136:23 137:2 156:7,14,18,25 157:7,16,23 158:6,15 159:1 167:13 207:5,8,21,23 208:10 215:19 216:10,22,25 217:2,25 218:2,14 218:17 257:14 267:6 289:4 307:15 315:25 326:13 326:15,18 345:26 346:5,8 347:17 351:8 355:8 362:1,6 365:23 366:9 371:15,17 373:8 375:7 407:24,26 408:10,24,25,25 411:4 413:17,24 414:26 422:24 429:10 432:23 433:5,7,9 434:11 434:14,26 438:6,13 441:1 467:13,17 474:5 476:24 506:10 518:21 520:3 521:14,16,20 540:24 602:1 603:20,22 606:15 607:11 612:11 618:18 632:9 662:13,20 contacts 181:20 434:22 contain 55:12 64:25 86:14 99:24 141:26 191:11

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69:6 90:12,17,17 92:8 93:26 107:21 119:21 121:9 126:6 129:24 140:20,25 141:13,15,18 142:9,12 143:16 144:6 146:3,6 163:5 164:19 165:4,8 171:12 179:14 198:14,16 201:3 202:13,16 203:15 204:20 205:1,1,2,3,12,21,24 206:8 208:23 209:10 209:19 210:25 219:4 240:6 258:5 259:23 260:23 260:25,25 261:9 367:14 383:22 396:5,12 397:11 398:14 403:21 404:1 405:21 420:19 426:5 430:16 434:12 466:14 491:21 500:3 603:12 609:2 estimate-based 205:7,11 estimating 88:15 89:2,26 90:2 106:9,16 108:12,13 108:16 122:26 125:21 126:15 128:5 130:20 205:15 207:4 208:13 273:18 374:16 388:4 397:19 405:21 426:17 434:13,25 484:17 492:2 555:17 612:11 estimation 25:22 91:20 120:22 204:24,26 206:18

208:21 209:9,15 497:23 633:7 et 19:8 96:7,8,13 97:3,6,14 101:24 107:7 113:16 117:11 120:8,20 125:26,26 126:3 159:15 171:1 196:26 224:26 225:8,18,25 227:2 237:8 259:22 262:19 298:11,13,16 299:12 319:8 324:24 329:22 332:4 387:18 398:1 431:18 433:8 443:23 461:12 464:25 472:7 475:14 486:21 500:3 540:18 588:7 595:17 636:23 657:2,19 674:26 691:24,25 ethics 2:21,26 3:2 695:22 EU 161:7 273:16 380:26 381:9 390:11 Europe 19:7 273:7,7,20 482:23 592:6 610:26 628:3

682:14 European 11:2 22:25 23:20 24:16 29:13 64:11 101:2,5 105:1 107:4 137:5 156:21 158:9 202:24 213:15 225:4 228:14 491:3 660:15 665:14 682:8 evaluate 18:16 22:17 27:20 48:14 108:7,11 111:26 146:22 149:3 156:4,24 157:1,3,6 158:4 160:12 161:3 163:17 170:23 173:15,16,25 175:17 178:1 178:7,9 179:13 185:14,15 194:14 209:2 225:19 244:18 393:7 433:10 446:17 461:11 565:3 662:6 681:8 695:5 evaluated 54:20 57:14,25 58:12 60:14 61:15 62:4,7

62:18 76:22 80:23 107:19 117:26 155:5,7 165:7 177:19 204:20 214:4 217:13 219:2 228:26 240:12 670:24 evaluates 280:26 evaluating 13:17 24:3,14 31:5 59:10,11 61:19 67:2

90:16 128:13 144:22 154:12 155:13,23 164:2 167:10 201:2 208:17 211:19 212:11,18,23 213:7 220:18 283:12 300:23 346:12 455:8 585:24 638:14 656:19 686:12 evaluation 15:20 27:24 49:21 55:16 58:25 61:17 107:12 108:8 111:24 112:4 113:22 119:1 154:18 155:25 159:16 161:24 184:4 192:9 212:3,10 217:8,14 276:2 347:18 406:26 517:26 590:22 evaluations 155:26 168:26 Evans443:22 evaporate46:11 428:3,4 evaporated46:1 429:26 439:25

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www.transcriptionetc.com Page 44 evaporates 429:26 evaporation439:24 evaporative 38:21 77:24 evasive 291:10 evening 325:6 435:22 evenly 259:6 event 15:5 17:13 238:8,11 557:1 687:9 events 35:14 163:2 287:16 300:21 557:6 eventual 20:11 eventually 156:4 227:19 509:26 evert511:26 everted 511:21 everybody 249:12 273:20 275:17 296:17 354:25

407:17 436:1 442:1 499:13 513:15 525:24 621:26 624:14 667:16 686:20 everyday485:15 520:22 evidence 25:1,1,24 31:1 48:14 55:24 59:18,22 60:6

60:10 61:11,21 62:6 63:24 66:5,7 153:6,11 158:5 163:12 166:19 167:14 174:25 178:24 181:1,21 194:20 203:15 209:3,10,12 210:21,26 211:3,8,19 211:22 212:3,6,23 213:7,8,10 214:10,15 219:7,16 219:22 229:1 253:25 378:1 455:4 533:15 539:9 542:10 587:14 621:7,14,25 630:23 639:23 evidenced 83:25 evident 145:9 169:2 eviscerated 511:17 evolution 686:19 691:25 evolve 25:11 52:11 204:16 evolves 51:4 evolving 51:19 exacerbate 219:17 exacerbated 527:22 exact 365:12 391:13 520:10 531:18 568:15 594:5

655:18 exactly 232:25 241:19 242:18 307:12 352:3 363:14

391:20 402:25 455:12 461:26 462:6 523:1 546:5 557:11 567:9 582:25 592:10 608:20 610:17,17 650:9 652:8,15 659:24 660:9 670:10 673:6,7 676:5 exaggerate 275:6 exaggerated 171:17 275:10 examination 215:21 examine 213:22 398:17 400:18 examined 23:25 75:22 281:16 398:19 411:7 609:2 examining 21:19 181:5 399:7 623:8 example 11:19 15:10 31:16 35:10 38:11 39:7,12

55:7 56:26 61:25 64:26 65:12 73:23 108:10 119:17 124:1,16,21 128:16 135:4 142:3 147:18 148:20 156:1 162:14,23 177:2 207:12 211:20 218:24 221:26 222:21 225:8 227:1,22 230:16 231:3 240:24 261:23 265:2 277:2 282:6,15 313:15 317:12 344:10 353:9,11 360:8 388:21 411:19 436:24 438:23,26 443:22 465:2,17 516:10 550:21 552:23 555:8,8 560:21 588:5 597:20 620:10 634:5 645:3,10 652:23,24,25 653:1 654:6 655:3 661:12,13,18 666:3 670:5 674:20,25 675:6 679:14,15 681:3 682:26 examples 30:14,20 153:17 216:14 277:3 337:7

560:20 645:1 651:21 663:17,18 665:11,13 exceed 78:26 98:16 115:2,4,8 117:1 119:4,6 141:17

217:17 218:14 291:22 393:14 426:8 exceedance437:2,3 exceeded 67:20 114:19 205:13 400:10 exceedingly 381:5 424:4 656:7 exceeds 67:18 99:14 115:14 123:10 439:1 Excel 647:6 excellent 15:10 197:3 215:1 315:14 331:4 334:3

343:16,18 364:11 440:6 442:3 469:26 571:11 633:16 674:22 685:25 exception 123:16 365:3 369:15 384:25 385:2

520:13 605:25 642:20 exceptionally 657:5 excerpt206:12 excessive 221:26 289:25 excessively 550:24 exclude 327:8 excluded 327:14 351:14 excluding 347:18 exclusion 224:2,22 234:10 exclusively465:5 excreted 265:5 excruciating 242:9 excuse 69:12 231:23 290:17 409:8 572:17 658:17 executed 562:18 executive 202:23 223:17 224:5 exercise 214:23 218:12 294:2 exert487:13 500:2 exhaust 300:7 exhaustive 30:20 224:12 406:15 678:9 exhibit 150:2 298:8 499:21 exist 159:3 161:1 165:24 243:22,22 280:7 343:6

410:2 615:14 616:22 687:11 existence 240:23 475:9 691:12 existing 21:23 56:8 147:4 150:22 227:25 244:8

294:26 301:21 617:13 618:7 657:12 663:13 exists 158:14 172:19 240:25 294:5 641:1 exiting 54:24 exoskeleton 356:3 expand 171:21 expanded 156:23 157:9

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452:17 498:11 527:25 640:9 641:9 655:22 683:2 687:16 expectation 56:10,25 640:12 expectations 195:18 expected 36:1,6 38:26 40:25 42:3 79:6,10 80:3

87:4 94:3,4 116:25,26 118:24 119:10,18 124:2 148:10 157:19 170:9,12,14 217:10,26 218:22 373:17 422:23 429:9 486:10 588:4,19 602:13,14 653:13 655:23 690:22 expecting44:19 expediting 345:8 expended 50:8 expense 155:19 676:26 expensive 363:24 680:24 experience 16:9 152:13 176:10 238:23 263:21

280:16 351:2 380:16 467:9 496:1 541:17 552:6 553:5 554:3 574:24 578:10 591:3 592:18 603:5 622:23,24 638:18 662:10 experienced466:8 experiences 688:21,26 experiencing 238:4 239:2 experiment 172:1 192:1 421:6 521:4 524:19 525:4

535:15 542:21 547:7 571:24 642:1 experimental 177:2 182:22 212:18 343:9 344:14

375:2 392:26 398:1,9,12 399:10,15 458:20 496:20 500:13 527:4 567:10 579:4 596:17 606:6 633:19 experimentally 650:11 experiments 231:21 246:23 418:20 419:2 521:3

524:26 534:17 562:16 598:10 634:2 672:20,22 674:9 expert7:15 23:8 85:14 263:25 638:26 668:14 expertise4:2,5 5:9,25 22:21 290:2 635:4 647:2

651:15 695:15 experts 138:15 242:8 555:9 653:5,13 671:14 explain 242:11 313:26 432:25 644:26 664:8,18,20 explained 194:21 272:9 318:17 664:8 explaining 107:9 164:10 241:15 634:23 660:8 explains 355:18 explanation 256:21 355:17 644:1 explicit29:19 203:24 223:24 225:16 226:14,19

227:11 666:3 684:19 explicitly 59:16 65:24 294:18 684:2 Exploratory 312:19 explore 224:14 511:19 678:3 explored 155:13 171:20 178:4 explosion 682:20 692:11 exponential 101:20 694:5 expose74:21 583:25 exposed 38:23,26 40:11 41:11 72:26 73:18,25 74:8

74:15 81:26 132:4 142:15 158:22 159:3 165:16 181:11 185:26 200:14 265:12 298:5,8,26 299:3 299:25 300:1,4,6,11 301:4 338:15 351:15,20 352:18 355:7,8,13 356:2 360:16 362:4,5 363:2 376:1 423:15 439:21 444:14 450:26 486:11 522:14 539:3 560:12 576:12,23 579:11 584:3 590:3 595:23 602:1 exposing 132:13 481:8 529:2 exposure 18:11 22:5,9,12,16 23:26 24:5 25:1,5,21

25:26 26:21 30:7 31:12,17 35:2,15 36:7,12,20,21 37:1,5,12,13,17,21,24 38:3,5,6,9,13,19 39:2,12 40:8,13,14,22,26 41:3,8,19,25 43:19,20 44:8,20 45:3 46:16 47:2,6,14,18,24,26 54:8,16,17,18 55:4 55:6,7,9,17 57:16,22,24,25 58:5,11,12,13,24,25 59:25 60:25 62:2,19,20,22,23 63:6,10,11,14,16 64:1 65:2,3,5 70:26 71:13,13,15,16,18,22,26,26 72:3,4,6,7,8,9,11,16,19,22 73:3,4,7,7,9,15,17,21 73:26 74:3,4,10,11,14,18,24,25 75:2,2,7,17,18,19 75:20,25 76:1,3,4,5,6,7,9,10,11,14,18,18,20,24 76:26 77:4,16,17,17 78:3,4,6,12,20,24 79:2,3 80:1,6,8,11,15,16,17,17,18,19,21,23,26 81:1,3,3 81:4,5,14,14 82:1,9 83:24 84:1,6 85:2,4,9,17 86:13 87:7 89:2,11,17,17,26 90:11,15,17,19,19 90:21 91:7,11,18,22,26 92:5,8,10,11,12,14,15,18 92:21,22,23 93:23,25,26 95:9,15 98:4,8 100:10 101:8 103:25 104:22 106:9,10,14 107:9,16,24 110:6 111:11,25 113:5 115:20 116:17 117:6 118:19 121:8,9 123:23 126:18,21,23 127:1,19 128:23 129:6 130:21 132:9,11 135:4,18 140:13 140:19,23,25 141:4,5,10,13,15,17,19 142:5,9,12 143:4,7,8,9,15,16,18 144:1,8,10,12,16,23,26 145:5,13 146:2,3,5,10,14 148:6,10,11 150:5,10 151:2,5 155:14,18 156:19,25 157:13,18,22 161:14 164:19 165:3,5,17,19,25 166:3,7,15,19 167:11,13,14 169:10,18,24 170:7 171:22 172:2 172:21,23,25 173:16 174:11,13 175:13,20 177:8 178:25 179:2,6 180:12 183:15,19 185:7,15,23 187:10 189:3,14 190:15 191:12 192:12 193:9,14 193:14 194:10,20 197:22 198:14 199:12 200:11 200:12,13,19 201:2,3,11,24 203:14 204:10,20 205:8,18,22,24 206:2,8 208:14,23,26 209:17 213:1,2,24 216:2,3 217:2 218:2,20 219:1,3 227:26 229:8 237:7 240:6 256:1 257:7,14,21 260:8,23 261:1,23 265:10,15,17 266:19 268:2 275:6,9 291:3,23 292:25 293:16,21 294:25 298:10 300:10,16 301:10 318:3,23 327:1 328:3

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475:12 504:8 expressions 29:20 extant 396:21 extend 2:12 3:19 503:5 524:15 529:15 530:19

537:8 extended 158:16 161:22 176:23 181:5 317:2 428:2

479:20 483:12 529:2 607:8 extending443:13 529:26 530:8 540:7 extension 5:3 250:1 450:20 451:15 460:7 extensive40:19 165:21 185:20 187:6 292:22 428:6 extensively 205:5 504:7 extent 18:20,21 22:16 24:11 31:7,21,25 32:1 42:7 45:13 50:3 70:17 106:2,4 129:10 150:18 167:17 172:25 174:2,14 188:1 189:14 210:8 214:8 226:15 238:20,20,21 258:25 282:9,17,20 283:5 283:13,25 284:19 291:25 294:24 357:18 358:9 361:6 392:14 397:10 403:20 441:21 459:10 464:24 465:4 471:24 472:23 500:19 518:26 520:9 543:8 566:19 568:4,10 576:14 608:3 670:24 683:21 684:15 extra 148:23 242:17 318:8 415:5 435:16 459:16

530:14 589:4 extracted425:2 extrapolate 222:11 277:2 374:9 382:16 387:21

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273:18 284:8 294:4 298:23 300:15 318:1 319:21 356:8 359:24 381:14 388:19 394:12 401:8 418:13 429:21 433:26 466:21 475:14 496:20 555:13 568:7 594:21 595:25 609:12 611:10,16 613:7 617:23 621:8 640:13 644:18 653:26 661:11 670:13 692:10 factor 20:21 104:17 110:9 118:5 119:7 120:13 123:10 188:5 235:26 260:14 265:1 283:4 329:20 330:8,17 377:10 382:9 383:8,11 426:4 438:6 498:13 563:24 570:5 596:18 611:4 631:20 665:20 676:4 factored 84:7 612:23 factors 13:23 19:15,23 20:19,23 21:7 23:5 28:3

39:15 53:21 69:26 83:17 84:1 91:13 166:8 175:19 176:12 177:1,22 190:5 192:22 193:7 211:25 223:4 228:17 233:16 235:1 238:14 239:11 243:10

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268:26 270:7 276:19 279:20 302:21 308:19,24 319:15 325:25 331:23 335:17 344:20 357:2 378:9 411:6 412:15 426:9 433:4 450:6 452:1 482:13 515:15 523:2,18 539:12 544:8 546:11,12 547:12 548:24 550:18 577:7 625:2 626:12 630:19 675:9 690:8 feeling 328:11 347:4 501:4 533:16 546:16 675:15 feels 417:5 667:11 679:15 feet 322:23,23 Fefferman 6:5,5 47:8,9 48:3 49:7,8 50:12 51:20

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239:4 248:1,2 256:24 333:22 337:19 357:19 622:2 meeting 2:3,3,6,13,14,22 3:3,14,21 7:4 21:20

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296:13 303:18 322:24 378:17 379:12 437:26 493:11 546:9 547:17 585:13 643:7 667:14 670:12 miscarriage493:2 mismatch 604:10 misread 610:1 missed 194:25 309:8 607:22 misses 644:9,16 missing 283:6,22 284:2 385:6 510:24 538:9 541:9

674:23 675:21 676:4 misspoke 652:1 mistake 551:23 555:4 560:23 597:16 640:26

650:23 mistakes 262:24 597:20 misunderstand 81:15,16 misunderstanding 242:4 659:3 misuse 630:8 misused 630:7 Mitchell 14:13,25,26 15:1 696:7 mite 231:24 232:11 305:3 637:17 mites 19:25,25 20:1 21:2 231:24 232:8,10 233:6

235:24 237:7 303:22 305:2 306:13 471:6 586:16 637:18,21 miticide471:5 miticides 533:19 mitigate 63:13 238:21 291:11 mitigated 179:19 mitigating 276:15 331:24 414:14 mitigation 60:9,12 61:15,18,20 62:5,7 77:6,8 84:7

84:11,15,19,19 85:4 166:22 179:17 193:23 204:19 276:2,4,8 287:25 331:8,19 410:7 416:18 417:4,6,11 419:11 429:14 525:22 628:26 688:13 mixed 186:15 417:24 mixes 220:13 mixing 307:23 mixture 145:6 159:20 160:8 311:1 421:4 mixtures 300:14,18 678:2,3 Mm-hmm 134:23 mobile 128:12 mobility 119:17,19 124:2,4 298:1 mode 59:19 68:4 170:9 182:26 285:26 412:4

607:17 634:25 635:2 668:15

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37:1 39:20 40:3,23 41:1 42:21 43:2,9,23,25 44:14 44:15,16,18 45:5,16 47:17,21,23 48:5,13 63:5 68:7 71:17,23 83:1 86:2 88:14,15,17 106:25 107:6,8 110:7,16 113:7 120:3,4,18 121:4,18,26 122:1,12 125:21 126:3,11,14 128:6,7 129:13 131:1 132:8 135:16,18 164:19 199:24 202:13,16 206:15 207:11,14,17,26 211:9 216:5 222:9 223:5 223:25 224:1,3,4,7,21,26 225:1,6,6,7,8,12,12,14 225:18,21,24 226:2,3,4,8,23,24,25 227:1,1,6,7,8 227:15,20 228:2,6,14,17,19 229:21,22 231:15,18 231:23,23,25 232:10,11 240:6 241:10,11,12,13 241:14,15,16,17,21,23 243:20 244:3,8,21,22,24 245:1 257:1,3,22 258:18 259:21 260:5 271:20 356:15 366:16 373:25 392:21 393:1 394:1,12 397:11,17,20,25 398:2,5,17 399:3,4,9,14 400:3 401:5,13,25 403:19,19 404:8,14,18,21,26 405:3,4 408:16 411:12 424:12,13,13,15,15 425:1 426:7 426:15 502:9 527:6 550:13 560:24 619:23 644:5 644:22,22 645:7,8,23 647:5,13 648:24 650:2,26 651:1,13,16 653:4 655:17 656:23 657:15 658:8 658:22,23 659:18 660:22 661:15,20 665:5,16 666:14,16,22,23 670:4,5 671:10,22 673:2,4,9,9 673:14,15,17,20,22,26 674:1,5,15,22,24,26 676:3 676:24,26 677:13 678:13,25 679:24 680:6,9,18 681:5,8,19,24 683:1,7,7 684:2,18 modeled411:12 modeler 250:26 modelers 239:18 modeling 6:7 47:2,11 48:17 100:18 199:20 225:17

231:8 239:17,25 242:22 243:5,19 262:22 264:15 264:18 271:16,18 291:2 373:12 398:10 406:9,10 406:16 461:26 500:23 501:9 502:1 511:7 516:24 551:11,17,19 553:4 555:11 606:3,10 627:7 644:7 644:10,18,23 647:25 649:9,13,26 650:24 654:7 654:21 656:16 657:6,13 658:3,5,15 659:4,12 660:5,7,19 662:1,4,8 663:12 664:4,9,14,25 665:12,13,18,23 672:19 675:9 models 27:12 28:7 32:6 33:11,13,18,19,21,24 34:9

34:15,26 35:12,19,22,25 36:4,9,13,17,19 41:24 42:6,12 44:6 49:9,19 55:20 75:20 107:17 125:25 130:21 165:4 199:22 205:25 214:17,18 219:10 221:4,7,7,14,17,20,21 222:14,19 223:1,7,11,16 223:22 224:10,13,17,17,24 226:12,18,20,22,26 227:10,24 228:12,16,23,25,25,26 229:13,17 230:3,3,5,8,8,10,11,11,13,18,19,25 231:2,12 232:18 240:3,4,16 241:3 243:25 244:1,17,19,23 244:23 263:22 271:21 296:26 340:20 345:14,20 345:25 351:10 387:21 401:11 550:9 643:18 644:6 644:26 648:3,13 649:15,22 650:8,19,20,25 651:13 654:22 655:4,19 656:4,19,22 660:11,26 661:10 662:18 663:14 666:6,13 667:8,11,13,17 670:21,24 671:3,4,4,11 672:3,7,13,14,18,18,19 672:20,23 673:6 674:23 675:19 676:7,22 677:7 677:11,15,24 678:5,8 679:2,9,10,13,17 680:1,3 682:11,23 683:6,10,24 684:4,5,8,8,20,25 685:1,1 691:13 model-estimated 219:1 moderate 682:18 moderately 157:22 modern 15:25 141:11 279:9 modes 216:12,17,21 218:17 631:5 modification 9:12 88:13 107:6 142:15 171:20

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333:7,7,12,15,15 337:4,5 341:4,5 357:15,17 358:7 696:2 morning 2:1 3:6,9,23 4:16,19 5:6,17 6:2 8:2 11:25 13:11 14:12,26 17:26 33:9 34:5 35:20 55:15 71:12 79:12 204:7 247:17,17 248:8,9 249:5,17,19 249:25 250:12,21,24 251:7,12,20 252:25 261:24 277:26 297:3 302:9 303:14,25 304:21 316:13,16

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62:23 63:3 65:3 72:8 73:4,7,15,17,26 75:2 76:1 76:14,18 78:4 80:21,24 81:3,12,13 82:1 92:1 156:10,14,18 158:9,10,12 159:1,8 161:2 167:13 173:21 206:17 207:5,9,20,24 208:9 215:19 216:6 216:20,21,23 217:26 218:6,14,17 248:15,17,22 252:24 267:6 273:24 289:2,12 307:15 346:5,8 413:16,24 414:26 415:19 436:18 441:1 464:26 495:11 506:10 518:21 520:3 521:16,16,19,21 607:11 612:11 662:13,20 orange 302:15,16 orchard 305:24 361:20 363:23 orchards 109:1,23 111:20 302:12 305:20 307:22 order 89:18 90:7 93:15 95:19 96:17 100:2 101:22 108:3 109:18,26 111:22 112:17 114:15 121:21 122:15 149:25 154:26 164:20 168:15 170:20 171:2,17 172:22 183:7 184:9 185:13 191:8 194:6 216:17 224:1 227:15 228:3 233:13 258:13 260:4 263:7 269:7 277:20 279:17 284:17 331:2 367:5 396:7 398:12 401:20 425:19 426:3 449:6 450:6 450:13 561:1 635:21,24 650:2 664:1 orders 90:18 96:24 100:5 118:10 123:18 259:9

260:26 261:2 293:5 299:14 402:23 ordinary 283:3 organic 124:1 297:10 322:26 398:4,25 407:6 471:4 organism 24:7 163:14 167:12 199:16 314:24 374:8

443:2 449:26 459:26 462:22 473:25 497:4 504:13 514:23 527:5 533:9 625:13 642:25 organisms 4:6 24:5 165:10,16 168:23 199:20,23

206:22,26 207:3 213:2 245:24 266:14 301:23 423:19 462:26 497:5 510:4 514:5 532:20 558:12 558:14 587:4 609:13 611:13 620:24 624:25 635:1 675:23 organization 11:5 22:25,26 51:2,17 54:12 107:4 229:4,10 268:25 270:6,7 280:25 289:16 292:15 297:6 312:2,10 324:3 442:12 506:5 606:9 624:17 667:26 668:7 669:5 organizations 8:20 9:26 213:13 246:9 268:24

312:24 313:4 331:17 organized 36:8 organophosphate472:8 organophosphates 373:22 389:24 orientation 37:22 74:10 origin 160:2 186:13,18 original 94:19 397:25 originally 232:8 258:7 474:5 578:3 664:5 originate 121:24 or/and 632:9 Osmia 361:19 362:15 364:11 365:6 562:21 570:13

570:17 572:23 Ostiguy 5:21,21 250:16,16 336:23,24 339:19,20

344:2,3 345:18,19 348:21,23,23 350:23 352:21 353:23,24,25 354:19,19 355:19,20,20 356:4,12 356:13,13 357:7,7 364:22,23,23 371:13,14,14 376:22,23 380:11,12 385:9,10 387:14,15 388:26 389:1 445:3,10,18,19 451:22,24 457:23,24 461:7 461:8 462:14,15 467:26 468:1 471:8,10 472:13 472:14 473:10,11 475:22,23 478:3,4 479:24,25 481:25,26 483:21,22 485:6,7 488:1,2 490:26 492:13,14 497:13,14 500:25,26 502:18,19 503:7 503:8,15 504:24,25 510:11,12 516:19,20 532:22 532:23 537:1,2 540:12,13 550:7,8 555:18,19 564:22,23 565:9,10 570:23,25 577:2,19,20 587:18,19 590:10,11 592:15,16 594:26 595:1 600:1,3 617:18,19 629:10,11 635:8 636:11 660:24 689:22 ought 353:2 359:20 396:2,9 432:16 450:12 532:1

572:19,21 617:9 626:21 outbreak 322:19 outcome 51:1 61:13 147:10,19 157:20 229:3

230:17 231:1 290:18 436:20 439:17 440:6,12 447:19 488:24 509:7 511:14 522:12 536:19 576:21 614:2 628:17 636:13 647:6,15,20 648:19 655:4 666:2,15,17,26 668:14 673:1 674:4 677:5 679:19 680:6 687:19 outcomes 147:18 240:18 443:19 651:3,15 666:15

667:12 679:6 outdoor 57:19 outdoors 144:25 169:9 183:18 outliers 261:18 outlined 154:13 193:5 536:5 659:11 outlines 142:7 outlining 689:14

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276:23 oversight 325:17 overspray 352:1,9 438:11 overview 13:19 18:9,15,17 32:8 33:19 89:11 90:10 154:15 156:12 overweighs 545:21 overwhelm 582:21 overwhelming 168:16 629:13 overwinter 177:24 634:11 overwintering 62:1 139:18,19 173:10,17 176:21 176:24 185:10 189:23,26 192:5 227:4 589:7,8 634:8,22 657:2 overworked 659:7 over-predict 138:13 owed 284:9 owned 636:24 owner288:21 o'clock 622:2 P P550:19 package 220:4 519:25 637:5 662:12,16 packet289:17 packets 292:5,6 paddies425:3 521:7 524:14 530:2 paddy425:2 page47:22 269:13 291:9 295:16 296:1 304:17 305:2,12,14 306:9 334:17 407:21 553:13 554:14 paint-marked 634:17 pan 572:11 601:22 panel 2:2,9,13,17,23 3:23 4:7 6:19 7:11,13,17,22 13:12 14:4,6,24 16:8,9,24 17:21 18:4 23:21 31:21 31:25 33:6 71:11 84:24 87:22 89:1 150:8 194:23 203:9 215:5 221:21 244:10 247:26 249:10 251:16 251:22,23,26,26 252:8 263:16 274:24 279:15 280:5 281:18 288:18 296:12 299:24 301:13,24 302:7 313:1 318:24 325:5,10 334:9 336:26 340:26 342:25 344:5 350:9 352:23 353:16 357:2 364:21 365:16 372:6 377:6,17 380:13 382:5 385:11 396:13 403:22 405:15 407:9,24 409:2,6 415:10 417:5 422:25 425:24 435:7,16,19 436:8 436:14 444:3 446:7 455:3,15 459:2 466:24 468:4 469:5 475:25 478:5 482:1 483:24 485:17 492:18 493:12 503:25 507:1 512:8,14 513:24 533:10 538:6 539:15 543:9,23 551:8 561:16 565:8 571:14 573:14 582:5,12 584:19 585:15,19 587:24 596:4 598:20,24 599:1 602:6 607:1 608:9 614:26 616:17,23 617:3 620:8,18 626:2 627:2 631:26 643:8 658:13 667:11,14 669:12 671:6 675:14 681:17 683:16 685:10 686:7 687:4,18 689:13 690:23 691:3,8 693:2,19 694:19 695:10,11,13 Panelists 290:19 517:19 panels489:5 515:15 692:20 Panel's 302:5 422:10 435:5 511:16 582:12 paper7:1 8:8,14 10:17,19,21 11:4,10,12 12:4,11 13:1 14:8,18,20 16:26 30:13 33:10,14,17 34:4 41:26 48:8 63:6 64:16 65:15 66:4 70:11 71:17 78:14,16 79:5,26 80:14 81:18 82:19 89:8 93:1 99:10 101:11 108:2 121:14 126:1,4 128:25 140:15 152:4,26 154:13 155:3 156:17 201:20 202:21 205:6 206:7 208:3,19 211:20 212:13 213:19 214:16 215:12 221:10,12 224:9,11 230:1 231:21 240:9 255:4,16 256:14,20 257:2,15,18 258:21 265:22 267:12 269:13 270:14 272:9 278:11 279:15 285:26 288:4,14 289:14 293:17,25 327:21,24 332:4,5 333:20 334:3 335:22 336:4 338:26 340:3 357:20 358:15 376:12 377:11 395:6 395:10 397:3 398:13 399:21 402:14 403:20 410:15 411:12 422:22 425:15 438:20 442:3 443:1 443:23 450:18 465:14 476:16 482:9 490:20 491:9 500:3 503:21 508:23 513:25 515:26 522:22 523:6 536:5 538:24 540:8 543:21 545:16 547:24 548:23 553:13 562:9,23 567:12,19 588:26 593:14 618:20 619:10 620:5 621:3 622:17 623:5,19,24 636:22 644:12 647:24 650:21 651:14 668:14 670:22 671:5 672:18 674:11,21 678:6,8 686:3,5 687:1,14 689:4,20 690:20 691:3,22 692:10 693:22 694:7

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691:22 paradigm 641:4 parallel 659:12,21 parameter 667:18 671:9 674:6,7,8,15 681:25 parameterization 670:26 parameterize 679:11 parameterized 227:25 parameters 97:13,17,19 224:2 225:6 226:24

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28:13 44:18 48:15 57:26 60:11 67:7 69:20 71:17 79:23 128:18 130:13 132:12 133:6 136:8,10 140:14,17,17 142:10,22 144:21 152:17 153:9 156:6 165:12,18 167:20 168:2 188:21 197:5 201:26 209:11 223:17 229:22 233:18 234:8,12 240:15 243:26 246:13 276:12 283:17,18 290:4,19 314:10 320:4 333:4 337:11 341:11 350:4 351:26 359:4 363:9 366:16,22 373:5 378:23 383:9 384:7 392:12 397:9 402:7 410:13 413:4,13 414:17 417:21 418:5 423:12,17 424:10 425:22 439:19 446:23 451:5 456:9 458:10 484:2 515:24 526:17 531:3,21 538:20 541:24 557:4 577:24 580:7 590:16 598:24 619:25 627:9,11 629:4,12,20 639:12 640:12 657:3 664:12 681:6 682:6 687:18 689:13 692:9,15,20 693:20 694:3 partial 215:21 225:12 521:11 partially 221:24 222:1 516:22 675:4 participant2:25 participants 21:22,24 228:22 participate 16:8,24 355:12 participated 256:17 participating 267:11 271:19 274:14 696:13 participation 7:24 14:17 particles 41:12,20 299:22 349:15 434:2 particular 2:3 3:23 4:7 12:4 26:16 36:10 43:21 49:20 50:13 91:7,20 109:8 114:1 134:14 136:2 148:16 150:22,26 151:9 152:26 153:21 165:1 207:18 220:16 222:16,26 232:10 253:4 276:11 278:8 279:3,12,22 282:22,23 283:1 284:3 297:23 321:21 325:8,9 333:25 334:10 338:8 342:8 345:6 345:18 346:20 355:22 359:23 360:14 363:2 366:23 367:18 368:15 370:3 381:23 399:22 401:13 402:13,23 410:21 411:11 421:15 442:17 444:11,24 445:22 449:4 451:15 461:20 472:10 486:16 493:23 512:3 526:8 540:6,10 544:23 548:21 549:11 553:16 561:17 601:25 605:18 608:8 610:10 615:23 622:18 623:2 624:15 629:16 630:3,19 636:13 642:8,14 666:5 667:17 669:7 670:23 672:11 678:15 679:7 689:15,20 694:8,9 particularly41:21 137:7 139:17 151:9 193:3 199:13 217:19 238:3 260:15 272:9 273:5 284:15 295:11 317:6 358:13 364:2 387:26 398:23 401:7 411:14 431:3 437:24 442:13 467:18 475:1,5 516:6 517:25 606:4 613:13 625:17 657:7 668:9 680:23 681:13 particulars 359:21 644:2 parties 419:11 partitioning 120:21 partitions 347:21 partly 338:11 552:22 partners 10:15 21:13 25:19 31:24 parts 113:11 117:12,14,15 247:12 256:13 258:12

298:12 332:5,7 402:20 404:4 411:6,13 423:22 424:3 425:11 596:25 600:10 643:24 656:13 pass 14:23 103:25 142:18 214:24 217:6,9,14 218:3

272:22 273:1,10 282:14 287:21 427:17 610:19 649:17,17 passed 294:7 passes 165:9 166:16 passing 242:3 436:20,21 passive 125:16 patently 666:19 path 50:20 378:15 619:2 pathogens 299:2 471:11 505:3 pathologist5:25 pathway 38:21 51:1 58:24 63:10,14 77:4 80:8,17 135:4 173:22 180:13 396:10 411:2 424:21 425:21 511:15,18 512:3 648:9 657:8 pathways 37:12 41:9 47:24 55:4,6,7,11,17 71:16

79:3 81:21 171:22 229:3 408:7,11,11 436:15,17 465:21 509:8 511:14 513:22 648:8 668:14 672:12 677:26 patience 687:2 pattern 57:8 144:20 251:24 255:22 283:24 569:15

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317:1 586:2 patties 172:4 174:14 paucity 395:2 Paul 5:4,4 231:10,19 232:5 250:3,3 349:22 366:4,5

372:12,26 373:11 392:20,20 397:15 401:19,23 402:3,13 404:3 405:26 686:25 pause 68:12 333:8 pay 304:11 308:10 310:4 311:4 paying 310:5 peak 19:12 169:23 227:2 234:17 545:14 peaks 235:14 peer2:15 7:5 107:17 281:13 312:21 395:5,8 575:1 peer-reviewed 162:14 PEIP23:8,10,12 Pellston 10:22 22:3,8 29:5 65:25 156:21 202:23

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263:15 276:8 282:7 311:15 312:4 313:25 314:1,6 314:9 322:10,21 323:21 324:9,26 340:10 342:20 342:24 418:20 419:18,21 430:23,25 475:2 546:6 547:17 558:12,13 629:21 664:1 685:6 692:6 694:4,14 peptides 449:17 perceived 201:20 percent 19:26 20:3,6 69:22 79:24,24 96:15 102:5 102:15,16 109:17,24 135:5 157:8 158:3 178:15 186:13 187:25 191:19 208:2,5,7 217:7,13 234:23 260:8,9 262:3,7 263:1,3 286:5 297:15,16 306:24 307:2 317:14 320:15 321:11,16,22 332:12 337:20 337:20 375:8 377:12 389:20,21 395:12,26 425:19 425:19 438:13,14 471:1,1 477:9 482:15 494:15 499:18 500:10,17 501:5,7,13,17 502:13,15,20,24 503:1,9 548:7 552:13 554:3 599:11,19,20,23,24 600:23 601:13 608:18 610:7 612:4,6,6,18 percentage 79:20,22 186:2 320:26 321:13 600:1

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261:7,9,12 398:13 399:16 494:25,25,25 perception 287:17 Percival 696:17 Perdue 6:3 perfect514:18 534:25 605:11 perfection 12:22 perfectly 522:13 592:26 perform 312:11 472:9 667:8 performance 173:4,17 177:18 178:10 298:4 performed 176:3 226:15 performing 151:14 period 3:5,9 92:11 101:16,21 131:23 132:14 135:23 159:21 169:25 170:4 176:16,23 183:26 190:17,18 192:2,3,5 222:12 238:7,10 247:18 248:14 252:7 289:6 297:15 317:2,17 352:8 370:19 379:1 444:2 453:15 468:11 477:8,10 479:10 482:14 483:2 485:20,22 488:25 523:16 530:15,20,20 532:14 534:15 537:9,12 543:25 544:18,19,23,24 545:2,4,6,12 569:13 586:14,15 588:18 589:7,8 590:8 594:19 637:7 662:4 685:9 periods 52:9 173:24 176:21 428:2 488:23 523:14

589:19 permanent 87:21 permanently 197:25 permitted 208:9 216:4,8 persist506:18 persistence 199:13 634:24 656:25 persistent 184:2 188:25 192:4 199:17 person410:3 469:15 545:19 557:9 651:15 660:15

664:16 688:20,25 personal 263:21 363:9 557:18 573:25 574:24

588:24 688:17 690:3 personally46:21 47:20 316:5 518:8 523:1 557:17

574:21 696:14 perspective 295:3,6 312:4 406:18 428:12 502:16

611:6 614:1 pertain 377:21 pest 13:24 14:13 15:2,25 32:15 191:23 200:17

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453:8 479:14 499:1 525:1 627:25 629:8 639:22 639:25,25 practicality 498:24 577:15 practically 157:21 practice 116:4 133:11 188:21 193:11 313:2,15

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25:17,17,18,23,25 26:6,9,13,20 27:4,17 31:14 32:10 33:7 38:12 45:14 48:16 50:2,5,23 51:3,14 51:24 52:11 53:18,26 54:5,6 55:17 56:11,16 57:6 58:3 59:7 60:8,15 61:19 63:10 64:20 65:11,15 66:14,16 67:16 76:6 81:17,22 84:6,10 130:6,25 136:5,8 137:2 139:9,10,11,14 140:22 143:15 144:21 152:9 153:9,22 154:17 155:2,21,25 163:23 165:12,18 175:7 185:3 197:5 199:17,19 200:23 201:26 202:2,26 203:13,25 204:4,8,18,23 205:16,26 208:20 209:16 211:11 212:22 214:14 214:19,24 219:11 240:19,20 253:12 254:21 255:4 267:11 274:14 278:24 279:10,15 280:1,2,12 282:5 289:15,15,21,22 290:4,19 294:18 300:17 318:25 323:23 327:1 386:11 391:7,8,13 395:23 396:4 399:2 406:26 414:12 436:11 438:19 453:2 509:11 534:13 536:22 537:17 545:7 567:11 607:25 618:25,26 622:26 629:6 643:18 647:19,21 653:19 654:8,20 657:21 659:11,15 660:22 661:6 663:10 664:26 665:18 681:14 686:19 687:21 696:9,13 processed 31:22 38:25 45:4 47:1 74:14,15 75:13

93:4 361:8 378:24 388:16,20,21,22 391:23 467:11 487:20 488:15 537:11 processes 23:23 39:10 46:18,22 77:13 130:1,2 135:10 221:23 222:8 225:6 226:5,10 227:17 254:5,22 272:5 279:6,13 280:13,17 648:3 processing 38:14,16 46:13 74:1 75:12 145:25

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434:14 511:9 recommendation499:9 500:9 580:19 585:7 recommendations 2:18 24:1 133:2 192:6 208:10

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276:1,18 334:16 349:12 395:11 410:10 421:9 423:23 425:26 437:24 438:1 462:2 562:15,25 582:3 611:22 674:6 regarded 292:14 457:3 520:14 regarding 22:5 26:17 31:25 34:20 35:23 43:5

55:26 57:7 135:1 137:12 145:16 150:10 155:3 164:26 179:17 182:7 200:8 201:21 213:16 312:16 313:5 316:22 319:16 415:4 425:2 458:3 465:4 478:9 497:2 538:16 652:9 667:24 670:20 regardless 35:12 463:24 553:3 579:8 645:24

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164:9 222:10 240:21 241:9 318:22 511:5 647:3 666:9 668:15 671:11,13 675:7 relative40:26 46:5 49:25 55:9 71:14 78:20 79:8

98:19 100:21 102:17 104:10,11 105:25 124:13 128:10 172:15 173:25 178:7 181:14,20 182:23 187:17,23 190:17 191:6 192:19 193:15 200:9,11 214:13 235:13 254:11 266:16 369:19 383:23,25 383:26 384:22 403:17 411:7 412:10,22 424:4 479:2 499:21 504:9 513:8 540:26 555:17 563:3 589:15 614:1 relatively 50:6 137:11 143:20 165:7 167:3 168:25

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193:11 222:24 551:4,6 569:5 574:4 579:23,25,26 581:10 583:7 584:3 630:24 631:21,21 633:25 635:19,21 639:17 replications 518:19 report2:25 7:4 9:7 18:26 19:4,7 86:10 161:4

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163:8 178:11 211:1 234:23 292:20 297:13 298:26 325:24 326:6 346:26 365:26 366:26 395:11 402:22 463:23 497:10 621:11 623:23 reporting 260:16 292:13 293:10 311:9,11 325:15

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225:16 323:23 582:22 representations 72:16 94:12 388:14 representative 92:11 99:18 101:9 105:4 111:11

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93:10 97:15,20 99:5 101:23 139:6 140:18 142:22 156:7,12 167:21 168:1 190:12,19 239:4 255:15 267:16 279:13 286:24 312:9 328:25 369:19 428:9 492:25 596:12 661:22 663:26 requires 83:17 156:9 165:21 284:16 482:19 515:9 requiring 217:7 274:2 reregistration 323:15 research4:5,21,25 5:7,8,15,19,19,22,26 6:1,3,6

21:6 23:5,18 77:7 88:1 164:14 211:24 246:8 247:11 249:24 250:6,11,13,14,19,20 259:3 274:13,17,21 283:16 300:15 311:24,26 312:1,1,6 312:9,10,10,18,19,24 313:4 322:3 324:5 340:11 343:20 346:26 352:5 358:17 410:3 413:7 443:21 460:14 492:25 509:18 555:8 623:12 668:1 researched410:17 researcher420:26 683:11 researchers 254:6 299:3 451:1 533:17 researches 215:17 reserve 290:18 563:15 reserved 66:16,20 67:9 reserves 20:10 39:16 187:9,11 360:20 534:5,6 reservoir 653:22 resident 323:14 residual 167:23 181:5 residually430:1 residue 59:26 60:1,22 110:22 113:6 114:1,19 119:8

141:23 143:5 145:15 146:18,25 147:19 150:2 157:25 164:20 174:10,15,19,21 179:9,10 192:9 192:10 194:19 206:5 261:7,16 329:18 339:11,12 339:13 349:24 373:6 378:14 381:18 386:3 392:14 395:2 396:19 397:2 428:22 439:2,3 453:5 535:1 539:3 571:4 594:21 residues 23:22 24:25 25:6 37:6,7,11,18,26 38:10

39:25 40:12,26 41:5 46:26 60:22 72:25 73:1 86:3 86:14 98:17 109:4,14,18,22,25 110:19,24,26 111:2,5,11,16 112:15 113:14 117:12 130:24 133:9 137:12 143:13,23 144:18 145:20,23,26 146:20,22,23,26 147:2,23,26 155:15 157:26 158:6 170:19 172:12 174:4,8 181:11 186:4,7,10 186:16 191:15,17 192:4 201:11 202:13 206:6 277:4 298:11 299:9,16,21,26 300:4,7,24 301:5 335:11 338:9 348:2,4,6,15,16 349:1,2,5 351:1,11 352:19 366:7 373:26 390:10 393:5 394:14 402:15 404:13 405:22 412:5 431:21 439:25 535:16 538:17 656:24 657:14,16 resilience 188:15 resins 73:2 338:13 350:13 resistance473:4 664:6 resolve 661:3 resolved 12:10 360:25 resolving 9:25 85:5 resource 314:10 resources 26:17 31:3 50:7 146:11 175:4 181:19

192:23,25 242:4 244:7 255:13 271:21 283:16,20 352:8 475:12 498:22 523:17,17 541:3 668:11 resource-intensive 181:14 respect28:12 31:2 33:18 203:24 212:7,25 213:4

256:14 258:6 275:9 292:17 482:20 517:26 546:21 respective 91:26 respectively 128:3 129:15 217:7 respiration 506:25 507:8,13 respirator 653:23 respiratory41:13 653:25 respond 32:20 139:4 379:13 505:1 responding 344:7 670:3 response 21:9 32:24 47:25 48:12,22 68:5 69:26

71:5 87:12 140:3 173:15 174:3 194:8 198:2 199:4 199:4 202:8 203:5 207:2,8,9,15,19,22,23,25 208:2 243:24 245:11 292:15 378:6 379:16 382:6 389:13 435:5 439:20 449:17 450:21,24 454:13 455:22 458:14,15 473:3 475:21 491:23 492:22 496:14 504:3 505:16,21,22 506:4 511:16 520:16 520:23 550:24,25 584:1 588:12 595:7 604:9 619:18 656:12,14 675:8 677:8 680:17 691:4 responses 35:16 39:11 48:9 67:4 386:12 465:22

470:9 482:17 499:24 505:2 509:14 677:25 responsibilities 313:7 629:23 responsibility 238:19 245:9 597:16

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90:1 95:5 97:21 110:1,3 122:7 139:13 152:10 164:26 165:17 170:6 174:1,23 193:7 217:25 218:7,23 223:2,5,5,8,8,26 224:3 227:1 228:2,19 232:2 254:7 268:20 270:23 271:7 275:16,17 305:12 306:12 308:22 310:2 343:7 383:9,10 469:22 486:15 509:24 529:13 533:6 548:25 563:13 569:26 575:25 576:14 588:15,20 589:2,18 591:7,20,25 595:24 633:23 638:21 663:23 684:26 resume 87:15 retribution 326:9 return 112:24 141:8 186:3,21 298:7 435:19 462:9

479:2 553:24 560:3,5 returning 146:1 462:6,11,17 536:22,24 537:17

569:11,12 601:15 returns 49:26 Reuben 140:5,6,7 206:1 209:18 214:12 215:8,10

215:11 219:20 696:3 reveal 174:14 reverse 207:17 401:20 review2:16 17:4 75:24 77:12 97:2 111:13 113:14

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274:11 312:21 404:8 reviewing 155:19 318:25 406:1 552:6 676:21 687:18 reviews 7:5 279:4,5 revise 661:21 revised 293:10 357:4 rewarding 17:23 rewritten 335:26 re-apply 654:26 re-asked 456:8 re-looked 307:17 re-queening 307:2 ribosomal475:15 rice425:2,3 rich451:3,17 467:4 Richard 14:14 16:18,19,20 696:7 richness 333:19 335:22,23 337:8 338:19,24 339:4

648:10 rid 637:18 639:14,15 ride 360:2 ridiculous 574:9 638:6 ridiculously 639:7 right 3:11 34:6 43:7 46:12,17 48:23 51:12,17 60:16

68:11 71:7 113:26 126:18 131:18 132:6 133:17 133:21 138:20 165:13 219:22 241:13 247:24 252:5 256:11 277:12 295:9 296:13 307:5 308:19 309:11 310:4,9,11 312:5 315:20 320:8 321:23,26 327:26 332:19 334:6 336:25 337:3,3,16 339:6 341:2,3,17 353:26 354:22 357:13 365:19 373:2 378:17,20 382:14,16,26 383:1,26 387:2 388:9 391:16 392:8 427:14 430:11,12 434:22 437:14 441:7 460:2 471:21 474:12 476:4,6,16,25 480:2 480:19 493:17 494:16 501:17 502:9 503:18 508:14 510:21 512:5,19,22 522:6,14,24,26 523:24 532:10 536:17 542:18 545:4 562:10 565:26 566:9,15 572:10 582:1 583:3 587:25 599:22 601:16,19 609:26 611:23,25 612:22 613:2 618:19 621:17 627:7 630:11 634:20 641:18 645:8 646:11,20 651:26 652:4,5 661:26 667:4,19 671:19 673:11,12 676:16 684:12 693:5,10 rigid 56:21 rigor 253:13 rigorous 280:2 292:2 ring 159:25 316:6 380:26 ripe 184:14 risk 2:4 3:24 5:13 6:23 7:16,16,19 10:6,13,25 11:9

11:11,13,14,17,26 12:1,3,5,6,10,15,18,23 13:17 15:14 18:5,11,17,22 21:14 22:3,6,14,17 23:15,16 23:22 24:7 25:16,21,22,22 26:1,2,3,9,10,10,12,14 26:14,20,23,26 27:1,2,2,9,19,20 28:2,4,23 29:6 29:12,24 30:6,22,25 31:3,14,23,25 32:17 33:8,12 33:20,22 34:10,11,20,25,26 35:1,18,21,23 36:1,3

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193:19 459:23 556:15 566:12 586:3 587:9 626:14 626:18 robustness 446:26 672:25 673:1,5 rocket 308:8 Rod 316:11,13,13 319:10 320:3,18 321:2,6,18,23

321:26 Rodenticide 28:16 role 9:18 10:5,8 18:13 21:11 23:25 236:9 238:12

238:21 279:14 280:1 289:23 451:20 551:20 554:13 596:8 651:20 669:2 rolling 684:7,9 room 3:26 138:16 198:1 238:2 248:1 257:20

296:23 375:13 410:22 681:10 rooms 248:3 root253:23 317:6 632:6 roots 120:6,16 149:14 291:6 350:6 Rortais 96:8,13 97:3,5,14 101:24 262:19 Rosalind 5:24,24 32:13 45:24,24 46:7,13 81:9 131:17,20 150:12 151:1 235:4 250:18,18 324:2,6 325:1 329:7,7 333:26,26 337:16,17 340:1,1 341:17 350:18 354:11 355:1,15,15 356:21 357:25 357:25 358:3,20 365:8 368:16 370:5 375:25,25 381:4 382:14,20 385:13 387:7 389:15,15 419:25 433:3,6 434:7,20,24 440:16 441:6,6 455:21 457:9 468:5 469:8,8 474:2 476:1,23 480:11 481:18,18 482:25 484:21 485:19 486:23,25 489:16 490:13 490:23 492:5 495:19 500:22 504:18 507:5,17 510:7 527:20 529:17 535:12 537:16 559:5,15,23 565:22 566:14 571:16 574:8 578:8,8 580:13,24 581:15 583:17,17 584:8,13 585:3 596:6 598:6 618:14 619:6,6,11 633:15 641:14,20,26 651:20 652:3,6,22 654:16 669:22 674:18 679:23 683:5 690:14 Rose469:15 636:23 rotundata 363:9 rough 218:24 376:9 584:10 roughly 19:6,13,23 20:2 147:13 234:17 395:26

612:3

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443:20 row 104:25 225:9,15,19,25 226:5,8 322:21 rows 601:25 royal 39:1 44:23 74:2,11,16 75:13 76:17 93:5 95:5

95:6 99:23 101:18 102:26 103:24 104:7,12 105:14 137:21 138:1,9,16 159:19 160:8 338:8,10 355:9 356:7,8,9,17 378:24 379:11,20,23,26 380:3 380:6 381:13 382:21,25 383:13,22,25 388:5,15 388:21 389:5 391:24 433:16 465:3,6,14,17 466:4 466:4,12 469:5,5,6,11,15,16,16,20,20 470:1,9,12 470:17,20 471:2,3,6,11,15 RQ 58:9,14 59:8 60:4 63:23 67:17 201:2,4,5

206:14,18 208:13 209:8,14,19 210:10,25 217:16 217:19 218:14 219:25 256:24 491:12 492:10 RQs 206:2 346:5 RT25 158:3,4 rubber 322:8 rule 277:19 420:14,15 584:18 ruled 59:24 382:25 rules 574:10 643:1 run 161:9 216:3 304:12 310:3,4 324:8 426:16

428:19 453:18 456:11,16 462:3 480:1 566:22 693:14 running 198:1 215:4 230:16 288:22 305:10 306:24

310:5 506:2 541:2 684:2 691:9 runny 653:26 runoff40:24 82:5 318:4 321:9 426:14 runoff/erosion 37:9 runs 307:12 rural 306:13 rush 139:20 Rutgers 6:6 250:25 Ryan 120:20 126:3 398:1,22 Ryan/Burghardt406:11S s 87:19,19,19 127:26 Sacramento 684:23 sacrifice453:24 sacrificing453:21 sacs 147:20 safe 10:10 256:3 262:12 273:25 288:3 323:5,6,8,13

324:22,25 529:10 668:26 safeguarding 254:16 safely 693:18 safer 15:25 279:8 safety 11:2 23:20 29:13 64:11 156:22 213:15

287:20 288:25 289:2,16 292:16 324:23 436:26 663:20 sake 605:6 salamander 664:3 salamanders 664:2 salesmanship 663:17 salient 653:6 sample 149:2,5 182:19 261:20 301:2 305:6 313:16

313:17,20 453:22 497:15,16,17 498:12,20,22 515:2,9 536:21 537:17 538:3 540:16 541:17 548:26 549:1,3,6,21,26 572:10 580:25 625:24 630:22 636:25 639:7,17 sampled 293:20 581:19 samplers 434:1 samples 149:12 303:23 304:12 305:1,6 310:3,3

311:15,17,20 327:12 396:21 412:6 537:3 635:21 sampling 148:25,26 149:14 186:16 191:16 224:13

332:13 439:18 537:25 572:11,12 580:26 Sandy4:11,11 136:21,22 390:2,3 635:3 661:4

663:10 685:12,13 686:1,4 SAP2:14 3:10,21 7:11,13 48:16 70:17 81:2 105:22

106:2 128:22 129:1,4,10 140:10 197:4 199:16 208:25 209:2 229:12,16 249:15 251:8,10 283:23 285:13 290:19,19 291:11 292:2,9 294:7 641:3 693:17 696:15 Sappington 32:5,25 33:1,2 42:20,26 43:7,17 44:4

44:17 45:1,7,18,20 46:3,12,17 47:16 48:7 53:3,4 66:26 69:15 71:24 84:3 89:15 91:6 143:5 152:12 199:9 200:25 204:7 208:18 214:18 219:9 220:24 221:1 230:7,23,23 231:17,20 232:9,9 240:1,8,8 244:4,4 330:10,10 345:12,12 353:18,18 354:2,2 354:23 355:5 357:1 422:12,13 432:7,7 436:5 437:12,12 441:12,14 497:20,21,21 527:11,12,12 530:6,7 538:14,15,15 552:2,2,21,21 553:19,19 561:26 624:8 626:4,7 630:3,13 643:9,11,16 669:26 670:19 676:12,14,19 683:18 696:3 Sappington's 48:12 482:4

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www.transcriptionetc.com Page 109 satisfactory 477:21 satisfiable 240:25 satisfied 2:22 54:14 satisfy 187:26 save 308:24 309:2 333:1 335:19 430:23 saw43:10 69:23 129:25 302:23 303:2 324:14

437:16 499:5 619:24 661:6 saying 16:13 48:12 90:8 132:10 136:15 239:13

277:14 321:16 327:22 329:11,13 335:18 344:16 352:17 389:26 424:18 427:4 461:21 466:18 482:18 495:26 519:18 557:9 559:11 601:22 607:18 609:10 620:9 635:4 637:25,26 645:8 647:24 650:10 652:9 653:14 679:12 681:9 685:2 says 283:13 323:5 334:24 335:2 340:4 402:15

414:23 439:2 554:1 648:14 652:2 659:13,18 673:16 scale 27:25 149:17 222:23 421:6 462:21 511:5,11

588:6 598:9 606:6 623:24 649:26 650:11,11 673:7 677:19,20 scaled 116:2 scales 190:24 scaling 372:10 619:16 scarce 177:5 scary470:22 scenario 20:11 83:2 190:15 245:11 246:25,25

268:2 295:3,5 379:18 382:2 388:20,25 404:25 424:17 437:10 438:10,12 487:24 568:7 677:4,5 scenarios 121:3 130:9 156:5 213:3 246:13 265:17

268:19,19 295:9 403:24 404:5 430:16 437:10,13 488:18 568:4 590:1 scene 85:25 schedule 87:13 407:13 scheduled 87:14 scheduling 26:17 schematic44:2 scheme 64:12 65:4 83:7 148:25 223:7 659:1 schemes 64:9,18,25 65:4,9,24,26 66:9 262:8 Schertz 285:10,12,15,16 288:18,19 Schlenk 3:20,22 4:3 6:14,18 13:6 14:24 16:17

17:25 32:7,19 42:14 44:11 45:22 47:7 48:20,26 52:26 66:18 67:11,26 68:15 69:12,23 70:3,9 71:3 81:7 82:3 83:3 84:12 85:6 87:11,21 88:23 89:1 129:21 130:19 131:16 132:15 134:25 136:21 137:16 138:21 140:1 150:9 151:21 152:11,23 154:3,7 194:24 195:10 196:6 197:15 199:7 200:1 200:5,20 201:18 203:3 214:20 215:3 219:12 220:1,21 229:24 231:9 232:15,20 236:20 237:16 239:14 242:20 243:11 245:5,14 247:15 249:9,13 251:4,18,19 252:5,6 274:23 275:21,25 276:16 277:22 280:4,18,20 284:6,21 285:8 288:17,20,24 296:11 302:6 309:4,20 311:3,7,22 314:16 315:8 315:22 316:8,11 319:2,13 320:23 321:14,24 322:1 323:25 325:2 326:21 328:8 329:6 330:9,22 331:6,26 332:14,17,20,26 333:10,14,24 336:16 336:19,21,23,25 337:14 339:7,14,17,19,26 340:14,25 341:15 343:2,10,22 344:2,5,12,18 345:2,17 348:9,12,18,21 349:7,20 350:9,16 352:14,21 353:15,22 354:10,25 355:19,25 356:4 356:12,20,25 357:10,23 358:2,5,19 364:4,7,17,21 365:2,7,15 366:2 367:24 368:14 369:11,21 370:1 371:3,13,18 372:5,16,24 373:2,10 374:4 375:16 375:24 376:22 377:5,16 378:16 379:7 380:11,13 381:3,7,21 382:4 383:15,19 384:2,12 385:9,11,21 385:26 386:7,16 387:6,14,23 388:7,17,26 389:6 389:12 390:2,7,15,20 391:14,19 392:2,8,18 394:10,19 396:13,23 397:5,14 400:12,25 401:15 401:18,19,22 402:1,4,12 403:7,9,11,14 404:2 405:9,12,14,17,25 406:22 407:1,9,12 408:2,8,14 408:18,22 409:2,11,16 411:9 412:8,26 414:21 415:8,15,20 416:9,26 417:8,14 419:4,23 420:3,22 421:13 422:2,8,16 423:5 426:19 427:2,5,8,20 428:15 429:16 430:6,19 431:7,23 432:1,10,17 433:1,12,22 434:5,18 435:2,25 436:5 437:4 438:21 439:6,10 440:4,14,22 441:11,16,24 445:8 445:16 446:1,7,19 451:7,22,26 452:14 454:2,15 455:1,20 456:21 457:8,16,23 458:2,7,23 459:13 460:3,9,21 461:7,23 462:1,13 463:4 464:2,16,20 465:7 467:22,26 468:3,18 469:2,13,23 470:3,14 470:23 471:8,13,18,21 472:1,11,13,16,20 473:1,7 473:10,13,16,26 474:8,12,18 475:18,22,25 476:5 476:20,25 477:6,26 478:3,5,14,17 479:8,24 480:9 480:17 481:2,16,22,25 482:1,11,24 483:15,21,24 484:7,20 485:3,6,8,17,25 486:6,12,22 487:11 488:1,13,20 489:7,15 490:12,22,26 491:6,13 492:4,13,16,18 493:4,10,18 494:11,19 495:18 496:10,17 497:7,13,19 498:8,14,18 499:4,10,15 499:23 500:21,25 501:2,10,16 502:3,10,18,25 503:7,16 504:1,17,24 505:8,13 506:7,21 507:3,10 507:16 508:1,7,13 509:9 510:6,11,15,18,21,26 511:13,23 512:5,17 513:12 516:18 517:8,19 518:3 519:16 521:1,8,25 522:16 523:4,22 524:23 525:5,23 526:5,25 527:11,19 528:5 529:16 530:6 530:23 531:4 532:21,24 533:10,26 534:20,22 535:6,11,19 536:2 537:1,7,15 538:6,11,21 539:11 540:12,21 541:6,22 542:3,17 543:1,13 544:26 545:23 546:4,17 547:4,9,16 548:5 550:7,16 551:8 551:16,25 552:17 553:8 554:6,21 555:18,22 556:1,20 558:7 559:4,14,22 560:17 561:15,24 562:1,10 564:22,24 565:5,8,20 566:16,25 567:13 567:20 570:23,26 571:9,14 572:25 573:5 574:6

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560:8 696:4 scope 53:22 286:3 336:3 544:25 550:4 Scott285:10,12,15 288:19 scout 604:15 scouting 573:2 604:4 screen 13:26 26:21,23 54:1 77:18 118:8 119:1 123:10 129:12 139:7 142:19 153:26 165:9 166:17 171:18 175:15 199:22 215:14,15 216:2,3,25 217:3,5,6,10,14 218:3,8,8,16,22,24 255:12 258:26 261:4 262:11,12 272:15 384:8 403:25 441:23 461:21 484:6 490:9 608:4 610:25 611:21 616:23 screened 134:15 265:17,23 271:11 screening 50:6 52:1,11 54:6 59:24 63:1 65:7 67:23

68:21 69:1 83:5 107:3 113:9 117:7,9,26 118:6,9 118:18 119:7,23 123:3 126:5,9 127:24 136:13 137:6 150:15 153:5,13 155:14 156:1,13 159:16 164:26 165:2 166:6 167:1 169:1 175:14 178:16 194:1 196:10,12 198:12 201:14 205:16,24 206:9 208:24 211:21 218:9 272:12 385:2 402:16 413:23 424:13 443:10,12 447:21 456:2 475:17 619:3 624:24 626:21,24 668:9 screenings 443:7 461:6 screening-level 405:22 474:17 614:13 screens 103:26 403:20 514:17 screwed 324:18 se 20:14 135:18 233:26 284:23 353:14 429:18 sealed 342:22 556:14 searched 327:6 searches 327:6 season 160:2 317:1,7 318:10 338:4 344:15 502:13

588:14 631:21 seasonal 223:3 430:6 588:10 589:3 seasonality445:1 588:12,19 seasons 359:14 seats 296:18 512:23 second 36:4 53:2,5 56:15 58:11 64:12,24 77:25

83:10 84:14 86:12 98:2 134:8 143:2 147:11 162:20 172:18 177:9 182:11 183:10 186:5 188:17 209:4,7 222:14 225:15 227:20 243:26 244:14 251:8 296:19 298:24 305:8 310:15 338:5,18 342:14 383:9,15 384:4 410:13 413:4 414:17 423:12,17 424:10 425:22 446:10 471:22 479:22 479:23 519:23 553:21 secondary 38:3,5 245:2 secondly 79:16 152:17 323:14 396:8 406:5 531:22

570:20 secreted 349:4 secretions 93:7 388:23 section4:13 33:14,17 48:4,6 71:18,18 89:7 101:11 128:24 136:11 140:14,14 185:1 190:2 201:13 221:9 231:8 294:12 333:19 381:12 392:13 397:10 402:8,14 446:10,12 447:10 458:8 472:17 508:23 510:17 538:24 542:18 547:24 567:15,18 593:14 670:21 sector 688:10 security 287:25

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