Document ID: EPA-HQ-OPP-2012-0507-0004
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2014-05-28T04:00Z

BIOPESTICIDES REGISTRATION ACTION DOCUMENT
                                       
                                       
                                       
                                       
                                       
                                       
                                       

                                       
                                       
                        Clonostachys rosea strain 321U
               previously known as Gliocladium roseum strain 321U
                                       
                     Pesticide Chemical (PC) Code: 021000
                                       
                                       
                                       
                                       

   
                     U.S. Environmental Protection Agency
                         Office of Pesticide Programs
                Biopesticides and Pollution Prevention Division
                                       
                                       
                                 May 22, 2014
                               TABLE OF CONTENTS
I.  	EXECUTIVE SUMMARY	4
II.  	ACTIVE INGREDIENT OVERVIEW	5
III.  	REGULATORY BACKGROUND	6
	A.  	Applications for Pesticide Product Registration	6
	B.  	Food Tolerance Exemption	6.
IV.  	RISK ASSESSMENT	7
	A.  	Product Analysis Assessment (40 C.F.R. § 158.2120)	7
	B.  	Human Health Assessment (40 C.F.R. § 158.2140)	8
	C.  	Environmental Assessment (40 C.F.R. § 158.2150)	9
V.  	ENVIRONMENTAL JUSTICE	10
VI.  	RISK MANAGEMENT DECISION	10
VII.  	ACTIONS REQUIRED OF THE REGISTRANT	11
	A.  	Final Printed Labeling	11
	B.  	Terms of Registration	11
	C.  	Reporting of Adverse Effects and Hypersensitivity Incidents	12
VIII.  	GLOSSARY OF ACRONYMS AND ABBREVIATIONS	12
IX.  	BIBLIOGRAPHY	13
	A.  	Studies Submitted to Support the Clonostachys rosea strain 321U  Pesticide  Product Registrations	13
	B.  	Environmental Protection Agency Risk Assessment Memoranda	15
	C.  	Other References	15
APPENDIX A.  MICROBIAL PESTICIDES DATA REQUIREMENTS	17
APPENDIX B.  PESTICIDE PRODUCTS	29
                                       
                                       
                                       
                                       
                                       
                                       
                BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM
                                       
                                       
                      Office of Pesticide Programs (OPP)
                Biopesticides and Pollution Prevention Division
                          Microbial Pesticides Branch
                                       
                                       
                                Science Review
                                       
John L. Kough, Ph.D. 		Product Analysis, Human Health
Shannon Borges		Ecological Effects
		

                               Regulatory Review

Kimberly Nesci		               Chief, Microbial Pesticides Branch
Shanaz Bacchus		               Regulatory Action Leader
Shannon Borges 														   Microbial Pesticides Team Leader

											U.S. EPA-PMRA Coordinator
Gail Tomimatsu, Ph.D.												   USEPA PMRA Coordinator
I.  EXECUTIVE SUMMARY

Background
On March 25, 2012, W.F. Stoneman Company, LLC, on behalf of AEF Global, Inc., 201, Mgr-Bourget 
Levis Quebec, G6V 6Z3, Canada, submitted applications for two pesticide products to the U.S. Environmental Protection Agency (EPA or the Agency) under section 3 of the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA). These pesticide products, FPSolano[(R)] Technical (EPA File Symbol: 89046-E), and an end-use product (EP), FPSolano[(R)] (EPA File Symbol: 89046-R), contain a new active ingredient (a.i.), Clonostachys rosea strain 321U previously known as Gliocladium roseum strain 321U. The application was reviewed jointly with Canada's Pesticide Management Regulatory Agency (PMRA). Both PMRA and the EPA scientists reviewed product analysis, acute mammalian toxicology, and nontarget organism data and information (40 C.F.R. §§ 158.2120, 158.2140, and 158.2150, respectively) submitted to support the registration of the two proposed Clonostachys rosea strain 31U pesticide products. Summaries of these reviews are available in Appendix A in this Biopesticides Registration Action Document (BRAD). 

Clonostachys rosea strain 321U is a Microbial Pest Control Agent (MPCA) that is intended as an antisapstain product for control of blue stain and mold on freshly cut lumber and logs in hardwoods and softwoods. FPSolano[(R)] Technical, which is provided as a solution, is proposed for formulation into the wettable powder EP FPSolano[(R)]. When applied, the pesticide displaces wild molds and staining fungi from taking up their natural ecological niches on sawn wood products and logs, thus allowing the MPCA to become established before the pest species. The MPCA metabolizes simple, non-structural nutrients present in woody plant materials, thereby minimizing nutrients for staining fungi.

Clonostachys rosea strain 321U, a fungus in the family Bionectriaceae, was isolated in Ottawa. It colonizes living plants as an endophyte, digests material in soil as a saprophyte and is also known as a parasite of other fungi and of nematodes. Its identity was confirmed by Dr. K.A. Seifert at the Centre of Land and Biological Resources Research (Agriculture Canada). Clonostachy rosea has a cosmopolitan distribution.  It is common in a wide range of habitats in tropical, temperate, sub-arctic and desert regions of the world.  It has been reported from cultivated grassland and woodland, forest, heathland, freshwater and coastal soils.  This fungus has been frequently associated with cysts of Heterodera sp., Globodera sp. and other nematodes in soil and with sclerotia of Sclerotnia sclerotorium, Phymatotrichum omnivorum, Rhizoctonia solani, Botryis sp., Verticillium sp. and other soil fungi and plant materials.

For product identity, the applicant submitted adequate product chemistry data except for metabolite characterization and storage stability. Three studies were conducted to address the acute dermal, acute intraperitoneal injection and primary dermal irritation data requirements for the technical. Requests to waive testing for other Tier 1 acute human health toxicology data requirements were based on the results of these studies and published information. The EPA accepted the rationales for acute oral, acute pulmonary toxicity and infectivity and pathogenicity testing for the technical product as well as the acute oral, primary eye irritation. Similar rationales were submitted to waive testing for the acute toxicology data requirements for the EP and considered adequate to support its proposed registration. A cell culture test was not required for this fungal active ingredient.  The applicant reported that no hypersensitivity incidents occurred during research, development, or testing of Clonostachys rosea strain 321U. In light of the results of the acute toxicity/ pathogenicity data and information and the absence of hypersensitivity incidents, the EPA is not requiring testing at Tiers II and III (Appendix A).

The proposed use pattern is expected to limit exposure to nontarget organisms. The a.i. will be confined to dip tanks used for treatment, and is not expected to result in exposure under labeled use. Birds, wild mammals, and terrestrial plants are not expected to be exposed to Clonostachys rosea strain 321U in dip tanks, and would be expected to have very limited exposure to the a.i. that is present on freshly treated wood stored at the treatment facility. Nontarget insects and honey bees may be incidentally exposed to freshly treated lumber, though this exposure is also expected to be very limited. Aquatic organisms may be exposed if the treatment solution is released into the environment, such as with splash during treatment or wash-off of the treatment during rain events. Language is present on the label to limit exposure in aquatic environments (Appendix A).

Occupational Exposure
Despite the low toxicological profile of Clonostachys rosea strain 321U, the EPA is requiring baseline personal protective equipment (PPE) for handlers who, due to their occupation, may be exposed to the active ingredient for prolonged periods and/or numerous times. Handlers working with Clonostachys rosea strain 321U in pesticide applications are directed to wear a long-sleeved shirt, long pants, socks, shoes, waterproof gloves, protective eyewear and/or a dust/mist filtering respirator meeting National Institute for Occupational Safety and Health (NIOSH) standards of at least N-95, R-95, or P-95. For future products, the EPA may require additional/different PPE (e.g., chemical-resistant gloves), other than the standard described above, on a product-specific basis.  

Public comment
On October 1, 2009, EPA announced a new policy to provide a more meaningful opportunity for the public to participate in major registration decisions before they occur. According to this new policy, the Agency intends to provide a public comment period prior to making a registration decision for, at minimum, the following types of applications: new active ingredients; first food uses; first outdoor uses; first residential uses; and any registration decisions for which the Agency believes there may be substantial public interest.

Consistent with the new policy of making registration actions more transparent, Clonostachys rosea strain 321U is subject to a 15-day comment period as a "new active ingredient" whose registration would result in a "first outdoor use." While a final decision on registration is contingent upon review and consideration of public comments, the Agency presently believes that, based upon the preliminary risk assessment and information submitted in support of registration of Clonostachys rosea strain 321U, it is in the best interest of the public and the environment to issue the registration for Clonostachys rosea strain 321U. The basis for this preliminary decision can be found in the risk assessment for Clonostachys rosea strain 321U, which is characterized throughout this BRAD and the associated reference documents written by Agency scientists.  

II. ACTIVE INGREDIENT OVERVIEW

    Biological Name:  	Clonostachys rosea strain 321U previously known as Gliocladium roseum 321U. 
 
    Culture Deposit:	The MPCA is referred to as Clonostachys rosea strain 321U. It is stored in the Forintek Culture Collection and at the Centrallbureau voor schimmelcultures (BS 226.48) and the American Type Culture Collection (ATCC 10406).
    
 	OPP Chemical Code: 		021000	
 
	Type of Pesticide:				Microbial Pesticide  -  Fungicide

                              See Appendix B for specific information (e.g., use sites, application rates, methods of application, formulation types, and target pests) regarding the proposed labeling for pesticide products containing this active ingredient.

III. REGULATORY BACKGROUND

A.  Applications for Pesticide Product Registration

On March 25, 2012, William Stoneman, on behalf of AEF global, Inc., submitted applications for a manufacturing-use pesticide product, FPSolano(R) Technical 100% (EPA File Symbol: 89046-E), and an end-use pesticide product, FPSolano(R) (EPA File Symbol: 89046-R), to the U.S. Environmental Protection Agency (EPA) under section 3 of the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA). These pesticide products contain a new active ingredient, Clonostachys rosea strain 321U previously known as Gliocladium roseum strain 321U. 

Clonostachys rosea strain 321U was the subject of a North American Free Trade Agreement joint registration review conducted by the Environmental Protection Agency Office of Pesticide Programs and the Health Canada Pest Management Regulatory Agency. The PMRA documents for registration of Clonostachys rosea strain 321U are under review. For further information, see http://www.hc-sc.gc.ca/cps-spc/pest/part/consultations/index-eng.php. 

PMRA was the leader for the health effects reviews and EPA for the ecological reviews. PMRA informed the applicant of deficiencies July 27, 2012 (letter from Marion Law to Andreja Preradov). The applicant responded to the deficiencies to both PMRA and EPA on October 11, 2012.

On August 14, 2012 (77 FR 48519, EPA-HQ-OPP-2012-0507), the EPA announced receipt of these applications to register pesticide products containing a new active ingredient and opened a 30-day public comment period pursuant to the provisions of FIFRA section 3(c)(4). No comments were received following this publication when the 30 day comment period ended.                     

On March 5, 2013 PMRA informed the applicant of a second set of deficiencies arising out of the October 11, 2012 resubmission. The applicant responded on May 10, 2013. These data were subject to a second completness check and were found to be sufficient to proceed with full review.

The Pesticide Registration Improvement Renewal Act (PRIA II) date, originally scheduled for April 23, 2013, was renegotiated to April 30, 2014. One study, submitted to PMRA, was not part of EPA's data package. When it was submitted to EPA, the PRIA date was renegotiated to June 9, 2014 to allow time for its review and incorporation into the database for this MPCA.

B.	Food Tolerance Exemption 

No tolerance or exemption from the requirement of a tolerance is associated with or required for this regulatory action as only non-food uses are currently proposed for Clonostachys rosea strain 321U.  Accordingly, this action is outside the scope of the Food Quality Protection Act (FQPA). There are no CODEX maximum residue levels established for Clonostachys rosea strain 321U.

IV.  RISK ASSESSMENT

In the Federal Registers of October 26, 2007 (72 FR 60988) and August 30, 2012 (77 FR 52610), the EPA issued Final Rules on the data requirements to support registration of microbial pesticides and updated the definition for microbial pesticides. The data and information evaluated for this BRAD were considered in light of these requirements.

The classifications that are found for each data submission are assigned by Agency science reviewers and are an indication of the usefulness of the information contained in the documents for risk assessment. A rating of "ACCEPTABLE" indicates the study is scientifically sound and is useful for risk assessment. A "SUPPLEMENTAL" rating indicates the data provide some information that can be useful for risk assessment. The studies may have certain aspects determined not to be scientifically acceptable ("SUPPLEMENTAL: UPGRADABLE"). If a study is rated as "SUPPLEMENTAL: UPGRADABLE," the Agency always provides an indication of what is lacking or what can be provided to change the rating to "ACCEPTABLE." If there is simply a "SUPPLEMENTAL" rating, the reviewer will often state that the study is not required by the current 40 Code of Federal Regulations (CFR) part 158. Both "ACCEPTABLE" and "SUPPLEMENTAL" studies may be used in the risk assessment process, as appropriate. An "UNACCEPTABLE" rating indicates that new data need to be submitted.

For the acute toxicity data requirements, Toxicity Categories are assigned based on the hazard(s) identified from studies and/or other information submitted to the EPA in support of a pesticide registration. The active ingredient or particular product is classified into Toxicity Category I, II, III, or IV, where Toxicity Category I indicates the highest toxicity and Toxicity Category IV indicates the lowest toxicity.

A.  Product Analysis Assessment (40 C.F.R. § 158.2120)
   
The applicant fulfilled most of the product analysis data requirements for Clonostachys rosea strain 321U except for the storage stability data requirement. Acceptable studies include product chemistry and composition, analysis of samples, and physical and chemical characteristics. Storage stability of FPSolano(R) (manufactured at the AEF Global Inc. site) has been evaluated over a period of three months after storage at temperatures of -20°C, 4°C and 20°C using the method in accordance with both PMRA's and the EPA's microbial pesticides data requirements. The applicant has indicated that additional storage stability testing will be conducted at 6, 9, 12 and 24 months. These data must be submitted within 1 year of registration based on the potential that testing has been ongoing while the application was being processed.

The label for FPSolano(R) proposes storage conditions of 4°C for up to four months.  Based on the data submitted, FPSolano(R) continues to meet the guarantee after three months of storage at 4°C.  However, in the absence of data, the default storage period for wettable powder formulations is 6 months at 4°C. 

Refer to Tables 1 and 2 in Appendix A for a brief summary of the data requirements, including both generic and product-specific information.   
  
B.  Human Health Assessment (40 C.F.R. § 158.2140)

1.  Toxicity

All toxicology data requirements for non-food use of Clonostachys rosea strain 321U have been satisfied.

Tier II and Tier III studies were not required for Clonostachys rosea strain 321U based on the lack of acute toxicity/pathogenicity in the Tier I studies.

Refer to Tables 3 (generic - for Technical) and 4 (product specific - for EP) in Appendix A for a brief summary of the studies and information reviewed for the acute toxicology data requirements.  

	a.  	Endocrine Disruptors
	
As required by the Administrator under FFDCA section 408(p), the EPA has developed the Endocrine Disruptor Screening Program (EDSP) and has begun to implement the screening program that is to be used to test all pesticides in order to determine whether certain substances (including pesticide active and other ingredients) may have an effect in humans or wildlife similar to an effect produced by a "naturally occurring estrogen, or other such endocrine effects as the Administrator may designate." 

FFDCA section 408(p)(4) authorizes the Administrator, by order, to exempt from the requirements of the Estrogenic Substances Screening Program a biologic substance or other substance if a determination is made that the substance is not anticipated to produce any effect in humans similar to an effect produced by a naturally occurring estrogenic substance. 
Between October 2009 and February 2010, the EPA issued test orders/data call-ins for the first group of 67 chemicals, which contains 58 pesticide active ingredients and 9 inert ingredients. Clonostachys rosea strain 321U is not among the group of 58 pesticide active ingredients on the initial list to be screened under the EDSP. 

The EPA believes that Clonostachys rosea strain 321U, the active ingredient described in this document, likely is a substance that would not produce any effect in humans similar to an effect produced by a naturally occurring estrogenic substance. As such, pursuant to Section 408(p)(4), the EPA will determine in the future whether it can exempt Clonostachys rosea strain 321U from the requirements of the Section 408(p) EDSP. In the event the EPA does determine to exempt this substance from the EDSP, an order will be issued.  
For further information on the status of the EDSP, the policies and procedures, the list of 67 chemicals, the test guidelines and the Tier 1 screening battery, please visit our website at http://www.epa.gov/endo/.

2.  Federal Food, Drug, and Cosmetic Act (FFDCA) Considerations

No tolerance or exemption from the requirement of a tolerance is associated with or required for this regulatory action as only non-food uses are currently proposed for Clonostachys rosea strain 321U.  Accordingly, this action is outside the scope of the Food Quality Protection Act (FQPA). There are no CODEX maximum residue levels established for Clonostachys rosea strain 321U.

3.  Occupational Exposure and Risk Characterization

Handler exposure to Clonostachys rosea strain 321U is not expected to pose any undue risk. Regardless, the EPA is requiring appropriate personal protective equipment and precautionary statements on pesticide product labels to mitigate any potential risks to pesticide handlers due to prolonged and/or repeated exposures. Handlers working with Clonostachys rosea strain 321U for uses as labeled are directed to wear a long-sleeved shirt, long pants, socks, shoes, waterproof gloves, protective eyewear, and/or a dust/mist filtering respirator meeting NIOSH standards of at least N-95, R-95, or P-95. For future products, the EPA may require additional/different PPE (e.g., chemical-resistant gloves), other than the standard described above, on a product-specific basis.  

C.  Environmental Assessment (40 C.F.R. § 158.2150)

1.	Nontarget Organism Toxicology 

All nontarget toxicology data requirements for Clonostachys rosea strain 321U have been satisfied.  

Adequate Tier I nontarget organism data/information are available to support registration of Clonostachys rosea strain 321U.  The EPA found that guideline studies and scientific rationale submitted by the applicant are sufficient to fulfill the nontarget organism data requirements and for risk assessment purposes. Further testing of nontarget organisms at higher tier levels (i.e., Tiers II, III, and IV) is not required. The EPA performed an environmental risk assessment, based on information provided by the applicant, and determined that no effects are anticipated for nontarget species exposed to Clonostachys rosea strain 321U as a result of the proposed labeled applications.

Refer to Table 5 in Appendix A for a brief summary of the review of the data/information submitted for these data requirements.  

For a summary of the generic data requirements described in sections IV(C)(1), refer to Table 7 in Appendix A.

2.  Ecological Exposure and Risk Characterization  

Clonostachys rosea is a cosmopolitan microorganism. It is purported to be very common in soil and on organic matter; however, since it has not been the subject of much field research, field collections of this fungus are limited.  In Canada, isolates of this species that have been deposited in culture collections have been collected from British Columbia, Alberta, Ontario and Quebec. Moreover, Clonostachys rosea is common in a wide range of habitats in tropical, temperate, subarctic, and desert regions of the world. It was reported in cultivated, grassland, woodland, forest, heathland, freshwater, and coastal soils, particularly those of neutral to alkaline pH, and is not known to produce any adverse health or environmental effects (Pugh and Dickinson, 1965; Foley and Deacon, 1985; Sutton et al., 1997). 

3.  Environmental Fate Data 

As the information provided is sufficient to both satisfy the Tier I nontarget organism data requirements and for nontarget organism risk assessment for Clonostachys rosea strain 321U, further testing at higher tier levels (i.e., Tiers II, III, and IV) is not required.

4.  Threatened and Endangered Species Assessment

The EPA has determined that no effects are anticipated for nontarget species exposed to Clonostachys rosea strain 321U as a result of the proposed labeled applications. Therefore, a "No Effect" determination is made for direct and indirect effects to federally listed endangered and threatened ("listed") species and for their designated critical habitat. 

V.  ENVIRONMENTAL JUSTICE

The EPA seeks to achieve environmental justice -- the fair treatment and meaningful involvement of all people regardless of race, color, national origin, or income -- with respect to the development, implementation, and enforcement of environmental laws, regulations, and policies. Fair treatment means that no group of people, including racial, ethnic, or socioeconomic groups, should bear a disproportionate share of the negative environmental consequences resulting from industrial, municipal, and commercial operations or the execution of federal, state, local, and tribal environmental programs and policies. Meaningful involvement means that (1) potentially affected community residents have an appropriate opportunity to participate in decisions about a proposed activity that will affect their environment and/or health; (2) the public's contribution can influence the regulatory agency's decision; (3) the concerns of all participants involved will be considered in the decision-making process; and (4) the decision-makers seek out and facilitate the involvement of those potentially affected. The EPA has this goal for all communities and persons across the United States. 

To help address potential environmental justice issues, during the 15-day public participation comment period, the EPA sought information on any groups or segments of the population who, as a result their location, cultural practices, or other factors, may have atypical, unusually high exposure to Clonostachys rosea strain 321U, compared to the general population. The EPA received no public comments on this particular matter.

For additional information regarding environmental justice issues, please visit the EPA's website at http://www.epa.gov/compliance/environmentaljustice/index.html.

VI.  RISK MANAGEMENT DECISION

Section 3(c)(5) of FIFRA permits for the registration of a pesticide provided that all the following determinations are made:

	(1) Its composition is such as to warrant the proposed claims for it;
	(2) Its labeling and other material required to be submitted comply with the
	      requirements of FIFRA;
	(3) It will perform its intended function without unreasonable adverse effects on 
            the environment; AND
      (4) When used in accordance with widespread and commonly recognized 
            practice, it will not generally cause unreasonable adverse effects on the 
            environment.

To satisfy criterion 1, the EPA understands that Clonostachys rosea strain 321U is well-known to have fungicidal capabilities. The EPA has no knowledge that would contradict the claims made on the FPSolano(R)  biological fungicide labels, and the proposed pesticide products are not expected to cause unreasonable adverse effects on the environment when used according to their respective label instructions. Criterion 2 is satisfied by the product labels, as well as the data and information presented in this document. It is believed that the Clonostachys rosea strain 321U pesticide products will not cause any unreasonable adverse effects on the environment, and FPSolano(R)  biological fungicide (end-use pesticide product), in particular, is likely to control certain fungi as claimed, satisfying criterion 3. Criterion 4 is satisfied in that the Clonostachys rosea strain 321U pesticide products are not expected to cause unreasonable adverse effects when used according to label instructions. Therefore, the EPA believes FPSolano(R) Technical and FPSolano(R)  biological fungicide, containing the Clonostachys rosea strain 321U as a new active ingredient, are eligible for registration under FIFRA section 3(c)(5) for the labeled uses. 

VII.  ACTIONS REQUIRED OF THE REGISTRANT

A.  Final Printed Labeling

Before releasing pesticide products containing Clonostachys rosea strain 321U for shipment, the registrant is required to provide appropriate final printed labeling to the EPA.

B.  Terms of Registration

The following items will be terms of the FPSolano(R) Technical (EPA File Symbol: 89046-E) registration:

   1. Discussion of Formation of Unintentional Ingredients (OCSPP) Harmonized Guideline 885.1300:

     a) Confirmatory analysis for production of toxic secondary metabolites using the appropriate controls and samples, and information on the stability of potential toxins in the environment and at temperatures typical for kiln-drying.
   
     b) Microbial contaminant analysis and colony forming unit (CFU) count determination for a minimum of five batches of TGAI and EP produced at each of the alternative manufacturing and formulating sites
   
   2. Storage Stability (OCSPP Harmonized Guideline 830.6317): Within one year of registration, AEF Global, Inc., must provide the EPA with the results of a one-year storage stability study on FPSolano(R) Technical

The following items will be terms of the FPSolano(R) (EPA File Symbol: 89046-R) registration: 

   1. Storage Stability (Harmonized Guideline 830.6317): Within one year of registration, AEF Global, Inc., must provide the EPA with the results of a one-year storage stability study on FPSolano(R) if the applicant wishes to extend the length of storage time for FPSolano[(R)].  The label for FPSolano[(R)] proposes storage conditions of 4°C for up to four months. The applicant has indicated that additional storage stability testing will be conducted at 6, 9, 12 and 24 months. Based on the assumption that storage stability studies have been ongoing while the application was being processed, these data must be submitted within one year of registration.
   

C.  Reporting of Adverse Effects and Hypersensitivity Incidents

Notwithstanding the information stated in the previous sections, it should be clearly understood that certain specific data are required to be reported to the EPA as a requirement for maintaining the federal registration for a pesticide product. A brief summary of these types of data are described below.

Reports of all incidents of adverse effects to the environment must be submitted to the EPA under the provisions stated in FIFRA section 6(a)(2). Additionally, all incidents of hypersensitivity (including both suspected and confirmed incidents) must be reported to the EPA under the provisions of 40 C.F.R. § 158.2140(d).

VIII.  GLOSSARY OF ACRONYMS AND ABBREVIATIONS

A/cm[2]	acre per square centimeter
a.i.	active ingredient
BRAD	Biopesticides Registration Action Document
C.F.R.	Code of Federal Regulations
cfu	colony-forming units
cfu/kg	colony-forming units per kilogram
cfu/mL	colony-forming units per milliliter
cm	centimeter
EC	environmental concentration
EC50	half maximal effective concentration 
EDSP	Endocrine Disruptor Screening Program
EP	end-use pesticide product
EPA	United States Environmental Protection Agency 
FFDCA	Federal Food, Drug, and Cosmetic Act
FIFRA	Federal Insecticide, Fungicide, and Rodenticide Act
FR	Federal Register
g/A	grams per acre
g/lb	grams per pound
g/m[2]	grams per square meter
lb/A	pounds per acre
lb/ft[3]	pounds per cubic foot
LC50	median lethal concentration. A statistically derived concentration of a substance that can be 
           expected to cause death in 50% of test animals. It is usually expressed as the weight of substance per weight or volume of water, air, or feed (e.g., mg/L, mg/kg, or ppm).
LD50	median lethal dose. A statistically derived single dose that can be expected to cause death 
	in 50% of the test animals when administered by the route indicated (oral, dermal, or 
	inhalation). It is expressed as a weight of substance per unit weight of animal (e.g., mg/kg).
μg/μL	micrograms per microliter
μg/mL	micrograms per milliliter
mg/cm[2]	milligrams per square centimeter
mg/kg	milligrams per kilogram
mg/L	milligrams per liter
mg/mL	milligrams per milliliter
mL/L	milliliters per liter
mm	millimeter
MP	manufacturing-use pesticide product
MPCA	microbial pest control agent
MRID No.	Master Record Identification Number
ng	nanogram
NIOSH 	National Institute for Occupational Safety and Health
NOEC	no observed effect concentration
OPP	Office of Pesticide Programs
PC Code	Pesticide Chemical Code
PPE	personal protective equipment
ppm	parts per million
QC	quality control
TGAI 	technical grade of the active ingredient
U.S. 		   United States
USDA		   United States Department of Agriculture
USFWS	   United States Fish and Wildlife Service

IX.  BIBLIOGRAPHY

A.  Studies Submitted to Support the Clonostachys rosea strain 321U, previously known as Gliocladium roseum strain 321U, Pesticide Product Registrations

                                       
                                       
                                     MRID
                              Citation Reference

   48782601
Preradov, A. (2012) FPSolano(TM) Gliocladium roseum Strain 321U: Product Characterization and Analysis. Unpublished study prepared by AEF Global, Inc. 78p.

   48782602
Preradov, A. (2012) FPSolano(TM) Glicoladium roseum Strain 321U: Characterisation of the MPCA (A.I) Product Identity, Chemical Identity. Unpublished study prepared by AEF Global, Inc. 17p.

   48782603
Preradov, A. (2012) FPSolano(TM) Gliocladium roseum Strain 321U: Manufacturing Methods and Quality Assurance, Preservation and Maintenance of Production Strain. Unpublished study prepared by AEF Global, Inc. 23p.

   48782604
Preradov, A. (2012) FPSolano(TM) Gliocladium roseum Strain 321U: Storage Stability. Unpublished study prepared by AEF Global, Inc. 7p.

   48782605
Preradov, A. (2012) FPSolano(TM) Gliocladium roseum Strain 321U: Fulfillment of Data Requirement with Public Literature and/or Scientific Rationale: Acute Oral Toxicity/Pathogenicity. Unpublished study prepared by AEF Global, Inc. 9p.

   48782606
Preradov, A. (2012) FPSolano(TM) Gliocladium roseum Strain 321U: Fulfillment of Data Requirement with Public Literature and/or Scientific Rationale: Acute Pulmonary Infectivity and Toxicity. Unpublished study prepared by AEF Global, Inc. 10p.

   48782607
Monds, K. (2010) FPSolano(TM) Gliocladium roseum Strain 321U: Intraperitoneal (IP) Injection Toxicity/Pathogenicity Study: Final Report. Project Number: 13910/10. Unpublished study prepared by Stillmeadow, Inc. 23p.

   48782608
Preradov, A. (2012) FPSolano(TM) Gliocladium roseum Strain 321U: Fulfillment of Data Requirement with Public Literature and/or Scientific Rationale Hypersensitivity Incidents. Unpublished study prepared by AEF Global, Inc. 5p.

   48782609
Younger, C. (2010) FPSolano(TM) Gliocladium roseum Strain 321U: Acute Oral Toxicity Study in Bobwhite Quail: Final Report. Project Number: 13909/10. Unpublished study prepared by Stillmeadow, Inc. 19p.

   48782610
Preradov, A. (2012) FPSolano(TM) Gliocladium roseum Strain 321U: Fulfillment of Data Requirement with Public Literature and/or Scientific Rationale Wild Mammal Testing, Tier I. Unpublished study prepared by AEF Global, Inc. 8p.

   48782611
Brunner, S. (2010) FPSolano(TM) Gliocladium roseum: Microbial Pest Control Agent (MPCA) Freshwater Fish Test Oncorhynchus mykiss (Rainbow Trout): Final Report. Project Number: 13946/10. Unpublished study prepared by Stillmeadow, Inc. 15p.

   48782612
Brunner, S. (2010) FPSolano(TM) Gliocladium roseum: Microbial Pest Control Agent (MPCA) Freshwater Aquatic Invertebrate Test with Daphnia magna: Final Report. Project Number: 13947/10. Unpublished study prepared by Stillmeadow, Inc. 18p.

   48782613
Preradov, A. (2012) FPSolano(TM) Gliocladium roseum Strain 321U: Fulfillment of Data Requirement with Public Literature and/or Scientific Rationale Nontarget Aquatic Plants. Unpublished study prepared by AEF Global, Inc. 7p.

   48782614
Preradov, A. (2012) FPSolano(TM) Gliocladium roseum Strain 321U: Fulfillment of Data Requirement with Public Literature and/or Scientific Rationale Terrestrial Plants. Unpublished study prepared by AEF Global, Inc. 7p.

   48782615
Preradov, A. (2012) FPSolano(TM) Gliocladium roseum Strain 321U: Fulfillment of Data Requirement with Public Literature and/or Scientific Rationale: Terrestrial Arthropods. Unpublished study prepared by AEF Global, Inc. 6p.

   48782616
Preradov, A. (2012) FPSolano(TM) Gliocladium roseum Strain 321U: Fulfillment of Data Requirement with Public Literature and/or Scientific Rationale: Acute Oral Toxicity - Rat, Acute Inhalation Toxicity - Rat. Unpublished study prepared by AEF Global, Inc. 10p.

   48782617
Kuhn, J. (2010) FPSolano(TM) Gliocladium roseum: Acute Dermal Toxicity in Rats: Final Report. Project Number: 13954/10. Unpublished study prepared by Stillmeadow, Inc. 11p.

   48782618
Preradov, A. (2012) FPSolano(TM) Gliocladium roseum Strain 321U: Fulfillment of Data Requirement with Public Literature and/or Scientific Rationale: Primary Eye Irritation. Unpublished study prepared by AEF Global, Inc. 11p.

   48782619
Kuhn, J. (2010) FPSolano(TM) Gliocladium roseum: Acute Dermal Irritation Study in Rabbits: Final Report. Project Number: 13955/10. Unpublished study prepared by Stillmeadow, Inc. 11p.

   48782620
Stoneman, W. (2012) FPSolano(TM) Gliocladium roseum Strain 321U: Reference Volume for Fulfillment of Data Requirement with Public Literature and/or Scientific Rationale. Project Number: 13955/10. Unpublished study prepared by AEF Global, Inc. 158p.

   48978900
AEF Global Inc. (2012) Submission of Pesticide Use Data in Support of the Registration of FPSolano(TM). Transmittal of 1 Study.

   48978901
Bourassa, A. (2012) FPSolano(TM): Gliocladium roseum Strain 321U: Response to Deficiency Letter Sent on July 27, 2012 by Mario Law, PMRA. Project Number: 101/1094 2012/1160. Unpublished study prepared by AEF Global Inc. 255p.

   49172001
Bourassa, A. (2013) FPSolano and FPSolano Technical: Gliocladium roseum Strain 321U: Response to Deficiency Letter Sent on March 5, 2013 by Marion Law, PMRA. Unpublished study prepared by AEF Global, Inc. 1381 p.

   49351601

Brummer, S. (2011) Gliocladium roseum. FINAL REPORT: Microbial Pest Control Agent (MCPA): Freshwater Aquatic Invertebrate Test with Daphnia magna. Project Number: 14415/10. Unpublished study prepared by Stillmeadow, Inc. 28p.

                                       

B.  Environmental Protection Agency Risk Assessment Memoranda

U.S. EPA. 2014. Clonostachys rosea strain 321U. Memorandum from S. Borges, to S. Bacchus, dated May 13, 2014
 
C.  Other References

USDA. 1994. Extension Service, National Agricultural Pesticide Impact Assessment Program.

U.S. EPA. 2012b. Chapter 7 of the Label Review Manual (Precautionary Statements). Available from 
      http://www.epa.gov/oppfead1/labeling/lrm/chap-07.pdf.

U.S. EPA. 1996. Microbial Pesticide Test Guidelines  -  Background for Residue Analysis of Microbial 
      Pest Control Agents (OPPTS 885.2000). Available from 
      http://www.epa.gov/ocspp/pubs/frs/publications/Test_Guidelines/series885.htm.

USFWS. 2013. Endangered Species. Available from http://www.fws.gov/endangered/.

World Health Organization. 2011. Guidelines for Drinking-water Quality (Fourth Edition). Available 
      from http://whqlibdoc.who.int/publications/2011/9789241548151_eng.pdf.

             APPENDIX A.  MICROBIAL PESTICIDES DATA REQUIREMENTS 
                        (40 .F.R. PART 158  -  SUBPART V)
   A. Product Analysis

The applicant fulfilled most of the product analysis data requirements (except for the storage stability data requirement) for Clonostachys rosea strain 321U, including product chemistry and composition, analysis of samples, and physical and chemical characteristics, by acceptable guideline studies (Tables 1 and 2, Appendix A). 

The original submission package referred to the MPCA as Gliocladium roseum strain 321U.  Gliocladium roseum was reclassified as Clonostachys rosea, because G. roseum differed from the type species of Gliocladium, G. penicillioides, in morphology, ecology, teleomorph, and DNA sequence data. Therefore, the MPCA is to be referred to as Clonostachys rosea strain 321U.

Gliocladium catenulatum has also been transferred to the genus Clonostachys and is known as C. rosea f. catenulata.  The relationship between Gliocladium and Clonostachys is therefore considered close.  Furthermore, the Gliocladium genus is closely related to the Trichoderma genus.  For example, G. virens has been reclassified as T. virens.  As a result, information on various Gliocladium and Trichoderma species are considered in this review. The MPCA is permanently stored in liquid nitrogen in the Forintek Culture Collection and has also been deposited at the Centrallbureau voor schimmelcultures (BS 226.48) and the American Type Culture Collection (ATCC 10406). 

The taxonomic designation of the microbial pest control agent (MPCA) is as follows (MRID 49172001):

Kingdom
Fungi
Phylum
Ascomycota
Class
Sordariomycetes
Order
Hypocreales
Family
Bionectriaceae
Genera
Clonostachys
Species
rosea
Strain 
321U

Product identity

Clonostachys rosea strain 321U can be distinguished from species belonging to the closely related Gliocladium genus by morphology.  Sequencing of the internal transcribed spacer (ITS) can be used to distinguish the active ingredient from other Clonostachys species and other strains of C. rosea.  
   
Unintentional Ingredient analysis

Results of microbial contaminant testing for five batches of TGAI and five batches of EP (manufactured at the AEF Global Inc. site) were provided (MRID 49172001). Analyses of the total Mesophiles, total coliforms, fecal coliforms, fecal Enterococci and Staphylococci were acceptable (<10 cfu/mL for the Technical and <10 cfu/g for the EP). Although testing for Salmonella and Shigella species is not routinely conducted as part of the manufacturing process/quality assurance protocol, testing was carried out on five batches of EP using Health Canada protocols MFHPB-20 and MFLP-25 that include an enrichment step (MRID 49172001). Results provided for five batches of the TGAI and EP demonstrated that they were absent.

These microbial contaminant analysis data submitted for batches of FPSolano produced at the primary manufacturing and formulating site were acceptable for the registration of the pesticides.  Additional microbial contaminant analysis data on batches produced at the secondary manufacturing and formulating sites will be required.
   
   B.	Biological Properties of the MPCA(s):	

      1. Natural Occurrence of the Microorganism: 	

Clonostachy rosea has a cosmopolitan distribution.  It is common in a wide range of habitats in tropical, temperate, sub-arctic and desert regions of the world.  It has been reported from cultivated grassland and woodland, forest, heathland, freshwater and coastal soils.  This fungus has been frequently associated with cysts of Heterodera sp., Globodera sp. and other nematodes in soil and with sclerotia of Sclerotnia sclerotorium, Phymatotrichum omnivorum, Rhizoctonia solani, Botryis sp., Verticillium sp. and other soil fungi and plant materials (MRID 49172001). 

      2.  Target Organisms:

The MPCA is intended as an antisapstain product for control of blue stain and mould on freshly cut lumber and logs in hardwoods and softwoods.  Target organisms on which the MPCA has been tested include Ophiostoma piceae 387E, Aureobasidium pullulans 132S, Ceratocystis adiposa 251E, Alternaria alternata 2G, Cladosporium sphaerospermum 806B, Hormonema dematioides 742D, Rhinocladiella atrovierens 135E, O. minus 864A, Phialophora botulispora 707C and O. piliferum 55B (MRID 48978901).

The end-use product, FPSolano, is mixed with an alkaline solution prior to the wood being treated.  The wood staining fungi are susceptible to alkaline conditions while the MPCA is unaffected by high pH.  The alkaline solution displaces wild moulds and staining fungi from taking up their natural ecological niches on sawn wood products and logs allowing the MPCA to become established before the pest species.  The MPCA metabolises simple, non-structural nutrients present in woody plant materials, thereby minimizing nutrients for staining fungi (MRID 48978901).

The antagonistic behaviour of C. rosea/G. roseum towards other fungi has also been attributed to mycoparasitism, substrate competition, enzymatic activity (e.g., β1-3 glucanases,, chitinases) and induced resistance.  Moreover, production of toxic secondary metabolites may also exert an antibiotic effect (MRID 49172001).  

      3. Host Range:

The host range of C. rosea includes Acacia, Acer, Beta, Buxus, Cucumis, Daucus, Fragaria, Liquidambar, Lycopersicon, Metrosideros, Phaseolus, Phoenix, Pinus, Poncirus, Psilotum, Quercus, Rhizophora,  Trifolium, citrus and glycine (MRID 48978901).

TABLE 1.  Summary of Product Analysis Data Submitted for FPSolano(TM) Technical [the Technical Grade of the Active Ingredient (TGAI)], Clonostachys rosea strain 321U and Its End-Use Pesticide Product (EP), FPSolano(R)  (40 C.F.R. § 158.2120)
                         Harmonized Guideline Number 
                               Data Requirement
                                    Results
                          MRID No. or Other Reference
                                       
                                       
                                     TGAI
                                      EP
                                       
                                       
                                   885.1100
                                       
                               Product Identity
Submitted data fulfill the requirement for product identity. 
                                       
Submitted data fulfill the requirement for product identity. 
(EP contains a minimum of 5 x 10[8] CFU/gram). 
                                   48782602
                                       
                                   885.1200
                                       
                             Manufacturing Process
Submitted data fulfill the requirement for manufacturing process. 
                                   48782603
                                   885.1250
     Deposition of a Sample in a Nationally Recognized Culture Collection
The MPCA is referred to as Clonostachys rosea strain 321U.  It is stored in the Forintek Culture Collection and at the Centrallbureau voor schimmelcultures (BS 226.48) and the American Type Culture Collection (ATCC 10406). 
                                      N/A
                                   48782602
                                       
                                   885.1300
             Discussion of Formation of Unintentional Ingredients
Submitted data fulfill the requirement for discussion of formation of unintentional ingredients. 
                                   48782603
                                   885.1400
                              Analysis of Samples
        Submitted data fulfill the requirement for analysis of samples.
                                   48782603
                                   885.1500
                            Certification of Limits
                                      N/A
Limits listed on the confidential statement of formula are adequate.
                                   48782603

TABLE 2. Physical and Chemical Characteristics for FPSolano(R) Technical  [the Technical Grade of the Active Ingredient (TGAI)], Clonostachys rosea strain 321U and Its End-Use Pesticide Product (EP), FPSolano(R) (40 C.F.R. § 158.2120)
                          Harmonized Guideline Number
                               Data Requirement
                                    Results
                          MRID No. or Other Reference
                                       
                                       
                                     TGAI
                                      EP
                                       
                                   830.6302
                                     Color
                                 Not reported
                                 Not reported
                                   48978901
                                   48782602
                                   830.6303
                                Physical State
                                    Liquid
                                Wettable powder
                                       
                                   830.6304
                                     Odor
                                     mild 
                                 Not reported
                                       
                                   830.6313
     Stability to Normal and Elevated Temperatures, Metals, and Metal Ions
                                      N/A

                                      N/A
                                   830.6317
                               Storage Stability
Based on the data submitted, FPSolano(R) continues to meet the guarantee after three months of storage at 4°C.  However, in the absence of data, the default storage period for wettable powder formulations is 6 months at 4°C.  The label for FPSolano proposes storage conditions of 4°C for up to four months. The applicant has indicated that additional storage stability testing will be conducted at 6, 9, 12 and 24 months. Based on the assumption that storage stability studies have been ongoing while the application was being processed, these data must be submitted within one year of registration.
                                   48782604
                                   48978901
                                   830.6319
                                  Miscibility
                                      N/A
                                       
                                      N/A
                                       
                                       
                                       
                                      N/A
                                   830.6320
                           Corrosion Characteristics
                                      N/A
                                      N/A
                                       
                                   830.7100
                                   Viscosity
                                      N/A
Not required because EP is not a liquid form of microbial pesticide (refer to test note #5 of 40 C.F.R. § 158.2120(d)). 
                                       
                                   830.7300
           Density/Relative Density/Bulk Density (Specific Gravity)
                                     0.43
Soluble in water
                                   48978901

   C. Acute mammalian toxicology

Tier I mammalian toxicology data/information are sufficient for the registration of Clonostachys rosea strain 321U (see Tables 3 and 4 below). Three studies were conducted for the acute dermal, acute intraperitoneal injection and primary dermal irritation data requirements for the technical. The infectivity/pathogenicity endpoints were not addressed by the studies conducted for the acute dermal and acute injection guidelines. The test substances used in these studies were not representative of the technical or the end-use product (i.e., low level of active ingredient and no formulation ingredients added). Furthermore, the test substances used in these studies were liquid formulations whereas the end-use product is a wettable powder which may pose a greater potential for eye irritation.

In lieu of conducting acute oral and acute pulmonary infectivity/toxicity studies, the applicant has provided waiver rationales based on the results of the three above-mentioned studies, in addition to the mouse IP batch analysis studies, a literature search, the practices and design features employed in anti-sapstain facilities and the nature of the formulation ingredient. A similar rationale, the findings of a review paper and the use of PPE were found acceptable to waive testing for the primary eye irritation study. Appropriate PPE is required to mitigate potential skin sensitization that may be due to substances produced by all microorganisms that can elicit allergic reactions after repeated exposure to high concentrations. 

Requests to waive toxicology testing for the EP were accepted based on the foregoing studies and rationales submitted. No hypersensitivity incidents, including immediate-type or delayed-type reactions of humans and domestic animals, occurred during research, development, or testing of Clonostachys rosea strain 321U. Any future hypersensitivity incidents must be reported to the EPA (refer to test note #3 of 40 C.F.R. § 158.2140(d)).

A cell culture study is not required because Clonostachys rosea strain 321U is not a virus (refer to test note #4 of 40 C.F.R. § 158.2140(d)).

TABLE 3.  Summary of Tier 1 Toxicology Data Submitted for  FPSolano(R) Technical  [the Technical Grade of the Active Ingredient (TGAI)], Clonostachys rosea strain 321U (40 C.F.R. § 158.2140)
               Harmonized Guideline Number and Data Requirement
                                       
                              Summary of review 
                                     MRID 
                                    Number
                                   885.3050
                               (waiver request)
                                       
                       Acute Oral Toxicity/Pathogenicity
(waiver request based on IP, dermal, mouse IP batch analysis and nature of formulation ingredient)
Waiver rationale based on the results of an acute IP injection, acute dermal toxicity, dermal irritation and mouse IP batch analysis studies, a literature search, the practices and design features employed in anti-sapstain facilities and the nature of the formulation ingredient.  In lieu of acute oral pathogenicity testing, the registrant submitted a scientific rationale to waive testing.  

No signs of toxicity were noted in rats acutely exposed by IP injection to a 5.0x10[6] CFU/animal dose of C. rosea strain 321U. Infectivity was not assessed in this study although no signs of pathogenicity were observed.  

Results from a mouse IP batch analysis were also submitted in which no signs of adverse effects were observed amongst mice dosed with at least 10[6] CFU of C. rosea strain 321U via IP injection. 

Clonostachys rosea strain 321U was not toxic or irritating via the dermal route although the test material used was lower in toxicity than both the TGAI and the EP.  

As well, there were no incidents of pathogenicity/infectivity or other deleterious effects to mammalian species due to Clonostachys or Gliocladium reported in the public scientific literature.  

There were a number of cases of infections or adverse effects in immunocompromised individuals due to species of the related genus Trichoderma.  Toxicological studies on toxins and enzymes associated with Trichoderma species used purified forms of these substances whose levels are unlikely to be encountered under the proposed manufacturing process of use scenario.  In addition to the toxicology database, the registrant also cited the design and practices employed at anti-sapstain facilities that are designed to minimize exposure.  The formulation ingredient does not pose a concern.  The EP is not intended for use on food or feed crops.

       WAIVER ACCEPTEDTOXICITY CATEGORY III - Classification: Acceptable
                                       
                                       
                                       
                                       
                                       
                                       
                                       
                                       
                                       
                                       
                                       
                                       
                                       
                                   48782605
                                       
                                       
                                   885.3100
                                (14-Day study)
                                       
                     Acute Dermal Toxicity/Pathogenicity 
LD50 > 5050 mg/kg bw; test material 1x10[6] cfu/ml is lower than technical (2x10[8]cfu/ml) or EP (5x10[8] cfu/g). 

Materials and methods:
Tested Sprague-Dawley Rat -5/sex, 24-hour dermal exposure, with C. rosea strain 321U (1.0x10[6] CFU/mL) at dose of 5050 mg/kg bw (4.82 mL/kg bw) and animals observed for 14 days. Observations for mortality and clinical/behavioural signs of toxicity made three times on day of dosing and at least once daily thereafter.  Individual body weights recorded just prior to dosing and on Days 7 and 14. 

Results:
There were no mortalities noted throughout the study period.  All animals appeared normal and gained weight.  There were no abnormalities upon necropsy. 

LOW TOXICITY- viability of test substance is lower than that of TGAI or EP and there were no formulation ingredients in the test substance.

              TOXICITY CATEGORY IV -Classification: Supplemental

48782617

                                   885.3150
                               (waiver request)
                                       
                    Acute Pulmonary Toxicity/Pathogenicity 
Waiver Rationale based on mouse IP (48782607); 4 hr dermal irritation study in rabbits (48782619); acute dermal (48782617); public literature, nature of ingredients in formulation and low exposure potential due to the application method. In lieu of acute oral pathogenicity testing, the registrant submitted a scientific rationale to waive testing.  

No signs of toxicity were noted in rats acutely exposed by IP injection to a 5.0x10[6] CFU/animal dose of C. rosea strain 321U.  Infectivity was not assessed in this study although no signs of pathogenicity were observed.  

Results from a mouse IP batch analysis were also submitted in which no signs of adverse effects were observed amongst mice dosed with at least 10[6] CFU of C. rosea strain 321U via IP injection. 

Clonostachys rosea strain 321U was not toxic or irritating via the dermal route although the test material used was lower in toxicity than both the TGAI and the EP.  As well, there were no incidents of pathogenicity/infectivity or other deleterious effects to mammalian species due to Clonostachys or Gliocladium reported in the public scientific literature.  

There were a number of cases of infections or adverse effects in immunocompromised individuals due to species of the related genus Trichoderma.  Toxicological studies on toxins and enzymes associated with Trichoderma species used purified forms of these substances whose levels are unlikely to be encountered under the proposed manufacturing process of use scenario.  In addition to the toxicology database, the registrant also cited the design and practices employed at anti-sapstain facilities that are designed to minimize exposure.  The formulation ingredient does not pose a concern.  

SUPPLEMENTAL as infectivity has not been assessed via any route of exposure and inhalation will be a primary source of exposure to occupational workers. Appropriate PPE is required to mitigate any potential for inhalation exposure. 

WAIVER ACCEPTED
             TOXICITY CATEGORY III - Classification: Supplemental
                                   48782606.
                                   885.3200
                                (21-Day study)
                    Acute Injection Toxicity/Pathogenicity
At an IP dose of 5.0 x 10[6] CFU/animal, 1 month old Sprague Dawley rats were observed for 21 days.  Infectivity was not assessed although no signs of pathogenicity were noted. Replacement study waived based an acute IP injection, acute dermal toxicity, dermal irritation and mouse IP batch analysis studies, a literature search, the practices and design features employed in anti-sapstain facilities and the nature of the formulation ingredient. 

Materials and methods:
1) C. rosea strain 321U (5x106 CFU/mL); (15/sex); IP injection; 1 mL/animal (equivalent to 5.0x10[6] CFU/animal); 3 animals sacrificed 1 hour after dosing to very dose administered
2) Inactivated test substance; (5/sex); IP injection; 1 mL/animal (inactivated equivalent of 5x106 CFU/animal)
3) Untreated control (5/sex)

Observations: Mortality and adverse effects observations were made three times on Day 0 and daily thereafter.  Body weights were measured on Day 0 prior to dosing and on Days 7, 14 and 21.  Necropsies were performed at the end of the study on animals in the treatment group.  Organ weights were recorded for all animals.

Results: Test substance does not appear to be toxic. Infectivity was not assessed although no signs of pathogenicity were observed. 
There were no mortalities at any point of the study.  All animals appeared normal throughout.  No adverse effects were attributed to the test substance.  No significant differences between groups in body weights, body weight gains or organ weights.  No observable abnormalities noted at necropsy.

              TOXICITY CATEGORY IV - Classification: Acceptable.
                                       
                                       
                                       
                                       
                                       
                                       
                                       
                                   48782607
                                   49172001

--------------------------------------------------------------------------------

--------------------------------------------------------------------------------

--------------------------------------------------------------------------------

--------------------------------------------------------------------------------

--------------------------------------------------------------------------------
870.2500
--------------------------------------------------------------------------------

--------------------------------------------------------------------------------
Primary Dermal Irritation
A dermal irritation study was conducted for the technical. This study is required for the EP for labeling purposes to determine appropriate PPE to mitigate exposure risk to workers.

Material and methods:
2 ♂; 1 ♀ New Zealand white Rabbit treated with 0.5 mL/animal Clonostachys rosea strain 321U (1.0x106 CFU/mL) for 4-hour dermal exposure. Erythema and edema formation and any other dermal defects or irritation were scored at 1, 24, 28 and 72 hours after unwrapping.

Results:
MIS=0/8; MAS=0/8.
This dose is less than the technical and EP is a wettable powder. Guideline requires testing with EP.

              TOXICITY CATEGORY IV - Classification: Supplemental
                                   48782619

                          Hypersensitivity incidents
The applicant reported that no hypersensitivity incidents, including immediate-type or delayed-type reactions of humans and domestic animals, occurred during research, development, or testing of the TGAI or MP. Any future hypersensitivity incidents must be reported to the EPA (refer to test note #3 of 40 C.F.R. § 158.2140(d)). 
                                   48782608
                                   885.3500
                                 Cell culture
Not required because Clonostachys rosea strain 321U is not a virus (refer to test note #4 of 40 C.F.R. § 158.2140(d)).
                                      N/A

Table 4 includes a summary of the reviews of the requests to waive testing for the Clonostachys rosea strain 321U End-Use Pesticide Product (EP), FPSolano(R) (40 C.F.R. § 158.2140)

TABLE 4.  Summary of Tier 1 Toxicology Data Submitted for  Clonostachys rosea strain 321U End-Use Pesticide Product (EP), FPSolano(R) (40 C.F.R. § 158.2140)
                               Data Requirement
                                       
                                    Results
                                  MRID Number
                                   870.1100
                               (waiver request)
                                       
                              Acute Oral Toxicity
Waiver rationale based on results of an acute IP injection, acute dermal toxicity, dermal irritation and mouse IP batch analysis studies, a literature search, the practices and design features employed in anti-sapstain facilities and the nature of the formulation ingredient.  FPSolano is not intended for use on food or feed crops (refer to test note #5 of 40 C.F.R. § 158.2140(d)).

WAIVER ACCEPTED
              TOXICITY CATEGORY III - Classification: Acceptable 
                                       
                                       
                                       
                                   48782616
                                   870.1200
                               (waiver request)
                                       
                             Acute Dermal Toxicity
Waiver rationale based on MRID 48782617; safety of ingredients in formulation; use of PPE and low exposure potential. LD50 > 5050 mg/kg bw; test material 1x10[6] cfu/ml is lower than technical (2x10[8]cfu/ml) or EP (5x10[8] cfu/g); low toxicity; limited exposure via application method; appropriate PPE required, (refer to test note #5 of 40 C.F.R. § 158.2140(d)).

WAIVER ACCEPTED
              TOXICITY CATEGORY IV - Classification: Supplemental
                                   48782617
                                   870.1300
                               (waiver request)
                                       
                           Acute Inhalation Toxicity
Based on pulmonary study, low exposure due to method of application in dip tanks outdoors, appropriate mask with NIOSH prefix N-95.

WAIVER ACCEPTED 
             TOXICITY CATEGORY III - Classification: Supplemental 
                                   48782616
                                   870.2400
                               (waiver request)
                             Acute Eye Irritation
Waiver rationale based on results the results of IP injection, dermal irritation, dermal toxicity and IP batch analysis studies, the findings of a review paper and the use of PPE; the label must include use of PPE for eye protection based on dermal irritation and dermal toxicity studies. 

WAIVER ACCEPTED
              TOXICITY CATEGORY III - Classification: Acceptable
                                   48782618
                                 Sensitization
A dermal sensitization study was not conducted.  All microorganisms are recognized as being able to produce substance that can elicit allergic reactions after repeated exposure to high concentrations. Labelling must include appropriate PPE for potential sensitizing effects of microbial products must be included in the label. 

               Potential sensitizer - Classification: Acceptable
                                   48782608
                                   870.2500
                               (waiver request)
                                       
                           Primary Dermal Irritation
In lieu of a dermal irritation study on the end-use product, FPSolano, the applicant provided a waiver rationale based on the nature of the formulation ingredient, the absence of toxin production by the active ingredient and the requirement for personal protective equipment to minimize dermal exposure.  A primary dermal irritation study was conducted for the technical at a dose less than the technical and EP is a wettable powder. Although the claim that the active ingredient does not produce toxins was determined to be inconclusive, the waiver request was considered to be acceptable based on the identity and general safety of the formulation ingredient and the PPE requirements. See 870.2500 in TGAI table 1; MRID 48782619). 

WAIVER ACCEPTED 
               TOXICITY CATEGORY IV - Classification: Acceptable
                                   48782619
                          Hypersensitivity incidents
Published literature reported two incidents related to Gliocladium that were not very clearly linked directly to this pesticide active ingredient.  The applicant reported that no hypersensitivity incidents, including immediate-type or delayed-type reactions of humans and domestic animals, occurred during research, development, or testing of the TGAI or MP. Any future hypersensitivity incidents must be reported to the EPA (refer to test note #3 of 40 C.F.R. § 158.2140(d)). 
                                   49172001

Tiers II and III studies were not required for of Clonostachys rosea strain 321U based on the assessment of the submissions for the fungus in the Tier I studies.
   D. Ecological Effects

      Environmental Exposure

The proposed use pattern is expected to limit exposure to nontarget organisms. The a.i. will be confined to dip tanks used for treatment, and is not expected to result in exposure under labeled use. Birds, wild mammals, and terrestrial plants are not expected to be exposed to C. rosea strain 321U in dip tanks, and would be expected to have very limited exposure to the a.i. that is present on freshly treated wood stored at the treatment facility. Nontarget insects and honey bees may be incidentally exposed to freshly treated lumber, though this exposure is also expected to be very limited. Aquatic organisms may be exposed if the treatment solution is released into the environment, such as with splash during treatment or wash-off of the treatment during rain events. Language is present on the label to limit exposure in aquatic environments.

      Summary of Nontarget Effects Data

Table 5 provides the status of the nontarget organism data requirements for the active ingredient, Clonostachys rosea strain 321U, required under 40 CFR§ 158.2150. AEF Global submitted guideline studies to address toxicity/pathogenicity in birds, freshwater fish, and freshwater invertebrates, and submitted scientific rationale to address toxicity/pathogenicity in wild mammals, nontarget insects and honey bees, and terrestrial and aquatic plants. Data Evaluation Records for these studies and rationale were completed through joint review with Canada's Pesticide Regulatory Management Agency (PMRA). EPA determined that the information provided is sufficient to satisfy the nontarget organism data requirements for this new a.i. and its proposed uses. Additional testing at higher tiers is not required for the proposed uses. 

Table 5. Summary of Tier 1 Ecological Effects data submitted to comply with requirements published in 40 CFR § 158.2150 to support the registration of products containing the a.i., Clonostachys rosea strain 321U. 

OCSPP Guideline Number
Data Requirement
Results
MRID No.
885.4050
Avian Oral Toxicity
The LD50 in Northern bobwhite (Colinus virginianus) was determined to be > 5mL/kg bw (of test material containing 10[6] cfu/mL). The study had several deficiencies, including viability of the test material, which was less than that of the technical product.

                         Classification: Supplemental
48782609
885.4150
Wild Mammal Toxicity/Pathogenicity
Acceptable scientific rationale indicates that adverse effects to mammals would not be expected. This rationale is sufficient for assessing risks to wild mammals. 

WAIVER ACCEPTED 
                          Classification: Acceptable
48782610
885.4100
Avian Inhalation Toxicity/Pathogenicity
Not required; the nature of the microbial pesticide does not indicate potential pathogenicity to birds.
N/A
885.4200
Freshwater Fish Toxicity/Pathogenicity
In a 30-day study, mortality was >50% in rainbow trout (Oncorhyncus mykiss) exposed to Clonostachys rosea strain 321U at 10[4] cfu/mL, and an LC50 could not be established. Use pattern and proposed label language is expected to minimize aquatic exposure.

WAIVER ACCEPTED 
                         Classification: Supplemental
48782611
885.4240
Freshwater Invertebrate Toxicity/Pathogenicity
In a 7-day study, mortality was >50% in Daphnia magna exposed to Clonostachys rosea strain 321U at nominal concentration of 10[8] cfu/mL (likely actually <10[6]), and an LC50 could not be established. Use pattern and proposed label language is expected to minimize aquatic exposure.
                         Classification: Supplemental
48782612
885.4240
Freshwater Invertebrate Toxicity/Pathogenicity
In a 21-day study, the EC50 for survival was determined to be 5.5 x 10[5] cfu/mL, the IC50 and NOEC for reproduction were 5.8 x 10[5] and 5.0 x 10[5] cfu/mL, respectively, and the IC50 for growth was determined to be > 5.0 x 10[5] cfu/mL. Identity, purity, viability, and stability of the test substance were not demonstrated.

                         Classification: Supplemental
49351601
885.4280
Estuarine/Marine Fish and Invertebrate Testing
Not required; significant exposure in marine or estuarine environments is not expected due to the use pattern.
                                      N/A
885.4340
885.4380
Nontarget Insect Testing
Honey Bee Testing
Scientific rationale was sufficient to show that exposure to nontarget insects and honey bees is expected to be minimal based on the intended use pattern and proposed label language.

WAIVER ACCEPTED 
                          Classification: Acceptable
48782615
885.4300
Nontarget Plant Testing
Scientific rationale was sufficient to show that adverse effects to terrestrial and aquatic plants are not expected due to the proposed use pattern and cited data showing a lack of effects in some terrestrial plants.

WAIVER ACCEPTED 
                          Classification: Acceptable
48782613
48782614
                             Tiers II, III, and IV
Not required for Clonostachys rosea based on the acceptability of the scientific rationale and studies provided for Tier I.

	Ecological Effects Risk Characterization 

	Birds and Mammals

Exposure to birds and mammals is expected to be minimal because the intended use pattern is limited to treatment of wood contained in dip tanks. The label also contains sufficient language to limit environmental release and exposure to birds and mammals. Studies that were submitted did not show toxicity or pathogenicity in these animals; however, the studies were determined to have limited use for risk assessment. Since exposure is expected to be minimal, adverse effects to birds and mammals are not expected as a result of the proposed use of C. rosea strain 321U, as labeled.

	Nontarget Insects and Honeybees

Data in the public literature indicate that Clonostachys rosea is a potential entomopathogen (Toledo et al. 2006, Vega et al. 2008).  Insects may be exposed if they contact treated wood; however, while most of this exposure is likely to be incidental, some insects may be attracted to freshly treated wood as a potential water source while it is still wet. Since freshly treated wood will be limited to these treatment facilities and not widespread in the environment, such exposure is expected to be minimal and to have little impact on nontarget insect and honey bee populations. Therefore, adverse effects to nontarget insects and honey bees are not expected. 

Rationale presented by AEF Global to address nontarget data requirements included a proposal to include a label statement requiring users to cover freshly treated wood. This statement was part of EPA's and PMRA's consideration in acceptance of this rationale. While exposure to nontarget insects and honey bees may be limited, requiring users to cover freshly treated wood would reduce exposure further and also any uncertainty about risk due to the potential for effects on insects. It is recommended to include this requirement on the label.

	Freshwater Fish and Invertebrates

Studies submitted in support of this registration indicated adverse effects on survival in freshwater fish and survival, reproduction, and growth in freshwater invertebrates with exposure to C. rosea strain 321U. Exposure in aquatic environments could occur through splashing during treatment, spills, washing of treatment equipment, or wash-off of treated materials during rain events. Such exposure is expected to be limited, since these events would only occur at the lumber yard where the wood is treated, and these sites are likely limited within any landscape. Therefore, adverse effects to freshwater fish and invertebrates are not expected due to minimal exposure. 

The label contains language that is expected to sufficiently limit exposure through these types of releases into the environment. Rationale submitted as a supplement to the freshwater fish and invertebrate studies provided information on clean-up of spills, including application of a 5% bleach solution for one hour to the spill location. While the label does acknowledge that spills and splash are the responsibility of the applicator, it is recommended that the label provide information to the user on proper clean-up of spills so that environmental exposure is minimized. Label language requiring freshly treated lumber to be covered was also proposed in this rationale, and was considered in EPA's and PMRA's acceptance of this rationale. This language is also recommended, since this would prevent wash-off of the C. rosea strain 321U. 

	Marine/Estuarine Fish and Invertebrates

Exposure in marine/estuarine environments is not expected to be significant. Use of the EP is limited to facilities that cut or chip wood, and the product is applied by use of a dip tank. Exposure in nearby aquatic areas is expected to be limited, and any exposure in marine or estuarine sites would be further minimized by additional dilution within these larger water bodies. Therefore, adverse effects are not expected in marine/estuarine fish or invertebrates as a result of the use of C. rosea strain 321U, as proposed.

	Terrestrial and Aquatic Plants

Exposure to terrestrial and aquatic plants is also expected to be limited as a result of the use pattern and limited exposure. Some data were also submitted showing no observed adverse effects in cabbage, lettuce, and tomato plants. Label statements that limit any potential run-off from treated lumber would further reduce exposure in aquatic environments. Based on the use pattern limiting exposure and expected lack of effects, risk to terrestrial and aquatic plants is not anticipated as a result of the proposed uses of C. rosea strain 321U, as labeled.

      Threatened and Endangered Species Assessment 

Since EPA has determined that no effects are anticipated for nontarget species exposed to Clonostachys rosea strain 321U as a result of the proposed labeled uses, effects to federally listed threatened and endangered species and their designated critical habitats are also not expected. Therefore, a "No Effect" determination is made for direct and indirect effects to federally listed threatened and endangered species and their designated critical habitats for the proposed use of Clonostachys rosea strain 321U. 

	Conclusions

Adverse effects to nontarget organisms are not expected as a result of the proposed use of C. rosea strain 321U. Rationale submitted to support the registrations of products containing this a.i. proposed label language requiring that users cover freshly treated wood to prevent exposure to nontarget organisms. This language is not on the latest proposed label for the EP, but is recommended for inclusion on the label reduce any potential exposure to nontarget aquatic organisms, insects, and honey bees and any uncertainty with these risk conclusions. The label should also include guidance on proper clean-up of C. rosea strain 321U that may spill or splash during treatment.

	References

Toledo, A.V., Virla, E., Humber, R.A., Paradell, S.L., Lopez Lastra, C.C.  2006. Journal of Invertebrate Pathology 92: 7-10.

Vega, F.E., Posada, F., Aime, M.C., Pava-Ripoll, M., Infante, F., and Rehner, S.A.  2008.  Entomopathogenic fungal endophytes. Biological Control 46: 72-82. 

                        APPENDIX B.  PESTICIDE PRODUCTS

                                EPA Reg. Number
                               Registration Name
                           Percent Active Ingredient
                               Formulation Type
                                  Use Site(s)
                           Method(s) of Application
                              Application Rate(s)
                                  Target Pest
                                    89046-2
                             FPSolano(R) Technical
                         100% (3.8 x 10[9]CFU/ gallon)
                         Technical liquid formulation
                                      N/A
                                    89046-1
                                  FPSolano(R)
                               (End-use Product)
                                      20%
                                5 x 10[8] CFU/
                                     gram 
                          End Use  -  Wettable powder
                          Sawn wood products and logs
Apply by diptank application technology. 
Do not tank mix with any pesticides.
Mix 7 ounces of FPSolano(R) per 26 US gallon of water containing 4% w/v sodium carbonate and 1% w/v sodium bicarbonate e.g. per 26 US gallon of water for example: add 7 ounces of FPSolano(R), 8.8 lbs of sodium carbonate and 2.2 lbs of sodium bicarbonate. Complete with up to per 26 US gallons of water to make final suspension containing 3.8x10[9] cfu/quart of Clonostachys rosea strain 321U.
                                       
Biological protectant for treatment of green lumber against staining fungi (e.g. blue stain fungi) and molds
		[1] Go to www.regulations.gov. Conduct a search for Docket ID Number EPA-HQ-OPP-2012-0507. 
		[2] See http://oaspub.epa.gov/apex/pesticides/f?p=PPLS:1.