BIOPESTICIDES REGISTRATION ACTION DOCUMENT



## Heat-killed Burkholderia spp. strain A396 Cells and Spent Fermentation Media

Pesticide Chemical (PC) Code: 006534

U.S. Environmental Protection Agency Office of Pesticide Programs

Biopesticides and Pollution Prevention Division

DRAFT February 4, 2014Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 2 of 33

Biopesticides Registration Action Document

TABLE OF CONTENTS

I. EXECUTIVE SUMMARY ............................

................................................... ............................................ 4

II. ACTIVE INGREDIENT OVERVIEW

.................................................. ................................................... .. 6

III. REGULATORY BACKGROUND .............................

................................................... .............................. 7

A. Applications for Pesticide Product Registration

.................................................. .......................... 7

B. Food Tolerance Exemption .......................

................................................... .................................... 7

IV. RISK ASSESSMENT ...................................

................................................... .............................................. 7

A. Product Analysis Assessment (40 CFR § 158.2120)

................................................... .................... 8

B. Human Health Assessment (40 CFR § 158.2140) ....

................................................... .................... 8

C. Environmental Assessment (40 CFR § 158.2150) ...

................................................... ................... 13

V. ENVIRONMENTAL JUSTICE ........................

................................................... .................................... 156

VI. RISK MANAGEMENT DECISION ....................

................................................... ................................ 167

VII. ACTIONS REQUIRED OF THE REGISTRANT .........

................................................... ....................... 17

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS ....

................................................... ............... 18

IX. BIBLIOGRAPHY ................................

................................................... ................................................... . 19

A. Studies Submitted to Support the Active Ingredie nt, Heat-killed Burkholderia spp. strain A396 cells and spent fermentation media , and Pesticide Products, MBI-206 TGAI and Venerate , Registrations ....... 19

B. Environmental Protection Agency Risk Assessment Memoranda ........................................ ..... 25

C. Other References ..............................

................................................... ........................................... 25

APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS

.................................................. ......... 26

APPENDIX B. PESTICIDE PRODUCT ....................

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Biopesticides Registration Action Document

BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

Office of Pesticide Programs (OPP) Biopesticides and Pollution Prevention Division

Microbial Pesticides Branch

Science Reviews

Joel Gagliardi, Ph.D. Product Analysis, H uman Health John L. Kough, Ph.D. Product Analysis, Human Health Gail Tomimatsu, Ph.D. Environmental Effects Shannon Borges Environmental Effects

Regulations

Kimberly Nesci Chief, Microbial Pesticides B ranch Michael Glikes Regulatory Action LeaderHeat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 4 of 33

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I. EXECUTIVE SUMMARY

Background

In August 2010, Marrone Bio Innovations, Inc., subm itted applications for a new end-use pesticide product, Venerate (EPA File Symbol 84059- RU), containing a new technical grade active ingredient (TGAI), Burkholderia spp. strain A396, and a new manufacturing product, MBI-206 TGAI (EPA File Symbol 84059-RG), to the Uni ted States Environmental Protection Agency (EPA) under section 3 of the Federal Insecti cide, Fungicide, and Rodenticide Act (FIFRA). Concurrently with this application, MBI fi led a petition for the exemption from the requirement of a tolerance for residues of Burkholderia spp. strain A396 in or on all agricultural commodities (Pesticide Petition 0F7778). While thes e applications were under review by the EPA, the applicant modified the formulation of the TGAI from a live microbe to one that is deactivated, now called heat-killed Burkholderia spp. strain A396 cells and spent fermentation

media.

Burkholderia spp. strain A396 is a bacterium and naturally occurring insecti cide. As an insecticide, it controls many foliar feeding pests, such as coleopteran pests: soft-bodied insect pests, such as aphids, mites and whiteflies; and ne matodes. The proposed manufacturing product (MP) containing this new TGAI is called “MBI-206 TG AI.” The proposed end-use product (EP) containing this active ingredient, heat-killed Burkholderia spp. strain A396 cells and spent fermentation media, is called “Venerate,” a biologi cal insecticide primarily applied as a foliar spray. The mode of action for this killed microbial pestic ide is explained by bioactive metabolites that have insecticidal activity. The proposed EP is inte nded for use in field applications, tree fruit and nut crops, ornamental shade and forestry trees, tre e farms and plantations, turf grown for seed and on sod farms and ornamental bedding plants and flowers through ground, chemigation and aerial equipment. Intended uses also include seed t reatments with the product before planting, as well as homes and gardens.

EPA scientists reviewed product analysis, toxicolog y, and nontarget organism data and information (40 CFR §§ 158.2120, 158.2140, and 158. 2150, respectively) submitted to support the registration of the proposed heat-killed Burkholderia spp. strain A396 cells and spent fermentation media pesticide product. They found th at, overall, such data and information are adequate for risk assessment purposes, fulfill the current microbial pesticide data requirements, and support registration of the proposed product un der FIFRA section 3(c)(5). Terms of the proposed registrations would require the manufactur er within one year after registration to submit additional data on the EP’s storage stabilit y, corrosion characteristics, and the metabolites produced by the TGAI, heat-killed Burkholderia spp. strain A396 cells and spent fermentation media, which are integral to its mode of action as an insecticide.

Product Analysis

For the purposes of FIFRA section 3(c)(5) registrat ion, the product analysis data requirements
for heat-killed Burkholderia spp. strain A396 cells and spent fermentation media, in cluding product chemistry and composition, analysis of samp les, physical and chemical characteristics, and additional data concerning the manufacturing pr ocess that inactivates the bacterium, were fulfilled by acceptable guideline studies.Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 5 of 33

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Toxicology

The applicant submitted adequate mammalian toxicolo gy data and information to support the pesticide product. Acute toxicity studies showed th at, at a single high dose, heat-killed Burkholderia spp. strain A396 cells and spent fermentation media, is not toxic. Moreover, the applicant reported that no hypersensitivity inciden ts occurred during research, development, or testing of heat-killed Burkholderia spp. strain A396 cells and spent fermentation media. In light of the results of the acute toxicity data, the abse nce of hypersensitivity incidents, and the deactivation of the test material, the EPA did not require testing at higher tiers (i.e., Tiers II and III).

Tolerance Exemption

The EPA is issuing an exemption for the requirement of a tolerance for heat-killed Burkholderia

spp. strain A396 cells and spent fermentation media to allow residues of the active ingredient in or on all agricultural commodities. The EPA conclud ed that there is reasonable certainty that no harm will result to the U.S. population, including infants and children, from aggregate exposure
to residues of heat-killed Burkholderia spp. strain A396 cells and spent fermentation media. Da ta and information submitted by the petitioner support the conclusion that no dietary risks are expected to occur as a result of the use of heat-ki lled Burkholderia spp. strain A396 cells and spent fermentation media as an active ingredient in pesticide products. The EPA concludes that there will not be significant exposure to heat-kill ed Burkholderia spp. strain A396 cells and spent fermentation media through food and drinking water. This conclusion is based on: (1) the proposed application methods of the end-use pestici de product, Venerate (foliar spray and/or seed directed; no aquatic applications); (2) the fi ltering effect of many particulate soil types; and (3) the conditions (e.g., filtration and pH adjustm ents) water is subjected to in wastewater treatment systems and drinking water facilities. Ba sed on its review of the acute oral toxicity data, the EPA determined that, even if the AI were present on food or in drinking water, human exposure to this bacterium would not result in unre asonable adverse effects.

Occupational Exposure

Despite the low toxicological profile of heat-kille d Burkholderia spp. strain A396 cells and spent fermentation media, the EPA has proposed requiring baseline personal protective equipment (PPE) for handlers that, due to their occupation, m ay be subject to prolonged or repeated exposure to the active ingredient. The proposed pro duct label directs handlers working with heatkilled Burkholderia spp. strain A396 cells and spent fermentation media in a gricultural settings to wear a long-sleeved shirt, long pants, waterproo f gloves, socks, shoes and a dust/mist filtering respirator meeting National Institute for Occupatio nal Safety and Health (NIOSH) standards of at least N-95, R-95, or P-95. For future products, EPA may require additional PPE such as protective eyewear on a product-specific basis.

Nontarget Organisms

The applicant submitted studies of nontarget organi sm testing for the proposed new active ingredient. 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 data and other information provided by the applicant, and determin ed that adverse effects to most nontarget organisms are not anticipated from the proposed pes ticidal uses of heat-killed Burkholderia spp.Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 6 of 33

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strain A396 cells and spent fermentation media. For honeybees and nontarget insects, however, the Agency does not have sufficient information to conclude that the heat-killed bacterium will not pose risks to all nontarget insect species beca use the studies the applicant submitted on honeybees and nontarget insects were classified as either “invalid/incomplete” or “unacceptable.” Hence, directions for use on the pr oposed EP label include precautions to protect honeybees and beneficial insects. EPA made a “No Ef fect” determination for direct and indirect effects to federally listed endangered and threaten ed (“listed”) mammals, birds, and freshwater and marine/estuarine animals and for their designat ed critical habitats resulting from propsed uses. However, because of the uncertainties associa ted with potential effects to nontarget insects and plants, EPA concluded that additional analyses would be necessary to make a determination for these species.

Public Participation

On October 1, 2009, the 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 policy, the EPA intends to provide a public comment period prior to making a registration decision for, at minimum, the following types of ap plications: new active ingredients; first food uses; first outdoor uses; first residential uses; o r any other registration actions for which the Agency believes there may be significant public int erest.

Consistent with the policy of making registration a ctions more transparent, the pesticide product containing heat-killed Burkholderia spp. strain A396 cells and spent fermentation media, a n ew active ingredient, is subject to a 15-day comment p eriod. In addition to containing a new active ingredient, the registration of the pesticide produ ct results in the first outdoor and food uses for
heat-killed Burkholderia spp. strain A396 cells and spent fermentation media. The docket identification number associated with these registr ation actions, and accessed through either http://www.regulations.gov or http://www.epa.gov/pesticides/regulating/registrati on-status.html

, is EPA-HQ-OPP-2011-0010. The following documents ar e available for comment in EPA-HQOPP-2011-0010: (1) draft Heat-killed Burkholderia spp. strain A396 cells and spent fermentation media Biopesticide Registration Action Document (BR AD); and (2) draft labels for the proposed manufacturing-use pesticide product, MBI-206 TGAI ( EPA File Symbol 84059-RG), and the proposed end-use pesticide product, Venerate (EPA F ile Symbol 84059-RU). The EPA is soliciting public comments, based upon the data and information submitted in support of MBI206 and Venerate, to determine whether it is approp riate to issue the proposed registrations. The basis for proposed decisions can be found in the ri sk assessment for heat-killed Burkholderia

spp. strain A396 cells and spent fermentation media, whi ch is characterized throughout this

BRAD.

II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Heat-killed Burkholderia spp. strain A396 cells and spent
fermentation media

Culture Deposit: USDA Agricultural Research Service Patent Culture
Collection: NRLL B-50319

OPP Chemical Code: 006534Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 7 of 33

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Type of Pesticide: Microbial Pesticide –Insecticide.

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

III. REGULATORY BACKGROUND

A. Applications for Pesticide Product Registration

In August 2010, Marrone Bio Innovations, Inc. (addr ess: 2121 Second Street, Suite B-107; Davis, California 95618) submitted an application t o register a manufacturing product, MBI-206 TGAI (EPA File Symbol 84059-RG) and an end-use pest icide product, Venerate (EPA File Symbol 84059-RU), under FIFRA section 3. On Februar y 2, 2011 (76 FR 5805 ), the EPA announced receipt of the applications to register p esticide products containing a new active ingredient, one for manufacturing into end-use prod ucts, and a second for use in field applications, tree fruit and nut crops, ornamental shade and forestry trees, tree farms and plantations, turf grown for seed and on sod farms a nd ornamental bedding plants and flowers through ground, chemigation and aerial equipment. I ntended uses also include seed treatments with the product before planting, as well as home a nd garden sites. The EPA opened a 30-day public comment period pursuant to the provisions of FIFRA section 3(c)(4). No comments were received following this publication.

B. Food Tolerance Exemption

Concurrent with its registration applications and u nder Federal Food, Drug, and Cosmetic Act (FFDCA) section 408(d), Marrone Bio Innovations, In c., submitted a petition for an exemption from the requirement of a tolerance for Burkholderia spp. strain A396 (Pesticide Petition (PP) 0F7778). In the Federal Register of February 25, 20 11 (76 FR 10584 ), the EPA announced that Marrone Bio Innovations, Inc., proposed an exemptio n from the requirement of a tolerance for residues of the microbial pesticide, Burkholderia spp. strain A396, in or on all raw agricultural crops, to include residues of Burkholderia spp. strain A396, and the Agency opened a 30-day comment period. No comments were received following this publication. In a Final Rule, the EPA is amending the proposed tolerance exemption to account for the deactivation of the microbe, now called heat-killed Burkholderia spp. strain A396 cells and spent fermentation
media.

IV. RISK ASSESSMENT

In the Federal Register of October 26, 2007 ( 72 FR 61002 ), the EPA issued a Final Rule on the data requirements to support registration of microb ial pesticides and updated the definition of microbial pesticides. The rule became effective on December 26, 2007. The data and information evaluated for this BRAD were considered in light of these requirements.

The EPA classifies each data submission with an ind ication of the usefulness of the information
contained in the documents for risk assessment. A r ating of “acceptable” indicates the study is scientifically sound and is useful for risk assessm ent. A “supplemental” rating indicates the data provide some information that can be useful for ris k assessment. The studies may have certainHeat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 8 of 33

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aspects determined not to be scientifically accepta ble (“supplemental: upgradeable”). If a study is rated as “supplemental: upgradeable,” EPA always provides an indication of what is lacking or what can be provided to change the rating to “accep table.” If there is simply a “supplemental” rating, the reviewer will often state that the stud y is not required by 40 CFR Part 158. Both “acceptable” and “supplemental” studies may be used in the risk assessment process as appropriate. An “unacceptable” rating indicates tha t new data must be submitted.

For the acute toxicity data requirements, Toxicity Categories are assigned based on the hazard(s) identified from studies and/or other info rmation submitted to the EPA in support of a pesticide registration. The EPA classifies the acti ve ingredient or particular product into Toxicity Category I, II, III, or IV, where Toxicity Category I indicates the highest toxicity and Toxicity Category IV indicates the lowest toxicity (see 40 CFR § 156.62 ).

A. Product Analysis Assessment ( 40 CFR § 158.2120 )

For purposes of registration under FIFRA section 3( c)(5), the EPA determined that all product analysis data requirements for heat-killed Burkholderia spp. strain A396 cells and spent fermentation media have been fulfilled. The applica nt submitted studies to support storage stability and corrosion characteristics for up to 6 months of use; however, 12 month and 18 month studies will be submitted as terms of registr ation. Refer to Tables 1, 2, 3, and 4 in Appendix A for a summary of the data requirements, including both generic and product-specific
information.

B. Human Health Assessment ( 40 CFR § 158.2140 )

1. Toxicity

All toxicology data requirements for heat-killed Burkholderia spp. strain A396 cells and spent
fermentation media have been fulfilled. Acceptable Tier I mammalian toxicology data and information support registration of the proposed ma nufacturing-use product, MBI-206 TGAI, and proposed end-use product, Venerate. Furthermore , Tier II and Tier III studies were not required for heat-killed Burkholderia spp. strain A396 cells and spent fermentation media because of the lack of acute toxicity/pathogenicity in the Tier I studies. For a summary of the toxicology data described in sections IV(B)(1)(a) a nd IV(B)(1)(b) (see pages 8–10), refer to
Table 5 in Appendix A

.

a. Acute Toxicity/Pathogenicity – Tier I

Acute Oral Toxicity/Pathogenicity – Rat (Harmonized Guideline 885.3050; Master Record Identification Number; Acute Pulmonary Toxicity/Pat hogenicity – Rat (Harmonized Guideline 885.3150; and Acute Injection Toxicity/Pathogenicit y (Intravenous) – Rat (Harmonized Guideline 885.3200 (MRID Nos. 482127-08, 482127-09, 488310-02, 489833-02 ) and Waiver Request (MRID No. 482127-18) : The EPA waived this data requirement due to heat killing of the active ingredient after fermentation and batch veri fication by enrichment; hence, no live cells are present in any products.

Hypersensitivity Incidents (Harmonized Guideline 88 5.3400) : The applicant reported that no hypersensitivity incidents, including immediate-typ e or delayed-type reactions of humans and domestic animals, occurred during research, develop ment, or testing of heat-killed BurkholderiaHeat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 9 of 33

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spp. strain A396 cells and spent fermentation media. Any future hypersensitivity incidents must be reported to EPA (refer to test note #3 of 40 CFR § 158.2140(d)).

Cell Culture (Harmonized Guideline 885.3500) : This study is not required because heat-killed Burkholderia spp. strain A396 cells and spent fermentation media is n ot a virus (refer to test note #4 of 40 CFR § 158.2140(d)).

Acute Oral Toxicity– Rat (Harmonized Guideline 870. 1100; Master Record Identification Number (MRID No 491737-01) : Researchers fed three 8 week-old female Sprague-D awley, albino rats a single dose of Venerate (94.46 %, und iluted test article, heat-killed Burkholderia

spp. strain A396 cells and spent fermentation media) at doses of 4.92 mL/kg, or 5000 mg/kg body weight (bw). After 14 days, they observed no d eaths or abnormalities. Based on the results of this study, heat-killed Burkholderia spp. strain A396 cells and spent fermentation media does not appear to be toxic and/or pathogenic in rats wh en dosed at approximately 5000 mg/kg bw. The EPA classified the acute oral toxicity/pathogen icity as Toxicity Category IV, the lowest toxicity category, and the Agency rated the study a s acceptable.

Acute Dermal Toxicity - Rat (Harmonized Guideline 8 70.1200; MRID No 491737-02) : Researchers treated ten 8 week-old Sprague-Dawley, albino rats (5 per sex) to Venerate at doses of 4.97 mL/kg, or 5,050 mg/kg body weight bw for 24 hours. They observed the animals one hour after exposure and at periodic intervals for 1 4 days. The researchers did not find signs of dermal irritation during any of these observations, and they did not observe any abnormalities
after the rats were sacrificed and dissected. The E PA classified the dermal toxicity as Toxicity Category IV, and the Agency rated the study as acce ptable.

Acute Inhalation Toxicity– Rat (Harmonized Guidelin e 870-1300; MRID No.491737-03) : Researchers exposed ten 8 week-old Sprague-Dawley, albino rats (5 per sex) nose only via the

inhalation route to Venerate in undiluted test arti cles for 4 hours at concentrations of 5.22 mg/L. None of the rats died after 14 days, and the resear chers observed no abnormalities during necropsies. The EPA classified the acute inhalation toxicity as Toxicity Category IV, the lowest toxicity category, and the Agency rated the study a s acceptable.

Acute Eye Irritation Study - Rabbit (Harmonized Gui deline 870.2400; MRID No 491737-04) : Researchers instilled 0.1ml of Venerate into the co njunctival sac of the right eye of 2 male and 2 female, 10 week-old albino rabbits for 24 hours. Th ey observed the animals for 3 days and rated the effects with a maximum average irritation score of 0.0, meaning that the formulation is nonirritating to the eye. The EPA classified the eye i rritation as Toxicity Category IV, and the Agency rated the study as acceptable.

Acute Dermal Irritation Study - Rabbit (Harmonized Guideline 870.2500; MRID No 491737-05): Researchers applied 0.5 mL of Venerate to the trunk s of 2 male and 1 female, 10 week-old albino rabbits and then covered the exposed skin fo r 4 hours. After observing the rabbits for 3 days, the researchers determined that there was no dermal irritation to the skin, based on a maximum irritation score of 0.0 at 24, 48 and 72 ho ur observations. The EPA classified the primary dermal irritation as Toxicity Category IV a nd rated the study as acceptable.Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 10 of 33

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b. Acute Toxicology and Subchronic Toxicity/Path ogenicity – Tier II;

ReproductiveFertility Effects, Carcinogenicity, Imm unotoxicity, and Infectivity/Pathogenicity Analysis – Tier III

Tier II and Tier III studies were not required for heat-killed Burkholderia spp . strain A396 cells and spent fermentation media, based on the lack of acute toxicity/pathogenicity in the Tier I
studies for heat-killed Burkholderia spp . strain A396 cells and spent fermentation media.

c. Endocrine Disruptors

As required by the Administrator under the Federal Food, Drug, and Cosmetic Act (FFDCA), 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 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 designat e.” FFDCA section 408(p)(4) authorizes the Administrator, by order, to exempt from the require ments of the Endocrine Disruptor Screening Program a biologic substance or other substance if a determination is made that the substance is anticipated to not produce any effect in humans sim ilar to an effect produced by a naturally occurring estrogenic substance. Between October 20 09 and February 2010, EPA issued test orders/data call-ins for the first group of 67 chem icals, which contains 58 pesticide active ingredients and 9 inert ingredients. Heat-killed Burkholderia spp. strain A396 cells and spent

fermentation media is not among the group of 58 pesticide active ingr edients on the initial list to
be screened under the EDSP.

The EPA believes that h eat-killed Burkholderia spp. strain A396 cells and spent fermentation media is a substance that would not produce any effect in humans similar to an effect produced by a naturally occurring estrogenic substance. As such, and pursuant to Section 408(p)(4), the EPA will determine in the future whether it can exempt h eat-killed Burkholderia sp. strain A396 cells and spent fermentation media from the requirements of the Section 408(p)(4) EDS P. In the event the EPA does determine to exempt this substance fro m 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 scree ning battery, please visit our website: http://www.epa.gov/endo/

.

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

Section 408(c)(2)(A)(i) of FFDCA allows the EPA to establish an exemption from the requirement for a tolerance (the legal limit for a pesticide chemical residue in or on a food) only if the EPA determines that the exemption is “safe.” Section 408(c)(2)(A)(ii) of FFDCA defines
“safe” to mean that “there is a reasonable certaint y that no harm will result from aggregate exposure to the pesticide chemical residue, includi ng all anticipated dietary exposures and all other exposures for which there is reliable informa tion.” This includes exposure through drinking water and in residential settings, but does not inc lude occupational exposure. Pursuant to section 408(c)(2)(B) of FFDCA, in establishing or maintaini ng in effect an exemption from the requirement of a tolerance, the EPA must take into account the factors set forth in section 408(b)(2)(C) of FFDCA, which require the EPA to giv e special consideration to exposure of infants and children to the pesticide chemical resi due in establishing a tolerance exemption, and to “ensure that there is a reasonable certainty tha t no harm will result to infants and children fromHeat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 11 of 33

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aggregate exposure to the pesticide chemical residu e....” Additionally, section 408(b)(2)(D) of FFDCA requires that the EPA consider “available inf ormation concerning the cumulative effects of [a particular pesticide's] . . . residues and ot her substances that have a common mechanism of toxicity.”

The EPA performs a number of analyses to determine the risks from aggregate exposure to pesticide residues. First, the Agency determines th e toxicity of a pesticide. Second, the EPA examines exposure to the pesticide through food, dr inking water, and through other exposures that occur as a result of pesticide use in resident ial settings.

Consistent with section 408(b)(2)(D) of FFDCA, the EPA has reviewed the available scientific data and other relevant information, and considered its validity, completeness, and reliability and the relationship of this information to human risk. The Agency also considered available information concerning the variability of the sensi tivities of major identifiable subgroups of consumers, including infants and children.

Based on the acute toxicity/pathogenicity data and information discussed previously and presented in Table 5 in Appendix A , the data required for a FFDCA risk assessment forheatkilled Burkholderia spp. strain A396 cells and spent fermentation media have been fulfil led.

a. Aggregate Exposure

In examining aggregate exposure, section 408 of FFD CA directs EPA to consider available information concerning exposures from the pesticide residue in food and all other nonoccupational exposures, including drinking water from ground water or surface water and exposure through pesticide use in gardens, lawns or buildings (residential and other indoor uses).

Food Exposure and Risk Characterization : The EPA found that dietary exposure to heat-kille d Burkholderia spp. strain A396 cells and spent fermentation media, a n aturally occurring soil bacterium, is anticipated to be negligible. Due to the ubiquit ous nature of the organism and the concentrations of other closely related Burkholderia species that already exist in the environment, the likelihood of increased risk to hu mans or animals from the use of Venerate is
low.

Drinking Water Exposure and Risk Characterization : Exposure to residues of heat-killed Burkholderia spp . strain A396 cells and spent fermentation media in consumed drinking water is unlikely. The use patterns for Venerate primarily a re foliar, thereby limiting contact with surface water by drift and runoff. Furthermore, ground wate r is not expected to have significant exposure
to heat-killed Burkholderia spp. strain A396 cells and spent fermentation media sinc e, like other microorganisms, this microbial pesticide would like ly be filtered out by the particulate nature of many soil types. If it were to be transferred to su rface or ground waters that are intended for eventual human consumption (e.g., through spray dri ft or runoff) and directed to wastewater treatment systems or drinking water facilities, it likely would not survive the conditions water is subjected to in such systems or facilities, includi ng chlorination, pH adjustments, filtration, and/or occasionally high temperatures (Centers for Disease Control and Prevention 2009; U.S. EPA 2004). In the remote likelihood that heat-kille d Burkholderia spp. strain A396 cells and spent fermentation media is present in drinking wat er (e.g., water not subject to treatment systems or facilities), its target pest specificity and available data indicate no toxicity and/or pathogenicity is likely to occur with any drinking water exposure to heat-killed BurkholderiaHeat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 12 of 33

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spp. strain A396 cells and spent fermentation media that results from pesticide applications made in accordance with good agricultural practices (see section IV(B)(1)(a) on pages 8–9 and Table 5

in Appendix A ).

Non-occupational, Residential Risk Characterization : Given heat-killed Burkholderia spp . strain A396 cells and spent fermentation media’s natural o ccurrence in the environment, the EPA determined that non-occupational exposure to the ba cterium likely already is occurring. Additional exposure to the microorganism, due to pe sticidal applications, is not expected because such exposures do not exceed EPA’s level of concern , particularly in light of available data that
demonstrate heat-killed Burkholderia spp. strain A396 cells and spent fermentation media is n ot toxic (acute dermal toxicity and acute pulmonary to xicity/pathogenicity), non-irritating (primary dermal irritation), and not pathogenic (acute pulmo nary toxicity/pathogenicity) (see section IV(B)(1)(a) on pages 8–9 and Table 5 in Appendix A ).

b. Cumulative Effects from Substances with a Com mon Mechanism of Toxicity

Section 408(b)(2)(D)(v) of FFDCA requires that, whe n considering whether to establish, modify, or revoke a tolerance exemption, the EPA consider “ available information concerning the cumulative effects of [a particular pesticide’s] . . . residues and other substances that have a common mechanism of toxicity.”

The EPA has not found heat-killed Burkholderia spp. strain A396 cells and spent fermentation media to share a common mechanism of toxicity with other substances, and heat-killed Burkholderia spp. strain A396 cells and spent fermentation media does not appear to produce a toxic metabolite against target pests. For the purp oses of the tolerance action, therefore, the EPA
has assumed that heat-killed Burkholderia spp. strain A396 cells and spent fermentation media does not have a common mechanism of toxicity with o ther substances. Thus, section 408(b)(2)(D)(v) of the FFDCA does not apply. For in formation regarding the EPA’s efforts to
determine which chemicals have a common mechanism o f toxicity and to evaluate the cumulative effects of such chemicals, see the EPA’s website at http://www.epa.gov/pesticides/cumulative

.

c. Determination of Safety for the United States Population, Infants and Children

In considering the establishment of a tolerance or tolerance exemption for a pesticide chemical residue, FFDCA section 408(b)(2)(C) provides that t he EPA shall assess the available information about consumption patterns among infant s and children, special susceptibility of infants and children to pesticide chemical residues , and the cumulative effects on infants and
children of the residues and other substances with a common mechanism of toxicity. In addition, FFDCA section 408(b)(2)(C) provides that the EPA sh all apply an additional tenfold (10X) margin of exposure (safety) for infants and childre n in the case of threshold effects to account for prenatal and postnatal toxicity and the completenes s of the database on toxicity and exposure unless the EPA determines that a different margin o f exposure (safety) will be safe for infants and children. This additional margin of exposure (s afety) is commonly referred to as the Food Quality Protection Act Safety Factor. In applying t his provision, the EPA either retains the
default value of 10X or uses a different additional safety factor when reliable data available to the EPA support the choice of a different factor.

Based on the acute toxicity and pathogenicity data/ information discussed in section IV(B)(1)(a)Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 13 of 33

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(see pages 8–9) and Table 5 in Appendix A , the EPA concludes that there are no threshold effects of concern to infants, children, or adults when heat-killed Burkholderia spp. strain A396 cells and spent fermentation media is used as label ed in accordance with good agricultural practices. As a result, the EPA concludes that no a dditional margin of exposure (safety) is necessary to protect infants and children and that not adding any additional margin of exposure (safety) will be safe for infants and children.

Moreover, based on the same data/information and EP A analysis as presented directly above, the Agency is able to conclude that there is a reasonab le certainty that no harm will result to the United States population, including infants and chi ldren, from aggregate exposure to the residues
of heat-killed Burkholderia spp. strain A396 cells and spent fermentation media when it is used—as labeled and in accordance with good agricul tural practices—as an insecticide. Such exposure includes all anticipated dietary exposures and all other exposures for which there is
reliable information. The EPA has arrived at this c onclusion because, considered collectively, the
data and information available on heat-killed Burkholderia spp. strain A396 cells and spent fermentation media do not demonstrate toxic or path ogenic potential to mammals, including
infants and children.

3. Occupational Exposure and Risk Characterization

The EPA does not expect handler exposure to heat-ki lled Burkholderia spp. strain A396 cells and spent fermentation media to pose any undue risk . Regardless, appropriate personal protective equipment (PPE) and precautionary statements are re quired on pesticide product labels to mitigate any potential risks to pesticide handlers due to prolonged or repeated exposures. Handlers applying Venerate in agricultural settings must wear a long-sleeved shirt, long pants, waterproof gloves, socks, shoes, and a dust/mist fi ltering respirator meeting National Institute for Occupational Safety and Health (NIOSH) standards of at least N-95, R-95, or P-95. For future products containing this microorganism as an active ingredient, the EPA may require additional PPE such as protective eyewear on a product-specifi c basis.

4. Human Health Risk Characterization

The EPA considered human exposure to heat-killed Burkholderia spp. strain A396 cells and spent fermentation media in light of the standard f or registration in FIFRA and the relevant safety factors in FFDCA. The Agency determined that no unreasonable adverse effects to the United States population in general, and to infants and children in particular, will result when
heat-killed Burkholderia spp. strain A396 cells and spent fermentation media is u sed in accordance with EPA-approved labeling.

C. Environmental Assessment ( 40 CFR § 158.2150 )

Data and other information (e.g., scientific litera ture) submitted by the applicant to support requests to waive nontarget organism testing for he at-killed Burkholderia spp. strain A396 cells and spent fermentation media are sufficient to fulf ill the relevant microbial pesticide data requirements and for risk assessment purposes. Furt her testing of nontarget organisms at higher tier levels (i.e., Tiers II, III, and IV) is not re quired.Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 14 of 33

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The EPA performed an environmental risk assessment based on data and other information provided by the applicant, and the Agency determine d that adverse effects to nontarget organisms are not anticipated from the pesticidal u ses of heat-killed Burkholderia spp. strain A396 cells and spent fermentation media. For honeyb ees and nontarget insects, however, the Agency has insufficient information to conclude tha t the heat-killed bacterium will not pose risks to all nontarget insect species because the studies the applicant submitted on honeybees and a few species of nontarget insects were classified as either “invalid/incomplete” or “unacceptable.” Hence, directions for use on the proposed EP label include precautions to protect honeybees and beneficial insects. As a term of the proposed regis tration, the applicant would be required to
submit additional information on adverse effects on nontarget insects and honeybees. Moreover,
EPA has made a “No Effect” determination for direct and indirect effects to federally listed endangered and threatened (“listed”) mammals, birds , and freshwater and marine/estuarine animals and for their designated critical habitats resulting from propsed uses. However, because of the uncertainties associated with potential effe cts to nontarget insects and plants, EPA has concluded that additional analyses would be necessa ry to make a determination for these species.

For a summary of the generic data requirements desc ribed in sections IV(C)(1) (see pages 14– 17), refer to Table 6 in Appendix A

.

1. Ecological Exposure and Risk Characterization

a. Terrestrial Animals and Plants

Birds (Harmonized Guidelines 885.4050 and 885.2100) and Mammals (Harmonized Guidelines 885.4150):

Because applications of the end use product contain ing heat-killed Burkholderia spp . strain A396 cells and spent fermentation media include foliar s prays and soil directed products, birds and wild mammals could be exposed. Such exposure is exp ected to occur through ingestion of food items that contact the active ingredient, or incide ntal ingestion of soil or drinking water (e.g., puddles) that contain pesticide residues, or contact with treated surface s. Birds and wild mammals were not adversely affected when exposed at high dosage rates, as demonstrated in bird studies (MRID 49173707), and adverse effects w ere not observed in rats (MRIDs 49173701, 49173702, and 49173703). Therefore, adverse effects to birds and wild mammals are not expected if they are exposed to heat-killed Burkholderia spp. strain A396 cells and spent fermentation media as a result of uses on the propo sed end product’s label.

Nontarget Insects (Harmonized Guideline 885.4340), Honey Bees (Harmonized Guideline 885.4380), and Plants (Harmonized Guideline 885.430 0) :

Nontarget insects and honey bees may be exposed if they contact surfaces, including plants and soil, treated with heat-killed Burkholderia spp. strain A396 cells and spent fermentation media, or if this active ingredient is present on food ite ms. The EPA classified studies submitted on nontarget insects as either Invalid/Incomplete or U nacceptable, honey bees studies as Unacceptable, and nontarget plants as Invalid/Incom plete (see Table 6 on pages 29-33 for MRIDs). These classifications resulted from limitat ions in the studies’ approaches and experimental designs. For example, the researchers observed insects for only 24 hours after exposure to the pesticide while insecticidal activi ty could reasonably be expected after 24 hours.Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 15 of 33

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Because these studies did not clearly demonstrate h azard to these nontarget organisms, the risks posed cannot be determined without uncertainty. The refore, the Agency is requiring precautionary and mitigating text on the proposed e nd-use product’s label to protect pollinating insects and beneficial insects used in IPM practice s and in organic agriculture, as well as other nontarget insects that might be present in treated and other exposed areas. Also, mandatory incident reporting for cases of severe phytotoxicit y (e.g. no plant recovery after leaf burn, chlorosis or leaf curling) or reduced seed emergenc e following planting of treated seed must be reported to the manufacturer, and the manufacturer is required to report these incidents to the EPA (FIFRA, Section 6 A(2)).

b. Aquatic Animals and Plants

Freshwater Fish (Harmonized Guideline 885.4200, Fre shwater Invertebrates (Harmonized Guideline 885.1010), Estuarine/Marine Fish and Inve rtebrates (Harmonized Guideline 885.4280) (MRID Nos. 49173709 and 49173708) :

Freshwater fish and invertebrates could be exposed to the heat-killed Burkholderia spp . strain A396 cells and spent fermentation media from its pr oposed use on cranberries or from other applications that runoff and drift into aquatic are as. Applications from foliar sprays, in particular, could result in some runoff and drift of the applie d heat-killed Burkholderia spp. strain A396 cells and spent fermentation media into aquatic are as, but the concentrations resulting from drift and runoff are expected to be a small fraction of t he amount applied. Studies conducted on rainbow trout and freshwater invertebrates exposed to the proposed end-use product were classified as Acceptable by EPA scientists. Therefo re, adverse effects to freshwater fish and invertebrates are not expected from proposed applic ations of heat-killed Burkholderia spp. strain A396 cells and spent fermentation media as labeled.

2. Environmental Fate Data

As the data and information provided are sufficient to fulfill the Tier I nontarget organism data requirements and allow for nontarget organism risk assessment for heat-killed Burkholderia spp. strain A396 cells and spent fermentation media, fur ther testing at higher tier levels (i.e., Tiers II, III, and IV) is not required.

3. Threatened and Endangered Species Assessment

The EPA determined that no effects are anticipated for mammalian, avian, freshwater and marine/estuarine fish and invertebrate species expo sed to the active ingredient, heat-killed Burkholderia spp. strain A396 and spent fermentation media in the pro posed end-use product, Venerate, as a result of the proposed applications. Therefore , a “No Effect” determination is made for direct and indirect effects to federally l isted endangered and threatened (“listed”) mammals, birds, and freshwater and marine/estuarine animals and for their designated critical habitats. Because of uncertainties and data gaps as sociated with nontarget insects and plants,
direct effects to individuals of listed insect and plant species and indirect effects to listed insect and plant species cannot be precluded. Additional a nalysis of the potential effects is necessary to make a determination for these species.Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 16 of 33

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V. ENVIRONMENTAL JUSTICE

The EPA seeks to achieve environmental justice—the fair treatment and meaningful involvement of all people regardless of race, color, national o rigin, or income—with respect to the development, implementation, and enforcement of env ironmental laws, regulations, and policies. Fair treatment means that no group of people, inclu ding racial, ethnic, or socioeconomic groups, should bear a disproportionate share of the negativ e environmental consequences resulting from industrial, municipal, and commercial operations or the execution of federal, state, local, and tribal environmental programs and policies. Meaning ful involvement means that (1) potentially affected community residents have an appropriate op portunity to participate in decisions about a proposed activity that will affect their environmen t 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 iss ues, the EPA sought information on any groups or segments of the population who, as a resu lt of their location, cultural practices, or other factors, may have atypical, unusually high exposure to heat-killed Burkholderia spp. strain A396 cells and spent fermentation media, compared to the general population. For additional information regarding environmental justice issues, please visit the EPA’s web site at http://www.epa.gov/compliance/environmentaljustice/ index.html

.

VI. RISK MANAGEMENT DECISION

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

(1) Its composition is such as to warrant the prop osed 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 com monly recognized practice, it will not generally cause unreaso nable adverse effects on the
environment.

To satisfy criterion 1, the proposed products conta ining heat-killed Burkholderia spp. strain A396 cells and spent fermentation media have labels with claims supported by the EPA’s review of submitting studies, data, and other information. The EPA has no knowledge that would
contradict the claims made on the MBI-206 TGAI and Venerate labels, and these proposed pesticide products are not expected to cause unreas onable adverse effects on the environment when used according to label instructions. Criterio n 2 is satisfied by the proposed product labels, as well as the data and information presented in th is document. The Agency believes that the proposed heat-killed Burkholderia spp. strain A396 cells and spent fermentation media pest icide products will not cause any unreasonable adverse effects on the environment. In fact, Venerate is likely to provide protection against many insect pe sts, satisfying criterion 3. Criterion 4 is
satisfied in that the heat-killed Burkholderia spp . strain A396 cells and spent fermentation media pesticide products are not expected to cause unreas onable adverse effects when used accordingHeat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 17 of 33

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to label instructions. Therefore, the proposed prod ucts, MBI-206 TGAI and Venerate, both containing heat-killed Burkholderia spp. strain A396 cells and spent fermentation media as a new active ingredient, are eligible for registratio n under FIFRA section 3(c)(5) for labeled uses.

VII. ACTIONS REQUIRED OF THE REGISTRANT

A. Final Printed Labeling

Before MBI-206 TGAI and Venerate— the proposed pest icide products containing heat-killed Burkholderia spp. strain A396 cells and spent fermentation media –may be released for shipment, the registrant will be required to provid e appropriate final printed labeling to the EPA.

B. Terms of Registration

As a term of the proposed MBI-206 TGAI and Venerate registrations, Marrone Bio Innovations, Inc., will be required to submit confirmatory infor mation on the products within six months of registration.

Additionally, as terms of the proposed Venerate reg istration, Marrone Bio Innovations, Inc., will be required to submit studies on:

• Storage Stability (OCSPP 830.6317) and Corrosion Ch aracteristics for one year and 18 months in duration (OCSPP 830.6320).

• Identity (e.g., IUPAC names and CAS numbers) of the bioactive metabolites produced by the heat-killed Burkholderia spp. strain A396 cells and spent fermentation media.

C. Reporting of Adverse Effects and Hypersensitivi ty Incidents

Notwithstanding the information stated in the previ ous sections, it should be clearly understood that certain specific data must be reported to the EPA as a requirement for maintaining the federal registration for a pesticide product. Repor ts 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 (in cluding both suspected and confirmed incidents) must be reported to EPA under the provisions of 40 CFR § 158.2140(d).Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 18 of 33

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VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

BRAD Biopesticides Registration Action Document CFR Code of Federal Regulations
cSt centistokes EDSP Endocrine Disruptor Screening Program EP end-use pesticide product EPA United States Environmental Protection Agency ( the “Agency”) FFDCA Federal Food, Drug, and Cosmetic Act FIFRA Federal Insecticide, Fungicide, and Rodentici de Act FR Federal Register g/mL grams per milliliter lb/gal pounds per gallon

LD 50 median lethal dose. A statistically derived single dose that can be expected
to cause death in 50% of the test animals when adm inistered by the route indicated (oral, dermal, or inhalation). It is exp ressed as a weight of substance per unit weight of animal (e.g., mg/kg). MBI Marrone Bio Innovations, Inc. mg/kg milligrams per kilogram MP manufacturing-use pesticide product
MRID No. Master Record Identification Number NIOSH National Institute for Occupational Safety a nd Health OPP Office of Pesticide Programs
PC Code Pesticide Chemical Code
PP Pesticide Petition PPE personal protective equipment TGAI technical grade of the active ingredient
U.S. United StatesHeat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 19 of 33

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IX. BIBLIOGRAPHY

A. Studies Submitted to Support the Active Ingredi ent, Heat-killed Burkholderia spp. strain A396 cells and spent fermentation media, and Pesticide Product Registrations, MBI206 TGAI and Venerate.

MRID No. Study Information

48212700 Marrone Bio Innovations (2010) Submission of Produc t Chemistry, Toxicity, and

Exposure and Risk Data in Support of the Applicatio n for Registration of MBI-206 and the Petition for Tolerance of Burkholderia sp. Strain A396 in Food Commodities.
Transmittal of 19 Studies.

48212701 Koivunen, M. (2010) Product Chemistry for MBI-206 T GAI. Unpublished study

prepared by Marrone Bio Innovations. 454 p.

48212702 Kuhn, J. (2009) MOI 010: Acute Oral Toxicity Study (UDP) in Rats. Project

Number: 12680/09. Unpublished study prepared by Sti llmeadow, Inc. 11 p.

48212703 Kuhn, J. (2009) MOI 010: Acute Dermal Toxicity Stud y in Rats: Final Report.

Project Number: 13037/09. Unpublished study prepare d by Stillmeadow, Inc. 12 p.

48212704 Younger, C. (2009) MOI 010: Acute Inhalation Toxici ty Study in Rats: Final Report.

Project Number: 13038/09. Unpublished study prepare d by Stillmeadow, Inc. 19 p.

48212705 Kuhn, J. (2009) MOI 010: Acute Eye Irritation Study in Rabbits: Final Report.

Project Number: 13039/09. Unpublished study prepare d by Stillmeadow, Inc. 16 p.

48212706 Kuhn, J. (2009) MOI 010: Acute Dermal Irritation St udy in Rabbits: Final Report.

Project Number: 13040/09. Unpublished study prepare d by Stillmeadow, Inc. 10 p.

48212707 Kuhn, J. (2009) MOI 010: Skin Sensitization Study i n Guinea Pigs: Final Report.

Project Number: 13041/09. Unpublished study prepare d by Stillmeadow, Inc. 16 p.

48212708 Monds, K. (2009) MOI 010 Burkholderia spp.: Acute I njection Toxicity / Infectivity

Study (in Rats): Final Report. Project Number: 1305 5/09. Unpublished study prepared by Stillmeadow, Inc. 19 p.

48212709 Monds, K. (2009) MOI 010: Acute Injection Toxicity/ Pathogenicity Pilot Study in

Mice: Final Report. Project Number: 13098/09. Unpub lished study prepared by Stillmeadow, Inc. 8 p.

48212710 Monds, K. (2009) MOI 010: Subcutaneous Mouse Safety Study: Final Report.

Project Number: 13099/09. Unpublished study prepare d by Stillmeadow, Inc. 10 p.

48212711 Younger, C. (2009) MOI 010: Acute Oral Toxicity Stu dy in Bobwhite Quail: Final

Report. Project Number: 13255/09. Unpublished study prepared by Stillmeadow, Inc. 15 p.

48212712 Hartwell, T. (2010) MOI 010: Microbial Pest Control Agent (MPCA) Freshwater

Fish Test with Pimephales promelas: Final Report. P roject Number: 13286/09. Unpublished study prepared by Stillmeadow, Inc. 19 p.

48212713 Hartwell, T. (2009) Microbial Pest Control Agent (M PCA) Freshwater Aquatic

Invertebrate Test with Daphnia magna: Final Report. Project Number: 13287/09. Unpublished study prepared by Stillmeadow, Inc. 18 p.Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 20 of 33

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48212714 Younger, C. (2010) MBI 010: Green Lacewing Microbia l Testing: Amended Final

Report. Project Number: 13257/09. Unpublished study prepared by Stillmeadow, Inc. 18 p.

48212715 Younger, C. (2010) MBI 010: Parasitic Wasp, Aphidiu s matricariae, Non-Target

Insect Microbial Testing: Final Report. Project Num ber: 13903/10. Unpublished study prepared by Stillmeadow, Inc. 14 p.

48212716 Younger, C. (2009) MOI 010: Ladybird Beetle Non-Tar get Insect Microbial Testing:

Final Report. Project Number: 13336/09. Unpublished study prepared by Stillmeadow, Inc. 12 p.

48212717 Younger, C. (2010) MBI 010: Honey Bee, Apis mellife ra, Non-Target Insect

Microbial Testing: Final Report. Project Number: 13 905/10. Unpublished study prepared by Stillmeadow, Inc. 12 p.

48212718 Pitts, K. (2010) Response to Tier 1 Microbial Pesti cide Data Requirements for MBI206 TGAI. Unpublished study prepared by Marrone Bio Innovations. 26 p.

48212719 Pitts, K. (2010) Endangered Species Evaluation for MBI-206. Unpublished study

prepared by Marrone Bio Innovations. 17 p.

48212800 Marrone Bio Innovations (2010) Submission of Produc t Chemistry, Fate, and

Toxicity Data in Support of the Application for Reg istration of MBI-206 EP.
Transmittal of 3 Studies.

48212801 Koivunen, M. (2010) Product Chemistry for MBI-206 E P. Unpublished study

prepared by Marrone Bio Innovations. 182 p.

48212802 Morgan, C. (2010) MBI-206 EP - Nontarget Plants. Pr oject Number: 5301/150.

Unpublished study prepared by Marrone Bio Innovatio ns. 14 p.

48212803 Pitts, K. (2010) Response to Tier 1 Microbial Pesti cide Data Requirements for MBI206 EP. Unpublished study prepared by Marrone Bio I nnovations. 122 p.

48625801 Brunner, S. (2011) MBI-206 EP: Rainbow Trout (Oncor hynchus mykiss) 96-Hour

Acute Toxicity Test: Final Report. Project Number: 14343/10. Unpublished study prepared by Stillmeadow, Inc. 13p.

48745801 Cordova-Kreylos, A. (2012) Amended MBI-206 Microbia l Identification Study of

MRID 48212701. Unpublished study prepared by Marron e Bio Innovations. 450p.

48745802 Cordova-Kreylos, A. (2012) Amended Literature Revie w of MRID 48212107: The

Genus Burkholderia with Relevance to Agricultural a nd Biotechnological Applications. Unpublished study prepared by Marrone Bio Innovations. 1231p.

48745803 Pitts, K. (2012) MBI-206 TGAI and MBI-206 EP: Test Substance Identification.

Unpublished study prepared by Marrone Bio Innovatio ns. 44p.

48745804 Younger, C. (2010) Parasitic Wasp, Aphidius matrica riae, Non-target Insect

Microbial Testing: Final Report. Project Number: 13 903/10. Unpublished study prepared by Stillmeadow, Inc. 14p.

48745805 Pitts, K. (2012) MBI-206 TGAI Response to Completen ess Check Clarification /

Deficiencies. Unpublished study prepared by Marrone Bio Innovations. 36p.Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 21 of 33

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48745901 Mileson, B. (2012) Response to Tier 1 Microbial Pes ticide Data Requirements for

MBI-206 EP. Unpublished study prepared by Marrone B io Innovations. 105p.

48745902 Pitts, K. (2012) MBI-206 EP: Response to Completene ss Check

Clarifications/Deficiencies. Unpublished study prep ared by Marrone Bio Innovations. 13p.

48768501 Lam, R. (2012) MBI-206 EP: Amended Product Chemistr y. Unpublished study

prepared by Marrone Bio Innovations. 79p.

48769201 Lam, R. (2012) MBI-206 TGAI: Amended Manufacturing Process Study.

Unpublished study prepared by Marrone Bio Innovatio ns. 58p.

48769202 Graham, P. (2012) MBI-206 TGAI: Ten Batch Analysis Study in Heat Killed MBI206 TGAI. Project Number: 8202/90. Unpublished stud y prepared by Marrone Bio Innovations. 21p.

48769203 Graham, P. (2012) MBI-206 TGAI: Time/Temperature St udy for the Inactivation of

(Burkholderia sp. Strain A396). Project Number: 820 2/162. Unpublished study prepared by Marrone Bio Innovations. 25p.

48831001 Pitts, K. (2012) MBI-206 TGAI: Response to Agency C ompleteness Check.

Unpublished study prepared by Marrone Bio Innovatio ns. 362p.

48831002 Mileson, B. (2012) Supplemental Response to Tier 1 Requirements for MBI-206

TGAI. Unpublished study prepared by Technology Scie nces Group, Inc. 9p.

48831101 Pitts, K. (2012) MBI-206 EP: Three Month Storage St ability and Corrosion

Characteristics Study. Unpublished study prepared b y Marrone Bio Innovations. 73p.

48983300 Marrone Bio Innovations (2012) Submission of Produc t Chemistry, Toxicity and

Pesticide Use Data in Support of the Registrations of MBI-206 TGAI and MBI-206
EP. Transmittal of 5 Studies.

48983301 Cordova-Kreylos, A. (2012) MBI-206 Deficiency Respo nse: Microbial Identification

Addendum to MRID Numbers 482127-01 and 487458-01. U npublished study prepared by Marrone Bio Innovations. 253p.

48983302 Doig, A. (2012) Validation of Test Organism Recover y. Project Number: 16420/12.

Unpublished study prepared by Stillmeadow, Inc. 13p

.

48983303 Waller, J. (2012) MBI-206 TGAI: Expanted Time/Tempe rature Study for the Killing

of Burkholderia sp. Strain A396. Project Number: 42 07. Unpublished study prepared by Marrone Bio Innovations. 22p.

48983304 Pitts, K. (2012) MBI-206: Standard Operating Proced ures. Project Number:

MBI/MB/2001/SOP, MBI/FM/2001/SOP, MBI/M/4002/SOP. U npublished study prepared by Marrone Bio Innovations. 76p.

48983305 Pitts, K. (2012) MBI-206 EP: Storage Stability and Corrosion Characteristics Study,

Amendment of MRID 4881101. Unpublished study prepar ed by Marrone Bio Innovations. 85p.

49028800 Marrone Bio Innovations (2012) Submission of Produc t Chemistry Data in Support

of the Application for Registration of MBI-206 EP. Transmittal of 1 Study.Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 22 of 33

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49028801 Waller, J. (2012) MBI-206 EP: Amended Manufacturing Process Study-December

2012. Unpublished study prepared by Marrone Bio Inn ovations. 8p.

49028900 Marrone Bio Innovations (2012) Submission of Produc t Chemistry Data in Support

of the Application for Registration of MBI-206 TGAI . Transmittal of 1 Study.

49028901 Waller, J. (2012) MBI-206 TGAI: Amended Manufacturi ng Process Study-December

2012. Unpublished study prepared by Marrone Bio Inn ovations. 19p.

49171900 Marrone Bio Innovations (2013) Submission of Toxici ty Data in Support of the

Application for Registration of MBI-206 TGAI. Trans mittal of 1 Study.

49171901 Bowen, R. (2013) MBI-206 Sterile Cell Lysate: Rever se Mutation in Five Histidinerequiring Strains of Salmonella typhimurium Using a Treat and Plate Methodology. Project Number: 8278780/OCR. Unpublished study prep ared by Covance Laboratories, Ltd. 37p.

49173700 Marrone Bio Innovations (2013) Submission of Toxici ty and Fate Data in Support of

the Application for Registration of MBI-206 EP. Tra nsmittal of 21 Studies.

49173701 Kuhn, J. (2013) MBI-206 EP: Acute Oral Toxicity (UD P) in Rats. Project Number:

17300/13/OCR. Unpublished study prepared by Stillme adow, Inc. 10p.

49173702 Kuhn, J. (2013) MBI-206 EP: Acute Dermal Toxicity i n Rats. Project Number:

17301/13/OCR. Unpublished study prepared by Stillme adow, Inc. 11p.

49173703 Doig, A. (2013) MBI-206 EP: Acute Inhalation Toxici ty in Rats. Project Number:

17302/13/OCR. Unpublished study prepared by Stillme adow, Inc. 17p.

49173704 Kuhn, J. (2013) MBI-206 EP: Acute Eye Irritation in Rabbits. Project Number:

17303/13/OCR. Unpublished study prepared by Stillme adow, Inc. 17p.

49173705 Kuhn, J. (2013) MBI-206 EP: Acute Dermal Irritation in Rabbits. Project Number:

17304/13/OCR. Unpublished study prepared by Stillme adow, Inc. 11p.

49173706 Kuhn, J. (2013) MBI-206 EP: Skin Sensitization in G uinea Pigs. Project Number:

17305/13/OCR. Unpublished study prepared by Stillme adow, Inc. 16p.

49173707 Younger, C. (2013) MBI-206 EP: Acute Oral Biochemic al Toxicity Limit Study in

Bobwhite Quail. Project Number: 17296/13/OCR. Unpub lished study prepared by Stillmeadow, Inc. 16p.

49173708 Mikulas, J. (2013) MBI-206 EP: Daphnia magna 48-Hou r Acute Toxicity Test.

Project Number: 17311/13/OCR. Unpublished study pre pared by Stillmeadow, Inc. 14p.

49173709 Mikulas, J. (2013) MBI-206 EP: Rainbow Trout (Oncor hynchus mykiss) 96-Hour

Acute Toxicity Test. Project Number: 17312/13/OCR. Unpublished study prepared by Stillmeadow, Inc. 12p.

49173710 Morgan, C. (2013) MBI-206 EP: Terrestrial Plant Tox icity, Vegetative Vigor. Project

Number: 1508/77/OCR. Unpublished study prepared by Marrone Bio Innovations. 21p.

49173711 Lawrence, J. (2013) Oral Limit Test of MBI-206 EP o n the Honey Bee

HYMENOPTERA: Apidae Apis mellifera) in a Laboratory Study in North Carolina:Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 23 of 33

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Final Report. Project Number: S13/02866/OCR. Unpubl ished study prepared by Eurofins-Agroscience Services, Inc. 32p.

49173712 Lawrence, J. (2013) Contact Limit Test of MBI-206EP on the Honey Bee

(HYMENOPTERA: Apidae Apis mellifera) in a Laborator y Study in North Carolina: Final Report. Project Number: S13/02867/OCR. Unpubl ished study prepared by Eurofins-Agroscience Services, Inc. 30p.

49173713 Lawrence, J. (2013) Dose Respnse Effects of MBI-203 DF and MBI-206 EP on the

Parasitic Wasp (HYMENOPTERA: Aphelinidae Encarsia f ormosa) in a Translational Study in North Carolina: Final Report. Project Numb er: S13/00307/OCR. Unpublished study prepared by Eurofins-Agroscience Services, Inc. 52p.

49173714 Lawrence, J. (2013) Dose Response Effects of MBI-20 3 DF and MBI-206 EP on the

Rove beetle (COLEOPTERA: Staphylinidae Atheta coria ria) in a Translational Study in North Carolina: Final Report. Project Number: S1 3/00301/OCR. Unpublished study prepared by Eurofins-Agroscience Services, In c. 58p.

49173715 Lawrence, J. (2013) Dose Response Effects of MBI-20 3 DF and MBI-206 EP on the

Bumble Bee (HYMENOPTERA: Apidae Bombus sp.) in a Tr anslational Study in North Carolina: Final Report. Project Number: S13/0 0300/OCR. Unpublished study prepared by Eurofins-Agroscience Services, Inc. 53p

.

49173716 Lawrence, J. (2013) Dose Response Effects of MBI-20 3 DF and MBI-206 EP on the

Minute Pirate Bug (HEMIPTERA: Anthocoridae Orius in sidiosus) in a Translational Study in North Carolina: Final Report. Project Numb er: S13/00302/OCR. Unpublished study prepared by Eurofins-Agroscience Services, Inc. 55p.

49173717 Lawrence, J. (2013) Dose Response Effects of MBI-20 3 DF and MBI-206 EP on the

Parasitic Wasp (HYMENOPTERA: Aphelinidae Aphidius c olemani) in a Translational Study in North Carolina: Final Report . Project Number: S13/00303/OCR. Unpublished study prepared by Eurofi ns-Agroscience Services, Inc. 52p.

49173718 Lawrence, J. (2013) Dose Response Effects of MBI-20 3 DF and MBI-206 EP on the

Mealybug Destroyer (COLEOPTERA: Coccinellidae Crypt olaemus montrouzieri) in a Translational Study in North Carolina: Final Repo rt. Project Number: S13/00305/OCR. Unpublished study prepared by Eurofi ns-Agroscience Services, Inc. 53p.

49173719 Lawrence, J. (2013) Dose Response Effects of MBI-20 3 DF and MBI-206 EP on the

Predatory Spider Mite (MESOSTIGMATA: Phytoseiidae A mblyseius californicus) in a Translational Study in North Carolina: Final R eport. Project Number: S13/00484/OCR. Unpublished study prepared by Eurofi ns-Agroscience Services, Inc. 54p.

49173720 Lawrence, J. (2013) Dose Response Effects of MBI-20 3 DF and MBI-206 EP on the

Sweet Potato Whitefly Predator (COLEOPTERA: Coccine llidae Delphastus catalinae) in a Translational Study in North Caroli na: Final Report. Project Number: S13/00306/OCR. Unpublished study prepared by Eurofi ns-Agroscience Services, Inc. 53p.

49173721 Lawrence, J. (2013) Dose Response Effects of MBI-20 3 DF and MBI-206 EP on theHeat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 24 of 33

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Aphid Midge (DIPTERA: Cecidomyiidae Aphidoletes aph idimyza) in a Translational Study in North Carolina: Final Report. Project Numb er: S13/00304/OCR. Unpublished study prepared by Eurofins-Agroscience Services, Inc. 50p.

49186300 Marrone Bio Innovations (2013) Submission of Toxici ty Data in Support of the

Application for Registration of MBI-206 EP. Transmi ttal of 11 Studies.

49186301 Lawrence, J. (2013) Addendum to MRID # 491737-11 Or al Limit Test of MBI-206

EP on the Honey Bee (Hymenoptera: Apidae Apis melli fera) in a Laboratory Study in North Carolina. Project Number: S13/02866/OCR. U npublished study prepared by Eurofins-Agroscience Services, Inc. 48p.

49186302 Lawrence, J. (2013) Addendum to MRID # 491737-12 Co ntact Limit Test of MBI206 EP on the Honey Bee (Hymenoptera: Apidae Apis m ellifera) in a Laboratory Study in North Carolina: Final Report. Project Numb er: S13/02867/OCR. Unpublished study prepared by Eurofins-Agroscience Services, Inc. 42p.

49186303 Lawrence, J. (2013) Addendum to MRID # 491737-13 Do se Response Effects of

MBI-203 DF and MBI-206 EP on the Parasitic Wasp (Hy menoptera: Aphelinidae Encarsia formosa) in a Translational Study in North Carolina: Final Report. Project Number: S13/00307/OCR. Unpublished study prepared b y Eurofins-Agroscience Services, Inc. 61p.

49186304 Lawrence, J. (2013) Addendum to MRID # 491737-14 Do se Response Effects of

MBI-203 DF and MBI-206 EP on the Rove Beetle (Coleo ptera: Staphylinidae Atheta coriaria) in a Translational Study in North Carolin a. Project Number: S13/00301/OCR. Unpublished study prepared by Eurofi ns-Agroscience Services, Inc. 67p.

49186305 Lawrence, J. (2013) Addendum to MRID # 491737-15 Do se Response Effects of

MBI-203 DF and MBI-206 EP on the Bumble Bee (Hymeno ptera: Apidae Bombus sp.) in a Translational Study in North Carolina. Pr oject Number: S13/00300/OCR. Unpublished study prepared by Eurofins-Agroscience Services, Inc. 65p.

49186306 Lawrence, J. (2013) Addendum to MRID # 491737-16 Do se Response Effects of

MBI-203 DF and MBI-206 EP on the Minute Pirate Bug (Hemiptera: Anthocoridae Orius insidiosus) in a Translational Study in North Carolina: Final Report. Project Number: S13/00302/OCR. Unpublished study prepared b y Eurofins-Agroscience Services, Inc. 69p.

49186307 Lawrence, J. (2013) Addendum to MRID # 491737-17 Do se Response Effects of

MBI-203 DF and MBI-206 EP on Parasitic Wasp (Hymeno ptera: Aphelinidae Aphidius colemani) in a Translational Study in Nort h Carolina: Final Report. Project Number: S13/00303/OCR. Unpublished study prepared b y Eurofins-Agroscience Services, Inc. 59p.

49186308 Lawrence, J. (2013) Addendum to MRID # 491737-18 Do se Response Effects of

MBI-203 DF and MBI-206 EP on the Mealybug Destroyer (Coleoptera: Coccinellidae Cryptolaemus montrouzieri) in a Trans lational Study in North Carolina: Final Report. Project Number: S13/00305/O CR. Unpublished study prepared by Eurofins-Agroscience Services, Inc. 61p

.

49186309 Lawrence, J. (2013) Addendum to MRID # 491737-19 Do se Response Effects ofHeat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 25 of 33

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MBI-203 DF and MBI-206 EP on the Predatory Spider M ite (Mesostigmata: Phytoseiidae Amblyseius swirskii) in a Translationa l Study in North Carolina: Final Report. Project Number: S13/00484/OCR. Unpublished study prepared by EurofinsAgroscience Services, Inc. 66p.

49186310 Lawrence, J. (2013) Addendum to MRID # 491737-20 Do se Response Effects of

MBI-203 DF and MBI-206 EP on the Sweet Potato White fly Predator (Coleoptera: Coccinellidae Delphastus catalinae) in a Translatio nal Study in North Carolina: Final Report. Project Number: S13/00306/OCR. Unpublished study prepared by EurofinsAgroscience Services, Inc. 61p.

49186311 Lawrence, J. (2013) Addendum to MRID # 491737-21 Do se Response Effects of

MBI-203 DF and MBI-206 EP on the Aphid midge (Dipte ra: Cecidomyiidae Aphidoletes aphidimyza) in a Translational Study in North Carolina: Final Report. Project Number: S13/00304/OCR. Unpublished study pr epared by EurofinsAgroscience Services, Inc. 60p.

B. Environmental Protection Agency Risk Assessment Memoranda

U.S. EPA. 2012a. Burkholderia spp. strain A396/MBI-206 – heat inactivated cells. Memorandum from Joel V. Gagliardi, Ph.D. and John L . Kough, Ph.D. to Michael Glikes, dated July 18, 2012.

U.S. EPA. 2012b. Burkholderia spp. strain A396/MBI-206 – heat inactivated cells. Memorandum from Joel V. Gagliardi, Ph.D. and John L . Kough, Ph.D. to Michael Glikes, dated July 26, 2012.

U.S. EPA. 2013a. Heat-killed Burkholderia spp. strain A396/MBI-206. Memorandum from Joel V. Gagliardi, Ph.D. and John L. Kough, Ph.D. to Mic hael Glikes, dated March 14, 2013.

U.S. EPA. 2013b. Heat-killed Burkholderia spp. strain A396/MBI-206. Memorandum from Joel V. Gagliardi, Ph.D. and John L. Kough, Ph.D. to Mic hael Glikes, dated September 5,

2013.

U.S. EPA. 2013c. Environmental Risk Assessment for a FIFRA Section 3 Registration of Heatkilled Burkholderia spp. strain A396 cells and spent fermentation media. Mem orandum from Gail Tomimatsu, Ph.D. and Shannon Borges to Mi chael Glikes, dated December 13,

2013.

C. Other References

Centers for Disease Control and Prevention. 2009. D rinking Water - Water Treatment. Available
from http://www.cdc.gov/healthywater/drinking/public/wat er_treatment.html

.

DeFelice K, Wollenhaupt N, Buchholz D. 1993. Aquife rs and Soil Filter Effect. Available from http://extension.missouri.edu/p/WQ24

.

U.S. EPA. 2004. Primer for Municipal Wastewater Tre atment Systems. EPA 832-R-04-001.
Available from http://www.epa.gov/npdes/pubs/primer.pdf

.Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 26 of 33

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APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS

(40 CFR PART 158 – SUBPART V)

TABLE 1. Product Analysis Data Requirements for th e Active Ingredient (TGAI), Heat-killed Burkholderia sp. strain A396 cells and spent fermentation media (40 CFR § 158.2120)

Harmonized
Guideline

Number Data Requirement Results MRID No.

885.1100 Product Identity Submitted data fulfill the require ment for product identity. 482127-01
487458-01

885.1200 Manufacturing

Process Submitted data fulfill the requirement for manufact uring process. 482127-01
487458-05
487692-01
487692-03

N/A Deposition of a

Sample in a Nationally Recognized Culture
Collection Burkholderia spp. strain A396 is on deposit with the USDA Agricultural
Research Service Patent Culture Collection: NRLL B- 50319, which is not accessible to the public. N/A

885.1300 Discussion of

Formation of
Unintentional Ingredients Submitted data fulfill the requirement for analysis of samples. 482127-01
487458-05
487692-01
487692-03

885.1400 Analysis of Samples Submitted data fulfill the requirement for analysis of samples. 482127-01
487458-05
487692-01
487692-03

885.1500 Certification of

Limits Submitted data fulfill the requirement for manufact uring process. 482127-01
487458-05
487692-01
487692-03

TABLE 2. Product Analysis Data Requirements for th e End-Use Pesticide Product (EP), Venerate (40 CFR § 158.2120)

Harmonized
Guideline

Number Data Requirement Results MRID No.

885.1100 Product Identity Submitted data fulfill the requirement for product identity. Venerate contains 94.46% by weight Heat-killed Burkholderia sp. strain A396 cells and spent fermentation media. 482128-01
487685-01

488311-01
489833-05

490288-01

885.1200 Manufacturing Process Submitted data fulfill the requirement for manufact uring

process. 482128-01
487685-01

488311-01
489833-05

490288-01

N/A Deposition of a Sample in a

Nationally Recognized
Culture Collection N/A N/A

885.1300 Discussion of Formation of

Unintentional Ingredients Submitted data fulfill the requirement for analysis of samples. 482128-01
487685-01

488311-01
489833-05

490288-01Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 27 of 33

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885.1400 Analysis of Samples Submitted data fulfill the requirement for analysis of samples for purposes of FIFRA section 3(c)(5) registration. 482128-01
487685-01

488311-01
489833-05

490288-01

885.1500 Certification of Limits Limits listed on the confidential statement of form ula are adequate/acceptable. 482128-01
487685-01

488311-01
489833-05

490288-01

TABLE 3. Physical and Chemical Characteristics for the Technical Grade of the Active Ingredient (TGAI ), Heat-killed

Burkholderia spp. strain A396 cells and spent fermentation media (40 CFR § 158.2120)

Harmonized
Guideline Number Data Requirement Results MRID No.

830.6302 Color Beige

482127-01
487458-05
487692-01
487692-03 830.6303 Physical State Liquid

830.6304 Odor Mild grassy smell

830.6313 Stability to Normal and
Elevated Temperatures, Metals,

and Metal Ions Expected to be stable to elevated temperatures and to metals

and metal ions.

830.6317 Storage Stability As a term of registration, EPA is requiring additio nal information (confirmatory) on this data requirement

. 830.6319 Miscibility Non emulsifiable liquid.

830.6320 Corrosion

Characteristics As a term of registration, EPA is requiring additio nal information (confirmatory) on this data requirement

. 830.7000 pH 6.88

830.7100 Viscosity 8.48

830.7300 Density/Relative

Density/Bulk Density (Specific Gravity) 1.026 g/mL

TABLE 4. Physical and Chemical Characteristics for the End-Use Pesticide Product (EP), Venerate (40 CFR § 158.2120)

Harmonized
Guideline Number Data Requirement Results MRID No.

830.6302 Color Light brown opaque

487685-01

488311-01
489833-05 830.6303 Physical State Liquid

830.6304 Odor Fermented straw like

830.6313 Stability to Normal and

Elevated Temperatures, Metals, and Metal Ions Unstable to elevated temperatures and to metals and
metal ions.

830.6317 Storage Stability As a term of registration, EPA is requiring additio nal information (confirmatory) on this data requirement

. 830.6319 Miscibility Miscible with water

830.6320 Corrosion Characteristics As a term of registration, EPA is requiring additio nal information (confirmatory) on this data requirement

. 830.7000 pH 6.61 830.7100 Viscosity 12.0

830.7300 Density/Relative

Density/Bulk Density (Specific Gravity) 1.015 g/mLHeat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 28 of 33

Biopesticides Registration Action Document

TABLE 5. Toxicology Data Requirements for the Acti ve Ingredient Heat-killed Burkholderia spp. strain A396 cells and spent

fermentation media and the End Use Pesticide Produc t Venerate (40 CFR § 158.2140)

Harmonized
Guideline

Number Data Requirement Results MRID No.

Tier I

885.3050 Acute Oral

Toxicity/Pathogenicity Waived due to heat killing of the active ingredient after fermentation and batch verification by enrichment that no live Burkholderia spp. strain A396 cells are present in any products. Classification: Acceptable
TOXICITY CATEGORY IV 482127-08
482127-09

488310-02
489833-02

885.3150 Acute Pulmonary

Toxicity/Pathogenicity Waived due to heat killing of the active ingredient after fermentation and batch verification by enrichment that no live Burkholderia spp. strain A396 cells are present in any products. Classification: Acceptable
TOXICITY CATEGORY IV 482127-08
482127-09
488310-02
489833-02

885.3200 Acute Injection

Toxicity/ Pathogenicity Waived due to heat killing of the active ingredient after fermentation and batch verification by enrichment that no live Burkholderia spp. strain A396 cells are present in any products. Classification: Acceptable
TOXICITY CATEGORY IV 482127-08
482127-09

488310-02
489833-02

885.3400 Hypersensitivity

Incidents The applicant reported that no hypersensitivity inc idents, including

immediate-type or delayed-type reactions of humans and domestic animals, occurred during research, development, or testing o f the TGAI. Any future hypersensitivity incidents must be reported to EPA (refer to test note #3 of 40 CFR § 158.2140(d)).

885.3500 Cell Culture Not required because Heat-killed Burkholderia sp. strain A396 cells and spent fermentation media is not a virus (refer to t est note #4 of 40 CFR § 158.2140(d)). N/A

870.1100 Acute Oral Toxicity Oral LD 50 female rats > 5,000 mg/kg body weight (bw) Classification: Acceptable
TOXICITY CATEGORY IV 491737-01

870.1200 Acute Derm al Toxicity Dermal LD 50 combined (female rats) > 5,050 mg/kg bw Classification: Acceptable
TOXICITY CATEGORY IV 491737-02

870.1300 Acute Inhalation

Toxicity LC 50 combined (male and female rats) >5.22 mg/L

Classification: Acceptable
TOXICITY CATEGORY IV 491737-03

870.2400 Acute Eye Irritation Venerate was non-irritating to the eyes of rabbits. Classification: Acceptable
TOXICITY CATEGORY IV 491737-04

870.2500 Primary Dermal

Irritation The EP test substance was slightly irritating to th e skin of guinea pigs during

the first hour of exposure. Classification: Acceptable
TOXICITY CATEGORY IV 491737-05

870.2600 Skin Sensitizaiton Venerate was non-irritating to t he skin of guinea pigs. 491737-06

Tiers II and III Not required for Heat-killed Burkholderia spp. strain A396 cells and spent fermentation media , based on the lack of acute toxicity/pathogenicity in the Tier I studies.Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 29 of 33

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TABLE 6. Nontarget Organism Toxicity and Environme ntal Fate Data Requirements for the Technical Grade of the Active Ingredient (TGAI), Heat-killed Burkholderia spp. strain A396 cells and spent fermentation media (40 CFR § 158.2150)

Harmonized Guideline

Number Data Requirement Results MRID No.

Tier I

885.4050 Avian Oral Toxicity The resubmitted information and data did not support equivalence of the test material used in these studies to the test substances per the amended manufacturing processes. The 29-day

LD 50 (conducted on 23-day old chicks) was determined > 112.5 mg a.i./kg bw. Classification: Supplemental 48212711

48831001
49173700

LD 50 >2,000 mg a.i./kg bw in a 14-day limit study (conducted on 133-day old adults).
Classification: Supplemental/Not Upgradeable 49173707

850.2100 Avian Oral Toxicity 14-day oral study on bobwhite quail; no mortalities or other biological endpoints

noted. LD 50 >2,000 mg a.i./kg bw; Classification: Acceptable 49173707

885.4100 Avian Inhalation Toxicity/Pathogenicity Not required as the nature of the microbial pesticide does not indicate potential pathogenicity to birds or relatedness to any known bird pathogens (refer to test note #3 of 40 CFR § 158.2150(e)). 49173702

885.4150 Wild Mammal Toxicity/Pathogenicity Tests required by 40 CFR § 158.2140 are adequate and appropriate for assessment of
hazards to wild mammals. Studies submitted with laboratory rats indicate no adverse effects due to oral exposure. Classification: Acceptable for wild mammal
risk assessment 49173701
49173702
49173703

885.4200 Freshwater Fish Toxicity The resubmitted information and data did not support equivalence of the test material used in these studies to the test substances per the amended manufacturing process. A 30-day LC 50 was determined > 81 mg a.i./L (fathead minnow). Classification: Supplemental 48212712
48625801

48831001
49173700

The resubmitted information and data did not support equivalence of the test material used in these studies to the test substances per the amended manufacturing process. No rainbow trout mortalities reported at the single-limit dose tested (96-hr LC 50 > 100 mg MBI-206EP/L). Classification: Supplemental 48625801

48831001
49173700

NOEC and EC 50 > 1,000 mg a.i. after 96-hr under static conditions (rainbow trout). Classification: Acceptable 49173709

885.4240 Freshwater Invertebrate Toxicity The re-submitted information and data did not

support equivalence of the test material used in these studies to the test substances per the amended manufacturing process. The 21-day

LC 50 was determined to be > 81 mg a.i. /L. Classification: Supplemental 48212713

48831001
48745803

49173700Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 30 of 33

Biopesticides Registration Action Document

TABLE 6. Nontarget Organism Toxicity and Environme ntal Fate Data Requirements for the Technical Grade of the Active Ingredient (TGAI), Heat-killed Burkholderia spp. strain A396 cells and spent fermentation media (40 CFR § 158.2150)

Harmonized Guideline

Number Data Requirement Results MRID No.

850.1010 Freshwater Invertebrate Toxicity 48-hr acute toxicity study conducted under static

conditions; 48 hr EC 50 = 48.17 mg a.i./L. Classification: Acceptable 49173708

885.4280 Estuarine/Marine Fish and Invertebrate Testing Not required as the microbial pesticide will not be applied directly to water and is not expected to
enter marine/estuarine environments in amounts that would result in significant concentrations (refer to test note #6 of 40 CFR § 158.2150(e)). N/A

885.4300 Nontarget Plant Testing This study is not quantitative and is of insufficient duration and sample size to assess potential risks to nontarget plants outside of intended “agricultural environments.” Testing was not conducted at
maximum hazard concentrations. Phytotoxicity symptoms ranged from none to mild for tested monocots; and from mild to
moderate for tested dicots at two concentrations. Severe phytotoxicity was noted for English peas at the 1.5X lowest
labeled concentration of MBI-206 EP. Classification: Invalid/Incomplete 48212802
48745803
48745805

48831001

885.4150 Nontarget Plant Testing The study is inconclusive because definitive endpoints were not observed on tested plants in the study and the risk to nontarget plant species cannot be determined. Phytotoxic damage on tested species ranged from cuticle damage on tomato to brown spotting on soybean, radish, cabbage, lettuce, cucumber, and corn. No phytotoxic symptoms were observed on oat, ryegrass, and onion. Classification: Inconclusive/Incomplete 49173710

885.4340 Nontarget Insect Testing: Green Lacewing This test was not conducted at maximum hazard concentrations to obtain biologically significant endpoints. Classification: Invalid/Incomplete 48212714
48831001

48212802

885.4340 Nontarget Insect Testing: Parasitic Wasp The test results are inconclusive for the 4-day

Study; the Agency cannot determine the toxicity of the insecticidal endotoxin(s) to parasitic wasps. Classification: Invalid/Incomplete 48745804

48831001

48212802

885.4340 Nontarget Insect Testing: Ladybird Beetle The test results are inconclusive in this 7-day

study; the Agency cannot determine the toxicity of the insecticidal endotoxin(s) to ladybird beetles. Classification: Invalid/Incomplete 48212716
48745803

48831001Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 31 of 33

Biopesticides Registration Action Document

TABLE 6. Nontarget Organism Toxicity and Environme ntal Fate Data Requirements for the Technical Grade of the Active Ingredient (TGAI), Heat-killed Burkholderia spp. strain A396 cells and spent fermentation media (40 CFR § 158.2150)

Harmonized Guideline

Number Data Requirement Results MRID No.

885.4340 Nontarget Insect Testing: Parasitic Wasp The toxicity to parasitic wasp cannot be

defined from these results with certainty
because of several limitations of the experimental approach and design that present uncertainty in ensuring that the results observed are due to the toxicity of the
test material. Classification: Unacceptable 49173713

885.4340 Nontarget Insect Testing:
Rove Beetle The toxicity to rove beetle cannot be defined

from these results with certainty because of several limitations of the experimental approach and design that present uncertainty in ensuring that the results observed are due to the toxicity of the test material. Classification: Unacceptable 49173714

885.4340 Nontarget Insect Testing:

Bumblebee The toxicity to rove beetle cannot be defined

from these results with certainty because of several limitations of the experimental approach and design that present uncertainty in ensuring that the results observed are due to the toxicity of the test material Classification: Unacceptable 49173715

885.4340 Nontarget Insect Testing: Minute Pirate Bug The toxicity to rove beetle cannot be defined

from these results with certainty because of several limitations of the experimental approach and design that present uncertainty in ensuring that the results observed are due to the toxicity of the test material. Classification: Unacceptable 49173716

885.4340 Nontarget Insect Testing: Parasitic Wasp The toxicity to parasitic wasp cannot be

defined from these results with certainty
because of several limitations of the experimental approach and design that present uncertainty in ensuring that the results observed are due to the toxicity of the
test material. Classificaion: Unacceptable 49173717

885.4340 Nontarget Insect Testing:

Adult Melybug Destroyer The toxicity to mealybug destroyer cannot be defined from these results with certainty
because of several limitations of the experimental approach and design that present uncertainty in ensuring that the results observed are due to the toxicity of the
test material. Classificaion: Unacceptable 49173718Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 32 of 33

Biopesticides Registration Action Document

TABLE 6. Nontarget Organism Toxicity and Environme ntal Fate Data Requirements for the Technical Grade of the Active Ingredient (TGAI), Heat-killed Burkholderia spp. strain A396 cells and spent fermentation media (40 CFR § 158.2150)

Harmonized Guideline

Number Data Requirement Results MRID No.

885.4340 Nontarget Insect Testing: Predatory Spider Mite The toxicity to predatory spider mite cannot be defined from these results with certainty
because of several limitations of the experimental approach and design that present uncertainty in ensuring that the results observed are due to the toxicity of the
test material. Classification: Unacceptable 49173719

885.4340 Nontarget Insect Testing: Sweet Potato Whitefly

Predator The toxicity to sweet potato whitefly cannot

be defined from these results with certainty
because of several limitations of the experimental approach and design that present uncertainty in ensuring that the results observed are due to the toxicity of the
test material. Classification: Unacceptable 49173720

885.4340 Nontarget Insect Testing: Aphid Midge The toxicity to aphid midge cannot be

defined from these results with certainty
because of several limitations of the experimental approach and design that present uncertainty in ensuring that the results observed are due to the toxicity of the
test material. Classification: Unacceptable 49173721

885.4380 Honey Bee Testing Bee mortalities were reported for all groups, including the negative control. Therefore, the study results are considered inconclusive and the honeybee data requirement is incomplete. Classification: Invalid/Incomplete 48212717
48745803

48831001

885.3020 Honey Bee Testing The toxicity to honeybees cannot be defined from these results with certainty because of limitations of the experimental approach and design that present uncertainty in ensuring that the results observed are due to the toxicity of the test
material. Classification: Unacceptable 49173712
49186302

885.3020 Honey Bee Testing The toxicity of MBI-206 to honeybees cannot be defined from these results with certainty because of limitations of the experimental approach and design that present uncertainty in ensuring that the results observed are due to the toxicity of the
MBI-206 test material. Classification: Unacceptable 49173711
49186301

885.3020 Honey Bee Testing The review is based on an unsigned study report and has “Draft Version” on the cover page. The study may be upgraded following review of a Final Study Report and applicant’s response to questions. Bee mortalities were reported. Classification: Supplemental; may be upgraded. N.A.

Tiers II, III, and IV Not required for Heat-killed Burkholderia spp. strain A396 cells and spent fermentation media based on the acceptability of the data and other information provided for Tier I.Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 33 of 33

Biopesticides Registration Action Document

APPENDIX B. PESTICIDE PRODUCT

EPA File Symbol Registration

Name Percentage

Active Ingredient Formulation

Type Use Site(s) Method(s) of Application Application Rate(s) Target Pest

84059-RG MBI-206

TGAI 100% Manufacturinguse product manufacturing NA NA NA

84059-RU Venerate 94.46% End-use product Agricultural

Seed treatment
Turf/Ornamental
Residential Foliar spray

Aerial Chemigation
Tank mix
Soil treatment

Seed treatment Hose-end sprayer 2-8 quarts per acre

Soil treatment: 1-2 gallons per acre

Seed treatment: 8 ounces per 100 pounds
of seed

Home and Garden: 2 tablespoons per gallon
of water Insect pests and
nematodes