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, 2014 

 
Heat-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 Ingredient, 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 ................................................................................................................ 33 
Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 3 of 33

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, Human 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 Branch 

Michael Glikes Regulatory Action Leader 
Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 4 of 33

Biopesticides Registration Action Document 

I. EXECUTIVE SUMMARY 

Background

In August 2010, Marrone Bio Innovations, Inc., submitted 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 United States Environmental Protection 

Agency (EPA) under section 3 of the Federal Insecticide, Fungicide, and Rodenticide Act 

(FIFRA). Concurrently with this application, MBI filed 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 these 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 insecticide. As an 

insecticide, it controls many foliar feeding pests, such as coleopteran pests: soft-bodied insect 

pests, such as aphids, mites and whiteflies; and nematodes. The proposed manufacturing product 

(MP) containing this new TGAI is called “MBI-206 TGAI.” 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 biological insecticide primarily applied as a foliar spray. 

The mode of action for this killed microbial pesticide is explained by bioactive metabolites that 

have insecticidal activity. The proposed EP is intended 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 and ornamental bedding plants and flowers through ground, chemigation and 

aerial equipment. Intended uses also include seed treatments with the product before planting, as 

well as homes and gardens.

EPA scientists reviewed product analysis, toxicology, 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 that, 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 under FIFRA section 3(c)(5). Terms of the 

proposed registrations would require the manufacturer within one year after registration to 

submit additional data on the EP’s storage stability, 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) registration, the product analysis data requirements 

for heat-killed Burkholderia spp. strain A396 cells and spent fermentation media, including 

product chemistry and composition, analysis of samples, physical and chemical characteristics, 

and additional data concerning the manufacturing process 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

Biopesticides Registration Action Document 

Toxicology

The applicant submitted adequate mammalian toxicology data and information to support the 

pesticide product. Acute toxicity studies showed that, 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 incidents 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 absence 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 concluded 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. Data 

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-killed 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-killed 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 pesticide product, Venerate (foliar spray and/or 

seed directed; no aquatic applications); (2) the filtering effect of many particulate soil types; and 

(3) the conditions (e.g., filtration and pH adjustments) water is subjected to in wastewater 

treatment systems and drinking water facilities. Based 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 unreasonable adverse effects. 

Occupational Exposure

Despite the low toxicological profile of heat-killed 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, may be subject to prolonged or repeated 

exposure to the active ingredient. The proposed product label directs handlers working with heatkilled Burkholderia spp. strain A396 cells and spent fermentation media in agricultural settings 

to wear a long-sleeved shirt, long pants, waterproof gloves, socks, shoes and a dust/mist filtering 

respirator meeting National Institute for Occupational Safety and Health (NIOSH) standards of at 

least N-95, R-95, or P-95. For future products, EPA may require additional PPE such as 

protective eyewear on a product-specific basis. 

Nontarget Organisms

The applicant submitted studies of nontarget organism 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 determined that adverse effects to most nontarget 

organisms are not anticipated from the proposed pesticidal uses of heat-killed Burkholderia spp. 
Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 6 of 33

Biopesticides Registration Action Document 

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 because the studies the applicant submitted on 

honeybees and 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. EPA 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 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 applications: new active ingredients; first food 

uses; first outdoor uses; first residential uses; or any other registration actions for which the 

Agency believes there may be significant public interest. 

 

Consistent with the policy of making registration actions more transparent, the pesticide product 

containing heat-killed Burkholderia spp. strain A396 cells and spent fermentation media, a new 

active ingredient, is subject to a 15-day comment period. In addition to containing a new active 

ingredient, the registration of the pesticide product 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 registration actions, and accessed through either 

http://www.regulations.gov or http://www.epa.gov/pesticides/regulating/registration-status.html, 

is EPA-HQ-OPP-2011-0010. The following documents are 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 (BRAD); 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 File 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 appropriate to issue the proposed registrations. The 

basis for proposed decisions can be found in the risk assessment for heat-killed Burkholderia 

spp. strain A396 cells and spent fermentation media, which 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: 006534 
Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 7 of 33

Biopesticides Registration Action Document 

 Type of Pesticide: Microbial Pesticide –Insecticide. 

 See Appendix B for specific information (e.g., 

use sites, application rates, methods of application, 

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. (address: 2121 Second Street, Suite B-107; 

Davis, California 95618) submitted an application to register a manufacturing product, MBI-206 

TGAI (EPA File Symbol 84059-RG) and an end-use pesticide product, Venerate (EPA File 

Symbol 84059-RU), under FIFRA section 3. On February 2, 2011 (76 FR 5805), the EPA 

announced receipt of the applications to register pesticide products containing a new active 

ingredient, one for manufacturing into end-use products, 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 and ornamental bedding plants and flowers 

through ground, chemigation and aerial equipment. Intended uses also include seed treatments 

with the product before planting, as well as home and 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 under Federal Food, Drug, and Cosmetic Act 

(FFDCA) section 408(d), Marrone Bio Innovations, Inc., 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, 2011 (76 FR 10584), the EPA announced that 

Marrone Bio Innovations, Inc., proposed an exemption 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 microbial 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 indication of the usefulness of the information 

contained in the documents for risk assessment. A rating of “acceptable” indicates the study is 

scientifically sound and is useful for risk assessment. A “supplemental” rating indicates the data 

provide some information that can be useful for risk assessment. The studies may have certain 
Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 8 of 33

Biopesticides Registration Action Document 

aspects determined not to be scientifically acceptable (“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 “acceptable.” If there is simply a “supplemental” 

rating, the reviewer will often state that the study 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 that 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 information submitted to the EPA in support of a 

pesticide registration. The EPA classifies the active 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 applicant 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 registration. 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 manufacturing-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) and 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/Pathogenicity – Rat (Harmonized Guideline 

885.3150; and Acute Injection Toxicity/Pathogenicity (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 verification by enrichment; hence, no live cells are 

present in any products. 

Hypersensitivity Incidents (Harmonized Guideline 885.3400): The applicant reported that no 

hypersensitivity incidents, including immediate-type or delayed-type reactions of humans and 

domestic animals, occurred during research, development, or testing of heat-killed Burkholderia 
Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 9 of 33

Biopesticides Registration Action Document 

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 not 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-Dawley, 

albino rats a single dose of Venerate (94.46 %, undiluted 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 deaths 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 when dosed at approximately 5000 mg/kg bw. 

The EPA classified the acute oral toxicity/pathogenicity as Toxicity Category IV, the lowest 

toxicity category, and the Agency rated the study as acceptable. 

Acute Dermal Toxicity - Rat (Harmonized Guideline 870.1200; 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 14 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 EPA classified the dermal toxicity as Toxicity 

Category IV, and the Agency rated the study as acceptable. 

Acute Inhalation Toxicity– Rat (Harmonized Guideline 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 articles for 4 hours at concentrations of 5.22 mg/L. 

None of the rats died after 14 days, and the researchers 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 as acceptable. 

Acute Eye Irritation Study - Rabbit (Harmonized Guideline 870.2400; MRID No 491737-04): 

Researchers instilled 0.1ml of Venerate into the conjunctival sac of the right eye of 2 male and 2 

female, 10 week-old albino rabbits for 24 hours. They 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 irritation 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 trunks of 2 male and 1 female, 10 week-old 

albino rabbits and then covered the exposed skin for 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 hour observations. The EPA classified the 

primary dermal irritation as Toxicity Category IV and rated the study as acceptable. 

 
Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 10 of 33

Biopesticides Registration Action Document 

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

ReproductiveFertility Effects, Carcinogenicity, Immunotoxicity, 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 designate.” FFDCA section 408(p)(4) authorizes the 

Administrator, by order, to exempt from the requirements 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 similar to an effect produced by a naturally 

occurring estrogenic substance. Between October 2009 and February 2010, EPA issued test 

orders/data call-ins for the first group of 67 chemicals, 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 ingredients on the initial list to 

be screened under the EDSP. 

The EPA believes that heat-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 heat-killed Burkholderia sp. strain A396 cells 

and spent fermentation media from the requirements of the Section 408(p)(4) EDSP. In the event 

the EPA does determine to exempt this substance from the EDSP, an order will be issued. For 

further information on the status of the EDSP, the policies and procedures, the list of 67 

chemicals, the test guidelines and the Tier 1 screening battery, please visit our website: 

http://www.epa.gov/endo/. 

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

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 certainty that no harm will result from aggregate 

exposure to the pesticide chemical residue, including all anticipated dietary exposures and all 

other exposures for which there is reliable information.” This includes exposure through drinking 

water and in residential settings, but does not include occupational exposure. Pursuant to section 

408(c)(2)(B) of FFDCA, in establishing or maintaining 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 give special consideration to exposure of 

infants and children to the pesticide chemical residue in establishing a tolerance exemption, and 

to “ensure that there is a reasonable certainty that no harm will result to infants and children from 
Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 11 of 33

Biopesticides Registration Action Document 

aggregate exposure to the pesticide chemical residue....” Additionally, section 408(b)(2)(D) of 

FFDCA requires that 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 performs a number of analyses to determine the risks from aggregate exposure to 

pesticide residues. First, the Agency determines the toxicity of a pesticide. Second, the EPA 

examines exposure to the pesticide through food, drinking water, and through other exposures 

that occur as a result of pesticide use in residential 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 sensitivities 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 for heatkilled Burkholderia spp. strain A396 cells and spent fermentation media have been fulfilled. 

 

 a. Aggregate Exposure 

In examining aggregate exposure, section 408 of FFDCA 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-killed

Burkholderia spp. strain A396 cells and spent fermentation media, a naturally occurring soil 

bacterium, is anticipated to be negligible. Due to the ubiquitous 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 humans 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 are foliar, thereby limiting contact with surface 

water by drift and runoff. Furthermore, ground water is not expected to have significant exposure 

to heat-killed Burkholderia spp. strain A396 cells and spent fermentation media since, like other 

microorganisms, this microbial pesticide would likely be filtered out by the particulate nature of 

many soil types. If it were to be transferred to surface or ground waters that are intended for 

eventual human consumption (e.g., through spray drift 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, including 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-killed Burkholderia spp. strain A396 cells and 

spent fermentation media is present in drinking water (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 Burkholderia 
Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 12 of 33

Biopesticides Registration Action Document 

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 occurrence in the environment, the EPA 

determined that non-occupational exposure to the bacterium likely already is occurring. 

Additional exposure to the microorganism, due to pesticidal 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 not 

toxic (acute dermal toxicity and acute pulmonary toxicity/pathogenicity), non-irritating (primary 

dermal irritation), and not pathogenic (acute pulmonary 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 Common Mechanism of Toxicity 

Section 408(b)(2)(D)(v) of FFDCA requires that, when 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 purposes 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 other substances. Thus, section 

408(b)(2)(D)(v) of the FFDCA does not apply. For information regarding the EPA’s efforts to 

determine which chemicals have a common mechanism of 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 the EPA shall assess the available 

information about consumption patterns among infants 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 shall apply an additional tenfold (10X) 

margin of exposure (safety) for infants and children in the case of threshold effects to account for 

prenatal and postnatal toxicity and the completeness of the database on toxicity and exposure 

unless the EPA determines that a different margin of exposure (safety) will be safe for infants 

and children. This additional margin of exposure (safety) is commonly referred to as the Food 

Quality Protection Act Safety Factor. In applying this 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

Biopesticides Registration Action Document 

(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 labeled in accordance with good agricultural 

practices. As a result, the EPA concludes that no additional 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 EPA analysis as presented directly above, the 

Agency is able to conclude that there is a reasonable certainty that no harm will result to the 

United States population, including infants and children, 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 agricultural 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 conclusion 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 pathogenic potential to mammals, including 

infants and children. 

3. Occupational Exposure and Risk Characterization

The EPA does not expect handler exposure to heat-killed Burkholderia spp. strain A396 cells 

and spent fermentation media to pose any undue risk. Regardless, appropriate personal protective 

equipment (PPE) and precautionary statements are required 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 filtering respirator meeting National Institute for 

Occupational Safety and Health (NIOSH) standards of at least N-95, R-95, or P-95. For future 

products containing this microorganism as an active ingredient, the EPA may require additional 

PPE such as protective eyewear on a product-specific 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 for 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 used in 

accordance with EPA-approved labeling. 

C. Environmental Assessment (40 CFR § 158.2150) 

Data and other information (e.g., scientific literature) submitted by the applicant to support 

requests to waive nontarget organism testing for heat-killed Burkholderia spp. strain A396 cells 

and spent fermentation media are sufficient to fulfill the relevant microbial pesticide data 

requirements and for risk assessment purposes. Further testing of nontarget organisms at higher 

tier levels (i.e., Tiers II, III, and IV) is not required. 
Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 14 of 33

Biopesticides Registration Action Document 

The EPA performed an environmental risk assessment based on data and other information 

provided by the applicant, and the Agency determined that adverse effects to nontarget 

organisms are not anticipated from the pesticidal uses of heat-killed Burkholderia spp. strain 

A396 cells and spent fermentation media. For honeybees and nontarget insects, however, the 

Agency has insufficient information to conclude that 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 registration, 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 effects to nontarget insects and plants, EPA has 

concluded that additional analyses would be necessary to make a determination for these species. 

For a summary of the generic data requirements described 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 containing heat-killed Burkholderia spp. strain A396 

cells and spent fermentation media include foliar sprays and soil directed products, birds and 

wild mammals could be exposed. Such exposure is expected to occur through ingestion of food 

items that contact the active ingredient, or incidental ingestion of soil or drinking water (e.g., 

puddles) that contain pesticide residues, or contact with treated surfaces. Birds and wild 

mammals were not adversely affected when exposed at high dosage rates, as demonstrated in 

bird studies (MRID 49173707), and adverse effects were 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 proposed end product’s label. 

 Nontarget Insects (Harmonized Guideline 885.4340), Honey Bees (Harmonized Guideline 

885.4380), and Plants (Harmonized Guideline 885.4300): 

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 items. The EPA classified studies submitted on 

nontarget insects as either Invalid/Incomplete or Unacceptable, honey bees studies as 

Unacceptable, and nontarget plants as Invalid/Incomplete (see Table 6 on pages 29-33 for 

MRIDs). These classifications resulted from limitations in the studies’ approaches and 

experimental designs. For example, the researchers observed insects for only 24 hours after 

exposure to the pesticide while insecticidal activity could reasonably be expected after 24 hours. 
Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 15 of 33

Biopesticides Registration Action Document 

Because these studies did not clearly demonstrate hazard to these nontarget organisms, the risks 

posed cannot be determined without uncertainty. Therefore, the Agency is requiring 

precautionary and mitigating text on the proposed end-use product’s label to protect pollinating 

insects and beneficial insects used in IPM practices 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 phytotoxicity (e.g. no plant recovery after leaf burn, 

chlorosis or leaf curling) or reduced seed emergence 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, Freshwater Invertebrates (Harmonized 

Guideline 885.1010), Estuarine/Marine Fish and Invertebrates (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 proposed use on cranberries or from other 

applications that runoff and drift into aquatic areas. Applications from foliar sprays, in particular,

could result in some runoff and drift of the applied heat-killed Burkholderia spp. strain A396 

cells and spent fermentation media into aquatic areas, but the concentrations resulting from drift 

and runoff are expected to be a small fraction of the amount applied. Studies conducted on 

rainbow trout and freshwater invertebrates exposed to the proposed end-use product were 

classified as Acceptable by EPA scientists. Therefore, adverse effects to freshwater fish and 

invertebrates are not expected from proposed applications 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, further 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 exposed to the active ingredient, heat-killed 

Burkholderia spp. strain A396 and spent fermentation media in the proposed 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 listed endangered and threatened (“listed”) 

mammals, birds, and freshwater and marine/estuarine animals and for their designated critical 

habitats. Because of uncertainties and data gaps associated 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 analysis 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

Biopesticides Registration Action Document 

V. ENVIRONMENTAL JUSTICE 

The EPA seeks to achieve environmental justice—the fair treatment and meaningful involvement 

of all people regardless of race, color, national origin, or income—with respect to the 

development, implementation, and enforcement of environmental laws, regulations, and policies. 

Fair treatment means that no group of people, including racial, ethnic, or socioeconomic groups, 

should bear a disproportionate share of the negative environmental consequences resulting from 

industrial, municipal, and commercial operations or the execution of federal, state, local, and 

tribal environmental programs and policies. Meaningful involvement means that (1) potentially 

affected community residents have an appropriate opportunity to participate in decisions about a 

proposed activity that will affect their environment and/or health; (2) the public’s contribution 

can influence the regulatory agency’s decision; (3) the concerns of all participants involved will 

be considered in the decision-making process; and (4) the decision-makers seek out and facilitate 

the involvement of those potentially affected. The EPA has this goal for all communities and 

persons across the United States. 

To help address potential environmental justice issues, the EPA sought information on any 

groups or segments of the population who, as a result 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 registration of a pesticide provided that all the 

following determinations are made: 

 (1) Its composition is such as to warrant the proposed claims for it; 

 (2) Its labeling and other material required to be submitted comply with the 

 requirements of FIFRA; 

 (3) It will perform its intended function without unreasonable adverse effects on 

 the environment; AND 

(4) When used in accordance with widespread and commonly recognized 

 practice, it will not generally cause unreasonable adverse effects on the 

 environment. 

To satisfy criterion 1, the proposed products containing 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 unreasonable adverse effects on the environment 

when used according to label instructions. Criterion 2 is satisfied by the proposed product labels, 

as well as the data and information presented in this document. The Agency believes that the 

proposed heat-killed Burkholderia spp. strain A396 cells and spent fermentation media pesticide 

products will not cause any unreasonable adverse effects on the environment. In fact, Venerate is 

likely to provide protection against many insect pests, 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 unreasonable adverse effects when used according 
Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 17 of 33

Biopesticides Registration Action Document 

to label instructions. Therefore, the proposed products, 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 registration 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 pesticide products containing heat-killed

Burkholderia spp. strain A396 cells and spent fermentation media –may be released for 

shipment, the registrant will be required to provide 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 information on the products within six months of 

registration. 

Additionally, as terms of the proposed Venerate registration, Marrone Bio Innovations, Inc., will 

be required to submit studies on: 

• Storage Stability (OCSPP 830.6317) and Corrosion Characteristics 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 Hypersensitivity Incidents 

Notwithstanding the information stated in the previous 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. Reports of all incidents of adverse effects to the 

environment must be submitted to the EPA under the provisions stated in FIFRA section 6(a)(2). 

Additionally, all incidents of hypersensitivity (including both suspected and confirmed incidents) 

must be reported to 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

Biopesticides Registration Action Document 

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 Rodenticide Act 

FR Federal Register 

g/mL grams per milliliter 

lb/gal pounds per gallon 

LD50 median lethal dose. A statistically derived single dose that can be expected 

 to cause death in 50% of the test animals when administered by the route 

 indicated (oral, dermal, or inhalation). It is expressed as a weight of 

 substance per unit weight of animal (e.g., mg/kg).

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 and 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 States
Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 19 of 33

Biopesticides Registration Action Document 

IX. BIBLIOGRAPHY 

A. Studies Submitted to Support the Active Ingredient, 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 Product Chemistry, Toxicity, and 

Exposure and Risk Data in Support of the Application 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 TGAI. 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 Stillmeadow, Inc. 11 p. 

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

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

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

Project Number: 13038/09. Unpublished study prepared 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 prepared by Stillmeadow, Inc. 16 p. 

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

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

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

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

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

Study (in Rats): Final Report. Project Number: 13055/09. 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. Unpublished 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 prepared by Stillmeadow, Inc. 10 p. 

48212711 Younger, C. (2009) MOI 010: Acute Oral Toxicity Study 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. Project Number: 13286/09. 

Unpublished study prepared by Stillmeadow, Inc. 19 p. 

48212713 Hartwell, T. (2009) Microbial Pest Control Agent (MPCA) 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

Biopesticides Registration Action Document 

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

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

18 p. 

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

Insect Microbial Testing: Final Report. Project Number: 13903/10. Unpublished 

study prepared by Stillmeadow, Inc. 14 p. 

48212716 Younger, C. (2009) MOI 010: Ladybird Beetle Non-Target 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 mellifera, Non-Target Insect 

Microbial Testing: Final Report. Project Number: 13905/10. Unpublished study 

prepared by Stillmeadow, Inc. 12 p. 

48212718 Pitts, K. (2010) Response to Tier 1 Microbial Pesticide 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 Product Chemistry, Fate, and 

Toxicity Data in Support of the Application for Registration of MBI-206 EP. 

Transmittal of 3 Studies. 

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

prepared by Marrone Bio Innovations. 182 p. 

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

Unpublished study prepared by Marrone Bio Innovations. 14 p. 

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

48625801 Brunner, S. (2011) MBI-206 EP: Rainbow Trout (Oncorhynchus 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 Microbial Identification Study of 

MRID 48212701. Unpublished study prepared by Marrone Bio Innovations. 450p. 

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

Genus Burkholderia with Relevance to Agricultural and 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 Innovations. 44p. 

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

Microbial Testing: Final Report. Project Number: 13903/10. Unpublished study 

prepared by Stillmeadow, Inc. 14p. 

48745805 Pitts, K. (2012) MBI-206 TGAI Response to Completeness 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

Biopesticides Registration Action Document 

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

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

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

Clarifications/Deficiencies. Unpublished study prepared by Marrone Bio Innovations. 

13p. 

48768501 Lam, R. (2012) MBI-206 EP: Amended Product Chemistry. 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 Innovations. 58p. 

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

Innovations. 21p. 

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

(Burkholderia sp. Strain A396). Project Number: 8202/162. Unpublished study 

prepared by Marrone Bio Innovations. 25p. 

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

Unpublished study prepared by Marrone Bio Innovations. 362p. 

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

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

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

Characteristics Study. Unpublished study prepared by Marrone Bio Innovations. 73p. 

48983300 Marrone Bio Innovations (2012) Submission of Product 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 Response: Microbial Identification 

Addendum to MRID Numbers 482127-01 and 487458-01. Unpublished study 

prepared by Marrone Bio Innovations. 253p. 

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

Unpublished study prepared by Stillmeadow, Inc. 13p. 

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

of Burkholderia sp. Strain A396. Project Number: 4207. Unpublished study prepared 

by Marrone Bio Innovations. 22p. 

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

MBI/MB/2001/SOP, MBI/FM/2001/SOP, MBI/M/4002/SOP. Unpublished 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 prepared by Marrone Bio 

Innovations. 85p. 

49028800 Marrone Bio Innovations (2012) Submission of Product 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

Biopesticides Registration Action Document 

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

2012. Unpublished study prepared by Marrone Bio Innovations. 8p. 

49028900 Marrone Bio Innovations (2012) Submission of Product 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 Manufacturing Process Study-December 

2012. Unpublished study prepared by Marrone Bio Innovations. 19p. 

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

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

49171901 Bowen, R. (2013) MBI-206 Sterile Cell Lysate: Reverse Mutation in Five Histidinerequiring Strains of Salmonella typhimurium Using a Treat and Plate Methodology. 

Project Number: 8278780/OCR. Unpublished study prepared by Covance 

Laboratories, Ltd. 37p. 

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

Bobwhite Quail. Project Number: 17296/13/OCR. Unpublished study prepared by 

Stillmeadow, Inc. 16p. 

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

Project Number: 17311/13/OCR. Unpublished study prepared by Stillmeadow, Inc. 

14p. 

49173709 Mikulas, J. (2013) MBI-206 EP: Rainbow Trout (Oncorhynchus 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 Toxicity, 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 on 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

Biopesticides Registration Action Document 

Final Report. Project Number: S13/02866/OCR. Unpublished 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 Laboratory Study in North Carolina: 

Final Report. Project Number: S13/02867/OCR. Unpublished 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 formosa) in a Translational 

Study in North Carolina: Final Report. Project Number: S13/00307/OCR. 

Unpublished study prepared by Eurofins-Agroscience Services, Inc. 52p. 

49173714 Lawrence, J. (2013) Dose Response Effects of MBI-203 DF and MBI-206 EP on the 

Rove beetle (COLEOPTERA: Staphylinidae Atheta coriaria) in a Translational Study 

in North Carolina: Final Report. Project Number: S13/00301/OCR. Unpublished 

study prepared by Eurofins-Agroscience Services, Inc. 58p. 

49173715 Lawrence, J. (2013) Dose Response Effects of MBI-203 DF and MBI-206 EP on the 

Bumble Bee (HYMENOPTERA: Apidae Bombus sp.) in a Translational Study in 

North Carolina: Final Report. Project Number: S13/00300/OCR. Unpublished study 

prepared by Eurofins-Agroscience Services, Inc. 53p. 

49173716 Lawrence, J. (2013) Dose 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 by Eurofins-Agroscience Services, Inc. 55p. 

49173717 Lawrence, J. (2013) Dose Response Effects of MBI-203 DF and MBI-206 EP on the 

Parasitic Wasp (HYMENOPTERA: Aphelinidae Aphidius colemani) in a 

Translational Study in North Carolina: Final Report. Project Number: 

S13/00303/OCR. Unpublished study prepared by Eurofins-Agroscience Services, 

Inc. 52p. 

49173718 Lawrence, J. (2013) Dose Response Effects of MBI-203 DF and MBI-206 EP on the 

Mealybug Destroyer (COLEOPTERA: Coccinellidae Cryptolaemus montrouzieri) in 

a Translational Study in North Carolina: Final Report. Project Number: 

S13/00305/OCR. Unpublished study prepared by Eurofins-Agroscience Services, 

Inc. 53p. 

49173719 Lawrence, J. (2013) Dose Response Effects of MBI-203 DF and MBI-206 EP on the 

Predatory Spider Mite (MESOSTIGMATA: Phytoseiidae Amblyseius californicus) 

in a Translational Study in North Carolina: Final Report. Project Number: 

S13/00484/OCR. Unpublished study prepared by Eurofins-Agroscience Services, 

Inc. 54p. 

49173720 Lawrence, J. (2013) Dose Response Effects of MBI-203 DF and MBI-206 EP on the 

Sweet Potato Whitefly Predator (COLEOPTERA: Coccinellidae Delphastus 

catalinae) in a Translational Study in North Carolina: Final Report. Project Number: 

S13/00306/OCR. Unpublished study prepared by Eurofins-Agroscience Services, 

Inc. 53p. 

49173721 Lawrence, J. (2013) Dose Response Effects of MBI-203 DF and MBI-206 EP on the 
Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 24 of 33

Biopesticides Registration Action Document 

Aphid Midge (DIPTERA: Cecidomyiidae Aphidoletes aphidimyza) in a Translational 

Study in North Carolina: Final Report. Project Number: S13/00304/OCR. 

Unpublished study prepared by Eurofins-Agroscience Services, Inc. 50p. 

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

Application for Registration of MBI-206 EP. Transmittal of 11 Studies. 

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

EP on the Honey Bee (Hymenoptera: Apidae Apis mellifera) in a Laboratory Study 

in North Carolina. Project Number: S13/02866/OCR. Unpublished study prepared by 

Eurofins-Agroscience Services, Inc. 48p. 

49186302 Lawrence, J. (2013) Addendum to MRID # 491737-12 Contact Limit Test of MBI206 EP on the Honey Bee (Hymenoptera: Apidae Apis mellifera) in a Laboratory 

Study in North Carolina: Final Report. Project Number: S13/02867/OCR. 

Unpublished study prepared by Eurofins-Agroscience Services, Inc. 42p. 

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

MBI-203 DF and MBI-206 EP on the Parasitic Wasp (Hymenoptera: Aphelinidae 

Encarsia formosa) in a Translational Study in North Carolina: Final Report. Project 

Number: S13/00307/OCR. Unpublished study prepared by Eurofins-Agroscience 

Services, Inc. 61p. 

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

MBI-203 DF and MBI-206 EP on the Rove Beetle (Coleoptera: Staphylinidae Atheta 

coriaria) in a Translational Study in North Carolina. Project Number: 

S13/00301/OCR. Unpublished study prepared by Eurofins-Agroscience Services, 

Inc. 67p. 

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

MBI-203 DF and MBI-206 EP on the Bumble Bee (Hymenoptera: Apidae Bombus 

sp.) in a Translational Study in North Carolina. Project Number: S13/00300/OCR. 

Unpublished study prepared by Eurofins-Agroscience Services, Inc. 65p. 

49186306 Lawrence, J. (2013) Addendum to MRID # 491737-16 Dose 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 by Eurofins-Agroscience 

Services, Inc. 69p. 

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

MBI-203 DF and MBI-206 EP on Parasitic Wasp (Hymenoptera: Aphelinidae 

Aphidius colemani) in a Translational Study in North Carolina: Final Report. Project 

Number: S13/00303/OCR. Unpublished study prepared by Eurofins-Agroscience 

Services, Inc. 59p. 

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

MBI-203 DF and MBI-206 EP on the Mealybug Destroyer (Coleoptera: 

Coccinellidae Cryptolaemus montrouzieri) in a Translational Study in North 

Carolina: Final Report. Project Number: S13/00305/OCR. Unpublished study 

prepared by Eurofins-Agroscience Services, Inc. 61p. 

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

Biopesticides Registration Action Document 

MBI-203 DF and MBI-206 EP on the Predatory Spider Mite (Mesostigmata: 

Phytoseiidae Amblyseius swirskii) in a Translational 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 Dose Response Effects of 

MBI-203 DF and MBI-206 EP on the Sweet Potato Whitefly Predator (Coleoptera: 

Coccinellidae Delphastus catalinae) in a Translational 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 Dose Response Effects of 

MBI-203 DF and MBI-206 EP on the Aphid midge (Diptera: Cecidomyiidae 

Aphidoletes aphidimyza) in a Translational Study in North Carolina: Final Report. 

Project Number: S13/00304/OCR. Unpublished study prepared 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 Michael 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 Michael 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. Memorandum 

from Gail Tomimatsu, Ph.D. and Shannon Borges to Michael Glikes, dated December 13, 

2013. 

C. Other References 

Centers for Disease Control and Prevention. 2009. Drinking Water - Water Treatment. Available 

from http://www.cdc.gov/healthywater/drinking/public/water_treatment.html. 

DeFelice K, Wollenhaupt N, Buchholz D. 1993. Aquifers and Soil Filter Effect. Available from 

http://extension.missouri.edu/p/WQ24. 

U.S. EPA. 2004. Primer for Municipal Wastewater Treatment 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

Biopesticides Registration Action Document 

APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS 

(40 CFR PART 158 – SUBPART V) 

TABLE 1. Product Analysis Data Requirements for the 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 requirement for product identity. 482127-01 

487458-01 

885.1200 Manufacturing 

Process Submitted data fulfill the requirement for manufacturing 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 manufacturing process. 

482127-01 

487458-05 

487692-01 

487692-03 

TABLE 2. Product Analysis Data Requirements for the 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 manufacturing 

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 

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

Biopesticides Registration Action Document 

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 formula 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 additional 

information (confirmatory) on this data requirement. 

830.6319 Miscibility Non emulsifiable liquid. 

830.6320 Corrosion 

Characteristics 

As a term of registration, EPA is requiring additional 

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 additional 

information (confirmatory) on this data requirement. 

830.6319 Miscibility Miscible with water 

830.6320 Corrosion Characteristics As a term of registration, EPA is requiring additional 

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/mL

 
Heat-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 Active Ingredient Heat-killed Burkholderia spp. strain A396 cells and spent 

fermentation media and the End Use Pesticide Product 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 incidents, including 

immediate-type or delayed-type reactions of humans and domestic animals, 

occurred during research, development, or testing of 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 test note #4 of 40 CFR § 

158.2140(d)). 

N/A 

870.1100 Acute Oral Toxicity 

Oral LD50 female rats > 5,000 mg/kg body weight (bw) 

Classification: Acceptable 

TOXICITY CATEGORY IV

491737-01 

870.1200 Acute Dermal Toxicity

Dermal LD50 combined (female rats) > 5,050 mg/kg bw 

Classification: Acceptable 

TOXICITY CATEGORY IV

491737-02 

870.1300 Acute Inhalation 

Toxicity 

LC50 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 the 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 the 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

Biopesticides Registration Action Document 

TABLE 6. Nontarget Organism Toxicity and Environmental 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 

LD50 (conducted on 23-day old chicks) 

 was determined > 112.5 mg a.i./kg bw. 

Classification: Supplemental 

48212711 

48831001 

49173700 

LD50 >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. LD50 >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 LC50 

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 LC50 > 100 mg MBI-206EP/L). 

Classification: Supplemental 

48625801 

48831001 

49173700 

NOEC and EC50 > 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 

LC50 was determined to be > 81 mg a.i. /L. 

Classification: Supplemental 

48212713 

48831001 

48745803 

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

Biopesticides Registration Action Document 

TABLE 6. Nontarget Organism Toxicity and Environmental 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 EC50 = 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 

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

Biopesticides Registration Action Document 

TABLE 6. Nontarget Organism Toxicity and Environmental 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

49173718 
Heat-killed Burkholderia sp. strain A396 Cells and Spent Fermentation Media Page 32 of 33

Biopesticides Registration Action Document 

TABLE 6. Nontarget Organism Toxicity and Environmental 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