BIOPESTICIDES REGISTRATION ACTION DOCUMENT

Chromobacterium subtsugae strain PRAA4-1T

Pesticide Chemical (PC) Code: 016329

U.S. Environmental Protection Agency

Office of Pesticide Programs

Biopesticides and Pollution Prevention Division

September 27, 2011
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TABLE OF CONTENTS

I. EXECUTIVE SUMMARY .............................................................................................. 4

II. ACTIVE INGREDIENT OVERVIEW........................................................................... 6

III. REGULATORY BACKGROUND.................................................................................. 6

 A. Applications for Pesticide Product Registration............................................................................6

 B. Food Tolerance Exemption..............................................................................................................6

IV. RISK ASSESSMENT ....................................................................................................... 7

 A. Product Analysis Assessment (40 CFR § 158.2120).......................................................................7

 B. Human Health Assessment (40 CFR § 158.2140)...........................................................................7

 C. Environmental Assessment (40 CFR § 158.2150) ........................................................................13

V. ENVIRONMENTAL JUSTICE.................................................................................... 17

VI. RISK MANAGEMENT DECISION............................................................................. 18

VII. ACTIONS REQUIRED BY THE REGISTRANT ...................................................... 18

 A. Final Printed Labeling ...................................................................................................................18

 B. Terms of Registration.....................................................................................................................18

 C. Reporting of Adverse Effects and Hypersensitivity Incidents....................................................19

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS ......................................... 20

IX. BIBLIOGRAPHY........................................................................................................... 21

A. Studies Submitted to Support the Chromobacterium subtsugae strain PRAA4-1T 

 Pesticide Product Registrations.....................................................................................................21

B. Environmental Protection Agency Risk Assessment Memoranda and Other Documents.......24

C. Other References.............................................................................................................................24

APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS............................ 26

APPENDIX B. PESTICIDE PRODUCTS............................................................................... 36
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BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

Office of Pesticide Programs (OPP)

Biopesticides and Pollution Prevention Division

Microbial Pesticides Branch

Science Reviews

Joel V. Gagliardi, Ph.D. Product Analysis, Human Health

John L. Kough, Ph.D. Product Analysis, Human Health

Anna Gross Environmental Effects

Zigfridas Vaituzis, Ph.D. Environmental Effects

Shannon Borges Environmental Effects

Regulations

Sheryl K. Reilly, Ph.D. Chief, Microbial Pesticides Branch

Jeannine Kausch Regulatory Action Leader
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I. EXECUTIVE SUMMARY

Chromobacterium subtsugae strain PRAA4-1T is a gram-negative, violet-pigmented bacterium 

that was isolated from soil under an eastern hemlock (Tsuga canadensis) in the Catoctin 

Mountain region of central Maryland. The United States Department of Agriculture found this 

isolate of Chromobacterium subtsugae to be orally toxic to Colorado potato beetle (Leptinotarsa 

decemlineata) larvae, small hive beetle (Aethina tumida) larvae, southern corn rootworm 

(Diabrotica undecimpunctata) larvae and adults, and southern green stink bug (Nezara viridula)

adults. Additional testing has shown that Chromobacterium subtsugae strain PRAA4-1T-treated 

diet resulted in reduced feeding in beet armyworm (Spodoptera exigua), cabbage looper 

(Trichoplusia ni), tobacco budworm (Heliothis virescens), diamondback moth (Plutella 

xylostella), and southern corn rootworm, suggesting this microbe’s insecticidal activity is due to 

reduction in weight or inhibition of feeding. In light of the demonstrated insecticidal and 

miticidal capabilities of Chromobacterium subtsugae strain PRAA4-1T, Marrone Bio Innovations 

proposed to register both a manufacturing-use pesticide product, MBI-203 TGAI, and an end-use 

pesticide product, MBI-203 EP Bioinsecticide (formerly MBI-203 EP), containing this 

bacterium. MBI-203 EP Bioinsecticide will be used to control label-specified insect and mite 

pests (e.g., European corn borer, citrus rust mite, and stink bugs) on agricultural and greenhouse

crops, including vegetables, fruit, flowers, bedding plants, ornamentals, and turf. 

Environmental Protection Agency (EPA) scientists reviewed product analysis, toxicology, and 

nontarget organism data and information (40 Code of Federal Regulations (CFR) §§ 158.2120, 

158.2140, and 158.2150, respectively) submitted to support the registration of the two 

aforementioned Chromobacterium subtsugae strain PRAA4-1T pesticide products. Overall, such 

data and information are adequate for risk assessment purposes, fulfill current data requirements,

and allow for registration under Section 3(c)(5) of the Federal Insecticide, Fungicide, and 

Rodenticide Act (FIFRA).

Product analysis data requirements for Chromobacterium subtsugae strain PRAA4-1T

, including 

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

characteristics, were fulfilled by acceptable guideline studies. 

Adequate mammalian toxicology data and information were submitted to support the 

Chromobacterium subtsugae strain PRAA4-1T pesticide products. The acute injection 

toxicity/pathogenicity study showed that, at a single high dose, Chromobacterium subtsugae 

strain PRAA4-1T is not toxic, infective, and/or pathogenic via the intravenous route of exposure.

Acute pulmonary and oral toxicity/pathogenicity tests were waived after considering the results 

of the intravenous test and MBI-203 TGAI-specific toxicity tests (acute oral and inhalation).

Moreover, no hypersensitivity incidents, occurring during research, development, or testing of 

Chromobacterium subtsugae strain PRAA4-1T, were reported by the applicant. In light of the 

results of the intravenous study, the acceptable rationale provided to waive the acute pulmonary 

and oral toxicity/pathogenicity tests, and the absence of hypersensitivity incidents, testing at 

higher tiers (i.e., Tiers II and III) was not required.

EPA concluded that there is a reasonable certainty that no harm will result to the United States 

population, including infants and children, from aggregate exposure to residues of 

Chromobacterium subtsugae strain PRAA4-1T

. No dietary risks are expected from use of 

Chromobacterium subtsugae strain PRAA4-1Tas an active ingredient in pesticide products. 
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Significant dietary exposure—through food and drinking water—is not anticipated given the

potential for this bacterium to degrade due to both predation by other biological organisms and

exposure to environmental factors (e.g., sunlight and varying temperatures) in the phyllosphere, 

the filtering effect of many particulate soil types, and conditions (e.g., filtration and pH 

adjustments) water is subjected to in wastewater treatment systems and drinking water facilities. 

Should Chromobacterium subtsugae strain PRAA4-1T be present on food or in drinking water, 

the acute oral toxicity and pathogenicity data/information demonstrated no toxicity, infectivity, 

and/or pathogenicity is likely to occur with any exposure level of this microbial pest control 

agent.

Despite the low toxicological profile of Chromobacterium subtsugae strain PRAA4-1T, baseline 

personal protective equipment (PPE) is required for handlers that may be exposed to the active 

ingredient, due to their occupation, for prolonged periods or numerous times. Handlers working 

with Chromobacterium subtsugae strain PRAA4-1T in agricultural settings are directed to wear a 

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

meeting National Institute for Occupational Safety and Health (NIOSH) standards of at least N95, R-95, or P-95. EPA may require additional PPE, other than the standard described above, on 

a product-specific basis. 

The data and data waiver rationale, submitted by the applicant to support the pesticide products 

containing Chromobacterium subtsugae strain PRAA4-1T

, are sufficient to fulfill the Tier I 

nontarget organism data requirements and for risk assessment purposes. Further testing of 

nontarget organisms at higher tier levels (i.e., Tiers II, III, and IV) is not required for the current 

uses and application methods. EPA performed an environmental risk assessment based on the 

data and data waiver rationale provided by the applicant and determined that the uses of 

Chromobacterium subtsugae strain PRAA4-1T do not pose significant risk to nontarget 

organisms when used according to label directions. Based on study results, there are concerns of 

toxicity to terrestrial arthropods, aquatic invertebrates, and honey bees. Use sites and application 

methods, in combination with mitigating label language (see Table 7 in Appendix A), however, 

will limit exposure. EPA has made “no effect” determinations for direct effects, indirect effects, 

and effects to habitat (including designated critical habitat) to listed species for all foliar and soildirected applications to crop plants treated within greenhouses. On the other hand, EPA 

concludes that all outdoor foliar applications made by chemigation and aerial spray equipment 

have the potential to cause direct effects, indirect effects, and effects to habitat (including 

designated critical habitat) to listed species. 

On October 1, 2009, EPA announced a new policy to provide a more meaningful opportunity for 

the public to participate in major registration decisions before they occur. According to this 

policy, 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 EPA believes there 

may be significant public interest.

 

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

containing Chromobacterium subtsugae strain PRAA4-1T

, a new active ingredient, and allowing 

for this active ingredient’s first outdoor and food uses were subject to a 30-day comment period. 

During this comment period, no comments were received on EPA’s preliminary decision to 

register the Chromobacterium subtsugae strain PRAA4-1T pesticide products, MBI-203 TGAI 
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and MBI-203 EP Bioinsecticide. Therefore, EPA maintained that, based upon the risk assessment 

and information submitted in support of registration of such pesticide products, it was in the best 

interest of the public and the environment to issue the MBI-203 TGAI and MBI-203 EP

Bioinsecticide registrations. The basis for this decision can be found in the risk assessment for

Chromobacterium subtsugae strain PRAA4-1T, which is characterized throughout this BRAD. 

II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Chromobacterium subtsugae strain PRAA4-1T

Culture Deposit: Agricultural Research Service Culture Collection (also 

known as the Northern Regional Research Laboratory 

(NRRL) Collection) in Peoria, Illinois under Accession 

Number NRRL B-30655

OPP Chemical Code: 016329

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 registered pesticide products containing this 

active ingredient.

III. REGULATORY BACKGROUND

A. Applications for Pesticide Product Registration

On December 22, 2009, Marrone Bio Innovations (address: 2121 Second Street, Suite B-107; 

Davis, California 95618) submitted applications to register a manufacturing-use pesticide

product, MBI-203 TGAI (EPA File Symbol 84059-O), and an end-use pesticide product, MBI203 EP (EPA File Symbol 84059-RN), under FIFRA section 3. On March 10, 2010, EPA

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

ingredient (75 Federal Register (FR) 11175) and opened a 30-day public comment period 

pursuant to the provisions of FIFRA section 3(c)(4). No comments were received following this 

publication. 

B. Food Tolerance Exemption

Concurrent with its registration applications and under Federal Food, Drug, and Cosmetic Act 

(FFDCA) section 408(d), Marrone Bio Innovations submitted a petition to establish an 

exemption from the requirement of a tolerance for Chromobacterium subtsugae strain PRAA4-1T 

(Pesticide Petition (PP) 9F7674). In the Federal Register of March 10, 2010 (75 FR 11171), EPA

announced that Marrone Bio Innovations proposed to establish an exemption from the 

requirement of a tolerance for residues of the insecticide, Chromobacterium subtsugae strain 
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PRAA4-1T, in or on all food commodities and opened a 30-day comment period. No comments 

were received following this publication. 

On September 7, 2011, EPA established an exemption from the requirement of a tolerance for 

residues of Chromobacterium subtsugae strain PRAA4-1T in or on all food commodities when 

applied as an insecticide or miticide and used in accordance with good agricultural practices (40 

CFR § 180.1305; 76 FR 55272).

IV. RISK ASSESSMENT

In the Federal Register of October 26, 2007, EPA issued a Final Rule on the data requirements to 

support registration of microbial pesticides and updated the definition for microbial pesticides 

(72 FR 61002). The rule became effective on December 26, 2007. The data and information 

evaluated for this BRAD were considered in light of these requirements.

The classifications that are found for each data submission are assigned by EPA science 

reviewers and are an indication of the usefulness of the information contained in the documents 

for risk assessment. A rating of “acceptable” indicates the study is scientifically sound and is 

useful for risk assessment. A “supplemental” rating indicates the data provide some information 

that can be useful for risk assessment. The studies may have certain aspects determined not to be 

scientifically acceptable (“supplemental: 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 EPA in support of a 

pesticide registration. The active ingredient or particular product is classified into Toxicity 

Category I, II, III, or IV, where Toxicity Category I indicates the highest toxicity and Toxicity 

Category IV indicates the lowest toxicity (see 40 CFR § 156.62). 

A. Product Analysis Assessment (40 CFR § 158.2120)

All product analysis data requirements for Chromobacterium subtsugae strain PRAA4-1T have 

been fulfilled. Refer to Tables 1, 2, 3, and 4 in Appendix A for a brief 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 Chromobacterium subtsugae strain PRAA4-1T have been 

fulfilled. Acceptable Tier I mammalian toxicology data and information support registration of

the Chromobacterium subtsugae strain PRAA4-1T pesticide products. Furthermore, Tier II and 

Tier III studies were not required for Chromobacterium subtsugae strain PRAA4-1T based on the

lack of acute toxicity/pathogenicity in the Tier I studies.
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For a comprehensive summary of the generic toxicology data requirements described in sections 

IV(B)(1)(a) and IV(B)(1)(b), as well as additional product-specific data submitted to support the 

individual registrations, refer to Tables 5 and 6 in Appendix A.

a. Acute Toxicity/Pathogenicity – Tier I

Acute Oral Toxicity/Pathogenicity (Office of Chemical Safety and Pollution Prevention (OCSPP) 

Guideline 885.3050; Master Record Identification Number (MRID No.) 479450-23): Upon 

consideration of results of other definitive toxicological data submitted by the applicant, EPA 

waived acute oral toxicity/pathogenicity testing for Chromobacterium subtsugae strain PRAA4-

1T. In an acute up and down oral toxicity study (MRID No. 479450-03), three female rats were 

gavaged once with 5,000 milligrams per kilogram (mg/kg) of the test material. All animals 

survived and gained weight, and there were no signs of gross toxicity, adverse effects, or 

abnormal behavior. The acute oral median lethal dose (LD50) was greater than 5,000 mg/kg. In an 

acute intravenous study (MRID No. 479450-11), groups of rats received a single intravenous 

injection of inactive test material or active test material (2.62 x 108 colony-forming units per 

milliliter (cfu/mL)). Three males and three females from the active test material group were 

sacrificed and necropsied on days 0, 3, 7, 14, and 21 after treatment. Tissue and blood samples 

were collected at necropsy and cultured to provide qualitative and quantitative measurements of 

the test organism. There was no mortality during the study, and all rats appeared normal 

throughout. The test organism had cleared completely from blood, kidneys, mesenteric lymph 

nodes, lungs, brain, liver, spleen, and cecum contents by day 3. The test material was nontoxic 

and noninfective. EPA believes these data, when taken together, clearly indicate that 

Chromobacterium subtsugae strain PRAA4-1T would not be toxic, infective, and/or pathogenic 

through the oral route of exposure and that further testing is not necessary.

Acute Pulmonary Toxicity/Pathogenicity (OCSPP Guideline 885.3150; MRID No. 479450-23): 

Upon consideration of results of other definitive toxicological data submitted by the applicant, 

EPA waived acute pulmonary toxicity/pathogenicity testing for Chromobacterium subtsugae

strain PRAA4-1T. In an acute inhalation study (MRID No. 479450-05), male and female rats 

were exposed to an aerosol concentration of 2.12 milligrams per liter (mg/L) of the test material 

for 4 hours in a nose-only inhalation chamber. No deaths occurred during exposure or during the 

14-day observation period. Clinical signs included decreased activity and piloerection, which 

resolved by day 5. Necropsy was unremarkable. The acute inhalation median lethal concentration 

(LC50) was greater than 2.12 mg/L. In an acute intravenous study (MRID No. 479450-11), groups 

of rats received a single intravenous injection of inactive test material or active test material (2.62 

x 108 cfu/mL). Three males and three females from the active test material group were sacrificed 

and necropsied on days 0, 3, 7, 14, and 21 after treatment. Tissue and blood samples were 

collected at necropsy and cultured to provide qualitative and quantitative measurements of the 

test organism. There was no mortality during the study, and all rats appeared normal throughout. 

The test organism had cleared completely from blood, kidneys, mesenteric lymph nodes, lungs, 

brain, liver, spleen, and cecum contents by day 3. The test material was nontoxic and 

noninfective. EPA believes these data, when taken together, clearly indicate that 

Chromobacterium subtsugae strain PRAA4-1T would not be toxic, infective, and/or pathogenic 

through the inhalation route of exposure and that further testing is not necessary.
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Acute Injection Toxicity/Pathogenicity (Intravenous) – Rat (OCSPP Guideline 885.3200; MRID 

No. 479450-11): In an acute intravenous injection toxicity and pathogenicity study, groups of 

young adult Sprague-Dawley rats were injected at 3.1 x 106 colony-forming units (cfu)/animal.

Animals were observed up to 21 days. Five males and five females were treated with inactive 

MOI 203 [MBI 203] TGAI, while five males and five females were not treated and served as 

controls. All animals survived, gained weight, and appeared normal during the study. No 

observable abnormalities were noted in any animal at necropsy. There were no significant 

variations among organ weights. MOI 203 [MBI 203] organism cleared from the blood, brain, 

lungs, spleen, liver, kidneys, mesenteric lymph nodes, and cecum contents of animals by day 3. 

MOI 203 [MBI 203] was not detected in the tissues/organs of animals treated with autoclaved 

test material or the untreated control group. MOI 203 [MBI 203] does not appear to be toxic, 

infective, and/or pathogenic to rats when dosed intravenously at 3.1 x 106 cfu per animal. This 

study was rated acceptable.

Hypersensitivity Incidents (OCSPP Guideline 885.3400; MRID No. 479450-23): No 

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

domestic animals that occurred during research, development, or testing of Chromobacterium 

subtsugae strain PRAA4-1T, were reported by the applicant. Any future hypersensitivity incidents 

must be reported to EPA (refer to test note #3 of 40 CFR § 158.2140(d)).

Cell Culture (OCSPP Guideline 885.3500): This study is not required because Chromobacterium 

subtsugae strain PRAA4-1T is not a virus (refer to test note #4 of 40 CFR § 158.2140(d)).

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

 Reproductive Fertility Effects, Carcinogenicity, Immunotoxicity, and 

 Infectivity/Pathogenicity Analysis – Tier III

Tier II and Tier III studies were not required for Chromobacterium subtsugae strain PRAA4-1T

based on the lack of acute toxicity/pathogenicity in the Tier I studies.

c. Endocrine Disruptors

As required under FFDCA section 408(p), EPA has developed the Endocrine Disruptor 

Screening Program (EDSP) 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.” The EDSP employs a two-tiered approach to making the statutorily required 

determinations. Tier 1 consists of a battery of 11 screening assays to identify the potential of a 

chemical substance to interact with the estrogen, androgen, or thyroid (E, A, or T) hormonal 

systems. Chemicals that go through Tier 1 screening and are found to have the potential to 

interact with E, A, or T hormonal systems will proceed to the next stage of the EDSP where EPA 

will determine which, if any, of the Tier 2 tests are necessary based on the available data. Tier 2 

testing is designed to identify any adverse endocrine-related effects caused by the substance, and 

establish a quantitative relationship between the dose and the E, A, or T effect.

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. This list 

of chemicals was selected based on the potential for human exposure through pathways such as 
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food and water, residential activity, and certain post-application agricultural scenarios. This list 

should not be construed as a list of known or likely endocrine disruptors.

Chromobacterium subtsugae strain PRAA4-1T is not among the group of 58 pesticide active 

ingredients on the initial list to be screened under the EDSP. Under FFDCA section 408(p), EPA 

must screen all pesticide chemicals. Accordingly, EPA anticipates issuing future EDSP 

orders/data call-ins for all pesticide active ingredients. 

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 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 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, EPA must take into account the factors set forth in section 408(b)(2)(C) of FFDCA, 

which require EPA to give special consideration to exposure of infants and children to the 

pesticide chemical residue in establishing a tolerance and to “ensure that there is a reasonable 

certainty that no harm will result to infants and children from aggregate exposure to the pesticide 

chemical residue....” Additionally, section 408(b)(2)(D) of FFDCA requires that EPA consider 

“available information concerning the cumulative effects of [a particular pesticide's] . . . residues 

and other substances that have a common mechanism of toxicity.”

EPA performs a number of analyses to determine the risks from aggregate exposure to pesticide 

residues. First, EPA determines the toxicity of pesticides. Second, 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, 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. EPA has 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 

Chromobacterium subtsugae strain PRAA4-1T have been fulfilled.
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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: Any exposure to this naturally occurring soil 

bacterium is anticipated to be negligible. Although Chromobacterium subtsugae strain PRAA4-

1T may be applied directly to food, it is not expected to persist or accumulate in any reservoirs on 

plants or food commodities (the phyllosphere) because, as a soil microorganism, it is best 

adapted to more favorable conditions underground. Rather, after application, it likely will 

degrade due to predation by other biological organisms (e.g., protists) and exposure to particular 

environmental factors (e.g., sunlight and varying temperatures) (Lindow and Brandl 2003; U.S. 

EPA 1996). Should this microbial pesticide be present on food, the acute oral toxicity and 

pathogenicity data/information demonstrated no toxicity, infectivity, and/or pathogenicity is 

likely to occur with any exposure level of Chromobacterium subtsugae strain PRAA4-1T (see 

section IV(B)(1)(a) and Table 5 in Appendix A).

Drinking Water Exposure and Risk Characterization: Exposure of humans to residues of 

Chromobacterium subtsugae strain PRAA4-1T in consumed drinking water is unlikely. The

proposed use patterns for Chromobacterium subtsugae strain PRAA4-1T do not include direct 

application to aquatic environments, thereby limiting contact with surface water. Furthermore,

ground water is not expected to have significant exposure to Chromobacterium subtsugae strain 

PRAA4-1T since, like other microorganisms, this bacterium would likely be filtered out by the 

particulate nature of many soil types (Aislabie et al. 2001; DeFelice et al. 1993; Pang et al. 2008) 

and is not known to survive in water or deep soil. If Chromobacterium subtsugae strain PRAA4-

1T 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 high temperatures, chlorination, pH adjustments, and/or filtration

(Centers for Disease Control and Prevention 2009; U.S. EPA 2004). In the remote likelihood that 

this microbial pesticide is present in drinking water (e.g., in water not subject to treatment 

systems or facilities), the acute oral toxicity and pathogenicity data/information demonstrated no 

toxicity, infectivity, and/or pathogenicity is likely to occur with any exposure level of 

Chromobacterium subtsugae strain PRAA4-1T (see section IV(B)(1)(a) and Table 5 in Appendix 

A).

Non-occupational, Residential Risk Characterization: Dermal and inhalation non-occupational 

exposure to Chromobacterium subtsugae strain PRAA4-1T is not expected as all proposed 

pesticide applications will take place in distinct agricultural settings. Even if dermal and 

inhalation non-occupational exposures were to occur, such exposures would not exceed EPA’s 

level of concern given testing that indicated that Chromobacterium subtsugae strain PRAA4-1T

is not toxic (acute inhalation and dermal toxicity), is only slightly irritating (primary dermal 

irritation), is not a sensitizer (dermal sensitization), and is not pathogenic or infective (acute 

injection toxicity/pathogenicity) (see section IV(B)(1)(a) and Table 5 in Appendix A).
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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, EPA consider “available information concerning the cumulative 

effects of [a particular pesticide’s] . . . residues and other substances that have a common 

mechanism of toxicity.”

EPA has not found Chromobacterium subtsugae strain PRAA4-1T to share a common 

mechanism of toxicity with any other substances, and Chromobacterium subtsugae strain 

PRAA4-1T does not appear to produce a toxic metabolite produced by other substances. For the 

purposes of this tolerance action, therefore, EPA has assumed that Chromobacterium subtsugae

strain PRAA4-1T does not have a common mechanism of toxicity with other substances. 

Following from this, therefore, EPA concludes that there are no cumulative effects associated 

with Chromobacterium subtsugae strain PRAA4-1T that need to be considered. For information 

regarding EPA’s efforts to determine which chemicals have a common mechanism of toxicity 

and to evaluate the cumulative effects of such chemicals, see EPA’s website at 

http://www.epa.gov/pesticides/cumulative.

c. Determination of Safety for the United States (U.S.) Population, Infants and 

 Children

FFDCA section 408(b)(2)(C) provides that 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 EPA shall apply an additional tenfold (10X) margin of 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 EPA determines that a 

different margin of safety will be safe for infants and children. This additional margin of safety is 

commonly referred to as the Food Quality Protection Act Safety Factor. In applying this 

provision, EPA either retains the default value of 10X or uses a different additional safety factor 

when reliable data available to EPA support the choice of a different factor.

Based on the acute toxicity and pathogenicity data/information summarized in section 

IV(B)(1)(a) and Table 5 in Appendix A, EPA concludes that there is a reasonable certainty that 

no harm will result to the U.S. population, including infants and children, from aggregate 

exposure to the residues of Chromobacterium subtsugae strain PRAA4-1T

. Such exposure 

includes all anticipated dietary exposures and all other exposures for which there is reliable 

information. EPA has arrived at this conclusion because, considered collectively, the data (e.g., 

lack of toxicity noted for oral, dermal, and inhalation routes of exposure) available on 

Chromobacterium subtsugae strain PRAA4-1T do not demonstrate toxic, pathogenic, and/or 

infective potential to sensitive populations from exposure to this microbial pest control agent. 

Thus, there are no threshold effects of concern and, as a result, the provision requiring an 

additional margin of safety is not necessary. 
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3. Occupational Exposure and Risk Characterization

Handler exposure to Chromobacterium subtsugae strain PRAA4-1T is not expected to pose any 

undue risk. Regardless, appropriate personal protective equipment and precautionary statements 

are required on pesticide product labels to mitigate any potential risks to pesticide handlers due to 

prolonged or numerous exposures. Handlers applying Chromobacterium subtsugae strain 

PRAA4-1T end-use pesticide products in agricultural settings must wear a long-sleeved shirt, 

long pants, socks, shoes, waterproof gloves, and a dust/mist filtering respirator meeting NIOSH 

standards of at least N-95, R-95, or P-95. Additional PPE, other than the standard described 

above, may be required on a product-specific basis.

4. Human Health Risk Characterization

EPA considered human exposure to Chromobacterium subtsugae strain PRAA4-1T in light of the 

standard for registration in FIFRA and the relevant safety factors in FFDCA. A determination has 

been made that no unreasonable adverse effects to the U.S. population in general, and to infants 

and children in particular, will result when Chromobacterium subtsugae strain PRAA4-1T

pesticide products are used in accordance with EPA-approved labeling.

C. Environmental Assessment (40 CFR § 158.2150)

The data and data waiver rationale, submitted by the applicant to support the pesticide products 

containing Chromobacterium subtsugae strain PRAA4-1T, fulfill the Tier I nontarget organism 

data requirements and are sufficient for risk assessment purposes. Further testing of nontarget 

organisms at higher tier levels (i.e., Tiers II, III, and IV) is not required for the current uses and 

application methods. EPA has performed an environmental risk assessment based on the data and 

data waiver rationale provided by the applicant and has determined that the use of 

Chromobacterium subtsugae strain PRAA4-1T does not pose significant risk to nontarget 

organisms when used according to label directions. Based on study results, there are concerns of 

toxicity to terrestrial arthropods, aquatic invertebrates, and honey bees. Use sites and application 

methods, in combination with mitigating label language, however, will limit exposure.

For a comprehensive summary of the generic data requirements described in sections IV(C)(1), 

refer to Table 7 in Appendix A.

1. Ecological Exposure and Risk Characterization 

a. Terrestrial Animals and Plants

The end-use product (EP) is a concentrated liquid that is labeled as a foliar spray with ground or 

aerial applications at 3-10 day intervals. Aerial applications would increase exposure to nontarget 

insects and honey bees and, due to potential toxicity concerns, were not recommended.

Terrestrial animals and plants may be exposed to Chromobacterium subtsugae strain PRAA4-1 T

through direct sprays and exposure on foliar surfaces. 

Birds (OCSPP Guideline 885.4050) and Mammals (OCSPP Guideline 885.4150): A study 

(MRID No. 479450-13) was conducted to determine the acute toxicity and pathogenicity of MBI203 TGAI (active ingredient, 100.00% Chromobacterium subtsugae strain PRAA4-1T cells and 
Chromobacterium subtsugae strain PRAA4-1T Page 14 of 36

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spent fermentation media) to northern bobwhite (Colinus virginianus). Fourteen-day-old chicks 

were dosed with the test material (4.0 x 1011 colony-forming units per kilogram (cfu/kg) body 

weight), attenuated test material, or sterile saline once a day for five days and observed for an 

additional 25 days. No signs of toxic or pathogenic effects were observed throughout the test or 

at necropsy. This study demonstrates that toxicity/pathogenicity to birds is not expected from 

labeled applications of Chromobacterium subtsugae strain PRAA4-1T.

In an acute oral toxicity study (MRID No. 479450-03), three fasted young adult female SpragueDawley rats were given a single oral gavage dose of MOI 203 [MBI 203] TGAI at a

concentration of 5,000 mg/kg body weight. The test material was allowed to thaw at room 

temperature prior to dosing. The animals were observed for 14 days. All animals survived, 

gained weight, and appeared active and healthy throughout the study. No gross abnormalities 

were found in any animal at necropsy. This study demonstrates that toxicity/pathogenicity to wild 

mammals is not expected from labeled applications of Chromobacterium subtsugae strain 

PRAA4-1T.

Nontarget Insects (OCSPP Guideline 885.4340) and Honey Bees (OCSPP Guideline 885.4380):

Nontarget insect studies were submitted for ladybird beetles, green lacewing larvae and parasitic 

wasps. Two studies were submitted for ladybird beetles, both of which indicated toxicity. An 

acute toxicity study (MRID No. 479450-16) was of insufficient duration but supplemental for the 

purposes of the risk assessment, and a feeding study (MRID No. 479450-17) fulfilled the testing 

guidelines but also indicated toxicity. A green lacewing study (MRID No. 479450-18) that 

fulfilled testing requirements was submitted; there were no observed effects from test 

concentrations of Chromobacterium subtsugae strain PRAA4-1T, and the LC50 for MBI TGAI 

was greater than 70,000 parts per million (ppm) in a moth egg diet. A parasitic wasp study 

(MRID No. 481129-05) was also submitted and was supplemental for the purposes of the risk 

assessment. While there were no statistically significant differences in the control and test 

substance groups, the dose of Chromobacterium subtsugae strain PRAA4-1T to which the wasps 

were exposed was not adequately characterized and the duration of the test was insufficient.

Three honey bee studies were submitted, all of which were sufficient for the purposes of the risk 

assessment. Results indicated toxicity to honey bees with the use of Chromobacterium subtsugae

strain PRAA4-1T (MRID Nos. 479450-19, 479450-20 and 479450-21).

In conclusion, according to studies conducted on nontarget insects and honey bees, there are 

toxicity concerns with the use of Chromobacterium subtsugae strain PRAA4-1T. To address 

these concerns and to reduce exposure to nontarget insects and honey bees that may occur 

through outdoor ground and aerial applications, EPA is requiring particular aerial drift reduction 

information, as well as the following mitigation language, on the MBI-203 EP Bioinsecticide

label: 

Environmental Hazards Statements

This product is highly toxic to bees exposed to direct treatment or residues on blooming crops or 

weeds. Do not apply this product or allow it to drift to blooming crops or weeds if bees are 

visiting the treatment area.
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This product is toxic to certain nontarget terrestrial arthropods. Minimize spray drift away from 

target area to reduce effects to nontarget insects.

Nontarget Plants (OCSPP Guideline 885.4300): The applicant submitted both data waiver 

rationale (MRID No. 479450-23) and a study (MRID No. 479450-15) to fulfill the nontarget 

plant testing requirement. While Chromobacterium subtsugae strain PRAA4-1T occurs naturally 

in soils and is not related to any known plant pathogen, effects of mild phytotoxicity were 

observed in cucumbers at 5X and 10X the field application rate, and in chrysanthemum flowers 

at all application rates. Even though effects in cucumbers and chrysanthemum flowers were mild, 

the MBI-203 EP Bioinsecticide label will still require all users to test for phytotoxicity prior to 

pesticide application.

b. Aquatic Animals and Plants

Freshwater Fish (OCSPP Guideline 885.4200), Freshwater Invertebrates (OCSPP Guideline 

885.4240), and Estuarine/Marine Fish and Invertebrates (OCSPP Guideline 885.4280): MBI203 EP Bioinsecticide (94.50% Chromobacterium subtsugae strain PRAA4-1T) is not labeled for 

aquatic uses, and the label includes language to prevent application to or contamination of water. 

Due to demonstrated toxicity to aquatic invertebrates, aquatic uses should not be added to the 

label without further testing. The applicant submitted studies to fulfill the freshwater fish and 

freshwater invertebrate testing requirements, as well as data waiver rationale for testing on 

estuarine/marine organisms. The freshwater fish study (MRID No. 479450-14) fulfills the testing 

requirements and did not show any adverse effects to fathead minnows (Pimephales promelas). 

Toxicity was shown in two supplemental studies (MRID Nos. 479450-12 and 481129-04) 

submitted by the applicant that tested the effects of Chromobacterium subtsugae strain 

PRAA4-1T on Daphnia magna. The studies were sufficient for the purposes of this registration, 

but do not support aquatic uses. The data waiver rationale submitted for estuarine/marine 

organisms was acceptable for the labeled use patterns, as MBI-203 EP Bioinsecticide is not 

intended for direct application into the estuarine or marine environment or expected to enter such 

environment in significant concentrations.

Even though MBI-203 EP Bioinsecticide is not applied directly to aquatic environments, EPA is 

requiring that the maximum labeled application rate not exceed 12 quarts of product per acre and 

that specific aerial drift reduction text, as well the following mitigation language, be included on 

the label to reduce the potential for exposure to aquatic invertebrates: 

Environmental Hazards Statements

This product is toxic to aquatic invertebrates. Drift and runoff may be hazardous to aquatic 

organisms in water adjacent to treated areas.

Buffer Zone for Aerial Applications (in the Directions for Use) (see U.S. EPA 2011c, 2011d)

Do not apply within 75 feet of aquatic habitats (such as, but not limited to, lakes, reservoirs, 

rivers, streams, marshes, ponds, estuaries, and commercial fish ponds).
Chromobacterium subtsugae strain PRAA4-1T Page 16 of 36

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Information on Droplet Size (in the Directions for Use) (see U.S. EPA 2011c, 2011d)

Use only medium or coarser spray nozzles according to ASAE (S572) definition for standard 

nozzles.

c. Open Literature Information

Literature searches were performed to determine if other effects have been reported that were not 

contained within the information provided by the applicant. Both the Science Citation Index 

Expanded database (1970-present) and the Environmental Information Search service (1964-

present) were searched using the terms “Chromobacterium” and “Chromobacterium subtsugae.”

The Environmental Information Search service includes the Agricola; Biosis Previews; CAB 

Abstracts; Energy, Science, and Technology; General Science Abstracts; National Technical 

Information Service; and Waternet databases. None of the records found in any of the searches 

described here reported effects on any nontarget species. The only results relevant to the active 

ingredient were two documents published by the United States Department of Agriculture –

Agricultural Research Service concerning the toxicity of Chromobacterium subtsugae to 

Colorado potato beetle and other insect pests (Martin et al. 2007a), as well as to southern green 

stink bug and corn rootworm (Martin et al. 2007b). Neither document provided additional 

information on environmental or nontarget effects useful for the purposes of this risk assessment. 

2. Environmental Fate Data 

As the information provided, for the current uses and application methods, is sufficient to both

satisfy the Tier I nontarget organism data requirements and for nontarget organism risk 

assessment for Chromobacterium subtsugae strain PRAA4-1T, further testing at higher tier levels

(i.e., Tiers II, III, and IV) is not required. 

3. Threatened and Endangered Species Assessment

EPA has determined that there is a potential for effects to nontarget organisms, including 

endangered and threatened species as a result of the proposed labeled applications (aerial sprays 

and chemigation) of Chromobacterium subtsugae strain PRAA4-1T. There are toxicity concerns 

for certain terrestrial arthropods and honey bees, as well as for aquatic invertebrates. Further 

analysis is needed to characterize the specific effects and affected species. For this reason, aerial 

applications were not recommended. Terrestrial applications through sprinkler irrigation systems 

(chemigation) are expected to result in exposure to nontarget insects, though to a lesser extent 

than aerial applications. MBI-203 EP Bioinsecticide is also labeled for greenhouse use, which 

will not result in significant exposure to nontarget organisms. Aquatic applications are not 

permitted for this product, which will limit exposure to aquatic invertebrates and other aquatic 

organisms. Mitigation language (e.g., requirement for a buffer between aquatic and treatment 

areas and use of only medium to coarser spray nozzles for aerial applications), addressing 

concerns of Chromobacterium subtsugae strain PRAA4-1T’s toxicity to honey bees, terrestrial 

arthropods, and aquatic invertebrates, has been added to the MBI-203 EP Bioinsecticide label. 
Chromobacterium subtsugae strain PRAA4-1T Page 17 of 36

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The Biopesticides and Pollution Prevention Division (BPPD) makes “no effect” determinations 

for direct effects, indirect effects, and affects to habitat (including designated critical habitat) for 

listed species for the following labeled applications of Chromobacterium subtsugae strain 

PRAA4-1T to the proposed crops:

• All foliar and soil directed liquid applications to the proposed crop plants treated within 

enclosed greenhouses

BPPD concludes that the following types of labeled applications of Chromobacterium subtsugae

strain PRAA4-1T to the proposed crops have the potential to cause direct effects, indirect effects, 

and effects to habitat (including designated critical habitat) to listed species. Further analyses are 

needed to characterize the effects that are likely to occur and the species potentially affected. 

• All outdoor foliar applications made by chemigation and aerial spray equipment.

V. ENVIRONMENTAL JUSTICE

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. EPA has this goal for all communities and persons 

across the United States. 

To help address potential environmental justice issues, during the 30-day public participation 

comment period, EPA sought information on any groups or segments of the population who, as a 

result their location, cultural practices, or other factors, may have atypical, unusually high 

exposure to Chromobacterium subtsugae strain PRAA4-1T

, compared to the general population.

No public comments were received on this particular matter.

For additional information regarding environmental justice issues, please visit EPA’s web site at 

http://www.epa.gov/compliance/environmentaljustice/index.html.
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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 Chromobacterium subtsugae strain PRAA4-1T pesticide products have

well-known properties. EPA has no knowledge that would contradict the claims made on the 

MBI-203 TGAI and MBI-203 EP Bioinsecticide labels, and such products are not expected to 

cause unreasonable adverse effects on the environment when used according to the label 

instructions. Criterion 2 is satisfied by the current product labels, as well as the data and 

information presented in this document. It is believed that the Chromobacterium subtsugae strain 

PRAA4-1T pesticide products will not cause any unreasonable adverse effects on the 

environment, and MBI-203 EP Bioinsecticide (end-use pesticide product), in particular, is likely 

to provide protection against insect and mite pests as claimed, satisfying criterion 3. Criterion 4 is

satisfied in that the Chromobacterium subtsugae strain PRAA4-1T pesticide products are not 

expected to cause unreasonable adverse effects when used according to label instructions. 

Therefore, MBI-203 TGAI and MBI-203 EP Bioinsecticide, containing Chromobacterium 

subtsugae strain PRAA4-1T as a new active ingredient, are eligible for registration under FIFRA 

section 3(c)(5) for the labeled uses. If the applicant proposes uses that are more extensive in the 

future (e.g., aquatic uses), EPA will likely require that additional data, in conjunction with a 

registration amendment request, be submitted.

VII. ACTIONS REQUIRED BY THE REGISTRANT

A. Final Printed Labeling

Before releasing pesticide products containing Chromobacterium subtsugae strain PRAA4-1T for 

shipment, the registrant is required to provide appropriate final printed labeling to EPA.

B. Terms of Registration

As terms of the MBI-203 EP Bioinsecticide registration, the registrant must submit the following 

data within one year of this product’s registration:

(1) Analysis of Samples (OCSPP Guideline 885.1400): A five-batch analysis, addressing all 

quality control parameters (i.e., dry weight, standard plate count, viscosity, density, and 

pH).
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(2) Storage Stability (OCSPP Guideline 830.6317) and Corrosion Characteristics (OCSPP 

Guideline 830.6320): The results of a two-year storage stability and corrosion 

characteristics study (ongoing). 

C. Reporting of Adverse Effects and Hypersensitivity Incidents

Notwithstanding the information stated in the previous sections, it should be clearly understood 

that certain, specific data are required to be reported to EPA as a requirement for maintaining the 

federal registration for a pesticide product. A brief summary of these types of data are described 

below.

Reports of all incidents of adverse effects to the environment must be submitted to 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).
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VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

ASAE American Society of Agricultural Engineers

BPPD Biopesticides and Pollution Prevention Division

BRAD Biopesticides Registration Action Document

CFR Code of Federal Regulations

cfu colony-forming unit(s)

cfu/kg colony-forming units per kilogram

cfu/mL colony-forming units per milliliter

cP centipoise

EDSP Endocrine Disruptor Screening Program

EP end-use product

EPA Environmental Protection Agency (the “Agency”)

FFDCA Federal Food, Drug, and Cosmetic Act

FIFRA Federal Insecticide, Fungicide, and Rodenticide Act

FQPA Food Quality Protection Act

FR Federal Register

g/mL gram per milliliter

LC50 median lethal concentration. A statistically derived concentration of a 

substance that can be expected to cause death in 50% of test animals. It is 

usually expressed as the weight of substance per weight or volume of water, 

air, or feed (e.g., mg/L, mg/kg, or ppm).

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

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

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

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

mg/kg milligrams per kilogram

mg/L milligrams per liter

MP manufacturing-use product

MRID No. Master Record Identification Number

NIOSH National Institute for Occupational Safety and Health

NRRL Northern Regional Research Laboratory

OCSPP Office of Chemical Safety and Pollution Prevention

OPP Office of Pesticide Programs

PC Code Pesticide Chemical Code

PP Pesticide Petition

PPE personal protective equipment

ppm parts per million

TGAI technical grade of the active ingredient

U.S. United States
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IX. BIBLIOGRAPHY

A. Studies Submitted to Support the Chromobacterium subtsugae strain PRAA4-1T

 Pesticide Product Registrations

479450-01 Koivunen M. 2009. Product Chemistry for MBI-203 TGAI. Unpublished study 

prepared by Marrone Bio Innovations, 190 pages.

479450-02 Koivunen M. 2009. Product Chemistry for MBI-203 EP. Unpublished study 

prepared by Marrone Bio Innovations, 143 pages.

479450-03 Oley S. 2009. MOI 203 Chromobacteria: Acute Oral Toxicity Up and Down 

Procedure in Rats. Project Number: 25936, P320/UDP. Unpublished study 

prepared by Eurofins/Product Safety Laboratories, 15 pages.

479450-04 Kuhn J. 2009. MOI 203 Chromobacteria: Acute Dermal Toxicity Study in Rats –

Final Report. Project Number: 12780/09. Unpublished study prepared by 

Stillmeadow, Incorporated, 11 pages.

479450-05 Younger C. 2009. MOI 203 Chromobacteria: Acute Inhalation Toxicity Study in 

Rats – Final Report. Project Number: 12781/09. Unpublished study prepared by 

Stillmeadow, Incorporated, 17 pages.

479450-06 Kuhn J. 2009. MOI 203 Chromobacteria: Acute Eye Irritation Study in Rabbits –

Final Report. Project Number: 12782/09. Unpublished study prepared by 

Stillmeadow, Incorporated, 16 pages.

479450-07 Kuhn J. 2009. MOI 203 Chromobacteria: Acute Dermal Irritation Study in 

Rabbits – Final Report. Project Number: 12783/09. Unpublished study prepared 

by Stillmeadow, Incorporated, 11 pages.

479450-08 Kuhn J. 2009. MOI 203 Chromobacteria: Skin Sensitization Study in Guinea Pigs 

– Final Report. Project Number: 12784/09. Unpublished study prepared by

Stillmeadow, Incorporated, 16 pages.

479450-09 Monds K. 2009. MOI 203 Chromobacteria: MPCA Mouse Safety Study – Final 

Report. Project Number: 12446/08. Unpublished study prepared by Stillmeadow, 

Incorporated, 10 pages.

479450-10 Monds K. 2009. MOI 203 Chromobacterium: Acute Injection 

Toxicity/Pathogenicity Pilot Study in Mice – Final Report. Project Number: 

12661/08. Unpublished study prepared by Stillmeadow, Incorporated, 11 pages.

479450-11 Monds K. 2009. MOI 203 Chromobacterium: Acute Injection Toxicity/Infectivity 

Study in Rats – Final Report. Project Number: 13056/09. Unpublished study 

prepared by Stillmeadow, Incorporated, 19 pages.
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479450-12 Gallagher S, Krueger H. 2009. MBI-203: A 48-Hour Static Acute Toxicity Test 

with the Cladoceran (Daphnia magna) – Final Report. Project Number: 

664A/103A. Unpublished study prepared by Wildlife International, Limited, 38 

pages.

479450-13 Hubbard P, Beavers J. 2009. MBI-203: An Avian Oral Pathogenicity and Toxicity 

Study in the Northern Bobwhite. Project Number: 664/101. Unpublished study 

prepared by Wildlife International, Limited, 50 pages.

479450-14 Gallagher S, Krueger H. 2009. MBI-203: A Five-Concentration Toxicity and 

Pathogenicity Test with the Fathead Minnow (Pimephales promelas) – Final 

Report. Project Number: 664A/102. Unpublished study prepared by Wildlife 

International, Limited, 61 pages.

479450-15 Morgan C. 2009. MBI-203: Nontarget Plants. Project Number: 4401/11, 1503/18, 

4501/16. Unpublished study prepared by Marrone Bio Innovations, 8 pages.

479450-16 Chanbusarakum L. 2009. MBI-203: Ladybird Beetle (Hippodamia convergens) 

Contact Assay. Project Number: 2702/145. Unpublished study prepared by 

Marrone Bio Innovations, 9 pages.

479450-17 Chanbusarakum L. 2009. MBI-203: Ladybird Beetle (Hippodamia convergens) 

Feeding Assay. Project Number: 104/47. Unpublished study prepared by Marrone 

Bio Innovations, 9 pages.

479450-18 Porch J, Krueger H. 2009. MBI-203: A Dietary Pathogenicity and Toxicity Study 

with Green Lacewing Larvae (Chrysoperla rufilabris) – Final Report. Project 

Number: 664/103B. Unpublished study prepared by Wildlife International, 

Limited, 46 pages. 

479450-19 Richards K. 2009. Evaluation of the Dietary Effect(s) of MBI 203 on Adult Honey 

Bees (Apis mellifera L.). Project Number: CAR/140/09. Unpublished study 

prepared by California Agricultural Research, Incorporated, 45 pages.

479450-20 Richards K. 2009. Evaluation of the Dietary Effect(s) of MBI 203 on Adult Honey 

Bees (Apis mellifera L.) – Final Report. Project Number: CAR/140/09B. 

Unpublished study prepared by California Agricultural Research, Incorporated, 12 

pages.

479450-21 Richards K. 2009. Evaluation of the Dietary Effect(s) of MBI 203 on Adult Honey 

Bees (Apis mellifera L.) – Final Report. Project Number: CAR/140/09A. 

Unpublished study prepared by California Agricultural Research, Incorporated, 11 

pages.

479450-22 Mileson B. 2009. Endangered Species Evaluation for MBI-203. Unpublished 

study prepared by Technology Sciences Group, Incorporated, 23 pages.
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479450-23 Mileson B. 2009. Response to Tier 1 Microbial Pesticide Data Requirements for 

MBI-203 TGAI. Unpublished study prepared by Technology Sciences Group, 

Incorporated, 29 pages.

479450-24 Mileson B. 2009. Response to Tier 1 Microbial Pesticide Data Requirements for 

MBI-203 EP. Unpublished study prepared by Technology Sciences Group, 

Incorporated, 77 pages.

479450-25 Pitts K. 2009. Published Literature to Support the Registration of MBI 203.

Unpublished study prepared by Marrone Bio Innovations, 275 pages.

481129-02 Koivunen M. 2010. Supplemental Product Identity Study, Secondary Metabolites 

(MBI-203). Unpublished study prepared by Marrone Bio Innovations, 136 pages.

481129-03 Chanbusarakum L. 2010. Supplemental Manufacturing Process Study, Cabbage 

Looper Potency Unit Justification (MBI-203). Unpublished study prepared by 

Marrone Bio Innovations, 25 pages. 

481129-04 Gallagher S, Krueger H. 2009. MBI-203: A 21-Day Life Cycle Toxicity and 

Pathogenicity Test with the Cladoceran (Daphnia magna). Project Number: 

664A/101. Unpublished study prepared by Wildlife International, Limited, 33

pages.

481129-05 Younger C. 2010. MBI 203: Parasitic Wasp (Aphidius matricariae), Nontarget 

Insect Microbial Testing – Final Report. Project Number: 13902/10. Unpublished 

study prepared by Stillmeadow, Incorporated, 16 pages. 

481129-06 Koivunen M. 2010. Supplemental Product Identity and Manufacturing Study, 

Replacement of Inert Ingredient for MBI-203 EP. Unpublished study prepared by 

Marrone Bio Innovations, 24 pages.

481129-07 Chanbusarakum L. 2010. Supplemental Manufacturing Process, Cabbage Looper 

Potency Unit Justification for MBI-203 EP. Unpublished study prepared by 

Marrone Bio Innovations, 27 pages.

481129-08 Pitts K. 2010. Response to Tier 1 Microbial Pesticide Data Requirements for 

MBI-203 EP: Supplemental Study for Revised EP Formulation. Unpublished 

study prepared by Marrone Bio Innovations, 123 pages.

481307-01 Chanbusarakum L. 2010. MBI 203 TGAI: Supplemental Product Identity Study, 

Mode of Action. Unpublished study prepared by Marrone Bio Innovations, 30 

pages.

484406-01 Pitts K. 2011. MBI 203 TGAI: 12-Month Storage Stability and Corrosion 

Characteristics Study. Unpublished study prepared by Marrone Bio Innovations, 

21 pages.
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484407-01 Pitts K. 2011. MBI 203 EP: 7-Month Storage Stability and Corrosion 

Characteristics Study. Unpublished study prepared by Marrone Bio Innovations, 

21 pages.

B. Environmental Protection Agency Risk Assessment Memoranda and Other Documents

U.S. EPA. 2011a. Chromobacterium subtsugae strain PRAA4-1T

. Memorandum from 

J.V. Gagliardi, Ph.D. and J.L. Kough, Ph.D. to J. Kausch dated April 21, 2011.

U.S. EPA. 2011b. Environmental Risk Assessment of Chromobacterium subtsugae strain 

PRAA4-1T (PC Code: 016329) for Section 3 Registrations of the Technical Product

(MBI-203 TGAI; EPA File Symbol 84059-O) and End Use Product (MBI-203 EP; EPA 

File Symbol 84059-RN) for Control of Insects and Mites in Ornamental Plants, Turf and 

Edible Crops, Memorandum from A. Gross and Z. Vaituzis, Ph.D. to J. Kausch dated 

July 7, 2011.

U.S. EPA. 2011c. Spray Drift Buffers for Chromobacterium subtsugae strain PRAA4-I. Analysis 

drafted by S. Borges and dated August 11, 2011.

U.S. EPA. 2011d. Spray Buffers Revised C. subtsugae. Spreadsheet assembled by S. Borges and 

dated August 17, 2011.

 

C. Other References

Aislabie J, Smith JJ, Fraser R, McLeod M. 2001. Leaching of bacterial indicators of faecal 

contamination through four New Zealand soils. Australian Journal of Soil Research

39:1397-1406.

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.

Lindow SE, Brandl MT. 2003. Microbiology of the phyllosphere. Applied and Environmental 

Microbiology 69:1875–1883.

Martin PAW, Gundersen-Rindal D, Blackburn M, Buyer J. 2007a. Chromobacterium subtsugae

sp. nov., a betaproteobacterium toxic to Colorado potato beetle and other insect pests. 

International Journal of Systematic and Evolutionary Microbiology 57:993–999.

Martin PAW, Hirose E, Aldrich JR. 2007b. Toxicity of Chromobacterium subtsugae to southern 

green stink bug (Heteroptera: Pentatomidae) and corn rootworm (Coleoptera: 

Chrysomelidae). Journal of Economic Entomology 100(3):680–684.

Pang L, McLeod M, Aislabie J, Šimůnek J, Close M, Hector R. 2008. Modeling transport of 

microbes in ten undisturbed soils under effluent irrigation. Vadose Zone Journal 7:97-

111.
Chromobacterium subtsugae strain PRAA4-1T Page 25 of 36

Biopesticides Registration Action Document

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.

U.S. EPA. 1996. Microbial Pesticide Test Guidelines – Background for Residue Analysis of 

Microbial Pest Control Agents (OPPTS 885.2000). Available from 

http://www.epa.gov/ocspp/pubs/frs/publications/Test_Guidelines/series885.htm.
Chromobacterium subtsugae strain PRAA4-1T Page 26 of 36

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APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS 

(40 CFR PART 158 – SUBPART V)

TABLE 1. Product Analysis Data Requirements for the Technical Grade of the Active Ingredient

(TGAI)/Manufacturing-Use Product (MP), MBI-203 TGAI (40 CFR § 158.2120)

OCSPP 

Guideline 

Number 

Data Requirement

Results

MRID No.

TGAI MP

885.1100 Product Identity Not applicable Submitted data fulfill the 

requirement for product 

identity. MBI-203 TGAI 

contains 100.00% by weight 

Chromobacterium subtsugae

strain PRAA4-1T cells and 

spent fermentation media 

(minimum of 1,500 Cabbage 

Looper Potency (or Killing) 

Units per milligram). 

479450-01

481129-02

481307-01

885.1200 Manufacturing Process Submitted data fulfill the requirement for manufacturing 

process.

479450-01

481129-03

Not applicable Deposition of a Sample 

in a Nationally 

Recognized Culture 

Collection

Chromobacterium subtsugae

strain PRAA4-1T is on 

deposit with the Agricultural 

Research Service Culture 

Collection (also known as the 

Northern Regional Research 

Laboratory Collection) in 

Peoria, Illinois under 

Accession Number NRRL B30655.

Not applicable 479450-01

885.1300 Discussion of Formation 

of Unintentional 

Ingredients

Submitted data fulfill the requirement for discussion of 

formation of unintentional ingredients.

479450-01

885.1400 Analysis of Samples Submitted data fulfill the requirement for analysis of samples. 479450-01

885.1500 Certification of Limits Not applicable Limits listed on the 

confidential statement of 

formula are 

adequate/acceptable.

479450-01

Additional Studies

830.1800 Enforcement Analytical 

Method

Not applicable Submitted data fulfill the 

requirement for an 

enforcement analytical 

method.

479450-01
Chromobacterium subtsugae strain PRAA4-1T Page 27 of 36

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TABLE 2. Product Analysis Data Requirements for the End-Use Product (EP), MBI-203 EP Bioinsecticide

(40 CFR § 158.2120)

OCSPP Guideline Number Data Requirement Results MRID No.

885.1100 Product Identity Submitted data fulfill the requirement for 

product identity. MBI-203 EP

Bioinsecticide contains 94.50% by weight 

Chromobacterium subtsugae strain 

PRAA4-1T cells and spent fermentation 

media (minimum of 1,000 Cabbage Looper 

Potency (or Killing) Units per milligram). 

479450-02

481129-06

885.1200 Manufacturing Process Submitted data fulfill the requirement for 

manufacturing process.

479450-02

481129-06

481129-07

Not applicable Deposition of a Sample in a 

Nationally Recognized Culture 

Collection

Not applicable Not applicable

885.1300 Discussion of Formation of

Unintentional Ingredients

Submitted data fulfill the requirement for 

discussion of formation of unintentional 

ingredients.

479450-02

885.1400 Analysis of Samples Submitted date fulfill the requirement for 

analysis of samples for purposes of FIFRA 

section 3(c)(5) registration. As a term of 

the MBI-203 EP Bioinsecticide

registration, EPA is requiring a fivebatch analysis, addressing all quality 

control parameters (i.e., dry weight, 

standard plate count, viscosity, density, 

and pH), be submitted within one year.

479450-02

885.1500 Certification of Limits Limits listed on the confidential statement 

of formula are adequate/acceptable.

479450-02

Additional Studies

830.1800 Enforcement Analytical 

Method

Submitted data fulfill the requirement for 

an enforcement analytical method.

479450-02
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TABLE 3. Physical and Chemical Characteristics for the Technical Grade of the Active Ingredient 

(TGAI)/Manufacturing-Use Product (MP), MBI-203 TGAI (40 CFR § 158.2120)

OCSPP 

Guideline 

Number 

Data Requirement

Results

MRID No.

TGAI MP

830.6302 Color Purple-blue Not applicable 479450-01

830.6303 Physical State Liquid Not applicable 479450-01

830.6304 Odor Grass-like Not applicable 479450-01

830.6313 Stability to Normal and 

Elevated

Temperatures, Metals,

and Metal Ions

Expected to be stable Not applicable 479450-01

830.6317 Storage Stability Stable for one year when stored per label directions. 479450-01

484406-01

830.6319 Miscibility Not applicable Not required because MBI-203 

TGAI is not an emulsifiable form 

of microbial pesticide (refer to test 

note #2 of 40 CFR § 158.2120(d)).

Not 

applicable

830.6320 Corrosion

Characteristics

Not applicable Not corrosive to packaging 

materials in a one-year study. 

479450-01

484406-01

830.7000 pH 4.1 (1% w/w) Not applicable 479450-01

830.7100 Viscosity Not applicable 5.4 cP 479450-01

830.7300 Density/Relative 

Density/Bulk Density 

(Specific Gravity)

1.01 g/mL Not applicable 479450-01
Chromobacterium subtsugae strain PRAA4-1T Page 29 of 36

Biopesticides Registration Action Document

TABLE 4. Physical and Chemical Characteristics for the End-Use Product (EP), MBI-203 EP Bioinsecticide

(40 CFR § 158.2120)

OCSPP Guideline Number Data Requirement Results MRID No.

830.6302* Color Purple-blue 479450-02

830.6303* Physical State Liquid 479450-02

830.6304* Odor Herbaceous 479450-02

830.6313 Stability to Normal and 

Elevated Temperatures, 

Metals, and Metal Ions

Not applicable Not applicable

830.6317 Storage Stability Expected to be stable for two years when 

stored per label instructions. As a term of 

the MBI-203 EP Bioinsecticide 

registration, EPA is requiring the 

results of a two-year storage stability 

and corrosion characteristics study 

(ongoing) be submitted within one 

year.

479450-02

484407-01

830.6319 Miscibility Miscible with water 479450-02

830.6320 Corrosion Characteristics Not expected to be corrosive to packaging 

materials. As a term of the MBI-203 EP

Bioinsecticide registration, EPA is 

requiring the results of a two-year 

storage stability and corrosion 

characteristics study (ongoing) be 

submitted within one year.

479450-02

484407-01

830.7000* pH 6.2 479450-02

830.7100 Viscosity 21.8 cP 479450-02

830.7300* Density/Relative Density/Bulk 

Density (Specific Gravity)

1.02 g/mL 479450-02

* According to 40 CFR § 158.2120, these data are only required for the technical grade of the active 

 ingredient. Since Marrone Bio Innovations included this information with its application for MBI-203 EP

 Bioinsecticide, it is summarized appropriately in this table.
Chromobacterium subtsugae strain PRAA4-1T Page 30 of 36

Biopesticides Registration Action Document

TABLE 5. Toxicology Data Requirements for the Technical Grade of the Active Ingredient (TGAI)/Manufacturing-Use 

Product (MP), MBI-203 TGAI (40 CFR § 158.2140)

OCSPP Guideline 

Number

Data Requirement Results MRID No.

TGAI MP

Tier I

885.3050 Acute Oral 

Toxicity/Pathogenicity

Waived based on the results of 

MRID Nos. 479450-11 and 

479450-03. 

Classification: Acceptable

Not applicable 479450-23

885.3150 Acute Pulmonary 

Toxicity/Pathogenicity

Waived based on the results of 

MRID Nos. 479450-11 and 

479450-05. 

Classification: Acceptable

Not applicable 479450-23

885.3200 Acute Injection 

Toxicity/Pathogenicity 

(Intravenous)

Not toxic, infective, and/or 

pathogenic to rats when dosed

intravenously at 3.1 x 106 cfu 

per animal.

Classification: Acceptable

Not applicable 479450-11

885.3400 Hypersensitivity 

Incidents

No hypersensitivity incidents, including immediate-type or 

delayed-type reactions of humans and domestic animals that 

occurred during research, development, or testing of the 

TGAI/MP, were reported by the applicant. Any future 

hypersensitivity incidents must be reported to EPA (refer to test 

note #3 of 40 CFR § 158.2140(d)). 

479450-23

885.3500 Cell Culture Not required because 

Chromobacterium subtsugae

strain PRAA4-1T is not a virus 

(refer to test note #4 of 40 

CFR § 158.2140(d)).

Not applicable Not 

applicable 

870.1100 Acute Oral Toxicity Not applicable Oral LD50 female rats > 5,000 

mg/kg

Classification: Acceptable

TOXICITY CATEGORY IV

479450-03

870.1200 Acute Dermal Toxicity Not applicable Dermal LD50 combined (male 

and female) rats > 5,050 mg/kg

Classification: Acceptable

TOXICITY CATEGORY IV

479450-04

870.1300 Acute Inhalation 

Toxicity

Not applicable Inhalation LC50 combined 

(male and female) rats > 2.12 

mg/L

Classification: Acceptable

TOXICITY CATEGORY IV

479450-05

870.2400 Acute Eye Irritation Not applicable MBI-203 TGAI was minimally 

irritating to the eyes of rabbits.

Classification: Acceptable

TOXICITY CATEGORY IV

479450-06

870.2500 Primary Dermal 

Irritation

Not applicable MBI-203 TGAI was slightly 

irritating to the skin of rabbits.

Classification: Acceptable

TOXICITY CATEGORY IV

479450-07

Tiers II and III

Not required for Chromobacterium subtsugae strain PRAA4-1T based on the lack of acute toxicity/pathogenicity in the Tier I 

studies.
Chromobacterium subtsugae strain PRAA4-1T Page 31 of 36

Biopesticides Registration Action Document

TABLE 5. Toxicology Data Requirements for the Technical Grade of the Active Ingredient (TGAI)/Manufacturing-Use 

Product (MP), MBI-203 TGAI (40 CFR § 158.2140)

OCSPP Guideline 

Number

Data Requirement Results MRID No.

TGAI MP

Additional Studies

870.2600 Skin Sensitization Not a dermal sensitizer

Classification: Acceptable

479450-08

Not applicable Safety Study 

(Subcutaneous

Injection)

Not toxic or pathogenic at > 106 cfu/mouse subcutaneously

Classification: Supplemental – Not a required study

479450-09

885.3200 Acute Injection 

Toxicity/Pathogenicity 

(Intravenous) Pilot 

Study

Provides no useful data as to the mechanism of toxicity observed

Classification: Supplemental – Not a required study

479450-10
Chromobacterium subtsugae strain PRAA4-1T Page 32 of 36

Biopesticides Registration Action Document

TABLE 6. Toxicology Data Requirements for the End-Use Product (EP), MBI-203 EP Bioinsecticide

(40 CFR § 158.2140)

OCSPP Guideline Number Data Requirement Results MRID No.

885.3050 Acute Oral 

Toxicity/Pathogenicity

Not applicable Not applicable

885.3150 Acute Pulmonary 

Toxicity/Pathogenicity

Not applicable Not applicable

885.3200 Acute Injection 

Toxicity/Pathogenicity

Not applicable Not applicable

885.3400 Hypersensitivity Incidents No hypersensitivity incidents, including 

immediate-type or delayed-type 

reactions of humans and domestic 

animals that occurred during research, 

development, or testing of the EP, were 

reported by the applicant. Any future 

hypersensitivity incidents must be 

reported to EPA (refer to test note #3 of 

40 CFR § 158.2140(d)).

Not applicable

885.3500 Cell Culture Not applicable Not applicable

870.1100 Acute Oral Toxicity Waived based on the results of MRID 

No. 479450-03 and because this 

formulation contains inert ingredients 

that are present in small amounts, are not 

expected to be of toxicological concern, 

and are cleared for food use.

Classification: Acceptable

TOXICITY CATEGORY III

479450-24

481129-08

870.1200 Acute Dermal Toxicity Waived based on the results of MRID 

No. 479450-04 and because this 

formulation contains inert ingredients 

that are present in small amounts, are not 

expected to be of toxicological concern, 

and are cleared for food use.

Classification: Acceptable

TOXICITY CATEGORY III

479450-24

481129-08

870.1300 Acute Inhalation Toxicity Waived based on the results of MRID 

No. 479450-05 and because this 

formulation contains inert ingredients 

that are present in small amounts, are not 

expected to be of toxicological concern, 

and are cleared for food use.

Classification: Acceptable

TOXICITY CATEGORY III

479450-24

481129-08

870.2400 Acute Eye Irritation Waived based on the results of MRID 

No. 479450-06 and because this 

formulation contains inert ingredients 

that are present in small amounts, are not 

expected to be of toxicological concern, 

and are cleared for food use.

Classification: Acceptable

TOXICITY CATEGORY III

479450-24

481129-08
Chromobacterium subtsugae strain PRAA4-1T Page 33 of 36

Biopesticides Registration Action Document

TABLE 6. Toxicology Data Requirements for the End-Use Product (EP), MBI-203 EP Bioinsecticide

(40 CFR § 158.2140)

OCSPP Guideline Number Data Requirement Results MRID No.

870.2500 Primary Dermal Irritation Waived based on the results of MRID 

No. 479450-07 and because this 

formulation contains inert ingredients 

that are present in small amounts, are not 

expected to be of toxicological concern, 

and are cleared for food use.

Classification: Acceptable

TOXICITY CATEGORY III

479450-24

481129-08

TABLE 7. Nontarget Organism Toxicity and Environmental Fate Data Requirements for the Technical Grade of the 

Active Ingredient (TGAI), Chromobacterium subtsugae strain PRAA4-1T (40 CFR § 158.2150)

OCSPP Guideline 

Number

Data Requirement Results MRID No.

Tier I

885.4050 Avian Oral Toxicity The no-observed-effect dosage of MBI-203 TGAI 

in this study was 4.0 x 1011 cfu/kg body 

weight/day for 5 days.

Classification: Acceptable

479450-13

885.4100 Avian Inhalation 

Toxicity/Pathogenicity

Data waiver rationale provides sufficient 

information to determine that pathogenicity to 

birds is not expected.

Classification: Acceptable

479450-23

885.4150 Wild Mammal 

Toxicity/Pathogenicity

Tests required by 40 CFR § 158.2140 are 

adequate and appropriate for assessment of 

hazards to wild mammals. Testing indicates no 

adverse effects to laboratory rats when dosed 

orally at 5,000 mg MBI-203 TGAI/kg body 

weight.

Classification: Acceptable

479450-03

885.4200 Freshwater Fish 

Toxicity/Pathogenicity

The study indicates no adverse effects to fathead 

minnow (Pimephales promelas) when dosed with 

Chromobacterium subtsugae at a rate of 8.24 x 

107 cfu/mL.

Classification: Acceptable

479450-14

885.4240 Freshwater Invertebrate 

Toxicity/Pathogenicity

Tests indicate toxicity to Daphnia magna, and the 

addition of aquatic uses would require further 

testing. 

Classification: Supplemental but acceptable 

for risk assessment

479450-12

481129-04

The following mitigating language is required in 

the Environmental Hazards section of the MBI203 EP Bioinsecticide label: “This product is 

toxic to aquatic invertebrates. Drift and runoff 

may be hazardous to aquatic organisms in water 

adjacent to treated areas.” Additionally, the MBI203 EP Bioinsecticide label will (1) bear a 

maximum application rate of 12 quarts of product 

per acre, (2) include a restriction that aerial 

applications not occur within 75 feet of aquatic 

habitats, (3) require aerial applicators to use 

medium or coarser spray nozzles, and (4) list 

certain requirements and instructions to further 

reduce potential for spray drift to offsite areas.
Chromobacterium subtsugae strain PRAA4-1T Page 34 of 36

Biopesticides Registration Action Document

TABLE 7. Nontarget Organism Toxicity and Environmental Fate Data Requirements for the Technical Grade of the 

Active Ingredient (TGAI), Chromobacterium subtsugae strain PRAA4-1T (40 CFR § 158.2150)

OCSPP Guideline 

Number

Data Requirement Results MRID No.

885.4280 Estuarine/Marine Fish and 

Invertebrate Testing

Not required because Chromobacterium 

subtsugae strain PRAA4-l

T will not be applied to 

water and is not expected to enter 

marine/estuarine environments in amounts that 

are significantly higher than naturally occurring 

concentrations (refer to test note #6 of 40 CFR § 

158.2150(e)).

Not applicable

885.4300 Nontarget Plant Testing Effects of mild phytotoxicity were observed in 

cucumbers at 5X and 10X the field application 

rates and in chrysanthemum flowers at all 

application rates. 

Classification: Supplemental but acceptable 

for risk assessment

479450-15

Even though effects in cucumbers and 

chrysanthemum flowers were mild, the MBI-203 

EP Bioinsecticide label will still require all users 

to test for phytotoxicity prior to pesticide 

application.

885.4340 Nontarget Insect Testing Tests indicate toxicity to ladybird beetles. There 

were no observed effects to green lacewing larvae 

or parasitic wasps. The dosing and duration of the 

parasitic wasp study, however, were insufficient 

for a conclusive determination of no effects. 

Classification: Supplemental but acceptable 

for risk assessment

479450-16

479450-17

479450-18

481129-05

The following mitigating language is required in 

the Environmental Hazards section of the MBI203 EP Bioinsecticide label: “This product is 

toxic to certain nontarget terrestrial arthropods. 

Minimize spray drift away from target area to 

reduce effects to nontarget insects.” Further 

testing would be required for the removal of this 

mitigating language from the MBI-203 EP

Bioinsecticide label. Additionally, the MBI-203 

EP Bioinsecticide label will list certain 

requirements and instructions to further reduce 

potential for spray drift to offsite areas. 

885.4380 Honey Bee Testing Tests indicate toxicity to Apis mellifera. 

Classification: Supplemental but acceptable 

for risk assessment

479450-19

479450-20

479450-21

The following mitigating language is required in 

the Environmental Hazards section of the MBI203 EP Bioinsecticide label: “This product is 

highly toxic to bees exposed to direct treatment or 

residues on blooming crops or weeds. Do not 

apply this product or allow it to drift to blooming 

crops or weeds if bees are visiting the treatment 

area.” Further testing would be required for the 

removal of this mitigating language from the 

MBI-203 EP Bioinsecticide label. Additionally, 

the MBI-203 EP Bioinsecticide label will list 

certain requirements and instructions to further 

reduce potential for spray drift to offsite areas.
Chromobacterium subtsugae strain PRAA4-1T Page 35 of 36

Biopesticides Registration Action Document

TABLE 7. Nontarget Organism Toxicity and Environmental Fate Data Requirements for the Technical Grade of the 

Active Ingredient (TGAI), Chromobacterium subtsugae strain PRAA4-1T (40 CFR § 158.2150)

OCSPP Guideline 

Number

Data Requirement Results MRID No.

Tiers II, III, and IV

Not required for Chromobacterium subtsugae strain PRAA4-1T based on the current uses and application methods.

Additional Studies

Not applicable Endangered Species 

Evaluation

Sufficient information was provided to determine 

that any potential risk from use of MBI-203 

TGAI to endangered mammalian, avian, or fish 

species would likely be below the level of 

concern, and that adverse effects to endangered 

plant species are unlikely. Sufficient information 

was not provided to determine the potential risk 

to endangered nontarget insects or aquatic 

invertebrates. BPPD will make an independent 

endangered species determination.

Classification: Supplemental – not a required 

study

479450-22
Chromobacterium subtsugae strain PRAA4-1T Page 36 of 36

Biopesticides Registration Action Document

APPENDIX B. PESTICIDE PRODUCTS

EPA 

Registration 

Number

Registration 

Name

Percentage 

Active 

Ingredient

Formulation 

Type

Use Site(s) Method(s) 

of 

Application

Application 

Rate

Target 

Pest

84059-9 MBI-203 

TGAI

100.00% Technical N/A N/A N/A N/A

84059-10 MBI-203 EP

Bioinsecticide

94.50% End Use –

Liquid

Various 

agricultural

and 

greenhouse 

crops (e.g., 

vegetables, 

ornamentals, 

and turf)

Standard 

ground or 

aerial spray 

application 

equipment 

and 

chemigation

2–12 quarts 

per acre

Various 

insects and 

mites