BIOPESTICIDES REGISTRATION ACTION DOCUMENT



## Chromobacterium subtsugae strain PRAA4

- 1 T



### Pesticide Chemical (PC) Code: 016329

U.S. Environmental Protection Agency Office of Pesticide Programs Biopest icides and Pollution Prevention Division

Version # 4

J u ly 2 1 , 2011Chromobacterium subtsugae strain PRAA4

- 1 T Page 2 of 34 Biopesticides Registration Action Document

TABLE OF CONTENTS

I. EXECUTIVE SUMMARY

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

II. ACTIVE INGREDIENT OVERVIEW

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

III. REGULATORY BACKGROUN D

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

A. Applications for Pesticide Registration

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

B. Food Tolerance Exemption

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

IV. RISK ASSESSMENT

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

A. Produ ct Analysis Assessment (40 CFR § 158.2120)

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

B. Human Health Assessment (40 CFR § 158.2140)

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

C. Environmental A ssessment (40 CFR § 158.2150)

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

V. ENVIRONMENTAL JUSTIC E

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

VI. RISK MANAGEMENT DECI SION

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

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

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

VIII. GLOSSARY OF ACRONYMS AND ABBREVI ATIONS

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

IX. BIBLIOGRAPHY

................................ ................................ ................................ ........... 20

A. Studies Submitted to Support the Chromobacterium subtsugae strain PRAA4

- 1 T

Pesticide Product Registrations

................................ ................................ ................................ ..... 20

B. Environmental Protection Agency Risk Assessment Memoranda

................................ ............. 23

C. Other References

................................ ................................ ................................ ............................. 23

APPENDIX A. MICROBI AL PESTICIDES DATA R EQUIREMENTS

............................ 24

APPENDIX B. PESTICIDE PRODUCTS

................................ ................................ ............... 34Chromobacterium subtsugae strain PRAA4

- 1 T Page 3 of 34 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 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

Regulations

Sheryl K. Reilly, Ph.D. Chief, Microbi al Pesticides Branch

Jeannine Kausch Regulatory Action LeaderChromobacterium subtsugae strain PRAA4

- 1 T Page 4 of 34 Biopesticides Registration Action Document

I. EXECUTIVE SUMMARY

Chr omobacterium subtsugae strain PRAA4

- 1 T is a gram

- negative, violet

- pigmented bacterium

that was isolated from soil under a n eastern hemlock ( Tsuga c anadensis ) in the C atoctin 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 ) lar vae, 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

- 1 T

- treated

diet resulted in reduc ed 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 inhi bition of feeding. Given the insecticidal capabilities of Chromobacterium subtsugae strain PRAA4

- 1 T , Marrone Bio Innovations has proposed to register both a manufacturing

- use pesticide product, MBI

- 203 TGAI (EPA File Symbol 84059

- O), and an end

- use pestici de product, MBI

- 203 EP (EPA File Symbol 84059

- RN), containing

thi s bacterium. MBI

- 203 EP is proposed for use to control label

- specified insect and mite p ests (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 have 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

- 1 T pesticide products. Overall, such

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

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

.

P roduct analysis data requirements for Chromobacterium subtsugae strain PRAA4

- 1 T , including product chemi stry and composition, analysis of samples, and physical and chemical

characteri stics, were fulfilled by acceptable guideline studies.

Ade quate mamm alian toxicology data and information were submitted to support the

Chromobacterium subtsugae strain PRAA4

- 1 T pesticide products. The acute injection toxicity/pathogenicity study showed t hat, at a single high dose, Chromobacterium subtsugae

strain PRAA4

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

. Acute p ulmonary and oral toxicity/patho genicity tests were waived after considering the results

of the intravenous test and MBI

- 203 TGAI

- specific toxicity tests ( acute or al and inhalation)

. Moreover, no hypersensitivity incidents, occurring during research, development, or testin g of Chromobacterium subtsugae strain PRAA4

- 1 T , 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 hyper sensitivity incidents, testing at higher tiers (i.e., Tiers II and III) was not required.

EPA has made a preliminary conclusion 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

- 1 T . No dietary risks are expected from

use of Chromobacterium subtsugae strain PRAA4

- 1 T as a n active ingredient in pesticideChromobacterium subtsugae strain PRAA4

- 1 T Page 5 of 34 Biopesticides Registration Action Document

products. Significant dietary exposure

— through food a nd drinking water

— is not anticipated given the biological nature of this bacterium to degrade due to predation by other biological organisms and exposure to environmental factors (e.g., sunlight and varying temperatures ) after application

. I f oral exposure should occur through food or drinking water, such exposure to Chromobacterium subtsugae strain PRAA4

- 1 T is expected to be several orders of magnitude

lower than the dose s used in both the acute or al toxicity and acute injection toxicity/pathogenicity

test s , during which no toxic , infective , and/or pathogenic effects were observed in rats . Finally

, nonoccupational exposure is considered unlikely for Chromobacterium subtsugae strain

PRAA4

- 1 T as all currently proposed uses occur in distinctly agricultu ral se ttings; the re are no proposed uses for residential areas.

Despite the low toxicological profile of Chromobacterium subtsugae strain PRAA4

- 1 T , baseline personal protective equipment (PPE) is required for handlers that may be exposed to the active ingredien t, due to their occupation, for prolonged periods or numerous times . Handlers working

with Chromobacterium subtsugae strain PRAA4

- 1 T 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 N

95, R

- 95, or P

- 95. EPA may require additional PPE, other than the standard described above, on a product

- specific basis

.

The data a nd data waiver rationale, submitted by the applicant to support the pesticide products containing Chromobacterium subtsugae strain PRAA4

- 1 T , 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 has performed an environmental risk assessment based on the data and data waiver rationale provided by the applicant and has determined that the proposed

use of Chromobacterium subtsugae strain PRAA4

- 1 T does not pose significant risk to nontarget organisms when used according to label directions. Based on study results, there are concerns of toxicity to terres trial 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” det erminations for direct effects, indirect effects

, and effects to habitat (including designated critical habitat) to listed species for all foliar and soil

directed applications to crop pl ants treated within greenhouses. On the other hand, EPA

concludes tha t 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 an nounced a new policy to provide a more meaningful opportunity for the public to participate i n 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 appli cations: new active ingredients; first food uses; first outdoor uses; first residential use s; 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 proposed

pe sticide products containing Chromobacterium subtsugae strain PRAA4

- 1 T

, a new active ingredient, are subject to a 30

- day comment period. In addition to containing a new active ingre dient, the registration of these products would result in both the first outdoor use and firstChromobacterium subtsugae strain PRAA4

- 1 T Page 6 of 34 Biopesticides Registration Action Document

food use for Chromobacterium subtsugae strain PRAA4

- 1 T

. 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

- 2010

- 0058. The fol lowing documents are available for comment in EPA

- HQ

OPP

- 2010

- 0058: (1) draft Chromobacterium subtsugae strain PRAA4

- 1 T Biopesticides Registration Action Document (BRAD) ; (2) environmental risk assessment for

Chromobacterium subtsugae strain PRAA4

- 1 T ; (3) draft produ ct label for the manufacturing

- use product, MBI

- 203 TGAI (EPA File Symbol 84059

- O); and (4) draft product label for the end

- use product, MBI

- 203 EP (EPA File Symbol 84059

- RN). While a final decision on registration is contingent upon review and consideration of public comments, EPA presently believes t hat, based upon the risk assessment and informat ion submitted in support of MBI

- 203 TGAI and

MBI

- 203 EP, it is in the best interest of the public and the environment to issue these registrations. The basis for this preliminary decision can be found in the risk assessment for Chromobacterium subtsugae strain PRAA4

- 1 T , which is characterized throughout this BRAD.

II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Chromobacterium subtsugae strain PRAA4

- 1 T

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 Pest icide: Microbial Pesticide

– Insecticide

See Appendix B for specific information (e.g., use sites, application rate s , methods of application, formulation type s , and target pests) regarding the registered pesticide p roducts containing this active ingredient.

III. REGULATORY BACKGROUND

A. Applications for Pesticide Registration

On December 22, 2009

, Marrone Bio Innovations (address: 2121 Second Street

, Suite B

- 107; Davis, California 95618 ) submitted application s to register a manufacturing

- use product, MBI

203 TGAI (EPA File Symbol 84059

- O )

, and an end

- use product, MBI

- 203 EP (EPA File Symbol

84059

- RN)

, under FIFRA section 3. On M arch 10, 2010

, EPA announced receipt of these application s to register pesticide pro duct s containing a new active ingredient ( 75 Federal Register (FR) 11175 ) and opened a 3 0

- day public comment period pursuant to the provisions of FIFRA section 3(c)(4). No commen ts were received following this publication.Chromobacterium subtsugae strain PRAA4

- 1 T Page 7 of 34 Biopesticides Registration Action Document

B. Food Tolerance Exemption

Concurrent with its registration application s 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

- 1 T
(Pesticide Petition (PP) 9F7674 )

. In the Federal Register of March 10, 2010 ( 75 FR 1117 1 ), EPA

announced that Marrone Bio Innovations Incorporated proposed to establish an exemption from the requirement of a tolerance for residues of the insecticide

, Chromobacterium subtsugae strain

PRAA4

- 1 T

, in or on all food commodities and opened a 30

- da y comment period. No comments were received following this publication.

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 th ese 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 ” indicat es 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 acc eptable (“ 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 re viewer will

often state that t he study is not required by 40 C FR P art 158. Both “ acceptable” and “supplemental ” studies may be used in the risk assessment process as appropriate. An “unacceptable ” rating indicates that new data need to be submitted.

For t he acu te toxicity data requirements, Toxicity C ategories are assigned based on the hazard(s) identified from studies and/or other infor mation submitted to EPA in support of a pesticide registration. The active ingredient or particular product is classifie d into Toxicity Category I, II, III, or IV, where Toxicity Category I indicates the highest toxicity and Toxicity Category IV indicates the lowest toxicity.

A. Product Analysis Assessment ( 40 CFR § 158.2120 )

A ll product analysis data requirements for Chromobacterium subtsugae strain PRAA4

- 1 T 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.Chromobacterium subtsugae strain PRAA4

- 1 T Page 8 of 34 Biopesticides Registration Action Document

B. Human Health Assessment ( 40 CFR § 158.2140 )

1. Toxicity

All toxicology data requirements for Chromobacterium subtsugae strain PRAA4

- 1 T have been

fulfilled

. Acceptable Tier I mammalian toxicolo gy data and information support registration of

the Chromobacterium subtsugae strain PRAA4

- 1 T pesticide products

. Furthermore, Tier II and Tier III studies were not required for Chromobacterium subtsugae strain PRAA4

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

For a compr ehensive 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 (O ffice of Chemical Safety and Pollution Prevention (O CSPP ) Guideline 885.3050; Master Record Identification Number ( MRID No. ) 479450

- 23 ) : Chromobacterium subtsugae occurs naturally in soil. A literature search (Agricola, Toxline, and U.S. National Library of Medicine PubMed) did not identify any reports of human or animal infection by Chromobacterium subtsugae

. In an acute up and down oral toxicity study (MRID

No. 479450

- 03), three female rats were gav aged 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, adver se effects, or abnormal behavior. The acute oral median lethal dose (LD 50 ) 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 mat erial ( 2.62 x 10 8 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 necrop sy 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 node s, lungs, brain, liver, spleen, and cecum contents by day 3. The test material was nontoxic and noninfective. These results suggest that the test material would not be infective by other routes. Sufficient information was supplied and deemed acceptable . Th us, the requirement for acute oral toxicity/pathogenicity testing was waived

.

Acute Pulmonary Toxicity/Pathogenicity (OCSPP Gui deline 885.3150; MRID No. 479450

- 23) : Chromobacterium subtsugae occurs naturally in soil. A literature search (Agricola, Toxline , and U.S. National Library of Medicine PubMed) did not identify any reports of human or animal infection by Chromobacterium subtsugae . 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 (LC 50 ) 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 10 8 cfu/mL). Three males and three females from the active test material group were sacrificed and necropsied on days 0, 3, 7, 14,Chromobacterium subtsugae strain PRAA4

- 1 T Page 9 of 34 Biopesticides Registration Action Document

and 21 after treatment. Tissue and blood samples were collected at necropsy and cultured to provide qu alitative 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, splee n, and cecum contents by day 3. The test material was nontoxic and noninfective. These results suggest that the test material would not be infective by other routes. Sufficient information was supplied and

deemed acceptable . Thus, the requirement for acute pulmonary toxicity/ pathogenicity testing was

waived

.

Acute Injection Toxicity/ Pathogenicity (Intravenous)

– Rat (OCSPP Guidel ine 885.3200; MRID

No. 479450

- 11) : In an acute intravenous injection toxicity and pathogenicity study, groups of young adult Spra gue

- Dawley rats were injected at 3.1 x 10 6 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 10 6 cfu per animal

. This study was rated acceptable

.

Hypersensitivity Incidents (OCSPP Guideline 885.3400; MRID No. 47 9450

- 23 ) : No hypersensitivit y 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

- 1 T

, were reported by the applicant . Any future hypersensitivity incidents must be reported per OCSPP Guideline 885.3400.

Cell Culture (OCSPP Guideline 885.3500) : This study is not required because Chromobacterium subtsugae strain PRAA4

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

b. Acute Toxi cology 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 subtsug ae strain PRAA4

- 1 T
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 Adm inistrator 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, androg en, or thyroid (E, A, or T) hormonalChromobacterium subtsugae strain PRAA4

- 1 T Page 10 of 34 Biopesticides Registration Action Document

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 wh ere 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 a nd 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 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

- 1 T is not amon g 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 pesti cide chemicals. Accordingly, EPA anticipates issuing future EDSP

orders/data call

- ins for all pesticide a ctive 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://w ww.epa.gov/endo/

.

2

. Food Quality Protection Act (FQPA) 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 resi due 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 dieta ry 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 pestic ide 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 aggreg ate exposure t o pesticide residues. First, EPA determines the tox icity 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 s ection 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 thi s information to human risk. EPA has also co nsidered available information concerning the variability of the sensitivities of major identifiable subgroups of consumers,Chromobacterium subtsugae strain PRAA4

- 1 T Page 11 of 34 Biopesticides Registration Action Document

including infants and children.

Based on the acute toxicity/pathogenicity data and information discussed previously and presented i n Table 5 in Appendix A , t he data required for a FQPA risk assessment for Chromobacterium subtsugae strain PRAA4

- 1 T 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 bui ldings (residential and other indoor uses).

Dietary Exposure and Risk Characterization : Dietary exposure to the naturally occurring soil bacterium, Chromobacterium subtsugae strain PRAA4

- 1 T , although a possibility, is anticipated to be negligible. Due to the biological nature of Chromobacterium subtsugae strain PRAA4

- 1 T
and its propensity for thriving in soil, it is not expected to persist or accumulate in any reservoirs on plants or food commodities. Rather, after application, it will naturally degrade du e to predation by other biological organisms and exposure to environmental factors (e.g., sunlight and varying temperatures )

. If o ral exposure should occur through food, such exposure to Chromobacterium subtsugae strain PRAA4

- 1 T is expected to be several o rders of magnitude

lower than the dose s used in both the acute or al toxicity and acute injection toxicity/pathogenicity

tests, during which no toxic , infective, and/or pathogenic effects were observed in rats.

Drinking Water Risk Characterization : Exposur e of humans to residues of Chromobacterium subtsugae strain PRAA4

- 1 T in consumed drinking water is unli kely. The proposed use patterns, use sites, and application methods for Chromobacterium subtsugae strain PRAA4

- 1 T do not include direct application to aq uatic environments , thereby limiting contact with surface water

. Furthermore, ground water is not expected to have significant exposure to Chromobacterium subtsugae strain PRAA4

- 1 T since the organism would likely be filtered out by the particulate

nature o f soils and is not known to be able to survive in water or deep soil. In the event that Chromobacterium subtsugae strain PRAA4

- 1 T is transferred to surface or ground water intended for eventual human consumption, the bacterium would not survive the conditi ons water is subjected to in a drinking water treatment facility, including high temperatures, chlorination, pH adjustments, and/or filtration. Even if oral exposure should occur through consumed drinking

water, EPA concludes that there is a reasonable cer tainty th at no harm will result from exposure

to such residues because of the lack of toxicity, infectivity, and or pathogenicity in the previously described toxicological studies (see section IV(B)(1)(a) and Table 5 in Append ix A )

.

Nonoccupational, Residential Risk Characterization : Nonoccupational exposure is not expected

for Chromobacterium subtsugae strain PRAA4

- 1 T as all proposed uses occur in distinctly agricultural settings, and there are no proposed uses for residenti al areas.Chromobacterium subtsugae strain PRAA4

- 1 T Page 12 of 34 Biopesticides Registration Action Document

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 informati on 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

- 1 T to share a common mechanism of toxicity with any other s ubstances, and Chromobacterium subtsugae strain

PRAA4

- 1 T does not appear to produce a toxic metabolite as its mode of action against the target
pest s . For the purposes of this tolerance action, therefore, EPA has assumed that Chromobacterium subtsugae stra in PRAA4

- 1 T does not have a common mechanism of toxicity

with other substances. For information re garding EPA ’s efforts to determine which chemicals have a common mechanism of toxicity and to evaluate the cumulative ef fects o f such chemicals,

see EPA ’s web site at http://www.epa.gov/pesticides/cumulative

.

c. Determination of Safety for the U.S. Population, Infants and Children

FFDCA sectio n 408(b)(2)(C) provides that EPA shall assess the availabl e 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 mechan ism of toxicity. In addition, FFDCA sectio n 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 complet enes s 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 FQPA Safety Factor. In applying this provision, EPA ei ther 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 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

- 1 T . This includes all anticipated dietary exposures and all other exposures for which there is reliable information. EPA has arrived at this conclusion

because the data and information available on Chromobacterium subtsugae strain PRAA4

- 1 T do

not dem onstrate toxic, pathogenic, and/ or infective potential to mammals. Thus, there are no threshold effects of concern and, as a result, the provision requiring an additional margin of safety does not apply.

3. Occupational Exposure and Risk Characterizatio n

Handler exposure to Chromobacterium subtsugae strain PRAA4

- 1 T is not expected to pose any undue risk. Regardless, appropriate personal protective equipment and precautionary statements are required on pesticide product label s to mitigate any potential r isks to pesticide handlers due to prolonged or numerous exposure s . Handlers applying Chromobacterium subtsugae strain

PRAA4

- 1 T end

- use 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 ofChromobacterium subtsugae strain PRAA4

- 1 T Page 13 of 34 Biopesticides Registration Action Document

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

- 1 T in light of the

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

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

a nd children in particu lar, will result when Chromobacterium subtsugae strain PRAA4

- 1 T
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 PRAA 4

- 1 T

, 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 m ethods. EPA has performed an environmental risk assessment based on the data and data waiver rationale provided by the applicant and has determined that the proposed use of Chromobacterium subtsugae strain PRAA4

- 1 T does not pose significant risk to nontarg et 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 compr ehensive 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. Terrestr ial Animals and Plants

The proposed 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 a nd, 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 MBI

203 TGAI (active ingredient, 100.00% Chromobacterium subtsugae strain PRAA4

- 1 T cells and spent fermentation media) to northern b obwhite ( Colinus virgini anus ). Fourteen

- day

- old chicks were dosed with the test material (4.0 x 10 1 1 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

additiona l 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

- 1 T

.Chromobacterium subtsugae strain PRAA4

- 1 T Page 14 of 34 Biopesticides Registration Action Document

In a n acute oral toxicity study (MRID No. 479450

- 03 )

, three fasted young adult female Sprague

Dawley 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 ro o m 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 n ecropsy

. This study demonstrates that toxic ity/pathogenicity to wild mammals is not expected from labeled applications of C hromobacterium subtsugae strain

PRAA4

- 1 T

.

Nontarget Insects (OCSPP Guideline 885.4340) and Honey Bees (OCSPP Guideline 885.4380) : Nontarget insect studies were submitted for l adybird 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 th e 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) was

submitted that fulfille d testing requirements; there were no observed effects from tes t
concentrations of Chromobacterium subtsugae strain PRAA4

- 1 T , and the LC 50 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 purpos es 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

- 1 T to which the wasps were exposed was not adequately characterized and the du ration 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

- 1 T (MRID No s

. 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

- 1 T . To address

these concerns and to reduce exposu re to nontarget insects and honey bees that may occur through outdoor ground and aerial applications , EPA is requiring aerial drift reduction information, as well as the following mitigation language

, on the MBI

- 203 EP label :

Environmental Hazards Statements

This product is highly toxic to bees expos ed 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.

This product is toxic to certain nontarget terrestrial arthropods. Minimize s pray drift away from target area to reduce effects to nontarget insects.

Nontarget Plants (OCSPP G uideline 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

- 1 T occurs naturally

in soils and i s 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,Chromobacterium subtsugae strain PRAA4

- 1 T Page 15 of 34 Biopesticides Registration Action Document

the MBI

- 203 EP 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 E stuarine/Marine Fish and Invertebrates (OCSPP Guideline 885.4280) : MBI

203 EP (94.50% Chromobacterium subtsugae strain PRAA4

- 1 T ) is not labeled for aquatic uses

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

de monstrated 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 rat ionale for testing on estuarine/marine organisms. The freshwater fish study (MRID No. 479450

- 14 ) fulfill s the testing requirements and did not show any adverse effects to fathead minnows ( Pimephales promelas )

. Toxicity was shown in two supplemental studies (MRID No s

. 479450

- 12 and 481129

- 04)

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

PRAA4

- 1 T 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 is not intended for direct application into the estuarine or marine environment or expected to enter such environment in significant c oncentrations.

Even though MBI

- 203 EP is not applied directly to aquatic environments, EPA is requiring specific a erial drift re duction t ext , as well the following mitigation language

, to reduce the potential for exposure to aquatic i nvertebrates :

Environmental Hazards

This product is toxic to aquatic invertebrates. D rift and runoff may be hazardous to aquatic organisms in water adjacent to treated areas

.

Buffer Zone for Aerial Applications (in the Directions for Use)

Do not apply within 150 feet of aquatic habitats (such as, but not limited to, lakes, reservoirs, rivers, streams, marshes, ponds, estuaries, and commercial fish ponds).

c. Open Literature Information

Literature searches were performed to determine if other effec ts 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 Wa ternet 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

– Ag ricultural Research Service concerning the toxicity of Chromobacterium subtsugae toChromobacterium subtsugae strain PRAA4

- 1 T Page 16 of 34 Biopesticides Registration Action Document

Colorado p otato b eetle and other insect pests (Martin et al . 2007a) , as well as to s outhern g reen

s tink b ug and c orn r ootwor m (Martin et al . 2007b) . Neither document provid ed additional information on environmental or nontarget effects useful for the purposes of this risk assessment.

2. Environmental Fate Data

As t he information provided , for the current uses and application methods, is sufficient to both satisfy the Ti er I nontarget organism data requirements and for nontarget organism risk

asse ssment for Chromobacterium subtsugae strain PRAA4

- 1 T , f urther testing at higher tier levels (i.e., Tiers II, III, and IV) is not required.

3

. Threatened and Endangered Spe cies 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 stra in PRAA4

- 1 T . 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 wer e 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 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 invertebra tes and other aquatic organisms. Mitigation language (e.g., requirement for a buffer be tween aquatic and treatment areas for aerial applications), addressing concerns of Chromobacterium subtsugae strain PRAA4

- 1 T ’s toxicity to honey bees, terrestrial arthropods , and aquatic invertebrates, has been added to the MBI

- 203 EP

label

.

The Biopest icides 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 Chromobacteri um subtsugae strain

PRAA4

- 1 T to the proposed crops:

**•** All foliar and soil directed liquid ap plications to the proposed crop plants trea ted within enclosed greenhouses

BPPD concludes that the following types of labeled application s of Chromobacterium subtsu gae strain PRAA4

- 1 T 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 likel y to occur and the species potentially affected.

**•** All outdoor foliar applications made by chemigation and aerial spray equipment.Chromobacterium subtsugae strain PRAA4

- 1 T Page 17 of 34 Biopesticides Registration Action Document

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 soci oeconomic 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. Mea ningful 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, EPA seeks information on any groups or segments of the population who, as a result their location, cultural practices, or other factors, may

have at ypical, unusually high exposure to Chromobacterium subtsugae strain PRAA4

- 1 T

, compared to the general population. Please comment if you are aware of any subpopulations that may have atypical, unusually high exposure compared to the general population.

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

.

V I

. RISK MANAGEM ENT 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 require d 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 practi ce, it will not generally cause unreasonable adverse effects on the

environment.

To satisfy criterion 1, the Chromobacterium subtsugae strain PRAA4

- 1 T pesticide products have

well

- known properties. EPA has no knowledge that would contradict the cla ims made on the

MBI

- 203 TGAI and MBI

- 203 EP label s, 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 label s, as well a s the data and information presented

in this document. It is believed that the Chromobacterium subtsugae strain PRAA4

- 1 T pesticide products will not cause any unreasonable adverse effects on the environment, and MBI

- 203 EP (end

- use product) , in particular, is likely to provide protection against insect and mite pests asChromobacterium subtsugae strain PRAA4

- 1 T Page 18 of 34 Biopesticides Registration Action Document

claimed, satisfying criterion 3. Criterion 4 is satisfied in that the Chromobacterium subtsugae

strain PRAA4

- 1 T pesticide products are not expected to cause unreasonable adverse effects when used according to label instructions. Therefore, MBI

- 203 TGAI and MBI

- 203 EP, containing Chromobacterium subtsugae strain PRAA4

- 1 T 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

.

V I I

. ACTIONS REQUIRED BY THE REGISTRANT

A. Final Printed Label ing

Before releasing pesticide products containing Chromobacterium subtsugae strain PRAA4

- 1 T 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 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, viscos ity, density, and pH). (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 pest icide 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 hyperse nsitivity (including both suspected and confirmed incidents) must be reported to EPA under the provisions of 40 CFR § 158.2140(d).Chromobacterium subtsugae strain PRAA4

- 1 T Page 19 of 34 Biopesticides Registration Action Document

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

c fu colony

- forming unit (s) cfu/kg colony

- forming units per kilogram

cfu/mL colony

- forming unit s per milliliter

cP centipoise

EDSP Endocrine Disru ptor 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 gra m per milliliter

LC 50 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).

LD 50 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 milligram s per kilogram mg/L milligram s 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 techn ical grade of the active ingredient

ULV ultra low volumeChromobacterium subtsugae strain PRAA4

- 1 T Page 20 of 34 Biopesticides Registration Action Document

I X. BIBLIOGRAPHY

A. Studies Submitted to Support the Chromobacterium subtsugae strain PRAA4

- 1 T

Pesticide Product Registrations

479450

- 01 Koivunen M. 2009

. Product Chemistry for MB I

- 203 TGAI . Unpublished study

pr epared by Marrone Bio Innovations

, 190 p ages

.

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 p ages

.

479450

- 04 Kuhn J

. 2009

. MOI 203 Chromobacteria : Acute Dermal Toxicity Study in Rats

– Fina l Report. Project Number: 12780/09 . Unpublished study prepared by Stillmeadow, Incorporated

, 11 p ages

.

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 p ages

.

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 p ages

.

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 p ages

.

479450

- 08 Kuhn J. 2009

. MOI 203 Chromobacteria: Skin Sensitizati on Study in Guinea Pigs

– Final Report. Project Number: 12784/09 . Unpublished study prepared by Stillmeadow, Incorporated, 16 p ages

.

479450

- 09 Monds K

. 2009

. MOI 203 Chromobacteria: MPCA Mouse Safety Study

– Final Report. Project Number: 12446/08 . Unpubli shed study prepared by Stillmeadow, Incorporated

, 10 p ages

.

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 p ages

.

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 p ages

.Chromobacterium subtsugae strain PRAA4

- 1 T Page 21 of 34 Biopesticides Registration Action Document

479450

- 12 Gallaghe r 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

p ages

.

479450

- 13 Hubbard P, Beavers J. 20 09. 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 p ages

.

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 p ages

.

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 p ages

.

479450

- 16 Chanbusarakum L

. 2009. MBI

- 203: Ladybird Beetle ( Hippodamia convergens ) Contact Assay. Project Number: 2702/145 . Unpublished study prepared by

Ma rrone Bio Innovations, 9 p ages

.

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

N umber: 664/103B . Unpublished study prepared by Wildlife International, Limited, 46 p ages

.

479450

- 19 Richards K. 2009. Evalu ation 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 p ages

.

479450

- 20 Richards K. 2009. Evaluation of the Dietary E ffect(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 p ages

.Chromobacterium subtsugae strain PRAA4

- 1 T Page 22 of 34 Biopesticides Registration Action Document

479450

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

MBI

- 203 TGAI. Unpublished study prepared by Technology Sciences Group, Incorporate d, 29 p ages

.

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 th e Registration of MBI 203

. Unpublished study prepared by Marrone Bio Innovations, 275 p ages

.

481129

- 02 Koivunen M

. 2010. Supplemental Product Identity Study, Secondary Metabolites (MBI

- 203). U npublished study prepared by Marrone Bio Innovations, 13 6 p age s

.

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

- D ay 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 matricari ae ), 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 Iner t 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, 2 7 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, 1 23 pages.

4 81307

- 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

Characte ristics Study. Unpublished study prepared by Marrone Bio Innovations,

21 p ages

.Chromobacterium subtsugae strain PRAA4

- 1 T Page 23 of 34 Biopesticides Registration Action Document

4 84407

- 01 Pitts K. 2011. MBI 203 EP: 7

- Month Storage Stability and Corrosion Characteristics Study . Unpublished study prepared by Marrone Bio Innovations,

21 p ages

.

B. Envir onmental Protection Agency Risk Assessment Memoranda

U.S. EPA. 20 11 a

. Chromobacterium subtsugae strain PRAA4

- 1 T

. 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 Assessm ent of Chromobacterium subtsugae strain

PRAA4

- 1 T (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 O rnamental Plants, Turf and Edible Crops, Memorandum from A. Gross and Z. Vaituzis, Ph.D. to J. Kausch dated July 7, 2011.

C. Other References

Martin PAW, Gundersen

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

sp. n ov., a betaproteoba cterium 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 (Heter optera: Pentatomidae) and corn rootworm (Coleoptera: Chrysomelidae). Journal of Economic Entomology 100(3):680

– 684.Chromobacterium subtsugae strain PRAA4

- 1 T Page 24 of 34 Biopesticides Registration Action Document

A PPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS (40 CFR PART 158

– SUBPART V )

TABLE 1. Product Analysis Data R equirements for the Technical Grade of the Active Ingredient (TGAI) /Manufacturing

- Use Product ( MP ) , MBI

- 203 TGAI (40 CFR § 158.2 120 )

OCSPP
G uideline

N umber Data Requirement Results MRID No. TGAI MP

885.1100 Product Identity Not applicable Submitt ed data fulfill the requirement for product identity. MBI

- 203 TGAI

contains 100

. 00 % by weight Chromobacterium subtsugae

strain PRAA4

- 1 T cells and spent fermentation media ( minimum of 1,5 00 Cabbage Looper Potency Units per milligram). 479450

- 01

481129

- 02

481 307

- 01

885.1200 Manufacturing Process Submitted data fulfill the requiremen t for manufacturing

process

. 479450

- 01

481129

- 03

Not a pplicable Deposition of a Sample in a Nationally Recognized Culture

Collection Chromobacterium subtsugae

strain PRAA4

- 1 T i s 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 B

30655. Not applicable 479450

- 01

885.1300 Discussion of Formation

of Unintentional Ingredients Submitted data fulfill the requiremen t for discussion of formation of unintentional ingredients

. 479450

- 01

885.1400 Analysis of S amples Submitted data fulfill the requirement for analysis of samples. 479450

- 01

8 85 .1 50 0 Cert ification of L imits Not applicable Limits listed o n the

confidential statement of

formula are adequate/a cceptable. 479450

- 01

Additional Studies

830.1800 Enforcement Analytical

M ethod Not applicable Submitted data fulfill the requiremen t for an

enforcemen t analytical

method

. 479450

- 01Chromobacterium subtsugae strain PRAA4

- 1 T Page 25 of 34 Biopesticides Registration Action Document

TABLE 2. Product Analysis Data Requirements for the End

- Use Product ( E P), MBI

- 203 EP (40 CFR § 158.2120)

OCSPP Guideline Number Data Requirement Results MRID No.

885.1100 Product Identity Submitt ed data fulfil l the requirement for product identity. MBI

- 203 EP contains 94.50% by weight Chromobacterium subtsugae strain PRAA4

- 1 T cells and spent fermentation media (minimum of 4,000 Cabbage Looper Potency Units per milligram). 479450

- 02

481129

- 06

885.1200 Manufact uring 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 Formati on 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

se ction 3(c)(5) registration. As a term of

the MBI

- 203 EP registration, EPA will require a five

- batch analysis , addressing all quality control parameters (i.e., dry weight, standard plate count, viscosity, density, and pH) , be submitted within

one year. 4794 50

- 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 analytic al method. 479450

- 02Chromobacterium subtsugae strain PRAA4

- 1 T Page 26 of 34 Biopesticides Registration Action Document

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
G uideline

N umber 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

E levated Temperatures, Metals,

and Metal I ons Expe cted 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 e mulsifiable 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

- 01Chromobacterium subtsugae strain PRAA4

- 1 T Page 2 7 of 34 Biopesticides Registration Action Document

TABLE 4. Physical and Chemical Characteristics for the End

- Use Produc t (EP), MBI

- 203 EP (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

Eleva ted 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 registration, EPA will require 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. A s a term of the MBI

- 203 EP registration, EPA will require 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 47 9450

- 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 thi s information with its application for MBI

- 203 EP, it is summarized appropriately in this table

.Chromobacterium subtsugae strain PRAA4

- 1 T Page 28 of 34 Biopesticides Registration Action Document

T ABLE 5

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

- Use Product (MP)
, MBI

- 203 TGAI (40 CFR § 158.2 140)

OCSPP Guideline

N umber Data Requirement Results MRID N o.

TGAI M P

Tier I

885.3050 Acute Oral Toxicity/Pathogenicity Waived based on Chromobacterium subtsugae

strain PRAA4

- 1 T ’s natural occurrence in the soil, a

literature search that did not

iden tify any reports of human or animal infection by Chromobacterium subtsugae

, and 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

Chrom obacterium subtsugae

strain PRAA4

- 1 T ’s natural occurrence in the soil, a

literature search that did not identify any reports of human or animal infection by Chromobacterium subtsugae

, and the results of MRID Nos.

479450

- 11 and 479450

- 05.

Classification: Acceptable Not applicable 479450

- 23

885.3200 Acute Injection Toxicity/Pathogenicity (Intravenous) N ot toxic, infective, and/or pathogenic to rats when dosed intravenously at 3.1 x 10 6 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 durin g research, development, or testing of the

TGAI /MP , were reported by the applicant. An y future hypersensitivity incidents must be reported per OCSPP Guideline

885.3400. 479450

- 23

885.3500 Cell Culture Not required because Chromobacterium subtsugae

strain PRAA4

- 1 T is not a virus (refer to test note #4 of 40 CFR § 158.2140(d)). Not applicab le Not applicable

870.1100 Acute Oral Toxicity Not applicable Oral LD 50 female rats > 5,000 mg/kg Classification: Acceptable
TOXICITY CATEGORY I V 479450

- 03

870.1200 Acute Dermal Toxicity Not applicable Dermal LD 50 c ombined (male and female) rats > 5,050 mg/kg

Classification: Acceptable
TOXICITY CATEGORY I V 479450

- 04

870.1300 Acute Inhalation Toxicity Not applicable Inhalation LC 50 combined (male and female) rats > 2.12 mg/L Classification: Acceptable
TOXICITY CATEGORY I V 479450

- 05Chromobacterium subtsugae strain PRAA4

- 1 T Page 29 of 34 Biopesticides Registration Action Document

T ABLE 5

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

- Use Product (MP)
, MBI

- 203 TGAI (40 CFR § 158.2 140)

OCSPP Guideline

N umber Data Requirement Results MRID N o.

TGAI M P

870.2400 Acute Eye Ir ritation Not applicable MBI

- 203 TGAI was minimally irritating to the eyes of rabbits. Classification: Acceptable
TOXICITY CATEGORY I V 479450

- 06

870.2500 Primary Dermal

Irritation Not applicable MBI

- 203 TGAI was slightly irritating to the skin of rabbits. Classification: Acceptable
TOXICITY CATEGORY I V 479450

- 07

Tiers II and III Not required for Chromobacterium subtsugae strain PRAA4

- 1 T based on the lack of acute toxicity/pathogenicity in the Tier I

studies.

Additional Studies

870.2600 Skin Sensitiza tion Not a dermal sensitizer Classification: Acceptable 479450

- 08

Not applicable Safety Study ( Subcutaneous

Injection ) Not toxic or pathogenic at > 10 6 cfu/mouse subcutaneously Classification: Supplemental

– Not a required study 479450

- 09

885.3200 Acut e Injection Toxicity/Pathogenicity

(Intravenous) Pilot Study Provides no useful data as to the mechanism of toxicity observed Classification: Supplemental

– Not a required study 479450

- 10Chromobacterium subtsugae strain PRAA4

- 1 T Page 30 of 34 Biopesticides Registration Action Document

TABLE 6 . Toxicology Data Requirements for the End

- Use Product (EP), MBI

- 203 EP (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 d uring research, development, or testing of the EP, were reported by the applicant. Any future hypersensitivity incidents must be reported per OCSPP Guideline

885.3400. 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 a re 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

- 0 6 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

- 08Chromobacterium subtsugae strain PRAA4

- 1 T Page 31 of 34 Biopesticides Registration Action Document

TABLE 6 . Toxicology Data Requirements for the End

- Use Product (EP), MBI

- 203 EP (40 CFR § 158.2140)

870.2500 Prima ry 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

- 1 T (40 CFR § 158.2150)

O CSPP 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 10 11 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 Fi sh 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

10 7 cfu/mL. Classification: Acceptable 479450

- 14

885.4240 Freshwater Invertebrate

Toxici ty/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 MBI

203 EP 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 MBI

- 203 EP label will (1) include a restriction that applications not occur within 150 feet of aquatic habitats and (2) list certain requirements and instructions to

further reduce potential for spray drift to offsite

areas

.Chromobacterium subtsugae strain PRAA4

- 1 T Page 32 of 34 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

- 1 T (40 CFR § 158.2150)

885.4280 Estuarine/Marine Fish and Invertebrate Testing Not req uired 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 CF R § 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 label will still require all users to test for phytotoxicity prior to pesticide application.

885.4340 Nontarget Insect Testin g 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.

Classi fication: Supplemental but acceptable

for risk assessmen t 479450

- 16

479450

- 17

479450

- 18

481129

- 05

The following mitigating language is required in

the Environmental Hazards section of the MBI

203 EP label: “This product is toxic to certain
nontarget ter restrial 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 label. Additionally, the MBI

- 203 EP la bel will list

c ertain 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 MBI

203 EP 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

label. Additionally, the MBI

- 203 EP label will list certain requirement s and instructions to further reduce potential for spray drift to offsite areas. Tiers II, III, and IV Not required for Chromobacterium subtsugae strain PRAA4

- 1 T based on the current uses and application methods.Chromobacterium subtsugae strain PRAA4

- 1 T Page 33 of 34 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

- 1 T (40 CFR § 158.2150)

Additional Studies Not applicable E ndangered 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 endangere d 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

– n ot a required study 479450

- 22Chromobacterium subtsugae strain PRAA4

- 1 T Page 34 of 34 Biopesticides Registration Action Document

APPENDIX B. PESTICIDE PRODUCTS

EPA File Symbol Registration

Name Percentage
Active Ingredient Formulation Type Use Site(s) Method(s) of Application Application

Rate Target

Pest

84059

- O MBI

- 203

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

84059

- RN MBI

- 203 EP

or Triple Crown 94.50% End Use

– Liquid Various agricultural

and greenhouse crops (e.g., vegetables, ornamental,

and turf) Standard ground or aerial spray application equipment

and

chemi gation 2

– 12 quarts

per acre Various

insects and

mites