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

September 27 , 2011Chromobacterium subtsugae strain PRAA4

- 1 T Page 2 of 36 Biopesticides Registration Action Document

TABLE OF CONTENTS

I. EXECUTIVE SUMMARY

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

II. ACTIVE INGRED IENT OVERVIEW

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

III. REGULATORY BACKGROUN D

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

A. Applications for Pesticide Product Registration

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

B. Food Tolerance Exemption

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

IV. RISK ASSESSMENT

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

A. Pro duct 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 JUSTIC E

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

VI. RISK MANAGEMENT DECI SION

................................ ................................ ............. 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 ABBR EVIATIONS

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

IX. BIBLIOGRAPHY

................................ ................................ ................................ ........... 21

A. Studies Submitted to Support the Chromobacterium subtsugae strain PRAA4

- 1 T

Pesticide Product Registrations

................................ ................................ ................................ ..... 21

B. Environmental Protection Agency Risk Assessment Memoranda and Other Documents

....... 24

C. Other References

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

APPENDIX A. MICROBI AL PESTICIDES DATA R EQUIREMENTS

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

APPENDIX B. PESTICI DE PRODUCTS

................................ ................................ ............... 36Chromobacterium subtsugae strain PRAA4

- 1 T Page 3 of 36 Biopesticides Registration Action Document

BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

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

Microbial Pesticides Branch

Science Revie ws

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 LeaderChromobacterium subtsugae strain PRAA4

- 1 T Page 4 of 36 Biopesticides Registration Action Document

I. EXECUTIVE SUMMARY

Chr omobacterium subtsugae strain PRAA4

- 1 T is a gram

- negative, violet

- pigmented bacterium

that was isola ted from soil under a n eastern hemlock ( Tsuga c anadensis ) 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 ( Leptino tarsa
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

- 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 mic robe’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

- 1 T , Marrone Bio Innovations proposed to register both a manu facturing

- use pesticide product, MBI

- 203 TGAI , and an end

- use pesticide product, MBI

- 203 EP Bioinsecticide (formerly MBI

- 203 EP) , containing thi s bacterium. MBI

- 203 EP Bioinsecticide will be used to control label

- specified insect and mite

p ests (e.g., Euro pean 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

- 1 T pesticide product s. 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)

.

P roduct analysis data requirements for Chromobacterium subtsugae strain PRAA4

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

characteri stics, were fulfilled by acceptable guideline studies.

Ade quate mamm alian toxicology data an d 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)

. Moreove r, no hypersensitivity incidents, occurring during research, development, or testing 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 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

- 1 T . No dietary risks are expected from use of Chromobacterium subtsugae strain PRAA4

- 1 T as a n active ingredient in pest icide products.Chromobacterium subtsugae strain PRAA4

- 1 T Page 5 of 36 Biopesticides Registration Action Document

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

- 1 T 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 con trol
agent.

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 ingredient, due to their occupation, for prolonged peri ods 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 In stitute 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 and data waiver rationale, submitted by the app licant 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 performed an environmental risk assessment based on the data and data waiver rationale provided by the applicant a nd determined that the use s of

Chromob acterium subtsugae strain PRAA4

- 1 T 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 sit es 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 soil

directed applications to crop pl ants 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 fo r 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 pe sticide products containing Chromobacterium subtsugae strain PRAA4

- 1 T

, 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 r eceived on EPA’s preliminary decision to register the Chromobacterium subtsugae strain PRAA4

- 1 T pesticide products , MBI

- 203 TGAIChromobacterium subtsugae strain PRAA4

- 1 T Page 6 of 36 Biopesticides Registration Action Document

and MBI

- 203 EP Bioinsecticide . Therefore, EPA maintained that, based upon the risk assessment and information submitted in supp ort 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

Chromob acterium 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 Pesticide: 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 products containing this active ingredient.

III. REGULATORY BACKG ROUND

A. Applications for Pesticide Product 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 pesticide product, MBI

- 203 TGAI (EPA File Symbol 84059

- O )

, and an end

- use pesticide 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 product 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 comments were received following this publicat ion.

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 tole rance for Chromobacterium subtsugae strain PRAA4

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

. In the Federal Register of March 10, 2010 ( 75 FR 11171 ), EPA

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

, Chromobacterium subtsugae strainChromobacterium subtsugae strain PRAA4

- 1 T Page 7 of 36 Biopesticides Registration Action Document

PRAA4

- 1 T

, 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

- 1 T in or on all food commodities when applied as an insecticide or miticide and used in acco rdance 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 dat a 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. T he 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 co ntained 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 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 must be submitted.

For the 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 pesti cide 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 )

A ll product analysis data requirements for Chromobacterium subtsugae strain PRAA4

- 1 T have

been fulfilled . Refe r 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

- 1 T have been

fulfilled

. Acceptable T ier I mammalian toxicology 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.Chromobacterium subtsugae strain PRAA4

- 1 T Page 8 of 36 Biopesticides Registration Action Document

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

in dividual 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; Mast er 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

- 1 T

. In a n 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, adverse ef fects, 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 material ( 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 necropsy an d 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, lu ngs, 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

- 1 T would not be toxic, infective, and/or pat hogenic through the oral route of exposure and that further testing is not necessary.

Acute Pulmonary Toxicity/Pathogenicity (OCSPP Gui deline 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

- 1 T

. 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, and 21 after treatment. Tissue and blood samples were collected at necropsy and cultured to provide qualitative and qu antitative 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 con tents by day 3. The test material was nontoxic and

noninfective. EPA believes these data, when taken together, clearly indicate that Chromobacterium subtsugae strain PRAA4

- 1 T would not be toxic, infective, and/or pathogenic through the inhalation route of exposure and that further testing is not necessary.Chromobacterium subtsugae strain PRAA4

- 1 T Page 9 of 36 Biopesticides Registration Action Document

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 Sprag ue

- 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 a s 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 b lood, 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 [M BI 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 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

- 1 T

, were reported by the applicant . Any future hypersensitivity i ncidents 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

- 1 T is not a virus (refer to test note #4 of 40 CFR § 158.2 140(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 fo r Chromobacterium subtsugae 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 Endo crine 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 end ocrine 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 wh ere EPA

will det ermine 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 pot ential for human exposure through pathways such asChromobacterium subtsugae strain PRAA4

- 1 T Page 10 of 36 Biopesticides Registration Action Document

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 st rain PRAA4

- 1 T 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 pesti cide chemicals. Accordingly, EPA anticipates issuing future EDSP

orders/data cal l

- 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 chemic al 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 resid ue, including all anticipated dietary exposures and all other exposures for which there is reliable information.” This includes exposure through drinking water and in

resi dential 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....” Additiona lly, 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 ana lyses to determine the risks from aggregate 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 in formation 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 informat ion discussed previously and presented in Table 5 in Appendix A , t he data required for a FFDCA risk assessment for Chromobacterium subtsugae strain PRAA4

- 1 T have been fulfilled

.Chromobacterium subtsugae strain PRAA4

- 1 T Page 11 of 36 Biopesticides Registration Action Document

a. Aggregate Exposure

In examining aggre gate 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 thr ough 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 st rain PRAA4

- 1 T 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 . Rat her, 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 m icrobial 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

- 1 T (see

secti on 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

- 1 T in consumed drinking water is unli kely. The proposed u se patterns for Chromobacterium subtsugae strain PRAA4

- 1 T 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 subt sugae strain

PRAA4

- 1 T 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 so il. I f Chromobacterium subtsugae strain PRAA4

- 1 T were to be transferred to surface or ground water s that are intended for eventual human consumption (e.g., through spray drift or runoff) and directed to wastewater treatment systems or drinking water facili ties

, it likely w ould not survive the cond itions water is subjected to in such systems or facilities , including high temperatures, chlorination, pH adjustments, and/or filtration (Center s for Disease Control and Prevention 2009; U.S. EPA 2004)

. In the remo te 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

- 1 T (see section IV(B)(1)(a) and Table 5 in Appendix

A )

.

Non

- occupational, Residential Risk Characterization : Dermal and inhalation non

- occ upational
exposure to Chromobacterium subtsugae strain PRAA4

- 1 T 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

- 1 T
is not toxic (acute inhalation and dermal toxicity), is only slightly irritating (primary dermal irritation), is not a sensitizer (dermal s ensitization), and is not pathogenic or infective (acute injection toxicity/pathogenicity) (see section IV(B)(1)(a) and Table 5 in Appendix A )

.Chromobacterium subtsugae strain PRAA4

- 1 T Page 12 of 36 Biopesticides Registration Action Document

b. Cumulative Effects from Substances with a Common Mechanism of Toxicity

Se ction 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 subst ances 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 substances, and Chromobacterium subtsugae strain

PRAA4

- 1 T 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

- 1 T does not have a common mechanism of toxicity with other substances. Following from this, therefor e, EPA concludes that there are no cumulative effects associated

with Chromobacterium subtsugae strain PRAA4

- 1 T that need to be considered. For information

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

and to eval uate the cumulative ef fects o f 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 sectio n 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 cumulat ive effects on infants and children of the residues and other substances with a common mechanism 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 eness 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 t he U.S. population, including infants and children, from aggregate exposure to the residues of Chromobacterium subtsugae strain PRAA4

- 1 T

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

informati on. 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

- 1 T do not demonstrate toxic, pathogenic

, and/ o r 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

.Chromobacterium subtsugae strain PRAA4

- 1 T Page 13 of 36 Biopesticides Registration Action Document

3. Oc cupational Exposure and Risk Characterization

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 pesticid e product label s to mitigate any potential risks to pesticide handlers due to prolonged or numerous exposure s . Handlers applying Chromobacterium subtsugae strain

PRAA4

- 1 T end

- use pesticide products in agricultural settings must wear a long

- sleeved shirt,

l ong 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 Healt h 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 effec ts to the U.S. population in general, and to infants and 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 pro ducts containing Chromobacterium subtsugae strain PRAA4

- 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 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

- 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 terrestrial arthropods, aquatic invertebrates, and honey bees. Use sites and application methods, i n 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 Exposur e 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 t o 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 (OC SPP 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 andChromobacterium subtsugae strain PRAA4

- 1 T Page 14 of 36 Biopesticides Registration Action Document

spe nt fermentation media) to northern bobwhite ( 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 fiv e 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 Chromobacteri um subtsugae strain PRAA4

- 1 T

.

In an 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 om 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 toxicity/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 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

fulfille d testing requirements was submitted ; ther e were no observed effects from test

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 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

- 1 T 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

- 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 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 MB I

- 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 ar e visiting the treatment area.Chromobacterium subtsugae strain PRAA4

- 1 T Page 15 of 36 Biopesticides Registration Action Document

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 bot h 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 is not related to any known plant pathogen, effe cts 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 Inverte brates (OCSPP Guideline 885.4280) : MBI

203 EP Bioinsecticide (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 demonstrated toxici ty 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 testin g 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

. 47945 0

- 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 ratio nale 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 con centrations.

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. D rift and runoff may be hazardous to aquatic

orga nisms 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, marshe s, ponds, estuaries, and commercial fish ponds).Chromobacterium subtsugae strain PRAA4

- 1 T Page 16 of 36 Biopesticides Registration Action Document

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 Literatu re 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 Environme ntal 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 Technol ogy; 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 we re two documents published by the United States Department of Agriculture

– Agricultural Research Service concerning the toxicity of Chromobacterium subtsugae to
Colorado p otato b eetle and other insect pests (Martin et al . 2007a) , as well as to s outhern g r een

s tink b ug and c orn r ootwor m (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 t he 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

asse ssment for Chromobacterium subtsugae strain PRAA4

- 1 T , f urther testing at higher tier levels (i.e., T iers 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 app lications (aerial sprays and chemigation) of Chromobacterium subtsugae strain 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 spec ific 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
tha n 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 inverte bra tes 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 PR AA4

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

- 203 EP Bioinsecticide label

.Chromobacterium subtsugae strain PRAA4

- 1 T Page 17 of 36 Biopesticides Registration Action Document

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

- 1 T to the proposed crops:

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

BPPD concludes that the following types of labeled application s of Chromobacterium subtsugae

strain PRAA4

- 1 T to the proposed crops have the potential to cause direct effects, indirect effec ts, 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 chemigat ion 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, imple mentation, 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 re sulting 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 oppo rtunity 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 i n 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 Chromobac terium subtsugae strain PRAA4

- 1 T , 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

.Chromobacterium subtsugae strain PRAA4

- 1 T Page 18 of 36 Biopesticides Registration Action Document

V I

. RISK MANAGEMENT DECISION

Section 3(c)(5) of FIFRA permits for the registration of a pesticide provided that all the following d eterminations 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 u nreasonable 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

- 1 T 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 label s, and such products are not expected to

cause unrea sonable adverse effects on the environment when used according to the label instructions. Criterion 2 is satisfied by the current product label s, as well as the data and

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

PRAA4

- 1 T 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, s atisfying 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 E P Bioinsecticide

, 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., a quatic 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 Labeling

Before releasing pesticide products containing Chromobac terium 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 Bioinsecticide registration, the registrant must submit the following

dat a 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).Chromobacterium subtsugae strain PRAA4

- 1 T Page 19 of 36 Biopesticides Registration Action Document

(2) Storage Stability (OC SPP 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 type s 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 conf irmed incidents) must be reported to EPA under the provisions of 40 CFR § 158.2140(d).Chromobacterium subtsugae strain PRAA4

- 1 T Page 20 of 36 Biopesticides Registration Action Document

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

ASAE American Society of Agricultural Engineers

BPPD Biopesticides and Pollution Prevention Division

BRAD Biopesti cides 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 Disruptor Screening Program

EP end

- use product

EP A 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

LC 50 median lethal concentr ation. 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 m edian 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 technical grade of the active ingredient

U.S. United StatesChromobacterium subtsugae strain PRAA4

- 1 T Page 21 of 36 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 MBI

- 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

Procedu re 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

– Final Report. Project Number: 12780/09 . Unpub lished 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: A cute 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 Sensitization Study in Guinea Pigs

– Final Report. P roject 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 . Unpublished study prepared by Stillmeadow,

Incor porated

, 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 22 of 36 Biopesticides Registration Action Document

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

p ages

.

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 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 . Unpubl ished 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 Marrone Bio Innovations, 9 p ages

.

479450

- 1 7 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 Tox icity 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. 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 p ages

.

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 me llifera 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 Technolo gy Sciences Group, Incorporated, 23 p ages

.Chromobacterium subtsugae strain PRAA4

- 1 T Page 23 of 36 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, Incorporated, 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 the Registration of MBI 203

. Unpublished st udy 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 ages

.

481129

- 03 Chanbusarakum L

. 2010. Supp lemental 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, 2 7 pages.

48 1129

- 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.

481307

- 01 Chanbusarakum L

. 2010. MBI 203 T GAI: 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 p ages

.Chromobacterium subtsugae strain PRAA4

- 1 T Page 24 of 36 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. Environmental Protection Agency Risk Assessmen t Memoranda and Other Documents

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 Assessment of Chromobacteriu m 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 Ornamental Plants, Tur f 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. 20 11d. 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 Ne w Zealand soils. Australian Journal of Soil Research

39:1397

- 1406.

Centers for Disease Control and Prevention. 2009. Drinking Water

- Water Treatment. Available

from h ttp://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. n ov., a betaproteobacterium toxic to Colorado po tato 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 co rn rootworm (Coleoptera: Chrysomelidae). Journal of Economic Entomology 100(3):680

– 684.

Pang L, M cLeod M, Aislabie J, Šimůnek J, Close M, Hector R. 2008. Modeling transport of
microbes in ten undisturbed soils under effluent irrigation. Vadose Zone Journa l 7:97

- 111.Chromobacterium subtsugae strain PRAA4

- 1 T 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 Tes t Guidelines

– Background for Residue Analysis of Microbial Pest Control Agents (OPPTS 885.2000). Available from http://www.epa.gov/ocspp/pubs/frs/publicatio ns/Test_Guidelines/series885.htm

.Chromobacterium subtsugae strain PRAA4

- 1 T Page 26 of 36 Biopesticides Registration Action Document

A PPENDIX 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 § 158.2 120 )

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 TG AI

contains 100

. 00 % by weight Chromobacterium subtsugae

strain PRAA4

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

- 01

481129

- 02

481307

- 01

885.1200 Manufacturing Process Submitte d 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 is on deposit with the Agricultural

Research Servi ce 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 fulf ill 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 Certification 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 enforcement analytical

method

. 479450

- 01Chromobacterium subtsugae strain PRAA4

- 1 T Page 27 of 36 Biopesticides Registration Action Document

TABLE 2. Product Analysis Data Requirements for the End

- Use Product ( E P), MBI

- 203 EP Bioinsecticide (40 CFR § 158.2120)

OCSPP Guideline Number Data Requirement Results MRID No.

885.1100 Product Identity Submitt ed data fulfill the requirement for product ide ntity. MBI

- 203 EP

Bioinsecticide contains 94.50% by weight Chromobacterium subtsugae strain

PRAA4

- 1 T 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 five

batch 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 enforcemen t analytical method. 479450

- 02Chromobacterium subtsugae strain PRAA4

- 1 T Page 28 of 36 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 MR ID 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 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 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 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 Stab ility 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 t o 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

83 0.7000* 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

- 1 T Page 30 of 36 Biopesticides Registration Action Document

T ABLE 5

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

- Use Product (MP)
, MBI

- 203 TGAI (40 CFR § 158.2140)

OCSPP Guideline

N umber Data Requirement Results MRID N o.

TGAI M P

Tier I

885.3050 Acute Oral Toxicity/Pathogenicity Waived based on the results of

MRID Nos. 479450

- 11 and

4794 50

- 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 Inject ion 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 in cidents, 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. 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

- 1 T 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 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

- 05

870.2400 Acute Eye Irritation Not applicable MBI

- 203 TG AI 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

TOXICIT Y 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.Chromobacterium subtsugae strain PRAA4

- 1 T Page 31 of 36 Biopesticides Registration Action Document

T ABLE 5

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

- Use Product (MP)
, MBI

- 203 TGAI (40 CFR § 158.2140)

OCSPP Guideline

N umber Data Requirement Results MRID N o.

TGAI M P

Additional Studies

870.2600 Skin Sensitization Not a dermal sensitizer

Cl assification: 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 Acute Injection Toxicity/Pathogenici ty

(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 32 of 36 Biopesticides Registration Action Document

TABLE 6 . Toxicology Data Requirements for the End

- Use Product (E P), 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 appli cable

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.

Classi fication: 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 iner t 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

48112 9

- 08Chromobacterium subtsugae strain PRAA4

- 1 T Page 33 of 36 Biopesticides Registration Action Document

TABLE 6 . Toxicology Data Requirements for the End

- Use Product (E P), 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

- 1 T (4 0 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 10 11 cfu/kg body weight/day for 5 days. Classification: Acceptable 47945 0

- 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 r equired 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

88 5.4200 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

10 7 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 MBI

203 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 MBI

203 EP Bioinsecticide label will (1) bear a maximum application rate of 12 quarts of product per acre, (2 ) include a restriction that aerial

a pplications not occur within 75 feet of aquatic

habitats , (3) require aer ial applicators to use

medium or coa rser spray nozzles, and ( 4 ) list certain requirements and instructions to further

reduce potential for spray drift to offsite areas

.Chromobacterium subtsugae strain PRAA4

- 1 T 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

- 1 T (4 0 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 wi ll 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 ef fects 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 beetl es. 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 r isk 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 Bioinsecticide label: “This product is toxic to certain nontarget terrestrial arthropods.

Minimiz e 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 la bel 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

- 2 0

479450

- 21

The following mitigating language is required in

the Environmental Hazards section of the MBI

203 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 Bio insecticide label will list certain requirements and instructions to further reduce potential for spray drift to offsite areas.Chromobacterium subtsugae strain PRAA4

- 1 T 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

- 1 T (4 0 CFR § 158.2150)

OCSPP Guideline

Number Data Requirement Results MRID No.

Tiers II, III, and IV Not required for Chromobacterium subtsugae strain PRAA4

- 1 T based on the current uses and application me thods.

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

co ncern, 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 det ermination. Classification: Supplemental

– not a required study 479450

- 22Chromobacterium subtsugae strain PRAA4

- 1 T 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 Ap plication 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,

ornamental s
,

and turf) Standard ground or aerial spray application equipment

and chemigation 2

– 12 quarts

per acre Various

insects and

mites