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

DRAFT

- March 1 6 , 2012Chromobacterium subtsugae strain PRAA4

- 1 T Page 2 of 41 Biopesticides Registration Act ion Document

TABLE OF CONTENTS

I. EXECUTIVE SUMMARY

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

II. ACTIVE IN GREDIENT OVERVIEW

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

III. REGULATORY BACKGROUN D

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

A. Applications for Pesticide Product Registration

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

B. Food Tolerance Exemption

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

IV. RISK ASSESSMENT

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

A. Product Analysis Assessment (40 CFR § 158.2120)

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

B. Human Health Assessment (40 CFR § 158.2140)

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

C. Environme ntal Assessment (40 CFR § 158.2150)

................................ ................................ ........ 14

V. ENVIRONMENTAL JUSTIC E

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

VI. RISK MANAGEMENT DECI SION

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

VII. ACTIONS REQUIRED O F THE REGISTRANT

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

A. Final Printed Labeling

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

B. Terms of Registration

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

C. Reporting of Adverse Effects and Hypersensitivity Incidents

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

VIII. GLOSS ARY OF ACRONYMS AND ABBREVIATIONS

................................ ......... 22

IX. BIBLIOGRAPHY

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

A. Studies Submitted to Support the Chromobacterium subtsugae st rain PRAA4

- 1 T

Pesticide Product Registrations

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

B. Environmental Protection Agency Risk Assessment Memoranda and Other Documents

....... 27

C. Other References

................................ ................................ ................................ ............................. 27

APPENDIX A. MICROBI AL PESTICIDES DATA R EQUIREMENTS

............................ 29

APPENDIX B. PESTICI DE PRODUCTS

................................ ................................ ............... 41Chromobacterium subtsugae strain PRAA4

- 1 T Page 3 of 41 Biopesticides Registration Act ion Document

BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

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

Science R eviews

Judy Facey, Ph.D. Product Analysis, Human Health Joel V. Gagliardi , Ph.D. Product Analysis, Human Health
John L. Kough, Ph.D. Product Analysis, Human Health Shannon Borges Environmental Effects
Anna Gross Environm ental Effects Zigfridas Vaituzis, Ph.D. 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 41 Biopesticides Registration Act ion Document

I. EXECUTIVE SUMMARY

Background Chromobacterium subtsugae strai n 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 Catoctin Mountain region of central Maryland. The United States Department of Agriculture found this
isolate of Chromob acterium subtsugae to be orally toxic to Colorado potato beetle ( Leptinotarsa
decemlineata ) larvae, small hive beetle ( Aethina tumida ) larvae, southern corn rootworm ( Diabrotica undecimpunctata ) larvae and adults, and southern green stink bug ( Nezara virid ula )
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 mot h ( Plutella xylostella ), and southern corn rootworm , suggesting this microbe’s insecticidal activity is due to reduction in weight or inhibition of feeding. In light of the demonstrated insecticidal and
miticidal capabilities of Chromobacterium subtsugae s train PRAA4

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

- use pesticide product, MBI

- 203 TGAI , and three end

- use pesticide product s , MBI

- 203 EP Bioinsecticide (formerly MBI

- 203 EP) , MBI

- 203 SC, and
GrandevoTM , containing thi s bacterium. M BI

- 203 EP Bioinsecticide , MBI

- 203 SC, and
GrandevoTM will be used 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, bedd ing plants, ornamentals, and turf , as well as on certain plants in residential settings

.

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 four
aforementioned Chromobacterium subtsugae strain PRAA4

- 1 T pesticide products. Overall, such
data and information are adequate for risk assessment purpo ses , fulfill current data requirements

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

.

Product Analysis

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.

Toxicology Adequate mamm alian toxicology data and information were submitted to support the
Chro mobacterium 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 duri ng 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/pathogenic ity tests, and the absence of hypersensitivity incidents, testing at higher tiers (i.e., Tiers II and III) was not required.Chromobacterium subtsugae strain PRAA4

- 1 T Page 5 of 41 Biopesticides Registration Act ion Document

Tolerance Exemption

EPA concluded that there is a reasonable certainty that no harm will result to the United States population, i ncluding 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 an active ingredient in pesticide products. 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 b e present on food or in drinking water, the acute oral toxicity and pathogenicity data/information demonstrated no toxicity, infectivity, and/or pathogenicity is likely to occur with any exposure level of this microbial pest control
agent.

Occupational Ex posure 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 periods or numerous times . Handlers working
with Chromobacterium subtsugae strain PRAA4

- 1 T in agricultural or commercial settings are directed to wear a long

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

Nontarget Organisms
The data and data waiver rat ionale, 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

nontar get organisms at higher tier levels (i.e., Tiers II, III, and IV) is not required for the current uses and application methods. EPA performed environmental risk assessment s , based on the data and data waiver rationale provided by the applicant

, a nd determi ned that the use s of Chromobacterium 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 invertebra tes, and honey bees. Use sites and application methods, in combination with mitigating label language (see Table 11 in Appendix A ), however, will limit exposure

. EPA has made “no effect” determinations for direct effects, ind irect 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 m ade 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

.

Public Participation On October 1, 2009, EPA announced a new policy to provide a more meaningful opportunity for the public to participate in major registration decisions before they occur. According to this policy, EPA intends to provide a public comment period prior to making a registration decisionChromobacterium subtsugae strain PRAA4

- 1 T Page 6 of 41 Biopesticides Registration Act ion Document

for, at minimu m, the following types of applications: new active ingredients; first food uses; first outdoor uses; first residential uses; or any other registration actions for which EPA believes there may be significant public interest.

Consistent with the policy of making registration actions more transparent, in 2011, two pe sticide products containing Chromobacterium subtsugae strain PRAA4

- 1 T

, a new active ingredient at the time

, and allowing for this active ingredient’s first outdoor and food uses were subject to a 30

- day comment period. During this comment period, no comments were received on EPA’s preliminary decision to register the Chromobacterium subtsugae strain PRAA4

- 1 T pesticide products , MBI

- 203 TGAI and MBI

- 203 EP Bioinsecticide . Therefore, EPA maintained that, based upon the risk assessment and information submitted in support of registration of such pesticide products, it was appropriate to issue the MBI

- 203 TGAI and MBI

- 203 EP
Bioinsecticide registrations

.

Marrone Bio Innovations has now request ed to register a pesticide product that is labeled for
Chromobacterium subtsugae strain PRAA4

- 1 T ’ s first residential uses. Thus, EPA is opening
another 30

- day comment period for public participation for this particular pesticide product

— GrandevoTM

. The docket ide ntification number, associated with this registration action and accessed through either http://www.regulations.gov or http://www.e pa.gov/pesticides/regulating/registration

- status.html is EPA

- HQ

- OPP

- 2010

- 0058. The following documents are available for comment in EPA

- HQ

- OPP

- 2010

- 0058: (1) draft Chromobacterium subtsugae strain PRAA4

- 1 T Biopesticides Registration Action Document (BRAD) (dated March 2012) and (2) draft label for the end

- use pesticide product, GrandevoTM (EPA File Symbol 84059

- RT ).

II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Chromobacterium subtsugae strain PRAA4

- 1 T

Culture Deposit: Agricultural Research Serv ice 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

– Insect icide and Miticide

See Appendix B for specific information (e.g., use sites, application rates, methods of application, formulation types, and target pests) regarding the registered and proposed pesticide products cont aining
this active ingredient.Chromobacterium subtsugae strain PRAA4

- 1 T Page 7 of 41 Biopesticides Registration Act ion Document

III. REGULATORY BACKGROUND

A. Applications for Pesticide Product Registration

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

- 107; Davis, California 95618) submitted application s t o 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 regi ster 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 publication.

On December 9, 2010, Marrone Bio Innovations submitted an application to register an end

- use pesticide product, MBI

- 203 SC (EPA File Symbol 84059

- RA) under FIFRA section 3. On February 2, 20 11, EPA announced receipt of this application to register a pesticide product containing a new active ingredient ( 76 FR 5805 ) and opened a 30

- day public comment period
pursuant t o the provisions of FIFRA section 3(c)(4). No comments were received following this publication.

On September 30, 2011, Marrone Bio Innovations submitted an application to register an end

- use pesticide product, Grandevo TM (EPA File Symbol 84059

- RT) under FIFRA section 3. On February 15, 2012, EPA announced receipt of this application requesting to register a new use (i.e., residential uses) ( 77 F R 8861 ) for Chromobacterium subtsugae strain PRAA4

- 1 T and opened a 30

- day public comment period pursuant to the provisions of FIFRA section 3(c)(4). No comments were received following this publication.

B. Food Tolerance Exemption

Concurrent with its M BI

- 203 TGAI and MBI

- 203 EP 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
subts ugae strain PRAA4

- 1 T (Pesticide Petition (PP) 9F7674 ). In the Federal Register of March 10, 2010 ( 75 FR 11171 ), EPA announced that Marrone Bio Innovations proposed to establish a n 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

- day comment period. No comments were received following this publication.

On Septem ber 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 accordance with good agricultural p ractices (40 CFR § 180.1305; 76 FR 55272 ).Chromobacterium subtsugae strain PRAA4

- 1 T Page 8 of 41 Biopesticides Registration Act ion Document

IV. RISK ASSESSMENT

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

eva luated for this BRAD were considered in light of these requirements.

The classifications that are found for each data submission are assigned by EPA science
reviewers and are an indication of the usefulness of the information contained in the documents
fo r 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 cert ain 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 i ndicates 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 information submitted to EPA in support of a pesticide registration. The acti ve 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. Refer to Tables 1, 2, 3, 4

, 5, 6, and 7 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 Tier I mammalian to xicology 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 to xicity/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 registra tions, refer to Tables 8, 9, and 10 in Appendix A

.Chromobacterium subtsugae strain PRAA4

- 1 T Page 9 of 41 Biopesticides Registration Act ion Document

a. Acute Toxicity/Pathogenicity

– Tier I

Acute Oral Toxicity/Pathogenicity ( Harmonized Guideline 885.3050; Master Record Identification Number (MRID No.) 479450

- 23) : Up on 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 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, adverse 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 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 and cultured to provide qualitative and quantitative measur ements of the test organism. There was no mortality during the study, and all rats appeared normal throughout. The test organism had cleared completely from blood, kidneys, mesenteric lymph nodes, lungs, brain, liver, spleen, and cecum contents by day 3. T he 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 oral route of exposure and that furth er testing is not

necessary.

Acute Pulmonary Toxicity/Pathogenicity ( Harmonized 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/p athogenicity 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 n ose

- 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 con centration (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 fr om the active test material group were sacrificed and necropsied on days 0, 3, 7, 14, and 21 after treatment. Tissue and blood samples were collected at necropsy and cultured to provide qualitative and quantitative measurements of the test organism. There was no mortality during the study, and all rats appeared normal throughout. The test organism had cleared completely from blood, kidneys, mesenteric lymph nodes, lungs, brain, liver, spleen, and cecum contents by day 3. The test
material was nontoxic and n oninfective. 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.

Acute Injection Toxicity/Pathogenicity (Intravenous)

– Rat ( Harmonized Guideline 885.3200;
MRID No. 479450

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

- Dawley rats were injected at 3.1 x 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. NoChromobacterium subtsugae strain PRAA4

- 1 T Page 10 of 41 Biopesticides Registration Act ion Document

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, mese nteric 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 ( Harmonized Guideline 885.3400; MRID No. 479450

- 23) : No hypersensitivity incidents, including immediate

- type or del ayed

- 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 to EPA (refer to test note #3 of 40 CFR § 158.2140(d))

.

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

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

b. Acute Toxicology and Su bchronic Toxicity/Pathogenicity

– Tier II; Reproductive Fertility Effects, Carcinogenicity, Immunotoxicity, and Infectivity/Pathogenicity Analysis

– Tier III

Tier II and Tier III studies were not required for Chromobacterium subtsugae strain PRA A4

- 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 (ED SP) to determine whether certain substances (including pesticide active and other ingredients) may have an effect in humans or wildlife similar to an effect produced by a “naturally occurring estrogen, or other such endocrine effects as the Administrator m ay 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 thyro id (E, A, or T) hormonal systems. Chemicals that go through Tier 1 screening and are found to have the potential to interact with E, A, or T hormonal systems will proceed to the next stage of the EDSP where EPA will determine which, if any, of the Tier 2 t ests 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 200 9 and February 2010, EPA issued test orders/data call

- ins for the first group of 67 chemicals, which contains 58 pesticide active ingredients and 9 inert ingredients. This list of chemicals was selected based on the potential for human exposure through pat hways 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 among the group of 58 pesticide active ingredients on the initial list to be screened under the EDSP. Under FFDCA section 408(p), EPAChromobacterium subtsugae strain PRAA4

- 1 T Page 11 of 41 Biopesticides Registration Act ion Document

must screen all pesticide chemicals. Accordingly, EPA anticipates issuing future EDSP
orders/data call

- ins for all pesticide active ingred ients.

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/e ndo/

.

2

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

Section 408(c)(2)(A)(i) of FFDCA allows EPA to establish an exemption from the requirement for a tolerance (the legal limit for a pesticide chemical residue in or on a food) only if E PA determines that the exemption is “safe.” Section 408(c)(2)(A)(ii) of FFDCA defines “safe” to mean that “there is a reasonable certainty that no harm will result from aggregate exposure to the pesticide chemical residue, including all anticipated dietary exposures and all other exposures for which there is reliable information.” This includes exposure through drinking water and in
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....” Additionally, section 408(b)(2)(D) of FFDCA re quires 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 agg regate exposure to pesticide residues. First, EPA determines the toxicity of pesticides. Second, EPA examines exposure to the pesticide through food, drinking water, and through other exposures that occur as a result of pesticide use in residential setting s.

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

Based on the acute toxicity/pathogenicity data and information discussed previously and
presente d in Table 8 in Appendix A , t he data required for a FFDCA risk assessment for Chromobacterium subtsugae strain PRAA4

- 1 T have been fulfilled

.

a. Aggregate Exposure

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

Food Exposure and Risk Characterization : Any exposure to this naturally occurring soilChromobacterium subtsugae strain PRAA4

- 1 T Page 12 of 41 Biopesticides Registration Act ion Document

bacterium is anticipated to be negligible. Although Chromobacterium subtsugae strain 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 . Rather, after application, it likely will
d egrade due to predation by other biological organisms (e.g., protists) and exposure to particular environmental factors (e.g., sunlight and varying temperatures) (Lindow and Brandl 2003 ; U.S.
EPA 1996 )

. Should this microbial pesticide be present on food, t he 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 8 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 unlikely. The proposed use patterns for Chromobacterium subtsuga e 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 subtsugae 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 soil. 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 facilities

, it likely w ould not survive the co nd 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 remote likelihood that this microbial pestic ide 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 le vel of Chromobacterium subtsugae strain PRAA4

- 1 T (see section IV(B)(1)(a) and Table 8 in Appendix

A )

.

Non

- occupational, Residential Risk Characterization : Dermal and inhalation non

- occupational
exposure to Chromobacterium su btsugae strain PRAA4

- 1 T may occur because some of the proposed pesticide applications would take place in residential settings . Even if dermal and
inhalation non

- occupational exposures were to occur, such exposures would not exceed EPA’s level of concern g iven 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 sensitization), and is not pathogenic or in fective (acute injection toxicity/pathogenicity) (see section IV(B)(1)(a) and Table 8 in Appendix A ).

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

Section 408(b)(2)(D)(v) of FFDCA requires tha t, when considering whether to establish, modify, or revoke a tolerance exemption, EPA consider “available information concerning the cumulative effects of [a particular pesticide’s]

. . . residues and other substances that have a common
mechanism of toxic ity.”

EPA has not found Chromobacterium subtsugae strain PRAA4

- 1 T to share a common mechanism of toxicity with any other substances, and Chromobacterium subtsugae strainChromobacterium subtsugae strain PRAA4

- 1 T Page 13 of 41 Biopesticides Registration Act ion Document

PRAA4

- 1 T does not appear to produce a toxic metabolite produced by other substances . F or 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, therefore, EPA concludes that there are no cumulati ve effects associated
with Chromobacterium subtsugae strain PRAA4

- 1 T that need to be considered. For information regarding EPA ’s efforts to determine which chemicals have a common mechanism of toxicity
and to evaluate the cumulative effects o f such chemica ls, see EPA ’s website at http://www.epa.gov/pesticides/cumulative

.

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

FFDCA section 408(b) (2)(C) provides that EPA shall assess the available information about consumption patterns among infants and children, special susceptibility of infants and children to pesticide chemical residues, and the cumulative effects on infants and children of the residues and other substances with a common mechanism of toxicity. In addition, FFDCA section 408(b)(2)(C) provides that EPA shall apply an additional tenfold (10X) margin of safety for infants and children in the case of threshold effects to account for p renatal and postnatal toxicity and the completeness of the database on toxicity and exposure unless EPA determines that a different margin of safety will be safe for infants and children. This additional margin of safety is commonly referred to as the Food Quality Protection Act Safety Factor. In applying this provision, EPA either retains the default value of 10X or uses a different additional safety factor when reliable data available to EPA support the choice of a different factor.

Based on the acute tox icity and pathogenicity data /information summarized in section IV(B)(1)(a) and Table 8 in Appendix A

, EPA concludes that there is a reasonable certainty that
no harm will result to the U.S. population, including infants and c hildren, 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
information. EPA has arrived at this conclusion beca use , 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 p opulations 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

.

3. Occupational 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 pesticide product labels to mitigate any potential ris ks to pesticide handlers due to prolonged or numerous exposures. Handlers applying Chromobacterium subtsugae strain

PRAA4

- 1 T end

- use pesticide products in agricultural or commercial settings must wear a long

sleeved shirt, long pants, socks, shoes, waterpr oof 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.Chromobacterium subtsugae strain PRAA4

- 1 T Page 14 of 41 Biopesticides Registration Act ion Document

4. Human Health Risk Characterization

EPA con sidered 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 unreasonable adverse effects to the U.S. population in ge neral, 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 products containing Chromobacteriu m 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 environmental risk assessment s 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 ris k to nontarget organisms when used according to label directions. Based on study results, there are concerns of toxicity to terrestrial arthropods, aquatic invertebrates, and honey bees. Use sites and application methods, in combination with mitigating lab el language, however, will limit exposure.

For a comprehensive summary of the generic data requirements described in sections IV(C)(1),
refer to Table 11 in Appendix A

.

1. Ecological Exposure and Risk Characterization

a. Terrestrial Animals and Plants

The end

- use product s (EP s ) are labeled for foliar spray , in

- furrow spray, and soil drench with ground or aerial applications. Aerial applications would increase exposure to nontarget insects and honey bees and, due to p otential 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 ( Harmonized Guideline 885.4050) and Mammals ( Harmon ized 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 bob white ( Colinus virginianus ). Fourteen

- day

old chicks were dosed with the test material (4.0 x 10 11 colony

- forming units per kilogram ( cfu/kg ) body weight), attenuated test material, or sterile saline once a day for five days and
observed for an additional 25 days. No signs of toxic or pathogenic effects were observed throughout the test or at necropsy. This study demonstrates that toxicity/pathogenicity to birds is not expected from labeled applications of Chromobacterium subtsugae strain PRAA4

- 1 T

.

In an a cute 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 room

t emperature prior to dosing. The animals were observed for 14 days. All animals survived,Chromobacterium subtsugae strain PRAA4

- 1 T Page 15 of 41 Biopesticides Registration Act ion Document

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 ( Harmonized Guideline 885.4340) and Honey Bees ( Harmonized Guideline 885.4380)

i) Data Submitted with Registration Appli cations for MBI

- 203 TGAI and MBI

- 203 EP

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 (MR ID 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 ; there 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 strai n 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 toxi city to honey bees with the use of Chromobacterium subtsugae
strain PRAA4

- 1 T (MRID No s

. 479450

- 19, 479450

- 20 and 479450

- 21).

ii) Data Submitted with Registration Application for MBI

- 203 SC

MRID No. 482844

- 02 : A laboratory study was conducted to assess the ac ute toxicity of MBI

- 203 TGAI when administered in the diet to adult ladybird beetles ( Hippodamia convergens ). A dose stated to be 10X the field application rate (4 quarts per acre) was sprayed onto corn earworm ( Helicoverpa zea ) eggs, which were provided a s diet. The test also included an inactivated MBI

203 TGAI group and an untreated control group. The beetles were observed for signs of adverse effects and egg consumption. It is not possible to confirm the dose rate; however, based on the dosage informati on provided, the concentration of the test solution was approximately 9.4 x 10 7 Cabbage Looper Potency Units per liter ( CLPU/L ) , which is comparable to the concentration of

MBI

- 203 SC at its lowest application rate. The application amount in the study was also stated to be “approximately” 0.3 milliliters ( mL ) in 5 mL deionized water, which introduces some additional uncertainty, especially at such a small volume. Nonetheless, the amount to which the ladybird beetles were exposed is very likely to be at leas t equal to the expected environmental concentration for nontarget insects at the lowest application rate. Since the dose rate cannot be confirmed, the study is classified as supplemental but is useful for ecological risk assessment.

MRID No. 482844

- 01 : I n this study, honey bees were exposed to a topical application of MBI

203 TGAI. An inactivated MBI

- 203 TGAI group, vehicle control groups (deionized water or acetone), and a positive control group (dimethoate) were also included in the study. Anesthetized bees were individually treated with 2 microliters ( μL ) of the appropriate solution and observedChromobacterium subtsugae strain PRAA4

- 1 T Page 16 of 41 Biopesticides Registration Act ion Document

for 48 hours. At test end, mortality in the test material groups was 5.0% when deionized water was the vehicle and 1.7% when acetone was the vehicle, which was similar to mortality observed in respective controls. The study is scientifically sound but is determined to be supplemental
because it is not of sufficient duration to determine pathogenicity and testing was not performed at a high dose. The 48

- hour conta ct LD 50 for MBI

- 203 in this study was greater than 25 micrograms (μg) active ingredient /bee. Based on the reported activity of the test material (at least
1500 Cabbage Looper Potency Units per mg ( CLPU/mg ) ), this is estimated to be 37.5 Cabbage Looper Pote ncy Units ( CLPU ) /bee. There is no standard determinant of toxicity for microbial pesticides for bee contact toxicity, so the meaning of this value in terms of risk is not known. The
concentration of the test substance used on the bees is estimated at 1.88 x 10 7 CLPU/L. Based on the proposed label dated August 21, 2011, MBI

- 203 SC is to be applied at up to 12 quarts in at least 10 gallons of water per acre. The resulting concentration of the spray that would be applied
based on these instructions is 8.1 x 10 7 CLPU/L to 2.4 x 10 8 CLPU/L (using the estimate of density from MRID 483335

- 01). These concentrations are above the concentration tested, so the results of this study may not be relevant to field exposures for the current proposed label.

MRID No. 48333 5

- 04 : This study was conducted to assess the acute topical/oral toxicity of MBI

203 TGAI (active ingredient, 100.0% Chromobacterium subtsugae strain PRAA4

- 1 T cells and spent fermentation media) when administered as food to cells containing honey bee larvae . The
test also included an inactivated MBI

- 203 TGAI group and a sucrose

- only control. Individual larval cells were treated with a 2 μL drop of the active or inactive test material in a 50% weight per volume sucrose solution, providing a dose of 100 μg act ive ingredient/bee. There were no adverse physiological responses noted in the test material or control group bees; however, this study has several deficiencies. Information about the conduct of the experiment was lacking. More importantly, different larva l stages of bees were used for different treatments, in that the larvae receiving the inactivated test material were younger than larvae in other treatments by several days. Worker bees aborted most of the larvae in this treatment, and the results may indi cate a methodological problem with the study or potential toxicity of Chromobacterium subtsugae strain

PRAA4

- 1 T to younger larval stages of bees. Because of t hese deficiencies, the study is unacceptable and cannot be used in ecological risk assessment for this active ingredient.

iii) Conclusions Regarding Nontarget Insects and Honey Bees

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

, on the MBI

- 203 EP Bioinsecticide

, MBI

- 203 SC, and Grandevo TM label s :

Environmental Hazards Statements

This product is highly toxic to bees exposed to direct treatment or residues on blooming crops or weeds. Do not apply this product or allow it to drift to blooming crops or weeds if bees are visiting the treatm ent area.

This product is toxic to certain nontarget terrestrial arthropods. Minimize spray drift away from target area to reduce effects to nontarget insects.Chromobacterium subtsugae strain PRAA4

- 1 T Page 17 of 41 Biopesticides Registration Act ion Document

Nontarget Plants (OCSPP Guideline 885.4300)

i) Data Submitted with Registration Applications for MBI

- 203 TGAI and MBI

- 203 EP

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 s oils and is not related to any known plant pathogen, effects of mild phytotoxicity were observed in cucumbers at 5X and 10X the field application rate, and in chrysanthemum flowers at all application rates.

ii) Data Submitted with Registration Application fo r MBI

- 203 SC

MRID No. 483335

- 02 : A greenhouse study was conducted to evaluate the phytotoxic effects of a single spray application of MBI

- 203 SC (active ingredient, 86.50% Chromobacterium subtsugae
strain PRAA4

- 1 T cells and spent fermentation media) on a variety of young potted monocot and dicot nontarget crop plants. The test included four treatments: the test material at application rates of 1 gallon/acre (the label recommended maximum application rate) or 3.3 gallon/acre, a positive control of Roundup ® at 1.4 gallon/acre, and a negative control of deionized water only. A phytotoxicity rating scale was used to rate adverse effects (e.g., necrosis, leaf curl/cup, tip burn, yellowing, stunting, chlorosis) at 2 to 8 days after treatment. The test material wa s applied to the plants tested at the field application rates of 1 gallon/acre or 3.3 gallon/acre, phyto to xicity symptoms ranged from none to mild. Squash, pea, and chrysanthemum appeared to have the worst signs of toxicity/pathogenicity, but effects were also seen in wheat, barley, onion, soybean, broccoli, bean, and tomato. Response in the positive control treatment was appropriate. The study is deficient in that it was not conducted until the plants were harvested, and no attempt was made
to reisolate th e microbial pest control agent from the plants; however, it does provide information useful for ecological risk assessment and is classified as supplemental.

iii) Conclusions Regarding Nontarget Plants

Given the effects noted in the nontarget plant studies, t he MBI

- 203 EP Bioinsecticide , MBI

- 203 SC, and Grandevo TM label s will require all users to test for phytotoxicity prior to pesticide application.

b. Aquatic Animals and Plants

Freshwater Fish ( Harmonized Guideline 885.4200), Freshwater Invertebrates ( Harmonized Guideline 885.4240), and Estuarine/Marine Fis h and Invertebrates ( Harmonized 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 d emonstrated 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

ra tionale 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 studie s (MRID No s

. 479450

- 12 and 481129

- 04) submitted by the applicant that tested the effects ofChromobacterium subtsugae strain PRAA4

- 1 T Page 18 of 41 Biopesticides Registration Act ion Document

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 Bioinsecticide is not intended for direct application into the estuarine or marine environment or expected to enter such environment i n significant concentrations.

Even though MBI

- 203 EP Bioinsecticide , MBI

- 203 SC, and Grandevo TM are not applied directly to aquatic environments, EPA is requiring set maximum labeled application rate s and specific
aerial drift reduction text , as well as certain mitigation language

, be included on the label s to reduce the potential for exposure to a quatic invertebrates . This information is presented in the
table below.

Table 1. Chromobacterium subtsugae PRAA4

- 1 T Label Text Required to Reduce Exposure to Aquatic Invertebrates

Product Name (EPA File Symbol or EPA

Registration Number) Maximum Applicat ion Rate Environmental Hazards Statements Buffer
Size 1 Droplet
Size 5

MBI

- 203 EP Bioinsecticide (84059

- 10) 12 quarts of product per acre This product is toxic to aquatic invertebrates. Drift or runoff may be hazardous to aquatic organisms in water adjacent to treated areas. 75 feet 2

Medium to
Coarse MBI

- 203 SC (84059

- RA) 60 feet 3

GrandevoTM (84059

- RT) Aerial : 3 pounds of product

per acre

Low Boom : 20 pounds of product per acre 0 feet 4

1 Buffer statement (in the Directions for Use) reads as follows: “Do not apply within XX feet of aquatic habitats (such as, but not limited to, lakes, reservoirs, rivers, streams, marshes, ponds, estuaries, and commercial fish ponds).”

2 See U.S. EPA 2011c and U.S. EPA 2011d for analysis.

3 See U.S. EPA 2012c for analy sis.

4 See U.S. EPA 2012d for analysis.

5 Droplet size (in the Directions for Use) reads as follows: “Use only medium or coarser spray nozzles according to ASAE (S572) definition
for standard nozzles.”

c. Open Literature Information

Literature searche s were performed to determine if other effects have been reported that were not contained within the information provided by the applicant. Both the Science Citation Index Expanded database (1970

- present) and the Environmental Information Search service (1 964

- 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; Na tional Technical Information Service; and Waternet databases. None of the records found in any of the searches described here reported effects on any nontarget species. The only results relevant to the active ingredient were two documents published by the United States Department of Agriculture

– Agricultural Research Service concerning the toxicity of Chromobacterium subtsugae to Colorado 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 (M artin et al . 2007b) . Neither document provided additional information on environmental or nontarget effects useful for the purposes of this risk assessment.Chromobacterium subtsugae strain PRAA4

- 1 T Page 19 of 41 Biopesticides Registration Act ion Document

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., Tiers II, III, and IV) is not req uired.

3

. Threatened and Endangered Species Assessment

EPA has determined that there is a potential for effects to nontarget organisms, including endangered and threatened species as a result of the proposed labeled applications (aerial sprays
and che migation) 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 specific effects and affected specie s. For this reason, aerial applications were not recommended. Terrestrial applications through sprinkler irrigation systems (chemigation) are expected to result in exposure to nontarget insects, though to a lesser extent than aerial applications. The end

- u se pesticide products are also labeled for greenhouse use, which will not result in significant exposure to nontarget organisms. Aquatic applications are not permitted for these pesticide products , which will limit exposure to aquatic invertebra tes and
oth er aquatic organisms. Mitigation language (e.g., requirement for a buffer between aquatic and

treatment areas , if needed, and use of only medium to coarser spray nozzles for aerial applications), addressing concerns of Chromobacterium subtsugae strain PRAA 4

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

- use pesticide product labels

.

The Biopesticides and Pollution Prevention Division (BPPD) makes “no effect” determinations for direct effects, ind irect 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 applications to the proposed crop plants treated within enclosed greenhouses

BPPD concludes that the following types of labeled applications of Chromobacterium subtsugae
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 likely to occur and the species potentially affected.

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

V. ENVIRONMENTAL JUSTICE

EPA seeks to achieve environmental justice

— the fair treatment and meaningful involvement of all people regardless of race, color, national origin, or income

— with respect to the development, implement ation, 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 result ing from industrial,Chromobacterium subtsugae strain PRAA4

- 1 T Page 20 of 41 Biopesticides Registration Act ion Document

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 opportun ity 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 th e 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 issu es, EPA seeks information on any groups or segments of the population who, as a result their location, cultural practices, or other factors, may have atypical, unusually high exposure to Chromobacterium subtsugae strain PRAA4

- 1 T

, compared to the general po pulation. 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 MANAGEMENT DECISION

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

(1) Its composition is such as to warrant the proposed claims for it; (2) Its labeling and other material required to be submitted comply with the requirements of FIFRA; (3) It will perform its intended functi on without unreasonable adverse effects on the environment; AND (4) When used in accordance with widespread and commonly recognized practice, it will not generally cause unreasonable adverse effects on the
environment.

To satisfy crit erion 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, MBI

- 203 EP Bioinsecticide , MBI

- 203 SC, or Grandevo TM labels, and such products are not expected to cause unreasonable adverse effects on the environment when used according to the label instructions. Criterion 2 is satisfied by the current product labels, as well as the data and information presented in th is 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 Bioinsecticide , MBI

- 203 SC, and GrandevoTM (end

- use pesticide product s ) , in particular, are likely to provide protection against insect and mite pests as claimed, satisfying criterion 3. Criterion 4 is satisfied in that the Chromobacterium subtsugae
strain PRAA4

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

- 203 TGAI

, MBI

- 203 EP Bioinsecticide

, MBI

- 203 SC, and Grandevo TM, containing Chromobacterium subtsugae strain PRAA4

- 1 T as an 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 willChromobacterium subtsugae strain PRAA4

- 1 T Page 21 of 41 Biopesticides Registration Act ion Document

likely require that additional data, in conjunction with a registration amendment request, be
submitted

.

V I I

. ACTIONS REQUIRED O F THE REGISTRANT

A. Final Printed Labeling

Before releasing pesticide products containing Chromobacterium subtsugae s train 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, Marrone Bio Innovations must submit the following data within one year of this product’s registration:

(1) Analysis of Samples ( Harmonized Guideline 885.1400) : A five

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

(2) Storage Stability ( Harmon ized Guideline 830.6317) and Corrosion Characteristics ( Harmonized Guideline 830.6320) : The results of a two

- year storage stability and corrosion characteristics study (ongoing).

As terms of the MBI

- 203 SC and Grandevo TM registrations, Marrone Bio Innovations must
submit the results of two

- year storage stability (Harmonized Guideline 830.6317) and corrosion characteristics studies (Harmonized Guideline 830.6320).

C. Reporting of Adverse Effects and Hypersensitivity Incidents

Notwithstanding the information stated in the previous sections, it should be clearly understood that certain, specific data are required to be reported to EPA as a requirement for maintaining the federal registration for a pesticide product. A brief summary of these types of data are described
below.

Reports of all incidents of adverse effects to the environment must be submitted to EPA under the provisions stated in FIFRA section 6(a)(2). Additionally, all incidents of hypersensitivity (includ ing 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 22 of 41 Biopesticides Registration Act ion Document

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

ASAE American Society of Agricultural Engineers

BPPD Biopesticides and Pollution Preven tion 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
CLPU Cabbage Looper Potency Unit(s)
CLPU/L Cabbage Looper Potency Units per liter CLPU/mg Cabbage Looper Potency Units per milligram

cP centipoise
EDSP Endocrine Disruptor Screening Program

EP end

- use product

EPA Environmental Protection Agency (the “Agency”)
FFDCA Federal Food, Drug, and Cosmetic Act

FIFR A Federal Insecticide, Fungicide, and Rodenticide Act FQPA Food Quality Protection Act

FR Federal Register g/ mL gram s per milliliter
LC 50 median lethal concentration. A statistically derived concentration of a substance that can be expected to cause dea th 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).

μg microgram (s) μL microliter (s) mg/kg milligram s per kilogram mg/L milli gram s per liter

mL milliliter (s)

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 23 of 41 Biopesticides Registration Act ion Document

IX. BIBLIOGRAPHY

A. Stud ies Submitted to Support the Chromobacterium subtsugae strain PRAA4

- 1 T

Pesticide Product Registrations

MRID No. Study Information

479450

- 01 Koivunen M. 2009

. Product Chemistry for MBI

- 203 TGAI . Unpubli shed 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 Toxici ty 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

– Final Report. Project N umber: 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
Stillme adow, 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 Sensitization 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 . Unpublished 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 a ges

.

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 24 of 41 Biopesticides Registration Act ion Document

MRID No. Study Information

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, Li mited, 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 Gallagh er 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. Proje ct Number: 2702/145 . Unpublished study prepared by Marrone 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 Innova tions, 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,
Li mited, 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

.

479 450

- 20 Richards K. 2009. Evaluation of the Dietary Effect(s) of MBI 203 on Adult Honey Bees ( Apis mellifera L.)

– Final Report. Project Number: CAR/140/09B. Unpublished study prepared by California Agricultural Research, Incorporated, 12

pages.

479450

- 21 Richards K. 2009. Evaluation of the Dietary Effect(s) of MBI 203 on Adult Honey Bees ( Apis mellifera L.)

– Final Report. Project Number: CAR/140/09A. Unpublished study prepared by California Agricultural Research, Incorporated, 11

pages.

479450

- 22 Miles on 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 25 of 41 Biopesticides Registration Act ion Document

MRID No. Study Information

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 stud y prepared by Technology Sciences Group, Incorporated, 77 pages.

479450

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

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

.

481129

- 02 Koivunen M

. 2010. Suppleme ntal Product Identity Study, Secondary Metabolites (MBI

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

.

481129

- 03 Chanbusarakum L

. 2010. Supplemental Manufacturing Process Study, Cabbage Looper Potency Unit Justification (MBI

- 203)

. Unpublished study prepared by Marrone Bio Innovations, 25 pages.

481129

- 04 Gallagher S, Krueger H. 2009. MBI

- 203: A 21

- Day Life Cycle Toxicity and Pathogenicity Test with the Cladoceran ( Daphnia magna )

. Project Number:
664A/101. Unpublished study pr epared 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, Inco rporated, 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.

481129

- 08 Pitts K

. 2010. Response to Tier 1 Microbial Pesticide Data Requirements for

MBI

- 2 03 EP: Supplemental Study for Revised EP Formulation. Unpublished study prepared by Marrone Bio Innovations, 1 23 pages.

481307

- 01 Chanbusarakum L

. 2010. MBI 203 TGAI: Supplemental Product Identity Study,
Mode of Action. Unpublished study prepared by M arrone Bio Innovations, 30

pages.

482844

- 01 Younger C. 2010. MBI

- 203: Honey Bee Acute Contact Toxicity Limit Test

– Amended Final Report. Project Number: 14300/10. Unpublished study prepared by Stillmeadow, Inc., 13 pages.Chromobacterium subtsugae strain PRAA4

- 1 T Page 26 of 41 Biopesticides Registration Act ion Document

MRID No. Study Information

482844

- 02 Younger C. 2010. MBI

- 203: Ladybird Beetle Non

- Target Insect Microbial Testing

– Final Report. Project Number: 14356/10. Unpublished study prepared by Stillmeadow, Inc., 14 pages.

483335

- 01 Koivunen M. 2010. Product Chemistry for MBI

- 203 SC. Unpublished study prepared by Marrone Bio Innovations, 93 pages.

483335

- 02 Lewis S. 2010. MBI

- 203 SC: Nontarget Plants. Project Number: 5302/63, 5302/66, 5302/76. Unpublished study prepared by Marrone Bio Innovatio ns, 14

pages.

483335

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

MBI

- 203 SV: Supplemental Study for Revised EP Formulation. Unpublished study prepared by Marrone Bio Innovations, 79 pages.

483335

- 04 Younger C. 2010. MBI

- 203 TGAI: Honey Bee Larval Non

- Target Topical/Oral Toxicity Microbial Test

– Final Report. Project Number: 14390/10. Unpublished study prepared by Stillmeadow, Inc., 12 pages.

484406

- 01 Pitts K

. 2011. MBI 203 TGAI: 12

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

21 p ages

.

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

.

486220

- 01 Pitts K. 2011. Product Chemistry for MBI

- 203 DF1: (Group A & B). Project Number: MBI/203/DF1. Unpublished study prepared by Marrone Bio Innovations, 70 pages.

486220

- 02 Pitts K. 2011. Product Chemistry for MBI

- 203 DF2: (Group A & B ). Project Number: SOP/MBI/G5/2009/V2. Unpublished study prepared by Marrone Bio Innovations, 64 pages.

486220

- 03 Pitts K. 2011. Response to Tier 1 Microbial Pesticide Data Requirements for

MBI

- 203 DF1. Project Number: EMP76831. Unpublishe d study prepared by Marrone Bio Innovations, 53 pages.

486220

- 04 Pitts K. 2011. Response to Tier 1 Microbial Pesticide Data Requirements for

MBI

- 203 DF2. Unpublished study prepared by Marrone Bio Innovations, 61

pages.Chromobacterium subtsugae strain PRAA4

- 1 T Page 27 of 41 Biopesticides Registration Act ion Document

B. Environmenta l Protection Agency Risk Assessment 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 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 a nd Mites in Ornamental Plants, Turf and Edible Crops, Memorandum from A. Gross and Z. Vaituzis, Ph.D. to J. Kausch dated July 7, 2011.

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

- I. Analysis drafted by S. Borges and dat ed August 11, 2011.

U.S. EPA. 2011d. Spray Buffers Revised C

. subtsugae . Spreadsheet assembled by S. Borges and dated August 17, 2011.

U.S. EPA. 2012a. Chromobacterium subtsugae strain PRAA4

- 1 T

. Memorandum from J. Facey , Ph.D. and J.L. Kough, Ph. D. to J. Kausch dated January 5, 2012.

U.S. EPA. 2012b. Chromobacterium subtsugae strain PRAA4

- 1 T

. Memorandum from J.V. Gagliardi , Ph.D. and J.L. Kough, Ph.D. to J. Kausch dated January 19, 2012.

U.S. EPA. 2012 c . Review of Nonta r get Effects Studies wi th Chromobacterium subtsugae strain

PRAA4

- 1 T to Support the Section 3 Registration of a New Product, MBI

- 203 SC; PC Code 016329; EPA File Symbol 84059

- RA. Memorandum from S. Borges to J. Kausch dated January 23, 2012.

U.S. EPA. 2012d. Review of Environmen tal Hazards Statements and Spray Buffer Requirements
for a New End

- Use Product, GRANDEVOTM, Containing the Active Ingredient Chromobacterium subtsugae strain PRAA4

- 1 T ; PC Code 016329; EPA File Symbol
84059

- RT. Memorandum from S. Borges to J. Kausch dated Ma rch 8, 2012.

C. Other References

Aislabie J, Smith JJ, Fraser R, McLeod M. 2001. Leaching of bacterial indicators of faecal contamination through four New Zealand soils. Australian Journal of Soil Research
39:1397

- 1406.

Centers for Disease Contr ol and Prevention. 2009. Drinking Water

- Water Treatment. Available
from http://www.cdc.gov/healthywater/drinking/public/water_treatment.html

.

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

.

Lindow SE, Brandl MT. 2003. Microbiology of the phyllosphere. Applied and Environmental
Microbi ology 69:1875

– 1883.Chromobacterium subtsugae strain PRAA4

- 1 T Page 28 of 41 Biopesticides Registration Act ion Document

Martin PAW, Gundersen

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

sp. n ov., a betaproteobacterium toxic to Colorado potato beetle and other insect pests. International Journal of Systematic and Evolutionary Microbi ology 57:993

– 999.

Martin PAW, Hirose E, Aldrich JR. 2007b. Toxicity of Chromobacterium subtsugae to southern green stink bug (Heteroptera: Pentatomidae) and corn rootworm (Coleoptera: Chrysomelidae). Journal of Economic Entomology 100(3):680

– 684.

Pang L, 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 Journal 7:97

- 111.

U.S. EPA. 2004. Primer for Municipal Wastewater Treatment Systems. EPA 832

- R

- 04

- 001.
Available from http://www.epa.gov/npdes/pubs/primer.pdf

.

U.S. EPA. 1996. Microbial Pesticide Test Guidelines

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

.Chromobacterium subtsugae strain PRAA4

- 1 T Page 29 of 41 Biopesticides Registration Act ion 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 )

Harmonized

Guideline
Number Data Requ irement 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 medi a (minimum of 1,5 00 Cabbage Looper Potency (or Killing) Units per milligram). 479450

- 01
481129

- 02
481307

- 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 is on deposit with the Agricultural Research Service Culture Collection (also known as the Northern Regional Research Laboratory Collection) in Peoria, Ill inois under
Accession Number NRRL B

30655. Not applicable 479450

- 01

885.1300 Discussion of Formation
of Unintentional Ingredients Submitted data fulfill the requirement for discussion of formation of unintentional ingredients. 479450

- 01

885.1400 Analys is of Samples Submitted data fulfill the requirement for analysis of samples. 479450

- 01

885.1500 Certification of Limits Not applicable Limits listed on the confidential
statement of formula are adequate/acceptable. 479450

- 01

Additional Studies

830.1800 Enforcement Analytical
Method Not applicable Submitted data fulfill the requirement for an enforcement analytical method. 479450

- 01Chromobacterium subtsugae strain PRAA4

- 1 T Page 30 of 41 Biopesticides Registration Act ion Document

TABLE 2. Product Analysis Data Requirements for the End

- Use Product ( E P), MBI

- 203 EP Bioinsecticide (40 CF R § 158.2120)

Harmonized
Guideline Number Data Requirement Results MRID No.

885.1100 Product Identity Submitt ed data fulfill the requirement for product identity. 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
481 129

- 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 u nintentional 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 f ormula are adequate/acceptable. 479450

- 02

Additional Studies

830.1800 Enforcement Analytical
Method Submitted data fulfill the requirement for an enforcement analytical method. 479450

- 02

TABLE 3. Product Analysis Data Requirements for the End

- Use Product (EP), GrandevoTM (40 CFR § 158.2120)

Harmonized
Guideline Number Data Requirement Results MRID No.

885.1100 Product Identity Submitted data fulfill the requirement for product identity. GrandevoTM contains 30.0% 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). 486220

- 01
486220

- 02

885.1200 Manufacturing Process Submitted data fulfill the requirement for manufacturing

process. Not app licable 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 in gredients. 486220

- 01
486220

- 02

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

885.1500 Certification of Limits Limits listed on the confidential statement of formula are adequate/acceptable.Chromobacterium subtsugae strain PRAA4

- 1 T Page 31 of 41 Biopesticides Registration Act ion Document

TABLE 4 . Prod uct Analysis Data Requirements for the End

- Use Product (EP), MBI

- 203 SC (40 CFR § 158.2120)

Harmonized
Guideline Number Data Requirement Results MRID No.

885.1100 Product Identity Submitted data fulfill the requirement for product identity. MBI

- 203 SC c ontains 86.50 % 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). 483335

- 01

885.1200 Manufacturing Process Submitted data fulfill the requirem ent for manufacturing

process. 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 dis cussion of formation of unintentional ingredients. 483335

- 01

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

885.1500 Certification of Limits Limits listed on the confidential statement of formula are adequat e/acceptable.

TABLE 5 . Physical and Chemical Characteristics for the Technical Grade of the Active Ingredient (TGAI)/Manufacturing

Use Product (MP), MBI

- 203 TGAI (40 CFR § 158.2120)

Harmonized

Guideline
Number Data Requirement Results MRID No. TG AI MP

830.6302 Color Purple

- blue Not applicable 479450

- 01

830.6303 Physical State Liquid Not applicable 479450

- 01

830.6304 Odor Grass

- like Not applicable 479450

- 01

830.6313 Stability to Normal and
Elevated Temperatures, Metals,
and Metal Ions Expect ed 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 p esticide (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 32 of 41 Biopesticides Registration Act ion Document

TABLE 6 . Physical and Chemical Characteristics for the End

- Use Product s (EP s ), MBI

- 203 EP Bioin secticide and GrandevoTM (40 CFR § 158.2120)

Harmonized
Guideline Number Data Requirement Results
MRID No. MBI

- 203 EP
Bioinsecticide * GrandevoTM **

830.6302 1 Color Purple

- blue Basic Formulation : Light Purple
Alternate Formulation : Dark

Brown 479450

- 02*
486220

- 01**
486220

- 02**

830.6303 1 Physical State Liquid Powder 479450

- 02*
486220

- 01**
486220

- 02**

830.6304 1 Odor Herbaceous Fruity 479450

- 02*
486220

- 01**
486220

- 02**

830.6313 Stability to Normal and Elevated Temperatures, Metals, and Metal Ions Not appl icable Not applicable

830.6317 Storage Stability Expected to be stable for two years when stored per label
instructions. As term s of the MBI

- 203 EP Bioinsecticide and
Grandevo TM registration s , 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*
486220

- 01**
486220

- 02**

830.6319 Miscibility Miscible with water 479450

- 02*
486 220

- 01**
486220

- 02**

830.6320 Corrosion
Characteristics Not expected to be corrosive to packaging materials. As terms of

the MBI

- 203 EP Bioinsecticide and GrandevoTM registrations, 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*
486220

- 01**
486220

- 02**

830.7000 1 pH 6.2 Basic Formulation : 4.78
Alternate Formulation : 7.95 479450

- 02*
486220

- 01**
486220

- 02**

830.7100 Viscosity 21.8 cP Not required be cause the end

- use pesticide product, G randevoTM, is not a liquid form of microbial pesticide (refer to test note #4 of 40 CFR § 158.2120(d)). 479450

- 02*

830.7300 1 Density/Relative Density/Bulk Density

(Specific Gravity) 1.02 g/mL Basic Formulation : 0.407 g/mL
Alternate Formulation : 0.36 g/mL 4 79450

- 02*
486220

- 01**
486220

- 02**

1 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
Bioinse cticide and/or Grandevo TM

, it is summarized appropriately in this table

.Chromobacterium subtsugae strain PRAA4

- 1 T Page 33 of 41 Biopesticides Registration Act ion Document

TABLE 7. Physical and Chemical Characteristics for the End

- Use Product (EP), MBI

- 203 SC (40 CFR § 158.2120)

Harmonized

Guideline
Number Data Requirement Results MRID No.

830.6302 1 Color Purple

- blue 48 3335

- 01

830.6303 1 Physical State Liquid

830.6304 1 Odor Herbaceous

830.6313 Stability to Normal and Elevated Temperatures, Metals,
and Metal Ions Not applicable Not applicable

830.6317 Storage Stability Expected to be stable for two years when stored per label
instructions. As a term of the MBI

- 203 SC registration, EPA is requiring the results of a two

- year storage stability and corrosion characteristics study (ongoing) be submitted within one year.

483335

- 01 830.6319 Miscibility Miscible with water

830.6320 Corrosion Characteristics Not expected to be corrosive to packaging materials. As
a term of the MBI

- 203 SC registration, EPA is requiring the results of a two

- year storage stability and corrosion characteristics study (ongoing) be
submitted with in one year.

830.7000 1 pH 5.91

830.7100 Viscosity 154.3 cP

830.7300 1 Density/Relative Density/Bulk Density (Specific Gravity) 1.05 g/mL

1 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 SC, it is summarized appropriately in this table.Chromobacterium subtsugae strain PRAA4

- 1 T Page 34 of 41 Biopesticides Registration Act ion Document

T ABLE 8

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

- Use Product (MP)
, MBI

- 203 TGAI (40 CFR § 158.2140)

Harmonized 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
479450

- 03. Classification: Acceptable Not applicable 479450

- 23

885.3150 Acute Pulmonary Toxicity/Pathogenicity Waived based on the results of
MRID Nos. 479450

- 11 and
479450

- 05. Classification: Acceptable Not applicable 479450

- 23

885.3200 Acute Injection
Toxi city/Pathogenicity (Intravenous) Not 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 during research, development, or testing of the TGAI/MP, were reported by the applicant. Any future hypersensitivity incidents must be reported to EPA (refer t o 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 To xicity Not applicable Oral LD 50 female rats > 5,000 mg/kg Classification: Acceptable
TOXICITY CATEGORY IV 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 IV 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 IV 479450

- 05

870.2400 Acute Eye Irritation Not applicable MBI

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

- 06

870.2500 Primary Dermal
Irritation Not applicable MBI

- 203 TGAI was slightly irritating to the skin of rabbits. Classification: Acceptable
TOXICITY CATEGO RY IV 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 35 of 41 Biopesticides Registration Act ion Document

T ABLE 8

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

- Use Product (MP)
, MBI

- 203 TGAI (40 CFR § 158.2140)

Harmonized Guideline
N umber Data Requirement Results MRID N o. TGAI M P

Additional Studies

870.2600 Skin Sensitization Not a dermal sensitizer
Classifi cation: 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/Pathogenicity

(In travenous) 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 36 of 41 Biopesticides Registration Act ion Document

TABLE 9 . Toxicology Data Requirements for the End

- Use Product s (EP s ), MBI

- 203 EP Bioinsecticide and GrandevoTM (40 CFR § 158.2140)

Harmonized

Guideline
Number Data Requirement Results
MRID No. MBI

- 203 EP
Bioinsecticide* GrandevoTM**

885.3050 Acute Oral Toxicity/Pathogenicity Not applicable Not applicable

885.3150 Acut e 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 rea ctions of humans and domestic animals that
occurred during research, development, or testing of the EP s , 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 appl icable

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 not expected to be of toxicological concern (refer to test note #5 of 40 CFR § 158.2140(d)) and are cleared for food use. Classification: Acceptable
TOXICITY CATEGORY III 479450

- 24 *
481129

- 08 *
486220

- 03**
486220

- 04**

870.1200 Acute Dermal Toxicity Waived based on the results of MRID No. 479450

- 04 and
bec ause this formulation contains inert ingredients that are not expected to be of toxicological concern (refer to test note #5 of 40 CFR § 158.2140(d)) and are cleared for food use. Classification: Acceptable
TOXICITY CATEGORY III 479450

- 24 *
481129

- 08 *
48622 0

- 03**
486220

- 04**

870.1300 Acute Inhalation Toxicity Waived based on the results of MRID No. 479450

- 05 and because this formulation contains inert ingredients that are not expected to be of toxicological concern (refer to test note #5 of 40 CFR § 158.21 40(d)) and are cleared for food use. Classification: Acceptable
TOXICITY CATEGORY III 479450

- 24 *
481129

- 08 *
486220

- 03**
486220

- 04**

870.2400 Acute Eye Irritation Waived based on the results of MRID No. 479450

- 06 and
because this formulation contains iner t ingredients that are not expected to be of toxicological concern (refer to test note #5 of 40 CFR § 158.2140(d)) and are cleared for food use. Classification: Acceptable
TOXICITY CATEGORY III 479450

- 24 *
481129

- 08 *
486220

- 03**
486220

- 04**

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 not expected to be of toxicological concern (refer to test note #5 of 40 CFR § 158.2140(d)) and are cleared for food use

. Classification: Acceptable
TOXICITY CATEGORY III 479450

- 24 *
481129

- 08 *
486220

- 03**
486220

- 04**Chromobacterium subtsugae strain PRAA4

- 1 T Page 37 of 41 Biopesticides Registration Act ion Document

TABLE 10. Toxicology Data Requirements for the End

- Use Product (EP), MBI

- 203 SC (40 CFR § 158.2140)

Harmonized

Guideline
Number Data Requirement Res ults MRID No.

885.3050 Acute Oral Toxicity/Pathogenicity Not applicable Not applicable

885.3150 Acute Pulmonary Toxicity/Pathogenicity Not applicable Not applicable

885.3200 Acute Injection Toxicity/Pathogenicity Not applicable Not applicable

885.3400 Hypersensitivity Incidents No hypersensitivity incidents, including immediate

- type or delayed

- type reactions of humans and domestic animals that occurred during research, development, or testing of the EP, were reported by the applicant. Any future hyperse nsitivity 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 thi s formulation contains inert ingredients that are not expected to be of toxicological concern (refer to test note #5 of 40 CFR § 158.2140(d)) and are cleared for food use. Classification: Acceptable
TOXICITY CATEGORY III

483335

- 03 870.1200 Acute Dermal To xicity Waived based on the results of MRID No. 479450

- 04 and because this formulation contains inert ingredients that are not expected to be of toxicological concern (refer to test note #5 of 40 CFR § 158.2140(d)) and are cleared for food use.
Classificati on: Acceptable
TOXICITY CATEGORY III

870.1300 Acute Inhalation Toxicity Waived based on the results of MRID No. 479450

- 05 and because this formulation contains inert ingredients that are not expected to be of toxicological concern (refer to test note #5 of 40 CFR § 158.2140(d)) and are cleared for food use. Classification: Acceptable
TOXICITY CATEGORY III

870.2400 Acute Eye Irritation Waived based on the results of MRID No. 479450

- 06 and because this formulation contains inert ingredients that are not

e xpected to be of toxicological concern (refer to test note #5 of 40 CFR § 158.2140(d)) and are cleared for food use. Classification: Acceptable
TOXICITY CATEGORY III

870.2500 Primary Dermal Irritation Waived based on the results of MRID No. 479450

- 07 and because this formulation contains inert ingredients that are not expected to be of toxicological concern (refer to test note #5 of 40 CFR § 158.2140(d)) and are cleared for food use. Classification: Acceptable
TOXICITY CATEGORY IIIChromobacterium subtsugae strain PRAA4

- 1 T Page 38 of 41 Biopesticides Registration Act ion Document

TABLE 11 . Non target 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)

Harmonized
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 provide s 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 haza rds to wild mammals. Testing indicates no adverse effects to laboratory rats when dosed orally at 5,000 mg MBI

- 203 TGAI/kg body weight. Classification: Acceptable 479450

- 03

885.4200 Freshwater Fish Toxicity/Pathogenicity The study indicates no adverse eff ects 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 , a nd 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 end

- use pestici de product
label s : “This product is toxic to aquatic invertebrates. Drift and runoff may be hazardous to aquatic organisms in water adjacent to treated areas.” Additionally, the end

- use pesticide products will (1) bear a maximum application rate (see Table 1 on page 18) , (2 )
include a restriction that aerial a pplications not occur within a specified distance of aquatic habitats (see Table 1 on page 18)

, (3) require aer ial applicators to use medium or coarser spray nozzles, and ( 4 ) list certain requirements and instructions to
further reduce potential for spray drift to offsite areas

. 885.4280 Estuarine/Marine Fish and Invertebrate Testing Not required because Chromobacterium subtsugae strain

PRAA4

- l T will not be applied to water and is not expected to

ente r 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 i n 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
483335

- 02

The test material was applied to the plants tested at the field application rates of 1 gallon/acre or 3.3 gallons/acre, and phytotoxicity symptoms ranged from none to mild. Squash, pea, and chrysanthemum appeared to have the worst signs of toxicity/pathogenicity, but effects were also seen in wheat, barley,
onion , soybean, broccoli, bean, and tomato. Classification: Supplemental but acceptable for risk

assessment Given the effects noted in the nontarget plant studies, the MBI

203 EP Bioinsecticide, MBI

- 203 SC, and Grandevo TM labels will require all users to test for phytotoxicity prior to pesticide application.Chromobacterium subtsugae strain PRAA4

- 1 T Page 39 of 41 Biopesticides Registration Act ion Document

TABLE 11 . Non target 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)

Harmonized
Guideline Number Data Requirement Results MRID No.

885.4340 Nontarget Insect Testing Tests indicate toxicity to ladybird beetles. There were no observed effects to green lacewing larvae or parasitic wasps. The
dosin g and duration of the parasitic wasp study, however, were
insufficient for a conclusive determination of no effects. Classification: Supplemental but acceptable for risk

assessment 479450

- 16
479450

- 17
479450

- 18
481129

- 05
482844

- 02

The dietary no

- obser v ed

- effect concentration of MBI

- 203 TGAI was considered to be greater than 10X the field application rate of 4 quarts/acre; however, it is not possible to confirm this dose. It is likely that the dose was at least equal to the expected
environmental concent ration for nontarget insects at the lowest application rate. Classification: Supplemental but acceptable for risk

assessment The following mitigating language is required in the
Environmental Hazards section of the end

- use pesticide product
label s : “Th is product is toxic to certain nontarget terrestrial arthropods. Minimize spray drift away from target area to reduce effects to nontarget insects.” Further testing would be required for the removal of this mitigating language from the end

- use pesticide pr oduct label s . Additionally, the end

- use pesticide product label s 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

. Classif ication: Supplemental but acceptable for risk

assessment 479450

- 19
479450

- 20
479450

- 21
482844

- 01
483335

- 04 The 48

- hour contact LD 50 for MBI

- 203 TGAI was greater than 25 μg active ingredient/bee. Classification: Supplemental because not of sufficient duration to determine pathogenicity and testing was not performed at a high dose. There were no adverse physiological responses noted in the test material or control grou p bees, but the study had several
deficiencies (e.g., information about the conduct of the experiment was lacking and different larval stages were used for different treatments)

. Classification: Unacceptable and cannot be used for risk

assessment
The f ollowing mitigating language is required in the
Environmental Hazards section of the end

- use pesticide product
label s : “This product is highly toxic to bees exposed to direct treatment or residues on blooming crops or weeds. Do not apply this product or al low 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 end

- use pesticide product label s . Additionally, the end

- use pesticide product label s will list certain requirements 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 40 of 41 Biopesticides Registration Act ion Document

TABLE 11 . Non target 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)

Harmonized
Guideline Number Data Requirement Results MRID No.

Addit ional Studies Not applicable Endangered Species
Evaluation Sufficient information was provided to determine that any potential risk from use of MBI

- 203 TGAI to endangered mammalian, avian, or fish species would likely be below the level of concern, and th at adverse effects to endangered plant species are unlikely. Sufficient information was not provided to determine the potential risk to endangered nontarget insects or aquatic invertebrates. BPPD will make an independent endangered species determination.
C lassification: Supplemental

– not a required study 479450

- 22Chromobacterium subtsugae strain PRAA4

- 1 T Page 41 of 41 Biopesticides Registration Act ion Document

APPENDIX B. PESTICIDE PRODUCTS

EPA Registration
Number /File Symbol Registration
Name Percentage
Active Ingredient Formulation Type Use Site(s) Method(s) of Application Application
Rate Target Pest

84059

- 9 MBI

- 203
TGAI 100.00% Technical N/A N/A N/A N/A

84059

- 10 MBI

- 203 EP
Bioinsecticide 94.50% End Use

– Liquid Various agricultural and greenhouse crops

(e.g., vegetables, ornamentals, and turf) Standard ground or aerial spray application equipment and

chemigation 4

– 12 quarts

per acre

Various insects
and mites 84059

- RA MBI

- 203 SC 86.50% End Use

– Liquid

84059

- RT GrandevoTM 30.0% End Use

– Powder Various agricultural and greenhouse crops,

as well as plants in residential settings (e.g., vegetables, ornamentals, and turf) Variable (see
draft label in

docket for specifics)