BIOPESTICIDES REGISTRATION ACTION DOCUMENT



## Killed, nonviable Streptomyces acidiscabies strain RL-110 T



## Pesticide Chemical (PC) Code: 016328

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

Revised April 22, 2012Killed, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 2 of 32

TABLE OF CONTENTS

I. EXECUT IVE SU MMARY ........................................................................................................ ................... 4

II. ACTIVE INGRED IENT OVERVIEW .............................................................................................. ......... 5

III. REGULATO RY BACKGR OUND .................................................................................................. ............ 5

A. Applications for Pe sticide Regi strati on ................................................................................... ........ 5

B. Food Tolera nce Exem ptio n .................................................................................................. ............ 5

IV. RISK A SSESSMEN T ............................................................................................................. ..................... 6

A. Product Analysis Asse ssment (40 CFR § 158.2120) ....................................................................... 6

B. Human Health Risk A ssessment (40 CF R 158.2140) ..................................................................... 7

C. Environmental Assessm ent (40 CFR § 158.2150) ........................................................................ 14

V. ENVIRONMENTA L JUST ICE ......................................................................................................... ........ 19

VI. RISK MANAGEMEN T DECI SION....................................................................................................... ... 20

VII. ACTIONS REQUIRED OF THE REGI STRANT ................................................................................... 21

A. Final Prin ted Labe ling .................................................................................................... ............... 21

B. Terms of Registra tion

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

C. Reporting of Adverse Effects and Hypersensitivi ty Incide nts .................................................... 21

VIII. BIBLIOGRAP HY .................................................................................................................. ..................... 21

A. Studies Submitted to Support the Registration of Pesticide Pr oducts Containing Killed, Nonviable Streptomyces acidiscabies strain RL-110 T ........................................................ 21

B. Environmental Protection Agen cy Risk Assessmen t Memora nda ............................................. 24

C. Other Referen ces .......................................................................................................... .................. 24

APPENDIX A. MICROBIAL PESTIC IDES DATA REQU IREMENTS ............................................................ 25

APPENDIX B. PEST ICIDE PR ODUCTS ............................................................................................... ............... 32Killed, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 3 of 32

BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

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

Science Review

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

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

Environmental Effects

Regulatory Review

Ann Sibold Regulatory Action Leader Alan Reynolds Team Leader, Microbial Pesticide Branch (MPB) Sheryl K. Reilly, Ph.D. Chief, MPBKilled, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 4 of 32

I. EXECUTIVE SUMMARY

Description of the Active Ingredient: Streptomyces acidiscabies strain RL-110 T is a naturally occurring bacterium that lives on dead organic matter (e.g., leaf litter). The killed, nonviable form of this microorganism is proposed for use as a pre- and post-emergent herbicide applied pre-plant, at-plant, and post-plant to control annual grasses, broadleaf and sedge weeds in landscape turf and ornamentals, residential lawns, and agricultural crops.

Assessing Risks to Human Health: Appropriate tests found no evidence that the living microorganism is toxic or pathogenic to human s, the microorganism is unable to grow at mammalian body temperatures, and the active ingredient contained in the proposed pesticide product is a killed, nonviable Streptomyces acidiscabies strain RL-110 T , so no harmful health effects to humans are expected from its use as a pesticide.

Assessing Risks to the Environment: P esticide products containing killed, nonviable Streptomyces acidiscabies strain RL-110 T are not likely to pose a risk to the environment or to any nontarget organisms.

Public Participation: On October 1, 2009, the U.S. Environmental Protection Agency (EPA, or Agency) announced a policy to provide a more meaningful opportunity for the public to participate in major registration decisions before they occur. According to this policy, EPA provides a public comment period prior to making a registration decisi on for 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 d ecisions more transparent, the proposed pesticide products (i.e., MBI-005 TGAI and MBI-005 EP) c ontaining a new active ingredient, killed,

nonviable Streptomyces acidiscabies strain RL-110 T , were subject to a 30-day comment period. In addition to containing a new active ingredient, th e registration of these pesticide products would result in the first outdoor, residential and food uses for killed, nonviable Streptomyces acidiscabies

strain RL-110 T

. The following documents were made available for comment in the docket, identification number EPA-HQ-OPP-2010-0079, and accessed through either http://www.regulations.gov or http://www.epa.gov/pesticides/regulating/registration-status.html : (1) draft Killed, Nonviable Streptomyces acidiscabies strain RL-110 T Biopesticides Registration Action Document (BRAD); (2) draft label for the manufacturing-use pesticide product, MBI-005TGAI (EPA File Symbol 84059-RR); and (3) draft label for the end-use pesticide product, MBI005 EP (EPA File Symbol 84059-RE). The public participation comment period was initiated on March 12, 2012. No comments were received during the comment period. Based upon the data and other information submitted in support of the proposed registrations, EPA is issuing these registrations under Section 3(c)(5) of the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA). The basis for this preliminary decision is characterized throughout this BRAD.

For definitions of scientific terms, please refer to http://www.epa.gov/pesticides/glossary/

.Killed, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 5 of 32
II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Killed, nonviable Streptomyces acidiscabies strain RL-110 T

Culture Collection: American Type Culture Collection (ATCC) in Manassas, Virginia

under ATCC 49003

OPP Chemical Code: 016328

Type of Pesticide: Microbial Pesticide – Herbicide

See Appendix B for specific information (e.g., use sites, application rates, methods of application, formulation types, and target pests) regarding the proposed pesticide products containing this active ingredient.

III. REGULATORY BACKGROUND

A. Applications for Pesticide Registration

On December 29, 2009, Marrone Bio Innovations, Inc. (MBI; 2121 Second Street, Suite B-107, Davis, CA 95618) submitted an application for re gistration of the manufacturing-use pesticide product, MBI-005 TGAI (EPA File Symbol 84059-RR), containing the new microbial active ingredient killed, nonviable Streptomyces acidiscabies strain RL-110 T . In the Federal Register (FR) of March 10, 2010 ( 75 FR 11175 ), EPA announced receipt of this application for a pesticide product containing a new active ingredient. Subsequent to the submission of the manufacturing-use pesticide product, on June 25, 2010, MBI submitted an application for registration of the end-use pesticide product, MBI-005 EP (EPA File Symbol 84059-RE), containing killed, nonviable Streptomyces acidiscabies strain RL-110 T for use as a herbicide on turf, cereal grains, corn and rice. In the Federal Register of November 24, 2010 ( 75 FR 71697 ), EPA announced receipt of an application for a pesticide product containing this new active ingredient as an herbicide for use on cereal grains, corn and rice. In the Federal Register of March 9, 2012, EPA announced receipt of an appli cation for registration of a pesticide product containing this new active ingredient for use on agricultural crops, residential lawns and landscape turf and ornamentals. No comments were received following publication of these notices. For more information, see the following docket at http://www.regulations.gov/: EPAHQ-OPP-2010-0079.

B. Food Tolerance Exemption

On December 29, 2009, MBI also submitted a petition that requested an exemption from the requirement of a tolerance for Streptomyces acidiscabies strain RL-110 T . In the Federal Register of March 10, 2010 ( 75 FR 11171 ), EPA announced MBI, Inc.’s filing of a pesticide petition that proposed establishing an exemption from the requirement of a tolerance exemption for residues

of Streptomyces acidiscabies strain RL-110 T in or on all agricultural commodities. No commentsKilled, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 6 of 32 were received following this publication. For more information, see the following docket at http://www.regulations.gov/ : EPA-HQ-OPP-2009-0078.

Two modifications have been made to the requested tolerance exemption. First, based upon the data and information available to the Agency, EPA is adding the qualifiers “killed” and “nonviable” before the microorganism’s taxonomic name and unique identifier. Use of these qualifiers is now consistent with the representati on of this active ingredient in other associated regulatory documents and should assist in preven ting confusion regarding its nomenclature in the future. Second, EPA is changing “in or on all agricultural commodities” to “in or on all food commodities” to align with the terminology the Agency currently uses when establishing tolerance exemptions for residues of other like active ingredients.

IV. RISK ASSESSMENT 1

A. Product Analysis Assessment ( 40 CFR § 158.2120 )

Microbial pesticide product analysis data requirements 1 include product chemistry and composition, analysis and certified limits, and physical and chemical characteristics 1 data. Product chemistry and composition data include information about the identity of the active ingredient, the manufacturing process, deposition of a sample in a nationally recognized culture collection, and discussion of formation of uninten tional ingredients. Analysis and certified limits data include information on analysis of samples and certification of limits. Physical and chemical characteristics data describe basic characteristics of the registered pesticide products, including color, physical state, odor, stability, miscibility, pH, corrosion characteristics, viscosity and density.

S treptomyces spp. are commonly found in agricultural settings (i.e., soils and decaying plant material) and are present on fresh produce of all kinds with no known adverse effects (Faucher et
al. 1992; Lambert et al. 1989; Loria et al. 2008; Loria et al. 2006; Wanner 2009). The Manual of Clinical Microbiology (9 th edition) states that the primary ecological niche for aerobic actinomycetes, such as Streptomyces acidiscabies strain RL-110 T , is likely decaying plant material. The Manual of Clinical Microbiology (9 th edition) further states that infections caused by Streptomyces spp. are infrequent and limited to species unrelated to acidiscabies and does not identify Streptomyces acidiscabies as clinically significant.

The active ingredient (a.i.) is identified as killed, nonviable Streptomyces acidiscabies strain RL110 T (ATCC 49003) cells and spent fermentation media. This a.i. was initially isolated from scab-infected potatoes collected in Maine and New York.

The herbicidal mode of action is exposure to and uptake of thaxtomin A, which is produced by the plant pathogen, Streptomyces acidiscabies strain RL-110 T . The herbicidal mode of action for

1 For information on microbial data requirements, refer to http://www.epa.gov/fedrgstr/EPAPEST/2007/October/Day-26/p20828.htm

.Killed, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 7 of 32 thaxtomin A is based on disruption of cell wall synthesis, with inhibition of cellulose biosynthesis being the main effect (Fett et al . 1992).

Refer to Table 1 in Appendix A for a summary of product chemistry and composition, analysis

and certified limits data. Refer to Table 2 in Appendix A for a summary of physical and

chemical characteristics data.

All product analysis data requirements required for registration of killed, nonviable Streptomyces

acidiscabies strain RL-110 T have been fulfilled.

B. Human Health Risk Assessment ( 40 CFR 158.2140 )

1. Toxicity

All toxicology data requirements for killed, nonviable Streptomyces acidiscabies strain RL-110 T

have been fulfilled. Acceptable Tier I mammalian toxicology data and information support the registration of the proposed pesticide products. Tier II and Tier III studies were not required. The results of the submitted studies are summarized below; for more information, refer to Table 3 in Appendix A

.

a. Acute Toxicity/Pathogenicity – Tier I

Acute Oral Toxicity/Pathogenicity – Rat (Harmonized Guideline 885.3050) and Acute Pulmonary Toxicity/Pathogenicity – Rat (Harmonized Guideline 885.3150); Master Record Identification Number (MRID No). 47946817) : The applicant provided the following rationale to support data waivers for acute toxicity/pathogenicity and acute pulmonary toxicity/pathogenicity testing: (1) the acute oral and acute inhalation t oxicity studies described below demonstrated that killed, nonviable Streptomyces acidiscabies strain RL-110 T is practically nontoxic to rats; (2) the acute injection toxicity toxicity/pathogenicity study described below demonstrated that live Streptomyces acidiscabies strain RL-110 T is not toxic, infective and/or pathogenic to rats; (3) a literature search found that this microorganism is not known to be infective to mammals; and (4) Streptomyces acidiscabies strain RL-110 T is naturally occurring in soil in the U.S.

EPA waived the required data given that Streptomyces acidiscabies strain RL-110 T is killed, pathogenicity and/or infectivity in mammals is not expected, and it would be unnecessary to conduct additional studies to evaluate toxicity.

Acute Injection Toxicity/Pathogenicity (Intravenous)

– Rat (Harmonized Guideline 885.3200; MRID No. 47946808) : Thirty young adult Sprague-Dawley rats (15/sex) were injected with live Streptomyces acidiscabies strain RL-110 T at a dose of 9.0 x 10 6 colony-forming units (CFUs) per animal and observed for signs of toxicity and pathogenicity up to 21 days. Ten animals (5/sex) were treated with autoclaved Streptomyces acidiscabies strain RL-110 T as controls, and ten animals (5/sex) were not treated. All animals survived, gained weight and appeared normal during the study. No observable abnormalities were noted in any animal at necropsy. There were no significant variations among organ weights. The test organism was not detected in the blood, brain, lungs, spleen, liver, kidneys, mesenteric lymph nodes and cecum contents of treatedKilled, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 8 of 32 animals. The test organism was not evaluated in the tissues/organs of animals treated with autoclaved test material or from the untreated control group. Live Streptomyces acidiscabies

strain RL-110 T was not toxic, infective, and/or pathogenic to rats when dosed intravenously at

9.0 x 10 6 CFU per animal. This study was rated acceptable.

Hypersensitivity Incidents (Harmonized Guideline 885.3400) : Hypersensitivity incidents, including immediate-type and delayed-type reac tions of humans and domestic animals, that occur during the testing of the technical grade of the active ingredient or manufacturing-use product, or are otherwise known to the applicant, must be reported if they occur (refer to test note #3 of 40 CFR § 158.2140(d)).

Cell Culture (Harmonized Guideline 885.3500) : This study is not required because killed,

nonviable Streptomyces acidiscabies strain RL-110 T is not a virus (refer to test note #4 of 40 CFR § 158.2140(d)).

Acute Oral Toxicity

– Rat (Harmonized Guideline 870.1100; MRID No. 47946802) : Three fasted female Sprague-Dawley rats were given a si ngle oral gavage dose of MOI 005 (containing killed, nonviable Streptomyces acidiscabies strain RL-110 T ) at a concentration of 5,000 milligrams per kilogram (mg/kg) body weight. The animals were observed for 14 days. All animals survived and appeared normal throughout the study. One animal lost weight during the second week; all other animals gained weight throughout the study. No observable abnormalities were found at necropsy. The weight loss in one animal was determined to not be significant, since the acute injection toxicity/pathogenicity study (MRID 47946808) conducted with the live

microbe showed live Streptomyces acidiscabies strain RL-110 T was not toxic, infective or pathogenic to rats. The acute median lethal dose (LD 50 ) for female rats was greater than 5,000 mg/kg. This study was rated acceptable, and MOI 005 was classified as Toxicity Category IV.

Acute Dermal Toxicity – Rat (Harmonized Guideline 870.1200; MRID No. 47946803) : Ten young adult Sprague-Dawley rats (5/sex) were dermally exposed to 5,050 mg/kg of MOI 005 (containing killed, nonviable Streptomyces acidiscabies strain RL-110 T ) for 24 hours on a clipped dorsal surface comprising approximately 10% of the body surface area. Following exposure, the animals were observed for 14 days. All animals survived, gained weight, appeared normal and had no signs of dermal irritation throughout the study. No observable abnormalities were found in any animal at necropsy. The acute LD 50 for combined (male and female) rats was greater than 5,050 mg/kg. This study was rated acceptable, and MOI 005 was classified as Toxicity Category IV.

Acute Inhalation Toxicity

–Rat (Harmonized Guideline 870.1300; MRID No. 47946804) : Ten young adult Sprague-Dawley rats (5/sex) were exposed nose-only to MOI 005 aerosol (containing killed, nonviable Streptomyces acidiscabies strain RL-110 T ) for 4 hours at a concentration of 2.21 milligrams per liter (mg/L). The animals were observed for 14 days. All animals survived the study. Decreased activity was noted from all animals upon removal from the chamber through day 1 with recovery by day 2. Piloerection was noted on all animals upon removal from the chamber with recovery by day 1. All animals gained weight during the first week, and five males and four females lost we ight during the second week. One female gained weight throughout the study. Red lungs were noted in the female that gained weight throughoutKilled, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 9 of 32 the study. All other animals were emaciated and had swollen and pink lungs at necropsy. The acute median lethal concentration for combined (male and female) rats was greater than 2.21 mg/L. This study was rated acceptable, and MOI 005 was classified as Toxicity Category IV.

Primary Eye Irritation – Rabbit (Harmonized Guideline 870.2400; MRID No. 47946805) : Three (1 male, 2 female) New Zealand White rabbits were exposed to 0.1 milliliters (mL) of killed,

nonviable Streptomyces acidiscabies strain RL-110 T instilled into the conjunctival sac of the right eye and were observed at 1, 24, 48, and 72 hours. Irritation was scored by the method of Draize and classified by the system of Kay and Calandra. No corneal opacity or iritis was noted throughout the study. Positive conjunctival irritation was noted in one rabbit at one hour after test material instillation with resolution by 24 hours. The maximum average score was 2.7 at one hour after test material instillation. This study was rated acceptable, and MOI 005 was minimally irritating to the eye and classified as Toxicity Category IV.

Primary Dermal Irritation – Rabbit (Harmonized Guideline 870.2500; MRID No. 47946806) : In this study, one male and two female New Zealand White rabbits were dermally exposed to 0.5 mL of undiluted MOI 005 (containing killed, nonviable Streptomyces acidiscabies strain RL110 T ) for 4 hours on an approximately 2.5 square centimeter (cm 2 ) area of the body surface. The animals were observed at 1, 24, 48, and 72 hours after patch removal. Irritation was scored by the method of Draize. No dermal irritation was noted on any animal during the study. The primary irritation index was 0.0. This study was rated acceptable, and MOI 005 was not dermally irritating and classified as Toxicity Category IV.

Skin Sensitization – Guinea Pig (Harmonized Guideline 870.2600; MRID No. 47946807) : Thirty four young adult Hartley guinea pigs (17/sex) were tested using the Buehler Method. After three weekly consecutive inductions, no test or naive control animals showed any dermal reaction after challenge. This study was rated acceptable, and MOI 005 was found to be not a dermal

sensitizer. c. Endocrine Disruptors

As required under section 408(p) of the Federal Food, Drug, and Cosmetic Act (FFDCA), EPA has developed the Endocrine Disruptor Screening Program (EDSP) to determine whether certain substances (including pesticide active and other ingredients) may have an effect in humans or wildlife similar to an effect produced by a “naturally occurring estrogen, or other such endocrine effects as the Administrator may designate.” The EDSP employs a two-tiered approach to making the statutorily required determinations. Tier 1 consists of a battery of 11 screening assays to identify the potential of a chemical substance to interact with the estrogen, androgen, or thyroid (E, A, or T) hormonal systems. Chemi cals 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 a ny, of the Tier 2 tests are necessary based on the available data. Tier 2 testing is designed to identify any adverse endocrine-related effects caused by the substance, and establish a quantitative relationship between the dose and the E, A,

or T effect.

Between October 2009 and February 2010, EPA issued test orders/data call-ins for the first group of 67 chemicals, which contains 58 pesticide active ingredients and 9 inert ingredients.Killed, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 10 of 32 This list of chemicals was selected based on the potential for human exposure through pathways such as food and water, residential activity, and certain post-application agricultural scenarios. This list should not be construed as a list of known or likely endocrine disruptors.

Killed, nonviable Streptomyces acidiscabies strain RL-110 T is not among the group of 58 pesticide active ingredients on the initial list to be screened under the EDSP. Under FFDCA section 408(p), EPA must screen all pesticide ch emicals. Accordingly, EPA anticipates issuing future EDSP orders/data call-ins for all pesticide active ingredients.

For further information on the status of the EDSP, the policies and procedures, the list of 67 chemicals, the test guidelines and the Tier 1 screening battery, please visit our website: http://www.epa.gov/endo/

.

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

Section 408(c)(2)(A)(i) of FFDCA allows EPA to establish an exemption from the requirement for a tolerance (the legal limit for a pesticide chemical residue in or on a food) only if EPA determines that the exemption is “safe.” Sec tion 408(c)(2)(A)(ii) of FFDCA defines “safe” to mean that “there is a reasonable certainty that no harm will result from aggregate exposure to the pesticide chemical residue, including all anticipated dietary exposures and all other exposures for which there is reliable information.” This includes exposure through drinking water and in residential settings but does not include occupational exposure. Pursuant to section 408(c)(2)(B) of FFDCA, in establishing or maintaining in e ffect an exemption from the requirement of a tolerance, EPA must take into account the fact ors 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 exemption 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....” Additionall y, section 408(b)(2)(D) of FFDCA requires that EPA consider “available information concerning the cumulative effects of [a particular pesticide’s] . . . residues and other substances that have a common mechanism of toxicity.”

EPA performs a number of analyses to determine the risks from aggregate exposure to pesticide residues. First, EPA determines the toxicity of pesticides. Second, EPA examines exposure to the pesticide through food, drinking water, and through other exposures that occur as a result of pesticide use in residential settings.

Consistent with section 408(b)(2)(D) of FFDCA, EP A 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 also considered available information concerning the variability of the sensitivities of major identifiable subgroups of consumers, including infants and children.

Based on the acute toxicity/pathogenicity data and information discussed previously and presented in Table 3 in Appendix A , the data required for a FFDCA risk assessment for killed,

nonviable Streptomyces acidiscabies strain RL-110 T have been fulfilled.Killed, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 11 of 32 a. Aggregate Exposure

In examining aggregate exposure, section 408 of FFDCA directs EPA to consider availableinformation 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, residential and other lawns and landscapes, or buildings (residential and other indoor uses).

Food Exposure and Risk Characterization : Killed, nonviable Streptomyces acidiscabies strain

RL-110 T will be applied as a herbicide to agricultural crops pre-plant, at-plant and post-plant, and may be applied up to the day of harvest; exposure to this active ingredient through food is possible, but is expected to be minimal for the following reasons: (1) the proposed pesticide products will be diluted prior to application; (2) pre-plant applications will occur 1-45 days before planting; (3) at-plant applications will be broadcast and incorporated into the soil mechanically or by rainfall or sprinkler irrigation; (4) post-plant applications for trees will be made as a broadcast or banded application to soil surface below established trees or between tree rows and incorporated into the soil by rainfall, irrigation or mechanical incorporation; (5) post plant lay-by and split application will be applied between rows and incorporated into the soil; (6) application to dry- seeded rice fields is followed by flooding or, for water seed rice, partially draining and re-flooding the fields; and (7) rainfall and sprinkler irrigation will further wash residues of the pesticide from the treated crops.

Following application, the killed, non-viable Streptomyces acidiscabies strain RL-110 T will naturally degrade due to consumption by other biological organisms, including bacteria and fungi (Doumbou et al . 1998).

In the unlikely event that any residues of the pesticide remain in or on consumed food, no adverse effects would be expected, based on the l ack of toxicity and pathogenicity demonstrated in the submitted studies (refer to section IV.B.1.a. of this BRAD).

Drinking Water Exposure and Risk Characterization : Exposure to residues of killed, nonviable Streptomyces acidiscabies strain RL-110 T in consumed drinking water is unlikely, since the majority of the proposed use patterns (ground and aerial) include measures to incorporate the herbicide into the soil; however, residues may appear at low levels in ground and surface water from these uses due to runoff or drainage from treated fields, or by spray drift. These residues will be minimized by natural degradation of the active ingredient by microbial activity (Doumbou et al . 1998). Furthermore since application of the product is concentrated in upper soil strata, movement through the soils would likely filter out any remaining product.

The proposed directions for applications to established turf in landscapes provide for dilution of the product prior to application, but do not include measures to incorporate the product. Since established turf constitutes significant ground cover, this, in itself, would be expected to reduce the potential runoff of the pesticide into surface water and percolation to ground water.

The proposed directions for applications to ornamentals in landscapes specify dilution prior to application and incorporation by irrigation or raking into the soil. These measures, along withKilled, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 12 of 32 natural degradation and incorporation of the product into upper soil strata, will reduce the potential for runoff into surface or ground water.

The proposed use in rice provides the greatest potential for residues of killed, nonviable Streptomyces acidiscabies strain RL-110 T to appear in ground and surface water, since application to rice fields is followed by flooding the treated fields. If, following application to rice fields, residues of Streptomyces acidiscabies strain RL-110 T are transferred to surface or ground waters that are intended for eventual human consumption, and subjected to sanitation (e.g., chlorination, pH adjustments, filtration, high temperatures) in drinking water treatment plants, the residues would likely be removed from the finished drinking water. In the unlikely event that any residues of the pesticide occur in drinking water even after being processed at a water treatment facility, no adverse effects would be expected, based on the lack of toxicity and pathogenicity demonstrated in the submitted studies (refer to section IV.B.1.a. of this BRAD).

Non-occupational, Residential Exposure and Risk Characterization : Given the natural

occurrence of Streptomyces acidiscabies in soil (Faucher et al . 1992; Loria et al . 2006), nonoccupational and residential exposure may already be occurring. Application of killed,

nonviable Streptomyces acidiscabies strain RL-110 T to established turf in residential and landscape settings will result in exposure via the dermal and inhalation routes. Any such exposures are expected to be minimal since the concentration of killed, nonviable Streptomyces

acidiscabies strain RL-110 T is diluted prior to application and the a.i. is not expected to persist, as described above. In the unlikely event that the proposed uses of the pesticide result in residential, non-occupational exposure, no adverse effects would be expected, based on the lack of toxicity and pathogenicity demonstrated in the submitted studies (refer to section IV.B.1.a. of this BRAD).

b

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

Section 408(b)(2)(D)(v) of FFDCA requires that, wh en considering whether to establish, modify, or revoke a tolerance exemption, EPA consider “available information concerning the cumulative effects of [a particular pesticide’s] . . . residues and other substances that have a common mechanism of toxicity.”

EPA has found killed, nonviable Streptomyces acidiscabies strain RL-110 T does not appear to produce a toxic metabolite similar to that found in other substances or produced by other microorganisms. For the purposes of the associated tolerance action, EPA has assumed that killed, nonviable Streptomyces acidiscabies strain RL-110 T does not have a common mechanism of toxicity with other substances, and thus, section 408(b)(2)(D)(v) does not apply. For information regarding EPA’s efforts to identify chemicals that may have a common mechanism of toxicity and to evaluate the cumulative effects of such chemicals, see EPA’s website at http://www.epa.gov/pesticides/cumulative

.

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

FFDCA section 408(b)(2)(C) provides that, in cons idering the establishment of a tolerance or tolerance exemption for a pesticide chemical residue, EPA shall assess the available informationKilled, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 13 of 32 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 a pply an additional tenfold (10X) margin of exposure (safety) for infants and children in the case of threshold effects to account for prenatal and postnatal toxicity and the completeness of the database on toxicity and exposure unless EPA determines that a different margin of exposure (safety) will be safe for infants and children. This additional margin of exposure (safety) is commonly referred to as the Food Quality Protection Act Safety Factor. In applying this provision, EPA e ither retains the default value of 10X or uses a different additional safety factor when reliabl e data available to EPA support the choice of a

different factor.

Based on its analyses of the submitted toxicity data (refer to section IV.B.1.a. and Table 3 in Appendix A ), EPA concludes that there are no threshold effects of concern to infants, children, or adults when killed, nonviable Streptomyces acidiscabies strain RL-110 T is used as labeled in accordance with good agricultural practices. As a result, EPA also concludes that no additional margin of exposure (safety) is necessary to protect infants and children and that not adding any additional margin of exposure (safety) will be safe for infants and children.

Based on the same data and EPA analysis presented directly above, the Agency is able to conclude that there is a reasonable certainty that no harm will result to the U.S. population, including infants and children, from aggregate exposure to the residues of killed, nonviable Streptomyces acidiscabies strain RL-110 T when it is used as labeled and in accordance with good agricultural practices as an herbicide. Such exposure includes all anticipated dietary exposures and all other exposures for which there is reliable information. EPA arrived at this conclusion because, considered collectively, the data available on killed, nonviable Streptomyces

acidiscabies strain RL-110 T do not demonstrate toxic potential to mammals, including infants

and children.

3. Occupational Exposure and Risk Characterization

The available data (e.g., lack of toxicity noted fo r oral, dermal, and inhalation routes of exposure to killed, nonviable pesticide available on Streptomyces acidiscabies strain RL-110 T ) do not demonstrate toxic potential to persons occupationally exposed to this microbial pest control agent. Standard personal protective equipment (PPE) on the label, including an N95 equivalent respirator, long-sleeve shirt, long pants, shoes, socks and waterproof gloves, further mitigates unintentional exposures to applicators.Killed, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 14 of 32

4. Human Health Risk Characterization

EPA considered human exposure to killed, nonviable Streptomyces acidiscabies strain RL-110 T
in light of the standard for registration in FIFR A and the relevant safety factors in FFDCA. A determination has been made that no unreasonable adverse effects to the U.S. population in general, and to infants and children in particular, will result when killed, nonviable Streptomyces

acidiscabies strain RL-110 T pesticide products are used in accordance with EPA-approved labeling.

C. Environmental Assessment ( 40 CFR § 158.2150 )

1. Ecological Hazards

Data and other information (e.g., scientific literature) submitted by the applicant to support requests to waive nontarget organism testing for killed, nonviable Streptomyces acidiscabies

strain RL-110 T are sufficient to fulfill the relevant microbial pesticide data requirements and for risk assessment purposes. Further testing of nontarget organisms at higher tier levels (i.e., Tiers II, III, and IV) is not required. EPA performed an environmental risk assessment based on data and other information provided by the applicant, and has determined that adverse effects to nontarget organisms are not anticipated from the proposed pesticidal uses of killed, nonviable Streptomyces acidiscabies strain RL-110 T . Moreover, EPA has made a “No Effect” determination for direct and indirect effects to listed species and their designated critical habitats resulting from these same proposed pesticidal uses.

For a summary of the generic data requirements described in sections IV(C)(1), refer to Table 4

in Appendix A

.

a. Terrestrial Animals and Plants

Avian Oral Toxicity (Harmonized Guideline 885.4050) (MRID Nos. 47946811): Fourteen-dayold mallard ducks ( Anas platyrhynchos ) were dosed with the test material containing killed,

nonviable Streptomyces acidiscabies strain RL-110 T (63 milligrams of active ingredient per kilogram of body weight (mg a.i. per kg bw)) once daily for 5 days and observed for an additional 25 days. A sterile filtrate control group and a negative control group were included in the test. One bird in the test material group was euthanized on day 23 due to low body weight and poor general condition, attributed to a feeder malfunction and pen mate aggression. There was no mortality in the other 2 groups. There were no treatment related signs of toxicity in any of the birds during the study. Gross necropsy of the survivors revealed no treatment-related differences between the test material group or the sterile filtrate control group compared to the negative control group. There was no evidence of test material multiplication (plaques, foci) in any bird. The No Observed Effect Level (NOEL) was determined to be greater than 63 mg a.i. per kg bw per day for five days. This study was acceptable.

Avian Inhalation Toxicity Pathogenicity (Harmonized Guideline 885.4100) (MRID Nos. 47946817): The applicant provided the following rationale to support a data waiver for Avian Inhalation Toxicity Pathogenicity testing: (1) killed, nonviable Streptomyces acidiscabies strainKilled, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 15 of 32

RL-110 T was not toxic or pathogenic in an avian oral toxicity study; (2) the a.i. in MBI-005 TGAI is a naturally occurring bacterium commonly found in soils and water; and (3) a literature search yielded no reports of adverse effects in avian species.

EPA considers the toxicity component of the acute avian inhalation toxicity/pathogenicity data requirement to be fulfilled by the acute avia n oral toxicity study (MRID No. 47946811). Given that the a.i. is killed, pathogenicity and/or infectivity in birds are not expected. It would be inappropriate and unnecessary to conduct further testing to evaluate these endpoints. Thus, EPA waived the pathogenicity component of the acute oral toxicity/pathogenicity data requirement.

Wild Mammals Toxicity/Pathogenicity (Harmonized Guideline 885.4150) (MRID No. 47946817): The applicant provided the following rationale to support a data waiver for Wild Mammals Toxicity/Pathogenicity testing: (1) killed, nonviable Streptomyces acidiscabies strain

RL-110 T was not toxic in acute exposure studies in rats; (2) Streptomyces acidiscabies strain RL110 T was not pathogenic in an intravenous injection study in rats; (3) Streptomyces acidiscabies

strain RL-110 T is a naturally occurring bacterium commonly found in soils; and (4) a literature search yielded no reports of adverse effects in humans or other mammals.

EPA considers the toxicity component of the acute oral toxicity/pathogenicity data requirement to be fulfilled by the acute oral, dermal and inhalation toxicity studies in rats (MRIDs 47946802, 47946803, 47946804). Given that the a.i. is killed, pathogenicity and/or infectivity in mammals are not expected. It would be inappropriate and unnecessary to conduct additional testing to evaluate these endpoints. Thus, EPA waived testing for the pathogenicity component of the acute oral and inhalation toxicity/pathogenicity data requirements.

Nontarget Insects (Harmonized Guideline 885.4340) (MRID Nos. 47946814, 47946815 and 48104505) : Two studies were conducted to determine the toxic effects of killed, nonviable Streptomyces acidiscabies strain RL-110 T to adult ladybird beetles ( Hippodamia convergens ) and green lacewing ( Chrysoperla rufilabris ) larvae. In the adult ladybird study, killed, nonviable Streptomyces acidiscabies strain RL-110 T was administered in diet at nominal concentrations of 510, 5,100, or 51,000 parts per million (ppm). The test included a negative control group. The test was terminated on day 17, when mortality in the untreated control group exceeded 20%. Cumulative mortality in the 510 ppm, 5,100 ppm, and 51,000 ppm test material groups was 24%, 25%, and 32%, respectively, compared to 21% in the untreated control group. Surviving beetles in all groups were normal in appearance and behavior throughout the test. The no observed effect level (NOEL) for 17 day diet exposure was 51,000 ppm. This study was acceptable.

In the green lacewing ( Chrysoperla rufilabris ) larvae study, the larvae were provided killed,

nonviable Streptomyces acidiscabies strain RL-110 T in nominal concentrations of 510, 5,100, or 51,000 ppm in a moth egg diet. The test included a negative control group fed moth egg diet treated with reverse osmosis water only. The test was terminated when mortality in the negative control group exceeded 20% on day 10. Mortality in the 510 ppm, 5,100 ppm, and 51,000 ppm test material groups was 30%, 30%, and 33%, respectively, compared to 22% in the untreated control. No pupation occurred in any group, and th ere were no signs of toxicity. The NOEL for 10 day diet exposure was 51,000 ppm. This study was acceptable.Killed, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 16 of 32 A third nontarget insect lab study was conducted with the end-use product MOI-005 EP (containing 17% killed, nonviable Streptomyces acidiscabies strain RL-110 T and spent fermentation media) to determine the toxicity of this product to the parasitic wasp ( Aphidius

matricariae ), dusty miller plants ( Centaurea cineraria ) were artificially infested with green peach aphids ( Myzus persicae ) and sprayed with the test material at 10X the field application rate. The test included an inactivated test material group, a test material sterile filtrate group, and an untreated control. The treated plants and aphids were infested with parasitic wasps for 24 hours and observed for 22 days afterward. Beginning on day 8, plant leaves with mummies (parasitized aphids) were removed, placed in Petri dishes, and monitored for adult wasp emergence. At test end, emergence of adult wasps in the test material, inactivated test material, and sterile filtrate groups was 182%, 147%, and 100%, respectively, compared to 162% in the untreated control. Emergence exceeded 100% due to uncounted parasitized, but not yet mummified, aphids on the leaves that mummified after being placed in the Petri dishes. The test material did not appear to be toxic to the parasitic wasp when applied to plant hosts at 10X the field application rate. The EC 50 was greater than 11.04 mg/L; the NOEL was greater than 11.04 mg/L. This study was acceptable.

Honey Bees (Harmonized Guideline 885.4380) (MRID No. 47946816) : To determine the effects of killed, nonviable Streptomyces acidiscabies strain RL-110 T in the diet to adult honeybees ( Apis mellifera ), bees were provided a nominal test material concentration of 500 μg per mL in 30% sucrose solution for 20 days. A positive control and untreated control were included in the test. All bees appeared normal during the test. At test end, mortality in the test material group was 16.5%, compared to 30.0% in the untreated control group. The response of the positive control group was appropriate. There were no adverse effects of killed, nonviable Streptomyces

acidiscabies strain RL-110 T on adult honeybees under the conditions of this study. The NOEL was 500 μg per mL in 30% sucrose solution for 20 days. This study was acceptable.

Nontarget Plants (Harmonized Guideline 885.4300); (MRID No.47990702) : Bioassays were conducted to determine the effects of the end-use product MOI-005 EP containing 17% killed,

nonviable Streptomyces acidiscabies strain RL-110 T and spent fermentation media on nontarget crops. Potted plants were sprayed with the test ma terial at 2X, 1X, or 0.5X the field application rate. MBI-005 EP was phytotoxic to all dicots tested, even at the 0.5X rate. Phytotoxicity was mild to moderate on broccoli, tomato, and pea; moderate on lettuce and bean; and moderate to severe on soybean and cucumber. For monocots, no signs of phytotoxicity occurred on bluegrass, bentgrass, or rice. At the highest a pplication rate, very mild symptoms were noted on chive, wheat, and barley; and mild symptoms on sorghum, corn and onion. The product was determined to be phytotoxic at less than or equal to half the field application rate for dicots and greater than or equal to the application rate for monocots. The study was acceptable.Killed, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 17 of 32

b. Aquatic Animals and Plants

Freshwater Fish (Harmonized Guideline 885.4200) (MRID No. 47946812). A 30-day staticrenewal laboratory study was conducted to determine the toxic effects of killed, nonviable Streptomyces acidiscabies strain RL-110 T bluegill sunfish ( Lepomis macrochirus ). The fish were exposed to nominal aqueous test material concentrations of 1.1, 11, or 28 mg dry cell mass/L of dilution water, equivalent to 1, 10, and 25 times the EEC, respectively. Additionally, the fish were exposed to a nominal dietary concentration of 110 mg/Kg (100 times the EEC). The test included a sterile filtrate control and a negative control. There was no mortality in any group, and no signs of toxicity (lesions, necroses, tumors) were observed internally or externally in any fish. The 30-day LC 50 (concentration at which 50% of the test animals are killed) and NOEL was greater than 28 mg/L (equivalent to 25 times the estimated concentration of killed, nonviable Streptomyces acidiscabies strain RL-110 T in the environment following application according to the product label). The study was acceptable.

Freshwater Invertebrates (Harmonized Guideline 885.4240) (MRID Nos. 47946813, 48104504). A 21-day static-renewal laboratory study was conducted to determine the effects of killed,

nonviable Streptomyces acidiscabies strain RL-110 T on survival and reproduction of the

freshwater invertebrate Daphnia magna . Neonates were exposed to nominal concentrations of killed, nonviable Streptomyces acidiscabies strain RL-110 T at 1.1, 11, or 28 mg dry cell mass/L of dilution water (equivalent to 1, 10, and 25 times the EEC, respectively). The test included a sterile filtrate control and a negative control of dilution water only. Survival in the 1.1, 11, and 28 mg/L groups was 95%, 65%, and 80%, respectively, compared to 90% in the negative control group. Survival in the sterile filtrate group was 55% , possibly due to bacterial contamination of the fermentation broth. Daphnids in the 11 and 28 mg dry cell mass/L groups were observed to be small and pale in color compared to the negative control group. No adverse effects were seen in the 1.1 mg/L group. The mean cumulative number of offspring per female in the 1.1, 11, and 28 mg/L groups was 151, 80, and 28, respectively, compared to 130 in the negative control group and 118 in the sterile filtrate group. The 21-day EC 50 for survival (the concentration, or “effective concentration,” of the test substance at which the survival of the test group is 50%) is greater than 28 mg dry cell mass/L of dilution water (25 times the EEC) and NOEL was greater than 1.1 mg dry cell mass/L (1 times the EEC). The study was acceptable.

In a 21-day static-renewal laboratory study conducted to determine the effect of the end-use product MBI-005 EP containing 17% killed, nonviable Streptomyces acidiscabies strain RL110 T in spent fermentation media on survival and growth of the freshwater invertebrate, Hyalella

azteca, third instars were exposed to nominal test material concentrations of 11.04, 110.4, or 1,104.0 mg/L of water, chosen to represent 10, 100, and 1000 times the expected EEC, respectively. The test included a negative control of dilution water only and a sterile filtrate control at 1,104 mg/L. Mean survival in the 11.04, 110.4, and 1,104 mg/L groups was 97.5%, 95%, and 97.5%, respectively, compared to 100% in the negative control. Mean survival in the sterile filtrate group was 92.5%, which was significantly lower than the negative control group.

The mean survival rates of the control and test material groups were not significantly different.

The EC 50 for survival was greater than 1,104 mg/L. The mean weight of the surviving organisms in the 11.04, 110.4, and 1,104 mg/L groups was 0.133, 0.129, and 0.065 mg, respectively,Killed, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 18 of 32 compared to 0.130 mg in the negative control group. Mean weight of survivors in the sterile filtrate group was 0.080 mg. The NOEL for growth was 1,057.3 mg/L. The EC 50 for survival was greater than1,104 mg/L, or 1000 times the EEC; The NOEL for growth was less than or equal to 1,057.3 mg/L. The study was acceptable.

Estuarine/Marine Fish and Invertebrates (Harmonized Guideline 885.4280), (MRID No. 47946817): The applicant provided the following rationale to support a waiver of Estuarine/Marine Fish and Invertebrates Toxicity/Pathogenicity testing: (1) killed, nonviable Streptomyces acidiscabies strain RL-110 T is not intended for direct application to estuarine or marine environments, nor is it expected to enter those environments in significant concentrations; (2) the toxicity and pathogenicity of Streptomyces acidiscabies strain RL-110 T

have been evaluated in freshwater fish and invertebrates, and the NOEL was greater than 25 times the estimated environmental concentration for fish and aquatic invertebrates; (3) Streptomyces acidiscabies strain RL-110 T is a naturally-occurring bacterium commonly found in soils; and (4) a literature search yielded no reports of adverse effects in aquatic species.

EPA considers the toxicity component of the estuarine/marine fish and invertebrates data requirement to be fulfilled by the freshwater fish toxicity/pathogenicity study (MRID 47946812) and the two studies with freshwater invertebrates (MRID Nos. 47946813 and 48104504). These

studies showed no toxic effects to fish or survival or reproductive effects on freshwater

invertebrates. Streptomyces acidiscabies strain RL-110 T is killed and nonviable, and pathogenicity and/or infectivity in estuarine and marine organisms are not expected. It would be inappropriate and unnecessary to conduct testing with this active ingredient to evaluate these endpoints. Thus, EPA waived testing for the freshwater fish toxicity/pathogenicity and invertebrate testing requirements.

Nontarget Plants (Harmonized Guideline 885.4300); (MRID No.47990702) : Bioassays were conducted to determine the effects of the end-use product MOI-005 EP containing 17% killed,

nonviable Streptomyces acidiscabies strain RL-110 T in spent fermentation media on nontarget aquatic plants. In the aquatic plant tests, MBI-005 EP was incorporated into the growth medium in a series of dilution concentrations ranging from the field application rate to 85 times lower than the field application rate for duckweed ( Lemna minor ), and from the field application rate to 700 times lower than the application rate for the green alga, Pseudokirchneriella subcapitata

. After seven days of incubation, even the most diluted test material treatments were toxic to

duckweed and P. subcapitata . The product is determined to be phytotoxic at less than or equal to one-half the application rate for aquatic plants. The study was acceptable.

2. Environmental Fate Data

The need for environmental fate and groundwater data was not triggered because results of the acute toxicity studies did not trigger any additional Tier I studies.Killed, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 19 of 32 3. Ecological Exposure and Risk Characterization

As noted in Unit IV.C.1., Streptomyces acidiscabies strain RL-110 T does not pose a hazard to nontarget animals as it is highly selective to plants.

For nontarget plants, the risk characterization is focused on thaxtomin A. The degradation potential of thaxtomin A was analyzed in one study that showed common soil fungi and bacteria readily degrade thaxtomin A and can metabolize this substance as a sole carbon source for growth. “Three bacterial isolates and two funga l isolates were grown in media containing

thaxtomin A. Growth of these organisms resulted in decreases in the optical densities at 400 nm of culture supernatants and in 10% reductions in the thaxtomin A concentration. The fungal

isolates were identified as a Penicillium sp. isolate and a Trichoderma sp. isolate. One bacterial isolate was associated with the species Ralstonia pickettii , and the two other bacterial isolates

were identified as Streptomyces sp. strains” (Doumbou et al. 1998).

In addition to the degradation of thaxtomin A, instructions on the end use product label for MBI05 provide specific directions to apply the product as a pre-, at-, or post-emergent application along with instructions on application, timing and incorporation and a precautionary statement not to apply to surface water.

Streptomyces acidiscabies strain RL-110 T does not pose a hazard to nontarget organisms when applied according to label directions, as it is highly selective to plants, requires direct application to target plants for efficacy, and is expected to degrade in the environment following application. Pesticide products containing killed, nonviable Streptomyces acidiscabies strain RL110 T are not likely to pose a risk to the environment or to any non-target organisms when applied according to the proposed label directions.

4. Endangered Species Assessment

Streptomyces acidiscabies strain RL-110 T is highly selective to plants, and requires pre- atand/or post-plant application according to label directions for efficacy. Further, it is expected to degrade in the environment when applied in accordance with the proposed product label. EPA

has determined Streptomyces acidiscabies strain RL-110 T will have "No Effect" on any currently listed threatened or endangered species or on any designated critical habitat.

V. ENVIRONMENTAL JUSTICE

EPA seeks to achieve environmental justice—the fair treatment and meaningful involvement of all people regardless of race, color, national origin, or income—with respect to the development, implementation, and enforcement of environmental laws, regulations, and policies. Fair treatment means that no group of people, incl uding racial, ethnic, or socioeconomic groups, should bear a disproportionate share of the negative environmental consequences resulting from industrial, municipal, and commercial operations or the execution of federal, state, local, and tribal environmental programs and policies. Meaningful involvement means that (1) potentially affected community residents have an appropriate opportunity to participate in decisions about a proposed activity that will affect their environment and/or health; (2) the public’s contributionKilled, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 20 of 32 can influence the regulatory agency’s decision; (3) the concerns of all participants involved will be considered in the decision-making process; and (4) the decision-makers seek out and facilitate the involvement of those potentially affected. EPA has this goal for all communities and persons

across the U.S.

To help address potential environmental justice issues, EPA seeks information on any groups or segments of the population who, as a result their location, cultural practices, or other factors, may have atypical, unusually high exposure to killed, nonviable Streptomyces acidiscabies strain

RL-110 T , compared to the general population. Please comment if you are aware of any subpopulations that may have atypical, unusually high exposure compared to the general population.

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

.

VI. RISK MANAGEMENT DECISION

Section 3(c)(5) of FIFRA provides 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 materials required to be submitted comply with the requirements of FIFRA; (3) it will perform its intended func tion without unreasonable adverse effects on the environment; and (4) when used in accordance with widespread and commonly recognized practice, it will not generally cause unreasonable adverse effects on the environment.

To satisfy criterion 1, the compositions of the killed, nonviable Streptomyces acidiscabies strain

RL-110 T pesticide products have well-known properties. EPA has no knowledge that would contradict the claims made on the MBI-005 TGAI and MBI-005 EP labels, and such pesticide products are not expected to cause unreasonable adverse effects on the environment when used according to their respective label instructions. Criterion 2 is satisfied by the current product labels, as well as the data and information presented in this document. It is believed that the killed, nonviable Streptomyces acidiscabies strain RL-110 T pesticide products will not cause any unreasonable adverse effects on the environment, and MBI-005 EP (end-use pesticide product), in particular, is likely to control labeled annual grass, broadleaf and sedge weeds as claimed, satisfying criterion 3. Criterion 4 is satisfied in that the killed, nonviable Streptomyces

acidiscabies strain RL-110 T pesticide products are not expected to cause unreasonable adverse effects when used according to label instructions. Therefore, MBI-005 TGAI and MBI-005 EP, containing killed, nonviable Streptomyces acidiscabies strain RL-110 T as a new active ingredient, are eligible for registration under FIFRA section 3(c)(5) for the labeled uses.Killed, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 21 of 32 VII. ACTIONS REQUIRED OF THE REGISTRANT

A. Final Printed Labeling

Before releasing pesticide products containing killed, nonviable Streptomyces acidiscabies strain

RL-110 T for shipment, the registrant is required to provide appropriate final printed labeling to

EPA.

B. Terms of Registration

As terms of the MBI-005 TGAI and MBI-005 EP registrations, MBI must submit the results of storage stability (Harmonized Guideline 830.6317) and corrosion characteristics (Harmonized Guideline 830.6320) studies within one year of registration and verification of OMRI

certification.

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 listed

below.

Reports of all incidents of adverse effects to the environment must be submitted to EPA under the provisions stated in FIFRA section 6(a)(2). Additionally, all incidents of hypersensitivity (including both suspected and confirmed incidents) must be reported to EPA under the provisions of 40 CFR § 158.2140(d).

VIII. BIBLIOGRAPHY

For definitions of scientific terms, please refer to http://www.epa.gov/pesticides/glossary/

.

A. Studies Submitted to Support the Registration of Pesticide Products Containing Killed,

Nonviable Streptomyces acidiscabies strain RL-110 T

MRID No. Citation

47946801 Koivunen, M. 2009. Product Chemistry for MBI-005 TGAI. Unpublished study prepared by MBI. 280 p.

47946802 Kuhn, J. 2009. MOI 005: Acute Oral Toxicity St udy (UDP) in Rats: Final Report. Project Number: 12679/09. Unpublished study prepared by Stillmeadow, Inc. 11 p.

47946803 Kuhn, J. 2009. MOI 005: Acute Dermal Toxicity Study in Rats: Final Report. Project Number: 13042/09. Unpublished study prepared by Stillmeadow, Inc. 11 p.Killed, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 22 of 32

MRID No. Citation

47946804 Younger, C. 2009. MOI 005: Acute Inhalation Toxi city Study in Rats: Final Report. Project Number: 13043/09. Unpublished study prepared by Stillmeadow, Inc. 17 p.

47946805 Kuhn, J. 2009. MOI 005: Acute Eye Irritation Study in Rabbits: Final Report. Project Number: 13044/09. Unpublished study prepared by Stillmeadow, Inc. 16 p.

47946806 Kuhn, J. 2009. MOI 005: Acute Dermal Irritation Study in Rabbits: Final Report. Project Number: 13045/09. Unpublished study prepared by Stillmeadow, Inc. 11 p.

47946807 Kuhn, J. 2009. MOI 005: Skin Sensitization Study in Guinea Pigs: Final Report. Project Number: 13046/09. Unpublished study prepared by Stillmeadow, Inc. 17 p.

47946808 Monds, K. 2009. MOI 005 Streptomyces acidiscabies : Acute Injection Toxicity/Infectivity Study (in Rats): Final Report. Project Number: 13054/09. Unpublished study prepared by Stillmeadow, Inc. 19 p.

47946809 Monds, K. 2009. MOI 005: Acute Injection Toxicity/Pathogenicity Pilot Study in Mice: Final Report. Project Number: 13095/09. Unpublished study prepared by Stillmeadow, Inc. 8 p.

47946810 Monds, K. 2009. MOI 005: Subcutaneous Mous e Safety Study: Final Report. Project Number: 13096/09. Unpublished study prepared by Stillmeadow, Inc. 8 p.

47946811 Hubbard, P.; Beavers, J. 2009. MBI-005: An Av ian Oral Pathogenicity and Toxicity Study in the Mallard. Project Number: 664/102. Unpublished study prepared by Wildlife International, Ltd. 50 p.

47946812 Sayers, L. 2009. MBI 005: 30 Day Bluegill Sunfish ( Lepomis macrochirus ) Static-Renewal Toxicity, Infectivity and Pathogenicity Test. Project Number: 071409/OPPTS885/BG/STR, 13909/6103. Unpublished study prepared by Springborn Smithers Laboratories. 46 p.

47946813 Sayers, L. 2009. MBI 005: 21 Day Daphnid ( Daphnia magna ) Static Renewal Toxicity, Infectivity and Pathogenicity Test. Project Number: 13909/6104, 071409/OPPTS/DAPHNID/STR/MPCA. Unpublis hed study prepared by Springborn Smithers Laboratories. 45 p.

47946814 Porch, J.; Krueger, H. 2009. MBI 005: A Dietary Pathogenicity and Toxicity Study with the Ladybird Beetle ( Hippodamia convergens ): Final Report. Project Number: 664/104, 664/072109/LBBLCP/SUB664. Unpublished study prepared by Wildlife International, Ltd. 30 p.

47946815 Porch, J.; Krueger, H. 2009. MBI 005: A Dietar y Pathogenicity and Toxicity Study with Green Lacewing Larvae ( Chrysoperla rufilabras ): Final Report. Project Number: 664/105, 664/072109/GLWLCP/SUB664. Unpublished study prepared by Wildlife International, Ltd. 45 p.Killed, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 23 of 32

MRID No. Citation

47946816 Richards, K. 2009. Evaluation of the Dietary Effect(s) of MBI 005 on Adult Honey Bees ( Apis mellifera L.). Project Number: CAR/139/09. Unpublished study prepared by California Agricultural Research, Inc. 68 p.

47946817 Pitts, K. 2009. Response to Tier 1 Microbial Pesticide Data Requirements for MBI-005 TGAI. Unpublished study prepared by MBI. 29 p.

47946818 Pitts, K. 2009. Endangered Species Evaluation for MBI-005. Unpublished study prepared by MBI. 80 p.

47946819 Cordova-Kreylos, A. 2009. MBI-005: Literature Review: Scab-Causing Streptomyces

. Unpublished study prepared by MBI. 294 p.

47990702 Morgan, C.; Koivunen, M. 2010. MBI-005 - Nontarget Plants. Project Number: 1503/33. Unpublished study prepared by MBI, Inc. 63 p.

48104501 Koivunen, M. 2010. Secondary Metabolites of MBI-005 ( Streptomyces acidiscabies ). Unpublished study prepared by MBI. 128 p.

48104502 Pitts, K. 2010. One Month Interim Report for Storage Stability and Corrosion Characteristics of MBI-005 TGAI. Unpublished study prepared by MBI. 46 p.

48104503 Koivunen, M. 2010. Manufacturing Process fo r MBI-005 TGAI: Heat Devitalization Process: Supplemental to MRID 47946801. Unpublished study prepared by MBI. 15 p.

48104504 Hartwell, T. 2010. MOI 005: Microbial Pest C ontrol Agent (MCPA) Freshwater Aquatic

Invertebrate Test with Hyalella azteca : Final Report. Project Number: 13841/10. Unpublished study prepared by Stillmeadow, Inc. 17 p.

48104505 Younger, C. 2010. MBI 005: Parasitic Wasp Non- Target Insect Microbial Testing. Project Number: 13483/09. Unpublished study prepared by Stillmeadow, Inc. 14 p.

48140901 Koivunen, M. 2010. Product Chemistry for MBI-005 EP. Unpublished study prepared by MBI. 190 p.

48140902 Pitts, K. 2010. Response to Tier 1 Microbial Pesticide Data Requirements for MBI-005 EP. Unpublished study prepared by MBI. 88 p.

48582301 Tan, E. 2011. Amended Manufacturing Process for MBI-005 TGAI. Unpublished study prepared by MBI. 52p.

48582302 Tan, E. 2011. Heat Inactivation of MBI-005 TGAI (Streptomyces acidiscabies strain RL110). Unpublished study prepared by MBI. 25p.

48582401 Tan, E. 2011. Amended Manufacturing Process for MBI-005 EP. Unpublished study prepared by MBI. 59p.Killed, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 24 of 32

B. Environmental Protection Agency Risk Assessment Memoranda

U.S. EPA. 2011a. Streptomyces acidiscabies strain RL-110 T (ATCC 49003) producing thaxtomin A. MOI-401. Memorandum from J.V. Gagliardi, Ph.D. and J.L. Kough, Ph.D. to A. Sibold dated Nov. 15, 2011.

U.S. EPA. 2011b. Streptomyces acidiscabies strain RL-110 T (ATCC 49003) producing thaxtomin A. MOI-401. Memorandum from J.V. Gagliardi, Ph.D. and J.L. Kough, Ph.D. to A. Sibold dated Jan. 25, 2012.

C. Other References

Bischoff, V., S.J. Cookson, S. Wu and W.R. Scheible. 2009. Thaxtomin A affects CESA- complex density, expression of cell wall genes, cell wall composition, and causes ectopic lignification in Arabidopsis thaliana seedlings. Journal of Experimental Botany 60(3):955-

65.

Doumbou, C.L., V. Akimov and C. Beaulieu. 1998. Selection and characterization of microorganisms utilizing thaxtomin A, a phytotoxin produced by Streptomyces scabies

. Applied and Environmental Microbiology 64(11):4313-4316.

Faucher, E., T. Savard, and C. Beaulieu. 1992. Char acterization of Actinomycetes Isolated from Common Scab Lesions on Potato-Tubers. Canadian Journal of Plant Pathology-Revue Canadienne De Phytopathologie. 14: 197-202.

Fett W.F., et al . 1992. Cutinase production by Streptomyces spp.. Curr

. Microbiol . 25:165-171.

Lambert, D.H., and R. Loria. 1989. Streptomy ces-Acidiscabies SP-NOV. International Journal of Systematic Bacteriology 39:393-396.

Loria, R., D.R. D. Bignell, S. Moll, J.C. Huguet-Tapia, M.V. Joshi, E.G. Johnson, R.F. Seipke, and D.M. Gibson. 2008. Thaxtomin biosynthesis: The Path to Plant Pathogenicity in the Genus Streptomyces. Antonie Van Leeuwenhoek International Journal of General and Molecular Microbiology 94:3-10.

Loria, R., J. Kers, and M. Joshi. 2006. Evolution of Plant Pathogenicity in Streptomyces. Annual Review of Phytopathology 44:469-487.

Murray, P.R., E.J. Baron, J.H. Jorgensen, M.L. Landry and M.A. Pfaller, editors. 2007. Manual of Clinical Biology. Vol.1. 9 th Ed. Washington (DC): ASM Press.

Wanner, L.A. 2009. A Patchwork of Streptomyces Species Isolated from Potato Common Scab

Lesions in North America. American Journal of Potato Research 86:247-264.Killed, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 25 of 32

APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS (40 CFR PART 158 – SUBPART V)

TABLE 1. Product Analysis Data Requirements for the Technica l Grade of the Active Ingredient (TGAI), Killed, Nonviable Streptomyces acidiscabies strain RL-110 T , and Its Associated End-Use Pesticide Product (EP), MBI-005 EP (40 CFR § 158.2120)

Harmonized

Guideline

Number Data Requirement Results MRID No. TGAI EP

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

MBI-005 TGAI contains 100% by weight killed, nonviable Streptomyces acidiscabies strain

RL-110 T cells and spent

fermentation media. Submitted data fulfill the requirement for product identity. MBI-005 EP contains 17% killed, nonviable Streptomyces acidiscabies

strain RL-110 T cells and spent

fermentation media.

47946801

48140901

48104503

48582301

48582302

48582401 885.1200 Manufacturing Process Submitted data fulfill th e requirement for manufacturing process.

885.1300 Discussion of Formation

of Unintentional Ingredients Submitted data fulfill the requirement for discussion of formation of unintentional ingredients.

N/A Deposition of a Sample in a Nationally Recognized Culture

Collection A sample of Streptomyces

acidiscabies strain RL-110 T is on deposit with the American Type Culture Collection under

Accession Number ATCC

49003. N/A

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

885.1500 Certification of Limits Limits are adequate/acceptable.Killed, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 26 of 32

TABLE 2. Physical and Chemical Charact eristics for the Technical Grade of the Ac tive Ingredient (TGAI), Killed, Nonviable Streptomyces acidiscabies strain RL-110 T , and Its Associated End-Use Pesticide Product (EP), MBI-005 EP (40 CFR § 158.2120)

Harmonized

Guideline Number Data Requirement Results MRID No. TGAI EP

830.6302 1 Color Amber Black

47946801

48104502

48140901 830.6303 1 Physical State Liquid

830.6304 1 Odor Grass-like Rancid, Sweet

830.6313 1 Stability to Normal and

Elevated Temperatures, Metals,

and Metal Ions Expected to be stable when

stored at 25°C. N/A

830.6317 Storage Stability Expected to be stable for one year when stored per label

directions. As a term of the MBI-005 TGAI registration, EPA is requiring submission of a storage stability study within one year of registration. Expected to be stable for two years when stored per label

directions. As a term of the MBI-005 EP registration, EPA is requiring submission of a storage stability study within one year of registration.

830.6319 Miscibility Not required because MBI-005

TGAI is not an emulsifiable liquid form of a microbial pesticide (refer to test note #2 of 40 CFR § 158.2120(d)). Miscible with water.

830.6320 Corrosion

Characteristics Not expected to be corrosive to packaging materials. As terms of the MBI-005 TGAI and MBI-005 EP registrations, EPA is requiring

submission of corrosion characteri stics studies within one year of registration.

830.7000 1 pH 7.4 6.6 830.7100 Viscosity 4.96 cP 63.8 cP

830.7300 1 Density/Relative Density/Bulk Density

(Specific Gravity) 0.999 g/mL 1.02 g/mL

1 According to 40 CFR § 158.2120, these data are only required for th e technical grade of the activ e ingredient. Since Marrone Bi o Innovations included this information with its applicati on for MBI-005 EP, it is summarized appropriately in this table.Killed, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 27 of 32

TABLE 3. Toxicology Data Requirements for the Technical Gr ade of the Active Ingredient (TGAI), Killed, Nonviable S treptomyces
acidiscabies strain RL-110 T , and Its Associated End-Use Pro duct (EP), MBI-005 EP (40 CFR § 158.2140)

Harmonized

Guideline Number Data Requirement Results MRID No. TGAI EP

Tier I

885.3050 Acute Oral Toxicity/ Pathogenicity Toxicity component of the acute oral toxicity/pathogenicity data requirement fulfilled by MRID No.

479468-02. Given that Streptomyces

acidiscabies strain RL-110 T is killed and nonviable, pathogenicity and/or infectivity in mammals is not expected. It would be inappropriate and unnecessary to conduct testing with this active ingredient to evaluate these endpoints. Thus, EPA waived testing for the pathogenicity component of the acute oral toxicity/pathogenicity data requirement. Classification: Acceptable N/A 47946817

885.3150 Acute Pulmonary Toxicity/ Pathogenicity Toxicity component of the acute pulmonary toxicity/pathogenicity data requirement fulfilled by MRID No.

479468-04. Given that Streptomyces

acidiscabies strain RL-110 T is killed and nonviable, pathogenicity and/or infectivity in mammals is not expected. It would be inappropriate and unnecessary to conduct testing with this active ingredient to evaluate these endpoints. Thus, EPA waived testing for the pathogenicity component of the acute pulmonary toxicity/pathogenicity data requirement. Classification: Acceptable N/A 47946817

885.3200 Acute Injection Toxicity/ Pathogenicity (Intravenous) Not toxic, infective, or pathogenic (live Streptomyces acidiscabies strain RL-110 T ) when administered intravenously at 9.0 x

10 6 CFU per animal. Classification: Acceptable N/A 47946808

885.3400 Hypersensitivity

Incidents Hypersensitivity incidents, including imme diate-type and delayed-type reactions of humans or domestic animals, that occu r during the testing or production of the TGAI, MP or EP, or are otherwise known to the applicant, must be reported if they occur (refer to test note #3 of 40 CFR § 158.2140(d)). N/A

885.3500 Cell Culture Not required because killed, nonviable Streptomyces acidiscabies strain RL-110 T
is not a virus (refer to test note #4 of 40 CFR § 158.2140(d)). N/A N/AKilled, nonviable Streptomyces acidiscabies strain RL-110 T

Biopesticides Registra tion Action Document Page 28 of 32

TABLE 3. Toxicology Data Requirements for the Technical Gr ade of the Active Ingredient (TGAI), Killed, Nonviable S treptomyces
acidiscabies strain RL-110 T , and Its Associated End-Use Pro duct (EP), MBI-005 EP (40 CFR § 158.2140)

Harmonized

Guideline Number Data Requirement Results MRID No. TGAI EP

870.1100 Acute Oral Toxicity Oral LD 50 female rats > 5,000 mg/kg Classification: Acceptable

TOXICITY CATEGORY IV Waived based on the results of

MRID No. 479468-02 and because

MBI-005 EP contains inert ingredients that are not expected to be of toxicological concern (refer to test note #5 of 40 CFR § 158.2140(d)). Classification: Acceptable Toxicity Category III 47946802

48140902

870.1200 Acute Dermal Toxicity Dermal LD 50 combined (male and female) rats > 5,050 mg/kg Classification: Acceptable

TOXICITY CATEGORY IV Waived based on the results of

MRID No. 479468-03 and because

MBI-005 EP contains inert ingredients that are not expected to be of toxicological concern (refer to test note #5 of 40 CFR § 158.2140(d)). Classification: Acceptable Toxicity Category III 47946803

48140902

870.1300 Acute Inhalation Toxicity Inhalation LC 50 combined (male and femal e rats > 2.12 mg/L Classification: Acceptable

TOXICITY CATEGORY IV Waived based on the results of

MRID No. 479468-04 and because

MBI-005 EP contains inert ingredients that are not expected to be of toxicological concern (refer to test note #5 of 40 CFR § 158.2140(d)). Classification: Acceptable Toxicity Category III 47946804

48140902

870.2400 Acute Eye

Irritation MOI 005 was minimally irritating to the eyes of rabbits. Classification: Acceptable

TOXICITY CATEGORY IV Waived based on the results of

MRID No. 479468-05 and because

MBI-005 EP contains inert ingredients that are not expected to be of toxicological concern (refer to test note #5 of 40 CFR § 158.2140(d)). Classification: Acceptable Toxicity Category III 47946805

48140902

870.2500 Primary Dermal

Irritation MOI 005 was not irritating to the skin of

rabbits. Classification: Acceptable

TOXICITY CATEGORY IV Waived based on the results of

MRID No. 479468-06 and because

MBI-005 EP contains inert ingredients that are not expected to be of toxicological concern (refer to test note #5 of 40 CFR § 158.2140(d)). Classification: Acceptable Toxicity Category III 47946806

48140902

870.2600 Skin Sensitization Not a dermal sensitizer Classification: Acceptable N/A 47946807

Tiers II and III Not required for killed, nonviable Streptomyces acidiscabies strain RL-110 T , based on the lack of acute toxicity in the Tier I studies.Table 4. Non-target Organism Toxicity Requirements for Streptomyces acidiscabies strain RL-110T(40 CFR 158.2060Requirement(OPPTSGuideline No.)MRID Toxicity and ClassificationAvian oraltoxicity(885.4050)47946811 An indoor study was conducted to determine the acute toxicity of killed, nonviableStreptomyces acidiscabiesstrain RL-110Tto the mallard(Anas platyrhynchos).Fourteen-day-old birds were dosed with the test material (63 mg a.i./Kg body weight) once daily for five days andobserved for an additional 25 days. There were no treatment related signs of toxicity in any of the birds during the study. TheNOEL is greaterthan 63 mg a.i./Kg body weight/day for five days.Classification: Acceptable.Avian inhalationtoxicity/pathogenicity(885.4100)47946817“Data required when the nature of the microbial pesticide and/orits toxins indicates potential pathogenicity to birds”. (40 CFR 158.2150,testnote 3).MBI submitted a request for a waiver of the conditional data requirement based on the rationale that (1) killed, nonviableStreptomycesacidiscabiesstrain RL-110Twas not toxic or pathogenic in an avian oral toxicity study (2) the active ingredient in MBI-005 TGAI isubiquitous as a naturally occurring bacterium found in soils and water and 3) A literature search yielded no reports of adverseeffects in avianspecies.Classification: The waiver rationale is acceptable.Wild mammaltoxicity/pathogenicity(885.4150)47946817This study is “Required on a case-by-case basis if results oftests required by §158.2140 are inadequate or inappropriate for assessment ofhazards to wild mammals” (40 CFR 158.2150, testnote 4).MBI submitted a request for a waiver of the conditional data requirement based on the rationale that (1) killed, nonviableStreptomycesacidiscabiesstrain RL-110Twas not toxic in acute exposure studies in rats, (2)Streptomyces acidiscabiesstrain RL-110Twas not pathogenicin an intravenous injection study in rats, (3) the active ingredient in MBI-005 TGAI is ubiquitous as a naturally occurring bacterium found insoils; and 4) a literature search yielded no reportsof adverse effects in humans or other mammals.Classification: The waiver rationale is acceptable. Effects on wildmammals are expected to be minimal as extensive animal testing occurredon both the viable and non-viable active ingredient with no effects.Freshwater fishtoxicity/pathogenicity(885.4200)47946812 A 30-day static-renewal laboratory study was conducted todetermine the toxic effects of killed, nonviableStreptomyces acidiscabiesstrainRL-110Tto bluegill sunfish (Lepomis macrochirus). There was no mortality in any group, and no signs of toxicity (lesions, necroses, tumors)were observed internally or externally in any fish. 30-day LC50and NOEL was greater than28 mg/L (25 times greater than the estimatedenvironmental concentration (EEC)).Classification: AcceptableFreshwaterinvertebratetesting(885.4240)47946813 A 21-day static-renewal laboratory study was conductedto determine the effects of killed, nonviableStreptomyces acidiscabiesstrain RL110Ton survival and reproduction of Daphnia magna. The 21-day EC50 isgreater than 28 mg dry cell mass/L (25X the EEC) and NOEL isgreater than 1.1 mg dry cell mass/L (equivalent to the EEC).Classification: Acceptable48104504. A 21-day static-renewal laboratory study was conducted to determine the effect of the end use product MBI-005 EP onsurvival and growth ofthe freshwater invertebrate,Hyalella azteca. The NOEL for growth was 1,057.3 mg/L. The EC50 for survival was greater than1,104 mg/L.NOEL for growth was less than or equal to 1,057.3 mg/L.Classification: AcceptableEstuarine/Marinefish testingEstuarine/Marineinvertebratetesting(885.4280)47946817This study is “Required if the product is intended for direct application into the estuarine or marine environment or expectedto enter thisenvironment in significant concentrations because of expected use or mobility pattern.” (40 CFR 158.2150, testnote 6).MBI submitted a request for a waiver of the conditional data requirement based on the rationale that 1) The end use product MBI-005 EP isnot intended for direct application to estuarineor marine environments; 2) killed, nonviableStreptomyces acidiscabiesstrain RL-110Ttoxicityand pathogenicity have been evaluated in freshwater fish and invertebrates, 3) the active ingredient is ubiquitous as a naturally-occurringbacterium found in soils, and 4) a literature search yielded no reports of adverse effects in aquatic species.Classification: The waiver rationale is acceptable. Effects to estuarine and marine organisms are expected to be minimal aquatic organismtesting occurred on the non-viable TGAI and EP products.Nontarget PlantTesting(885.4300)47990702 Bioassays were conducted to determine the effects of the end use product MBI-005 EP on nontarget crop and aquatic plants. MBI-005 EP wasphytotoxic to all dicots tested, even at the 0.5X rate. Phytotoxicity was mild to moderate on broccoli, tomato, and pea; moderate on lettuce andbean; and moderate to severe on soybean and cucumber. For monocots no phytotoxicity was seen on bluegrass, bentgrass, or rice.At thehighest application rate, very mild symptoms were seen on chive, wheat, and barley; and mild symptoms on sorghum, corn and onion. In theaquatic plant tests, MBI-005 EP was incorporated in the growthmedium in a series of dilution concentrations ranging from the fieldapplication rate to 85 times lower than the field application rate for duckweed (Lemna minor) and from the field application rate to 700 timeslower than the application rate for the green algaPseudokirchneriella subcapitata. After seven days of incubation, even the most diluted testmaterial treatments were toxic to duckweed andP. subcapitata.Classification: Acceptable – The product is phytotoxic at less thanor equal to half the field application rate for dicots; greater than or equal tothe application rate for monocots; less than or equalto one half the application rate for aquatic plants.Killed, nonviableStreptomyces acidiscabiesstrain RL-110TBiopesticides Registration Action DocumentPage 31 of 32Nontarget InsectTesting(885.4340)47946814 A laboratory study was conducted to determine the effects ofkilled, nonviableStreptomyces acidiscabiesstrain RL-110Tto adult ladybirdbeetles (Hippodamia convergens). Surviving beetles in all groups were normalin appearance and behavior throughout the test.Classification: Acceptable – The NOEL for 17 day diet exposure was 51,000 ppm.47946815 A laboratory study was conducted to determine the toxic effects ofkilled, nonviableStreptomyces acidiscabiesstrain RL-110Tto greenlacewing (Chrysoperla rufilabris) larvae. There were no signs of toxicity.Classification: Acceptable – The NOEL for 10 day diet exposure was 51,000 ppm.48104505 A laboratory study was conducted to determine the toxic effects of the end use product containing 17%killed, nonviableStreptomycesacidiscabiesstrain RL-110Tand spent fermentation media on the parasitic wasp. The testmaterial did not appear to be toxic to the parasiticwasp when applied to plant hosts at 10X the field application rate.Classification: Acceptable. TheNOEL was greater than 11.04 mg/L.HoneybeeTesting(885.4380)47946816 A laboratory study was conducted to determine the effects of MBI-005 TGAI in the diet to adult honeybees (Apis mellifera). There were noadverse effects of MBI-005 on adult honeybees under the conditions of this study.Classification: Acceptable – The NOEL was 500 μg/mL in 30% sucrose solution for 20 days.APPENDIX B. PESTICIDE PRODUCTSEPA File SymbolRegistration NamePercentage ActiveIngredientFormulation TypeUse Site(s)Method(s) ofApplicationApplication RateTarget Pest(s)84059-RR MBI-005 TGAI 100.00% Technical N/A N/A N/A N/A84059-RE MBI-005 EP 17% End Use – Liquid Listed agriculturalcrops, turf andornamentals inlandscapes,residential lawnsAerial and GroundApplication,Broadcast,Pre- plant surfaceapplication,incorporated,Pre-plantincorporated,At-plant,incorporated, andLayby, soil directed.Up to 12 quarts of EPper 1000 square feetLabeled annualgrasses, broadleaf andsedge weeds.