BIOPESTICIDES REGISTRATION ACTION DOCUMENT



## Bacillus thuringiensis subsp. kurstaki strain VBTS-2546

Pesticide Chemical (PC) Code: 006699

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

DRAFT

September 4, 2012Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 2 of 38

TABLE OF CONTENTS

I. EXECUT IVE SU MMARY

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

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

III. REGULATORY BACKGROUND .................................................................................................. ............ 5

A. Applications for Pe sticide Registration ................................................................................... ....... 5

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

IV. RISK ASSESSMENT ............................................................................................................. ..................... 6

A. Product Analysis Assess ment (40 CFR § 158.2120) ....................................................................... 6

B. Human Health Risk Asse ssment (40 CFR 158.2140) ..................................................................... 8

C. Environmental Assessmen t (40 CFR § 15 8.2150) ........................................................................ 15

V. ENVIRONMENTAL JUSTICE ......................................................................................................... ........ 19

VI. RISK MANAGEMENT DECISION ...................................................................................................... ... 19

VII. ACTIONS REQUIRED OF THE REGISTRANT ................................................................................... 20

A. Final Prin ted Labeling .................................................................................................... ............... 20

B. Terms of Registra tion ..................................................................................................... ................ 20

C. Reporting of Adverse Effects and Hypersensitivi ty Incidents .................................................... 20

VIII. BIBLIOGRAPHY .................................................................................................................. ..................... 21

A. Studies Submitted to Support the Regist ration of Pesticide Products Containing Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 ....................................................................................... 21

B. Environmental Protection Agen cy Risk Assessment Memoranda ............................................. 22

C. Other Referen ces .......................................................................................................... .................. 23

APPENDIX A. MICROBIAL PESTIC IDES DATA REQUIREMENTS ........................................................... 25

APPENDIX B. PESTICIDE PRODUCTS ............................................................................................... ............... 38Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 3 of 38

BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

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

Science Review

Judy Facey, Ph.D. Product Analysis and Human Health Shannon Borges, M.S. Nontarget Effects

Regulatory Review

Ann Sibold, M.A. Regulatory Action Leader, MPB Shannon Borges, M.S. Acting Team Leader,

MPB Kimberly Nesci, M.S. Acting Chief, MPBBacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 4 of 38

I. EXECUTIVE SUMMARY

Description of the Active Ingredient: Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 is a Gram positive, spore forming, rod-shaped bacterium, that produces crystalline protein

inclusions that are toxic to larvae of some Lepidopteran insects upon inge stion. The strain was obtained from an acquired collection of domestic Bacillus thuringiensis isolates. Analysis of the microarray indicates that strain VBTS-2546 has the following genes: cry1Aa, cry1Ab, cry1Ac, cry2Aa, cry2Ab, cry1Ia, vip3Aa . These same genes are identified in the highly similar pattern reported for the currently registered Bacillus thuringiensis subsp. kurstaki strain HD-1. HD-1 is synonymous with the active ingredient, Bacillus thuringiensis subsp. kurstaki strain ABTS-351,

which is contained in Va lent’s DiPel® products.

Proposed products containing Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 are as

follows:

VBC-60219 Biological Insecticide Slurry (73049-UT I) is the fermentation slurry concentrate, proposed for manufacturing use only, containing the active ingredient Bacillus thuringiensis subsp. kurstaki strain VBTS-2546. This slurry is used in the manufacturing of the technical powder (73049-UTO) and end use products.

VBC-60225 Biological Insecticide Technical Po wder Active Ingredient (73049-UTO) is the technical powder, proposed for manufacturi ng use only, containing th e active ingredient Bacillus thuringiensis subsp. kurstaki strain VBTS-2546. This technical powder is not currently used in any end use product. It is prepared for testi ng in the mammalian health and nontarget organism studies to evaluate potential hazards. The technical powder has the highest content of fermentation solids, spores and insecticidal toxins.

Two proposed end use products containing Bacillus thuringiensis subsp. kurstaki, strain VBTS2546 as active ingredient (VBC-60236 Biological In secticide Dry Flowable and Foray 2WG) (EPA File Symbols 73049-UIN and 73049-UIR respec tively) are effective against the listed economically important Lepidopteran larvae pr esent on a large variet y of food and non-food crops and in forestry.

Assessing Risks to Human Health: Data derived from appropriate tests give no indications that the living microorganism is toxic or pathogenic to humans. No harmful health effects to humans are expected from its use as a pesticide.

Assessing Risks to the Environment: Pesticide products containing Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 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 decision 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.Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 5 of 38

Consistent with the policy of making registration decisions more transparent, the proposed pesticide products containing a new active ingredient, Bacillus thuringiensis subsp. kurstaki strain VBTS2546 , will be subject to a comment period. In addition to containing a new active ingredient, the registration of these pesticide products would result in the first outdoor and food uses for Bacillus thuringiensis subsp. kurstaki strain VBTS-2546

. The following documents are being made available for comment in the docket, id entification number EPA-HQ-OPP-2011-0769, and

accessed through either http://www.regulations.gov or http://www.epa.gov/pesticides/regu lating/registrati on-status.html : (1) draft Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 Bi opesticides Registration Action Document (BRAD); (2) draft label for the manufactur ing-use pesticide product, VBC-60219 Biological Insecticide Slurry (EPA File Symbol 73049- UTI); (3) draft label for VBC-60225 Biological Insecticide Technical Powder Active Ingredient; (4) draft labels for the two end-use pesticide products, VBC-60236 Biological Ins ecticide Dry Flowable (EPA File Symbol 73049-UIN) and Foray 2WG Biological Insecticide (EPA File Symbol 73049-UIR).

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

.

II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 ( Btk)

Culture Collection: Deposited at the American Type Culture Collection (ATCC) in Manassas, Virginia under ATCC SD-6283.

OPP Chemical Code: 006699, formerly 006402

Type of Pesticide: Microbial Pesticide – Insecticide

See Appendix B for specific information (i.e., use sites, application rates, methods of application, form ulation types, and target pests) regarding the proposed pesticide pr oducts containing this active ingredient.

III. REGULATORY BACKGROUND

A. Applications for Pesticide Registration

On August 1, 2011, Valent BioSciences Corpor ation, 870 Technology Way, Libertyville, IL 60048 submitted an application for registration of four products, the manuf acturing-use pesticide product, VBC-60219 Biological Insecticide Slur ry (EPA File Symbol 73049-UTI); VBC-60225 Biological Insecticide Technical Powder Active Ingredient (EPA File Symbol 73049-UTO); and two end-use pesticide products, VBC-60236 Biologi cal Insecticide Dry Flowable (EPA File Symbol 73049-UIN) and Foray 2W G Biological Insecticide (E PA File Symbol 73049-UIR), containing the new microbi al active ingredient Bacillus thuringiensis subsp. kurstaki strain

VBTS-2546. In the Federal Register (FR) of October 12, 2011 (76 FR 63299), EPA announced receipt of these applications for registration of pesticide pr oducts containing a new activeBacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 6 of 38

ingredient for use in forests and on shade, orname ntal and sugar maple trees; on ornamental fruit, nut and citrus trees intended for aesthetic purpos es or climatic modification that are grown in interior plantscapes, ornamental gardens or pa rks; on golf courses, lawns and grounds; and on agricultural crops (veg etables, fruit trees, berry bushes, nut trees, grains, herbs, alfalfa, cotton, flowers, bedding plants, ornamentals, hops, a nd greenhouse, shadehouse, outdoor nursery crops, and aquatic ornamentals). No co mments were received following pub lication of this notice. For more information, see the following docket at http://www.regulations.gov/ : EPA-HQ-OPP-2011-

0769.

B. Food Tolerance Exemption

An exemption from the requirement of a tolera nce is established for viable spores of the microorganism Bacillus thuringiensis Berliner in or on honey and honeycomb and all other raw agricultural commodities when it is applied either to growing crops, or when it is applied after harvest in accordance with good agri cultural practices, provided the sp ecifications set forth in the exemption are met ( 40 CFR 180.1011 ). The specifications are listed below:

(1) The microorganism shall be an authentic strain of Bacillus thuringiensis Berliner conforming to the morphological and biochemical characteristics of Bacillus thuringiensis as described in Bergey's Manual of Determinative Bacteriology, Eighth

Edition.

(2) Spore preparations of Bacillus thuringiensis Berliner shall be produced by pure culture fermentation procedures with adequate contro l measures during production to detect any changes from the characteristics of the parent strain or contamination by other microorganisms.

(3) Each lot of spore preparation, prior to the addition of other materials, shall be tested by subcutaneous injection of at least 1 million s pores into each of five laboratory test mice weighing 17 grams to 23 grams. Such test shal l show no evidence of infection or injury in

the test animals when observed for 7 days following injection.

(4) Spore preparations shall be free of the Bacillus thuringiensis β -exotoxin when tested with the fly larvae toxicity test (“Microbial Control of Insects and Mites,” R.P.M. Bond et al., p. 280 ff., 1971). This specification can be satis fied either by determining that each master seed lot brought into production is a Bacillus thuringiensis strain which does not produce β -exotoxin under standard manufactur ing conditions or by periodically determining that β -exotoxin synthesized during spor e production is eliminated by the subsequent spore-harvesting procedure.

Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 meets th e criteria of 40 CFR 190.1011.

IV. RISK ASSESSMENT

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 data.

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

.Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 7 of 38

Product chemistry and composition data include in formation about the identity of the active ingredient, the manufacturing proc ess, deposition of a sample of the microbial active ingredient in a nationally recognized culture collection, a nd discussion of the potential for formation of unintentional 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.

Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 (EPA File Symbol 73049- UTI; 73049- UTO; 73049-UIR and 73049-UIN) is a Gram positive, spore forming, rod-shaped bacterium, that produces crystalline protein inclus ions that are toxic to larvae of some Lepidopteran insects upon ingestion. The strain was obtained from an acquired collection of domestic Bacillus thuringiensis isolates. Analysis of the mi croarray indicates strain VBTS -2546 has the following genes: cry1Aa, cry1Ab, cry1Ac, cry2Aa, cry2Ab, cry1Ia, vip3Aa . These same genes are identified in the highly similar pattern seen for the Bacillus thuringiensis subsp. kurstaki strain ABTS-351 contained in DiPel® product HD-1.

VBC-60219 Biological Insecticide Slurry (73049-UT I) is the fermentation slurry concentrate, proposed for manufacturing use only, containing the active ingredient Bacillus thuringiensis subsp. kurstaki strain VBTS-2546. This slurry is used in the manufacturing of the technical powder (73049-UTO) and end use products.

VBC-60225 Biological Insecticide Technical Po wder Active Ingredient (73049-UTO) is the technical powder, proposed for manufacturi ng use only, containing th e active ingredient Bacillus thuringiensis subsp. kurstaki strain VBTS-2546. This technical powder is not currently used in any end use product. It is prepared for testi ng in the mammalian health and nontarget organism studies to evaluate potential hazards. The technical powder has the highest content of fermentation solids, spores, and insecticidal toxins.

Two proposed end use products containing Bacillus thuringiensis subsp. kurstaki strain VBTS2546 as active ingredient (VBC-60236 Biological In secticide Dry Flowable and Foray 2WG) (EPA File Symbols 73049-UIN and 73049-UIR respec tively) are effective against the listed economically important Lepidopteran larvae pr esent on a large variet y of food and non-food crops and in forestry.

A new, unique PC code 006699 has been assigned to Bacillus thuringiensis subsp. kurstaki strain VBTS-2546, as it is a new active ingredient. It replaces the PC code 006402, which is associated

with several Bacillus thuringiensis Berliner active ingredients.

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 requireme nts required for registration of Bacillus thuringiensis subsp.

kurstaki strain VBTS-2546 have been fulfilled.Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 8 of 38

B. Human Health Risk Assessment ( 40 CFR 158.2140 )

1. Toxicity

All toxicology data requirements for Bacillus thuringiensis subsp. kurstaki strain VBTS-2546

have been fulfilled. Acceptable Tier I mammalian toxicol ogy data and information support the registration of the proposed pestic ide products. Tier II and Tier I II 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): In lieu of testing Bacillus thuringiensis subsp. kurstaki strain VBTS-2546, th e applicant provided the following rationale to addr ess the data requirement for acu te oral toxicity/pathogenicity testing:

Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 is characte rized by comparison with the

microbial strain contained in the DiPel® product HD-1 containing Bacillus thuringiensis subsp.

kurstaki strain ABTS-351. Plasmid separation on agar ose gels was used to compare the VBTS2546 strain to known Bt subsp. kurstaki and aizawai strains, in order to determine relatedness to those serotypes. The plasmid data strongly imply that Bacillus thuringiensis subsp. kurstaki

VBTS-2546 is a Bt kurstaki strain. Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 is a Gram positive, spore forming, rod-shaped bacterium, that produces crystalline protein inclusions that are toxic to larvae of some Lepidopteran insects upon ingestion. The strain was obtained from an acquired collection of domestic Bt isolates. In a study provided by the applicant from published scientific lite rature, the method of Letowski et al. (2005) 2 was used to evaluate Btk VBTS-2546 with slight vari ations. For this study 3 , a newer, more comprehensive version of the published microarray, designated "Crickmore vers ion 2," was used. The array showed 2546 to have the following genes: crylAa

, crylAb

, cry1Ac

, cry2Aa

, cry2Ab

, cry1Ia

, vip3Aa and to have the same cry gene profile as DiPel HD-1.

Bacillus thuringiensis subsp. kurstaki , Strain VBTS-2546 was evaluated for acute oral toxicity at the limit dose of 5000 mg/kg body weight in rats. 4 The LD 50 from this study is higher than the

limit dose.

In an acute oral toxicity/p athogenicity study conducted with DiPel HD-1 Technical in rats 5 , a single dose of approximately 10 9 CFU/kg of body weight (kg/bw ) of the test microbe was administered to Sprague-Dawley rats. No anim als died during the te st period. There was no

2 Letowski et al. (2005) Assessment of cry1 gene contents of Bacillus thuringiensis strains by use of DNA microarrays. Appl. Environ. Microbiol . 71 (9): 5391-5398. 3 T. Benson, 2011, Microbial Characterization of Bacillus thuringiensis subsp. kurstaki, Strain VBTS-2546, Report No. VBTS-2546-2011-1 (submitted) 4 C. Lowe, 2011. VBC-60225: Acute Oral Toxicity Up and Down Procedure in rats, Report No. 31269 (submitted) 5 N. Carter, 1993, Acute oral toxicity and infectivity/pathologenicity to rats of ABG 6345, Report no. G-7239.222 (MRID 41443401)Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 9 of 38

apparent treatment related toxicity or evidence of pathogenicity. A linear regression analysis of

counts from feces indicated that tota l clearance would occur by 47 days.

Based on the information presented and the l ack of toxicity/pathoge nicity via pulmonary 6 and

intravenous 7 administration, Valent BioSciences Corpor ation is requesting th at EPA agree that no further testing via oral administration of Bacillus thuringiensis subsp. kurstaki strain VBTS2546 is necessary to support registration. Intravenous administ ration and pulmonary

administration are considered tests that are more likely to present toxicity or pathogenicity. Valent cited the oral administration study for Bacillus thuringiensis subsp . kurstaki strain ABTS351 (MRID 41443401). 8

Based on the following reasoning, EPA agreed that no further testing was necessary to address this data requirement: (1) the results of the acute pulmonary toxicity /pathogenicity and acute injection toxicity/pathogenicity a nd acute oral toxicity studies s how that toxicity, pathogenicity and/or infectivity in mammals is not expected; (2) the scientific bridging argument provided by the applicant that the acute oral toxicity/pathoge nicity conducted with DiPel Technical showed no evidence of toxicity or pathogenicity, and it further showed that to tal clearance would occur by 47 days. For these reasons, it would be unnecessa ry to conduct additional studies to evaluate acute oral toxicity/pathogenicity.

Acute Pulmonary Toxicity/Pathogenicity – Ra t (Harmonized Guideline 885.3150); Master Record Identification Number (MRID No). 48559210: VBC-60225 containing the spores of Bacillus thuringiensis , subsp. kurstaki strain VBTS-2546 (Technical Powder) was tested for toxicity and pathogenicity in male and female CD ® rats by observation of the effects following a single intratracheal administra tion of approximately 1.0 x 10 8 colony forming units (cfu) of viable technical grade test substance. The study duration wa s 190 days; organ weights were collected on days 7, 12, 28 and 35.

Within the study design parameters measured, VBC-60225 ( Bacillus thuringiensis , subsp.

kurstaki strain VBTS-2546) was not found to be pat hogenic or toxic at the dose administered by

the intratracheal route to male and female CD ® rats. Total clearance of viable test substance from target organs was estimated at approxi mately 179 to 189 days (males or females, respectively) from challenge, and the absence of clinical signs or tissue abnormalities indicate the test substance is non-pathoge nic via intratracheal route. This study was found acceptable.

Acute Injection Toxicity/P athogenicity (Intravenous)

– Rat (Harmonized Guideline 885.3200;

MRID No. 48559211): Bacillus thuringiensis , subsp. kurstaki strain VBTS-2546 technical powder was not found to be pathogeni c or toxic at a dose of 6.7 x 10 7 cfu by the intravenous

route to male and female CD ® rats. Clearance of viable test substance from target organs (e.g., spleen) was estimated to occur at approximately 5 00 days from challenge for males and females. The absence of clinical signs or tissue abnorma lities indicated that th e test substance is nonpathogenic. This study was found acceptable.

6 B.A. Gingras, 2011, Toxicity/ Pathogenicity Testing of VBC-60225 following Acute Intratracheal Challenge in rats, Report No. 2297 SN2 (submitted). 7 B.A. Gingras, 2011, Toxicity/ Pathogenicity Testing of VBC-60225 following Acute Intravenous Challenge in rats, Report No. 2297 SN1.Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 10 of 38

Hypersensitivity Incidents (Harmonized Guideline 885.3400) : EPA regulations require that hypersensitivity incidents, includ ing immediate-type and delayed- type reactions of humans and domestic animals, that occur duri ng the testing of the te chnical grade of the active ingredient or manufacturing-use product, or ar e otherwise known to the applican t, 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 Bacillus thuringiensis , subsp. kurstaki strain VBTS-2546 is not a virus (refer to test note #4 of 40 CFR § 158.2140(d)).

Acute Oral Toxicity

– Rat (Harmonized Guidel ine 870.1100; MRID No. 48559212) : An acute oral toxicity test was conducted with rats to determine the potential for Bacillus thuringiensis

, subsp. kurstaki strain VBTS-2546 to produce toxicity fr om a single dose via the oral route. A limit dose of 5,000 milligrams of Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 technical powder per kilogram of body weight (mg/kg bw) was administer ed to three healthy female rats by oral gavage. All animals survived exposure to the test substance, gained body weight and appeared active and healthy during the study. There were no signs of gross toxicity, adverse pharmacologic effects, or abnormal beha vior. No gross abnormalities were noted for any of the animals when necropsied at the conclu sion of the 14-day observation period. This study was found acceptable and Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 Technical

Powder was classified as Toxicity Category IV for oral toxicity

Acute Dermal Toxicity – Rat (Harmo nized Guideline 870.1200; MRID No. 48559213) : An acute dermal toxicity test was conducted with rats to determine the potential for Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 to produce toxicity fr om a single topical application. Five thousand mg of Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 t echnical powder per kg bw was applied to the skin of ten healthy rats for 24 hours. All animals survived exposure to the test substance and gained body weight during the study. Other than the dermal irritation (erythema) noted at the dose site for three out of five females and one male between Days 1 and 2, there were no other adverse clin ical findings recorded for any animal over the course of the study. No gross abnormalities were noted for any of the animals when necropsied at the

conclusion of the 14- day observation period. This study was found acceptable and Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 Technical Powder was classified as Toxicity Category IV for dermal toxicity.

Acute Inhalation Toxicity

– Rat (Harmonized Guideline 870.1300; MRID No. 48559214) : In this study, ten healthy rats (five/sex) were exposed (single dose) to th e test atmosphere for four hours via the inhalation (nose-only exposure) route. The gravimetric chamber contained a concentration of 5.16 mg/L Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 technical powder. Following exposure to the test atmos phere, all animals were hypoactive and exhibited irregular respiration, moist rales (abnormal respiratory sounds), red nasal discharge and/or ataxia. However, all animals recovered by Day 2 and app eared active and healthy for the remainder of the 14-day observation period. Although all rats lost body weight by Day 1, all animals showed a continued weight gain thereafter through Day 14. No gross abnormalities were noted for the animals when necropsied at the conc lusion of the 14-day observation period. This study was found acceptable and Bacillus thuringiensis , subsp. kurstaki strain VBTS-2546 TechnicalBacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 11 of 38

Powder was classified as Toxicity Category IV fo r inhalation toxicity.

Primary Eye Irritation – Rabbit (Harmonized Guideline 870.2400; MRID No. 48559215) : A primary eye irritation study was conducted with rabbits to determine the potential for Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 to produce irri tation from a single instillation via

the ocular route. Under the conditi ons of this study, the test substance is classified as mildly irritating to the eye. This study was found acceptable and Bacillus thuringiensis subsp. kurstaki

strain VBTS-2546 Technical Po wder was classified as Toxicity Category III for eye irritation.

Primary Dermal Irritation – Rabbit (Har monized Guideline 870.2500; MRID No. 48559216) : A primary skin irritation study was conducted with rabbits to determine the potential for Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 to produce i rritation after a single topical application. Under the conditions of this study, the test substance is classified as moderately irritating to the skin. This study was found acceptable and Bacillus thuringiensis subsp. kurstaki

strain VBTS-2546 Technical Po wder was classified as Toxicity Category III for dermal

irritation.

Skin Sensitization – Guinea Pig (Harm onized Guideline 870.2600; MRID No. 48559217) : A dermal sensitization study was conducted with guinea pigs to determine the potential for Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 to produce sens itization after repeated topical applications. Based on the results of this study, the test substance is not considered to be a

contact sensitizer. The positive response observed in the historical positive control validation study with alpha-Hexylcinnamaldehyde Technical (HCA ) validates the test system used in this study. This study was found acceptable and Bacillus thuringiensis subsp. kurstaki strain VBTS2546 Technical Powder was classified as not a dermal sensitizer.

c. Endocrine Disruptors

As required by the Administrator under the Federal Food, Drug, and Cosmetic Act (FFDCA) section 408(p), EPA has developed the Endocrine Disruptor Screening Program (EDSP) and has begun to implement the screening program that is to be used to test all pesticides to determine whether certain substances (including pesticide ac tive and other ingredients) may have an effect in humans or wildlife similar to an effect pr oduced by a “naturally occurring estrogen, or other

such endocrine effects as the Administrato r may designate.” FFD CA section 408(p)(4), authorizes the Administrator, by order, to ex empt from the requirements of the Endocrine Disruptor Screening Program a biologic substance or other substance if a determination is made that the substance is anticipated not to produce any effect in humans similar to an effect produced by a naturally occurring estrogenic su bstance. Between October 2009 and February 2010, EPA issued test orders/data call-ins for th e first group of 67 chemicals, which contain 58 pesticide active ingredients a nd nine inert ingredients. Bacillus thuringiensis is not among the group of 58 pesticide active ingredients on the in itial list to be screened under the EDSP.

The Agency believes that Bacillus thuringiensis likely is a substance th at would not produce any effect in humans similar to an effect produced by a naturally occurring estrogenic substance. As such, and pursuant to Section 408(p)(4), EPA will determine in the future whether it can exempt Bacillus thuringiensis from the requirements of the Secti on 408(p) EDSP. In the event theBacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 12 of 38

Agency does determine to exempt this substance from the EDSP, an order will be issued. For further information on the status of the EDSP, the policies and procedur es, the list of 67 chemicals, future lists, the test guidelines and the Tier 1 screening ba ttery, 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.” Sect ion 408(c)(2)(A)(ii) of FFD CA defines “safe” to mean that “there is a reasonable certainty that no harm will result from aggregate exposure to the pesticide chemical residue, incl uding all antic ipated dietary exposures a nd all other exposures for

which there is reliable information.” This includes exposure through drinking water and in residential settings but does not include occupa tional exposure. Pursuant to secti on 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 f actors set forth in sect ion 408(b)(2)(C) of FFDCA, which require EPA to give special consideratio n to exposure of infants and children to the pesticide chemical residue in establishing a tole rance 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....” Additionally, section 408(b)(2)(D) of FFDCA requires that EPA consider “available information concerning the cumulative effects of [a particular pesticide’s] . . . residues and other substances that have a common mechanism of toxicity.”

EPA performs a number of analyses to determin e 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 throu gh other exposures that occur as a result of pesticide use in residential settings.

Consistent with section 408(b)(2 )(D) of FFDCA, EPA reviewed the available scientific data and other relevant information, and considered its validity, completeness, and reliability and the relationship of this information to human ris k. 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/pat hogenicity data and informati on discussed previously and presented in Table 3 in Appendix A , the data required for a FFDCA risk assessment for Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 have been fulfilled.

a. Aggregate Exposure

In examining aggregate exposure, section 408 of FFDCA directs EPA to consider available information concerning exposures from the pe sticide residue in food and all other nonoccupational exposures, includi ng drinking water from ground water or surface water and exposure through pesticide use in gardens, re sidential and other lawns and landscapes, or buildings (residential and ot her indoor uses). The exposure potential for the two end use products, VBC-60236 and Foray 2WG are discussed separately below.Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 13 of 38

VBC-60236 is a dry flowable insectic ide containing 67% active ingredient Bacillus thuringiensis subsp. kurstaki strain VBTS-2546. Valent BioSciences Cor poration intends to register this new product for use on agricultural crops grown in greenhouses/shade houses and outdoor nursery, fields, and ornamental trees; this product may be applied to plant foliage and fruit surfaces. VBC-60236 may be tank mixed with other labeled insecticides to e nhance control. Additions of other commonly used herbicides, insecticides, and fungi cides are generally not deleterious to VBC-60236. However, it should not be applied as a tank mix with, or within two days of application of alkaline products such as foliar nutrients, liqui d fertilizers, or metal-based fungicides ( e.g . copper sulfate-based materi als). Application methods include aerial, ground and chemigation.

Foray 2WG is a water dispersi ble granule insecticide cont aining 67% active ingredient Bacillus thuringiensis subsp. kurstaki strain VBTS-2546. Va lent BioSciences Corporation intends to register this new produc t for forestry uses, shade trees, or namentals, sugar maple trees, and ornamental fruits, nuts and citrus trees. App lication methods for Foray 2WG include aerial and ground sprays.

Food Exposure and Risk Characterization : Exposure to residues of Bacillus thuringiensis strain VBC-60236 in food may occur through the consum ption of raw agricultural commodities that have been treated with the end use product VBC-60236 Biological Insecticide Dry Flowable. EPA expects the a exposu re to food residues of Bacillus thuringiensis strain VBC-60236 to be negligible when compared to the ordinary, ever yday levels to which individuals are already exposed in the natural environment— Bacillus thuringiensis is commonly found in soil and on plant undergrowth. Food exposure is expected to be further reduced for the following reasons: (1) the proposed pesticide products will be dilu ted prior to applicati on and (2) rainfall and irrigation will further wash residues of the pesticide from the treate d crops. 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 lack of toxicity and path ogenicity demonstrated in the submitted studies.

Foray 2WG is not labeled for application to food cr ops, and thus its use is not expected to result

in residues in food.

Drinking Water Exposure and Risk Characterization : Residues of VBC-60236 and Foray 2WG may migrate to groundwater through runoff from treated fields, fore st and other use sites and to surface water through runoff and spray drift. The proposed directions for use specify dilution prior to application and use in chemigati on. These measures will reduce the amount of Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 in runoff into surface or percolation to ground

water.

Application may be made to ornamental ponds that are confined, and no runoff is expected from

this use.

If, following application to the sites listed on the V BC-60236 and Foray 2WG labels, residues of Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 are transf erred to surface or ground

waters that are intended for eventual human consumption an d subjected to sanitation (e.g.,Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 14 of 38

chlorination, pH adjustments, filtration, high te mperatures) in drinking water treatment plants, the residues would likely be removed from the fini shed drinking water. In the unlikely event that any residues of the pesticide occur in drinki ng 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 stud ies (refer to section IV.B.1.a. of this BRAD).

Non-occupational, Residential Ex posure and Risk Characterization : VBC-60236 is an agricultural-use pestic ide; it is not intended to be used in and around the home, or in schools, day

care facilities or other su ch settings, and its use is not e xpected to result in non-occupational residential exposure. Foray 2WG is labeled for us e in forests and on ornamentals, interiorscapes, golf courses, and lawns. Non- occupational residential exposur e may occur though contact with treated plants. EPA expects the aggregate exposur e to residues of Foray 2WG to be negligible when compared to the ordinary, everyday levels to which individua ls are already exposed in the natural environment, since Bacillus thuringiensis is commonly found in soil and on plant undergrowth. In the unlikely event that any re sidues 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 subm itted 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, when considering whether to establish, modify, or revoke a tolerance exemption, EPA consid er “available information concerning the cumulative effects of [a particular pesticide’s] . . . residues and other substances that have a common mechanism of toxicity.”

Bacillus thuringiensis is not known to have a common mech anism of toxicity with other substances; therefore, cumulative effects wi th other substances are not anticipated. For information regarding EPA’s efforts to identif y chemicals that may have a common mechanism of toxicity and to evaluate the cumulative eff ects of such chemicals, see EPA’s website at http://www.epa.gov/pesticides/cumulative

.

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

FFDCA section 408(b)(2)(C) provides that, in considering the esta blishment of a tolerance or tolerance exemption for a pesticide chemical re sidue, EPA shall assess th e available information about consumption patterns among infants and ch ildren, special susceptib ility of infants and children to pesticide chemical residues, and the cumulative effects on infa nts 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 tenf old (10X) margin of exposure (safety) for infants and children in the ca se of threshold effects to account for prenatal and postnatal toxicity and the completeness of th e database on toxicity and exposure unless EPA determines that a different margin of exposure (saf ety) will be safe for infants and children. This additional margin of exposure (safety) is comm only 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 reliab le data available to EPA support the choice of aBacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 15 of 38

different factor.

Based on its analyses of the submitted toxicity da ta (refer to section IV.B.1.a. and Table 3 in Appendix A ), EPA concludes that there are no threshol d effects of concern to infants, children,

or adults when Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 is used as labeled in accordance with good agricultural practices. The t oxicity of VBC-60236 and Foray 2WG is low and no toxicological effects or endpoints were identified. The Agency does not expect that infants and children will have special sensitivity to residues of VBC-60236 or Foray 2WG. 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 additi onal margin of exposure (safety) will be safe

for infants and children.

Based on the same data and EPA analysis presen ted directly above, the Agency is able to

conclude that there is a reasona ble certainty that no harm w ill result to the U.S. population, including infants and children, from a ggregate exposure to the residues of Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 when it is used as labeled and in accordance with good agricultural practices as an inse cticide. Such exposure includes all anticipated dietary exposures and all other exposures for which there is reliab le information. EPA arrived at this conclusion because, considered collectively, the data available on Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 do not demonstrate toxic po tential to mammals, including infants and

children.

3. Occupational Exposure and Risk Characterization

The available data (e.g., lack of toxicity noted for oral, dermal, and inhalation routes of exposure

to Bacillus thuringiensis subsp. kurstaki strain VBTS-2546) do not demo nstrate toxic potential to persons occupationally exposed to th is 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.

4. Human Health Risk Characterization

EPA considered human exposure to Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 in light of the standard for regi stration in FIFRA and the releva nt safety factors in FFDCA. A

determination has been made that no unreasonabl e adverse effects to the U.S. population in general, and to infants and children in particular, will result when Bacillus thuringiensis subsp.

kurstaki strain VBTS-2546 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 l iterature) submitted by th e applicant to support requests to waive nontarge t organism testing for Bacillus thuringiensis subsp. kurstaki strainBacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 16 of 38

VBTS-2546 are sufficient to fulfill the relevant microbial pesticide 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. 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 fr om the proposed pesticidal uses of Bacillus thuringiensis subsp. kurstaki strain VBTS-2546. Moreover, EPA has made a “No Effect”

determination for direct and indir ect effects to listed species and their designated critical habitats resulting from these same proposed pesticidal uses.

For a summary of the generic data requirements de scribed 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. 48559218): An acute avian oral toxicity study with Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 at the maximum

hazard dose showed that the acute oral LD 50 is > 1.83 x 10 9 cfu/kg of body weight per day. This study was acceptable.

Avian Inhalation Toxicity Pathogenicity (Harmonized Guideline 885.4100) (MRID No. 48559236): A scientific rationale was sufficien t to satisfy the data requirement. Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 is not considered toxic or pathogenic to birds. This rationale was acceptable.

Wild Mammals Toxicity/Pathogenicity (Harmonized Guideline 885.4150) (MRID No.

48559237): Tests required by 40 CFR § 158.2140 are adequa te and appropriate for assessment of

hazards to wild mammals. Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 showed no adverse toxicity or pathogenicity in rats e xposed via pulmonary and IV route (MRIDs 485592-10 and -11), or in toxicity tes ting (MRIDs 485592-12 through -13).

Nontarget Insects (Harmonized Guideline 885.4340) (MRID Nos. 48559222 , 48559223 and 48559221) : No studies with nontarget insect s were specifically provided for Bacillus thuringiensis subsp. kurstaki Strain VBTS-2546; however, two st udies with the registered DiPel HD-1 containing Bacillus thuringiensis subsp. kurstaki strain ABTS-351 were submitted, with rationale justifying bridging to studies with the registered Btk strain ABTS-351 and a review paper on effects of Bacillus thuringiensis on nontarget insects (Melin and Cozzi 1990). The two

submitted studies were conducted on an aphid parasitoid ( Aphidius rhopalosophi ) and a predatory mite ( Typhlodromus pyri ), which were exposed at 3.9 kg/ha. It is not possible to determine whether this exposure is a maximum h azard dose, given the current information about

the test material and that la bels for products containing the Bacillus thuringiensis subsp. kurstaki strain ABTS-351 have higher application rates. Although these studies were determined to be supplemental, they are acceptabl e for the risk assessment of Bacillus thuringiensis subsp.

kurstaki strain VBTS-2546.

Since the bridging rationale is sufficient to bridge to previously submitted data on Bacillus thuringiensis subsp. kurstaki strain ABTS-351, previous conclusions regarding Btk ABTS-351Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 17 of 38

are applicable to this risk assessment for Bacillus thuringiensis subsp. kurstaki strain VBTS2546. An evaluation of the ABTS-351 strain was included in the Bacillus thuringiensis Reregistraion Eligibility Decision (USEPA 1998). This evaluation concluded that Bt subsp.

kurstaki displays activity toward Lepidopteran insect s and some other species, and may result in temporary reductions in susceptible insect popu lations; however, the risk to nontarget insect populations is expected to be minimal for labele d uses as long as the product does not contain β

exotoxins. The bridging rationale and a paper from scientific literature were determined acceptable.

Honey Bees (Harmonized Guideline 885.4380) (MRID No. 48559224) : Valent also submitted a study with honey bees ( Apis mellifera ) exposed to Bacillus thuringiensis subsp. kurstaki strain VBTS-2546. Based on this study, th e acute oral and contact LC 50 was determined to be > 100 mg a.i./bee; however, the study was supplemental b ecause it was only 48 hours in duration. Studies

with Bt subsp. kurstaki submitted previously to EPA did not show significant adverse effects to honey bees (1998 Bt Reregistration Eligibility Decision), a nd concerns for these insects have not been raised and are not expected with this new a.i.

Nontarget Plants (Harmonized Guideline 885.4300); (MRID No. 48559221) : Scientific and bridging rationales were submitted to satisfy th e data requirement for nontarget plants. The microorganism Bacillus thuringiensis is well characterized, is not taxonomically related to known plant pathogens, and pe sticide products containing Bt have had a history of safe use in the U.S. Therefore, adverse effects on nontarget plants are not expected to result from the proposed applications of Bacillus thuringiensis subsp. kurstaki strain VBTS-2546. Submitted bridging

rationale was determined not to be critical to th e risk assessment for plants because no data are available for bridging. Studies with nontarge t plants have typically been waived for Bacillus thuringiensis . The scientific rati onale was acceptable.

b. Aquatic Animals and Plants

Freshwater Fish (Harmonized Guideline 885.4200) (MRID No. 48559219): Valent submitted rationale to justify bridging to studies on freshwater fish that were performed on Bacillus thuringiensis subsp. kurstaki ABTS-351. The rationale specifical ly cited studies with rainbow trout ( Oncorhynchus mykiss ; MRID 41443406) and bluegill ( Lepomis macrochirus ; MRID 41443405), which were included the Bt RED. The LC 50 was >1.5 x 10 10 cfu/L for rainbow trout, and this study was classified as supplemental, since it was unclear whether the mortalities observed were due to the physical effects resultin g from high turbidity and suspended solids. The study with bluegill was classified as core (acceptable) and the LC 50 was >1.5 x 10 10 cfu/L for this species. These studies were conducted at exposure levels above those that would be expected from labeled applications of Bacillus thuringiensis subsp. kurstaki strain VBTS-2546; therefore,

adverse effects to freshwater fi sh are not expected as a result of the proposed applications.

Freshwater Invertebrates (Harmoni zed Guideline 885.4240) (MRID No. 48559220): A study

with Daphnia magna was submitted, in which Daphnia were exposed to water containing the technical product Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 for 21 days. The 21- day EC 50 , based on adult immobilit y, was 21 mg/L (5.7 x 10 6 cfu/mL) and EC 50 based on reproduction was 24 mg/L (6.5 x 10 6 cfu/mL). The NOEC value, based on mortality was 10Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 18 of 38

mg/L (2.7 x 10 6 cfu/mL). Given the percentage of the a.i. in the EPs and the highest labeled application rate of 1 lb/acre, a direct applic ation to a body of water equivalent to the EPA Standard Wetland would be approximately 0.5 mg/L, which is 1/20 the NOEC value. Additionally, mortalities observed at the higher concentrations ma y be due to physical effects related to excessive particles in the water. Mort ality in the filtered trea tment prepared with the same amounts as the highest treatment levels was 60%, as compared to 100%. Since exposure resulting from the proposed applications is we ll below the defined toxicity endpoints, adverse

effects to freshwater invertebrates are not expected. This st udy is acceptable.

Estuarine/Marine Fish and Invertebrates (Harmonized Guideline 885.4280), (MRID No.

48559238): To fulfill the data requirement for marine/estuarine fish and invertebrate testing the applicant submitted a rationale bridging to cited studies with Bacillus thuringiensis subsp.

kurstaki strain ABTS-351. The studies cited show th at the ABTS-351 strain is not toxic to marine/estuarine invertebrates, and both of thes e studies were classified as core (acceptable). Specifically, the NOEL was >2.9 x 10 9 cfu/g of food for grass shrimp ( Palaemonetes vulgaris

, MRID 41540802) and was 500 mg/kg (1 x 10 7 cfu/g sediment) for the benthic copepod ( Amphiascus minutus , MRID 41443408). Although no fish studi es were specifically cited, a study with the ABTS-351 strain on sheepshead minnow ( Cyprinodon variegates ; MRID 41540801) is available for use in this ri sk assessment. In this study, the LC 50 was >2.9 x 10 10

cfu/L in water and >2.9 x 10 9 cfu/g food over a 30-day expos ure period. Exposure in marine environments is expected to be lower than th ese levels (maximum hazard dosing is at least 10 6

cfu/mL); therefore, the proposed applications of Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 are not expected to result in adverse effects to ma rine or estuarine fish or invertebrates. This bridgi ng rationale is acceptable.

2. Environmental Fate Assessment Data

Environmental fate and groundwat er data requirements (Tier II testing) were not triggered because there were no significant adverse effects to nontarget organisms in the Tier I toxicity studies, and because of acceptable rationale to address certain nontarget organism data requirements not fulfilled by testing, as discus sed in C.1. above and summarized in Table 4, Appendix A.

3. Ecological Exposure and Risk Characterization

The end use product (EP), VBC-60236 Biological In secticide Dry Flowable, is proposed for application to many types of field crops, in cluding fruit and nut trees, watercress (when no standing water is present), greenhouse/shade house and outdoor nurse ry plants, aquatic ornamentals (self-contained, cl osed-system ornamental ponds only), and ornamental trees. Proposed application methods include ground an d aerial sprays, chemigation, and direct application to water (for aqua tic ornamentals only). The EP, Fo ray 2WG Biological Insecticide, is proposed for use on forests, shade trees, ornament als, sugar maple trees, and ornamental fruit, nut, and citrus trees, and is intended to be applied by ground or aerial spray. Since the proposed applications are expected to resu lt in contact of the a. i. with foliar surfaces and soil, and spray drift is expected to o ccur, exposure to terrestrial and aqua tic nontarget organisms is possible. However, based on the lack of toxicity shown in studies and rationale discussed above, adverseBacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 19 of 38

effects to nontarget species ar e not anticipated as a result of the proposed labeled uses.

4. Endangered Species Assessment

EPA has determined that no effects are antici pated for most nontarge t species exposed to Bacillus thuringiensis subsp. kurstaki , Strain VBTS-2546 as a result of the proposed labeled applications, including mammals, birds, terres trial and aquatic plants , and freshwater and marine/estuarine animals. Although sustained popul ation-level effects to nontarget insects are not expected, it is known that Bt subsp. kurstaki can affect certain speci es of insects (e.g., lepidopterans). Therefore, a “No Ef fect” determination is made for direct and indirect effects to federally listed endangered and threatened (“list ed”) mammals, birds, te rrestrial and aquatic plants, and freshwater and marine /estuarine animals and for thei r designated critical habitat.

Direct effects to indivi duals of listed insect sp ecies cannot be precluded, nor can indirect effects to listed species that obligatorily depend on speci fic listed insects. Additional analysis of the potential effects to the listed taxa is necessa ry to make a determination for these species.

V. ENVIRONMENTAL JUSTICE

EPA seeks to achieve environmen tal justice—the fair treatment and meaningful involvement of all people regardless of race, color, national or igin, or income—with respect to the development, implementation, and enforcement of environmen tal laws, regulations, and policies. Fair treatment means that no group of people, includi ng racial, ethnic, or socioeconomic groups, should bear a disproportionate sh are of the negative environmenta l consequences resulting from industrial, municipal, and commercial operations or the execution of federal, state, local, and tribal environmental programs and policies. Mean ingful involvement mean s that (1) potentially affected community residents have an appropriate opportunity to participat e in decisions about a proposed activity that will aff ect their environment and/or heal th; (2) the public’s contribution can influence the regulatory agency’s decision; (3) the concerns of all participants involved will be considered in the decision-making process; and (4) the decision-makers seek out and facilitate the involvement of those potentially affected. EPA has this goal for all communities and persons

across the U.S.

To help address potential envir onmental justice issues, EPA seeks information on any groups or segments of the population who, as a result their location, cultural practic es, or other factors, may have atypical, unusually high exposure to Bacillus thuringiensis subsp. kurstaki strain VBTS-2546, compared to the general populati on. Please comment if you are aware of any subpopulations that may have atypical, unusua lly high exposure compared to the general population.

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

.

VI. RISK MANAGEMENT DECISION

Section 3(c)(5) of FIFRA provides for the regist ration of a pesticide pr ovided that all the following determinations are made: (1) its composition is such as to warrant the proposed claimsBacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 20 of 38

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 unreasona ble adverse effects on the environment.

To satisfy criterion 1, the compositions of Bacillus thuringiensis subsp. kurstaki strain VBTS2546 pesticide products have well-known prope rties. EPA has no knowledge that would

contradict the claims made on the labels fo r the manufacturing-use pesticide product, VBC60219 Biological Insecticide Sl urry ,VBC-60225 Biological Ins ecticide Technical Powder Active Ingredient and/or two end-use pesticide products, VBC- 60236 Biological Insecticide Dry Flowable and Foray 2WG Biologi cal Insecticide, and such pesticide products are not expected

to cause unreasonable adverse effects on the environment when used according to their respective label instructions. Cr iterion 2 is satisfied by the cu rrent proposed product labels, as well as the data and information presented in this document. It is believed that Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 pesticide products will not cause any

unreasonable adverse effects on the environmen t, and the two EPs, VBC-60236 Biological Insecticide Dry Flowable and Foray 2WG Biologi cal Insecticide in particular, are likely to control the caterpillar pests de scribed on the product labels, sa tisfying criterion 3. Criterion 4 is

satisfied in that Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 pe sticide products are not expected to cause unreasonable adverse effe cts when used according to label instructions. Therefore, VBC-60219 Biologica l Insecticide Slurry ,VBC- 60225 Biological Insecticide Technical Powder Active Ingredient and the two end-use pesticide products, VBC-60236 Biological Insecticide Dry Flowable and Fo ray 2WG Biological Ins ecticide, containing Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 as a new activ e ingredient, are eligible for registration under FIFRA section 3(c)(5) for the labeled uses.

VII. ACTIONS REQUIRED OF THE REGISTRANT

A. Final Printed Labeling

Before releasing pestic ide products containing Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 for shipment, the regist rant is required to provide a ppropriate final printed labeling

to EPA.

B. Terms of Registration

As terms of the registrations, the applicant must submit the results of storage stability (Harmonized Guideline 830.6317) and corrosion characteristics (Harmonized Guideline 830.6320) studies within one year of registration.

C. Reporting of Adverse Effects and Hypersensitivity Incidents

Notwithstanding the information stated in the prev ious sections, it should be clearly understood that certain specific data are requi red to be reported to EPA as a requirement for maintaining the federal registration for a pesticide product. A br ief summary of these t ypes of data are listed

below.Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 21 of 38

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 co nfirmed 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 Registrati on of Pesticide Products Containing Bacillus thuringiensis subsp. kurstaki strain VBTS-2546.

MRID Citation

41443401 Carter, N. (1993) Acute oral toxicity and infectivity/pa thogenicity to rats of ABG 6345, Report no. G-7239.222

48655601 Bujor, D. (2011) VBC-60219: Product An alysis and Physical and Chemical Properties: Waiver Request. Proj ect Number: VBC/60219/W/1. Unpublished study prepared by Valent BioSciences Corporation. 7p. 48655602 Rowell, R. (2011) Method of Production and Quality Control for VBC-60219, (Bacillus thuringiensis subsp. kurstaki, Strain VBTS-2546 Slurry Concentrate): Replacing MRID 48559203. Project Numb er: VBC/60219/MP/1. Unpublished study prepared by Abbott Laboratories. 149p. 48655603 Bujor, D. (2011) VBC-60219: Acute Oral Toxicity: Waiver Request. Project Number: VBC/60219/W/2. Unpub lished study prepared by Valent BioSciences Corporation. 4p. 48655604 Bujor, D. (2011) VBC-60219: Acute Derm al Toxicity: Waiver Request. Project Number: VBC/60219/W/3. Unpub lished study prepared by Valent BioSciences Corporation. 4p. 48655605 Bujor, D. (2011) VBC-60219: Acute Inhala tion Toxicity: Waiver Request. Project Number: VBC/60219/W/4. Unpub lished study prepared by Valent BioSciences Corporation. 4p. 48655606 Bujor, D. (2011) VBC-60219: Acute Ey e Irritation: Waiver Request. Project Number: VBC/60219/W/5. Unpub lished study prepared by Valent BioSciences Corporation. 4p. 48655607 Bujor, D. (2011) VBC-60219: Primary Derm al Irritation: Waiver Request. Project Number: VBC/60219/W/6. Unpub lished study prepared by Valent BioSciences Corporation. 4p. 48677401 Bujor, D. (2011) VBC-60219: Hypersensi tivity Statement. Project Number: VBC/60219/H1. Unpublished study prepared by Valent BioSciences Corporation. 4p. 48677402 Bujor, D. (2011) VBC-60225: Product An alysis and Physical and Chemical Properties: Waiver Request. Proj ect Number: VBC/60225/W/1. Unpublished study prepared by Valent BioSciences Corporation. 13p. 48677403 Arndt, T.; Marin, J. (2011) Physical Properties of VBC-60236 Upon Storage atBacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 22 of 38

30(Degrees Celsius) for 18 Weeks. Project Number: 2152W. Unpublished study prepared by PTRL West, Inc., Abbot t Laboratories and Accugenix. 87p. 48677404 Bujor, D. (2011) VBC-60225: Hypersensi tivity Statement. Project Number: VBC/60225/H1. Unpublished study prepared by Valent BioSciences Corporation. 4p. 48677405 Bujor, D. (2011) VBC-60236: Hypersensi tivity Statement. Project Number: VBC/60236/H1. Unpublished study prepared by Valent BioSciences Corporation. 4p. 48559218 Hubbard, P.; Beavers, J. (2011) Bacillus thuringiensis, subsp. kurstaki, (Btk),

Strain VBTS-2546: An Avian Oral Pat hogenicity and Toxicity Study in the Northern Bobwhite. Project Number: 529/124. Unpublished study prepared by Wildlife International, Ltd. 52p. 48559219 Bujor, D. (2011) VBC-60225: Freshwater Fish Toxicity/Pathogenicity: Request for No Further Testing. Project Number: VBTS/2546/R/2. Unpublished study prepared by Valent BioS ciences Corporation. 6p. 48559220 Gallagher, S.; Krueger, H. (2011) Bacillus thuringiensis, subsp. kurstaki, (Btk), Strain VBTS-2546: A 21-Day Life Cycle T oxicity and Pathogeni city Test with the Cladoceran (Daphnia magna): Fina l Report. Project Number: 592A/120. Unpublished study prepared by Wildlife International, Ltd. 77p. 48559221 Herrero, M ; Bujor, D. (2011) VBC-60225: Nontarget Insect Testing: Request for no further Testing. Project Number: VB TS/2546/R/4. Unpublished study prepared by Valent BioSciences Corporation. 25p. 48559222 Warmers, C. (2005) DiPel Technical Powder: Acute Toxicity to the Aphid Parasitoid. Aphidius rhopalo siphi De Stefani Perez (Hym enoptera, Braconidae) in the Laboratory (Limit Test): Final Report. Project Number: 20051212/01/NLAP. Unpublished study prepared by GAB Biotechnologie Gmbh. 30p. 48559224 Verge, E. (2010) VBC-60225 - Acute Oral and Contact Toxicity to the Honeybee Apis mellifera L. in the Laboratory: Draft Report. Project Number: VBC/60225 S10/00883 S10/00883/L1/BLEU. Unpublished st udy prepared by Eurofins - GAB GmbH. 28p. 48559236 Bujor, D. (2011) VBC-60225: Avian Inha lation Toxicity/Pat hogenicity: Waiver Request. Project Number: VBTS/2546/W/ 1. Unpublished study prepared by Valent BioSciences Corporation. 4p. 48559237 Bujor, D. (2011) VBC-60225: Wild Mammal Toxicity/Pathogenicity: Request for No Further Testing. Project Number : VBTS/2546/W/2. Unpublished study prepared by Valent BioS ciences Corporation. 5p. 48559238 Bujor, D. (2011) VBC-60225: Estuarine/Ma rine Fish and Inve rtebrate Testing: Waiver Request. Project Number: VB TS/2546/W/3. Unpublished study prepared by Valent BioSciences Corporation. 4p. 48559239 Herrero, M.; Bujor, D. (2011) VBC- 60225: Nontarget Plant Testing. Project

Number: VBTS/2546/W/4. U npublished study prepared by Valent BioSciences Corporation. 4p.

B. Environmental Protection Agency Risk Assessment Memoranda

U.S. EPA. 2012a. Environmental risk assessm ent for the FIFRA Section 3 registration ofBacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 23 of 38

Bacillus thuringiensis subsp. kurstaki strain VBTS-2546. Memorandum from S.Borges. to A. Sibold draft dated August. 17, 2012.

U.S. EPA. 2011b Review of product chemistry, manu facturing process and ac ute toxicity studies for Section 3 registration of the end use products VBC-60236 and Foray 2WG (EPA Reg. No.73049-UTO) containing the TGAI VBC-60225 ( Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 Technical Powder); Memorandum from J. Facey, Ph.D. and J.L. Kough, Ph.D. to A. Sibold draft dated August 22, 2012.

U.S. EPA. 2004. Drinking Water Treatment. U.S. Environmental Protection Agency. Office of

Water, EPA 816-F-04-034. 2004. http://water.epa.gov/lawsregs/guidance/ sdwa/upload/2009_08_28_sdwa_fs_30ann_treatment _web.pdf

U.S. EPA. 1998. Reregistration Eligibility Deci sion (RED): Bacillus thuringiensis. USEPA Office of Prevention, Pesticides, and Toxi c Substances, Washington, DC. EPA 738-R-98-

004.

U.S. EPA. 1995. Pesticide Registration (PR) Notice 95-3: Reduction of Worker protection Standard (WPS) Interim Restricted Entry Interv als (REIs) for Certain Low Risk Pesticides.

June 7, 1995.

C. Other References

Benson, T., (2011), Microbial Characterization of Bacillus thuringiensis subsp. kurstaki, Strain VBTS-2546, Report N o. VBTS-2546-2011-1.

Gingras, B.A., (2011), Toxicity/ Pathogeni city Testing of VBC-60225 following Acute Intratracheal Challenge in rats, Report No. 2297 SN2.

Gingras, B.A., (2011), Toxicity/ Pathogeni city Testing of VBC-60225 following Acute Intravenous Challenge in rats, Report No. 2297 SN1.

Green, M., M. Heumann, R. Sokolow, L.R. Fost er, R. Bryant, M.Skeels, Public Health Implication of the Microbial Pe sticide Bacillus th uringiensis: An Epidemiological Study, Oregon, 1985-89, American Journal of Pub lic Health, July 1990, Vol. 80, No. 7.

Hill et al. (2004) Fluorescent amplified fragment length polymorphism of Bacillus anthracis

, Bacillus cereus , and Bacillus thuringiensis isolates. Appl. Environ. Microbiol . 70(2): 1068-1080.

LANL Report: “Amplified Fragment Length Polymo rphism (AFLP) Analysis of two Valent

Biosciences Bacillus thuringiensis Strains” LANL Project FIA-10-029.

Letowski et al. (2005) Assessment of cry1 gene contents of Bacillus thuringiensis strains by use of DNA microarrays. Appl. Environ. Microbiol . 71 (9): 5391-5398.Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 24 of 38

Lowe, C., 2011. VBC-60225: Acute Oral Toxicity Up and Down Procedure in rats, Report No.

31269.

Melin, B.E. and E.M. Cozzi. 1990. Safety to nontar get invertebrates of Le pidopteran strains of Bacillus thuringiensis and their β -exotoxins. Pp. 149-167 In Laird, M., Lacey, L.A. & Davidson, E.W.D., Eds., Safety of Microbial Insecticides . CRC Press, Boca Raton.

Noble, M. A., P.D. Riben, G.J.Cook, Microbial and Epidemiological Surveillance Programme to

Monitor the Health Effects of Foray 48B BTK Spray, British Columbia, September 30, 1992.Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document Page 25 of 38

This page intentionally left blank.APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS(40 CFR PART 158– SUBPART V)TABLE 1. Product Analysis Data Requirements forBacillus thuringiensissubsp.kurstakistrain VBTS-2546, includingVBC-60219 Biological Insecticide Slurry, VBC- 60225 Biological Insecticide (Technical Powder Active Ingredient)and Two End-Use Pesticide Products (EPs), Foray 2WG and VBC-60236B Biological Insecticide Dry Flowable (40 CFR § 158.2120)HarmonizedGuidelineNumberData RequirementResultsMRID No.Slurry TGAI EPs885.1100Product IdentitySubmitted data fulfill therequirement for product identity.VBC-60219 BiologicalInsecticide Slurry contains 16%by weightBacillusthuringiensissubsp.kurstakistrain VBTS-2546fermentation solids, spores,and insecticidal toxins.Submitted data fulfill therequirement for product identity.VBC-60225 BiologicalInsecticide Technical Powdercontains 98% by weightBacillus thuringiensissubsp.kurstakistrainVBTS-2546 fermentationsolids, spores, andinsecticidal toxinsSubmitted data fulfill the requirementfor product identity. The EPsForay2WG and VBC-60236Bcontain67%Bacillus thuringiensissubsp.kurstakistrain VBTS-2546fermentation solids, spores, andinsecticidal toxins48559203486556024855920548559210885.1200Manufacturing ProcessSubmitted data fulfill the requirement for manufacturing process.885.1300Discussion of Formationof UnintentionalIngredientsSubmitted data fulfill the requirement for discussion of formation of unintentional ingredients.N/ADeposition of a Samplein a NationallyRecognized CultureCollectionA sample ofBacillus thuringiensissubsp.kurstakistrainVBTS-2546is on deposit with the American TypeCulture Collection under Accession Number ATCC SD-6283885.1400 Analysis of Samples Rationale submitted in supportof a request to waive this datarequirement was foundacceptable.As a term of VBC-60225Biological Insectide TGAIregistration, EPA is requiringsubmission of a storage stabilitystudy within one year ofregistration.Submitted data fulfill therequirement for analysis ofsamples.885.1500 Certification of LimitsLimits are adequate/acceptable.Bacillus thuringiensissubsp.kurstakistrain VBTS-2546Biopesticides Registration Action DocumentPage 27 of 38Bacillus thuringiensissubsp.kurstakistrain VBTS-2546Biopesticides Registration Action DocumentPage 28 of 38TABLE 2. Physical and Chemical CharacteristicsforBacillus thuringiensissubsp.kurstakistrain VBTS-2546, includingVBC-60219 Biological Insecticide Slurry, VBC- 60225 Biological Insecticide (Technical Powder Active Ingredient)and Two End-Use Pesticide Products (EPs), Foray 2WG and VBC-60236B Biological Insecticide Dry Flowable(40 CFR § 158.2120)HarmonizedGuidelineNumberData RequirementResultsMRID No.Slurry TGAI EPs830.63021Color Light brown Yellow-brown Not required for EP48559203485592054855922548655602830.63031Physical State Slurry at 20°C Solid (fine powder) at 20°C Solid830.63041Odor No discernible odor No discernible odor Not required for EP830.63131Stability to Normal andElevatedTemperatures, Metals,and Metal IonsRationale submitted in support ofa request to waive this datarequirement was foundacceptable.This TGAI was prepared to be usedin toxicology and ecotoxicologystudies and is not used for any furtherformulations at this time. As a termof VBC-60225 Biological InsecticideTGAI registration, EPA is requiringsubmission of a storage stabilitystudy if the product is used toformulate end use products.Not required for EP830.6317 Storage StabilityRationale submitted in support ofa request to waive this datarequirement was foundacceptable.This TGAI was prepared to be usedin toxicology and ecotoxicologystudies and is not used for any furtherformulations at this time. As a termof VBC-60225 Biological InsecticideTGAI registration, EPA is requiringsubmission of a storage stabilitystudy if the product is used toformulate end use products.As a term of the registration, EPA isrequiring submission of a storage stabilitystudy within one year of registration.830.63192Miscibility Not requiredNot required because VBC-60225Biological Insectide TGAI is not anemulsifiable liquid form of amicrobial pesticide (refer to test note#2 of 40 CFR § 158.2120(d)).Not requiredBacillus thuringiensissubsp.kurstakistrain VBTS-2546Biopesticides Registration Action DocumentPage 29 of 38TABLE 2. Physical and Chemical CharacteristicsforBacillus thuringiensissubsp.kurstakistrain VBTS-2546, includingVBC-60219 Biological Insecticide Slurry, VBC- 60225 Biological Insecticide (Technical Powder Active Ingredient)and Two End-Use Pesticide Products (EPs), Foray 2WG and VBC-60236B Biological Insecticide Dry Flowable(40 CFR § 158.2120)HarmonizedGuidelineNumberData RequirementResultsMRID No.Slurry TGAI EPs830.6320CorrosionCharacteristicsRationale submitted in support ofa request to waive this datarequirement was foundacceptable.This TGAI was prepared to be usedin toxicology and ecotoxicologystudies and is not used for any furtherformulations at this time. As a termof VBC-60225 Biological InsectideTGAI registration, EPA is requiringsubmission of a corrosioncharacteristics study if the product isused to formulateend use products.As a term of VBC-60236 AND FORAY2WG registrations, EPA is requiringsubmission of storage stability andcorrosion characteristics studies withinone year of registration.830.70001pHThe pH of a 1% w/v dispersion ofVBC-60219 is 4.4.The pH of a 1% w/v dispersion ofVBC-60225 is 4.6.Not required for EP830.71003ViscosityPseudoplastic behavior; range 46-270 mPa.s@ 20°C and 50- 1700mPa.s@ 40°CNot required Not required830.7300Density/RelativeDensity/Bulk Density(Specific Gravity)1.04 1.39Pour density = 0.36 g/mlTap density = 0.42 g/ml1According to 40 CFR § 158.2120, these data are only required for the technical grade of the active ingredient.2According to 40 CFR § 158.2120, these data are only required for emulsifiable liquid forms of microbial pesticides.3According to 40 CFR § 158.2120, these data are only required for liquid forms of microbial pesticides.Bacillus thuringiensissubsp.kurstakistrain VBTS-2546Biopesticides Registration Action DocumentPage 30 of 38TABLE 3. ToxicologyData Requirements forBacillus thuringiensissubsp.kurstakistrain VBTS-2546, includingVBC-60219 Biological Insecticide Slurry, VBC- 60225 Biological Insecticide (Technical Powder Active Ingredient)and Two End-Use Pesticide Products (EPs), Foray 2WG and VBC-60236B Biological Insecticide Dry Flowable (40 CFR § 158.2140)Harmonized GuidelineNumberDataRequirementResultsMRID No.Slurry TGAI EPTier I885.30501Acute OralToxicity/PathogenicityNAGiven that acute pulmonarytoxicity pathogenicity, acuteinjection toxicity pathogenicityand acute oral toxicity testing wereconducted with the TGAI, toxicity,pathogenicity and/or infectivity inmammals is not expectedfollowing exposure to the TGAIvia the oral route. The applicantsubmitted scientific rationale tocompare Bacillus thuringiensissubsp. kurstaki strain VBTS-2546to previously registered strains. Itwould be inappropriate andunnecessary to conduct testingwith this active ingredient toevaluate these endpoints. Thus,EPA waived testing for thepathogenicity component of theacute oral toxicity/pathogenicitydata requirement.N/A48559201Bacillus thuringiensissubsp.kurstakistrain VBTS-2546Biopesticides Registration Action DocumentPage 31 of 38TABLE 3. ToxicologyData Requirements forBacillus thuringiensissubsp.kurstakistrain VBTS-2546, includingVBC-60219 Biological Insecticide Slurry, VBC- 60225 Biological Insecticide (Technical Powder Active Ingredient)and Two End-Use Pesticide Products (EPs), Foray 2WG and VBC-60236B Biological Insecticide Dry Flowable (40 CFR § 158.2140)Harmonized GuidelineNumberDataRequirementResultsMRID No.Slurry TGAI EP885.31501AcutePulmonaryToxicity/PathogenicityNAVBC-60225 (Bacillusthuringiensis, subsp. kurstaki,strain VBTS-2546) was notpathogenic, infective or toxic at adose of 1.0 x 108 colony formingunits (cfu) of viable technicalgrade (TG) test substance peranimal, when administered by theintratracheal route. Classification:AcceptableN/A48559210885.32001AcuteInjectionToxicity/Pathogenicity(Intravenous)NAVBC-60225 (Bacillusthuringiensis, subsp. kurstaki,strain VBTS-2546) was notpathogenic, infective or toxic at adose of 6.7 x 107 cfu whenadministered bythe intravenousroute.Classification: AcceptableN/A 48559211885.3400Hypersensitivity IncidentsHypersensitivity incidents, including immediate-type and delayed-type reactions of humans ordomestic animals, that occur duringthe testing or production of the slurry, TGAI, MP or EP, or areotherwise known to the applicant, must be reported if they occur (refer to test note #3 of 40 CFR §158.2140(d)).N/A885.3500 Cell CultureNot required because Bacillus thuringiensis subsp. kurstaki strainVBTS-2546is not a virus (refer totest note #4 of 40 CFR § 158.2140(d)).N/ABacillus thuringiensissubsp.kurstakistrain VBTS-2546Biopesticides Registration Action DocumentPage 32 of 38TABLE 3. ToxicologyData Requirements forBacillus thuringiensissubsp.kurstakistrain VBTS-2546, includingVBC-60219 Biological Insecticide Slurry, VBC- 60225 Biological Insecticide (Technical Powder Active Ingredient)and Two End-Use Pesticide Products (EPs), Foray 2WG and VBC-60236B Biological Insecticide Dry Flowable (40 CFR § 158.2140)Harmonized GuidelineNumberDataRequirementResultsMRID No.Slurry TGAI EP870.1100Acute OralToxicityAcute oral toxicity testingwas conducted with theTGAI and was foundacceptable, ToxicityCategory IV. Thecombination of inertingredients in this slurry istested in the TGAI and is notexpected to pose anysignificant human healthrisks via the oral route ofexposure. It would beinappropriate andunnecessary to conducttesting with this activeingredient to evaluate thisendpoint. Thus, EPA waivedtesting for the of the acuteoral toxicity datarequirement.TOXICITY CATEGORYIVOral LD50 female rats > 5,000mg/kg bodyweight (bw)Classification: AcceptableTOXICITY CATEGORY IVOral LD50 female rats>5000 mg/kg bwClassification: AcceptableToxicity Category IV4855921248559230Bacillus thuringiensissubsp.kurstakistrain VBTS-2546Biopesticides Registration Action DocumentPage 33 of 38TABLE 3. ToxicologyData Requirements forBacillus thuringiensissubsp.kurstakistrain VBTS-2546, includingVBC-60219 Biological Insecticide Slurry, VBC- 60225 Biological Insecticide (Technical Powder Active Ingredient)and Two End-Use Pesticide Products (EPs), Foray 2WG and VBC-60236B Biological Insecticide Dry Flowable (40 CFR § 158.2140)Harmonized GuidelineNumberDataRequirementResultsMRID No.Slurry TGAI EP870.1200Acute DermalToxicityAcute dermal toxicity testingwas conducted with theTGAI and was foundacceptable, ToxicityCategory IV. Thecombination of inertingredients in this slurry istested in the TGAI and is notexpected to pose anysignificant human healthrisks via the dermal route ofexposure. It would beinappropriate andunnecessary to conducttesting with this activeingredient to evaluate thisendpoint. Thus, EPA waivedtesting for the of the acutedermal toxicity datarequirement. TOXICITYCATEGORY IVDermal LD50 combined (male andfemale) rats > 5,000 mg/kgClassification: AcceptableTOXICITY CATEGORY IVDermal LD50 combined(male and female) rats >5,000 mg/kgClassification: AcceptableToxicity Category IV4855921348559231Bacillus thuringiensissubsp.kurstakistrain VBTS-2546Biopesticides Registration Action DocumentPage 34 of 38TABLE 3. ToxicologyData Requirements forBacillus thuringiensissubsp.kurstakistrain VBTS-2546, includingVBC-60219 Biological Insecticide Slurry, VBC- 60225 Biological Insecticide (Technical Powder Active Ingredient)and Two End-Use Pesticide Products (EPs), Foray 2WG and VBC-60236B Biological Insecticide Dry Flowable (40 CFR § 158.2140)Harmonized GuidelineNumberDataRequirementResultsMRID No.Slurry TGAI EP870.1300AcuteInhalationToxicityAcute inhalation toxicitytesting was conducted withthe TGAI and was foundacceptable, ToxicityCategory IV. Thecombination of inertingredients in this slurry istested in the TGAI and is notexpected to pose anysignificant human healthrisks via the inhalation routeof exposure. It would beinappropriate andunnecessary to conducttesting with this activeingredient to evaluate thisendpoint. Thus, EPA waivedtesting for the of the acuteinhalation toxicity datarequirement. TOXICITYCATEGORY IVInhalation LC50 combined (maleand female) rats > 5.16 mg/LClassification: AcceptableTOXICITY CATEGORY IVLC50 combined (male andfemale) rats >5.24 mg/LClassification: AcceptableToxicity Category IV4855921448559232Bacillus thuringiensissubsp.kurstakistrain VBTS-2546Biopesticides Registration Action DocumentPage 35 of 38TABLE 3. ToxicologyData Requirements forBacillus thuringiensissubsp.kurstakistrain VBTS-2546, includingVBC-60219 Biological Insecticide Slurry, VBC- 60225 Biological Insecticide (Technical Powder Active Ingredient)and Two End-Use Pesticide Products (EPs), Foray 2WG and VBC-60236B Biological Insecticide Dry Flowable (40 CFR § 158.2140)Harmonized GuidelineNumberDataRequirementResultsMRID No.Slurry TGAI EP870.2400Acute EyeIrritationAcute eye irritation testingwas conducted with theTGAI and was foundacceptable, ToxicityCategory III. Thecombination of inertingredients in this slurry istested in the TGAI and is notexpected to pose anyadditional human healthrisks via the eye route ofexposure. It would beinappropriate andunnecessary to conducttesting with this activeingredient to evaluate thisendpoint. Thus, EPA waivedtesting for the of the acuteeye irritation datarequirement. TOXICITYCATEGORY IIIMildly irritating to the eyes ofrabbits.Classification: AcceptableTOXICITY CATEGORY IIIMildly irritating to the eyeClassification: AcceptableToxicity Category III4855921548559233Bacillus thuringiensissubsp.kurstakistrain VBTS-2546Biopesticides Registration Action DocumentPage 36 of 38TABLE 3. ToxicologyData Requirements forBacillus thuringiensissubsp.kurstakistrain VBTS-2546, includingVBC-60219 Biological Insecticide Slurry, VBC- 60225 Biological Insecticide (Technical Powder Active Ingredient)and Two End-Use Pesticide Products (EPs), Foray 2WG and VBC-60236B Biological Insecticide Dry Flowable (40 CFR § 158.2140)Harmonized GuidelineNumberDataRequirementResultsMRID No.Slurry TGAI EP870.2500PrimaryDermalIrritationPrimary dermal irritationtesting was conducted withthe TGAI and was foundacceptable, ToxicityCategory III. Thecombination of inertingredients in this slurry istested in the TGAI and is notexpected to pose anyadditional human healthrisks via the dermal route ofexposure. It would beinappropriate andunnecessary to conducttesting with this activeingredient to evaluate thisendpoint. Thus, EPA waivedtesting for the of the primarydermal irritation datarequirement. TOXICITYCATEGORY IIIModerately irritating to the skin ofrabbits.Classification: AcceptableTOXICITY CATEGORY IIISlightly irritating to the skinClassification: AcceptableToxicity Category IV4855921648559234870.2600SkinSensitizationNANot a dermal sensitizer.Classification: AcceptableNot a dermal sensitizer.Classification: Acceptable4855921748559235Tiers II and IIINot required for Bacillus thuringiensis subsp.kurstaki strain VBTS-2546, based on the lack of acute toxicity in the Tier I studies.1According to 40 CFR § 158.2120, these data are only required for the technical grade of the active ingredient.Table 4. Non-target Organism Toxicity Requirements forBacillus thuringiensissubsp.kurstakistrain VBTS-2546(40 CFR 158.2060Requirement (OPPTSGuideline No.)MRID Toxicity and ClassificationAvian oral toxicity(885.4050)48559218The oral LD50for Northern bobwhite exposed toBtkVBTS-2546 is >1.83 x 109cfu/kg body weight per day.Classification: Acceptable.Avian inhalationtoxicity/pathogenicity(885.4100)48559236Scientific rationale was sufficientto satisfy the data requirement.BtkVBTS-2546is not considered toxic or pathogenicto birds.Classification: Acceptable.Wild mammaltoxicity/pathogenicity(885.4150)48559237 Tests required by 40 CFR § 158.2140 are adequate and appropriate for assessment of hazards to wild mammals.BtkVBTS-2546showed no adverse toxicity or pathogenicity in rats exposed via pulmonary and IV route (MRIDs 485592-10 and -11), or in toxicitytesting (MRIDs 485592-12 through -13).Classification: AcceptableFreshwater fish toxicity/pathogenicity (885.4200)48559219 Bridging rationale was submitted and two acceptable fish studies withBtkABTS-351 (MRIDs 41443405 and 41443406) were citedthat showed no treatment related effects.Classification: AcceptableFreshwater invertebratetesting(885.4240)48559220 A 21-day study shows that the EC50toDaphnia magnabased on mortality is 21 mg a.i./L (5.7 x106cfu/mL) and EC50(reproduction) was 24 mg a.i./L (6.5 x106cfu/mL).Classification: AcceptableEstuarine/Marine fishtestingEstuarine/Marineinvertebrate testing(885.4280)48559238 Bridging rationale was submitted and two acceptable marine/estuarine invertebrate studies withBtkABTS-351 (MRIDs 41540802and 41443408) were cited. These studies showed no treatment related effects to grass shrimp (Palaemonetes vulgaris) and copepod(Amphiascus minutus).Classification: AcceptableNontarget Plant Testing(885.4300)48559239 Rationale submitted was sufficient to determine that adverse effects are not expected in plants as a result of exposure toBtkVBTS2546.Classification: AcceptableNontarget Insect Testing(885.4340)48559222The 48-hour LC50was >3.9 kh/ha for an aphid parasitoid (Aphidius rhopalosophi) exposed toBtkABTS-351. The study was of tooshort duration, and themaximum hazard dose could not be confirmed.Classification: Supplemental48559223The 14-day LC50was >3.9 kh/ha for a predatory mite (Typhlodromus pyri) exposed toBtkABTS-351. The maximum hazard dosecould not be confirmed.Classification: Supplemental48559221Bridging rationale and a paper from scientific literature weresubmitted to justify no additional testing on nontarget insects.Therationale is sufficient to fulfill the data requirement.Classification: AcceptableHoneybee Testing(885.4380)48559224 The acuteoral and contactLC50was > 100 μg a.i./beefor honey bees in a 48-hour study. Thisstudy is useful for risk assessment,but is of too short duration.Classification: SupplementalAPPENDIX B. PESTICIDE PRODUCTSEPA FileSymbolRegistration NamePercentageActiveIngredientFormulation TypeUse Site(s)Method(s) ofApplicationApplicationRateTarget Pest(s)73049-UTI VBC-60219Biological InsecticideSlurry16% Manufacturing Use Product(slurry)Formanufacturinguse onlyN/A N/A N/A73049-UTO VBC- 60225Biological Insecticide(Technical PowderActive Ingredient)98% Manufacturing Use (powder) Formanufacturinguse onlyN/A N/A N/A73049-UIN VBC-60236BBiological InsecticideDry Flowable67% Dry Flowable Powder Field,Greenhouse/shadehouse,outdoor nursery,self-containedornamentalponds,ornamental treesGround and aerialapplication,chemigation1⁄4 lb /acre –1 lb/acreLooper, omnivorous leafroller, hornworm,imported cabbageworm, diamondback moth,green cloverworm, webworm, saltmarshcaterpillar, armyworm, cutworm cross-stripedcabbageworm,Heliothis virescens,Helicoverpazea, soybean looper, velvetbean caterpillar,podworm, tomato fruitworm, variegatedcutworm, melonworm, rindworm complex,fruittree leafroller, orangedog, citrus cutworm,walnut caterpillar, cankerworm, gypsy moth,variegated leafroller, redbanded leafroller,tufted apple budmoth...See label for a complete listing.73049-UIR Foray 2 WG 67% Water dispersible granules Forest & shadetrees, ornamentalfruit, nut andcitrus trees.Ground and aerialapplication1.3 – 12.8 oz.product/acreLepidopteran larvae, including gypsy moth,douglas fir tussock moth, elm spanworm, sprucebudworm, browntail moth, coneworm, buckmoth, tussock moth, pine butterfly, bagworm,leafroller, tortrix, mimosa webworm, tentcaterpillar, blackheaded budworm, saddledprominent caterpillar, saddleback caterpillar,eastern & western hemlock looper,orangestriped oakworm, satin moth, redhumpedcaterpillar, spring & fall cankerworm Californiaoakworm, fall webworm.