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, 2012
Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 

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TABLE OF CONTENTS 

I. EXECUTIVE SUMMARY ........................................................................................................................... 4

II. ACTIVE INGREDIENT OVERVIEW ....................................................................................................... 5

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

 A. Applications for Pesticide Registration .......................................................................................... 5

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

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

 A. Product Analysis Assessment (40 CFR § 158.2120) ....................................................................... 6

 B. Human Health Risk Assessment (40 CFR 158.2140) ..................................................................... 8

 C. Environmental Assessment (40 CFR § 158.2150) ........................................................................ 15

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

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

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

 A. Final Printed Labeling ................................................................................................................... 20

 B. Terms of Registration ..................................................................................................................... 20

 C. Reporting of Adverse Effects and Hypersensitivity Incidents .................................................... 20

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

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

Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 ....................................................................................... 21

 B. Environmental Protection Agency Risk Assessment Memoranda ............................................. 22

 C. Other References ............................................................................................................................ 23

APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS ........................................................... 25

APPENDIX B. PESTICIDE PRODUCTS .............................................................................................................. 38

 
Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 

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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, MPB 
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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 ingestion. 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 Valent’s DiPel® products. 

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

follows: 

VBC-60219 Biological Insecticide Slurry (73049-UTI) 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 Powder Active Ingredient (73049-UTO) is the 

technical powder, proposed for manufacturing use only, containing the 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 testing 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 Insecticide Dry Flowable and Foray 2WG) 

(EPA File Symbols 73049-UIN and 73049-UIR respectively) are effective against the listed 

economically important Lepidopteran larvae present on a large variety 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. 
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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, identification number EPA-HQ-OPP-2011-0769, and 

accessed through either http://www.regulations.gov or 

http://www.epa.gov/pesticides/regulating/registration-status.html: (1) draft Bacillus 

thuringiensis subsp. kurstaki strain VBTS-2546 Biopesticides Registration Action Document 

(BRAD); (2) draft label for the manufacturing-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 Insecticide 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, formulation types, and target pests) 

regarding the proposed pesticide products containing this active 

ingredient. 

III. REGULATORY BACKGROUND 

A. Applications for Pesticide Registration 

On August 1, 2011, Valent BioSciences Corporation, 870 Technology Way, Libertyville, IL 

60048 submitted an application for registration of four products, the manufacturing-use pesticide 

product, VBC-60219 Biological Insecticide Slurry (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 Biological Insecticide Dry Flowable (EPA File 

Symbol 73049-UIN) and Foray 2WG Biological Insecticide (EPA File Symbol 73049-UIR), 

containing the new microbial 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 products containing a new active 
Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 

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ingredient for use in forests and on shade, ornamental and sugar maple trees; on ornamental fruit, 

nut and citrus trees intended for aesthetic purposes or climatic modification that are grown in 

interior plantscapes, ornamental gardens or parks; on golf courses, lawns and grounds; and on 

agricultural crops (vegetables, fruit trees, berry bushes, nut trees, grains, herbs, alfalfa, cotton, 

flowers, bedding plants, ornamentals, hops, and greenhouse, shadehouse, outdoor nursery crops, 

and aquatic ornamentals). No comments were received following publication 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 tolerance 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 agricultural practices, provided the specifications 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 control 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 spores into each of five laboratory test mice 

weighing 17 grams to 23 grams. Such test shall 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 satisfied either by determining that each 

master seed lot brought into production is a Bacillus thuringiensis strain which does not 

produce β-exotoxin under standard manufacturing conditions or by periodically 

determining that β-exotoxin synthesized during spore production is eliminated by the 

subsequent spore-harvesting procedure. 

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

IV. RISK ASSESSMENT 

A. Product Analysis Assessment (40 CFR § 158.2120) 

Microbial pesticide product analysis data requirements1

 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. 
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Product chemistry and composition data include information about the identity of the active 

ingredient, the manufacturing process, deposition of a sample of the microbial active ingredient 

in a nationally recognized culture collection, and 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 inclusions 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 microarray 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-UTI) 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 Powder Active Ingredient (73049-UTO) is the 

technical powder, proposed for manufacturing use only, containing the 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 testing 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 Insecticide Dry Flowable and Foray 2WG) 

(EPA File Symbols 73049-UIN and 73049-UIR respectively) are effective against the listed 

economically important Lepidopteran larvae present on a large variety 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 requirements required for registration of Bacillus thuringiensis subsp. 

kurstaki strain VBTS-2546 have been fulfilled.

 
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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 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): 

In lieu of testing Bacillus thuringiensis subsp. kurstaki strain VBTS-2546, the applicant provided 

the following rationale to address the data requirement for acute oral toxicity/pathogenicity 

testing: 

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

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

kurstaki strain ABTS-351. Plasmid separation on agarose 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 literature, the method of Letowski et al. (2005)2 was used to evaluate Btk 

VBTS-2546 with slight variations. For this study3

, a newer, more comprehensive version of the 

published microarray, designated "Crickmore version 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 LD50 from this study is higher than the 

limit dose. 

In an acute oral toxicity/pathogenicity study conducted with DiPel HD-1 Technical in rats5

, a 

single dose of approximately 109 CFU/kg of body weight (kg/bw) of the test microbe was 

administered to Sprague-Dawley rats. No animals died during the test 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) 
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apparent treatment related toxicity or evidence of pathogenicity. A linear regression analysis of 

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

Based on the information presented and the lack of toxicity/pathogenicity via pulmonary6

 and 

intravenous7

 administration, Valent BioSciences Corporation is requesting that EPA agree that 

no further testing via oral administration of Bacillus thuringiensis subsp. kurstaki strain VBTS2546 is necessary to support registration. Intravenous administration 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 and acute oral toxicity studies show 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/pathogenicity conducted with DiPel Technical showed 

no evidence of toxicity or pathogenicity, and it further showed that total clearance would occur 

by 47 days. For these reasons, it would be unnecessary to conduct additional studies to evaluate 

acute oral toxicity/pathogenicity. 

 Acute Pulmonary Toxicity/Pathogenicity – Rat (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 administration of approximately 1.0 x 108 colony forming units (cfu) of 

viable technical grade test substance. The study duration was 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 pathogenic 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 approximately 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-pathogenic via intratracheal route. This study was found acceptable. 

Acute Injection Toxicity/Pathogenicity (Intravenous) – Rat (Harmonized Guideline 885.3200;

MRID No. 48559211): Bacillus thuringiensis, subsp. kurstaki strain VBTS-2546 technical 

powder was not found to be pathogenic or toxic at a dose of 6.7 x 107 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 500 days from challenge for males and females. 

The absence of clinical signs or tissue abnormalities indicated that the 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. 
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Hypersensitivity Incidents (Harmonized Guideline 885.3400): EPA regulations require that 

hypersensitivity incidents, including immediate-type and delayed-type reactions 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, 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 Guideline 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 from 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 administered 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 behavior. 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 oral toxicity 

Acute Dermal Toxicity – Rat (Harmonized 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 from a single topical application. Five 

thousand mg of Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 technical 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 clinical 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 the 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 atmosphere, 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 appeared 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 conclusion of the 14-day observation period. This study was 

found acceptable and Bacillus thuringiensis, subsp. kurstaki strain VBTS-2546 Technical 
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Powder was classified as Toxicity Category IV for 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 irritation from a single instillation via 

the ocular route. Under the conditions 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 Powder was classified as Toxicity Category III for eye irritation. 

Primary Dermal Irritation – Rabbit (Harmonized 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 irritation 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 Powder was classified as Toxicity Category III for dermal 

irritation.

Skin Sensitization – Guinea Pig (Harmonized 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 sensitization 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 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.” FFDCA section 408(p)(4), 

authorizes the Administrator, by order, to exempt 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 substance. Between October 2009 and February 

2010, EPA issued test orders/data call-ins for the first group of 67 chemicals, which contain 58 

pesticide active ingredients and nine inert ingredients. Bacillus thuringiensis is not among the 

group of 58 pesticide active ingredients on the initial list to be screened under the EDSP. 

The Agency believes that Bacillus thuringiensis likely is a substance that 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 Section 408(p) EDSP. In the event the 
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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 procedures, the list of 67 

chemicals, future lists, 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.” Section 408(c)(2)(A)(ii) of FFDCA defines “safe” to 

mean that “there is a reasonable certainty that no harm will result from aggregate exposure to the 

pesticide chemical residue, including all anticipated dietary exposures and all other exposures for 

which there is reliable information.” This includes exposure through drinking water and in 

residential settings but does not include occupational exposure. Pursuant to section 408(c)(2)(B) 

of FFDCA, in establishing or maintaining in effect an exemption from the requirement of a 

tolerance, EPA must take into account the factors set forth in section 408(b)(2)(C) of FFDCA, 

which require EPA to give special consideration to exposure of infants and children to the 

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....” Additionally, section 408(b)(2)(D) of FFDCA requires that 

EPA consider “available information concerning the cumulative effects of [a particular 

pesticide’s] . . . residues and other substances that have a common mechanism of toxicity.” 

EPA performs a number of analyses to determine the risks from 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, 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 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 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 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). 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 

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VBC-60236 is a dry flowable insecticide containing 67% active ingredient Bacillus thuringiensis

subsp. kurstaki strain VBTS-2546. Valent BioSciences Corporation 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 enhance control. Additions of 

other commonly used herbicides, insecticides, and fungicides 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, liquid fertilizers, or metal-based 

fungicides (e.g. copper sulfate-based materials). Application methods include aerial, ground and 

chemigation. 

Foray 2WG is a water dispersible granule insecticide containing 67% active ingredient Bacillus 

thuringiensis subsp. kurstaki strain VBTS-2546. Valent BioSciences Corporation intends to 

register this new product for forestry uses, shade trees, ornamentals, sugar maple trees, and 

ornamental fruits, nuts and citrus trees. Application 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 consumption of raw agricultural commodities that 

have been treated with the end use product VBC-60236 Biological Insecticide Dry Flowable. 

EPA expects the a exposure to food residues of Bacillus thuringiensis strain VBC-60236 to be 

negligible when compared to the ordinary, everyday 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 diluted prior to application and (2) rainfall and 

irrigation will further wash residues of the pesticide from the treated 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 pathogenicity demonstrated in the submitted studies. 

Foray 2WG is not labeled for application to food crops, 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, forest 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 chemigation. 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 VBC-60236 and Foray 2WG labels, residues of 

Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 are transferred to surface or ground 

waters that are intended for eventual human consumption and subjected to sanitation (e.g., 
Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 

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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: VBC-60236 is an 

agricultural-use pesticide; it is not intended to be used in and around the home, or in schools, day 

care facilities or other such settings, and its use is not expected to result in non-occupational 

residential exposure. Foray 2WG is labeled for use in forests and on ornamentals, interiorscapes, 

golf courses, and lawns. Non-occupational residential exposure may occur though contact with 

treated plants. EPA expects the aggregate exposure to residues of Foray 2WG to be negligible 

when compared to the ordinary, everyday levels to which individuals 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 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). 

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

Section 408(b)(2)(D)(v) of FFDCA requires that, when considering whether to establish, modify, 

or revoke a tolerance exemption, EPA consider “available 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 mechanism of toxicity with other 

substances; therefore, cumulative effects with other substances are not anticipated. 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 considering the establishment of a tolerance or 

tolerance exemption for a pesticide chemical residue, EPA shall assess the available information 

about consumption patterns among infants and children, special susceptibility of infants and 

children to pesticide chemical residues, and the cumulative effects on infants and children of the 

residues and other substances with a common mechanism of toxicity. In addition, FFDCA 

section 408(b)(2)(C) provides that EPA shall apply an additional tenfold (10X) margin of 

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 either retains the default value of 10X or uses 

a different additional safety factor when reliable data available to EPA support the choice of a 
Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 

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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 Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 is used as labeled in 

accordance with good agricultural practices. The toxicity 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 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 Bacillus thuringiensis

subsp. kurstaki strain VBTS-2546 when it is used as labeled and in accordance with good 

agricultural practices as an insecticide. 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 Bacillus thuringiensis subsp. kurstaki

strain VBTS-2546 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 for oral, dermal, and inhalation routes of exposure 

to Bacillus thuringiensis subsp. kurstaki strain VBTS-2546) 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. 

4. Human Health Risk Characterization 

EPA considered human exposure to Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 in 

light of the standard for registration in FIFRA and the relevant safety factors in FFDCA. A 

determination has been made that no unreasonable adverse effects to the U.S. population in 

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 literature) submitted by the applicant to support 

requests to waive nontarget organism testing for Bacillus thuringiensis subsp. kurstaki strain 
Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 

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VBTS-2546 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 Bacillus 

thuringiensis subsp. kurstaki strain VBTS-2546. 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. 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 LD50 is > 1.83 x 109cfu/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 sufficient 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 adequate and appropriate for assessment of 

hazards to wild mammals. Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 showed no 

adverse toxicity or pathogenicity in rats exposed via pulmonary and IV route (MRIDs 485592-10 

and -11), or in toxicity testing (MRIDs 485592-12 through -13). 

Nontarget Insects (Harmonized Guideline 885.4340) (MRID Nos. 48559222, 48559223

 and 48559221): No studies with nontarget insects were specifically provided for Bacillus 

thuringiensis subsp. kurstaki Strain VBTS-2546; however, two studies 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 hazard dose, given the current information about 

the test material and that labels 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 acceptable 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-351 
Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 

Biopesticides Registration Action Document 

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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 insects and some other species, and may result in 

temporary reductions in susceptible insect populations; however, the risk to nontarget insect 

populations is expected to be minimal for labeled uses as long as the product does not contain βexotoxins. 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, the acute oral and contact LC50 was determined to be > 100 mg 

a.i./bee; however, the study was supplemental because 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), and 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 the data requirement for nontarget plants. The 

microorganism Bacillus thuringiensis is well characterized, is not taxonomically related to 

known plant pathogens, and pesticide 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 the risk assessment for plants because no data are 

available for bridging. Studies with nontarget plants have typically been waived for Bacillus 

thuringiensis. The scientific rationale 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 specifically cited studies with rainbow 

trout (Oncorhynchus mykiss; MRID 41443406) and bluegill (Lepomis macrochirus; MRID 

41443405), which were included the Bt RED. The LC50 was >1.5 x 1010 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 resulting from high turbidity and suspended solids. The 

study with bluegill was classified as core (acceptable) and the LC50 was >1.5 x 1010 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 fish are not expected as a result of the proposed applications. 

Freshwater Invertebrates (Harmonized 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 EC50, based on adult immobility, was 21 mg/L (5.7 x 106

 cfu/mL) and EC50 based on 

reproduction was 24 mg/L (6.5 x 106

 cfu/mL). The NOEC value, based on mortality was 10 
Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 

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mg/L (2.7 x 106

 cfu/mL). Given the percentage of the a.i. in the EPs and the highest labeled 

application rate of 1 lb/acre, a direct application 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 may be due to physical effects 

related to excessive particles in the water. Mortality in the filtered treatment prepared with the 

same amounts as the highest treatment levels was 60%, as compared to 100%. Since exposure 

resulting from the proposed applications is well below the defined toxicity endpoints, adverse 

effects to freshwater invertebrates are not expected. This study 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 that the ABTS-351 strain is not toxic to 

marine/estuarine invertebrates, and both of these studies were classified as core (acceptable). 

Specifically, the NOEL was >2.9 x 109 cfu/g of food for grass shrimp (Palaemonetes vulgaris, 

MRID 41540802) and was 500 mg/kg (1 x 107 cfu/g sediment) for the benthic copepod 

(Amphiascus minutus, MRID 41443408). Although no fish studies were specifically cited, a 

study with the ABTS-351 strain on sheepshead minnow (Cyprinodon variegates; MRID 

41540801) is available for use in this risk assessment. In this study, the LC50 was >2.9 x 1010

cfu/L in water and >2.9 x 109 cfu/g food over a 30-day exposure period. Exposure in marine 

environments is expected to be lower than these levels (maximum hazard dosing is at least 106

cfu/mL); therefore, the proposed applications of Bacillus thuringiensis subsp. kurstaki strain 

VBTS-2546 are not expected to result in adverse effects to marine or estuarine fish or 

invertebrates. This bridging rationale is acceptable. 

2. Environmental Fate Assessment Data 

Environmental fate and groundwater 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 discussed 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 Insecticide Dry Flowable, is proposed for 

application to many types of field crops, including fruit and nut trees, watercress (when no 

standing water is present), greenhouse/shadehouse and outdoor nursery plants, aquatic 

ornamentals (self-contained, closed-system ornamental ponds only), and ornamental trees. 

Proposed application methods include ground and aerial sprays, chemigation, and direct 

application to water (for aquatic ornamentals only). The EP, Foray 2WG Biological Insecticide, 

is proposed for use on forests, shade trees, ornamentals, 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 result in contact of the a.i. with foliar surfaces and soil, and spray 

drift is expected to occur, exposure to terrestrial and aquatic nontarget organisms is possible. 

However, based on the lack of toxicity shown in studies and rationale discussed above, adverse 
Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 

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

4. Endangered Species Assessment 

EPA has determined that no effects are anticipated for most nontarget species exposed to 

Bacillus thuringiensis subsp. kurstaki, Strain VBTS-2546 as a result of the proposed labeled 

applications, including mammals, birds, terrestrial and aquatic plants, and freshwater and 

marine/estuarine animals. Although sustained population-level effects to nontarget insects are 

not expected, it is known that Bt subsp. kurstaki can affect certain species of insects (e.g., 

lepidopterans). Therefore, a “No Effect” determination is made for direct and indirect effects to 

federally listed endangered and threatened (“listed”) mammals, birds, terrestrial and aquatic 

plants, and freshwater and marine/estuarine animals and for their designated critical habitat. 

Direct effects to individuals of listed insect species cannot be precluded, nor can indirect effects 

to listed species that obligatorily depend on specific listed insects. Additional analysis of the 

potential effects to the listed taxa is necessary to make a determination for these species. 

V. ENVIRONMENTAL JUSTICE 

EPA seeks to achieve environmental justice—the fair treatment and meaningful involvement of 

all people regardless of race, color, national origin, or income—with respect to the development, 

implementation, and enforcement of environmental laws, regulations, and policies. Fair 

treatment means that no group of people, including racial, ethnic, or 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 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 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 Bacillus thuringiensis subsp. kurstaki strain 

VBTS-2546, 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 
Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 

Biopesticides Registration Action Document 

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for it; (2) its labeling and other materials required to be submitted comply with the requirements 

of FIFRA; (3) it will perform its intended function without unreasonable adverse effects on the 

environment; and (4) when used in accordance with widespread and commonly recognized 

practice, it will not generally cause unreasonable adverse effects on the environment. 

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

contradict the claims made on the labels for the manufacturing-use pesticide product, VBC60219 Biological Insecticide Slurry ,VBC-60225 Biological Insecticide Technical Powder 

Active Ingredient and/or two end-use pesticide products, VBC-60236 Biological Insecticide Dry 

Flowable and Foray 2WG Biological Insecticide, 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 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 environment, and the two EPs, VBC-60236 Biological 

Insecticide Dry Flowable and Foray 2WG Biological Insecticide in particular, are likely to 

control the caterpillar pests described on the product labels, satisfying criterion 3. Criterion 4 is 

satisfied in that Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 pesticide products are 

not expected to cause unreasonable adverse effects when used according to label instructions. 

Therefore, VBC-60219 Biological Insecticide Slurry ,VBC-60225 Biological Insecticide 

Technical Powder Active Ingredient and the two end-use pesticide products, VBC-60236 

Biological Insecticide Dry Flowable and Foray 2WG Biological Insecticide, containing Bacillus 

thuringiensis subsp. kurstaki strain VBTS-2546 as a new active 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 pesticide products containing Bacillus thuringiensis subsp. kurstaki strain 

VBTS-2546 for shipment, the registrant is required to provide appropriate 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 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. 
Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 

Biopesticides Registration Action Document 

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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 

Bacillus thuringiensis subsp. kurstaki strain VBTS-2546. 

MRID Citation 

41443401 Carter, N. (1993) Acute oral toxicity and infectivity/pathogenicity to rats of ABG 

6345, Report no. G-7239.222 

48655601 Bujor, D. (2011) VBC-60219: Product Analysis and Physical and Chemical 

Properties: Waiver Request. Project 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 Number: 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. Unpublished study prepared by Valent BioSciences 

Corporation. 4p. 

48655604 Bujor, D. (2011) VBC-60219: Acute Dermal Toxicity: Waiver Request. Project 

Number: VBC/60219/W/3. Unpublished study prepared by Valent BioSciences 

Corporation. 4p. 

48655605 Bujor, D. (2011) VBC-60219: Acute Inhalation Toxicity: Waiver Request. Project 

Number: VBC/60219/W/4. Unpublished study prepared by Valent BioSciences 

Corporation. 4p. 

48655606 Bujor, D. (2011) VBC-60219: Acute Eye Irritation: Waiver Request. Project 

Number: VBC/60219/W/5. Unpublished study prepared by Valent BioSciences 

Corporation. 4p. 

48655607 Bujor, D. (2011) VBC-60219: Primary Dermal Irritation: Waiver Request. Project 

Number: VBC/60219/W/6. Unpublished study prepared by Valent BioSciences 

Corporation. 4p. 

48677401 Bujor, D. (2011) VBC-60219: Hypersensitivity Statement. Project Number: 

VBC/60219/H1. Unpublished study prepared by Valent BioSciences Corporation. 

4p. 

48677402 Bujor, D. (2011) VBC-60225: Product Analysis and Physical and Chemical 

Properties: Waiver Request. Project 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 at 
Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 

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30(Degrees Celsius) for 18 Weeks. Project Number: 2152W. Unpublished study 

prepared by PTRL West, Inc., Abbott Laboratories and Accugenix. 87p. 

48677404 Bujor, D. (2011) VBC-60225: Hypersensitivity Statement. Project Number: 

VBC/60225/H1. Unpublished study prepared by Valent BioSciences Corporation. 

4p. 

48677405 Bujor, D. (2011) VBC-60236: Hypersensitivity 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 Pathogenicity 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 BioSciences Corporation. 6p. 

48559220 Gallagher, S.; Krueger, H. (2011) Bacillus thuringiensis, subsp. kurstaki, (Btk), 

Strain VBTS-2546: A 21-Day Life Cycle Toxicity and Pathogenicity Test with 

the Cladoceran (Daphnia magna): Final 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: VBTS/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 rhopalosiphi De Stefani Perez (Hymenoptera, 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 study prepared by Eurofins - GAB 

GmbH. 28p. 

48559236 Bujor, D. (2011) VBC-60225: Avian Inhalation Toxicity/Pathogenicity: 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 BioSciences Corporation. 5p. 

48559238 Bujor, D. (2011) VBC-60225: Estuarine/Marine Fish and Invertebrate Testing: 

Waiver Request. Project Number: VBTS/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. Unpublished study prepared by Valent BioSciences 

Corporation. 4p. 

B. Environmental Protection Agency Risk Assessment Memoranda

U.S. EPA. 2012a. Environmental risk assessment for the FIFRA Section 3 registration of 
Bacillus 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, manufacturing process and acute 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 Decision (RED): Bacillus thuringiensis. USEPA 

Office of Prevention, Pesticides, and Toxic 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 Intervals (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 No. VBTS-2546-2011-1. 

Gingras, B.A., (2011), Toxicity/ Pathogenicity Testing of VBC-60225 following Acute 

Intratracheal Challenge in rats, Report No. 2297 SN2. 

Gingras, B.A., (2011), Toxicity/ Pathogenicity Testing of VBC-60225 following Acute 

Intravenous Challenge in rats, Report No. 2297 SN1. 

Green, M., M. Heumann, R. Sokolow, L.R. Foster, R. Bryant, M.Skeels, Public Health 

Implication of the Microbial Pesticide Bacillus thuringiensis: An Epidemiological Study, 

Oregon, 1985-89, American Journal of Public 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 Polymorphism (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 nontarget invertebrates of Lepidopteran 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

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APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS 

(40 CFR PART 158 – SUBPART V) 

TABLE 1. Product Analysis Data Requirements for Bacillus thuringiensis subsp. kurstaki strain VBTS-2546, inclu

VBC-60219 Biological Insecticide Slurry, VBC- 60225 Biological Insecticide (Technical Powder Active Ingredien

and Two End-Use Pesticide Products (EPs), Foray 2WG and VBC-60236B Biological Insecticide Dry Flowable (40 CFR 

Harmonized 

Guideline 

Number 

Data Requirement 

Results

Slurry TGAI EPs 

885.1100 Product Identity 

Submitted data fulfill the requirement for product identity. VBC-60219 Biological Insecticide Slurry contains 16% by weight Bacillus 

thuringiensis subsp. kurstaki strain VBTS-2546 fermentation solids, spores, and insecticidal toxins. 

Submitted data fulfill the requirement for product identity. VBC-60225 Biological Insecticide Technical Powder contains 98% by weight 

Bacillus thuringiensissubsp. kurstaki strain VBTS-2546 fermentation solids, spores, and insecticidal toxins

Submitted data fulfill the for product identity. The 2WG and VBC-60236B 

Bacillus thuringienskurstaki strain VBTSfermentation solids, insecticidal toxins

885.1200 Manufacturing Process 

Submitted data fulfill the requirement for manufacturing process. 

885.1300 

Discussion of Formation 

of Unintentional 

Ingredients 

Submitted data fulfill the requirement for discussion of formation of unintentional ingred

N/A 

Deposition of a Sample 

in a Nationally 

Recognized Culture 

Collection 

A sample of Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 is on deposit with the Am

Culture Collection under Accession Number ATCC SD-6283 

885.1400 Analysis of Samples Rationale submitted in support 

of a request to waive this data requirement was found acceptable. 

As a term of VBC-60225 Biological Insectide TGAI registration, EPA is requiring submission of a storage stability study within one year of registration. 

Submitted data f

requirement for a

samples

885.1500 Certification of Limits Limits are adequate/acceptable. 
Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 

Biopesticides Registration Action Document 
Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 

Biopesticides Registration Action Document 

TABLE 2. Physical and Chemical Characteristics for Bacillus thuringiensis subsp. kurstaki strain VBTS-2546, inc

VBC-60219 Biological Insecticide Slurry, VBC- 60225 Biological Insecticide (Technical Powder Active Ingre

and Two End-Use Pesticide Products (EPs), Foray 2WG and VBC-60236B Biological Insecticide Dry Flowable (40 C

Harmonized Guideline Number 

Data Requirement 

Results 

Slurry TGAI EPs 

830.63021 Color Light brown Yellow-brown Not required for EP830.63031 Physical State Slurry at 20°C Solid (fine powder) at 20°C Solid 830.63041 Odor No discernible odor No discernible odor Not required for EP 830.63131 

Stability to Normal and Elevated Temperatures, Metals, and Metal Ions 

Rationale submitted in support of a request to waive this data requirement was found acceptable. 

This TGAI was prepared to be used in toxicology and ecotoxicology studies and is not used for any further formulations at this time. As a term of VBC-60225 Biological Insecticide TGAI registration, EPA is requiring submission of a storage stability study if the product is used to formulate end use products. 

Not required for EP

830.6317 Storage Stability 

Rationale submitted in support of a request to waive this data requirement was found acceptable. 

This TGAI was prepared to be used in toxicology and ecotoxicology studies and is not used for any further formulations at this time. As a term of VBC-60225 Biological Insecticide TGAI registration, EPA is requiring submission of a storage stability study if the product is used to formulate end use products. 

As a term of the registrationrequiring submission of a sstudy within one year of reg

830.63192 Miscibility Not required 

Not required because VBC-60225 Biological Insectide TGAI is not an emulsifiable liquid form of a microbial pesticide (refer to test note #2 of 40 CFR § 158.2120(d)). 

Not required
Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 

Biopesticides Registration Action Document 

TABLE 2. Physical and Chemical Characteristics for Bacillus thuringiensis subsp. kurstaki strain VBTS-2546, inc

VBC-60219 Biological Insecticide Slurry, VBC- 60225 Biological Insecticide (Technical Powder Active Ingre

and Two End-Use Pesticide Products (EPs), Foray 2WG and VBC-60236B Biological Insecticide Dry Flowable (40 C

Harmonized Guideline Number 

Data Requirement 

Results 

Slurry TGAI EPs 

830.6320 Corrosion Characteristics 

Rationale submitted in support of a request to waive this data requirement was found acceptable. 

This TGAI was prepared to be used in toxicology and ecotoxicology studies and is not used for any further formulations at this time. As a term of VBC-60225 Biological Insectide TGAI registration, EPA is requiring submission of a corrosion characteristics study if the product is used to formulate end use products. 

As a term of VBC-60236 A2WG registrations, EPA is submission of storage stabicorrosion characteristics stuone year of registration. 

830.70001 pH The pH of a 1% w/v dispersion of VBC-60219 is 4.4. The pH of a 1% w/v dispersion of VBC-60225 is 4.6. Not required for EP 830.71003 Viscosity 

Pseudoplastic behavior; range 46-270 mPa.s@ 20°C and 50- 1700 mPa.s@ 40°C 

Not required Not required 

830.7300

Density/Relative Density/Bulk Density (Specific Gravity) 

1.04 1.39

Pour density = 0.36 g/ml Tap density = 0.42 g/ml 

 1 According to 40 CFR § 158.2120, these data are only required for the technical grade of the active ingredient.  2 According to 40 CFR § 158.2120, these data are only required for emulsifiable liquid forms of microbial pesticides.

 3 According to 40 CFR § 158.2120, these data are only required for liquid forms of microbial pesticides. 
Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 

Biopesticides Registration Action Document 

TABLE 3. Toxicology Data Requirements for Bacillus thuringiensis subsp. kurstaki strain VBTS-2546, includin

VBC-60219 Biological Insecticide Slurry, VBC- 60225 Biological Insecticide (Technical Powder Active Ingredien

and Two End-Use Pesticide Products (EPs), Foray 2WG and VBC-60236B Biological Insecticide Dry Flowable (40 CFR § 158.2

Harmonized Guideline Number 

Data Requirement 

Results 

Slurry TGAI EP 

Tier I 885.30501

Acute Oral Toxicity/ Pathogenicity 

NA 

Given that acute pulmonary toxicity pathogenicity, acute injection toxicity pathogenicity and acute oral toxicity testing were conducted with the TGAI, toxicity, pathogenicity and/or infectivity in mammals is not expected following exposure to the TGAI via the oral route. The applicant submitted scientific rationale to compare Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 to previously registered strains. 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. 

N/A 
Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 

Biopesticides Registration Action Document 

TABLE 3. Toxicology Data Requirements for Bacillus thuringiensis subsp. kurstaki strain VBTS-2546, includin

VBC-60219 Biological Insecticide Slurry, VBC- 60225 Biological Insecticide (Technical Powder Active Ingredien

and Two End-Use Pesticide Products (EPs), Foray 2WG and VBC-60236B Biological Insecticide Dry Flowable (40 CFR § 158.2

Harmonized Guideline Number 

Data Requirement 

Results 

Slurry TGAI EP 

885.31501

Acute Pulmonary Toxicity/ Pathogenicity 

NA 

VBC-60225 (Bacillus thuringiensis, subsp. kurstaki, strain VBTS-2546) was not pathogenic, infective or toxic at a dose of 1.0 x 108 colony forming units (cfu) of viable technical grade (TG) test substance per animal, when administered by the intratracheal route. Classification: Acceptable N/A 

885.32001

Acute Injection Toxicity/ Pathogenicity (Intravenous) 

NA 

VBC-60225 (Bacillus thuringiensis, subsp. kurstaki, strain VBTS-2546) was not pathogenic, infective or toxic at a dose of 6.7 x 107 cfu when administered by the intravenous route. Classification: Acceptable 

N/A 

885.3400 Hypersensitiv ity Incidents 

Hypersensitivity incidents, including immediate-type and delayed-type reactions of humans or domestic animals, that occur during the testing or production of the slurry, 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)). 

885.3500 Cell Culture Not required because Bacillus thuringiensis subsp. kurstaki strain VBTS-2546is not a virus (refer to test note #4 of 40 CFR § 158.2140(d)). 
Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 

Biopesticides Registration Action Document 

TABLE 3. Toxicology Data Requirements for Bacillus thuringiensis subsp. kurstaki strain VBTS-2546, includin

VBC-60219 Biological Insecticide Slurry, VBC- 60225 Biological Insecticide (Technical Powder Active Ingredien

and Two End-Use Pesticide Products (EPs), Foray 2WG and VBC-60236B Biological Insecticide Dry Flowable (40 CFR § 158.2

Harmonized Guideline Number 

Data Requirement 

Results 

Slurry TGAI EP 

870.1100 Acute Oral Toxicity 

Acute oral toxicity testing was conducted with the TGAI and was found acceptable, Toxicity Category IV. The combination of inert ingredients in this slurry is tested in the TGAI and is not expected to pose any significant human health risks via the oral route of exposure. It would be inappropriate and unnecessary to conduct testing with this active ingredient to evaluate this endpoint. Thus, EPA waived testing for the of the acute oral toxicity data requirement. TOXICITY CATEGORY IV 

Oral LD50 female rats > 5,000 mg/kg bodyweight (bw) Classification: Acceptable TOXICITY CATEGORY IV 

 Oral LD50 female rats >5000 mg/kg bw Classification: Acceptable Toxicity Category IV 
Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 

Biopesticides Registration Action Document 

TABLE 3. Toxicology Data Requirements for Bacillus thuringiensis subsp. kurstaki strain VBTS-2546, includin

VBC-60219 Biological Insecticide Slurry, VBC- 60225 Biological Insecticide (Technical Powder Active Ingredien

and Two End-Use Pesticide Products (EPs), Foray 2WG and VBC-60236B Biological Insecticide Dry Flowable (40 CFR § 158.2

Harmonized Guideline Number 

Data Requirement 

Results 

Slurry TGAI EP 

870.1200 Acute Dermal Toxicity 

Acute dermal toxicity testing was conducted with the TGAI and was found acceptable, Toxicity Category IV. The combination of inert ingredients in this slurry is tested in the TGAI and is not expected to pose any significant human health risks via the dermal route of exposure. It would be inappropriate and unnecessary to conduct testing with this active ingredient to evaluate this endpoint. Thus, EPA waived testing for the of the acute dermal toxicity data requirement. TOXICITY CATEGORY IV 

Dermal LD50 combined (male and female) rats > 5,000 mg/kg Classification: Acceptable TOXICITY CATEGORY IV 

Dermal LD50 combined (male and female) rats > 5,000 mg/kg Classification: Acceptable Toxicity Category IV 
Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 

Biopesticides Registration Action Document 

TABLE 3. Toxicology Data Requirements for Bacillus thuringiensis subsp. kurstaki strain VBTS-2546, includin

VBC-60219 Biological Insecticide Slurry, VBC- 60225 Biological Insecticide (Technical Powder Active Ingredien

and Two End-Use Pesticide Products (EPs), Foray 2WG and VBC-60236B Biological Insecticide Dry Flowable (40 CFR § 158.2

Harmonized Guideline Number 

Data Requirement 

Results 

Slurry TGAI EP 

870.1300 

Acute Inhalation Toxicity 

Acute inhalation toxicity testing was conducted with the TGAI and was found acceptable, Toxicity Category IV. The combination of inert ingredients in this slurry is tested in the TGAI and is not expected to pose any significant human health risks via the inhalation route of exposure. It would be inappropriate and unnecessary to conduct testing with this active ingredient to evaluate this endpoint. Thus, EPA waived testing for the of the acute inhalation toxicity data requirement. TOXICITY CATEGORY IV 

Inhalation LC50 combined (male and female) rats > 5.16 mg/L Classification: Acceptable TOXICITY CATEGORY IV 

LC50 combined (male and female) rats >5.24 mg/L Classification: Acceptable Toxicity Category IV 
Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 

Biopesticides Registration Action Document 

TABLE 3. Toxicology Data Requirements for Bacillus thuringiensis subsp. kurstaki strain VBTS-2546, includin

VBC-60219 Biological Insecticide Slurry, VBC- 60225 Biological Insecticide (Technical Powder Active Ingredien

and Two End-Use Pesticide Products (EPs), Foray 2WG and VBC-60236B Biological Insecticide Dry Flowable (40 CFR § 158.2

Harmonized Guideline Number 

Data Requirement 

Results 

Slurry TGAI EP 

870.2400 Acute Eye Irritation 

Acute eye irritation testing was conducted with the TGAI and was found acceptable, Toxicity Category III. The combination of inert ingredients in this slurry is tested in the TGAI and is not expected to pose any additional human health risks via the eye route of exposure. It would be inappropriate and unnecessary to conduct testing with this active ingredient to evaluate this endpoint. Thus, EPA waived testing for the of the acute eye irritation data requirement. TOXICITY CATEGORY III 

Mildly irritating to the eyes of rabbits. Classification: Acceptable TOXICITY CATEGORY III 

Mildly irritating to the eye Classification: Acceptable Toxicity Category III 
Bacillus thuringiensis subsp. kurstaki strain VBTS-2546 

Biopesticides Registration Action Document 

TABLE 3. Toxicology Data Requirements for Bacillus thuringiensis subsp. kurstaki strain VBTS-2546, includin

VBC-60219 Biological Insecticide Slurry, VBC- 60225 Biological Insecticide (Technical Powder Active Ingredien

and Two End-Use Pesticide Products (EPs), Foray 2WG and VBC-60236B Biological Insecticide Dry Flowable (40 CFR § 158.2

Harmonized Guideline Number 

Data Requirement 

Results 

Slurry TGAI EP 

870.2500 

Primary Dermal Irritation 

Primary dermal irritation testing was conducted with the TGAI and was found acceptable, Toxicity Category III. The combination of inert ingredients in this slurry is tested in the TGAI and is not expected to pose any additional human health risks via the dermal route of exposure. It would be inappropriate and unnecessary to conduct testing with this active ingredient to evaluate this endpoint. Thus, EPA waived testing for the of the primary dermal irritation data requirement. TOXICITY CATEGORY III 

Moderately irritating to the skin of rabbits. Classification: Acceptable TOXICITY CATEGORY III 

Slightly irritating to the skin Classification: Acceptable Toxicity Category IV 

870.2600 Skin Sensitization NA Not a dermal sensitizer. Classification: Acceptable Not a dermal sensitizer. Classification: Acceptable Tiers II and III Not required for Bacillus thuringiensis subsp. kurstaki strain VBTS-2546, based on the lack of acute toxicity in the Tier I studies. 

 1 According 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 for Bacillus thuringiensis subsp. kurstaki strain VBTS-2546(40 CFR 158.2060 

Requirement (OPPTS Guideline No.) 

MRID Toxicity and Classification 

Avian oral toxicity  (885.4050) 

48559218 The oral LD50 for Northern bobwhite exposed to Btk VBTS-2546 is > 1.83 x 109 cfu/kg 

Classification: Acceptable. 

Avian inhalation toxicity/ pathogenicity (885.4100) 

48559236 Scientific rationale was sufficient to satisfy the data requirement. Btk VBTS-2546 is not 

to birds. Classification: Acceptable. 

Wild mammal toxicity/ pathogenicity (885.4150) 

48559237 Tests required by 40 CFR § 158.2140 are adequate and appropriate for assessment of hazards to 

showed no adverse toxicity or pathogenicity in rats exposed via pulmonary and IV route (MRIDs testing (MRIDs 485592-12 through -13). Classification: Acceptable 

Freshwater fish toxicity/ pathogenicity (885.4200) 

48559219 Bridging rationale was submitted and two acceptable fish studies with Btk ABTS-351 (MRIDs 4

that showed no treatment related effects. Classification: Acceptable

Freshwater invertebrate testing  (885.4240) 

48559220 A 21-day study shows that the EC50 to Daphnia magna based on mortality is 21 mg a.i./L (5.7 x1

(reproduction) was 24 mg a.i./L (6.5 x106 cfu/mL). 

Classification: Acceptable

Estuarine/Marine fish testing Estuarine/Marine invertebrate testing (885.4280) 

48559238 Bridging rationale was submitted and two acceptable marine/estuarine invertebrate studies with B

and 41443408) were cited. These studies showed no treatment related effects to grass shrimp (Pa

(Amphiascus minutus). 

Classification: Acceptable

Nontarget Plant Testing (885.4300) 

48559239 Rationale submitted was sufficient to determine that adverse effects are not expected in plants as 

2546. Classification: Acceptable

Nontarget Insect Testing (885.4340) 

48559222 The 48-hour LC50 was >3.9 kh/ha for an aphid parasitoid (Aphidius rhopalosophi) exposed to Btk 

short duration, and the maximum hazard dose could not be confirmed. 

Classification: Supplemental

48559223 The 14-day LC50 was >3.9 kh/ha for a predatory mite (Typhlodromus pyri) exposed to Btk ABTS

could not be confirmed. Classification: Supplemental

48559221 Bridging rationale and a paper from scientific literature were submitted to justify no additional te

rationale is sufficient to fulfill the data requirement. Classification: Acceptable

Honeybee Testing (885.4380) 

48559224 The acute oral and contact LC50 was > 100 μg a.i./bee for honey bees in a 48-hour study. This stu

but is of too short duration. Classification: Supplemental
APPENDIX B. PESTICIDE PRODUCTS 

EPA File Symbol 

Registration Name Percentage 

Active Ingredient 

Formulation Type Use Site(s) Method(s) of 

Application 

Application Rate 

Target P

73049-UTI VBC-60219 

Biological Insecticide Slurry 

16% Manufacturing Use Product 

(slurry) 

For manufacturing use only 

N/A N/A N/A 

73049-UTO VBC- 60225 

Biological Insecticide (Technical Powder Active Ingredient) 

98% Manufacturing Use (powder) For 

manufacturing use only 

N/A N/A N/A 

73049-UIN VBC-60236B 

Biological Insecticide Dry Flowable 

67% Dry Flowable Powder Field, 

Greenhouse/ shadehouse, outdoor nursery, self-contained ornamental ponds, ornamental trees 

Ground and aerial application, chemigation 

1⁄4 lb /acre – 1 lb/acre 

Looper, imported green clocaterpillacabbagewzea, soybpodwormcutwormfruittree walnut cvariegatetufted apSee label 

73049-UIR Foray 2 WG 67% Water dispersible granules Forest & shade 

trees, ornamental fruit, nut and citrus trees. 

Ground and aerial application 

1.3 – 12.8 oz. product/acre 

Lepidoptdouglas budwormmoth, tusleafrollercaterpillapromineneastern & orangestrcaterpillaoakworm