&EPA 

Bacillus sphaericus 

EPA-HQ-OPP-2013-0116 

Case 6052 

Preliminary Work Plan and Summary Document 

Registration Review: Initial Docket 

March 2013 

Bacillus sphaericus strain 2362serotype H5a5b strain ABTS 17 43 

(PC codes 119801, 119803) 

Bacillus sphaericus 2362 serotype H5a5b strain AM614 

(PC Code 119801, 119804) 

Approved by: 

:ft{z~~ Keith A. Matthews, Director 

Biopesticides and 

Pollution Prevention Division 

Date:~ ~?~ ~/~ 
Bacillus splwericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations.gov 

2 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations. gov 

TABLE OF CONTENTS 

A. Overview ...................................................................................................................... 5 

B. Statutory and Regulatory Authority .............................................................................. 6 

C. Background and Regulatory Information ..................................................................... 7 

D. Anticipated Data Needs and Risk Assessments ........................................................ 10 

E. Risk to Threatened and Endangered Species ........................................................... 19 

F. Product Performance ................................................................................................. 19 

G. Endocrine Effects ....................................................................................................... 19 

H. Incidents .................................................................................................................... 20 

I. Timeline ..................................................................................................................... 20 

J. Guidance for Commenters ......................................................................................... 21 

K. Next Steps ................................................................................................................. 22 

II. FACT SUMMARY FOR CASE 6052 .................................................................................. 23 

Ill. BIBLIOGRAPHY ......................................................................................... ~ ...................... 24 

A. Studies Supporting Registration Review of Bacillus sphaericus strain ABTS 17 43 (PC 

Codes 119801 , 119803) ............................................................................................ 24 

B. Studies Supporting Registration Review of Bacillus sphaericus serotype H5a5b strain 

AM614 (PC code 119801, 119804) ........................................................................... 28 

C. Environmental Protection Agency Risk Assessment Memoranda .............................. 29 

3 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations. gov 

ACRONYMS AND ABBREVIATIONS 

4SstarM FourStar Microbials, LLC (Company Number 83362) 

4SstarMP FourStar Microbial Products, LLC (Company Number 85685) 

AM Advanced Microbiologicals, LLC. (Company Number 84268) 

B. sphaericus Bacillus sphaericus 

BSIU Bacillus sphaericus International Units 

ABTS 1743 Bacillus sphaericus strain 2362 serotype H5a5b ABTS 1743 

AM614 Bacillus sphaericus strain 2362 serotype H5a5b AM614 (also known as SPH88) 

BPPD EPA's Biopesticides and Pollution Prevention Division 

C.F.R. Code of Federal Regulations 

cfu Colony forming units 

DER Data Evaluation Record- refers to EPA's review of company submissions of data or information 

DNA Deoxyribonucleic acid 

EDSP Endocrine Disrupter Screening Program 

EP End-use pesticide product 

EPA U.S. Environmental Protection Agency (the Agency) 

FFDCA Federal Food, Drug, and Cosmetic Act 

FIFRA Federal Insecticide, Fungicide, and Rodenticide Act 

FQPA Food Quality Protection Act 

FR Federal Register 

FWP Final Work Plan 

gal Gallon 

g gram 

HR Hypersensitive response 

IDS Incident Data System 

kD Kilodalton 

kg Kilogram 

lb Pound 

memo memorandum 

mg Milligram 

MRID No. Master Record Identification Number: EPA's system of recording and tracking studies submitted 

to the Office of Pesticide Programs. 

MP Manufacturing-use Product 

MRL Maximum Residue Limit 

OPP EPA's Office of Pesticide Programs 

oz Ounce 

PC Code Pesticide chemical code: a six-digit number assigned by OPP to identify pesticide chemicals. 

PWP Preliminary Work Plan 

TGAI Technical grade of the active ingredient 

TMDL Total Maximum Daily Load 

u.s. United States 

VBS Valent BioSciences (Company Number 73049) 

4 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations. gov 

I. PRELIMINARY WORK PLAN 

A. Overview 

The docket for Bacillus sphaericus (Case 6052) is now open, initiating the first public comment 

period for this registration review (docket number EPA-HQ-OPP-2013-0116). This case 

includes the following active ingredients: Bacillus sphaericus 2362 serotype H5a5b ABTS 17 43 

(PC codes 119801, 119803) and Bacillus sphaericus 2362 serotype H5a5b strain AM614 (also 

known as SPH88) (PC Codes 119801, 119804). Bacillus sphaericus are aerobic endosporeforming bacteria belonging to the family Bacilliaceae, order Bacil/iales, class Bacilli, phylum 

Firmicutes, and kingdom Eubacteria. Entomopathogenic strains of B. sphaericus can be found 

worldwide in water, soil, and insects. Bacterial insecticide pesticide products containing these 

active ingredients are registered under the Federal Insecticide, Fungicide and Rodenticide Act 

(FIFRA) for use against selected species of mosquito larvae in aquatic habitats, including crop 

sites, where mosquitoes grow. 

These two active ingredients will be referred to as ABTS 1743 or AM614 in this document 

pending confirmation of the consistency in nomenclature and labeling during this registration 

review. Bacillus sphaericus strains are included in registered Manufacturing-Use and End-Use 

products (MPs and EPs, respectively). The current registrants are as follows: 

• Valent BioSciences (called VBS in this document)- MP and EPs- company number 

73049; 

• Advanced Microbiologicals, LLC (called AM in this document)- MP and EPs- company 

number 84268; 

• B2E doing business as (dba) FourStar Microbials, LLC- EPs- company number 83362; 

and 

• FourStar Microbial Products, LLC - EPs - company number 85685). 

This Preliminary Work Plan (PWP) document explains what the Environmental Protection 

Agency's (EPA) Office of Pesticide Programs knows about the registered pesticides containing 

the Bacillus sphaericus strains in this registration review case, highlighting anticipated data 

and assessment needs, identifying types of information that would be especially useful to EPA 

in conducting the review, and providing an anticipated timeline for completing the review of this 

Case 6052. 

The Registration Review process was designed to include a public participation component to 

solicit input from interested stakeholders. EPA intends, by sharing this information in the 

docket, to inform the public of what it knows about the registered Bacillus sphaericus strains 

and what types of new data or other information would be helpful for EPA to receive as it 

moves toward a decision on Bacillus sphaericus. EPA encourages all interested stakeholders 

to review the PWP and to provide comments and additional information that will help EPA's 

decision-making process for these microbial pesticides. In addition to general areas on which 

persons may wish to comment, there are some areas identified in the PWP about which EPA 

specifically seeks comments and information. Interested stakeholders could include the 

following: environmental nonprofit or interest groups; pesticide manufacturers; agricultural 

labor or commodity groups; commercial, institutional, residential, and other users of pesticides; 

or the public at large. 

5 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www.regulations.gov 

The PWP begins by discussing the statutory and regulatory authority for Registration Review. 

Next, it provides background on Bacillus sphaericus currently registered pesticide products, 

use sites, and tolerance exemption information. Then, it lists the anticipated data needs and 

risk assessments and a projected Registration Review timeline for Bacillus sphaericus strains 

in Case 6052. Table 1 lists the currently registered B. sphaericus pesticides. Additional 

information about application rates and methods for these registered pesticides is available in 

the labels on EPA's website http://www.epa.gov/pesticides/ppls. Finally, the PWP describes 

guidance for commenters, explains the next steps EPA will be taking for this Registration 

Review, summarizes background information for this Registration Review, and lists supporting 

studies, risk assessment memoranda, and other documents. 

Further information about this case, including background and supporting documents, is 

available at http://www.regulations.gov (under "SEARCH for: Rules, Comments, Adjudications 

or Supporting Documents," enter the docket identification number, which is indicated above). 

B. Statutory and Regulatory Authority 

The Food Quality Protection Act (FQPA) of 1996 mandated a Registration Review program. All 

pesticides distributed or sold in the United States (U.S.) must generally be registered by EPA, 

based on scientific data showing that they will not cause unreasonable risks to human health 

or the environment when used as directed on the product labeling. The Registration Review 

program is intended to make sure that, as the ability to assess risk evolves and as policies and 

practices change, all registered pesticides continue to meet the statutory standard of no 

unreasonable adverse effects to human health or the environment. Changes in science, public 

policy, and pesticide use practices will occur over time. Through the Registration Review 

program, EPA periodically reevaluates pesticides to make sure that, as change occurs, 

products in the marketplace can continue to be used safely. Information on this program is 

provided on EPA's website. 1 

EPA is implementing the Registration Review program pursuant to Federal Insecticide, 

Fungicide, and Rodenticide Act (FIFRA) section 3(g) and will review each registered pesticide 

every 15 years to determine whether it continues to meet the FIFRA standard for registration. 

The regulations governing Registration Review begin at 40 C.F.R. § 155.40. EPA will consider 

benefits information and data as required by FIFRA. The public phase of Registration Review 

begins when the initial docket is opened for each case. The docket is EPA's opportunity to 

state what it knows about the pesticide and what additional risk analyses and data or 

information it believes are needed to make a Registration Review decision. After reviewing and 

responding to comments and data received in the docket during this initial comment period, 

EPA will develop a Final Work Plan (FWP) and anticipated schedule for the Registration 

Review of the Bacillus sphaericus strains 2362 serotype H5a5b ABTS 17 43 and AM614. 

1 See http://www.epa.gov/oppsrrdl/registration review!. 

6 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations. gov 

C. Background and Regulatory Information 

i. Summary of the Active lngredients2 

Baci/lus sphaericus are aerobic endospore-forming bacteria belonging to the family 

Baci/liaceae, order Bacilliales, class Baci/li, phylum Firmicutes, and kingdom Eubacteria. Both 

insecticidal and non-insecticidal entomopathogenic strains of B. sphaericus can be found 

worldwide in water, soil, and insects. Some B. sphaericus strains may contain parasporal 

inclusions, but may not be entomopathogenic. 

The Baci/lus sphaericus strains addressed in this document are identified as strain 2362 by 

cellular fatty acid analysis and included within the H5a5b serotype group by flagellar antigen 

analysis (MRID 40885301, U.S. EPA Memorandum dated 03/10/89 from W. Hazel toP. 

Hutton). Baci/lus sphaericus strain 2632 serotype H5a5b (now called ABTS 17 43) was first 

isolated in 1981 from an adult blackfly (Simulium damnosum) in Nigeria and subsequently 

obtained in the U.S. These strains can be uniquely identified by their antibiotic resistance 

profile and from the morphology of crystalline inclusions (toxin proteins) produced during 

sporulation. While they are both characterized as Baci/lus sphaericus 2362 serotype H5a5b, 

the two Bacillus sphaericus strains addressed in this document are differentiated as strains 

ABTS 17 43 and AM614. 

AM provided a written certification from the lnstitut Pasteur that AM614 (also known as SPH88) 

is B. sphaericus serotype H5a5b, strain 2362 (Attachment 1 of MRID 47356801, U.S. EPA 

Memorandum 2008c from I. Barsoum to M. Mendelsohn). AM also provided comparisons of 

the biochemical profiles of AM614 to demonstrate its similarity to VBS' ABTS 1743 active 

ingredient. The similarities were demonstrated by using the following tests: 1) standardized 

identification system Baci/lus for microbes; 2) cellular fatty acid profile (FAME) analysis; and 3) 

Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) analysis (U.S. EPA 

Memoranda 2008c, 2011 e, 2011f, from I. Barsoum to M. Mendelsohn). 

Strain ABTS 1743 was assigned PC Code 119801 when it was first registered on August 21, 

1991 to Abbott Labs, as an EP with limited use for mosquito control (EPA Reg. No. 275-77; 

Caswell No. 143447-72-7). It was transferred on April29, 2000, to VBS and is now called 

Vectolex CG Biological Larvicide (EPA Reg. No. 73049-20). VBS registered a technical product 

containing 11% ABTS 17 43 on September 27, 2004 (EPA Reg. No. 73049-65). Other 

amendments were made to update the registrations of three EP pesticide formulations 

containing the active ingredient ABTS 1743 (see Table 1). EPA recoded ABTS 1743 as PC 

Code 119803 on March 15, 2012 to distinguish it from the other registered strain. 

On September 15, 2008, AM registered the other B. sphaericus 2362 serotype H5a5b strain 

AM614 as a manufacturing use product (EPA Reg. No. 84268-1) for formulating into EPs. At 

the time of the FIFRA section 3(c)(7) registration, AM614 was called SPH88 and was also 

assigned PC code 119801 . During the registration review process, EPA anticipates requiring 

clarification of the nomenclature and consistency in labeling of AM's B. sphaericus pesticides. 

AM demonstrated similarity to ABTS 1743 as previously discussed. On March 15, 2012, EPA 

recoded AM614 to PC Code 119804 to distinguish it from VBS' active ingredient, but data 

2 Information in this section extracted from U.S. EPA (1989a), U.S. EPA. (2008c) and labels. 

7 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations.gov 

submissions and reviews between 2008 and 2012 may refer to AM614 products as PC code 

119801. 

Products containing Bacillus sphaericus are microbial mosquito larvicides registered for use 

where mosquitoes live and lay eggs, e.g. storm water and drainage systems, marine and 

coastal areas, freshwater bodies such as lakes and streams, water that collects in discarded 

tires, manmade and natural bodies of water, dormant rice fields, and water in agricultural sites. 

Pesticides containing these registered Bacillus sphaericus strains are not registered for use in 

or on treated or finished drinking water for human consumption. Additional details of these use 

sites, application rates and types are in Table 1 below, and also can be found on the pesticide 

product labels; see: http://www.epa.gov/pesticides/ppls. 

ii. Bacillus sphaericus Products 

In 1991, EPA registered the first Bacillus sphaericus -containing pesticide product for use to 

control mosquito larvae. There are currently ten FIFRA Section 3 registered B. sphaericus 

insecticide products3

, two of these products are MPs. VBS and AM have each registered aMP 

to be used for formulating into EPs to control selected species of mosquito larvae in a variety 

of habitats, including crop sites. Some of the registered EPs also contain Bacillus thuringiensis. 

Registered pesticide products containing both these active ingredients are available as 

briquets, controlled release granular (CRG), granular, spray, wettable powder and water 

soluble pouch formulations for application both by ground and aerially. See Table 1 below for 

additional information on the currently registered Bacillus sphaericus products. 

TABLE 1. Summary of Currently Registered Bacillus sphaericus Products 

EPA Application 

Registration Product Name Rates 

Number and (% Active Ingredient, (range) Application Use Sites** 

Date Registered Fonnulation Type) IU=BS Methods 

International 

Units* 

Bacillus sphaericus strain 2362 serotype H5a5b ABTS 1743 (PC Codes 1198012, 1198032) 

73049-651 Bacillus sphaericus N/A Manufacturing use For manufacturing use only. 09/27/04 Slurry 11% 

Mosquito habitats - agricultural crop and 

non-crop sites Wastewater, Stormwater/ 

151U- 80 Drainage Systems, Marine/ Coastal Areas, 

73049-20 Vectolex CG Biological lbs/acre For ground and Water Bodies, Rice, pastures/hay fields, 

08/21/1991 Larvicide 7.5% granular aerial equipment orchards, citrus groves, irrigated crops. 

Water soluble pouch for ponds, lagoons, 

water gardens, bird baths, fountains, 

flower pots and planters. 

Vectolex WDG Wastewater, Stormwater/Drainage 

73049-57 Biological Larvicide 76.81U -0.5 For ground and Systems, Marine/Coastal Areas, Water 

06/08/1999 (51.2%) (dried 1.5 lbs/ acre aerial equipment Bodies (natural and manmade), 

concentrate) granular- Agricultural crops (Dormant Rice Fields, 

for organic use Rice, pastures/hay fields, orchards, citrus 

3 Section 3 product labels can be obtained from the Pesticide Product Label System (PPLS) website 

(http :1/oaspub. epa. gov/pestlab/Jppls .home or www. epa. gov/pesticides/ppls). 

8 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations.gov 

TABLE 1. Summary of Currently Registered Bacillus sphaericus Products 

EPA Application 

Registration Product Name Rates 

Number and (% Active Ingredient, (range) Application Use Sites** 

Date Registered IU=BS Methods Formulation Type) International 

Units* 

groves, irrigated crops. Rice fields, 

pastures/hay fields, orchards (including 

citrus groves, peaches, almonds, dates, 

and walnuts), asparagus fields, corn fields, 

cotton fields, alfalfa fields, vineyards. 

Vectomax FG For ground and 

73049-429 Biological Larvicide 541U-20 aerial equipment; Sites that contain fish, other aquatic life 

04/24/2006 2.7% B.sphaericus lbs/acre water soluble and plants +4.5% Bti 4 includes 

water soluble pouch pouch 

Bacillus sphaericus strain AM614 (PC Codes 1198013, 1198043) Advanced Microbiologies LLC (84268) MP and EP 

84268-1 1 AM61432 Primary 

09/15/2008 Powder 100% N/A Manufacturing use For manufacturing use only. 

Powder 

Waste water, storm water/drainage 

84268-2 AM61432 (50 G) 100 IU to For ground and systems, Marine coastal areas, natural and 

09/15/2008 5% granular 5-20 lb/acre. aerial equipment man-made water bodies, dormant rice 

fields. 

Rice fields, pastures I hay fields, orchards, 

citrus groves, irrigated crops, agricultural 

84268-3 AM61432 WP 97.51U For ground and irrigation waters (0.5 to 1.5 lb/acre); 

09/15/2008 65% - wettable powder 0.5 to 1.5 aerial spraying Wastewater, storm water/drainage 

lb/acre systems, Marine coastal areas, natural and 

man-made water bodies, dormant rice 

fields. 

Bacillus sphaericus strain AM614 EPs (PC Codes 1198013, 1198043) FourStar Products (83362) 

and FourStar Microbial Products (85685) 

Areas that contain aquatic life, fish and 

plants, or used by or in contact with 

humans, animals, horses, livestock, pets, 

birds or wildlife. Storm drains, catch basins, 

underground drainage systems, storm 

83362-3 FourStar Briquets Briquet Briquet - Apply by water retention areas, retention ponds, 

05/06/2009 6% hand abandoned swimming pools, ornamental 

fountains, ponds, fish ponds, water 

gardens, animal drinking troughs, standing 

water, water holding receptacles, manmade and natural sites where mosquitoes 

grow. 

FourStar CRG 180 IU- to 20 Add granule or Mosquito habitats- Pre-hatch or post85685-2 (Controlled Release lbs/acre; one pouch to hatch to temporary and permanent water 

09/07/2011 Granule) ( 1 ) teaspoon treatment area by sites prior to flooding; areas where aquatic 

+ 1% BtF - Granule or - cup/50-2400 hand; ground or life, fish, plants, humans, animals, horses, 

Water soluble pouch square foot aerial application. livestock, pets, birds or wildlife; freshwater 

9 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations.gov 

TABLE 1. Summary of Currently Registered Bacillus sphaericus Products 

EPA Application 

Registration Product Name Rates 

Number and {% Active Ingredient, {range) Application Use Sites** 

Date Reglstttred Formulation Type) IU=BS Methods 

International 

Units* 

water; 1-2 marshes, cattail marshes, woodland pools 

pouches/50 and meadows, grassy swales, floodplains, 

square ft- flood water, standing water, roadside 

double rate in ditches, canals, catch basins, storm drains, 

deep water, storm water collection areas and 

dense surface impoundments, lakes, ponds, abandoned 

cover swimming pools, ornamental fountains, 

ponds, water gardens, water holding 

receptacles, other natural and manmade 

sites. 

Commercial and Residential Use [including 

Water Soluble Pouches (WSP)] Any 

85685-3 FourStar MBG 60 IU-Ibs/acre Ground, aerial location where water accumulates and 

10/25/2012 + 3% Bti 5 to 20 lbs/acre applications remains standing for periods of time, 

except treated, finished drinking water for 

human consumption. 

Footnotes Table 1: 1Manufactunng Use Product. 28. sphaencus serotype H5a5b stra1n ABTS1743. 38. sphaencus serotype 

H5a5b strain AM614. 48. thuringiensis israelensis (Btt) H-14 strain AM65-52 5 Bacillus thuringiensis israe/ensis (Btt) Strain 

BMP144. *The potency of the active ingredient is expressed as B.sphaericus International Units (BS IU) rather than cfu/g. . 

**For details of sites see use s~es on EPA's label site at www.epa.gov/pesticides/ppls e.g .. Waste tires -1 VectoLex Soluble Pouch for up to 50 

square feet of treatment area. 

iii. Tolerance Actions 

An exemption from the requirement of a tolerance is established for residues of the microbial 

pesticides, Bacillus sphaericus when used in or on all food crops [40 CFR § 180.1202, 63 FR 

48597, Sept. 11 , 1998]. Registered Bacillus sphaericus pesticides can be applied to 

agricultural food crop areas where water is standing, such as dormant rice fields, etc. (see 

Table 1 and labels). In light of available toxicity and exposure data, EPA concluded that there 

was a reasonable certainty that no harm would result to the U.S. population, including infants 

and children, from aggregate exposure to residues of Bacillus sphaericus (Final Rule to 

establish tolerance exemption, 63 FR 48597, Sept. 11, 1998). Since this tolerance exemption 

took effect after FQPA amended FIFRA in 1996, the exemption did not require reassessment. 

D. Anticipated Data Needs and Risk Assessments 

Provided below are tables of the current data requirements for microbial pesticides as set forth 

in 40 C.F.R. part 158, subpart U. They are arrayed by scientific discipline (i.e., product 

chemistry, toxicology, and nontarget organisms and environmental fate) and descriptions are 

included regarding how the data requirements have been fulfilled or where data gaps 

(anticipated data needs) have been identified. 

Presently, there are data gaps and anticipated data needs for both registered strains of 

Bacillus sphaericus, as follows: 

10 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations. gov 

i. For technical products for both ABTS 1743 and AM614, confirmatory data/information 

that the Product Analysis requirement has been satisfied for 'deposition of a sample in a 

nationally recognized culture collection' (40 CFR 158.2120) and meets current Agency 

data requirements for registration of pesticides under FIFRA. 

i. Product Chemistry (40 C.F.R. § 158.2120) 

Tables 2 to 5 below provide summaries of product chemistry data that support registration of 

Bacillus sphaericus strain ABTS 1743 and AM614. The data continue to meet EPA's current 

data requirements for pesticide registration. 

ABTS 1743 

One EP (EPA Reg. No. 275-77) was registered to Abbott in 1991 and transferred to VBS as 

EPA Reg. No. 73049-20). VBS cited the previously submitted and reviewed EP data and 

provided additional data to register the MP (EPA Reg. No. 73049-65). All product chemistry 

requirements for registration of products containing ABTS 1743 have been met. Nevertheless, 

EPA anticipates requiring confirmation that the deposit of active ingredient in a National or 

recognized culture collection is currently maintained to validate that the source of the active 

ingredient is still the same as was originally registered. 

TABLE 2. Summary of Product Chemistry Data for Bacillus sphaericus strain ABTS 1743 Technical . 

EPA Reg. No. 73049-65 

Harmonized Has Data MRID/ Accession 

Guideline Data Requirement Requirement Been Source Number or Other 

Number Addressed? How? Reference 

U.S. EPA (1989a )1 40028001 

U.S. EPA (1989a)1 40885301 

885.1100 Product Identity Yes Studies U.S. EPA (1991) 41586701 

U.S. EPA (1991) 41604001 

U.S. EPA (2001) 45233004 

U.S. EPA (2004) 45681801 

U.S. EPA (1991) 41586701 

885.1200 Manufacturing Process Yes Study U.S. EPA (2001 )2 45233004 

U.S. EPA (2004)3 45681801 

Deposition of a Sample in a U.S. EPA (1989a)1 

885.1250 Nationally Recognized Culture No Data gap; ATCC 

Collection confirmation needed 

885.1300 Discussion of Formation of Yes Study 

U.S. EPA (1989a)1 40028001 

Unintentional Ingredients U.S. EPA (1991)1 41586703 

885.1400 Analysis of Samples Yes Study U.S. EPA (1991}_1 41586701 

U.S. EPA (2001)2 45233004 

885.1500 Certification of Limits Yes Study 

U.S. EPA (2001 )2 45233004 

CSF dated May 26, 2000 

1 40885301: U.S.EPA. 1989a. W. J. Hazel to P. Hutton, 03/10/89. 245233004: Memo: C. Ets1tty to A. Reynolds 10/15/01. 

345681801 :Memo C. Etsitty to A. Reynolds 09/22/04. 

II 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www.regulations.gov 

TABLE 3. Summary of Physical and Chemical Characteristics for Bacillus sphaerlcus strain ABTS 1743 (VBS 73049) 

EPA Reg. No. 73049-65 

MRID/ 

Harmonized Has Data Accession 

Guideline Number Data Requirement Requirement Been Source Number or 

Addressed? How? Other 

Reference 

830.6302 Color Yes Study U.S. EPA 1989a1 40885301 

830.6303 Physical State Yes Study U.S. EPA 40885301 (1989a)1 

830.6304 Odor Yes Study U.S. EPA 40885301 (1989a)1 

Stability to Normal and Elevated 

830.6313 Temperatures, Metals and Metal Yes -Study U.S. EPA (1991) 41586702 

Ions 

U.S. EPA (1989) 40886601 

40886602 

830.6317 Storage Stability Yes Study U.S. EPA (1991) 41586702 

U.S. EPA (2001)2 45233004 

U.S. EPA (2004)3 45681801 

830.6319 Miscibility Not required per OCSPP 830.1000(e)(2), the product is not an 

emulsifiable liquid 

830.6320 Corrosion Characteristics Yes Study U.S. EPA (1991) 41586702 

830.7000 pH Yes Study U.S. EPA (1989a) 40885301 

830.7100 Viscosity Not applicable to the TGAI test substance, as it is not a liquid. 

830.7300 Density/Relative Density/Bulk Yes Study U.S. EPA (1989a) 40885301 Density 

1 40885301: U.S.EPA. 1989a. W. J. Hazel to P. Hutton, 03/10/89. 245233004: Memo: C. Ets1tty to A. Reynolds 10/15/01. 

345681801 :Memo C. Etsitty to A. Reynolds 09/22/04. 

AM614 

AM614 was registered in accordance with FIFRA Section 3(c)(7) on September 15, 2008 with 

conditions of registration as follows: to confirm similarity to ABTS 1743, to provide methods for 

determination of unintentional ingredients, and to submit data/information regarding oxidationreduction, chemical incompatibility, storage stability, and corrosion characteristics of the active 

ingredient. AM provided the data/information required to meet these conditions (MAID 

47898202: DER I. Barsoum toM. Mendelsohn 02/10/11b; letter March 14,2011 from S. Reilly 

to A. Roberts, AM's consultant). 

The product characterization and physical and chemical properties data submitted to support 

the registration of products containing B. sphaericus strain AM614 meet the data requirements 

for microbial pesticide product analysis. As noted for the other active ingredient and for the 

same reason to ascertain the integrity of the master stock culture of the registered strain of this 

microbe, EPA anticipates requiring confirmation that the deposit in a nationally recognized 

culture collection is current (Guideline 885.1250). In addition, during the registration review 

12 
Bacillus sphaericus Case 6052 -Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations.gov 

process, EPA will clarify the relationship between AM614 and SPH88 and establish 

consistency in the nomenclature of the active ingredient in regulatory documents and labeling. 

TABLE 4. Summary of Product Chemistry Data for Bacillus sphaericus strain AM614 (EPA Reg. No. 84268-1) 

Has Data 

litarmonized Requirement MRID/Accession 

Guideline Data Requirement Been Source Number or 

Number Addressed? Other Reference 

How? 

U.S. EPA (2008c) 47157201 

47356801 1 

U.S. EPA (2008c) 47898200 

885.1100 47898202 Product Identity Yes Studies !J.S. EPA (2011 e) 47898404 

Registrant 

U.S. EPA (2011f) response to 

deficiencies 

U.S. EPA (2008c) 47157201 

885.1200 Manufacturing Process Yes Study U.S. EPA (2011a) 47356801 

U.S. EPA (2011 b) 47898202 

Yes letter from 

Pasteur U.S. EPA 2008c 

885.1250 Deposition of a Sample in a Nationally lnstitut in cited Data Gap; 47356801 Recognized Culture Collection study- need confirmation 

to ascertain needed1 

current status. 

Discussion of Formation of Unintentional U.S. EPA (2008c) 47157201 

885.1300 Yes Study 47356801 Ingredients U.S. EPA (2011 e) 47898404 

885.1400 Analysis of Samples Yes Study U.S. EPA (2008c) 47157201 

47356801 

885.1500 47157201 

Certification of Limits Yes Study U.S. EPA (2008c) 47356801 

CSF dated March 10, 2011 

Notes Table 4: 147356801, U.S. EPA Memorandum 2008c from I. Barsoum to M. Mendelsohn - letter in study states it is in the Pasteur 

lnstitut- EPA requires confirmation that culture deposit is maintained currently. 

13 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www.regulations.gov 

TABLE 5. Summary of Physical and Chemical Characteristics for Bacillus sphaericus strain AM614 

(EPA Reg. No. 84268-1) 

Has Data MRID/ Harmonized Requirement Accession Guideline Data Requirement Been Source Number or Number Addressed? Other Reference How? 

830.6302 Color Yes Study U.S. EPA (2008c) 47157201 

830.6303 Physical State Yes Study U.S. EPA (2008c) 47157201 

830.6304 Odor Yes Study U.S. EPA (2008c) 47157201 

830.6313 Stability to Normal and Elevated Yes Study U.S. EPA (2008c) 47157201 

Temperatures, Metals and Metal Ions U.S. EPA (2011b) 47898202 

U.S. EPA (2008c) 47157201 

830.6317 Storage Stability Yes Study U.S. EPA(2011b) 47898202 

U.S. EPA (2011 e) 47898404 

830.6319 Miscibility Not required per OCSPP 830.1 OOO(e)(2), the product is 

not an emulsifiable liquid. 

830.6320 Corrosion Characteristics Yes Study U.S. EPA (2011 b) 47898202 

U.S. EPA (2011 e) 47898404 

830.7000 pH Yes Study U.S. EPA (2008c) 47157201 

830.7100 Viscosity Yes Study U.S. EPA (2008c) N/A solid 

830.7300 Density/Relative Density/Bulk Density Yes Study U.S. EPA (2008c) 47157201 

ii. Toxicology (40 C.F.R. § 158.2140) 

These microbial active ingredients, B. sphaericus strains ABTS 17 43 and AM614, are not toxic, 

infective or pathogenic on the basis of acute toxicology tests conducted with laboratory 

mammals or on data/information submitted to show equivalence of strain AM614 to strain 

ABTS 1743 (see summary of active ingredients where equivalence was shown to registered 

strain by 1) standardized identification system Bacillus for microbes; 2) cellular fatty acid profile 

(FAME) analysis; and 3) Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDSPAGE) analysis (U.S. EPA Memoranda 2008c, 2011 e, 2011f, from I. Barsoum to M. 

Mendelsohn). During registration review, the EPA anticipates reviewing current labels and 

ensuring that precautionary language is present to mitigate exposure and risk to the potential 

moderate eye irritation and dermal sensitization effects of some of the formulations that include 

these active ingredients. 

14 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations. gov 

a) Strain ABTS 1743 

The toxicology data submitted to support the registration of the B. sphaericus serotype H5a5b 

strain ABTS 17 43 pesticide products satisfy the current microbial pesticide acute toxicology 

data requirements. The company's submissions for the data requirements for mammalian 

acute oral, dermal, and pulmonary toxicity, primary dermal irritation, primary eye irritation and 

dermal sensitization are shown in Tables 6 and 7 below. EPA has reviewed these submissions 

and rationale to support requests to waive testing and has determined that they meet current 

requirements for pesticide registration. The MP and EPs are in Toxicity Category IV for acute 

oral, dermal, pulmonary toxicity and primary eye irritation effects (U.S. EPA 1989a, U.S. EPA · 

1989b, and U.S.EPA 1996e). Primary dermal irritation and dermal sensitization studies indicate 

that products may be potential dermal sensitizers (U.S. EPA 1989a, U.S. EPA 1996e). 

TABLE 6. Summary of Toxicology Data for Bacillus sphaericus strain ABTS 1743 (EPA Reg. No. 73049-65) 

Harmonized Has Data Requirement MRID/ Accession 

Guideline Number Data Requirement Been Addressed? Source Number or Other 

How? Reference 

885.3050 Acute Oral Toxicity/ Yes Studies1 U.S. EPA (1989a) 40329802 

Pathogenicity U.S. EPA (1989b) 41103401 

885.3150 Acute Pulmonary Toxicity/ Yes Study1 U.S. EPA (1989a) 40329804 

Pathogenicity U.S. EPA (1989bJ 41103402 

Acute Injection Toxicity/ U.S. EPA (1989a) 40329805 

885.3200 Pathogenicity Yes Study U.S. EPA (1989b Intravenous 41103403 

40329809- inconclusive but any future hypersensitivity incidents must 

885.3400 Hypersensitivity lncidents2 be reported per 40 CFR 158.2140 (see also OPPTS Guideline 

885.3400). 

885.3500 Cell Culture Not Applicable. Bacillus sphaericus is not a virus (refer to test note #4 

of 40 CFR § 158.2140(d):. 

870.1100 Acute Oral Toxicity Yes Study U.S. EPA (1989a) 40329802 

U.S. EPA (1989b) 41103401 

870.1200 Acute Dermal Toxicity Yes Study U.S. EPA (1989b) 40329803 

870.1300 Acute Inhalation Toxicity Yes Study U.S. EPA (1989b) 40329804 

870.2400 Acute Eye Irritation Yes Study U.S. EPA (1989b) 40329808 

870.2500 Primary Dermal Irritation Yes Study U.S. EPA (1989a) 40329807 

U.S. EPA (1996e)3 439919013 

1U.S. EPA J. T. McClintock to P. Hutton 09/14/1989. 240329809, Inconclusive gu1nea p1g study submitted for th1s gUideline. 

au.S. EPA 1996e: from J. F. Heminway, 11/24/96- Tox category IV. 

15 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations. gov 

b) Strain AM614 

The toxicology data submitted for the registration of B. sphaericus serotype H5a5b strain AM614 

pesticide MP satisfy the current microbial pesticide toxicology data requirements for acute toxicity 

studies for pesticide registration (Table 7). 

TABLE 7. Summary of Toxicology Data for Bacillus sphserlcus strain AM614 (EPA Reg. No. 84268-1) 

Harmonized Has Data Requirement MAID/ Accession 

Guideline Number Data Requirement Been Addressed? Source Number or Other 

How? Reference 

885.3050 Acute Oral Toxicity/ Yes Study U.S. EPA (2008c) 47157203* Pathogenicity 

885.3150 Acute Pulmonary Yes Study U.S. EPA (2008c) 47157205 Toxicity/Pathogenicity 

Acute Injection Toxicity/ 

885.3200 Pathogenicity- Yes Study U.S. EPA (2008c) 47157203 

Intravenous 

885.3400 Hypersensitivity Incidents Any future hypersensitivity incidents must be reported per 40 CFR 

158.2140 (see also OPPTS Guideline 885.3400). 

885.3500 Cell Culture Not Applicable. Bacillus sphaericus is not a virus (refer to test note #4 of 

40 CFR § 158.2140(d)). 

870.1100 Acute Oral Toxicity Yes Study U.S. EPA (2008c) 47157203* 

870.1200 Acute Dermal Toxicity Yes Study U.S. EPA (2008c) 47157204 

870.1300 Acute Inhalation Toxicity Yes Study U.S. EPA (2008c) 47157205 

870.2400 Acute Eye Irritation Yes Study U.S. EPA (2008c) 47157206 

870.2500 Primary Dermal Irritation Yes Study U.S. EPA (2008c) 47157207 

870.2600 Dermal Sensitization Yes Study U.S. EPA (2008c) 47157208 . . . • 0 . . ... *While study 1n1t1ally class1f1ed as Supplemental at t1me of reg1strat1on, th1s was based on the results of s1m1lanty stud1es. The s1mllanty stud1es 

were submitted and found acceptable (47898202, and US. EPA 2008c, 2011e, 2011f; also see Table 4). 

iii. Nontarget Organisms and Environmental Fate (40 C.F.R. § 158.2060) 

The nontarget organism and environmental fate data submitted for registration of pesticide products 

containing the active ingredients, B. sphaericus ABTS 1743 satisfy the current microbial pesticide 

nontarget organism and environmental fate data requirements (40 CFR part 158.2150) for the currently 

labeled sites. See Tables 8 and 9 below for citations of studies and EPA reviews of these data. 

ABTS 1743 

Between 1988 and 2001, EPA reviewed data/information submitted by VBS to establish that the active 

ingredient, ABTS 1743, was not toxic or pathogenic to birds, fish, freshwater and marine invertebrates, 

nontarget insects and honeybees. The original ecological effects review (written by Z. Vaituzis, dated 

August 4, 1988) for Bacillus sphaericus H5a5b strain 2362 ABTS 17 43 provides an evaluation of the 

avian, freshwater fish, and honeybee toxicity data, and cited several data gaps. VBS subsequently 

submitted some data to satisfy the data requirements (see Table 8, for MRIDs and U.S. EPA 

16 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations. gov 

explanations and citations, including U.S. EPA (1996a-d). EPA also determined in some cases that 

data were no longer needed. The data gaps and actions to address them were: 

1) A chronic feeding study to evaluate possible risk to aquatic avian wildlife from repeated 

ingestion of B. sphaericus granular formulations. 

• EPA determined that the chronic feeding study was no longer required because it was 

initially requested on the basis of the results of an avian injection study, which is no 

longer required, because the nature of the microbial pesticide does not indicate potential 

pathogenicity to birds (Table 8). 

2) Mouse infectivity studies requested by human health effects data reviewers 

• Acute oral, acute inhalation, and acute intravenous toxicity/pathogenicity data with 

laboratory rats were subsequently submitted, the results of which concluded no toxicity, 

pathogenicity, or infectivity of Bacillus sphaericus H5a5b strain 2362 to rats. These data 

were determined to adequately address the infectivity concerns4

. 

3) Testing of green alga Selenastrum capricornutum to fulfill the Tier I nontarget plant study 

requirements. 

4) Because the current plant data requirements are conditional and only required if the 

microbial pesticide is taxonomically related to a known plant pathogen, which is not the case 

for B. sphaericus, EPA determined that this study is no longer required (Table 8: U.S. EPA 

1996b and 2008b). 

5) Estuarine and marine aquatic animal studies, since use is expected in areas with estuarine 

environments. 

• Studies with sheepshead minnow (MRID 41931301 ), mysid shrimp (MRID 41931302), 

and Eastern oyster (MRID 41931305) were submitted and were found to be acceptable 

U.S. EPA (1996a-c) 2

• The data requirement has been met. 

TABLE 8. Summary of Nontarget Organisms Data for the Bacillus sphaerlcus strain ABTS 1743 

(EPA Reg. No. 73049-65) 

Harmonized Has Data MRID/ Accession 

Guideline Data Requirement Requirement Been Source Number or Other 

Number Addressed? How? Reference 

885.4050 Avian Oral Toxicity Yes Study U.S. EPA (1988) 403298111, 

U.S. EPA (1988) 41931304 

885.4100 Avian Inhalation The nature of the microbial pesticide does not indicate potential 

Toxicity/Pathogenicity pathogenicity to birds.1 

Wild Mammal Toxicity/ Not applicable. No testing expected to be required. Results of the Tier 1 

885.4150 Pathogenicity toxicity tests required by 40 CFR § 158.2140 (Table 6) are adequate and 

appro Jriate for assessment of hazards. 

Freshwater Fish Toxicity/ 403298131 

885.4200 Yes Studies U.S. EPA (1988) 403298141 

Pathogenicity 40193701 1 

885.4240 Freshwater Invertebrate Yes Studies U.S. EPA (1988) 403298151 

Toxicity/ Pathogenicity 40193701 1 

Estuarine And Marine 41931301 

885.4280 Invertebrate Testing Yes Studies U.S. EPA (1996a-c)2 41931302 

41931305 

Nontarget plant Yes Study and U.S. EPA (1996d)2 41931303 (algae) 

885.4300 Scientific Waiver Testing Rationale U.S. EPA (2008b) Not taxonomically 

4 Memorandum from J. McClintock (OPP/HED) to P. Hutton (OPP/RD) dated September 28, 1989b and Table 2: MAID 

45681801: Memo from C. Etsitty to A. Reynolds dated 09/22/04 for acceptable mouse infectivity testing. 

17 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations. gov 

TABLE 8. Summary of Nontarget Organisms Data for the Bacillus sphaericus strain ABTS 1743 

(EPA Reg. No. 73049-65) 

Harmonized Has Data MRID/ Accession 

Guideline Data Requirement Regulrement Been Source Number or Other 

Number Addressed? How? Reference 

related to a known 

plant pathogen. 

885.4340 Nontarget insect testing Yes Scientific U.S. EPA (1988) 40391401 1 

Waiver Rationale 41604001 

885.4380 Honey bee testing Yes Scientific U.S. EPA (1988) 403298161 

Waiver Rationale .. 1VBS c1ted own data from registered 73049-20 and submitted add1t1onal data. DER 08/28/1988 from Z. Va1tuz1s toP. Hutton and2U.S. EPA 

1996 a-d: from Horne, D. dated 08/19/96 a-d. Active ingredients are very specific for mosquitoes as shown by host range specificity and 

product performance tests for both active ingredients (U.S. EPA 1999, 2006, 2008a, 2008e, 2011 g, 2012). 

AM614 

AM submitted a request to bridge data by claiming that it is similar to ABTS 17 43. Between 

2008 and 2011, AM provided data to demonstrate similarity of AM614 to the earlier registered 

active ingredient ABTS 1743 (Memoranda I. Barsoum toM. Mendelsohn, U.S.EPA 2008c, 

02/1 0 /2011 ). This led to bridging of data for avian oral toxicity pathogenicity, freshwater fish, 

estuarine/marine fish and invertebrate non-target organisms and honey bee testing, which 

were submitted to support the original registration of the ABTS 1743 strain. AM also provided a 

study with Daphnia magna and scientific rationale to waive the data requirements for 

freshwater invertebrate and nontarget insect testing, respectively (see ecotoxicology summary 

Table 9). Data to determine effects on nontarget plants were not required on the basis that 

AM614 is not known as a plant pathogen (U.S. EPA 2008b). 

TABLE 9. Summary of Nontarget Organisms Data for Bacillus sphaericus strain AM614 

(EPA Reg. No. 84268-1) 

Harmonized Has Data Requirement MRID/ Accession 

Guideline Data Requirement Been Addressed? Source Number or Other 

Number How? Reference 

Yes. Data waiver Previously submitted 

885.4050 Avian Oral Toxicity rationale in MAID U.S. EPA data are adequate to 

471572021 (2008b) fulfill data requirement 

by bridging. 

885.4100 Avian inhalation The nature of the microbial pesticide does not indicate potential 

toxicity/pathogenicity pathogenicity to birds.1 

Wild mammal toxicity/ Not applicable .No testing expected to be required. Results of the Tier 1 

885.4150 pathogenicity toxicity tests required by 40 CFR § 158.2140 (Table 7) are adequate and 

appropriate for assessment of hazards. 

Yes. Data waiver 47157202. Previously 

885.4200 Freshwater fish toxicity/ rationale in MAID U.S. EPA (2008b) submitted data are 

pathogenicity 471572021 adequate to fulfill data 

requirement by bridging. 

885.4240 Freshwater invertebrate Yes Study U.S. EPA (2008b) 47157209 toxicity/ pathogenicity 

18 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations.gov 

TABLE 9. Summary of Nontarget Organisms Data for Bacillus sphaericus strain AM614 

(EPA Reg. No. 84268-1) 

Harmonized Has Data Requirement MRID/ Accession 

Guideline Data Requirement Been Addressed? Source Number or Other 

Number How? Reference 

Yes. Data waiver Previously submitted 

885.4280 Estuarine and marine rationale in MRID U.S. EPA (2008b) data are adequate to 

invertebrate testing fulfill data requirement 471572021 

by bridging. 

B. sphaericus is not 

885.4300 Nontarget plant Yes. Known insect U.S. EPA (2008b) taxonomically related to 

Testing pathogen a known plant 

pathogen. 

885.4340 Nontarget insect testing Yes. Data waiver Previously submitted 

rationale in MRID U.S. EPA (2008b) data are adequate to 

885.4380 Honey bee testing 471572021 fulfill data requirements 

by bridging .. 1U.S.EPA. 2008c, 2011 e, 2011f- s1m1lanty shown to ABTS 1743. Request 1n MRID 47157202 to wa1ve test1ng relied on demonstration of 

similarity to ABTS 1743 (see product chemistry Table 4) and citation to previously submitted data. 2U.S. EPA 2008b: Memorandum from 

S. Borges to M. Mendelsohn. dated 09/02/08. 

E. Risk to Threatened and Endangered Species 

EPA has prepared risk assessments that support a complete endangered species 

determination for the registered Bacillus sphaericus strains ABTS 1743 and AM614 (U.S. EPA, 

Z. Vaituzis to P. Hutton, 08/04/1988; S. Borges toM. Mendelsohn U.S. EPAIBPPD DER dated 

09/02/2008). During Registration Review, EPA plans to conduct an ecological risk assessment 

to reassess whether use of the registered Bacillus sphaericus strains will have "no effect" or 

"may affect" federally listed threatened or endangered species (listed species) and/or their 

designated critical habitat. When an assessment concludes that a pesticide's use "may affect" 

a listed species and/or its designated critical habitat, EPA will consult with the U.S. Fish and 

Wildlife Service and/or National Marine Fisheries Service (the Services), as appropriate. 

F. Product Performance 

Product Performance data submitted for Bacillus sphaericus ABTS 1743 and AM614 

demonstrate that the range of efficacy for EPs tested in acceptable studies range from 85 to 

1 00%. (U.S. EPA 1999, 2006, 2008a, 2008e, 2011 g, 2012). 

G. Endocrine Effects 

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 in order 

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 Estrogenic Substances Screening Program a biologic substance or other 

19 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www.regulations.gov 

substance if a determination is made that the substance is not anticipated 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 contains 58 pesticide active ingredients and 9 inert ingredients. 

The Bacillus sphaericus strains ABTS 1743 and AM614 are not among the group of 58 

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

EPA, as part of this Preliminary Work Plan, believes that the Bacillus sphaericus strains ABTS 

1743 and AM614, the active ingredients involved in this Registration Review case, likely are 

substances that would not produce any effect in humans similar to an effect produced by a 

naturally occurring estrogenic substance. As such, pursuant to Section 408(p)(4), EPA will 

determine in the future whether it can exempt the Bacillus sphaericus strains ABTS 1743 and 

AM614 from the requirements of the Section 408(p) EDSP. In the event EPA does determine 

to exempt these substances 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 at http://www.epa.gov/endo/. 

H. Incidents 

There were no incidents for Bacillus sphaericus pesticides in the Environmental Incident 

Information System (EllS) database up to the last update in August 2011. In accordance with 

section 6(a)(2), VBS reported two incidents over the two decades of use of ABTS 1743. One 

was a human minor (HD) in 1991 and the other was a moderate domestic animal report in 

2003. As of March 2013, these were reported as line items in a quarterly aggregate report of 

incidents in OPP's Incident Data System (IDS); no detailed information was included in the 

main individual study reports that are part of OPP's IDS. 

I. Timeline 

The projected timeline for case 6052 - Bacillus sphaericus - Registration Review follows. 

Activities Estimated MonthNear 

Opening the Docket 

Open Docket and 60-Day Public Comment Period for Bacillus March 27, 2013 

sphaericus 

Close Public Comment Period May 27, 2013 

Case Development 

Issue Final Work Plan September 2013 

Issue Data Call-in January 2015 

Data Submission January 2016 

Open 30-Day Public Comment Period for Draft Risk Assessments October 2016 

Close Public Comment Period November 2016 

Registration Review Decision 

20 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations. gov 

Activities Estimated MonthNear 

Open 60-Day Public Comment Period for Proposed Registration March 2018 

Review Decision 

Close Public Comment Period May 2018 

Final Decision September 2018 

*Estimated Total (years) 4.5 .. * Th1s schedule IS subject to rev1s1on should unforeseen 1ssues anse dunng the reg1strat1on rev1ew process. In the 

event an issue arises, such as the failure to acquire an EDSP exemption, an amended Final Work Plan will be 

issued at that time. That Final Work Plan will identify any newly anticipated data needs and set forth a new 

timeline. 

J. Guidance for Commenters 

The public is invited to comment on EPA's Preliminary Work Plan and rationale. The areas 

below highlight topics of special interest to EPA where comments, information, submissions, or 

reference to sources of additional information could be of particular use. EPA will consider all 

comments, as well as any additional information or data provided in a timely manner, prior to 

issuing a Final Work Plan for this case. 

i. 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, in the development, 

implementation, and enforcement of environmental laws, regulations, and policies. To help 

address potential environmental justice issues, EPA seeks information on any groups or 

segments of the population who, as a result of their location, cultural practices, or other factors, 

may have atypical, unusually high exposure to these registered pesticides containing Bacillus 

sphaericus strains, 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. 

ii. Water Quality 

The Bacillus sphaericus strains in registered pesticide products are not identified as a cause of 

impairment for any water bodies listed as impaired under section 303(d) of the Clean Water 

Act. 2 In addition, no Total Maximum Daily Loads (TMDLs) have been developed for these 

Bacillus sphaericus strains: ABTS 17 43 and AM614.3 More information on impaired water 

bodies and TMDLs can be found at EPA's website.5 EPA invites submission of water quality 

data for these pesticides. To the extent possible, data should conform to the quality 

standards in Appendix A of the Office of Pesticide Program's (OPP) Standard Operating 

Procedure: Inclusion of Impaired Water Body and Other Water Quality Data in OPP's 

Registration Review Risk Assessment and Management Process4 to ensure they can be used 

quantitatively or qualitatively in pesticide risk assessments. 

2 Based on information provided at http:/lwww.epa.gov/owowltmdl/. 

3 Based on information provided at http://iaspub.epa.gov/tmdl waterslO/text search.tmdl search form. 5 See http:/lwww.epa.gov/owowltmdll. 4 This document can be found at http://www.epa.gov/oppsrrdllregistration review/water quality sop.htm. 

21 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations.gov 

iii. Trade Irritants 

Through the Registration Review process, EPA intends to solicit information on trade irritants 

and, to the extent feasible, take steps toward facilitating irritant resolution. EPA will work to 

harmonize tolerance and international Maximum Residue Limits (MRLs) and may modify 

tolerance levels to do so, when possible. Growers and other stakeholders are asked to 

comment on any trade irritant issues resulting from lack of MRLs or disparities between U.S. 

tolerances and MRLs in key export markets, providing as much specificity as possible 

regarding the nature of the concern. 

An exemption from the requirement of a tolerance has been established in the U.S. for Bacillus 

sphaericus in 40 CFR 180.1202. Moreover, there are no known MRLs for Bacillus sphaericus 

strains: ABTS 17 43 and AM614. EPA does not anticipate that current uses of Bacillus 

sphaericus strains: ABTS 17 43 and AM614 will pose concerns as trade irritants. 

iv. Additional Information 

Stakeholders are also specifically asked to provide information and data that will assist EPA in 

refining the risk assessments. EPA is interested in obtaining the following information regarding 

the Bacillus sphaericus strains, ABTS 17 43 and AM614: 

~ Confirmation on the following label information: 

- Sites of application 

- Formulations 

-Application methods and equipment 

- Maximum application rates 

- Frequency of application, application intervals, and maximum number of 

applications 

-Geographic limitations on use 

~ Use or potential use distribution (e.g., acreage and geographical distribution of 

relevant use sites); 

~ Median and goth percentile reported use rates (lbs active ingredient/1 ,000 square 

feet) from usage data- national, state, and county; 

~Application timing (date of first application and application intervals)- national, state, 

and county; 

~ Usage/use information for nonagricultural uses; 

~Typical application interval (days); 

~ State or local use restrictions; 

~ Monitoring data. 

K. Next Steps 

After the 60-day comment period closes, EPA will review and respond to any comments 

received in a timely manner and then issue a Final Work Plan for case 6052: Bacillus 

sphaericus. 

22 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations. gov 

II. FACT SUMMARY FOR CASE 6052 

TABLE 11. Background Information for Bacillus sphaericus Registration Review 

Publicly Available Additional Risk Assessmenf. 3 

Active Ingredient First Currently Registered EPA Science and Needed? (Yes or No) 

(PC Code) Registered Pesticide Products' Regulatory Product Human Ecological Documents Chemistry HeaHh 

Bacillus 1991 1 MP( 73049-57) registered Registration Notices 

sphaericus strain 2001 and Labels on 

2362 serotype 3 end-use products www. epa.gov/pesticides/ 

H5a5b ABTS1743 (73049-20, 73049-57, 73049- ppls 

(PC Codes 429) Citations in No No Yes 

119801, 119803) Bibliography and 

references to EPA 

reviews in this 

document. 

Bacillus 1991 1 MP (84268-1) and 2 EPs 3 Registration Notices 

sphaericus strain (84268-2, 84268-3). and Labels on 

2362 serotype www.epa.gov/pesticides/ 

H5a5b AM614 Three additional EPs3 ppls 

(PC Codes (83362-3, 85685-2, 85685-3) Citations in No No Yes 

119801, 119804) contain AM614. Bibliography and 

references to EPA 

reviews in this 

document. 0 0 1 See Table 1 for add1t1onal1nformat1on (e.g., use s1tes) on the currently registered products. Labels for these products can be found 1n the 

Pesticide Product Label System (www.epa.gov/pesticides/ppls). 2See Anticipated Data Needs and Risk Assessments. 

3Product registrations is an ongoing process and subject to reviews as data/information are submitted. 

23 
Bacillus splwericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations.gov 

Ill. BIBLIOGRAPHY 

A. Studies Supporting Registration Review of Bacillus sphaericus strain ABTS 17 43 (PC 

Codes119801, 119803) 

MRID 

32514 

148466 

148467 

148468 

148469 

148470 

148471 

156631 

156632 

156633 

156634 

40028000 

40028001 

40028002 

40028003 

40193700 

40193701 

40329800 

Bibliography: Bacillus sphaer/cus 2362 serotype H5a5b ABTS 1743 

(PC code119801, 119804) 

Citation Reference 

Goldberg, L.J.; Margalit, J. (1977) A bacterial spore demonstrating rapid larvicidal activity against 

Anopheles sergentii, Uranotaenia unguiculata, Culex univitattus, Aedes aegypti, and Culex pipiens. 

Mosquito News 37(3):355-358. (Also In unpublished submission received Apr 15, 1980 under 

43382-3; submitted by Biochem Products, S.A., Brussels, Belgium; CDL: 242360-J). 

Biochem Products S.C.S. (1985) BSP 1: Chemical and Physical Properties. Unpublished study.1 p. 

Biochem Products S.C.S. (1985) Toxicity Data or Reference to Available Data on the Toxicity of the 

Pesticide (Bacillus sphaericus) Including Data on the Toxicity to Fish and Wildlife. Unpublished 

study. 5 p. 

De Barjac, H.; Larget, 1.; Cosmao, V.; et al. (19??) lnnocuite de Bacillus sphaericus, Souche 1593, 

pour les Mammiferes: WHONBC/79.731. Unpublished study prepared by World Health 

Organization. 20 p. 

World Health Organization, Div. of Vector Biology and Control (1980) Data Sheet on the Biological 

Control Agent: Bacillus sphaericus, Strain 1593: HONBC/80. 777: VBC/BCDS/80.1 0. Unpublished 

study. 15 p. 

Shadduck, J. (1983) Some considerations on the safety evaluation of nonviral microbial pesticides. 

Bulletin of the World Health Organization 61 (1):117-128. 

Biochem Products S.C.S. (1985) Effectiveness Data Which Indicate That the Product (B.S.P. 1) Is 

Useful for the Purposes Proposed. Unpublished compilation. 28 p. 

Abbott Laboratories (1986) Chemical and Physical Properties: [Bacillus sphaericus-Biological 

Mosquito Larvicide]. Unpublished study. 4 p. 

Shadduck, J.; Singer, S.; Lause, S. (1980) Lack of mammalian pathogenicity of entomocidal 

isolates of Bacillus sphaericus. Entomological Society of America 9(4):403-407. 

Shadduck, J. (1985) Mammalian Safety of Bacillus sphaericus: BCV/ CSP/WP.25. Unpublished 

review of studies prepared by World Health Organization. 6 p. 

Mulla, M. (1985) Safety of Bacillus sphaericus to Beneficial Macro- invertebrates: BCV/CSP/WP.24. 

Unpublished study prepared by World Health Organization. 6 p. 

Abbott Laboratories (1986) Product Chemistry of Bacillus sphaericus H-5a5b Granules. 

Compilation of 3 studies. 

Devissetty, B. (1986) Product Identity and Composition of[ ... ] Bacillus sphaericus H-5a5b 

Granules. Unpublished compilation prepared by Abbott Laboratories. 39 p. 

Devisetty, B. (1986) Analysis and Certifications of Product Ingredients of Bacillus sphaericus H5a5b Granules. Unpublished study prepared by Abbott Laboratories. 15 p. 

Devisetty, B. (1986) Physical/Chemical Properties of[ ... ] Bacillus sphaericus H-5a5b Granules. 

Unpublished study prepared by Abbott Laboratories. 11 p. 

Abbott Laboratories (1987) Submission of Toxicity Data in Support of Application for Registration of 

Bacillus sphaericus H-5a5b. Transmittal of 1 study. 

Foster, R. (1987) Supplementary Information on the Toxicity of Bacillus sphaericus H-5a5b to Nontarget Aquatic Organisms. Unpublished study prepared by Abbott Laboratories. 8 p. 

Abbott Laboratories (1987) Submission of Toxicity Data To Support the Registration of Bacillus 

24 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations. gov 

MRID 

40329801 

40329802 

40329803 

40329804 

40329805 

40329806 

40329807 

40329808 

40329809 

40329810 

40329811 

40329812 

40329813 

40329814 

40329815 

40329816 

40391400 

Bibliography: Bacillus sphaericus 2362 serotype H5a5b ABTS 1743 

(PC code119801, 119804) 

Citation Reference 

sphaericus H-5a5b Granules. Transmittal of 16 studies. 

Cozzi, E. (1986) An Overview of the Toxicity and Pathogenicity of Bacillus sphaericus H-5a5b to 

Mammals: Document No.4. Unpublished compilation prepared by Abbott Laboratories. 53 p. 

Henck, J. (1986) Bacillus sphaericus: Acute Oral LD50 in Rats: Document No.5: TRL Study No. 

024-025. Unpublished study pre- pared by Toxicity Research Laboratories, Ltd. 25 p. 

Henck, J. (1986) Bacillus sphaericus: Acute Dermal Toxicity Study: Document No. 7: TRL Study 

No. 024-026. Unpublished study pre- pared by Toxicity Research Laboratories, Ltd. 27 p. 

Newton, P. (1986) Four Hour Acute Dust Aerosol Inhalation Toxicity Study in Rats of Bacillus 

sphaericus: Document No.9: ABC Study No. 420-2507. Unpublished study prepared by American 

Biogenics Corp. 24 p. 

Henck, J. (1986) Intravenous Toxicity/Infectivity Study with Bacillus sphaericus in Newly Weaned 

Mice: Document No. 11: TRL Study No. 024-029. Unpublished study prepared by Toxicity 

Research Laboratories, Ltd. 32 p. 

Henck, J. (1986) Intravenous Toxicity/Infectivity Study with Bacillus sphaericus in Newly Weaned 

Hamsters: Document No. 12: TRL Study No. 024-028. Unpublished study prepared by Toxicity 

Research Laboratories, Ltd. 24 p. 

Henck, J. (1986) Bacillus sphaericus: Primary Dermal Irritation Study: Document No. 13: TRL 

Study No. 024-023. Unpublished study prepared by Toxicity Research Laboratories, Ltd. 28 p. 

Henck, J. (1986) Bacillus sphaericus: Primary Eye Irritation Study: Document No. 14: TRL Study 

No. 024-024. Unpublished study pre- pared by Toxicity Research Laboratories, Ltd. 33 p. 

Henck, J. (1986) Hypersensitivity Study of Bacillus sphaericus in Albino Guinea Pigs: Document 

No. 15: TRL Study No. 024-027. Unpublished study prepared by Toxicity Research Laboratories, 

Ltd. 48 p. 

Cozzi, E. (1986) An Overview of the Toxicity and Pathogenicity of Bacillus sphaericus to Non-target 

Organisms: Document No. 16. Unpublished study prepared by Abbott Laboratories. 8 p. 

Henck, J. (1986) Bacillus sphaericus: Avian Single Dose Oral Toxicity Study in Mallard Ducks: 

Document No. 17: TRL Study No. 024-030. Unpublished study prepared by Toxicity Research 

Laboratories, Ltd. 17 p. 

Henck, J. (1986) Bacillus sphaericus: Avian Injection Pathogenicity Test in Mallard Ducks: 

Document No. 18: TRL Study No. 024-031. Unpublished study prepared by Toxicity Research 

Laboratories, Ltd. 25 p. 

Surprenant, D. (1986) Bacillus sphaericus: Acute Toxicity of ABG - 6184 to Bluegill (Lepomis 

machrochirus): Document No. 19: Study No. 2439.0786.6100.100. Unpublished study prepared by 

Spring- born Bionomics, Inc. 22 p. 

Surprenant, D. (1987) Bacillus sphaericus: Acute Toxicity of ABG- 6184 to Rainbow Trout (Salmo 

gardnen) (sic): Document No. 20: Study No. 2439.0786.6100.103. Unpublished study prepared by 

Springborn Bionomics, Inc. 21 p. 

Surprenant, D. (1986) Bacillus sphaericus: Acute Toxicity of ABG- 6184 to Daphnids (Daphnia 

magna): Document No. 21: Study No. 2439.0786.61 00.11 0. Unpublished study prepared by 

Springborn Bionomics, Inc. 22 p. 

Vandenberg, J. (1986) Safety of Bacillus sphaericus for Caged Adult Honey Bees: Document No. 

24. Unpublished study prepared by USDA-ARS Beneficial Insects Laboratory. 12 p. 

Abbott Laboratories (1987) Submission of Toxicity Data To Support the Application for the 

Registration of Bacillus sphaericus Serotype H-5a5b Biological Larvicide. Transmittal of 1 study. 

25 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations. gov 

MRID 

40391401 

40429500 

40429501 

40885300 

40885301 

40885302 

40886600 

40886601 

40886602 

41103400 

41103401 

41103402 

41103403 

41586701 

41586702 

41586703 

41604000 

41604001 

41931300 

Bibliography: Bacillus sphaericus 2362 serotype H5a5b ABTS 1743 

(PC code119801, 119804) 

Citation Reference 

Surprenant, D. (1986) Bacillus sphaericus: Acute Toxicity of ABG- 6184 to Mayfly Larva 

(Hexagenia sp.): Study No. 2439.0786.6100. 711. Unpublished study prepared by Springborn 

Bionomics, Inc. 22 p. 

Abbott Laboratories (1987) Submission of Supplementary Information in Response to Preliminary 

Review Comments on B. sphaericus H-5a 5b Granules. Transmittal of 1 study. 

Jovanovich, A.; Cozzi, E. (1987) Application to Register B. sphaericus H-5a5b Granules for Control 

of Mosquito Larvae. Unpublished compilation prepared by Technology Services Group, Inc. 74 p. 

Abbott Laboratories (1988) Submission of Chemistry Data in Support of Vectolex G Powder; 

Granular Biological Mosquito Larvicide Containing Bacillus sphaericus. Transmittal of 2 studies. 

Rowell, R. (1988) Vectolex G: Product Chemistry. Unpublished compilation prepared by Abbott 

Laboratories. 25 p. 

Melin, B.; Coddens, M. (1988) Vectolex G: Product Chemistry. Unpublished compilation prepared 

by Abbott Laboratories. 23 p. 

Abbott Laboratories (1988) Submission of Data To Support Registration of Vectolex Tech Powder: 

Product Chemistry Data. Transmittal of 2 studies. 

Smith, R.; Cooper, R. (1988) Vectolex Technical: Product Chemistry: 10/88-01. Unpublished study 

prepared by Abbott Laboratories. 240 p. 

Melin, B.; Coddens, M. (1988) Vectolex Technical: Product Chemistry: 10/88-02. Unpublished 

study prepared by Abbott Laboratories. 23 p. 

Abbott Laboratories (1989) Submission of Toxicity Data in Support of Bacillus sphaericus 

Registration Standard. Transmittal of 3 studies. 

David, R. (1989) Acute Oral Toxicity/Pathogenicity Study of Bacillus sphaericus Technical Powder 

in Rats: Final Report: Project No. G-7161.222. Unpublished study prepared by Microbiological 

Associates Inc. 21 p. 

David, R. (1989) Acute Pulmonary Toxicity/Pathogenicity Study of Bacillus sphaericus Technical 

Powder in Rats: Project No.G-7161. 225. Unpublished study prepared by Microbiological 

Associates Inc. 21 p. 

David, R. (1989) Acute Intravenous Toxicity/Pathogenicity Study of Bacillus sphaericus Technical 

Powder in Rats: Project No. G-7161 224. Unpublished study prepared by Microbiological 

Associates Inc. 21 p. 

Eliezer, E. (1989) Product Chemistry Based on Bacillus sphaericus, (ABTS-1743) as the Active 

Ingredient: Lab Project Number: BSP 7/90-02. Unpublished study prepared by Abbott Laboratories. 

42p. 

Jaronski, S. (1990) Physical Stability Study of Technical Grade Bacillus sphaericus: Lab Project 

Number: 910-8903. Unpublished study prepared by Abbott Laboratories. 26 p. 

Smith, R. (1990) Product Identity and Disclosure of Ingredients: Bacillus sphaericus: Lab Project 

Number: BSP 7/90-04. Unpublished study prepared by Abbott Laboratories. 10 p. 

Abbott Laboratories (1990) Submission of Toxicity/Pathogenicity Data in Support of Bacillus 

sphaericus Registration. Transmittal of 1 study. 

Jaronski, S. (1990) Host Spectrum Study of Technical Grade Bacillus sphaericus: Lepidoptera, 

Orthoptera, and Coleoptera: Lab Project Number: 91 0-8908. Unpublished study prepared by Abbott 

Laboratories. 20 p. 

Abbott Laboratories (1991) Submission of Data To Support Registration of Vectolex, Biological 

26 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations. gov 

MRID 

41931301 

41931302 

41931303 

41931304 

41931305 

43016200 

43016201 

43016202 

43991900 

43991901 

44216500 

44216501 

44572310 

44631700 

44631701 

45233000 

Bibliography: Bacillus sphaericus 2362 serotype H5a5b ABTS 1743 

(PC code119801, 119804) 

Citation Reference 

Mosquito Larvicide: Toxicology Studies. Transmittal of 5 studies. 

Bowman, J. (1989) Acute Toxicity of Bacillus sphaericus Technical Material to Sheepshead Minnow 

(Cyprinodon variegatus) in a Static Renewal System: Lab Project Number: 37730. Unpublished 

study prepared by Analytical Biochemistry Labs., Inc. 93 p. 

Forbis, A. (1990) Acute Toxicity of Bacillus sphaericus to Mysidopsis bahia: Final Report: Lab 

Project Number: 38126. Unpublished study prepared by Analytical Bio-chemistry Laboratories, Inc. 

98 p. 

Forbis, A. (1989) Acute Toxicity of Bacillus sphaericus to Se/enastrum capricornutum Printz: Final 

Report: Lab Project No: 37657. Unpublished study prepared by Analytical Bio-chemistry 

Laboratories, Inc. 80 p. 

Grimes, J. (1989) Bacillus sphaericus Technical Powder: An Avian Dietary Pathogenicity and 

Toxicity Study in the Mallard: Lab Project Number: 161-111. Unpublished study prepared by 

Wildlife International Ltd. 41 p. 

Dionne, E. (1990) Bacillus sphaericus Technical Material--Acute Toxicity to Eastern Oysters 

( Crassostrea virginica) Under Flow- through Conditions: Final Report: Lab Project No: 89-06-3017; 

2439.0289.6102.504. Unpublished study prepared by Springborn Laboratories, Inc. 45 p. 

Abbott Labs. (1993) Submission of Product Chemistry Data for ABG-3097 in Support of 

Registration. Transmittal of 2 Studies. 

Rojas, F. (1993) Addendum to Report Titled Physical and Chemical Properties Characterization of 

ABG-3097 Technical Grade Active Ingredient and End-use Material--MAID No. 42704701: Lab 

Project Number: 81-2389-62. Unpublished study prepared by Abbott Labs., Chemical & Agricultural 

Products Div. 59 p. 

Mathieu, G. (1993) Storage Stability and Corrosion Characteristics of ABG-3097 Technical Grade 

Active Ingredient and End-use Material: Final Report: Project No. 69-2002-62. Prepared by Abbott 

Labs. 23 p. 

Abbott Laboratories (1996) Submission of Toxicology Data in Support of the Amended Registration 

of Vectolex CG Biological Larvicide. Transmittal of 1 Study. 

Kuhn, J. (1996) Primary Dermal Irritation Study in Rabbits: VectoLex Technical Powder: Final 

Report: Lab Project Number: 2709-96: S9-FF81-5.A 12. Unpublished study prepared by 

Stillmeadow, Inc. 16 p. 

Abbott Laboratories (1997) Submission of Efficacy Data in Support of the Registration for Vectolex 

CG Biological Larvicide. Transmittal of 1 Study. 

Melin, B. (1997) Field/Laboratory Performance Evaluations of Bacillus sphaericus: Lab Project 

Number: BS EF-1. Unpublished study prepared by Abbott Laboratories. 120 p. 

Melin, B. (1998) Field Evaluations of Vectolex WDG, ABG-6491: Lab Project Number: AB019: BS 

WDG EF-1. Unpublished study prepared by Benzon Research. 58 p. 

Abbott Laboratories (1998) Submission of Efficacy Data in Support of the Registration of Vectolex 

CG Biological Larvicide. Transmittal of 1 Study. 

Melin, B. (1997) Efficacy of Vectolex CG and Vectolex WDG against Larvae of Coquilletidia 

perturbans in a Cattail Marsh in Capemay, NJ: Lab Project Number: AB012. Unpublished study 

prepared by Benzon Research. 8 p. 

Valent Biosciences (2000) Submission of Product Chemistry Data in Support of the Application for 

the Registration of Bacillus thuringiensis subsp. aizawai Slurry, Bacillus subsp. israe/ensis Slurry, 

Bacillus subsp. kurstaki Slurry, Bacillus subsp. tenebrionis Slurry and Bacillus subsp. sphaericus 

27 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations. gov 

MRID 

45233004 

45681800 

45681801 

46511000 

46511001 

46724405 

46724406 

46724407 

Bibliography: Bacillus sphaericus 2362 serotype H5a5b ABTS 1743 

(PC code119801, 119804) 

Citation Reference 

Slurry and Myrothecium verrucaria Slurry. Transmittal of 2 Studies. 

Rowell, R. (2000) Slurry Manufacturing Process for Vectolex Products (Bacillus sphaericus) at 

Abbott Laboratories: Lab Project Number: BSSLMAN-V28R. Unpublished study prepared by Abbott 

Laboratories. 27 p. 

Valent BioSciences Corp. (2002) Submission of Product Chemistry Data in Support of the 

Application for Registration of Bacillus sphaericus Slurry. Transmittal of 1 Study. 

Rowell, R. (2002) Slurry Manufacturing Process for Vectolex Products (Bacillus sphaericus) at 

Abbott Laboratories: Lab Project Number: 60012 MAN 5/21/02: SCP.V.78339. Unpublished study 

prepared by Abbott Laboratories. 27 4 p. {OPPTS 885.1200} 

Valent Biosciences Corporation (2005) Submission of Product Chemistry Data in Support of the 

Amended Registration of VectoLex CG Biological Larvicide Granules. Transmittal of 1 Study. 

Woolley, A.; Mullee, D. (2005) Determination of Dust Content and Friability and Attrition: ABG6185. Project Number: 1438/002. Unpublished study prepared by Safepharm Laboratories Ltd. 9 p. 

Durando, J. (2005) Dermal Sensitization Study in Guinea Pigs (Buehler Method): VBC-60034. 

Project Number: 17961, P328. Unpublished study prepared by Product Safety Laboratories. 28 p. 

DeChant, P. (2005) Field Efficacy Data for VBC-60035. Project Number: VBC/12/09/05/1, 

2002PDECH026, 2004PPDECH024. Unpublished study prepared by Valent Biosciences 

Corporation. 1 0 p. 

Smith, W. (2005) Discussion of Requirements for Biochemical Pesticide Toxicity Data: V60035 CG. 

Project Number: D/928NBC/60035. Unpublished study prepared by Valent Biosciences 

Corporation. 36 p. 

B. Studies Supporting Registration Review of Bacillus sphaericus serotype H5a5b strain AM614 (PC 

code 119801, 119804) 

MRID 

Bibliography: Bacillus sphaericus 2362 serotype H5a5b AM614 

(PC code119801, 119804) 

Citation Reference 

84268-1- Advanced Microbiologies, LLC 

47157200 Advanced Microbiologies, LLC (2007) Submission of Product Chemistry, Efficacy, and Toxicity 

Data in Support of the Application for Registration of AM61432 Primary Powder. Transmittal of 10 

Studies. 

47157201 

47157202 

47157203 

47157204 

Roberts, A. (2007) Product Chemistry for AM61432 Primary Powder. Unpublished study prepared 

by Technology Sciences Group, Inc. 119 p. 

Mileson, B. (2007) Response to Tier 1 Microbial Pesticide Data Requirements for AM61432 

Primary Powder. Unpublished study prepared by Technology Sciences Group, Inc. 66 p. 

Moore, G. (2005) Acute Oral Toxicity Up and Down Procedure in Rats: Bacillus sphaericus Strain 

2362. Project Number: 17860, P320/UDP. Unpublished study prepared by Product Safety 

Laboratories. 15 p. 

Moore, G. (2005) Acute Dermal Toxicity Study in Rats- Limit Test: Bacillus sphaericus Strain 2362. 

Project Number: 17861, P322. Unpublished study prepared by Product Safety Laboratories. 15 p. 

28 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations. gov 

MRID 

47157205 

47157206 

47157207 

47157208 

47157209 

47157210 

47356800 

47356801 

47898200 

47898201 

47898202 

48391600 

48391601 

48405900 

48405901 

Bibliography: Bacillus sphaericus 2362 serotype H5a5b AM614 

(PC code119801, 119804) 

Citation Reference 

Moore, G. (2005) Acute Inhalation Toxicity Study in Rats- Limit Test: Bacillus sphaericus Strain 

2362. Project Number: 17862, P330. Unpublished study prepared by Product Safety Laboratories. 

23 p. 

Moore, G. (2005) Primary Eye Irritation Study in Rabbits: Bacillus sphaericus Strain 2362. Project 

Number: 17863, P324. Unpublished study prepared by Product Safety Laboratories. 15 p. 

Moore, G. (2005) Primary Skin Irritation Study in Rabbits: Bacillus sphaericus Strain 2362. Project 

Number: 17864, P326. Unpublished study prepared by Product Safety Laboratories. 15 p. 

Moore, G. (2005) Dermal Sensitization Study in Guinea Pigs (Buehler Method): Bacillus sphaericus 

Strain 2362. Project Number: 17865, P328. Unpublished study prepared by Product Safety 

Laboratories. 24 p. 

Hicks, S. (2007) Bacillus sphaericus Strain 2362: Tier I Static Renewal 21-Day Toxicity Test with 

Daphnia magna. Project Number: 50395. Unpublished study prepared by ABC Laboratories, Inc. 

55 p. 

Becnel, J. (2007} Bacillus sphaericus Strain 2362: Field and Lab Mortality Tests. Unpublished study 

prepared by USDA, ARS, Center for Medical Agricultural and Veterinary Entomology. 20 p. 

Advanced Microbiologies, LLC (2008) Submission of Product Chemistry and Efficacy Data in 

Support of the Applications for Registration of AM61432 Primary Powder, AM61432 (50 G) and 

AM61432 WP. Transmittal of 3 Studies. 

Roberts, A. (2008} Supplemental Product Chemistry for AM61432 Primary Powder. Project 

Number: 07584D/GB/2002/10. Unpublished study prepared by Technology Science Group, Inc. 36 

p. 

Advanced Microbiologies, LLC. (2009} Submission of Product Chemistry Data in Support of the 

Registration of AM61432 Primary Powder. Transmittal of 2 Studies. 

Federici, B. (2009) Supplemental Product Identity for AM61432 Primary Powder: Final Report. 

Unpublished study prepared by University of California, Riverside. 71 p. 

Roberts, A. (2009} Supplemental Product Chemistry for AM61432 Primary Powder: Final Report. 

Unpublished study prepared by Technology Sciences Group, Inc. 20 p. 

Advanced Microbiologies, LLC (2011) Submission of Product Chemistry Data in Support of the 

Registration of AM61432 Primary Powder. Transmittal of 1 Study. 

Roberts, A. (2011) Supplemental Product Chemistry for AM61432 Primary Powder. Unpublished 

study prepared by Advanced Microbiologies, LLC. 28 p. 

Advanced Microbiologies, LLC (2011) Submission of Product Chemistry Data in Support of the 

Registration of AM61432 Primary Powder. Transmittal of 1 Study. 

Roberts, A. (2011) Supplemental Product Chemistry for AM61432 Primary Powder. Unpublished 

study prepared by Technology Science Group, Inc. 28 p. 

C. Environmental Protection Agency Risk Assessment Memoranda 

U.S. EPA. 1988. EEB Review: Ecological effects Bacillus sphaericus H5a5b granules mosquito larvicides. 

Memorandum from Z. Vaituzis to P. Hutton dated August 4, 1988. 

U.S. EPA. 1989a. SACB Review: Bacillus sphaericus mosquito larvicide. Abbott Laboratories' registration 

application for Vectolex technical and granular products. Memorandum from W. J. Hazel toP. Hutton 

dated March 22, 1989. 

29 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations. gov 

U.S. EPA. 1989b. SACB review of Toxicity Studies submitted by Abbott Laboratories to support registration of 

Bacillus sphaericus H5a5b granules. Memorandum from J. T. McClintock toP. Hutton dated September 

14, 1989. 

U.S. EPA. 1991. SACB Review of Additional Product Identity Information Submitted by Abbott for Bacillus 

sphaericus (Vectolex TGAI and Vectolex G). [EPA ID No. 275-TT; Record No. 381936; MRID Nos. 

41586701, 02, and 03; Caswell No. 066A; H ED Project No. 0-2053) 

U.S. EPA. 1996a. Horne, D. Acute Toxicity of Bacillus sphaericusTechnical Material to Sheepshead Minnow 

(Cyprinodon variegatus) in a Static Renewal System (Guideline 72-1/885.4280) Data Evaluation Record 

dated August 19, 1996. MAID 41931301. 

U.S. EPA. 1996b. Horne, D. Acute Toxicity of Technical Material Bacillus sphaericus to Mysidopsis bahia 

(Guideline 72-3/885 .4280). Data Evaluation Record dated August 19, 1996. MRID 41931302. 

U.S. EPA. 1996c. Horne, D. Acute Toxicity of Bacillus sphaericus to Selanastrum capricornutum Printz- static 

Algal Bioassay. (Guideline 122-2/850.5400). Data Evaluation Record dated August 19, 1996. MRID 

41931303. 

U.S. EPA. 1996d. Horne, D. Acute Toxicity of Bacillus sphaericus Technical Material to crassostrea virginica 

(Eastern Oyster)- Shell Deposition Bioassay (Guideline 154A-21/ 885.4280)Data Evaluation Record 

dated August 19, 1996. MRID 41931305. 

U.S. EPA. 1996e. BPPD Review of Primary Dermal Irritation Study for Technical B.sphaericus. J. F. Heminway 

dated November 24, 1996. 

U.S. EPA. 1999b. BPPD Review of Field and laboratory efficacy studies to support product labeling claims to 

control mosquitoes- Vectolex WDG. Memorandum from Robyn Rose toW. Nelson dated January 7, 

1999. 

U.S. EPA. 2001: BPPD Review of Data Submitted by Valent BioSciences Corporation, for Registration and 

Review of Manufacturing (Use Product) Production Process to Vectolex Products (Bacillus sphaericus) at 

Abbott Laboratories. Memorandum from C. Etsitty to A. Reynolds dated October 15, 2001. 

U.S. EPA. 2004. BP1 D Review of Data Submitted by Valent BioSciences Corporation, for Registration and 

Review of Manufacturing (Use Product)Production Process to Vectolex Products (Bacillus sphaericus) at 

Abbott Laboratories. Memorandum from C. Etsitty to A. Reynolds dated September 22, 2004. 

U.S. EPA. 2006. BPPD Review of Product Identity, Manufacturing Process, Formation of Unintentional 

Ingredients, Analysis of Samples, Certification of Limits, Physical & Chemical Characteristics, and Five 

batch analyses, Efficacy, Storage Stability, Acute Oral Toxicity, Dermal Sensitization and Primary Eye 

Irritation Studies of the End Use Product: V60035 CG Biological Larvicide to determine its eligibility for 

registration. Memorandum from I. Barsoum to A. Reynolds dated April 10, 2006. 

U.S. EPA. 2008a. BPPD Review of Product Performance Data to Support Registration of the New Products 

AM61432 (50G) (EPA File Symboi84268-E) and AM61432 WP (EPA File Symbol 84268-G) Containing 

Bacillus sphaericus, Serotype H5a5b, strain 2362 (PC Code 119801 ); Submitted by Advanced 

Microbiologies, LLC for AM61432 WP. Memorandum from S. Borges toM. Mendelsohn dated August 29, 

2008. 

U.S. EPA. 2008b. Ecological Risk Assessment for the Active Ingredient Bacillus sphaericus H5a5b strain 2362 to 

Support Registration of the New Product AM6 1432 Primary Powder (EPA File Symbol 84268-R); 

Submitted by Advanced Microbiologies, LLC. Memorandum from S. Borges toM. Mendelsohn dated 

September 02, 2008. 

U.S. EPA. 2008c. Product Chemistry, Manufacturing Process and Acute toxicities studies for AM61432 Primary 

Powder (EPA Reg. No. 84268-R). Memorandum from I. Barsoum toM. Mendelsohn dated September 09, 

2008. 47157201, 47356801 (Pchem) 47157203 to 08- 03 oral, 04 dermal, 05 inhalation, 06 eye, 07 skin 

irritation, 08 dermal sensitization- Pasteur deposit. 

U.S. EPA. 2008d. BPPD Ecological Risk Assessment for the Active Ingredient Bacillus sphaericus HSaSb strain 

2362 to Support Registration of the New Product AM61432 Primary Powder. Memorandum from S. 

Borges toM. Mendelsohn dated October 29, 2008. 

30 
Bacillus sphaericus Case 6052- Preliminary Work Plan EPA-HQ-OPP-2013-0116 www. regulations.gov 

U.S. EPA. 2008e. BPPD Review of Product Performance Data to Support Registration of the New Product 

FourStar Briquets (EPA File Symbol 83362-G) Containing Bacillus sphaericus, Serotype H5a5b, strain 

2362 (PC Code 119801) and Bacillus thuringiensis subspecies israe/ensis Strain BMP 144 (PC Code 

006520); Submitted by B2E Microbials, LLC; MAID 47373303. Memorandum from S. Borges toM. 

Mendelsohn dated October 29, 2008. 

U.S. EPA. 2011 a. BPPD review: Product Identity, Manufacturing Process, Analysis of Samples for Advanced 

Microbiologies' Product Registration (EPA Reg. No. 84268-1). Memorandum by I. Barsoum toM. 

Mendelsohn Word file dated February 1 0, 2011. 

U.S. EPA. 2011 b. BPPD review: Product Chemistry Supplemental to Product Identity, Manufacturing Process, 

Physical and Chemical Characteristics for Advanced Microbiologies' Product Registration (EPA Reg. No. 

84268-1) MAID 47898202. Memorandum by I. Barsoum toM. Mendelsohn Word file dated February 10, 

2011. 

U.S. EPA. 2011c. BPPD review: Product Identity, Analysis of Samples, Physical and Chemical Characteristics, 

Storage Stability MAID 47898301 for Advanced Microbiologies' Product Registration (EPA Reg. No. 

84268-1 ). Memorandum by I. Barsoum toM. Mendelsohn Word file dated February 10, 2011. 

U.S. EPA. 2011d. BPPD review: Acute Dermal Toxicity- rat for Advanced Microbiologies' Product Registration. 

Memorandum by I. Barsoum to M. Mendelsohn Word file dated February 10, 2011. 

U.S. EPA. 2011e. BPPD review: Review of Product Chemistry, Manufacturing Process data, and Acute Toxicities 

studies of the new product AM61432 Primary Powder (EPA Reg. No. 84268-R) containing the active 

ingredient Bacillus sphaericus strain 2362- summary all 84628-1. Memorandum from I. Barsoum toM. 

Mendelsohn Word file dated February 28, 2011. 

U.S. EPA. 2011f. Review of the registrant's response to the data deficiencies found by the Agency in its review of 

product chemistry, manufacturing process of the new product AM61432 Primary Powder containing the 

active ingredient Bacillus sphaericus strain 2362 to demonstrate similarity to ABTS 1743. Memorandum 

from I. Barsoum to M. Mendelsohn dated March 03, 2011. 

U.S. EPA. 2011 g. BPPD review: Product Chemistry and Efficacy Review for FourStar CRG. Memorandum from 

K. Martin to S. Johnson dated September 07, 2011. 

U.S. EPA. 2012. BPPD review: FourStar® MBG containing Bacillus thuringiensis subspecies BMP 144 and 

Bacillus sphaericus 2362, Serotype H5a5b, strain AM614. Memorandum from J. Gagliardi to S. Bacchus 

dated October 03, 2012. 

31