23 

## T

ABLE OF 

## C ONTENTS



### ACRONYMS

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

A. Intr oducti on .............................................................................................................. ............ 6

B. Background Information .................................................................................................... ... 6

1. Use Profile ............................................................................................................... ...... 7 2. Exemption from the Requirement of Tolerance .............................................................. 7 3. Other Regulatory Considerat ions .................................................................................. 8 4. General Manufacturing Proce ss for Microbial Pestici des............................................. 10 5. Biologic Cons iderations ............................................................................................... 10

C. Anticipated Data N eeds and Risk C oncerns ....................................................................... 11
1. Data for Risk Asse ssment............................................................................................. 11 2. Efficacy and Label Cla ims ........................................................................................... 12

3. Endocri ne Effe cts ........................................................................................................ 12

4. In cident s ................................................................................................................. ..... 13

D. Timeline

.............................................................................................................................. 13

E. Guidance fo r Commenters ................................................................................................. 1 4

F. Environment al Just ice ..................................................................................................... .... 14

G. Water Qualit y ............................................................................................................. ........ 14

H. Trade Ir ritants ........................................................................................................... .......... 14

I. Next Steps ................................................................................................................ ........... 14

REFERENCES ........................................................................................................................ 15

APPENDIX I : Pesticide Products Containing B. thuringiensis as an Active Ingredient .. App I-1

APPENDIX II : Summary of Incidents Involving Exposure to B. thuringiensis ................ App II-1

APPENDIX III : History of the To lerance Exemption for B. thuringiensis ........................ App III-14 

### ACRONYMS Acronym Extension

a.i. Active ingredient
ATTC American Type Tissue Collection

B. anthracis Bacillus anthracis

B. cereus Bacillus cereus
B. thuringiensis Bacillus thuringiensis
CFR Code of Federal Regulations

DCI Data Call-in
EDSP Endocrine Disruptor Screening Program
EP End-use product
EPA U.S. Environmental Protection Agency or, the Agency
EPA Reg. No. EPA Registration Number: a two-part number assigned by EPA to identify each pesticide product registration. FFDCA Federal Food, Drug, and Cosmetic Act
FIFRA Federal Insecticide, Fungicide, and Rodenticide Act
FQPA Food Quality Protection Act of 1996
FR Federal Register

FWP Final Work Plan IDS Incident Data System MP Manufacturing-use product
MPCA Microbial pesticide control agent

MRL Maximum residue limits

NPIC National Pesticides Information Center OPP EPA’s Office of Pesticide Programs
OPPIN Office of Pesticide Programs Information Network
PC Code Pesticide Chemical Code: a six-digit number assigned by OPP to identify pesticide chemicals. RED Reregistration Eligibility Determination (RED) TMDL Total maximum daily loads5 

# B ACILLUS THURINGIENSIS 

# R EGISTRATION 

# R EVIEW 

# T EAM



## Office of Pesticide Programs Biopesticides and Pollut ion Prevention Division

Microbial Pesticides Branch

S CIENCE 

## A SSESSMENT



### Kathleen Martin; MSES, MPA Alan Reynolds; MS



## R ISK 

## M ANAGEMENT



### Sheryl Reilly, Ph.D., Chief, Microbial Pesticides Branch



## O FFICE OF 

## G ENERAL 

## C OUNSEL



### Philip Ross, Esq.Bacillus thuringiensis Case 0247 EPA-HQ-OPP-2011-0705 regulations.gov

6 

## P RELIMINARY 

## W ORK 

## P LAN

A. Introduction

The Food Quality Protection Act (FQPA) of 1996 mandated the continuous review of existing pesticides. All pesticides distributed or sold in the United States must generally be registered by the U.S. Environmental Protection Agency (EPA or the Agency), 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. Changes in science, public policy, and pesticide use practices will occur over time. Through the Registration Review program, the Agency 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 at: www.epa.gov/oppsrrd1/registration_review/.

The Agency is implementing the Registration Revi ew 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 Agency will consider benefits information and data, as requi red by FIFRA. The public phase of Registration Review begins when the initial docket is opened for each case. The docket is the Agency’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 comment period, the Agency will develop and commit to a Final Work Plan (FWP) and schedule for this registration review.

This Registration Review Summary Document in cludes the Preliminary Work Plan and provides an initial evaluation of the microbial pest control agent (MPCA) Bacillus thuringiensis . Further information about this case, including additional background and supporting documents, is available at regulations.gov; under “Keyword” enter the doc ket number, which is EPA-HQ-OPP-2011-0705.

B. Background Information

Bacillus thuringiensis is a naturally occurring organism commonly found in soil. It forms parasporal crystalline bodies, which are toxic for certain invertebrates, especially species of insect larvae belonging to the insect orders Coleoptera (beetles), Diptera (flies) and Lepidoptera (butterflies and moths) (Bravo 2007). The toxicity of the crystalline bodies is strain-specific, that is, not all B. thuringiensis strains are toxic to all insects.

B. thuringiensis was first discovered in Japan (1901), by Ishiwata during his study of a bacterial disease of silkworms. Later, in 1915, a German scientist named Berliner found a similar bacillus killing the larvae of flour moths; he published a description of the bacterium and its properties, naming it B. thuringiensis 1 

### . In the 1930’s, French researchers recognized its potential for use in agriculture and developed a product known as Sporeine. (Federici 2005) In 1960, the U.S. Food and Drug Administration established an exemption from the require ment of tolerance for residues of viable spores of the microorganism Bacillus thuringiensis Berliner (25 FR 3207; April 14, 1960); this exemption was republished after EPA was established (36 FR 22572; November 25, 1971). Subsequently, the tolerance exemption has been amended. 2 

### In 1961, B. thuringiensis was registered as a pesticide. Today there are more than 100 registered pesticide products containing B. thuringiensis as an active ingredient. Provided below, under “Regulatory Consider ations,” are details regarding regulatory actions and the tolerance exemption. Appendix I provides a list of all currently registered products containing B. thuringiensis as an active ingredient.

1 The species name “thuringiensis” is derived from Thuringia, t he region in Germany in which the disease occurred (Federici 2005)

. 2 See Appendix III for details regarding the amendments.Bacillus thuringiensis Case 0247 EPA-HQ-OPP-2011-0705 regulations.gov

7 

### 1. Use Profile

P ESTICIDE 

### T YPE 

### : Insecticide

U SE 

### S ITES 

### : Field-, greenhouse-, and water-grown 3 

### crops (pre- and postharvest use)
Food and nonfood storage containers, buildings, and transport vehicles
Outdoor and greenhouse-grown ornamentals and nursery crops
Seed treatment (food and nonfood seeds) Standing water (e.g., certain crop fields, flooded fields, irrigation ditches, livestock watering containers, ornamental ponds, pastures, snowmelt)

R EGISTERED FOR 

### U SE 

### A GAINST


### T HESE 

### P ESTS 

### : Coleoptera (beetles, bugs, weevils)
Diptera (flies, gnats, midges, mosquitoes)
Lepidoptera (borers, butterflies, casebearers, cutworm, leaf folders, leaf perforator, leafrollers, leafworms, loopers, moths, orange dog, skippers)

NOTE: B. thuringiensis products are registered for use against mature insects and insects in one of their various life stages (e.g., 3 rd
instar, egg, larval, pupal).



### F ORMULATION


### T YPES 

### : aqueous suspension, bait, dry flowable, dust, emulsifiable concentrate, emulsifiable suspension, granule, slurry, technical powder, water dispersible granule, water soluble pouch, wettable powder

A PPLICATION


### M ETHODS 

### : Aerial (fixed wing and rotary aircraft), low- and ultra-low volume application (aqueous or oil spray), tank mix, hand sprayer

B ASIC


### M ANUFACTURERS 

### : Becker Microbial Products, Inc. (Company Number: 62637)
Certis USA, LLC (Company Number: 70051) Valent Biosciences Corporation (Company Number: 73049)

2. Exemption from the Requirement of Tolerance

The current version of the exemption reads as follows:

§ 180.1011 Viable spores of the microorganism Bacillus thuringiensis Berliner; exemption from the requirement of a tolerance.

(a) For the purposes of this section t he microbial insecticide for which exemption from the requirement of a tolerance is being established shall have the following specifications:

(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 wi th adequate control measures during production to detect any changes from the ch aracteristics of the parent strain or contamination by other microorganisms.

3 Water-grown crops include cranberry bogs and rice fields.Bacillus thuringiensis Case 0247 EPA-HQ-OPP-2011-0705 regulations.gov

8 (3) Each lot of spore preparation, prior to the addition of other materials, shall be tested by subcutaneous inject ion 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 te st 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 (“Mic robial Control of Insects and Mites,” R.P.M. Bond et al., p. 280 ff., 1971). This specificati on 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.

(b) Exemption from the requirement of a tolerance is established for residues of

the microbial insecticide Bacillus thuringiensis Berliner, as specified in paragraph (a) of this section, in or on honey and hon eycomb 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.

[36 FR 22540, Nov. 25, 1971, as amended at 38 FR 19045, July 17, 1973; 42 FR 28540, June 3,1977; 45 FR 43721, June 30, 1980; 45 FR 56347, Aug. 25, 1980; 74 FR 26533, June 3, 2009]



### 3. Other Regulatory Considerations

Reregistration . The 1988 amendments to FIFRA authorized the Agency to conduct a comprehensive pesticide reregistration program—a complete reassessment of the human health and environmental effects of pesticides first registered before November 1, 1984— to make decisions about these pesticides' future sale, distribution, and use. 4 

### The results of the reassessments were later summarized in Reregistration Eligibility Decision (RED) documents. A Registration Standard for B. thuringiensis was published in 1988 (U.S. EPA 1988) and the RED for the reregistration review of B. thuringiensis was issued in 1998 (U.S. EPA 1998). The

RED concluded that B. thuringiensis products were eligible for reregistration; it pointed out that B. thuringiensis may produce undesirable toxins (see “Biologic Considerations,” below, for a discussion on the undesirable toxins). The RED also stated the following: “To mitigate risks of potential toxicity to the public and/or non target species from these toxins, the Agency is requiring continuation of the production batch quality control testing that originally appeared in the tolerance exemption and is requiring t he reevaluation and standar[d]ization of the manufacturing process for each registered technical grade of the active ingredient” (U.S. EPA 1998). This RED element is also addressed in the tolerance exemption and in 40 CFR 158.

Registered Products . Pesticide products containing B. thuringiensis as an active ingredient are of one of four subspecies:

 B. thuringiensis subspecies israelensis ;

· B. thuringiensis subspecies kurstaki ;

· B. thuringiensis subspecies aizawai ; or

· B. thuringiensis subspecies tenebrionis

.
Each subspecies, in turn, has one or more strains of B. thuringiensis . Table 1 is a summary of Appendix I—it shows the subspecies and their stra ins but only provides how many active pesticide products are in each strain. Three of the entries (PC Codes 006401, 006402, and 006405) show end-use products (EPs) or FIFRA section 24(c) registrations but no analogous manufacturing-use product (MP). PC Codes 006401, 006402, and 006405 are the generic
PC Codes for B. thuringiensis subspecies israelenis

, kurstaki , and tenebrionis, respectively.

4 www.epa.gov/oppsrrd1/reregistration/reregistration_facts.htm.Bacillus thuringiensis Case 0247 EPA-HQ-OPP-2011-0705 regulations.gov

9 

### At one time all B. thuringiensis i sraelensis

, kurstaki , and tenebrionis registrations, regardless of strain, were listed in the Agency’s Office of Pesticide Programs Information Network (OPPIN) database under the generic PC Code. Later, the Agency began assigning individual PC Codes for each strain. During the course of registration review, EPA will correct this administrative artifact by assigning the strain-specific PC Codes to these products. Another curiosity is with B. thuringiensis kurstaki strain EG7841 Lepidopteran active toxin, PC Code 006453—there is a single entry for two products, and the number of EPs versus MPs is not differentiated. The reason for this is that these two products are made by an integrated process, where the EP is made from start to finish in a single stream—a separate MP is not formed. This is a common practice with some microbial pesticides. More information on the manufacturing process is provided below, under “General Manufacturing Process of Microbial Pesticides.”

Table 1

. Bacillus Thuringiensis Subspecies and Strains

S TRAIN PC
C ODE N O

. A CTIVE P RODUCTS N OTES MP EP 24( C )

B. thuringiensis subspecies israelensis

B. thuringiensis israelensis 006401 -- 4 1 During this registration re view, EPA will reclassify these five products—PC Code 006401 is the generic category for B. thuringiensis israelensis ; these five products should be coded by their particular strains.

B. thuringiensis israelensis
strain EG2215 006476 1 -- --

B. thuringiensis israelensis strain BMP 144 solids, spores and insecticidal

toxins 006520 1 14 --

B. thuringiensis israelensis
AM 65-52 069162 3 13 1 Two of the MPs are solids; one is a slurry.

B. thuringiensis israelensis
strain SA3A 069210 2 3 -- One MP is a solid and the other is a slurry.

B. thuringiensis subspecies kurstak i

B. thuringiensis kurstaki 006402 -- 3 2 During this registration re view, EPA will reclassify these five products—PC Code 006402 is the generic category for B. thuringiensis kurstaki ; these five products should be coded by their particular strains.

B. thuringiensis kurstaki

strain BMP123 006407 1 4 --

B. thuringiensis kurstaki
strain EG2371 006423 1 1 --

B. thuringiensis kurstaki
strain EG2348 006424 1 3 --

B. thuringiensis kurstaki strain EG7841 Lepidopteran

active toxin 006453 2 -- Integrated Process (i.e., there is no technical or MP)

B. thuringiensis kurstaki strain EG7826 Lepidopteran

active toxin 006459 1 1 --

B. thuringiensis kurstaki strain SA-12 solids, spores, and insecticidal toxins,
ATCC # SD-1323 006518 2 12 -- Both the MPs are solids.Bacillus thuringiensis Case 0247 EPA-HQ-OPP-2011-0705 regulations.gov

10 S TRAIN PC
C ODE N O

. A CTIVE P RODUCTS N OTES MP EP 24( C )

B. thuringiensis kurstaki strain SA-11 solids, spores and insecticidal toxins,
ATCC # SD-1322 006519 1 4 --

B. thuringiensis kurstaki
strain ABTS-351 006522 4 29 1 Two of the MPs are solids and two are slurries.

B. thuringiensis subspecies aizawa i

B. thuringiensis aizawai
strain GC-91 006426 1 2 --

B. thuringiensis aizawai
strain NB200 006494 1 -- --

B. thuringiensis aizawai
strain ABTS 1857 006523 3 3 -- Two of the MPs are solids; one is a slurry.

B. thuringiensis subspecies tenebrionis

B. thuringiensis tenebrionis 006405 -- -- 1

B. thuringiensis tenebrionis

strain NB-176 006524 1 2 --



### 4. General Manufacturing Process for Microbial Pesticides (Taborsky 1992)

Microbial pesticides are made by fermentat ion, where a starter culture is added to a broth of nutrients so that the bacteria can grow. The end result is a slurry, which contains unused broth, fermentation solids, spores, and insecticidal toxins. This slurry can then be dried down to a solid by centrifugation and drying in air or an oven. For some microbial products the slurry is considered to be the manufacturing-use product (MP). For others, the MP is the dry form, where the excess moisture has been removed. After fermentation but before drying, inert ingredients such as wetting agents may be added. Some end-use products are made without an intermediate MP, as the end-use product (EP) is made from start to finish in a single ‘integrated process.’ Crymax (EPA Reg. No. 70051-86) and Crymax WP (EPA Reg. No. 70051-90) are two B. thuringiensis -containing products that are made this way. See Appendix I for the complete list of the B. thuringiensis -containing products.

5. Biologic Considerations

The scientific classification for B. thuringiensis is (NIH 2011):

K INGDOM 

### : Bacteria

P HYLUM 

### : Firmicutes

C LASS 

### : Bacilli

O RDER 

### : Bacillales

F AMILY 

### : Bacillaceae

G ENUS 

### : Bacillus

S PECIES 

### G ROUP 

### : Bacillus cereus

S PECIES 

### : thuringiensBacillus thuringiensis Case 0247 EPA-HQ-OPP-2011-0705 regulations.gov

11 

### Members of the genus Bacillus are gram-positive, aerobic (or facultative anaerobic), endospore-forming bacteria. It is this ability to form spores that allows these bacteria to survive in soil. (Todar 2011). B. thuringiensis is one of three species of the Bacillus cereus group (the others being B. cereus and Bacillus anthracis ) that have an impact on human activity (Vilas-Bôas 2007). B. cereus and B. anthracis are human pathogens. B. cereus causes two forms of foodborne illness via production of exotoxin s. The emetic form, which is characterized by nausea and vomiting, is caused by emetic toxin and the more severe diarrheal form, which manifests as abdominal cramps and diarrhea, is caused by nonhemolytic enterotoxin and/or hemolytic enterotoxin. B.anthracis causes anthrax, which is primarily a disease of domesticated and wild animals; humans may become infected when brought into contact with diseased animals. In the United States, the incidence of naturally-acquired human anthrax is extremely rare (one to two cases of cutaneous disease per year) (Todar 2011).

B. thuringiensis is geno- and phenotypically related to B. cereus and B.anthracis . It is genetically indistinguishable from B. cereus , except for the ability of B. thuringiensis to produce parasporal crystalline bodies, which are toxic for certain invertebrates, especially species of insect larvae belonging to the insect orders Coleoptera

, Diptera and Lepidoptera

. The crystals are predominantly comprised of one or more proteins (Cry and Cyt toxins), which are also called

δ -endotoxins. They have various shapes (e.g., bipyramidal, cuboidal, flat rhomboid, spherical) depending on their protein composition. Co rrelation between crystal morphology, protein composition, and bioactivity against target insects has been established. (Bravo 2007; IPCS 1999)

C. Anticipated Data Needs and Risk Concerns

1. Data for Risk Assessment

B. thuringiensis is a complex registration review case—not only are there a great number of strains, each with its own data set for the three scientific disciplines (i.e., product analysis, toxicology, and nontarget organisms and environmental fate), as set forth in 40 CFR 158, subpart V, but the Agency’s internal system for tracking data and data reviews has changed over time. EPA uses PC Codes 5 

### to track and file data submitted to support registration and the Agency’s data reviews. At one time, the same PC Code was used for all strains of a particular subspecies; for example, all strains of B. thuringiensis israelensis were coded under PC Code 006401. In 1989, EPA started assigni ng individual PC Codes for each new strain. Some of the data and reviews are still coded under the old system, which has presented challenges thus far in the registration review of B. thuringiensis

.
EPA has completed a preliminary review of the B. thuringiensis database and has identified some data gaps. For example, the Product Analysis requirement for ‘deposition of a sample in a nationally recognized culture collection’ (40 CFR 158.2120) has not been satisfied for a number of strains. This can be expected, as many of the B. thuringiensis -containing products were registered before the October 26, 2007 Final Rule; this rule, among other things, revised the microbial data requirements to include the aforementioned data requirement for culture collection deposition (72 FR 60988; October 26, 2007). As the Agency develops its FWP for this case, it will also complete a comprehensive evaluation of its entire record for B. thuringiensis . To date, EPA has only reviewed the information available in its electronic databases. The Agency is now in the process of examining the paper files for B. thuringiensis

, which includes product jackets housed in the Federal Record Center. In the FWP, a full accounting of how each data requirement has (or has not) been met will be provided. Also in the FWP, EPA will discuss its findings and will finalize its conclusion s regarding the need for
additional data for risk assessment.

5 A PC Code (Pesticide Chemical Code) is a six-digit number assigned by OPP to identify and track pesticide chemicals, inert ingredients, and other chemicals in its internal databases.Bacillus thuringiensis Case 0247 EPA-HQ-OPP-2011-0705 regulations.gov

12 

### In addition to the comprehensive review of its entire record for B. thuringiensis , EPA plans to complete the following during the registration review of B. thuringiensis :

· Review the tolerance exemption to determine if it needs to be updated or amended to

take into account advances in microbial analytic techniques and microbiology in general. Also, the current exemption is broad, it only considers B. thuringiensis at the genus and species level; EPA intends to investigate whether the exemption should be expanded to consider lower taxonomic levels such as subspecies and strain.

· Reassess B. thuringiensis ’ hazard to threatened and endangered species, and review

current ecological risk mitigation measures.

· Review the science data sets for each strain of the technical products to make sure that they are current with complete bibliographies.

2. Efficacy and Label Claims

The Agency requires and reviews efficacy data for microbial pesticides making labeling claims to control public health pests. Public health pests are either: (1) pest microorganisms infectious to man in any area of the inanimate environment; or (2) vertebrates or invertebrates (including but not limited mosquitoes and ticks) that may directly or indirectly transmit diseases to humans (40 CFR 158.2160). Because B. thuringiensis is registered for use against mosquitoes, efficacy data are required. As part of its comprehensive review of the entire B. thuringiensis record, the Agency will ensure that all efficacy data requirements have been met. If efficacy data gaps are identified, EPA will discuss them in the FWP.

3. 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 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 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 contains 58 pesticide active ingredients and nine inert ingredients. B. thuringiensis is not among the group of 58 pesticide active ingredients on the
initial list to be screened under the EDSP.

The Agency, as part of this preliminary work plan, believes that B. thuringiensis , the active ingredient involved in this registration review case, 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, pursuant to Section 408(p)(4), EPA will determine in the future whether it can exempt B. thuringiensis from the requirements of the Section 408(p) EDSP. In the event the 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: www.epa.gov/endo/.Bacillus thuringiensis Case 0247 EPA-HQ-OPP-2011-0705 regulations.gov

13 

### 4. Incidents

The National Pesticides Information Center (NPIC) database reported no incidents associated with products containing B. thuringiensis . The Office of Pesticide Programs’ (OPP) Incident Data System (IDS) shows 40 human or domestic animal reports of adverse effects following exposure to a pesticide product containing B. thuringiensis . Twenty-nine are human cases that are considered to be minor; many involved being sprayed accidentally during aerial application and a few were attempted suicides. Eleven cases involved domestic animals. In a few of the cases the animals died or had to be euthanized. In others they experienced mild to moderate effects. For example, in one case a Great Dane had diarrhea for two days. For more information on the incidents, see Appendix III. The Agency will consider any incident data or comments submitted to this docket in response to this preliminary work plan.

D. Timeline

The projected timeline for case 0247—The Bacillus thuringiensis Registration Review— is shown in Table 2. Please note that this schedule is subject to revision should unforeseen issues arise during the registration review process. In the event an issue arises, such as the failure to acquire an EDSP exemption, an amended FWP will be issued. As also discussed earlier, the FWP will identify any newly anticipated data needs and set forth a new timeline if necessary.

Table 2. Projected Timeline for the Registration Review of Case 0247

A CTIVITY E STIMATED M ONTH AND Y EAR

Phase 1 : Open the Public Docket

Open Comment Period for Docket September 30, 2011

Close Comment Period November 30, 2011

Phase 2 : Case Development

Develop Final Work Plan May 2012

Issue DCI May 2013

Data Submission May 2015

Open Public Comment Period for Preliminary Risk Assessments November 2016

Close Public Comment Period December 2016

Phase 3 : Registration Review Decision

Open Comment Period for Proposed Registration Review Decision June 2018

Close Comment Period August 2018

Final Decision and Begin Post-Decision Followup 2018

Estimated Total (years) 7Bacillus thuringiensis Case 0247 EPA-HQ-OPP-2011-0705 regulations.gov

14 

### E. Guidance for Commenters

The public is invited to comment on EPA’s preliminary work plan and rationale. The Agency will consider all comments as well as any additional inform ation or data provided in a timely manner prior to issuing an FWP for the case.

F. Environmental Justice

EPA seeks to achieve environmental justice, t he 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, regulati ons, and policies. To help address potential environmental justice issues, the Agency 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 B. thuringiensis , 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.

G. Water Quality

B. thuringiensis is not identified as a cause of impairment for any water bodies listed as impaired under section 303(d) of the Clean Water Act. 6 

### In addition, no Total Maximum Daily Loads (TMDL) have been developed for B. thuringiensis

. 7 

### More information on impaired water bodies and TMDLs can be found at: www.epa.gov/owow/tmdl/. The Agency invites submission of water quality data for this pesticide. To the extent possible, data should conform to the quality standards in Appendix A of OPP Standard Operating Procedure: Inclusi on of Impaired Water Body and Other Water Quality Data in OPP’s Registration Review Risk Assessment and Management Process 8 

### to ensure they can be used quantitatively or qualitatively in pesticide risk assessments.

H. Trade Irritants

Through the registration review process, the Agency intends to solicit information on trade irritants and, to the extent feasible, take steps toward facilitating irritant resolution. Growers and other stakeholders are asked to comment on any trade irritant issues resulting from lack of Maximum Residue Limits (MRLs) or disparities between United States tolerances and MRLs in key export markets, providing as much specificity as possible regarding the nature of the concern.

Residues of B. thuringiensis are exempt from the requirement of tolerance and there are no known MRLs (FAO and WHO 2010). Therefore, the Agency does not anticipate that current uses of B. thuringiensis will pose concerns as trade irritants.

I. Next Steps

After the 60-day comment period closes, the Agency will review and respond to any comments received in a timely manner and then issue an FWP for Bacillus thuringiensis case 0247.

6 Based on information provided at: http://www.epa.gov/owow/tmdl/.

7 Based on information provided at: http://iaspub.e pa.gov/tmdl_waters10/text_search.tmdl_search_form.

8 This document can be found at: http://www.epa.gov/ oppsrrd1/registration_review/water_quality_sop.htm.Bacillus thuringiensis Case 0247 EPA-HQ-OPP-2011-0705 regulations.gov

15 

### REFERENCES

Bergey. 2009. Bergey’s Manual of Systematic Bact eriology, Volume 3; 2nd Ed. Springer. New York.

Bravo, A. 2007. Mode of action of Bacillus thuringiensis Cry and Cyt toxins and their potential for insect control. Toxicon. March 15; 49(4):423–435.

FAO and WHO. 2010. Codex Alimentarius. United Nations Food and Agriculture Organization & World Health Organization. Information retrieved on June 9, 2011. www.codexalimentarius.net/mrls/pestdes/jsp/pest_q-e.jsp

Federici, BA. 2005. Insecticidal bacteria: An overwhelming success for invertebrate pathology. Journal of Invertebrate Pathology 89 (2005) 30–38.

IPCS. 1999. Environmental Health Criteria 217: Bacillus thuringiensis. International Programme on Chemical Safety. United Nations Environment Pr ogramme and World Health Organization. Geneva. www.inchem.org/pages/ehc.html

NIH 2011. Taxonomy Browser. U.S. Department of Health and Human Services. National Institutes of Health. National Library of Medicine. National Center for Biotechnology Information. Information retrieved on September 15, 2011. www.ncbi .nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi

Taborsky, V. 1992. Small-Scale Processing of Microbial Pesticides. FAO Agricultural Services Bulletin No. 96. Food and Agriculture Organization of the United Nations. Rome. www.fao.org/docrep/t0533E/t0533E00.htm.

Todar, Kenneth. 2011. Todar’s Online Textbook of Bacter iology. University of Wisconsin. Information retrieved on September 15, 2011. www.textbookofbacteriology.net

U.S. EPA. 1988. Guidance for the Reregistration of Pesticide Products Containing Bacillus thuringiensis as the Active Ingredient. U.S. Envir onmental Protection Agency. Office of Pesticides and
Toxic Substances. December 1988. 540/RS-89-023.

U.S. EPA. 1998. Reregistration Eligibility Decision (R ED): Bacillus thuringiensis. U.S. Environmental Protection Agency. Office of Prevention, Pest icides, and Toxic Substances. March 1998. EPA738-R98-004.

Vilas-Bôas, G.T., et. al. 2007. Biology and taxonomy of Bacillus cereus

, Bacillus anthracis , and Bacillus thuringiensis . Can. J. Microbiol. 53:673-687.APP I-1 

### A PPENDIX 

### I : P ESTICIDE 

### P RODUCTS 

### C ONTAINING 

### B

ACILLUS THURINGIENSIS AS AN 

### A CTIVE 

### I NGREDIENT 

### (A.I.)

Provided below are the currently registered pesticide products containing Bacillus thuringiensis as an active ingredient; they include section (3) and 24(c) registrations. Highlighted entries denote manufacturing-use products (MP).

S UBSPECIES

, S TRAIN

AND PC C ODE EPA

R EG

. N O

. N AME % A.I. N OTES

Bacillus thuringiensis subspecies israelensis

006401 B. thuringiensis

israelensis 4-455 Bonide Mosquito Beater WSP 1.7
6218-73 Summit Mosquito Bits 2.86

70051-102 Bti Granule Larvicide 2.86

81045-1 Healthy Outdoors Brand Sustained Release Mosquito Larvicide 15

AZ960003 Vectobac G Biologica l Larvicide Granules 0.2 Parent sec(3) registration is 73049-10

006476 B. thuringiensis israelensis strain
EG2215 73049-405 Bti Technical Powder Bioinsecticide 100 MP

006520
B. thuringiensis israelensis

strain BMP 144 solids, spores and

insecticidal toxins 62637-7 BMP 144 Primary Powder 100 MP

432-1515 BES0531 10
769-992 Allpro Sustain BCG 5.71
6218-47 Summit B.T.I. Briquets 10.31
62637-1 BMP 144 (2x) 8
62637-2 BMP 144 (3x) 12
62637-3 BMP 144 (200 G) 2.86

62637-11 BMP 144 DF 17.15

62637-12 BMP 144 DFX 25.7
62637-13 BMP 144 (400 G) 5.71

62637-14 BMP 144DF 3000 43

83362-2 Meridian-02 7
83362-3 FourstarTM Briquets 1 a

85685-1 Fourstar TM SBG 2.15

85685-2 Fourstar TM CRG 1 a

069162
B. thuringiensis israelensis

strain AM 65-52 73049-13 Vectobac Technical Powder 99 MP

73049-355 Vectobac Biological Larvicide Primary Powder 99 MP

73049-61 Bacillus Thuringiensis subspecies Israelensis Slurry 14 MP

8329-39 Bti Granules 0.25

9688-212 Chemsico Insect Granules Ml 4.95
73049-10 Vectobac G Biological Larvicide Granules 2.8

73049-11 Vectobac- AS 6.38

73049-18 Vectobac-12 AS-II 11.6

73049-19 Vectobac GR 4.95
73049-38 Vectobac 12AS Biological Larvicide 11.6
73049-56 Vectobac WDG Biological Larvicide 37.4

73049-429 V60035 CG 4.5 a

73049-447 Vectobac DT Biological Larvicide (VBC-60046) 37.4
73049-451 VBC-60092 (Bactimos Briquette) 10.3
73049-452 Vectobac Pellet Biological Larvicide 12.3
73049-477 Bacillus thuringiensis subs pecies Israelensis Slurry 14.0

DE090002 Vectobac WDG 37.4 Parent sec(3) registration is 73049-56

069210
B. thuringiensis israelensis
strain SA3A 73049-408 Teknar Technical Concentrate 100 MP

73049-428 Teknar Slurry 22 MP

73049-403 Teknar G 1.7APP I-2 S UBSPECIES

, S TRAIN

AND PC C ODE EPA

R EG

. N O

. N AME % A.I. N OTES

73049-404 Teknar HP-D 1.6
73049-435 Teknar HP Biological Larvicide Aqueous Suspension 5.6

a Contains Bacillus sphaericus as a second active ingredient (83362-3, 6%; 85685-2, 9%; and 73049-429, 2.7%).

Bacillus thuringiensis subspecies kurstak i

006402
B. thuringiensis kurstaki 73049-406 Thuricide 76 LV 14.4
73049-413 Thuricide 48LV Forestry 19

85827-3 Green Light Bt Worm Killer 15
ND010004 Dipel ES 23.7 Parent section (3) registration is 73049-17 TX020001 Dipel ES 23.7

006407
B. thuringiensis kurstaki

strain BMP123 62637-9 BMP 123 Primary Powder 100 MP
62637-5 BMP 123 (2X WP) 40
62637-6 BMP 123 (48 LC) 2.15
62637-8 BMP 123 (10G) 2
62637-10 BMP 123 (64ES) 22

006423
B. thuringiensis kurstaki

strain EG2371 70051-84 Cutlass Technical Powder 100 MP

70051-79 Cutlass 40

006424
B. thuringiensis

kurstaki strain EG2348 70051-83 Condor Technical Powder 100 MP

70051-78 Condor 24.5

70051-80 Condor WP 43

70051-85 Condor XL 45

006453
B. thuringiensis kurstaki strain EG7841Lepidopteran active toxin 70051-86 Crymax 40 Integrated process a 70051-90 Crymax WP 40

006459
B. thuringiensis kurstaki strain EG7826 Lepidopteran active toxin 70051-87 Lepinox 100 MP

70051-89 Lepinox WDG 40

006518
B. thuringiensis kurstaki
strain SA-12 solids, spores, and insecticidal toxins,

ATCC # SD-1323 70051-68 Thuricide Technical Concentrate 100 MP

70051-70 San 420 I ® Technical Concentrate 100 MP

4-226 Bonide Bacillus thuringiensis (Bt) Moth Larvae (Caterpillar) 15

4-487 Bonide Bacillus Thuringiensis "O" 98.35

829-202 SA-50 Brand Thuricide HPC 15
2724-688 Security Bt Dust Biological Insecticide 0.049

9688-294 Chemisco HPC-O Concentrate 98.35

70051-52 Thuricide HPC 15

70051-53 Thuricide HP WP 70

70051-57 Thuricide 48 LV 19

70051-65 Thuricide HPC 15

70051-69 San 420 I WG 85

70051-103 Btk32 85

70051-106 Thuricide HPC-O For Home
And Gardens 98.35

006519
B. thuringiensis kurstaki strain SA-11 solids, spores and insecticidal toxins,
ATCC # SD-1322 70051-67 Javelin Technical Concentrate 100 MP

70051-60 Javelin 17.8

70051-61 Javelin WP 80

70051-66 Javelin WG 85

70051-98 Javelin WG 2 85

006522
B. thuringiensis kurstaki
strain ABTS-351 73049-12 Dipel Technical Powder 91 MP

73049-27 ABG-6345 Technical Powder 84 MP

73049-62 Bacillus thuringiensis subspecies Kurstaki Slurry 22 MP

73049-476 Bacillus thuringiensis subspecies Kurstaki Slurry (for Foray Products) 22 MP

4-251 Bonide Dipel 150 Dust For Vegetables 0.47
4-252 Dipel WP Home & Garden Insecticide 6.4APP I-3 S UBSPECIES

, S TRAIN

AND PC C ODE EPA

R EG

. N O

. N AME % A.I. N OTES

4-336 Dipel 10G Sweet Corn Granules 2.3
4-405 Dipel Bio Garden Spray 6.4
829-196 SA-50 Brand Dipel Dust 0.41

2935-398 Wilbur-Ellis Bt 320 Dust 0.582

2935-399 Bt 320 Sulfur 25 Dust 0.582 a

7401-290 Ferti-Lome Dipel Dust Bi ological Insecticide 0.064

10951-10 Britz Bt 25 Sulfur Dust 1.16

10951-17 Britz Bt Dust 1.94

34704-722 Bt Sulfur 15-50 Dust 0.72
36488-25 Ringer Vegetable Insect Attack 0.4365
42697-23 Safer B.T. Caterpillar Killer Concentrate 12.74

73049-5 Dipel (Worm Killer) Wettable Powder Biological Insecticide 23.7

73049-8 Dipel 2x Biological Insecticide 58.2
73049-14 Dipel 10G Biological Insecticide Granules 2.3
73049-17 Dipel ES 23.7

73049-30 Dipel ES-NT Biological Insecticide Emulsifiable Suspension 25

73049-33 Dipel SG Plus Biological Insecticide Sand Granule 2.5
73049-34 Dipel WDG Biological Insecticide 63.5
73049-35 Foray 48F Biological Insecticide Flowable Concentrate 17.6
73049-39 Dipel DF Dry Flowable Biological Insecticide 54
73049-46 Foray 48BC 17.19
73049-49 Foray 76 B 18.44
73049-50 Biobit HP WP II Biological Insecticide 34.11
73049-54 Biobit HP Biological Insecticide Wettable Powder 58.2
73049-427 Foray 48B 57.5
73049-471 Foray WG 76.2

85827-4 Green Light Dipel Dust 0.49

CA100012 Dipel DF Biological Insecticide 54 Parent section (3) registration is 73049-39

a These products contain sulfur as a second active ingredient (2935-399 and 10951-10 at 25% and 34704-722 at 50%).

Bacillus thuringiensis subspecies aizawa i

006426
B. thuringiensis aizawai
strain GC-91 70051-48 Technical CGA-237218 100 MP
70051-47 Agree 50
70051-99 Jackpot WP 50

006494
B. thuringiensis aizawai

strain NB200 73049-68 Florbac Slurry Biological Larvicide 18 MP

006523
B. thuringiensis aizawai
strain ABTS 1857 73049-24 Xentari Biological Insecticide Technical Powder 89 MP

73049-28 ABG-6346 Biological Insecticide 83 MP

73049-63 Bacillus thuringiensis subspecies Aizawai Slurry 18 MP

73049-23 Xentari Biological Insecticide Water Dispersible Granule 48.1
73049-31 Xentari As Biological Insecticide 10.8
73049-40 Xentari Biological Insecticide Dry Flowable 54

Bacillus thuringiensis subspecies tenebrionis

006405
B. thuringiensis tenebrionis WA040029 Novodor Biological Insecticide Flowable Concentrate 10 Parent section (3) registration is 73049-48

006524
B. thuringiensis tenebrionis

strain NB-176 73049-47 Novodor Technical 92.6 MP

73049-64 Novodor Flowable Concentrate 10
73049-48 Bacillus thuringiensis su bspecies Tenebrionis Slurry 17APP II-1 

### A PPENDIX 

### II : S UMMARY OF 

### I NCIDENTS INVOLVING EXPOSURE TO 

### B ACILLUS THURINGIENSIS



### Provided in the following table is a summary, arrayed by subspecies, of incidents compiled in the Agency’s Incident Data System database between 1992 and 2010. Where information was available, discussion regarding the nature of the incident is given.

T OTAL N UMBER OF I NCIDENTS : D ISCUSSION N O

. H UMAN N O

. D OMESTIC A NIMAL

Bacillus thuringiensis subspecies israelensis

11 3=H-D a 3=D-C b

5=D-D c Human Cases
Allergic reaction was reported among farmworkers who were exposed to Bacillus thuringiensis . They exhibited a significant incr ease in positive skin tests to spore

extracts.

Bacillus thuringiensis subspecies kurstak i

26 23=H-D 2=D-A d

2=D-D c Human Cases
A number of the cases involved individuals being sprayed accidentally. In one incident of aerial application, a female experienced an allergic reaction (burning, itching eyes, red skin rash). In another incident a female was sprayed while horseback riding; she experienced eye and lymph node problems, skin irritation, and headache. A man reported being sprayed in the face by a helicopter; he later fell from the roof. Sunbathers who were sprayed by a helicopter developed laryngitis. In a case wher e the Forest Service sprayed an 800 acre housing area, residents developed watery eyes and breathing problems.

Other cases involved incidental exposure after the pesticide was applied. In an

incident where a house was treated for tw o days, the resident had an allergic reaction, most likely due to an inert ingredient. Symptoms included difficultly breathing and plugged ears. A 78 year-old man developed full-body edema one hour after pecan trees were sprayed; a rash appeared five days later. Another case involved five passengers and two cargo handlers of an airplane who became ill with nausea and dizziness; treated bananas were in the cargo.

Several cases involved suicide attempts.

Animal
In a case of aerial spraying, a Great Dane developed diarrhea for two days.

Bacillus thuringiensis subspecies aizawa i

2 H-D Human Cases
California farmworkers experienced allergic reaction after exposure to Bacillus thuringiensis ; they showed significant positive skin tests to spore extracts.

Bacillus thuringiensis subspecies tenebrionis

1 H-D

a H-D is considered to be a minor human inciden t. An incident is classified as H-D if “If the person alleged or exhibited some s ymptoms, but they were minimally traumatic. The symptoms resolved rapidly and usua lly involve skin, eye or respiratory irritation” 40 CFR 159.184 (c)(5).

b D-C is a moderate domestic animal incident. An incident is classified as D-C if “ the domestic animal exhibited or was alleged to have exhibited symptoms which are more pronounced, more prolonged or of a more systemic nature than minor symptoms. Usually some form of trea tment would have been indicated to treat the animal. Symptoms were not life threatening and the animal has returned to its pre-exposure st ate of health with no

additional residual disability” 40 CFR 159.184(c)(5).

c D-D is a minor domestic animal incident. An incident is clas sified as D-D if “the domestic animal was alleged to have exhibite d symptoms, but they were minimally bothersome. The symptoms resolved rapidly and usually involve skin, eye or respirator irritation” 40 CFR 159.18 4(c)(5).

d D-A is a domestic animal death. This code is used if the If the domestic animal died or was euthanized” 40 CFR 159.184(c)(5).APP III-1 

### A PPENDIX 

### III : H ISTORY OF THE 

### T OLERANCE 

### E XEMPTION FOR 

### B ACILLUS THURINGIENSIS



### Provided below is the original tolerance exemption (by the Food and Drug Administration) followed by it’s subsequent amendments and the reasons for the amendments.

25 FR 3207 April 14, 1960 § 120.176 Exemption from the requirement of a tolerance for residues of viable spores of the microorganism Bacillus thuringiensis Berliner.

(a) For the purposes of this section the microbial pesticide for which exemption from the requir ement of a tolerance is being established shall have the following specifications:

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

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 characterist ics 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 subcut aneous 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.

(b) Exemption from the requ irement of a tolerance is established for residues of the microbial pesticide Bacillus thuringiensis Berliner as specified in paragraph (a) of this section in or on the following raw agricultural commodities: Alfalfa, apples, artichokes, beans, broccoli, cabbage, cauliflower, celery, cottonseed, lettuce, potatoes, spinach. Original Tolerance Exemption

38 FR 9045 July 17, 1973 § 180.1011 Exemption from the requirement of a tolerance for residues of viable spores of the Bacillus thuringiensis Berliner; exemption from the requirement of a tolerance.

(a) * * *

(b) 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. EPA amended the ex emption “to include a toxicity test to determine the presence of β -exotoxin to insure that B. thuringiensis products are free of

this toxin.”

42 FR 28540 June 3, 1977 § 180.1011 V iable spores of the microorganism Bacillus thuringiensis, Berliner; exemption from the requirement of a

tolerance.

(a) * * *

(b) Exemption from the requ irement of a tolerance is
established for residues of the microbial insecticide Bacillus thuringiensis, Berliner, as specified in paragraph (a) of this section, in or on all raw agricultural commodities when it is applied either to growing crops or when it is applied after harvest in accordance with good agricultural practice. At the request of IR-4 (Interregional Research Project No. 4), the Agency amended § 180.1011 to exempt
residues of “ Bacillus thuringiensis, Berliner from the requirement of tolerance in or on all raw agricultural commodities when the-pesticide is applied to growing crops.”APP III-2 45 FR 43721 June 30, 1980 § 180.1011 V iable spores of the microorganism Bacillus thuringiensis Berliner , exemption from the requirement of a

tolerance

. (a) * * *

(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. EPA made an amendment to update the Bergey reference from the Seventh edition to the Eighth edition.

45 FR 56347 Aug. 25, 1980 § 180.1011 Viable spores of the microorganism Bacillus thuringiensis Berliner , exemption from the requirement of a

tolerance.



### * * * * *

(b) Exemption from the requ irement of a tolerance is
established for residues of the microbial insecticide Bacillus thuringiensis Berliner, as specified in paragraph (a) of this section, in or on beeswax and honey 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. At the request of IR-4 (Interregional Research Project No. 4), the Agency amended § 180.1011 to exempt
residues of “ Bacillus thuringiensis Berliner in or on the raw agricultural commodities beeswax and honey.”

74 FR 26533 June 3, 2009 § 180.1011 Viable spores of the microorganism Bacillus thuringiensis Berliner; exemption from the requirement of a

tolerance.



### * * * * *
(b) Exemption from the requi rement of a tolerance is
established for residues of the microbial insecticide Bacillus thuringiensis Berliner, as specified in paragraph (a) of this section, 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. The Office of Pesticide Programs developed a standard commodity vocabulary database entitled Food and Feed Commodity Vocabulary

. The exemption from requirement of tolerance was updated to comport with

this new database.