### BIOPESTICIDES REGISTRATION ACTION DOCUMENT



## Bacillus pumilus strain BU F

- 33

Pesticide Chemical (PC) Code: 0 07493

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

DRAFT

– May 9 , 2013Bacillus pumilus strain BU F

- 33 Page 2 of 32 Biopesticides Registration Action Docum ent



### TABLE OF CONTENTS

I. EXECUTIVE SUMMARY

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

II. ACTIVE INGREDIENT OV ERVIEW

................................ ................................ ................................ ..... 7

III. REGULATORY BACKGROUN D

................................ ................................ ................................ ............ 7

A. Applications for Pesticide Product Registration

................................ ................................ ............ 7

B. Food Tolerance Exemption

................................ ................................ ................................ ............. 7

IV. RISK ASSESSMENT

................................ ................................ ................................ ................................ .. 8

A. Product Analysis Assessment (40 C.F.R. § 158.2120)

................................ ................................ ... 8

B. Human Health Assessment (40 C.F.R. § 158.2140)

................................ ................................ ....... 8

C. Environmental Assessment (40 C.F.R. § 158.2150)

................................ ................................ ..... 15

V. ENVIRONMENTAL JUSTIC E

................................ ................................ ................................ ............... 17

VI. RISK MANAGEMENT DECI SION

................................ ................................ ................................ ........ 18

VII. ACTIONS REQUIRED OF THE REGISTRANT

................................ ................................ ................. 19

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

................................ ................................ .... 20

IX. BIBLIOGRAPHY

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

A. Studies Submitted to Support the Bacillus pumilus strain BU F

- 33 Pesticide Product Registrations

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

B. Environmental Protection Agency Risk Assessment Memoranda

................................ ............. 23

C. Other References

................................ ................................ ................................ ............................ 23

APPENDIX A. MICROB IAL PESTICIDES DATA REQUIREMENTS

................................ ........................ 25

APPENDIX B. PESTICI DE PRODUCTS

................................ ................................ ................................ ........... 32Bacillus pumilus strain BU F

- 33 Page 3 of 32 Biopesticides Registration Action Docum ent



### BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

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

Science Reviews

Ibrahim S. Barsoum , Ph.D. Product Analysis, Human Health John L. Kough, Ph.D. Product Analysis, Human Health Shannon Borges Environmental Effects Annabel Waggoner Environmental Effects

Regulations

Kimberly Nesci Chief, Microbial Pesticides Branch Jeannine Kausch Regulatory Action LeaderBacillus pumilus strain BU F

- 33 Page 4 of 32 Biopesticides Registration Action Docum ent

I. EXECUTIVE SUMMARY

Background In January 201 2

, Becker Underwood , Inc.

, submitted application s for a manufacturing

- use p esticide product, Bacillus pumilus strain BU F

- 33 Technical ( EPA File Symbol : 71840

- RE )

, and an end

- use pesticide product

, Integral ® F

- 33 ( EPA File Symbol : 71840

- RG )

, to the U .S. Environmental Protection Agen cy (EPA) under section 3 of the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA)

. These pesticide products contain a new active ingredient, Bacillus pumilus strain BU F

- 33

.
Bacillus pumilus are Gram

- positive, endospore

- forming bacteria that are ubiquit ous in nature and have been identified and isolated from samples of a variety of media, including soil, water, aquatic i nvertebrates, and growing plants ( Garbeva et al . 2003; Halverson et al . 2000; Ivanova et al . 1999; Juhnke et al . 1987; Merina et al . 201 1; Oguntoyinbo 2007 ; Puente et al . 2004; U.S. EPA 2 01 2 c ; Yasinok et al . 2010 ). Since 2001, three isolates of Bacillus pumilus (i.e., Bacillus pumilus strain GB34, Bacillus pumilus strain QST 2808, and Bacillus pumilus strain GHA 180) have been used in regi stered pesticide products to suppress plant pathogens such as Pythium ultimum

, Fusarium oxysporum
, Rhizoctonia solani , and Alternaria species ( U.S. EPA 2 012c ). In conjunction with some of these registered pesticide products, the EPA established the followi ng exemptions from the requirement of a tolerance:

(1) Bacillus pumilus strain GB34 ( 40 C

. F

. R

. § 180.1224 )

– see the Federal Registers of April 2, 2003 ( 68 FR 15958 ) and December 22, 2004 ( 69 FR 76619 )

.
(2) Bacillus pumi lus strain QST 2808 ( 40 C

. F

. R

. § 180.1255 )

– see the Federal Register of November 3, 2004 ( 69 FR 63950 )

.
(3) Bacillus pumilus strain GHA 180 ( 40 C

. F

. R

. § 180.1313 )

– see the Federal Register of Ma rch 30, 2012 ( 77 FR 19109 )

.
Bacillus pumilus strain BU F

- 33 was isolated from inside the stem of a healthy cucumber plant a t an Auburn University facility outside Ta llassee, Alabama . Data demonstrate that Bacillus pumilus strain BU F

- 33 elicits systemic changes in plants, thereby enhancing their defense against a spectrum of pathogens, including viruses, fung i, and bacteria . T his increased protection results in more vigorous root and shoot systems in treated plants and leads to more unifor m stands and greater yields. In light of Bacillus pumilus strain BU F

- 33’s plant regulator capabilities, Becker Underwood, Inc.

, proposed to register a manufacturing

- use pesticide product and an end

- use pesticide product that is intended to be commercially applied to the seed of several plants (e.g., carrot and tomato) to elicit induced systemic resistance.

The EPA ’s s cientists reviewed produc t analysis, toxicology, and nontarget organism data and information ( 40 C

. F

. R

. §§ 158.2120, 158.2140, and 158.2150, respectively) submitted to support the registration of the two proposed Bacillus pumilus strain BU F

- 33 pesticide products. Overall, such da ta and information are adequate for risk assessment purposes , fulfill the current microbial pesticide data requirements

, and support registration of the proposed products under FIFRA section 3(c)(5)

.Bacillus pumilus strain BU F

- 33 Page 5 of 32 Biopesticides Registration Action Docum ent
Product Analysis The applicant fulfilled the product analysis data requirements for Bacillus pumilus strain BU F

- 33

, including product chemistry and composition, analysis of samples, and physical and chemical characteristics, by acceptable guideline studies.

Toxicology The applicant submitted a dequate mamm a lian toxicology data and information to support the Bacillus pumilus strain BU F

- 33 pesticide products. An a cute injection toxicity/pathogenicity (intravenous) study showed t hat, at a single high dose, Bacillus pumilus strain BU F

- 33 is not toxic , infectiv e, and/or pathogenic via this route of exposure

. Based on the applicant’s scientific rationale, which included citation to other toxicological studies done on Bacillus pumilus strain BU F

- 33 that resulted in no reporte d incidences of toxicity, infectivity, and/or pathogenicity, the EPA waived the requirement for
acute oral and acute pulmonary toxicity/pathogenicity testing. Moreover, the applicant reported that no hypersensitivity incidents occurred during rese arch, development, or testing of Bacillus pumil us strain BU F

- 33

. In l ight of the results of the acute toxicity/pathogenicity data and the absence of hypersensitivity incidents, the EPA is not requiring testing at higher tiers (i.e., Tiers II and III)

.
Tolerance Exemption The EPA concludes that there is a reasonable certainty that no harm will result to the U.S. population, including infants and children, from aggregate exposure to residues of Bacillus pumilus strain BU F

- 33

. The EPA expects no dietary risks are expected from use of Bacillus pumilus st rain BU F

- 33 as an active ingredient in pesticide products. Although exposure to Bacillus pumilus strain BU F

- 33 through consumed food and d rinking water may be possible, the EPA anticipates these types of exposure s to be negligible because of (1) the appl ication method o f the proposed end

- use pesticide product (i.e., applications will be made to seed located inside of commercial seed treatment facilities ) ; (2) Bacillus pumilus strain BU F

- 33’s colonization of and growth with developing plant roots that are often far removed from locations where food commodities develop (e.g., apple and corn) ; (3 ) typical food commodity processing procedures (e.g., peeling, washing, and/or additions of sugar, salt, and other preservatives) that tend to remove microbes or ren der them nonviable (U.S. EPA 1996 a ) ; and (4 ) pesticide product label instructions that commercial seed treatment facilities must adhere to so as not to contaminate water with pesticides . Should Bacillus pumilus strain BU F

- 33 be present on food or in drink ing water, supporting toxicological d ata indicate that human exposure to this bacterium would not result in unreasonable adverse effects.

Occupational Exposure Despite the low toxicological profile of Bacillus pumilus strain BU F

- 33

, the EPA is requiring baseline personal protective equipment (PPE) for handlers who, due to their occupation, may be exposed to the active ingredient for prolonged periods and/ or numerous times . Handlers working with Bacillus pumilus strain BU F

- 33 in commercial seed treatment settings are directed to wear a long

- sleeved shirt, long pants, socks, shoes, waterproof gloves, and a dust/mist filtering respirator meeting National Institute for Occupational Safety and Health (NIOSH) standards of at least N

- 95, R

- 95, or P

- 95. For futur e products, the EPA may require additional PPE (e.g., protective eyewear) , other than the standard described above, on a product

- specific basis.Bacillus pumilus strain BU F

- 33 Page 6 of 32 Biopesticides Registration Action Docum ent
Nontarget Organisms The EPA found that s cientific rationale and an acute avian oral toxicity/pathogenicit y study submitted by the applicant are sufficient to fulfill the nontarget organism data requirements and for risk assessment purposes. Further testing of nontarget organisms at higher tier levels (i.e., Tiers II, III, and IV) is not required. The EPA perf ormed an environment al risk assessment

, based on information provided by the
ap plicant

, and determined that unreasonable adverse effects to nontarg et organisms are not expected from the proposed pesticidal uses of Bacillus pumilus strain BU F

- 33 . Moreover, the EPA made a “No Effect” determination for direct and indirect effects to federally listed threatened and endangered species and their d esignated critical habitats resulting from these same proposed pesticidal uses.

Public Participation On Octob er 1, 2009, the EPA announced a policy to provide a more meaningful opportunity for the public to participate i n major registration decisions before they occur. According to this policy, the EPA provides a public comment period prior to making a registration decision for the following types of appli cations: new active ingredients; first food uses; first outdoor uses; first residential use s; or any other registration actions for which the EPA believes there may be significant public interest

.

Consistent with the policy of making registration actions more transparent, the proposed pe sticide products containing Bacillus pumilus strain BU F

- 33

, a new active ingredient, will be subject to a comment period. In addition to containing a new active ingredient, the re gistration of these pesticide products would result in the first food and outdoor use s for Bacillus pumilus strain BU F

- 33

. The docket identification number, which is associated with these registration actions and can be accessed through either http://www.regulations.gov or http://www.epa.gov/pesticides/regulating/registration

- status.html

, is EPA

- HQ

- OPP

- 201 2

- 0 265 . The following docum ents are available for comment in EPA

- HQ

- OPP

201 2

- 0 265 : (1) draft Bacillus pumilus strain BU F

- 33 Biopesticides Registration Action Document (BRAD); (2) environmental risk assessment for Bacillus pumilus strain BU F

- 33 ; (3) draft label for the manufacturin g

- use pesticide product, Bacillus pumilus strain BU F

- 33 Technical (EPA File Symbol : 71840

- RE ); and (4) draft label for the end

- use pesticide product, Integral ® F

- 33 (EPA File Sym bol : 71840

- RG ). While a final decision on registration is contingent upon rev iew and consideration of public comments, the EPA presently believes that, based upon the risk assessment and information submitted in support of Bacillus pumilus strain BU F

- 33 Technical and Integral ® F

- 33

, it is appropriate to issue these registrations. The basis for this preliminary decision can be found in the risk assessment for Bacillus pumilus strain BU F

- 33 , which is characterized throughout this BRAD.Bacillus pumilus strain BU F

- 33 Page 7 of 32 Biopesticides Registration Action Docum ent

II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Bacillus pumilus strain BU F

- 33

Culture Deposit: Agricultural Research Service Culture Collection (also known as the Northern Regional Research Laboratory (NRRL) Collection) in Peoria, Illinois under Accession Number B

- 50185

OPP Chemical Code: 0 07493

Type of Pesticide: Microbial Pesticide

– Plant Regulator

See Appendix B for specific information (e.g., use sites, application rate s , methods of application, formulation type s , and target pests) regarding the proposed p esticide products containing this active ingredient.

III. REGULATORY BACKGROUND

A. Applications for Pesticide Product Registration

In January 2012

, Becker Underwood , Inc. (address: 801 Dayton Avenue ; P.O. Box 667; Ames, Iowa 50010 )

, submitted applic ation s to register a manufacturing

- use pesticide product, Bacillus pumilus strain BU F

- 33 Technical (EPA File Symbol: 71840

- RE )

, and an end

- use pesticide product, Integral ® F

- 33 (EPA File Symbol: 71840

- RG )

, under FIFRA section 3. On June 27, 2012 ( 77 FR 38285 )

, the EPA announced receipt of these application s to register pesticide product s containing a new active ingredient and opened a 3 0

- day public comment period pursuant to the provisions of FIFRA section 3(c)(4). No

comme nts were received following th is publication.

B. Food Tolerance Exemption

Concurrent with its registration application s and under Federal Food, Drug, and Cosmetic Act (FFDCA) section 408(d), Becker Underwo od, Inc.

, submitted a petition to establish an exemption from the requirement of a tolerance for Bacillus pumilus strain BU F

- 33 ( PP 2F7978 )

. In the Federal Register of September 28, 2012 ( 7 7 FR 59578 ), the EPA announced that Becker Underwood, Inc.

, proposed to establish an exemption from the requirement of a tolerance for residues of the elicitor of induced systemic resistance

, Bacillus pumilus strain BU F

- 33 , in or on all food commodities and opened a 30

- day comment period. No comme nts were received following this publication.Bacillus pumilus strain BU F

- 33 Page 8 of 32 Biopesticides Registration Action Docum ent

IV. RISK ASSESSMENT

In the Federal Register s of October 26, 2007 ( 7 2 FR 60988 ) and August 30, 2012 ( 77 FR 52610 )

, the EPA issued Final Rule s on the data requirements to support registration of microbial pesticides and updated the definition fo r microbial pesticides

. The data and information evaluated for this BRAD were considered in light of these requirements.

The classifications that are found for each data submission are assigned by the EPA ’s science reviewers and are an indication of the u sefulness of the information contained in the documents for risk assessment. A rating of “ acceptable ” indicates the study is scientifically sound and is useful for risk assessment. A “ supplemental ” rating indicates the data provide some information that ca n be useful for risk assessment. The studies may have certain aspects determined not to be scientifically acceptable (“ supplemental : upgradeable ”). If a study is rated as “supplemental: upgradeable ,” the EPA always provides an indication of what is lacking or what can be provided to change the rating to “ acceptable .” If there is simply a “ supplemental ” rating, the reviewer will often state that t he study is not required by 40 C

. F

. R

. P art 158. Both “ acceptable” and “supplemental ” studies may be used in the r isk assessment
process as appropriate. An “unacceptable ” rating indicates that new data must be submitted.

For the acu te toxicity data requirements, Toxicity C ategories are assigned based on the hazard(s) identified from studies and/or other infor mation submitted to the EPA in support of a pesticide registration. The active ingredient or particular product is classified into Toxicity Category I, II, III, or IV, where Toxicity Category I indicates the highest toxicity and Toxicity Category IV indicates the lowest toxicity (see section II of U.S. EPA ( 2012d ) )

.
A. Product Analysis Assessment ( 40 C

. F

. R

. § 158 .2120 )

The applicant fulfilled a ll product analysis data requirements for Bacillus pumilus strain BU F

- 33 . Refer to Tables 1, 2, 3, and 4 in Appendix A for a summary of the data requirements, including both generic and prod uct

- specific information.

B. Human Health Assessment ( 40 C

. F

. R

. § 158.2140 )

1. Toxicity

The applica nt fulfilled a ll toxicology data requirements for Bacillus pumilus strain BU F

- 33

. Acceptable Tier I mammalian toxicology data and information support registration of the Bacillus pumilus strain BU F

- 33 pesticide products

. The EPA is not requiring Tier II and Tier III studies for Bacillus pumilus strain BU F

- 33 based on the lack of acute toxicity/pathogenicity in the Tier I studies attributed to the bacterium

.
For a summary of the generic toxicology data requirements described in sections IV(B)(1)(a) and I V(B)(1)(b), as well as additional product

- specific data submitted to support the individual registrations, refer to Tables 5 and 6 in Appendix A

.Bacillus pumilus strain BU F

- 33 Page 9 of 32 Biopesticides Registration Action Docum ent

a. Acute Toxicity/Pathogenicity

– Tier I

Acute Oral Toxicity/Pathogenici ty ( Harmonized Guideline 885.3050; Master Record Identification Number (MRID No.) 487164

- 10 ) : Upon consideration of results of other definitive toxicologic al data submitted by the applicant

, the EPA waived acute oral toxicity/pathogenicity testing for Baci llus pumilus strain BU F

- 33. An acute oral toxicity study conducted on female rats (MRID No. 487164

- 03) demonstrated that Bacillus pumilus strain BU F

- 33 was not toxic (LD 50 greater than 5,000 mg/kg; Toxicity Category IV), while an acute injection toxicity /pathogenicity study conducted on rats (MRID No. 487164

- 09) showed that Bacillus pumilus strain BU F

- 33 was not toxic, infective, and/or pathogenic when the skin was bypassed as a barrier. The EPA believes these data, when taken together, indicate that thi s bacterium would not be toxic, infective, and/or pathogenic through the oral route of exposure and that further testing is not necessary

.

Acute Pulmonary Toxicity/Pathogenicity ( Harmonized Gui deline 885.3150; MRID No. 487164

- 10 ) : Upon consideration of re sults of other definitive toxicological data submitted by the applicant

, the EPA waived acute pulmonary toxicity/pathogenicity testing for Bacillus pumilus strain BU F

- 33. An acute inhalation toxicity study conducted on rats (MRID No. 487164

- 05) demonstrat ed that Bacillus pumilus strain BU F

- 33 was not toxic (LC 50 greater than 2.22 mg/L; Toxicity Category IV), while an acute injection toxicity/pathogenicity study conducted on rats (MRID No. 487164

- 09) showed that Bacillus pumilus strain BU F

- 33 was not toxi c, infective, and/or pathogenic when the skin was bypassed as a barrier. The EPA believes these data, when taken together, indicate that this bacterium would not be toxic, infective, and/or pathogenic through the inhalation route of exposure and that furth er testing is not
necessary

.
Acute Injection Toxicity/Pathogenicity (Intravenous)

– Rat (Harmonized Guideline 885.3200; MRID No. 487164

- 02) : In an acute intravenous injection toxicity and pathogenicity study, fifteen male and fifteen female young adult Sp rague

- Dawley rats were injected intravenously with Bacillus pumilus strain BU F

33 (6.38 x 10 11 viable spores per gram) in sterile phosphate buffered saline at a dose of 6.1 x 10 10 cfu/animal (0.1 milliliter dosing volume). The animals were treated on day 0 and observed for up to 21 days, with interim sacrifices (3 animals per sex) takin g place on days 0, 3, 7, and 14 and terminal sacrifice (3 animals per sex) on day 21. Five males and five females were maintained for 21 days as untreated controls, and five males and five females were treated on day 0 with intravenous inactive (autoclaved) Bacillus pumilus strain BU F

- 33 in sterile phosphate buffered saline (0.1 milliliter) and sacrificed on day 21.

There were no treatment

- related deaths or clinical signs. Four of the six active

- treated animals killed on day 3 had small weight losses of 1

– 4 grams during days 0

– 3; otherwise, all of the animals gained weight throughout the observation period. I nactive

- treated and active

- treated males , however, had decreased

c umulative weight gain relative to untreated controls ( 20

– 21% less) , which corresponded to reduced mean absolute body weights (

- 9% and

- 11% for inactive

- and active

- treated animals, respectively). Black and/or enlarged spleens were noted in one male and thr ee females in the inactive

- treated group. Dark or black and/or enlarged spleens were noted in active

- treated animals as follows: one female killed on day 3, one female killed on day 7, three males and two females killed on day 14, and one male and three fe males killed on day 21. Enlarged lymph nodes were an additional finding in one of the males killed on day 14. No abnormal gross necropsy findings were seen in controls. Correlated organ weight changes included i ncreased absolute mesenteric lymph node weigh t in inactive

- and active

- treated animals ofBacillus pumilus strain BU F

- 33 Page 10 of 32 Biopesticides Registration Action Docum ent

both sexes (males: +19% and +78%; females: +15% and +21%, respectively , relative to untreated controls ) and increased absolute spleen weight in inactive

- and active

- treated animals of both sexes (males: +84% and +105%; females: +81% and +84%, respectively). The test organism was cleared from the blood, kidney, brain, and lungs by day 21; however, the test organism remained present in the liver, spleen, mesenteric lymph nodes, and cecum contents on day 21.

Based on the results of this study

, Bacillus pumilus strain BU F

- 33 dosed intravenously at 6.1 x 10 10
cfu /animal does not appear to be infective or pathogenic in rats ; however, toxicity cannot definitely be ruled out. This study was rated acceptable.

Regarding the toxicity uncertainty mentioned above, i t is known that intravenous injection with a high dose of bacteria can cause toxicity in animals, and, in this study, the test animals were dosed three orders of magnitude higher than the test guideline requires (6.1 x 10 10 vs

. 1 x 10 7 cfu/animal , see U.S. EPA ( 1996b ) ) . Thus, these adverse effects were likely caused by high bacterial numbers rather than toxicity attributed to Bacillus pumilus strain BU F

- 33 . To rule out toxicity, a lower dose (5.2 x 10 7 cfu/animal ) in the range recommended by U.S. EPA (1996b) w as tested in MRID No. 487164

- 09. This particular test de monstrated that Bacillus pumilus strain BU F

- 33 was not toxic, infective, and/or pathogenic in rats (see directly below for more details about this stud y ).

Acute Injection Toxicity/Pathogenicity (Intravenous)

– Rat (Harmonized Guideline 885.3200; MRID No. 487164

- 09) : In an acute intravenous injection toxicity and pathogenicity study, eighteen male and eighteen female young adult Sprague

- Dawley rats were injected with Bacillus pumilus strain BU F

- 33 (6.38 x 10 11 viable spores per gram) in sterile phosphate buffered saline at a dose of 5.2 x 10 7 cfu/animal. The animals were treated on day 0 and observed for up to 42 days, with interim sacrifices (3 animals per sex) taking place on days 0, 14, 21, 28, and 35 and terminal sacrifice (3 animals per sex) on day 42. Two males and two females were maintained for 42 days as untreated controls.

There were no treatment

- related deaths, clinical signs, gross necropsy findings, or changes in body weight or body weight gain. No test organism was recovered from brains of the treated animals. The test organism was found in the blood on day 0 (only) and in the lung, spleen, liver, kidney, mesenteric lymph node, and cecum co ntent samples taken on day 14. The test organism was cleared (i.e., two consecutive sacrifices with all values less than 100 cfu/g) from the liver by day 28; from the spleen, kidney, and mesenteric lymph nodes by day 35; and from the cecum content by day 4 2. For the lungs, all values were less than 100 cfu/g on day 28 (only), and no further tests were done on lung tissue thereafter.

Based on the results of this study, Bacillus pumilus strain BU F

- 33 dosed intravenously at 5.2 x 10 7
cfu/animal does not app ear to be toxic, infective, and/or pathogenic in rats. This study was rated acceptable.

Hypersensitivity Incidents ( Harmonized Guideline 885.3400; MRID No. 487164

- 10 ) : The applicant reported that n o hypersensitivity incidents, including immediate

- type or delayed

- type reactions of humans and domestic animals

, occurred during research, development, or testing of Bacillus pumilus strain BU F

- 33 . Any future hypersensitivity incidents must be reported to the EPA (refer to test note #3 of 40 C

. F

. R

. § 158.214 0(d))

.
Cell Culture ( Harmonized Guideline 885.3500 ; MRID No. 487164

- 10 ) : This study is not required because Bacillus pumilus strain BU F

- 33 is not a virus (refer to test note #4 of 40 C

. F

. R

. § 158.2140(d)).Bacillus pumilus strain BU F

- 33 Page 11 of 32 Biopesticides Registration Action Docum ent

b. Acute Toxicology and Subchronic Toxicity/ Pathogenicity

– Tier II; Reproductive Fertility Effects, Carcinogenicity, Immunotoxicity, and Infectivity/Pathogenicity Analysis

– Tier III

The EPA is not requiring Tier II and Tier III studies for Bacillus pumilus strain BU F

- 33 based on the lack of acute toxicity/pathogenicity in the Tier I studies attributed to the bacterium

. Any uncertainties about the toxicity of Bacillus pumilus strain BU F

- 33 raised in one acute injection toxicity/pathogenicity study (MRID No. 487164

- 02) were likely as a result of dosing, not actual toxicity, as demonstrated by a subsequent acute injection toxicity/pathogenicity study (MRID No. 487164

- 09)

c. Endocrine Disruptors

As required by the Administrator under FFDC A section 408(p), the EPA has developed the Endocrine Disru ptor 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 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, the EPA issued test orders/data call

- ins for the first group of 67 chemicals, which contains 58 pesticide active ingredients and 9 inert ingredients. Bacillus pumilus

s train BU F

- 33 is not among the group of 58 pesticide active ingredients on the initial list to be screened under the EDSP.

The EPA believes tha t Bacillus pumilus strain BU F

- 33 , the active ingredient described in this document, 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, pu rsuant to Section 408(p)(4), the EPA will determine in the future whether it can exempt Bacillus pumilus strain BU F

- 33 from the requirements of the Section 408(p) EDSP. In the event the EPA 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, the t est guidelines and the Tier 1 screening battery, please visit our websit e at http://www.epa.gov/endo/

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

Section 408(c)(2)(A)(i) of FFDCA allows the E PA to establish an exemption from the requirement for a tolerance (the legal limit for a pesticide chemical residue in or on a food) only if the EPA determines that the exemption is “safe.” Section 408(c)(2)(A)(ii) of FFDCA defines “safe” to mean that “the re is a reasonable certainty that no harm will result from aggregate exposure to the pesticide chemical residue, including all anticipated dietary exposures and all other exposures for which there is reliable information.” This includes exposure through dr inking water and in residential settings but does notBacillus pumilus strain BU F

- 33 Page 12 of 32 Biopesticides Registration Action Docum ent
include occupational exposure. Pursuant to FFDCA section 408(b)(2)(B) , in establishing or maintaining in effect an exemption from the requirement of a tolerance, the EPA must take into account the facto rs set forth in FFDCA section 408(b)(2)(C), which require the EPA to give special consideration to exposure of infants and children to the pesticide chemical residue in establishing a tolerance exemption

, and to “ensure that there is a reasonable certainty that no harm will result to infants and children from aggregate exposure to the pesticide chemical residue....” Additionally, FFDCA section 408(b)(2)(D) requires that the EPA consider “available information concerning the cumulative effects of [a particul ar pesticide's] . . . residues and other substances that have a common mechanism of toxicity.”

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

Consistent with FFDCA section 408(b)(2)(D), the EPA reviewed the available scient ific data and other relevant information on Bacillus pumilus strain BU F

- 33 , and considered its validity, completeness, and reliability, as well as the relationship of this information to human risk. The EPA also considered available information concerning the variability of the sensitivities of major identifiable subgroups of consumers, including infants and children.

Based on the acute toxicity/pathogenicity data and information discussed previously and presented in Table s 5 and 6 in Appendix A , t he data required for a FFDCA risk assessment for Bacillus pumilus strain BU F

- 33 have been fulfilled.

a. Aggregate Exposure

In examining aggregate exposure, FFDCA section 408 directs the EPA to consider available information

con cerning exposures from the pesticide residue in food and all other non

- occupational exposures, including drinking water from ground water or surface water and exposure through pesticide use in gardens, lawns, or buildings (residential and other indoor uses ).

Food Exposure and Risk Characterization : Exposure to residues of Bacillus pumilus strain BU F

- 33, a naturally occurring bacterium ( Garbeva et al . 2003; Halverson et al . 2000; Ivanova et al . 1999; Juhnke et al . 1987; Merina et al . 2011; Oguntoyinbo 2007 ; Puente et al . 2004; U.S. EPA 2012c; Yasinok et al

. 2010 ) , in consumed food is anticipated to be possible but negligible. Bacillus pumilus strain BU F

- 33 is to be applied to various plant seeds within commercial seed treatment facilities. Subsequently, tr eated seeds would be bagged, sold to growers, and likely deposited into soil in agricultural settings. Since Bacillus pumilus strain BU F

- 33 colonizes and grows with developing plant roots, it is possible that food commodities that develop underground (e.g ., root and tuber vegetables) or in close proximity to the soil/ground (e.g., cucumber and pumpkin) could be directly exposed to Bacillus pumilus strain BU F

- 33. Further, treated seed could be diverted for food or feed purposes or processed into oil; there fore, exposure to Bacillus pumilus strain BU F

- 33 in this situation is also plausible. Although residues from the use of pesticides containing Bacillus pumilus strain BU F

- 33 may be present on certain food commodities, processing procedures (e.g., peeling and washing) may reduce such residues ( U.S. EPA 1996 a ). Regardless of the scenario, should Bacillus pumilus strain BU F

- 33 be present on food, supporting toxicological data indicate that no toxicity, pathogenicity, and/or infectivity is likely to occur wit h any food exposure resulting from the use of Bacillus pumilus strain BU F

- 33 as a pesticide whenBacillus pumilus strain BU F

- 33 Page 13 of 32 Biopesticides Registration Action Docum ent
applied in accordance with label directions and good agricultural practices (see section IV(B)(1)(a) and Table s 5 and 6 in Appen dix A ).

Drinking Water Exposure and Risk Characterization : Like consumed food, exposure to residues of Bacillus pumilus strain BU F

- 33, a naturally occurring bacterium ( Garbeva et al . 2003; Halverson et al

. 2000; Ivanova et al . 1999; Juhnke et al . 1987; M erina et al . 2011; Oguntoyinbo 2007 ; Puente et al

. 2004; U.S. EPA 2012c; Yasinok et al . 2010 ) , in consumed drinking water is anticipated to be possible but negligible. The proposed use pattern for Bacillus pumilus strain BU F

- 33 involves commercial seed
tr eatment that takes place indoors. Commercial seed treaters are required to avoid contamination of water when disposing of equipment washwaters or rinsate and, for applications involving container sizes greater than or equal to 5 gallons or 50 pounds, to ob tain a National Pollutant Discharge Elimination System permit and to notify the permitting authority in writing prior to discharge into lakes, streams, ponds, estuaries, oceans, and other waters (see sections II, III, and VII of U.S. EPA ( 2012e ) ). Thus, fo r larger container sizes, it is possible that commercial seed treatment facilities could discharge effluent containing Bacillus pumilus strain BU F

- 33 into surface waters. Additionally, Bacillus pumilus strain BU F

- 33 on treated seed that is planted by gro wers will colonize and grow with the roots of developing plants; therefore, its association with plant roots in the soil may result in exposure to ground and surface waters. If Bacillus pumilus strain BU F

- 33 were to be transferred to ground or surface wat ers as a result of effluent discharged from commercial seed treatment facilities or treated seed planted by growers, it is possible that this bacterium could survive the conditions water is subjected to in wastewater treatment systems and drinking faciliti es (e.g., pH adjustments and chlorination). According to the World Health Organization, Bacillus species, such as pumilus , are often detected in drinking water even after going through acceptable water treatment processes, largely because the spores are re sistant to these disinfection processes ( World Health Organization 2011 ). In the remote likelihood that Bacillus pumilus strain BU F

- 33 is present in drinking water, supporting toxicological data indicate that no toxicity, pathogenicity, and/or infectivity is likely to occur with any drinking water exposure resulting from the use of Bacillus pumilus strain BU F

- 33 as a pesticide when applied in accordance with label directions and good agricultural practices (see section IV(B)(1)(a) and Table s 5 and 6 in Appendix A ).

Non

- occupational, Residential Risk Characterization : Given Bacillus pumilus ’ natural presence in the environment ( Garbeva et al . 2003; Halverson et al . 2000; Ivanova et al . 1999; Juhnke et al . 1987; Merina et al

. 2011; Oguntoyinbo 2007 ; Puente et al . 2004; U.S. EPA 2012c; Yasinok et al . 2010 )

, dermal and inhalation non

- occupational exposures to Bacillus pumilus strain BU F

- 33 almost certainly are already occurring. Additional exposure to Bacillus pumilus strain BU F

- 33 due to pesticidal applications is not expected because the proposed end

- use pesticide product is labeled only for use inside of commercial seed treatment facilities. Subsequently, treated seeds would be bagged, sold to growers, and likely deposited in to soil in agricultural settings; diverted for food or feed purposes; or processed into oil. Even if additional non

- occupational exposures were to occur (e.g., through eventual expansion of use sites or use patterns), no adverse effects attributed to Bacil lus pumilus strain BU F

- 33 are anticipated, particularly in light of testing that indicated that this bacterium is not toxic (acute inhalation toxicity and acute dermal toxicity tests), is only slightly irritating (primary dermal irritation test), and is

n ot pathogenic or infective (acute injection toxicity/pathogenicity test) at high doses ( see section IV(B)(1)(a) and Tables 5 and 6 in Appendix A ).Bacillus pumilus strain BU F

- 33 Page 14 of 32 Biopesticides Registration Action Docum ent

b. Cumulative Effects from Substances with a Common Mechanism of Toxici ty

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

The EPA has not found Bacillus pumilus strain BU F

- 33 to share a common mechanism of toxicity with any other substances, and Bacillus pumilus strain BU F

- 33 does not appear to produce a toxic metabolite
pro duced by other substances. For the purposes of this tolerance action, therefore, the EPA has assumed that Bacillus pumilus strain BU F

- 33 does not have a common mechanism of toxicity with other substances. Following from this, therefore, the EPA concludes that there are no cumulative effects associated with Bacillus pumilus strain BU F

- 33 that need to be considered. For information regarding the EPA’s efforts to determine chemicals that have a common mechanism of toxicity and to evaluate the cumulative effe cts of such chemicals, see the EPA’s website at http://www.epa.gov/pesticides/cumulative

.

c. Determination of Safety for the United States Population, Infants and Children

FFDCA section 408(b)(2 )(C) provides that, in considering the establishment of a tolerance or tolerance exemption for a pesticide chemical residue, the EPA shall assess the available information about consumption patterns among infants and children, special susceptibility of inf ants and children to pesticide chemical residues, and the cumulative effects on infants and children of the residues and other substances with a common mechanism of toxicity. In addition, FFDCA section 408(b)(2)(C) provides that the EPA shall apply an addi tional tenfold (10X) margin of safety for infants and children in the case of threshold effects to account for prenatal and postnatal toxicity and the completeness of the database on toxicity and exposure, unless the EPA determines that a different margin of safety will be safe for infants and children. This additional margin of safety is commonly referred to as the Food Quality Protection Act Safety Factor. In applying this provision, the EPA either retains the default value of 10X, or uses a different add itional or no safety factor when reliable data are available to support a different additional or no safety factor.

Based on the acute toxicity an d pathogenicity data/information discussed in section IV(B)(1)(a) and Tables 5 and 6 in Appendix A

, the EPA concludes that there are no threshold effects of concern to infants, children, or adults when Bacillus pumilus strain BU F

- 33 is used as labeled in accordance with good agricultural practices. As a result, the EPA concludes t hat no additional margin of exposure (safety) is necessary.

Moreover , based on the same data and analysis as presented directly above, the EPA is able to conclude that there is a reasonable certainty that no harm will result to the U.S. population, includ ing infants and children, from aggregate exposure to the residues of Bacillus pumilus strain BU F

- 33 when it is used as labeled and in accordance with good agricultural practices as an elicitor of induced systemic resistance in plants. Such exposure includ es all anticipated dietary exposures and all other exposures for which there is reliable information. The EPA has arrived at this conclusion because, considered collectively, the data and information available on Bacillus pumilus strain BU F

- 33 do not demo nstrate toxic, pathogenic, and/or infective potential to mammals, including infants and children.Bacillus pumilus strain BU F

- 33 Page 15 of 32 Biopesticides Registration Action Docum ent

3 . Occupational Exposure and Risk Characterization

Handler exposure to Bacillus pumilus strain BU F

- 33 is not expected to pose any undue risk. Regardless, the EPA is requiring appropriate personal protective equipment and precautionary statements on pesticide product labels to mitigate any potential risks to pesticide handlers due to prolonged and/ or numerous exposures. Handlers working with Bacillus pumilu s strain BU F

- 33 in commercial seed
treatment settings are directed to wear a long

- sleeved shirt, long pants, socks, shoes, waterproof gloves, and a dust/mist filtering respirator meeting National Institute for Occupational Safety and Health (NIOSH) standa rds of at least N

- 95, R

- 95, or P

- 95. For future products, the EPA may require additional PPE (e.g., protective eyewear) , other than the standard described above, on a product

- specific basis.

4 . Human Health Risk Characterization

The EPA considered huma n exposure to Bacillus pumilus strain BU F

- 33 in light of the standard for registration in FIFRA and the relevant safety factors in FFDCA. A determination has been made that no unreasonable adverse effects to the U.S. population in general, and to infants and children in particular, will result from incidental exposures when Bacillus pumilus strain BU F

- 33 pesticide products are applied to seed in accordance with the EPA’s approved labeling.

C. Environmental Assessment ( 40 C

. F

. R

. § 158.2150 )

The EPA found that scientific rationale and an acute avian oral toxicity/pathogenicity study submitted by the applica nt are sufficient to fulfill the nontarget organism data requirements and for risk assessment purposes. Further testing of nontarget organisms at higher tier levels (i.e., Tiers II, III, and IV) is not required. The EPA performed an environment al risk asse ssment, based on information provided by the
ap plicant, and determined that unreasonable adverse effects to nontarget organisms are not expected from the proposed pesticidal uses of Bacillus pumilus strain BU F

- 33 . Moreover, the EPA made a “No Effect” dete rmination for direct and indirect effects to federally listed threatened and endangered species and their designated critical habitats resulting from these same proposed pesticidal uses.

For a summary of the generic data requirements described in secti ons IV(C)(1), refer to Table 7 in Appendix A

.

1. Ecological Exposure and Risk Characterization

Since the proposed registration of the end

- use pesticide product is for commercial seed treatment only, exposure to nontarget organisms is expected to be minimal in most cases. Birds and mammals may be exposed through consumption of treated seed. Soil

- dwelling animals, including invertebrates, may also be exposed as a result of the presence of treated seed in their environment, though the opportunity for exposure in this case is expected to be limited. While Bacillus pumilus strain BU F

- 33 is likely to increase in population as the seed germinates and roots grow, its close association with plant roots should limit the exposure pr ofile within the soil to areas around plants growing from treated seed. Exposure to Bacillus pumilus strain BU F

- 33 in soil is not expected to be significant.Bacillus pumilus strain BU F

- 33 Page 16 of 32 Biopesticides Registration Action Docum ent

a. Birds (Harmonized Guidelines 885.4050 and 885.4100) and Wild Mammals (Harmonized Guideline 885.4150) (MRID Nos. 487164

- 08 and 487164

- 10)

As noted above, birds and mammals may be exposed to Bacillus pumilus strain BU F

- 33 through consumption of treated seeds. Data are available, however, to show that exposure is not likely to caus e adverse effects in these animals. An acute avian toxicity study with Northern bobwhite ( Colinus virginianus , MRID No. 487164

- 08) show ed no toxicity or pathogenicity due to Bacillus pumilus strain BU F

- 33 ( concentration of 6.38 x 10 11 cfu /g)

, whe n orally administered at 5

, 000 mg/kg b ody weight ( 3.19 x 10 12 cfu/kg b ody w eight ) daily for five consecutive days ( 1.6 x 10 13 cfu/kg b ody w eight total)

. Scientific rationale submitted to satisfy the avian inhalation toxicity/pathogenicity study also highlighted the influence of the use pattern in limiting inhalation exposure. Several studies are also available for determining the potential hazard to wild mammals. Bacillus pumilus strain BU F

- 33 was not infective or pathogenic to laboratory rats when injected with 6. 1 x 10 10 cfu/animal (MRID No. 487164

- 02) and was not infective, pathogenic, and/ or toxic to rats when injected at 5.2 x 10 7 cfu/animal (MRID No. 487164

- 09) . Additionally, Bacillus pumilus BU F

- 33 was not toxic in an acute oral toxicity study with rats
expo sed to 5,000 mg/kg by gavage (MRID No. 487164

- 03) . Rationale submitted to satisfy the wild mammal testing data requirement also cited a study in which Bacillus pumilus was fed to swine in the diet for ten days as a potential probiotic. Th is study indicat ed that the Bacillus pumilus did not colonize (or infect) the digestive tract s of the test animals (Kornegay and Risley 1996)

. Therefore, based on the studies and other information presented, risk to birds and wild mammals is expected to be minimal as a

resu lt of the proposed registrations of the Bacillus pumilus strain BU F

- 33 pesticide products

.

b. Nontarget Plants (Harmonized Guideline 885.4300) (MRID No. 487164

- 10)

Scientific rationale submitted to satisfy the data requirement for nontarget plants focused primarily on the lack of potential exposure to nontarget plants, particularly since the proposed labeled use pattern is for commercial seed treatment only. An additional report of Bacillus pumilus and its association w ith rice seed was also include d. In this study, Bacillus pumilus was associated with signs of sheath necrosis in seedlings but was considered to be of low pathogenic potential based on the criteria defined in the study (Cottyn et al . 2001) . The proposed use pattern is likely to minimiz e the potential for Bacillus pumilus BU F

- 33 to contact naturally growing nontarget plants or crop plants not treated with Bacillus pumilus BU F

- 33 as seed; however, a previous EPA review cited several cases in which Bacillus pumilus was shown to be pathog enic to crop plants (U

. S

. EPA 2013 c ), which may indicate concern for crop plants treated (as seed) with Bacillus pumilus strain BU F

- 33 . Based on this review, Becker Underwo od, Inc.

, revised the proposed end

- use pesticide product label to prohibit applicat ion of Bacillus pumilus strain BU F

- 33 to seeds of the crops indicated in the review (rice, potato, sweet potato, bean, cabbage, garlic, and pear). Therefore, due to the lack of exposure to nontarget plants and crops of concern, adverse effects to nontarge t plants are not expected from the proposed use of Bacillus pumilus strain BU F

- 33.

c. Nontarget Insects (Harmonized Guideline 885.4340) and Honey Bees (Harmonized Guideline 885.4380) (MRID No. 487164

- 10)

Exposure is expected to be minimal to non target insects and honey bees, due to the proposed use pattern as a commercial seed treatment. Additionally, Bacillus pumilus is naturally found in many environments, and insects and honey bees are likely to be exposed to Bacillus pumilus naturally via con tact with soil and water

. Therefore, based on minimal expected exposure, adverse effects to nontarget insects and honey bees are not anticipated as a result of the proposed registrations of the Bacillus pumilus strain BUBacillus pumilus strain BU F

- 33 Page 17 of 32 Biopesticides Registration Action Docum ent

F

- 33 pesticide products.

d. F reshwater Fish (Harmonized Guideline 885.4200), Freshwater Invertebrates (Harmonized Guideline 885.4240), and Estuarine/Marine Fish and Invertebrates (Harmonized Guideline 885.4280) (MRID No. 487164

- 10)

As with other taxa discussed above, exposure of Bacillus pumilus strain BU F

- 33 in aquatic environments is expected to be minimal. Use as a commercial seed treatment will limit the presence of Bacillus pumilus strain BU F

- 33 to areas around the treated seed and growing plant. This use pattern present s little opportunity for runoff into nearby aquatic areas. Therefore, based on the expected lack of exposure, adverse effects to fish and aquatic invertebrates in freshwater, marine, and estuarine environments are not expected.

e. Conclusions for Non target Organisms

Based on the expected lack of exposure and data demonstrating no effects, as discussed above, unreasonable adverse effects to n ontarget organisms are not anticipated as a result of the proposed registration s of the Bacillus pumilus s train BU F

- 33 Technical and INTEGRAL ® F

- 33 pesticide products , containing Bacillus pumilus strain BU F

- 33, as labeled.

2. Environmental Fate Data

As t he rationale and data provided are sufficient to fulfill the Tier I nontarget organism data requirements a nd allow for nontarget organism risk asse ssment for Bacillus pumilus strain BU F

- 33 , the EPA is not requiring testing at higher tier levels (i.e., Tiers II, III, and IV)

.

3

. Threatened and Endangered Species Assessment

Since the EPA has determined tha t no effects are anticipated for nontarget species exposed to Bacillus pumilus strain BU F

- 33 a s a result of the proposed labeled use s, effects to federally listed threatened and endangered species and their designated critical habitats are also not expect ed. Therefore, a “No Effect” determination is made for direct and indirect effects to federally listed threatened and endangered species and their designated critical habitats for the proposed pesticidal use s of Bacillus pumilus strain BU F

- 33

, as labeled.

V. ENVIRONMENTAL JUSTICE

The EPA seeks to achieve environmental justice

— the fair treatment and meaningful involvement of all people regardless of race, color, national origin, or income

— with respect to the development, implementation, and enforcement of environmental laws, regulations, and policies. Fair treatment means that no group of people, including racial, ethnic, or socioeconomic groups, should bear a disproportionate share of the negative environmental consequences resulting from industrial, mu nicipal, and commercial operations or the execution of federal, state, local, and tribal environmental programs and policies. Meaningful involvement means that (1) potentially affected community residents have an appropriate opportunity to participate in d ecisions about a proposed activity that will affect their environment and/or health; (2) the public’s contribution can influence the regulatory agency’s decision;Bacillus pumilus strain BU F

- 33 Page 18 of 32 Biopesticides Registration Action Docum ent
(3) the concerns of all participants involved will be considered in the decision

- making proce ss; and (4) the decision

- makers seek out and facilitate the involvement of those potentially affected. The EPA has this goal for all communities and persons across the United States.

To help address potential environmental justice issues, the EPA seeks i nformation on any groups or segments of the population who, as a result their location, cultural practices, or other factors, may have atypical, unusually high exposure to Bacillus pumilus strain BU F

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

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

.

V I

. RISK MANAGEMENT DECISION

Section 3(c)(5) of FIFRA permits for the registration of a pesticide provided that all the following determinati ons are made:

(1) Its composition is such as to warrant the proposed claims for it; (2) Its labeling and other material required to be submitted comply with the requirements of FIFRA; (3) It will perform its intended function without unreasonabl e adverse effects on the environment; AND (4) When used in accordance with widespread and commonly recognized practice, it will not generally cause unreasonable adverse effects on the environment.

To satisfy criterion 1, the EPA under stands that the Bacillus pumilus microbe is well

- known to elicit induced systemic response in plants, enhancing plants’ ability to defend against pathogens

. The EPA has no knowledge that would contradict the claims made on the Bacillus pumilus strain BU F

- 33 Technical and Integral ® F

- 33 label s, and such pesticide products are not expected to cause unreasonable adverse effects on the environment when used according to the ir respective label instructions. Criterion 2 is satisfied by the current product label s , as well as the data and information presented in this document. It is believed that the Bacillus pumilus strain BU F

- 33 pesticide products will not cause any unreasonable adverse effects on the environment, and Integral ® F

- 33 ( end

- use pesticide product)

, in particular, is likel y to elicit induced systemic resistance in plants as claimed, satisfying criterion 3. Criterion 4 is satisfied in that the Bacillus pumilus strain BU F

- 33 pesticide products are not expected to cause unreasonable adverse effects whe n used according to label instructions. Therefore, EPA believes Bacillus pumilus strain BU F

- 33 Technical and Integral ® F

- 33

, containing Bacillus pumilus strain BU F

- 33 as a new active ingredient, are eligible for registration under FIFRA section 3(c)(5) f or the labeled uses.Bacillus pumilus strain BU F

- 33 Page 19 of 32 Biopesticides Registration Action Docum ent

V I I

. ACTIONS REQUIRED OF THE REGISTRANT

A. Final Printed Labeling

Before releasing pesticide products containing Bacillus pumilus strain BU F

- 33 for shipment, the registrant is required to provide appropriate final printed lab eling to the EPA

.

B. Terms of Registration

No additional data are being required as terms of the Bacillus pumilus strain BU F

- 33 Technical registration.

The following item will be a term of the Integral ® F

- 33 registration :

Corrosion Characteristics ( Harmonized Guideline 830.6320) : Within one year of registration, Becker Underwood , Inc.

, must provide the results of a one

- year corrosion characteristics study.

C. Reporting of Adverse Effects and Hypersensitivity Incidents

Notwithstanding the informati on stated in the previous sections, it should be clearly understood that certain specific data are required to be reported to the EPA as a requirement for maintaining the federal registration for a pesticide product. A brief summary of these types of data are described below.

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

. F

. R

. § 158.2140(d).Bacillus pumilus strain BU F

- 33 Page 20 of 32 Biopesticides Registration Action Docum ent

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

BRAD Biopesticides Registration Action Document C

. F

. R

. Code of Federal Regulations cfu colony

- forming units cfu/g colony

- forming units per gram cfu/kg colony

- forming units per kilogram cPs centipoise EDSP Endocrine Disruptor Screening Program EP end

- use pesticide product EPA United States Envi ronmental Protection Agency FFDCA Federal Food, Drug, and Cosme tic Act FIFRA Federal Insecticide, Fungicide, and Rodenticide Act fl oz fluid ounce FR Federal Register gal gallon lb pound lb/ft 3 pounds per cubic foot LC 50 median lethal concentration. A statistically derived concentration of a substance that can be

ex pected to cause death in 50% of test animals. It is usually expressed as the weight of substance per weight or volume of water, air, or feed (e.g., mg/L, mg/kg, or ppm). LD 50 median lethal dose. A statistically derived single dose that can be expected to c ause death in 50% of the test animals when administered by the route indicated (oral, dermal, or inhalation). It is expressed as a weight of substance per unit weight of animal (e.g., mg/kg). mg/kg milligram s per kilogram mg/L milligrams per liter MP m anufacturing

- use pesticide product MRID No. Master Record Identification Number NIOSH National Institute for Occupational Safety and Health NRRL Northern Regional Research Laboratory OCSPP Office of Chemical Safety and Pollution Prevention OPP Office of P esticide Programs PC Code Pesticide Chemical Code PP Pesticide Petition PPE personal protective equipment
ppm parts per million TGAI technical grade of the active ingredient U.S. United StatesBacillus pumilus strain BU F

- 33 Page 21 of 32 Biopesticides Registration Action Docum ent

I X. BIBLIOGRAPHY

A. Studies Submitted to Supp ort the Bacillus pumilus strain BU F

- 33 Pesticide Product Registrations

MRID No. Study Information

487163

- 01 Sargent G. 2012. Product Chemistry: Integral ® F

- 33. Unpublished study prepared by Becker Underwood, Inc., 54 pages.

487163

- 02 Mileson B. 2012. Response to Tier I Microbial Pesticide Data Requirements for Integral ®
F

- 33. Unpublished study prepared by Technology Sciences Group, Inc., 28 pages.

487164

- 01 Sargent G. 2011

. Product Chemistry for Bacillus pumilus strain BU F

- 33 Technical

. U npublished study prepared by Becker Underwood, Inc., 52 pages.

487164

- 02 Monds K. 2011. Bacillus pumilus strain BU F

- 33

– Acute Injection Toxicity/Pathogenicity Study in Rats (Amended Final Report). Project Number: 13682/10. Unpublished study prepared b y Stillmeadow, Inc., 28 pages.

487164

- 03 Kuhn J. 2010. Bacillus pumilus strain BU F

- 33

– Acute Oral Toxicity Study (UDP) in Rats (Amended Final Report). Project Number: 13683/10. Unpublished study prepared by Stillmeadow, Inc., 13 pages.

487164

- 04 Ku hn J. 2010. Bacillus pumilus strain BU F

- 33

– Acute Dermal Toxicity Study in Rats (Amended Final Report). Project Number: 13684/10. Unpublished study prepared by Stillmeadow, Inc., 14 pages.

487164

- 05 Doig A. 2010. Bacillus pumilus strain BU F

- 33

– Acut e Inhalation Toxicity Study in Rats (Amended Final Report). Project Number: 13685/10. Unpublished study prepared by Stillmeadow, Inc., 19 pages.

487164

- 06 Kuhn J. 2010. Bacillus pumilus strain BU F

- 33

– Acute Eye Irritation Study in Rabbits (Amended Fin al Report). Project Number: 13686/10. Unpublished study prepared by Stillmeadow, Inc., 18 pages.Bacillus pumilus strain BU F

- 33 Page 22 of 32 Biopesticides Registration Action Docum ent

MRID No. Study Information

487164

- 07 Kuhn J. 2010. Bacillus pumilus strain BU F

- 33

– Acute Dermal Irritation Study in Rabbits (Amended Final Report). Project Number: 13687/10. Unpubli shed study prepared by Stillmeadow, Inc., 13 pages.

487164

- 08 Younger C. 2010. Bacillus pumilus strain BU F

- 33

– Acute Oral Toxicity Study in Bobwhite Quail (Final Report). Project Number: 13728/10. Unpublished study prepared by Stillmeadow, Inc., 19 pa ges.

487164

- 09 Monds K. 2011. Bacillus pumilus strain BU F

- 33

– Intravenous Toxicity/Pathogenicity Study in Rats (Final Report). Project Number: 15356/11. Unpublished study prepared by Stillmeadow, Inc., 31 pages.

487164

- 10 Mileson B. 2011. Response to Tier I Microbial Pesticide Data Requirements for Bacillus pumilus strain BU F

- 33. Unpublished study prepared by Technology Sciences Group, Inc., 102 pages.

487991

- 01 Sargent G. 2011. pH of Bacillus pumilus strain BU F

- 33 Technical. Unpublished study prepared by Becker Underwood, Inc., 4 pages.

490285

- 01 Sargent G. 2012. Product Chemistry for Bacillus pumilus strain BU F

- 33 Technical: Amendment to MRID No. 487164

- 01. Unpublished study prepared by Becker Underwood, Inc., 20 pages.

490286

- 01 Sargen t G. 2012. Product Chemistry for Integral ® F

- 33: Amendment to MRID No. 487163

- 01. Unpublished study prepared by Becker Underwood, Inc., 54 pages.

490302

- 01 Osprey Biotechnics, Inc. 2013. Description of Starting Materials, Production, and Formulation Pro cess ( Bacillus pumilus strain BU F

- 33 Technical): Amendment to Product Chemistry Data for MRID No. 487164

- 01. Unpublished study prepared by Osprey Biotechnics, Inc., 13 pages.

490893

- 01 Brattan K. 2013. Description of Starting Materials, Production, and Formulation Process ( Bacillus pumilus strain BU F

- 33 Technical): Amendment to Product Chemistry Data for MRID No. 487164

- 01. Unpublished study prepared by Osprey Biotechnics, Inc., 16 pages.Bacillus pumilus strain BU F

- 33 Page 23 of 32 Biopesticides Registration Action Docum ent

B. Environmental Protection Agency Risk Assessment Memorand a

U.S. EPA. 201 3a. Review of the Registrant’s Response to Deficiencies Found in Their Submission for Section 3 Registration of a New TGAI Bacillus pumilus strain BU F

- 33 (EPA Reg. No. 71840

- RE) . Memorandum from I.S. Barsoum, Ph.D. through J.L. Kough, Ph.D . to J. Kausch

, dated February 22 , 2013

.
U.S. EPA. 2013b. Review of the Registrant’s Response to the Deficiency Found in Their Submission for Section 3 Registration of a New EP Integral F

- 33 (EPA Reg. No. 71840

- RG) Containing the TGAI Bacillus pumilus str ain BU F

- 33 (EPA Reg. No.71840

- RE). Memorandum from I.B. Barsoum, Ph.D. through J.L. Kough, Ph.D. to J. Kausch

, dated February 22, 2013.

U.S. EPA. 2013c. Review of Data Waiver Rationale to Satisfy the Microbial Pesticide Data Requirement for Nontarget P lant Testing, Tier I [OCSPP 885.4300] Nontarget Plants in Support for Section 3 Registration of Bacillus pumilus strain BU F

- 33 Technical Grade Active Ingredient [EPA Reg. No. 71840

- RE] and INTEGRAL ® F

- 33 (End

- use Product) [EPA Reg. No. 71840

- RG]. Memorand um from A. Waggoner to J. Kausch

, dated March 30, 2013.

U.S. EPA. 2013d. Environmental Risk Assessment of Bacillus pumilus strain BU F

- 33 for FIFRA Section 3 Registrations of Bacillus pumilus strain BU F

- 33 Technical (EPA File Symbol 71840

- RE) and INTEGRA L ® F

- 33 (EPA File Symbol 71840

- RG) for Use as a Commercial Seed Treatment. Memorandum from S. Borges to J. Kausch

, dated May 7, 2013.

U.S. EPA. 2012a. Review of Product Chemistry and Acute Toxicity Studies for Section 3 Registration of a New TGAI Bacillu s pumilus strain BU F

- 33 (EPA Reg. No. 71840

- RE). Memorandum from I.S. Barsoum, Ph.D. through J.L. Kough, Ph.D. to J. Kausch

, dated November 6, 2012.

U.S. EPA. 2012b. Review of Product Chemistry and Waiver Request for Acute Toxicity Studies for Section 3 Registration of a New EP Integral F

- 33 (EPA Reg. No. 71840

- RG) Containing a New TGAI Bacillus pumilus strain BU F

- 33 (EPA Reg. No. 71840

- RE). Memorandum from I.S. Barsoum, Ph.D. through J.L. Kough, Ph.D. to J. Kausch

, dated November 6, 2012.

C. Other Re ferences

Cottyn B, Regalado B, Lanoot B, De Cleene M, Mew TW, Swings J. 2001. Bacterial populations associated with rice seed in the tropical environment. Phytopathology 91(3):282

– 292.

Garbeva P, van Veen JA, van Elsas JD. 2003. Predominant Bacillus spp . in agricultural soil under different management regimes detected via PCR

- DGGE. Microbial Ecology 45:302

– 316.

Halverson LJ, Jones TM, Firestone MK. 2000. Release of intracellular solutes by four soil bacteria exposed to dilution stress. Soil Science So ciety of America Journal 64:1630

– 1637.

Ivanova EP, Vysotskii MV, Svetashev VI, Nedashkovskaya OI, Gorshkova NM, Mikhailov VV, Yumoto N, Shigeri Y, Taguchi T, Yoshikawa S. 1999. Characterization of Bacillus strains of marine origin. International Microbi ology 2:267

– 271.Bacillus pumilus strain BU F

- 33 Page 24 of 32 Biopesticides Registration Action Docum ent
Juhnke ME, Mathre DE, Sands DC. 1987. Identification and characterization of rhizosphere

- competent bacteria of wheat. Applied and Environmental Microbiology 53(12):2793

– 2799.

Kornegay ET, Risley C R. 1996. Nutrient digestibilities of a c orn

- soybean meal diet as influenced by Bacillus products fed to finishi ng swine. Journal of Animal Science 74: 799

– 805.

Merina M, Lipton AP, Wesley SG. 2011. Isolation, characterization and growth response of biofilm forming bacteria Bacillus pumilus from the sea grass, Halodule pinifolia , off Kanyakumari coast. Indian Journal of Marine Sciences 40(3):443

– 448.

Oguntoyinbo FA. 2007. Monitoring of marine Bacillus diversity among the bacteria community of sea
water. African Journal of Biotechnology 6 (2):163

– 166.

Puente ME, Bashan Y, Li CY, Lebsky VK. 2004. Microbial populations and activities in the rhizoplane of rock

- weathering desert plants. I. Root colonization and weathering of igneous rocks. Plant Biology 6(2004):629

– 642.

U.S. EPA. 2012 c

. Bacillus pumilus Preliminary Work Plan and Summary Document (signed December 18, 2012) (available as a “Supporting Document” within docket ID number EPA

- HQ

- OPP

- 2012

- 0857 at http://www.regulations.gov ).

U.S. EP A. 2012d. Chapter 7 of the Label Review Manual (Precautionary Statements). Available from http://www.epa.gov/oppfead1/labeling/lrm/chap

- 07.pdf

.

U.S. EPA. 2012 e . Chapter 8 of the Label Rev iew Manual (Environmental Hazards). Available from http://www.epa.gov/oppfead1/labeling/lrm/chap

- 08.pdf

.

U.S. EPA. 1996 a . Microbial Pesticide Test Guidelines

– Background for Residue Anal ysis of Microbial Pest Control Agents (OPPTS 885.2000). Available from http://www.epa.gov/ocspp/pubs/frs/publications/Test_Guidelines/series885.htm

.

U.S. EPA. 199 6b . Microbial Pesticide Test Guidelines

– Acute Injection Toxicity/Pathogenicity (OPPTS 885.3200). Available from http://www.epa.gov/ocspp/pubs/frs/publications/Tes t_Guidelines/series885.htm

.
World Health Organization. 2011. Guidelines for Drinking

- water Quality (Fourth Edition). Available from http://whqlibdoc.who.int/publications/2011/ 9789241548151_eng.pdf

.
Yasinok AE, Biran S, Kocabas A, Bakir U. 2010. Xylanase from a soil isolate, Bacillus pumilus : gene isolation, enzyme production, purification, characterization and one

- step separation by aqueous

two

- phase system. World Journal o f Microbiology and Biotechnology 26:1641

– 1652.Bacillus pumilus strain BU F

- 33 Page 25 of 32 Biopesticides Registration Action Docum ent

A PPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS (40 C

. F

. R

. PART 158

– SUBPART V )

TABLE 1. Product Analysis Data Requirements for the Technical Grade of the Active Ingredient (TGAI) , Bacillus pumil us strain BU F

- 33, and Its Manufacturing

- Use Pesticide Product (MP) , Bacillus pumilus strain BU F

- 33 Technical (40 C

. F

. R

. § 158.2 120 )
Harmonized

G uideline N umber Data Requirement Results MRID No. TGAI MP

885.1100 Product Identity N/A Submitt ed dat a fulfill the requirement for product identity. Bacillus pumilus strain BU F

- 33 Technical contains 95.0% by weight Bacillus pumilus strain BU F

- 33 (contains not less than 5.0 x 10 11 cfu per gram ). 487164

- 01

885.1200 Manufacturing Process Submitted data fulfill the requiremen t for manufacturing process

. 487164

- 01 490285

- 01 490302

- 01 490893

- 01

885.1250 Deposition of a Sample in a Nationally Recognized Culture Collection Bacillus pumilus strain BU F

- 33 is on deposit with the Agricultural Research Service Culture Collection (also known as the Northern Regional Research Laboratory) in Peoria, Illinois under Accession Number B

- 50185. N/A

487164

- 01

885.1300 Discussion of Formation of Unintentional Ingredients Submitted data fulfill the requiremen t for discus sion of formation of unintentional ingredients.

885.1400 Analysis of Samples Submitted data fulfill the requirement for analysis of samples.
487164

- 01 490285

- 01 885.1500 Certification of Limits N/A Limits listed on the confidential statement of formula are adequate/acceptable.Bacillus pumilus strain BU F

- 33 Page 26 of 32 Biopesticides Registration Action Docum ent

TABLE 2

. Product Analysis Data Requirements for the End

- Use Pesticide Product ( E P), Integral ® F

- 33 (40 C

. F

. R

. § 158.2120)
Harmonized

Guideline Number Data Requirement Results MRID No.

885.1100 Product Identit y Submitted data fulfill the requirement for product identity. Integral ® F

- 33 contains 5.00 % by weight Bacillus pumilus strain BU F

- 33 (contains not less than 2.2 x 10 10 cfu per milliliter )

. 487163

- 01 490286

- 01
885.1200 Manufacturing Process Submitted data fulfill the requirement for manufacturing process.

885.1250 Deposition of a Sample in a Nationally Recognized Culture Collection N/A N/A

885.1300 Discussion of Formation of Unintentional Ingredients Submitted data fulfill the requiremen t for discussion of formation of unintentional ingredients. 487163

- 01 490286

- 01 885.1400 Analysis of Samples Submitted data fulfill the requiremen t for analysis of samples.

885.1500 Certification of Limits Limits listed on the confidential statement of formula are adeq uate/acceptable.

TABLE 3. Physical and Chemical Characteristics for the Technical Grade of the Active Ingredient (TGAI), Bacillus pumilus strain BU F

- 33, and Its Manufacturing

- Use Pesticide Product (MP), Bacillus pumilus strain BU F

- 33 Technical (40 C.F .R. § 158.2120)
Harmonized Guideline Number Data Requirement Results MRID No. TGAI MP 830.6302 Color Beige

- white

N/A

487164

- 01 490285

- 01 830.6303 Physical State Powder 830.6304 Odor Musty

830.6313 Stability to Normal and Elevated Temperatures, Metals, and Metal Ions Stable at normal and elevated (warehouse) temperatures and expected to be stable when in contact with metals and metal ions. 830.6317 Storage Stability Stable for 12 months at refrigerated and room temperatures.

830.6319 Miscib ility

N/A Not required because Bacillus pumilus strain BU F

33 Technical is not an emulsifiable liquid form of a microbial pesticide (refer to test note #3 of 40 C.F.R. § 158.2120(d)).

830.6320 Corrosion Characteristics Not corrosive to plastic for 12 m onths at refrigerated and room temperatures. 830.7000 pH 4.5

– 6.0 (1% dispersion at 20°C) N/A 487991

- 01 490285

- 01

830.7100 Viscosity N/A Not required because Bacillus pumilus strain BU F

33 Technical is not a liquid form of microbial pesticide (refer to test note #5 of 40 C.F.R. § 158.2120(d)). 487164

- 01 490285

- 01

830.7300 Density/Relative Density/Bulk Density (Specific Gravity) 28.7 +/

- 2.0 lb/ft 3 N/ABacillus pumilus strain BU F

- 33 Page 27 of 32 Biopesticides Registration Action Docum ent

TABLE 4. Physical and Chemical Characteristics for the End

- Use Pesticide Product (EP), Integral ® F

- 33 (40 C

. F

. R

. § 158.2120) Harmonized

Guideline Number Data Requirement Results MRID No.

830.6302 1 Color Tan with solids dispersed 487163

- 01 490286

- 01 830.6303 1 Physical State Liquid 830.6304 1 Odor Musty

830.6313 Stability to Normal and Elevated Te mperatures, Metals, and Metal Ions N/A N/A

830.6317 Storage Stability Stable after 12 months of storage.

487163

- 01 490286

- 01 830.6319 Miscibility Solid

- liquid suspension

830.6320 Corrosion Characteristics Study is in progress. As a term of the Integral ® F

- 33 registration, the EPA is requiring that the results of a

one - year corrosion characteristics study be submitted within one year of registration. 830.7000 1 pH 5

– 7 830.7100 Viscosity 240 cPs +/

- 50 cPs

830.7300 1 Density/Relative Density/Bulk Den sity

(Specific Gravity) 9.73 lb/gal

1 According to 40 C

. F

. R

. § 158.2120, these data are only required for the technical grade of the active ingredient. Since Becker Underwood, Inc.

,

included this information with its application for Integral ® F

- 33 , i t is summarized appropriately in this table.Bacillus pumilus strain BU F

- 33 Page 28 of 32 Biopesticides Registration Action Docum ent

T ABLE 5

. Toxicology Data Requirements for the Technical Grade of the Active Ingredient (TGAI) , Bacillus pumilus strain BU F

- 33, and Its Manufacturing

- Use Pesticide Product (MP), Ba cillus pumilus strain BU F

- 33 Technical (40 C

. F

. R

. § 158.2140) Harmonized

Guideline N umber Data Requirement Results MRID N o. TGAI MP
Tier I

885.3050 Acute Oral Toxicity/Pathogenicity Testing waived based on the applicant’s scientific rationale, whic h cited to MRID Nos. 487164

- 03 (acute oral toxicity study conducted with the MP) and 487164

- 02/487164

- 09 (acute injection toxicity/pathogenicity studies conducted with the TGAI). Classification: Acceptable

N/A 487164

- 10

885.3150 Acute Pulmonary Toxicity /Pathogenicity Testing waived based on the applicant’s scientific rationale, which cited to MRID Nos. 487164

- 05 (acute inhalation toxicity study conducted with the MP) and 487164

- 02/487164

- 09 (acute injection toxicity/pathogenicity studies conducted with the TGAI). Classification: Acceptable

885.3200 Acute Injection Toxicity/Pathogenicity Not infective o r pathogenic to rats when administered intravenously in a single dose of 6.1 x 10 10
cfu/animal. Classification: Acceptable 487164

- 02

Not toxic, in fective, and/or pathogenic to rats when administered intravenously in a single dose of 5.2 x 10 7 cfu/animal. Classification: Acceptable 487164

- 09

885.3400 Hypersensitivity Incidents The applicant reported that n o hypersensitivity incidents, including im mediate

- type or delayed

- type reactions of humans and domestic animals

, occurred durin g research, development, or testing of the TGAI or MP

. Any future hypersensitivity incidents must be reported to the EPA (refer to test note #3 of 40 C

. F

. R

. § 158.2140(d))

. 487164

- 10

885.3500 Cell Culture Not required because Bacillus pumilus strain BU F

- 33 is not a virus (refer to test note #4 of 40 C

. F

. R

. § 158.2140(d)). N/A N/A

870.1100 Acute Oral Toxicity

N/A Oral LD 50 (female rats) > 5,000 mg/kg Classification: Acce ptable TOXICITY CATEGORY IV 487164

- 03

870.1200 Acute Dermal Toxicity Dermal LD 50 (male rats, female rats, and male and female rats combined) > 5,050 mg/kg Classification: Acceptable TOXICITY CATEGORY I V 487164

- 04

870.1300 Acute Inhalation Toxicity Inha lation LC 50 (male rats, female rats, and male and female rats combined) > 2.22 mg/L Classification: Acceptable TOXICITY CATEGORY IV 487164

- 05Bacillus pumilus strain BU F

- 33 Page 29 of 32 Biopesticides Registration Action Docum ent

T ABLE 5

. Toxicology Data Requirements for the Technical Grade of the Active Ingredient (TGAI) , Bacillus pumilus strain BU F

- 33, and Its Manufacturing

- Use Pesticide Product (MP), Ba cillus pumilus strain BU F

- 33 Technical (40 C

. F

. R

. § 158.2140) Harmonized

Guideline N umber Data Requirement Results MRID N o. TGAI MP

870.2400 Acute Eye Irritation

N/A Bacillus pumilus strain BU F

- 33 Technical

was non

- irritating to the eyes of rab bits. No corneal opacity, iritis, conjunctivitis, or other

signs of ocular irritation were noted at any time point in the study. Classification: Acceptable TOXICITY CATEGORY IV 487164

- 06

870.2500 Primary Dermal Irritation Bacillus pumilus strain BU F

- 33 was a slight irritant to the skin of rabbits. All animals were free of slight erythema at 72 hours. Classification: Acceptable TOXICITY CATEGORY IV 487164

- 07

Tiers II and III Not required for Bacillus pumilus strain BU F

- 33 based on the lack of acute tox icity/pathogenicity attributed to the bacterium in the Tier I studies.Bacillus pumilus strain BU F

- 33 Page 30 of 32 Biopesticides Registration Action Docum ent

TABLE 6

. Toxicology Data Requirements for the End

- Use Pesticide Product (EP ) , Integral ® F

- 33 (40 C

. F

. R

. § 158.2140)
Harmonized

Guideline Number Data Req uirement Results MRID No.

885.3050 Acute Oral Toxicity/Pathogenicity

N/A N/A 885.3150 Acute Pulmonary Toxicity/Pathogenicity

885.3200 Acute Injection Toxicity/Pathogenicity

885.3400 Hypersensitivity Incidents The applicant reported than n o hypersen sitivity incidents, including immediate

- type or delayed

- type reactions of humans and domestic animals

, occurred during research, development, or testing of Bacillus pumilus strain BU F

- 33 (includes Integral ® F

- 33)

. Any future hypersensitivity incidents mus t be reported to the EPA (refer to test note #3 of 40 C

. F

. R

. § 158.2140(d)). 487164

- 10

885.3500 Cell Culture N/A N/A

870.1100 Acute Oral Toxicity Testing w aived based on the applicant’s scientific rationale, which cited
to the results of MRID No. 487164

- 03 (acute oral toxicity study conducted with the MP , see Table 5 ) , and because this formulation contains inert ingredients that are not expected to be of toxicological concern (refer to test note #5 of 40 C.F.R. § 158.2140(d)). Classification: Acceptable T OXICITY CATEGORY IV

487163

- 02 870.1200 Acute Dermal Toxicity Testing w aived based on the applicant’s scientific rationale, which cited
to t he results of MRID No. 487164

- 04 (acute dermal toxicity study conducted with the MP , see Table 5 )

, and because this formulation contains inert ingredients that are not expected to be of toxicological concern (refer to test note #5 of 40 C.F.R. § 158.2140(d)). Classification: Acceptable TOXICITY CATEGORY IV

870.1300 Acute Inhalation Toxicity Testing w aived based on the applicant’s scientific rationale, which cited
t o the results of MRID No. 487164

- 05 (acute inhalation toxicity study conducted with the MP , see Table 5 )

, and because this formulation contains inert ingredients that are not expected to be of toxicological

c oncern (refer to test note #5 of 40 C.F.R. § 158.2140(d)). Classification: Acceptable TOXICITY CATEGORY IV

870.2400 Acute Eye Irritation Testing w aived based on the applicant’s scientific rationale, which cited to the results of MRID No. 487164

- 06 (acute eye irritation study conducted with the MP, see Table 5) , and because this formulation contains inert ingredients that are not expected to be of toxicological concern (refer to test note #5 of 40 C.F.R. § 158.2140(d)). Classification: Acceptable TOXICITY CATEGORY IV

870.2500 Primary Dermal Irritation Testing waived based on the applicant’s scientific rationale, which cited to the results of MRID No. 487164

- 07 (primary dermal irritation study conducted with the MP, see Table 5) , and because this formulati on contains inert ingredients that are not expected to be of toxicological concern (refer to test note #5 of 40 C.F.R. § 158.2140(d)). Classification: Acceptable TOXICITY CATEGORY IVBacillus pumilus strain BU F

- 33 Page 31 of 32 Biopesticides Registration Action Docum ent

TABLE 7

. Nontarget Organism Toxicity and Environmental Fate Data Re quirements for the Technical Grade of the Active Ingredient (TGAI), Bacillus pumilus strain BU F

- 33 (40 C

. F

. R

. § 158.2150) Harmonized Guideline Number Data Requirement Results MRID No.

Tier I

885.4050 Avian Oral Toxicity The oral LD 50 for Bacillus pumil us strain BU F

33 was greater than 5,000 mg/kg body weight or 3.19 x 10 12 cfu/kg body weight per day (1.6 x 10 13 cfu/kg body weight total). Bacillus pumilus strain BU F

- 33 is not toxic or pathogenic to birds. Classification: Acceptable 487164

- 08

885.4100 Avian Inhalation Toxicity/Pathogenicity The scientific rationale provides sufficient information to determine that toxicity/pathogenicity to birds is unlikely

. Exposure is expected to be minimal due to the proposed uses (crops), use pattern, and applicatio n rates, as labeled. Classification: Acceptable

487164

- 10 885.4150 Wild Mammal Toxicity/Pathogenicity Tests required by 40 C.F.R. § 158.2140 are adequate to assess hazard of Bacillus pumilus strain BU F

- 33 to wild mammals. The scientific rationale cites acceptable rat acute injection toxicity/pathogenicity studies (MRID Nos. 487164

- 02 and 487164

- 09) and an acceptable rat acute oral toxicity study (MRID No. 487164

- 03). Based on these data, adverse effects to wild mammals are not expected. Classification: A cceptable

885.4200 Freshwater Fish Toxicity/Pathogenicity
The scientific rationale provides sufficient information to determine that toxicity/pathogenicity to these nontarget taxa is unlikely

. Exposure is expected to be minimal due to the proposed uses ( crops), use pattern, and application rates, as labeled. Classification: Acceptable 885.4240 Freshwater Invertebrate Toxicity/Pathogenicity

885.4280 Estuarine/Marine Fish and Invertebrate Testing

885.4300 Nontarget Plant Testing

885.4340 Nontarg et Insect Testing

885.4380 Honey Bee Testing
Tiers II, III, and IV Not required for Bacillus pumilus strain BU F

- 33 based on the acceptability of the scientific rationale and study provided for Tier I

.Bacillus pumilus strain BU F

- 33 Page 32 of 32 Biopesticides Registration Action Docum ent

APPENDIX B. PESTICIDE PRODUCTS

EP A File Symbol Registration Name Percentage Active Ingredient Formulation Type Use Site(s) Method(s) of Application Application Rate (s) Target Pest

71840

- RE Bacillus pumilus strain BU F

- 33

Technical 95.0 % Technical N/A

71840

- RG Integral ® F

- 33 5.00% End U se

– Liquid Seed of certain crops/plants Standard commercial seed
treatment equipment Small containers 0.01 fl oz

– 10 fl oz of Integral ® F

- 33 per 100 lb seed

Large containers 1 fl oz

– 7.8 gal of Integral ® F

- 33 per

10,000 lb seed Elicits induced systemi c resistance in plants

, thereby enhancing their defense against a spectrum of plant pathogens, including

viruses, fungi, and bacteria.