### 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

June 12 , 2013Bacillus pumilus strain BU F

- 33 Page 2 of 32 Biopesticides Registration Action Document



### TABLE OF CONTENTS

I. EXECUTIVE SUMMARY

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

II. ACTIVE INGREDIENT OV ERVIEW

................................ ................................ ................................ ...................... 6

III. REGULATORY BACKGROUN D

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

A. Applications for Pesticide Product Registration

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

B. Food Tolerance Exemption

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

IV. RISK ASSESSMENT

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

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

................................ ................................ ................................ ............................... 1 7

VI. RISK MANAGEMENT DECI SION

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

VII. ACT IONS REQUIRED OF THE REGISTRANT

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

A. Final Printed Labeling

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

B. Terms of Registration

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

C. Reporting of Adverse Effects and Hypersensitivity Incidents

................................ ................................ ...... 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

APPE NDIX A. MICROBIAL P ESTICIDES DATA REQUI REMENTS

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

APPENDIX B. PESTICI DE PRODUCTS

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

- 33 Page 3 of 32 Biopesticides Registration Action Document



### BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

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

Science Review

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

Regulatory Review

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

- 33 Page 4 of 32 Biopesticides Registration Action Document

I. EXECUTIVE SUMMARY

Background
In January 201 2

, Becker Underwo od , 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 Prote ction 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 bacter ia that are ubiquitous 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 . 2011; 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) hav e been used in registered 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 esta blished the following 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 7661 9 )

.

(2) Bacillus pumilus strain QST 2808 ( 40 C

. F

. R

. § 180.1255 )

– see the Federal Register of November 3, 2 004 ( 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 healt hy cucumber plant at 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, fun g i, and bacteria . This 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 cienti sts reviewed product 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 product s. Overall, such data
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 Document

Product Analysis
The applicant ful filled 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 submi tted a dequate mamm alian 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 , infective, 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 tox icity, 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 testin g of Bacillus pumilus 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 co ncludes 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 strain 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 beca use of (1) the application 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 p lant 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 rem ove microbes or render 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 drinking 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

, th e 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 commerc ial 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 future 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 Document

Nontarget Organisms
The EPA found that s cientific rationale and an acute avian oral to xicity/pathogenicity 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

req uired. The EPA performed 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 Partic ipation
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 mak ing 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, and allowing for this active ingredient’s first food and outdoor uses were subject to a 15

- day comment period. During this comment period, the EPA received no comments on its preliminary decision to register the Bacillus pumilus strain BU F

- 33 pesticide products, Bacillus pumilus strain BU F

- 33 Technical and Integral ®

F

- 33. Ther efore, the EPA maintains that, based upon the risk assessment and information submitted in support of such pesticide products, it is appropriate to issue these registrations. The basis for this
decision can be found in the risk assessment for Bacillus pumi lus strain BU F

- 33 , which is characterized throughout this Biopesticides Registration Action Document ( BRAD )

.

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 registered pesticide products containing this active ingredient.Bacillus pumilus strain BU F

- 33 Page 7 of 3 2 Biopesticides Registration Action Document

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 application s to register a manufacturing

- use pesticide product, Ba cillus 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). The
EPA received no co mments following this publication.

B. Food Tolerance Exemption

Concurrent with its registration application s and under Federal Food, Drug, and Cosmetic Act (FFDCA) section 408(d), Becker Underwood, 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 U nderwood, 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. The EPA r eceived no comments following this publication.

On J une 12 , 201 3

, the EPA established an exemption from the requirement of a tolerance for residues of
B acillus pumilus strain BU F

- 33 in or on all food commodities when applied to elicit induced syste mic resistance in plants and used in accordance with label directions and good agricultu ral practices (40 CFR § 180.13 22 ; 7 8 FR 35147 ).

IV. RISK ASSESSMENT

In the Federal Register s of October 26, 2007 ( 72 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 for 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 usefulness 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 can be useful for risk assessment. The studies may have certa in 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 “ acceptabl e .” 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 risk assessment

process as appropriate. An “unacceptable ” ra ting 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 pesticideBacillus pumilus strain BU F

- 33 Page 8 of 32 Biopesticides Registration Action Document

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 product

- specific information.

B. Human Health Assessment ( 40 C

. F

. R

. § 158.2140 )

1. Toxicity

The applicant 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 ba sed 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 IV(B)(1)(b), as well as additional product

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

.

a. Acute Toxicity/Pathogenicity

– Tier I

Acute Oral Toxicity/Pathogenicity ( Harmonized Guideline 885.3050; Master Record Identificatio n 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 Bacillus pumilus
strain BU F

- 33. An acute oral toxicity study cond ucted 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) sh owed 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 this bacterium would not be toxic, infective, and/or pathogenic t hrough 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 results 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) demonstrated that Bacillus pumilus
strain BU F

- 33 was not toxic (LC 50 gr eater 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 toxic, 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 further testing is not

necessary

.Bacillus pumilus strain BU F

- 33 Page 9 of 32 Biopesticides Registration Action Document

Acute Injection Toxicity/Path ogenicity (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 Sprague

- 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 intraven ous 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 cumulative 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 three 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 females killed on day 21. Enlarged lymph nodes were an additi onal 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 weight in inactive

- and active

- treated animals of
both sexes (ma les: +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 org anism 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. The EPA rated this study as acceptable.

Regarding the toxicity uncertainty mentioned above, i t is kno wn 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 ( 19 96b ) ) . 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 t ested 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 study ).

Acute Injection Toxicity/Pathogenicity (I ntravenous)

– 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 (3Bacillus pumilus strain BU F

- 33 Page 10 of 32 Biopesticides Registration Action Document

animals per sex) taking place on days 0, 14, 21, 28, a nd 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 we ight 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 content samples taken on day 14. The test organi sm 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 42. For the lungs, all values were
less than 10 0 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 appear to be toxic, infective, and/or pathogenic in rats. The EPA rated this study as 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 an d 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

. § 158.2140(d))

.

Cell Culture ( Harmonized Guidel ine 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)).

b. Acute Toxicology and Subchronic Toxicity/Pathogenicity

– Tier II; Reproduc tive 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 FFDCA 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 eff ect 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 requiremen ts 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 sub stance.Bacillus pumilus strain BU F

- 33 Page 11 of 32 Biopesticides Registration Action Document

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
strain 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) EDS P. 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 test 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 EPA to establish an exemption from the r equirement 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 “there is a reasonable certainty that no ha rm will result from aggregate exposure to the pesticide chemical residue, including all anticipated dietary exposures and all other exposures for which there is reliable information.” This includes exposure through drinking water and in residential setting s but does not include occupational exposure. Pursuant to FFDCA section 408( c )(2)(B) , in establishing or maintaining in effect an exemption from the requirement of a tolerance, the EPA must take into account the factors
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 an d 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 particular pesticide's] . . . residues and othe r 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 t he 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 scientific data and other
relevant informatio n 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 o f 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.Bacillus pumilus strain BU F

- 33 Page 12 of 32 Biopesticides Registration Action Document

a. Aggregate Exposure

In examining aggregate exposure, FFDCA section 408 directs the EPA to consider available information concerning exposures from the pesticide re sidue 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 Characteriza tion : 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; Y asinok 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, treated seeds would be bagged, sold to gr owers, 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 clo se 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; therefore, exposure to Bacillus pumilus stra in 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 with any food 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 ).

Drinking Water Exposure and Ri sk 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; Merina et al . 2011; Oguntoyinbo 2007 ; Pu ente 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 treatment that takes place indoors. Comme rcial 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 obtain a National Pollutant Discharge Eli mination 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, for larger container sizes, it is possibl e 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 growers will colonize and grow with the ro ots 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 waters 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 facilities (e.g., pH adjustments and chlorinati on). 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 resistant to these disinfection processes ( World Health Organization 2011 ). In the remote likelihood that Bacillus pumilusBacillus pumilus strain BU F

- 33 Page 13 of 32 Biopesticides Registration Action Document

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 w ater 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 . 2 004; 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 into soil in agricultural settings; diver ted 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 Bacillus pumilus strain BU F

- 33 are anticipa ted, particularly in light of testing that indicates that this bacterium is not toxic (acute inhalation toxicity and acute dermal toxicity tests), is only slightly irritating (primary dermal irritation test), and is not pathogenic or infective (acute injec tion toxicity/pathogenicity test) at high doses ( see section IV(B)(1)(a) and Tables 5 and 6 in Appendix A ).

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

Section 408(b)(2)(D)(v) of FFDCA requi res 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 substances that have a common mechanism of toxi city.”

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 produced by other substances. For the purpose s 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

assoc iated 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 effects of such chemicals, see the EPA’s websi te 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 est ablishment 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 infants and children to pesticide chemical re sidues, 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 additional 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 chi ldren. This additional margin of safety is commonly referred to as the Food QualityBacillus pumilus strain BU F

- 33 Page 14 of 32 Biopesticides Registration Action Document

Protection Act Safety Factor. In applying this provision, the EPA either retains the default value of 10X, or uses a different additional 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 t hat 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 that no additional margin of exposure (safe ty) is necessary.

Moreover , based on the same data and analysis as presented directly above, the EPA conclude s that there is a reasonable certainty that no harm will result to the U.S. population, including infants and children, from aggregate exposure to the residues of Bacillus 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 includes all anticipated dietary exposures and all other e xposures 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 demonstrate toxic, pathogenic, and/or infective potentia l to mammals, including infants and children.

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 pumilus strain BU F

- 33 in commercial seed

treatment setting s 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 future produ cts, 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 human exposure to Bacillus pumilus strain BU F

- 33 in ligh t 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 incident al exposures when Bacillus pumilus strain BU F

- 33 pesticide products are applied to seed in accordance with the EPA’s approved labeling.Bacillus pumilus strain BU F

- 33 Page 15 of 32 Biopesticides Registration Action Document

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 applicant are sufficient to fulfill the nontarget organ ism 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 assessment, based on information provided by the

ap p licant, 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” determination for direct and indirect effects to fed erally 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 sections 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 cas es. 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 profile within the soil to areas around plants gro wing from treated seed. Exposure to Bacillus pumilus strain BU F

- 33 in soil is not expected to be significant.

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

- 08 and 4 87164

- 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 cause
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 1 2 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 inhalatio n 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

no t 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 exposed 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 inf ect) 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 result of the proposed registrations of the Bacillus pu milus strain BU F

- 33 pesticide products

.Bacillus pumilus strain BU F

- 33 Page 16 of 32 Biopesticides Registration Action Document

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 included. In this study, Bacillus pumilus was associated w ith 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 minimize the potential for Bacillus pumilus BU F

- 33 to con tact 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 pathogenic to crop plants (U

. S

. EPA 2013 c ), which may ind icate 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 application 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 nontarget 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 nontarget 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 contact with soil
and water

. Therefore, based on minim al 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 BU
F

- 33 pesticide products.

d. Freshwater Fish (Harmonized Guideline 885.4200), Fre shwater 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
envir onments 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 presents 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 Nontarget Organisms

Based on the expected lack of exp osure 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

pro ducts , containing Bacillus pumilus strain BU F

- 33, as labeled.Bacillus pumilus strain BU F

- 33 Page 17 of 32 Biopesticides Registration Action Document

2. Environmental Fate Data

As t he rationale and data provided are sufficient to fulfill the Tier I nontarget organism data requirements and allow for nontarget organism risk asse ssment f or 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 that no effects are anticipated for nontarget specie s 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 expected. Therefore, a “No Effect”
determination is mad e 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 ac hieve environmental justice

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

— with respect to the development, implementation, and enforcement of environmental laws, regulations, and policies. Fair treatment means that no group of people, including racial, ethnic, or socioeconomic groups, should bear a disproportionate share of the negative environmental consequences resulting from industrial, municipal, and commercial operations or the executi on of federal, state, local, and tribal environmental programs and policies. Meaningful involvement means that (1) potentially affected community residents have an appropriate opportunity to participate in decisions about a proposed activity that will affe ct their environment and/or health; (2) the public’s contribution can influence the regulatory agency’s decision; (3) the concerns of all participants involved will be considered in the decision

- making process; and (4)
the decision

- makers seek out and faci litate 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, during the 15

- day public participation comment period, the EPA s ought information on any groups or segments of the population who, as a result their location, cultural practices, or other factors, may have atypical, unusually high exposure to Bacillus pumilus strain BU F

- 33 , compared to the general population. The EPA received no public comments on this particular matter.

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

.Bacillus pumilus strain BU F

- 33 Page 18 of 32 Biopesticides Registration Action Document

V I

. RISK MANAGEMENT DECISION

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

(1) Its composition is such as to warrant the proposed claims 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 unreasonable adverse effects on the environment; AND (4) When used in accordance with widespread and commonly recognized practice, it will not generally cause unreasonable adverse effects on the
environment.

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

- known to elic it 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 pro ducts 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 docume nt. 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 resistanc e 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 when used according to label instructions. Therefore, EPA believe s 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) for the labeled uses.Bacillus pumilus strain BU F

- 33 Page 19 of 32 Biopesticides Registration Action Document

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 labeling to the EPA

.

B. Terms of Registration

T h e EPA is not requiring submission of additional data as terms of the Bacillus pumilus strain BU F

- 33 Technical and Integral ® F

- 33 registrations.

C. Reporting of Adverse Effects and Hypersensitivity Incidents

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

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 Document

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 Cosmetic Act
FI FRA 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 expected 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 cause 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 manufacturi ng

- 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 Pesticide P rograms 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 Document

I X. BIBLIOGRAPHY

A. Studies Submitted to Support the Ba cillus 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

. Unpublished study p repared 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 by 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 Kuhn J. 2010. Bacill us 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

– Acute Inhalation Toxic ity 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 Final Report). Projec t Number: 13686/10. Unpublished study prepared by Stillmeadow, Inc., 18 pages.Bacillus pumilus strain BU F

- 33 Page 22 of 32 Biopesticides Registration Action Document

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. Unpublished study prepare d 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 pages.

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 Microbia l 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 Sargent 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 Process ( Bacillus pum ilus 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 Proce ss ( 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 Document

B. Environmental Protection Agency Risk Assessment Memoranda

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

, da ted 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 strain BU F

- 33 (EPA R eg. 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 Plant 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]. Memorandum from A. Waggone r 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 INTEGRAL ® 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 Bacillus pumilus strain B U 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 References

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 Society of America J ournal 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 Microbiology 2:267

– 271.Bacillus pumilus strain BU F

- 33 Page 24 of 32 Biopesticides Registration Action Document

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 corn

- soybean meal d iet 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.

Pu ente 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 pumilu s 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. EPA. 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 Review Manual (Enviro nmental 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 Analysis 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. 1996b . Microbial Pest icide Test Guidelines

– Acute Injection Toxicity/Pathogenicity (OPPTS 885.3200). Available from http://www.epa.gov/ocspp/pubs/frs/publications/Test_Guidelines/serie s885.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 of Microbiology and Biotechnology 26:1641

– 1652.Bacillus pumilus strain BU F

- 33 Page 25 of 32 Biopesticides Registration Action Document

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 pumilus 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 data fulfill the requ irement 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 requi remen 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 discussion 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/accep table.

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 Identity Submitted data fulfill the requir ement 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 manufa cturing 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 ingr edients. 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 adequate/acceptable.Bacillus pumilus strain BU F

- 33 Page 26 of 32 Biopesticides Registration Action Document

TABLE 3. Physic al 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 Guideli ne 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 n ormal 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 (4

– 5°C) and room (21

– 25°C) temperatures.

830.6319 Miscibility

N/A Not req uired 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 months at refriger ated (4

– 5°C) and room (21

– 25°C) 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 Document

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 Temperatures, Metals, and Metal Ions N/A N/A

830.6317 Storage Stability Stable for 12 months at refrigerated (4

– 5°C) and room (21

– 25°C) temperatures

.

487163

- 01
490286

- 01 830.6319 Miscibility Solid

- liquid suspension

830.6320 Corrosion Chara cteristics Not corrosive to plastic containers for 12 months at refrigerated (4

– 5°C) and room (21

– 25°C) temperatures.
830.7000 1 pH 5

– 7
830.7100 Viscosity 240 cPs +/

- 50 cPs

830.7300 1 Density/Relative Density/Bulk Density

(Specific Gravity) 9.73 lb/ga l

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 , it is summarized appropriately in t his table.Bacillus pumilus strain BU F

- 33 Page 28 of 32 Biopesticides Registration Action Document

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), Bacillus pumilus strain BU F

- 33 Tech nical (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, which cited
to MRID Nos. 487164

- 03 (ac ute 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 base d 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: Accep table

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, infective, 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 immediate

- type or delayed

- type react ions 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 Cultur e 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: Acceptable 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 Inhalation LC 50 (male rats, female rat s, 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 Document

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), Bacillus pumilus strain BU F

- 33 Tech nical (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 rabbits. 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 toxicity/pathogenicity attributed to the bacterium in the Tier I studies.Bacillus pumilus strain BU F

- 33 Page 30 of 32 Biopesticides Registration Action Document

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 Requirement 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 hypersensitivity incidents, including immediate

- type o r 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 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 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 t he 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

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 ar e 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 concern (refer to test note #5 of 40 C.F.R. § 1 58.2140(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, s ee 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
Irritati on 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 formulation contains inert ingredients that are not exp ected 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 Document

TABLE 7

. Nontarget Organism Toxicity and Environmental Fate Data Requirements 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 pumilus strain BU F

- 33 was greater than 5,000 mg/kg b ody 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 scie ntific 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 application 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/pathogen icity 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: Acceptable

885.4200 Freshwater Fish Toxicity/Pa thogenicity

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 l abeled. Classification: Acceptable 885.4240 Freshwater Invertebrate Toxicity/Pathogenicity

885.4280 Estuarine/Marine Fish and Invertebrate Testing

885.4300 Nontarget Plant Testing

885.4340 Nontarget 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 Document

APPENDIX B. PESTICIDE PRODUCTS

EPA Registration

Number Registration
Name Percentage Ac tive Ingredient Formulation Type Use Site(s) Method(s) of Application Application Rate (s) Target Pest

71840

- 12 Bacillus pumilus
strain BU F

- 33

Technical 95.0 % Technical N/A

71840

- 13 Integral ® F

- 33 5.00% End Use

– Liquid Seed of certain crops/plants Stand ard commercial seed treatment equipment Small containers
0.005 fl oz

– 5 fl oz of Integral ® F

- 33 per 50 lb seed

OR

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 i nduced systemic resistance in plants , thereby enhancing their defense against a spectrum of plant pathogens, including viruses, fungi, and bacteria.