BIOPESTICIDES REGISTRATION ACTION DOCUMENT

Bacillus pumilus strain BU F-33

Pesticide Chemical (PC) Code: 007493

U.S. Environmental Protection Agency

Office of Pesticide Programs

Biopesticides and Pollution Prevention Division

DRAFT – May 9, 2013
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Biopesticides Registration Action Document

TABLE OF CONTENTS

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

II. ACTIVE INGREDIENT OVERVIEW .....................................................................................................7

III. REGULATORY BACKGROUND ............................................................................................................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 JUSTICE...............................................................................................................17

VI. RISK MANAGEMENT DECISION........................................................................................................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. MICROBIAL PESTICIDES DATA REQUIREMENTS ........................................................25

APPENDIX B. PESTICIDE PRODUCTS...........................................................................................................32
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Biopesticides Registration Action Document

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 Leader
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Biopesticides Registration Action Document

I. EXECUTIVE SUMMARY

Background

In January 2012, Becker Underwood, Inc., submitted applications for 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), to the U.S. Environmental Protection 

Agency (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 ubiquitous in nature and have 

been identified and isolated from samples of a variety of media, including soil, water, aquatic 

invertebrates, 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 2012c; 

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 registered 

pesticide products to suppress plant pathogens such as Pythium ultimum, Fusarium oxysporum, 

Rhizoctonia solani, and Alternaria species (U.S. EPA 2012c). In conjunction with some of these 

registered pesticide products, the EPA established 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 76619).

(2) Bacillus pumilus 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 March 30, 

2012 (77 FR 19109).

Bacillus pumilus strain BU F-33 was isolated from inside the stem of a healthy cucumber plant at an 

Auburn University facility outside Tallassee, 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, fungi, and bacteria. This increased protection results in more vigorous root 

and shoot systems in treated plants and leads to more uniform 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 scientists 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 products. 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).
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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 adequate mammalian toxicology data and information to support the Bacillus 

pumilus strain BU F-33 pesticide products. An acute injection toxicity/pathogenicity (intravenous) study

showed that, 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 

reported 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 research, development, or testing of Bacillus pumilus strain 

BU F-33. In light 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 strain BU F-33 as an active 

ingredient in pesticide products. Although exposure to Bacillus pumilus strain BU F-33 through

consumed food and drinking water may be possible, the EPA anticipates these types of exposures to be 

negligible because of (1) the application method of 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 render them nonviable (U.S. EPA 1996a); 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 data 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 future products,

the EPA may require additional PPE (e.g., protective eyewear), other than the standard described above, 

on a product-specific basis. 
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Nontarget Organisms

The EPA found that scientific rationale and an acute avian oral toxicity/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 

required. The EPA performed an environmental risk assessment, based on information provided by the 

applicant, 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 federally listed threatened and endangered species 

and their designated critical habitats resulting from these same proposed pesticidal uses. 

Public Participation

On October 1, 2009, the EPA announced a policy to provide a more meaningful opportunity for the 

public to participate in major registration decisions before they occur. According to this policy, the EPA 

provides a public comment period prior to making a registration decision for the following types of 

applications: new active ingredients; first food uses; first outdoor uses; first residential uses; or any other 

registration actions for which the EPA believes there may be significant public interest.

 

Consistent with the policy of making registration actions more transparent, the proposed pesticide 

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 registration of these pesticide 

products would result in the first food and outdoor uses 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-2012-0265. The following documents are available for comment in EPA-HQ-OPP2012-0265: (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 

manufacturing-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 Symbol:

71840-RG). While a final decision on registration is contingent upon review 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. 
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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: 007493

Type of Pesticide: Microbial Pesticide – Plant Regulator

See Appendix B for specific information (e.g.,

use sites, application rates, methods of application, 

formulation types, and target pests) regarding

the proposed pesticide products containing this 

active ingredient.

III. REGULATORY BACKGROUND

A. Applications for Pesticide Product Registration

In January 2012, Becker Underwood, Inc. (address: 801 Dayton Avenue; P.O. Box 667; Ames, Iowa 

50010), submitted applications 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 applications to register pesticide products containing a new active ingredient 

and opened a 30-day public comment period pursuant to the provisions of FIFRA section 3(c)(4). No 

comments were received following this publication. 

B. Food Tolerance Exemption

Concurrent with its registration applications 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 (77 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 comments were received following this publication. 
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Biopesticides Registration Action Document

IV. RISK ASSESSMENT

In the Federal Registers of October 26, 2007 (72 FR 60988) and August 30, 2012 (77 FR 52610), the 

EPA issued Final Rules 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 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 the study is not required by 40 

C.F.R. Part 158. Both “acceptable” and “supplemental” studies may be used in the risk assessment 

process as appropriate. An “unacceptable” rating indicates that new data must be submitted.

For the acute toxicity data requirements, Toxicity Categories are assigned based on the 

hazard(s) identified from studies and/or other information 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 all 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 all 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 

IV(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.
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a. Acute Toxicity/Pathogenicity – Tier I

Acute Oral Toxicity/Pathogenicity (Harmonized Guideline 885.3050; Master Record Identification 

Number (MRID No.) 487164-10): Upon consideration of results of other definitive toxicological 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 conducted on female rats (MRID No. 487164-03) 

demonstrated that Bacillus pumilus strain BU F-33 was not toxic (LD50 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 

this 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 Guideline 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 (LC50 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 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.

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 Sprague-Dawley rats were injected intravenously with Bacillus pumilus strain BU F33 (6.38 x 1011 viable spores per gram) in sterile phosphate buffered saline at a dose of 6.1 x 1010

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) taking 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. Inactive-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 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 increased absolute mesenteric lymph node weight in inactive- and active-treated animals of 
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Biopesticides Registration Action Document

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 1010

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, it 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 1010 vs. 1 x 107 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 107 cfu/animal) in 

the range recommended by U.S. EPA (1996b) was tested in MRID No. 487164-09. This particular test 

demonstrated 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 (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 1011 viable spores per gram) in sterile phosphate buffered saline at a dose of 5.2 x 107

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 content 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 42. 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 107

cfu/animal does not appear 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 no 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. § 158.2140(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)).
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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 FFDCA section 408(p), the EPA has developed the Endocrine 

Disruptor Screening Program (EDSP) and has begun to implement the screening program that is to be 

used to test all pesticides in order to determine whether certain substances (including pesticide active and

other ingredients) may have an effect in humans or wildlife similar to an effect produced by a “naturally 

occurring estrogen, or other such endocrine effects as the Administrator may designate.” 

FFDCA section 408(p)(4) authorizes the Administrator, by order, to exempt from the requirements of 

the Estrogenic Substances Screening Program a biologic substance or other 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

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 that 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, pursuant 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 test guidelines and the Tier 1 screening battery, please visit our website 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 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 “there is a 

reasonable certainty that no harm will result from aggregate exposure to the pesticide chemical residue, 

including all anticipated dietary exposures and all other exposures for which there is reliable

information.” This includes exposure through drinking water and in residential settings but does not 
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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 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 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 particular 

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 scientific 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 

Tables 5 and 6 in Appendix A, the 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

concerning 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, treated 

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; therefore, 

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 

1996a). 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 
Bacillus pumilus strain BU F-33 Page 13 of 32

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applied in accordance with label directions and good agricultural practices (see section IV(B)(1)(a) and 

Tables 5 and 6 in Appendix 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; Merina 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 

treatment 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 obtain 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, for 

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 growers 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 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 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 resistant 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 Tables 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 into 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 Bacillus 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 

not 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).
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b. Cumulative Effects from Substances with a Common Mechanism of Toxicity

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

revoke a tolerance, the EPA consider “available information concerning the cumulative effects of [a 

particular pesticide’s] . . . residues and other substances 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 

produced 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 effects 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 infants and children to 

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

substances with a common mechanism of toxicity. In addition, FFDCA section 408(b)(2)(C) provides 

that 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 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 additional or no safety factor when reliable data are available to support a different 

additional or no safety factor. 

Based on the acute toxicity and 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 that 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, 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 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 demonstrate toxic, pathogenic, 

and/or infective potential to mammals, including infants and children.
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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 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. 

4. Human Health Risk Characterization

The EPA considered human 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 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 performed an environmental risk assessment, based on information provided by the 

applicant, 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 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 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 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 profile 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. 
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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 cause 

adverse effects in these animals. An acute avian toxicity study with Northern bobwhite (Colinus 

virginianus, MRID No. 487164-08) showed no toxicity or pathogenicity due to Bacillus pumilus strain 

BU F-33 (concentration of 6.38 x 1011 cfu/g), when orally administered at 5,000 mg/kg body weight

(3.19 x 1012 cfu/kg body weight) daily for five consecutive days (1.6 x 1013 cfu/kg body weight total). 

Scientific rationale submitted to satisfy the avian inhalation toxicity/pathogenicity study also highlighted 

theinfluence 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 1010 cfu/animal (MRID No. 487164-02) and was 

not infective, pathogenic, and/or toxic to rats when injected at 5.2 x 107 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. This study indicated that the Bacillus pumilus did not colonize 

(or infect) the digestive tracts 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 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 with 

rice seed was also included. 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 minimize 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 pathogenic to crop plants (U.S. EPA 2013c), which may indicate concern for crop 

plants treated (as seed) with Bacillus pumilus strain BU F-33. Based on this review, Becker Underwood, 

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 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 BU 
Bacillus pumilus strain BU F-33 Page 17 of 32

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F-33 pesticide products. 

d. Freshwater 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 

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 exposure and data demonstrating no effects, as discussed above, 

unreasonable adverse effects to nontarget organisms are not anticipated as a result of the proposed 

registrations of the Bacillus pumilus strain BU F-33 Technical and INTEGRAL® F-33 pesticide 

products, containing Bacillus pumilus strain BU F-33, as labeled. 

2. Environmental Fate Data 

As the rationale and data provided are sufficient to fulfill the Tier I nontarget organism data 

requirements and allow for nontarget organism risk assessment 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 that no effects are anticipated for nontarget species exposed to Bacillus 

pumilus strain BU F-33 as a result of the proposed labeled uses, effects to federally listed threatened and 

endangered species and their designated critical habitats are also not expected. 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 uses 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, municipal, 

and commercial operations or the execution of federal, state, local, and tribal environmental programs 

and policies. Meaningful involvement means that (1) potentially affected community residents have an 

appropriate opportunity to participate in decisions about a proposed activity that will affect their 

environment and/or health; (2) the public’s contribution can influence the regulatory agency’s decision; 
Bacillus pumilus strain BU F-33 Page 18 of 32

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(3) the concerns of all participants involved will be considered in the decision-making process; and (4) 

the decision-makers seek out and facilitate the involvement of those potentially affected. The EPA has 

this goal for all communities and persons across the United States. 

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

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

atypical, unusually high exposure to Bacillus 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 website at 

http://www.epa.gov/compliance/environmentaljustice/index.html.

VI. 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 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 labels, and such pesticide products are not expected to cause unreasonable adverse 

effects on the environment when used according to their respective label instructions. Criterion 2 is 

satisfied by the current product labels, 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 likely 

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 when 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) for the labeled uses. 
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VII. 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

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 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 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).
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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 Environmental Protection Agency 

FFDCA Federal Food, Drug, and Cosmetic Act

FIFRA Federal Insecticide, Fungicide, and Rodenticide Act

fl oz fluid ounce

FR Federal Register

gal gallon

lb pound

lb/ft3 pounds per cubic foot

LC50 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).

LD50 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 milligrams per kilogram

mg/L milligrams per liter

MP manufacturing-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 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 States
Bacillus pumilus strain BU F-33 Page 21 of 32

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IX. BIBLIOGRAPHY

A. Studies Submitted to Support 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.

Unpublished 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 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. 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 – Acute 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 Final Report). Project Number: 13686/10. Unpublished study prepared by 

Stillmeadow, Inc., 18 pages.
Bacillus pumilus strain BU F-33 Page 22 of 32

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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 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 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 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 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 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.
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B. Environmental Protection Agency Risk Assessment Memoranda

U.S. EPA. 2013a. 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 strain 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 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. 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 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 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 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 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 Microbiology 2:267–271.
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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 CR. 1996. Nutrient digestibilities of a corn-soybean meal diet as influenced by

Bacillus products fed to finishing 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. 2012c. 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. 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. 2012e. Chapter 8 of the Label Review Manual (Environmental Hazards). Available from 

http://www.epa.gov/oppfead1/labeling/lrm/chap-08.pdf.

U.S. EPA. 1996a. 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 Pesticide Test Guidelines – Acute Injection Toxicity/Pathogenicity (OPPTS 

885.3200). Available from 

http://www.epa.gov/ocspp/pubs/frs/publications/Test_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 aqueoustwo-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

APPENDIX 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. § 158.2120)

Harmonized 

Guideline 

Number 

Data Requirement

Results

MRID No.

TGAI MP

885.1100 Product Identity N/A

Submitted data 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 

1011 cfu per gram).

487164-01

885.1200 Manufacturing Process Submitted data fulfill the requirement 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 requirement for discussion of formation of 

unintentional ingredients.

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

487164-01

885.1500 Certification of Limits N/A 490285-01

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 2. Product Analysis Data Requirements for the End-Use Pesticide Product (EP), 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 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 1010 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 requirement for discussion of formation of 

unintentional ingredients. 487163-01

885.1400 Analysis of Samples Submitted data fulfill the requirement for analysis of samples. 490286-01

885.1500 Certification of Limits Limits listed on the confidential statement of formula are 

adequate/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 Miscibility

N/A

Not required because 

Bacillus pumilus strain BU F33 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 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 F33 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/ft3 N/A
Bacillus 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.

830.63021 Color Tan with solids dispersed 487163-01

490286-01 830.63031 Physical State Liquid

830.63041 Odor Musty

830.6313

Stability to Normal and 

Elevated Temperatures, 

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.

830.70001 pH 5–7

830.7100 Viscosity 240 cPs +/- 50 cPs

830.73001

Density/Relative 

Density/Bulk Density 

(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, it is summarized appropriately in this table.
Bacillus pumilus strain BU F-33 Page 28 of 32

Biopesticides Registration Action Document

TABLE 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 Technical (40 C.F.R. § 158.2140)

Harmonized

Guideline 

Number

Data Requirement

Results

MRID No.

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 (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 or pathogenic to rats 

when administered intravenously in 

a single dose of 6.1 x 1010

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 107 cfu/animal. 

Classification: Acceptable

487164-09

885.3400 Hypersensitivity

Incidents

The applicant reported that no hypersensitivity incidents, including immediate-type 

or delayed-type reactions of humans and domestic animals, occurred during 

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 LD50 (female rats) > 5,000 mg/kg

Classification: Acceptable

TOXICITY CATEGORY IV

487164-03

870.1200 Acute Dermal 

Toxicity

Dermal LD50 (male rats, female rats, and 

male and female rats combined) > 5,050 

mg/kg

Classification: Acceptable

TOXICITY CATEGORY IV

487164-04

870.1300 Acute Inhalation 

Toxicity

Inhalation LC50 (male rats, female rats, and 

male and female rats combined) > 2.22 mg/L

Classification: Acceptable

TOXICITY CATEGORY IV

487164-05
Bacillus pumilus strain BU F-33 Page 29 of 32

Biopesticides Registration Action Document

TABLE 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 Technical (40 C.F.R. § 158.2140)

Harmonized

Guideline 

Number

Data Requirement

Results

MRID No.

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

885.3150 N/A N/A Acute Pulmonary 

Toxicity/Pathogenicity

885.3200 Acute Injection 

Toxicity/Pathogenicity

885.3400 Hypersensitivity 

Incidents

The applicant reported than no 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 (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 waived 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

TOXICITY CATEGORY IV

487163-02

870.1200 Acute Dermal 

Toxicity

Testing waived based on the applicant’s scientific rationale, which cited 

to the 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 waived based on the applicant’s scientific rationale, which cited 

to 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. § 158.2140(d)).

Classification: Acceptable

TOXICITY CATEGORY IV

870.2400 Acute Eye Irritation

Testing waived 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 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
Bacillus 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 LD50 for Bacillus pumilus strain BU F33 was greater than 5,000 mg/kg body weight or 

3.19 x 1012 cfu/kg body weight per day (1.6 x 1013

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 

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/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: Acceptable

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 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 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 Use –

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 systemic 

resistance in plants, 

thereby enhancing their 

defense against a 

spectrum of plant 

pathogens, including 

viruses, fungi, and 

bacteria.