Bacillus pumilus strain GHA 180 Page 1 of 27 Biopesticides Registration Action Document

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BIOPESTICIDES REGISTRATION ACTION DOCUMENT



### Bacillus pumilus strain GHA 180 Pesticide Chemical (PC) Code: 016485



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U.S. Environmental Protection Agency Office of Pesticide Programs

Bi opesticides and Pollution Prevention Division

February 10, 2012Bacillus pumilus strain GHA180 Page 2 of 27 Biopesticides Registration Action Document

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TABLE OF CONTENTS

I. EXECUTIVE SUMMARY

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

II. ACTIVE INGREDIE NT O VERVIEW

................................ ................................ ................ 5

III. REGULATORY BACKGROUN D

................................ ................................ ................. 5

A. Applications for Pesticide Registration

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

B. Food Tolerance Exemption

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

IV. RISK ASSESSMENT

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

A. Product Analysis Assessment (40 CFR § 158.2120)

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

B. Human Health Assessment (40 CFR § 158.2140)

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

C. Environmental Assessment (40 CFR § 158.2150)

................................ ................................ .............................. 12

V. ENVIRONMENTAL JU STICE

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

VI. RISK MANAGEMENT DECISION

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

VII. ACTIONS REQUIR ED BY T HE REGISTRANT

................................ ......................... 14

VIII. GLOSSARY OF A CRONYMS AND ABBREVIA TIONS

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

A. Studies Submitted to Support the Bacillus pumilus GHA180 Pesticide Product Registrations

.................. 16

B. Environmental Protection Agency Risk Assessment Memoranda

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

C. Other References

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

APPENDIX A. MICROBI AL PESTICIDES DATA R EQUIREMENTS

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

APPENDIX B. PESTICI DE PRODUCTS

................................ ................................ ............... 27Bacillus pumilus strain GHA180 Page 3 of 27 Biopesticides Registration Action Document

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BIOP ESTICIDES REGISTRATION ACTION DOCUMENT TEAM

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

Microbial Pesticides Branch

Science Reviews

Joel V. Gagliardi, Ph.D., Microbial Ecologist Product Analysis, Human Health

Joh n L. Kough, Ph.D. Product Analysis, Human Health

Gail Tomimatsu , Ph.D. Environmental Effects

Regulations

Sheryl K. Reilly, Ph.D. Chief, Microbial Pesticides Branch Alan Reynolds Team Leader, Microbial Pesticides Branch

Susanne Cerrelli Reg ulatory Action LeaderBacillus pumilus strain GHA180 Page 4 of 27 Biopesticides Registration Action Document

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I. EXECUTIVE SUMMARY

Bacillus pumilus strain GHA180 is a new microbial active ingredient discovered in an apple tree rhizosphere in Mexico. This active ingredient is a bacteri um that produces metabolites that suppress the fungal plan t pathogens Pythium ultimum, Fusarium oxysporum, Rhizoctonia solani

and Alternaria spp

. This bacterium colonizes plant roots, forming a biofilm.

Premier Horticulture Ltd. h as proposed to register t hree end

- use pesticide product s (EPs) :

**•** GHA 180 Biologica l Fungicide

, an aqueous suspension, to incorporate into peat moss or growing media or as a soil drench in greenhouses and enclosed nurseries to suppress plant pathogens that attack root systems

. ( Contain s 0.37 % Bacillus pumilus strain
GHA180, a minimum of 1 x 10 9 viable spores/ml)

**•** Pro

- Mix GHA180 and Pro

- Mix Tandem. B o th Pro

- Mix products are a series of growing media formulations that contain Bacillus pumilus strain GHA180 to suppress plant pathogens that attack root systems. (Both Pro

- Mix products contai n Bacillus pumilus strain GHA180 at 0.001%, with a minimum of 5 x 10 4 viable spores per a gram

of media

. )

P ro

- M ix GHA180 and Pro

- Mix Tandem are each formulated into eight different custom blended growing mixes: BX, HP, TA, FPX, MPX, PGX, VFT and BRK Blend s . There are 16 different blended growing mixes in total

.

Environmental Protection Agency (EPA) scientists have reviewed product analysis, toxicology, and nontarget organism data and information (40 Code of Federal Regulations (CFR) §§ 158.2120, 158.2140, and 158.2150, respectively) submitted to support the registration of th e three proposed Bacillus pumilus strain GHA 180 products

. Overall, these data and information are adequate for risk assessment purposes , fulfill current data requirements

, and allow fo r registration under Section 3(c)(5) of the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA)

.

Acceptable guideline studies to support the proposed use of these products fulfilled p roduct analysis data requirements for Bacillus pumilus strain GH A 180 , including product chemistry and composition, analysis of samples, and physical and chemical characteri stics

.

Adequate mamm alian toxicology data and information were submitted to support Bacillus pumilus strain GHA 180 pesticide products.

EPA has concluded that there is a reasonable certainty that no harm will result to the United States population, including infants and children, from aggregate exposure to residues of Bacillus pumilus strain GHA 180. No dietary risks are expected from use of Baci llus pumilus strain GHA

180 as an active ingredient in pesticide products.

The proposed use sites for Bacillus pumilus strain GHA180 are limited to greenhouse and

contained nurseries . I t was determined that the use sites for Bacillus pumilus strain GHA180 on the proposed labels would not result in significant nontarget species exposure and did not trigger the nontarget and environmental fate date requirements under 40 CFR § 158.2150

.Bacillus pumilus strain GHA180 Page 5 of 27 Biopesticides Registration Action Document

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On October 1, 2009, EPA announced a new policy to provide a more meaningful opportunity for the public to participate in major registration decisions before they occur. Under this policy, EPA provide s 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 EPA believes there may be significant public interest.

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

pe sticide products containing the new active ingredient, Bacillus pu milus strain GHA 180, are subject to a 30

- day comment period. In addition to containing a new active ingredient, the registration of each of these products would result in the first food use for Bacillus pumilus strain

GHA 180. The docket identification n umbers associated with these registration actions and accessed through either http://www.regulations.gov/ or http://www.epa.gov/pestici des/regulating/registration

- status.html

, are EPA

– HQ

– OPP

– 2010

– 0499 for Bacillus pumilus strain GHA180 products

. The following documents are available for

comment in EPA

– HQ

– OPP

– 2010

– 0499: 1) draft Bacillus pumilus strain GHA180 Biopesticides Registration Action Document (BRAD) ; (2) draft product label s for the end

- use product s
, GHA 180 Biological Fungicide (EPA File Symbol 74267

- E

, Pro

- Mix ‘ GHA180’ (EPA File Symbol

74267

- G ) , and Pro

- Mix Tandem (EPA File Symbol 74267

- U) . W hile a final decision on registrati on is contingent upon review and consideration of public comments, EPA presently

believes t hat, the risk assessment and informat ion submitted support registration of the GHA 180 Biological Fungicide, Pro

- Mix ‘ GHA180’, and Pro

- Mix Tandem EPs

. The basis for this preliminary decision can be found in the risk assessment for Bacillus pumilus strain GHA 180

, which is characterized throughout this BRAD.

II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Bacillus pumilus strain GHA 180

Culture Deposit: Inte rnational Depository Authority of Canada, under the reference

number IDAC No. 260707

- 01

OPP Chemical Code: 016485

Type of Pesticide: Microbial Pesticide

– Fungicide

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

us e sites, application rate s , methods of application, formulation type s , and target pests) regarding the registered pesticid e products containing this active ingredient.

III. REGULATORY BACKGROUNDBacillus pumilus strain GHA180 Page 6 of 27 Biopesticides Registration Action Document

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A. Applications for Pesticide Registration

On Feb ruary 14, 2010 , Premier Horticulture , (address: 1, avenue Premier, Rivie`re

- du

- Loup, Quebec, Canada G5R6C1 ) submitted applications to register three end

- use products, GHA 180 Biological Fungicide (EPA File Symbol 74267

– E), Pro

- Mix GHA 180 (EPA File Symbol 7 4267

– G) and Pro

- Mix Tandem (EPA File Symbol 74267

– U) under FIFRA section 3

. At that time

Premier Horticulture applied to register GHA 180 Biological Fungicide (EPA File Symbol

74267

– E) also as a manufacturing product. On November 24, 2010

, EPA announced r eceipt of

these application s to register pesticide product s containing a new active ingredient ( 75 Federal Register (FR) 71698 ) and opened a 3 0

- 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 application s and under Federal Food, Drug, and Cosmetic Act (FFDCA) section 408(d), Premier Horticul tur e submitted a petition to establish an exemption from the requirement of a tolerance for Bacillus pumilus GHA180 (Pesticide Petition (PP)
0F7698)

. In the Federal Register of September 30, 2010 ( 75 FR 60452 ), EPA announced that

Premier Horticulture proposed to establish an exemption from the requirement of a tolerance for

residues of the fungicide, Bacillus pumilus GHA180 in or on all agricultural commodities and water systems when app lied as described in the draft label and opened a 30

- day comment period. No comments were received following this publication.

IV. RISK ASSESSMENT

In the Federal Register of October 26, 2007, EPA issued a Final Rule on the data requirements to

support registration of microbial pesticides and updated the definition for microbial pesticides ( 72 FR 61002 )

. The rule became effective on December 26, 2007. The data and information

eval uated for this BRAD were considered in light of these requirements.

The classifications for each data submission are assigned by EPA science reviewers and i ndicat e 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 t o be scientifically acceptable (“ supplemental : upgradeable ”). If a study is rated as “supplemental: upgradeable ,” 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 “ suppl emental ” rating, the reviewer often will state that t he study is not required by 40 C FR P art 158. Both “ acceptable” and “supplemental ” studies may be used in the risk assessment process as appropriate. An “unacceptable ” rating indicates that new

data need to 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 EPA in support of a pesticide registration. The active ingredient or particul ar product is classified as Toxicity Category I, II, III, or IV, where Toxicity Category I indicates the highest toxicity and Toxicity Category IV indicates the lowest toxicity.Bacillus pumilus strain GHA180 Page 7 of 27 Biopesticides Registration Action Document

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A. Product Analysis Assessment ( 40 CFR § 158.2120 )

Bacillus pumilus strain GHA180 is a new microbial active ingredient discovered in an apple tree rhizosphere in Mexico. This b acteri um produces metabol i tes that suppress the fungal plant pathogens Pythium ultimum, Fusarium oxysporum, Rhizoctonia solani and Alternaria spp. and colonizes plant roots, forming a biofilm

. A ll product analysis data requirements for Bacillus pumilus st rain GHA180 were fulfilled to support current use sites . Refer to Tables 1 , 2, 3, 4 , 5,

and 6 in Appendix A for a brief summary of the data requirements, including both generic and product

- specific information.

B. Human Health Assessment ( 40 CFR § 158.2140 )

1. Toxicity

All toxicology data requirements for Bacillus pumilu s strain GHA180 were fulfilled

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

strain GHA180 pesticide products

. Furthermore, Tier II and Tier III studies were not required for Bacillus pumilus stra in GHA180 b ecause of the lack of acute toxicity/pathogenicity in the Tier I

studies.

For a compr ehensive 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 Table 7 in Appendix A

.

a. Acute Toxicity/Pathogenicity

– Tier I

Acute Oral Toxicity/Pathogenicity ( Office of Chemical Safety and Pollution Prevention (OCSPP)

Guideline 8 85.3050; and Acute Pulmonary Toxicity/Pathogenicity (OCSPP Guideline
885.3150; Master Record Identification Number (MRID No 48005019

. ) : W aiver rationale for a cute o ral and pulmonary toxicity/pathogenicity data requirements w ere submitted by Premier Horticu lture Ltd , b ased on the acute injection toxicity and pathogenicity

stud y and the oral, dermal and inhalation acute toxicity study results discussed below

. All of

those tests show ed no evidence of infectivity, pathogenicity

, toxicity , or other adverse effec ts.

EPA determined that in light of these results, adverse effects from exposure to Bacillus pumilus

GHA180 are not expected, and waived the ac ute oral toxicity/pathogenicity and acute pulmonary

t oxicity/pathogenicity data requirements for this active ingr edient.

Acute Injection Toxicity/Pathogenicity (OCSPP Guideline 885.3200; MRID No. 48005025 ) : Thirty young adult rats (15/ sex) were injected intravenously with Bacillus pumilus GHA180 TGAI (1.4×10 10

– 6.0×10 10 CFU/mL) at a dose of 6.8×10 7 CFU/animal . Two c ontrol groups (5/sex/treatment) were treated with inactivated Bacillus pumilus GHA180 TGAI

, and saline.The

animals were observed for up to 21 days. Bacterial cell counts were made on the blood, liver, kidney, spleen, lungs, mandibular lymph node, brain, an d cecum contents of the rats given live

bacteria on study d ays 0, 7, 14 and 21, and on d ay 21 for the control groups. All animals gained weight and a ppeared normal during the study, and n o visible ab nor malities were noted at necropsy. The test organism c ompletely cleared from the blood, kidneys, mandibular lymph nodes and cecum contents of the animals by day 14, and it showed a pattern of clearance but had not completely cleared from the liver and spleen by day 21. No Bacillus pumilus GHA180 was detected in the tissues/ organs of the animals treated with the inactivated test material or the untreated control group. Bacillus pumilus GHA180 was not toxic, infective or pathogenic to ratsBacillus pumilus strain GHA180 Page 8 of 27 Biopesticides Registration Action Document

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when dosed intravenously at 6.8 × 10 7 CFU/animal. This study was classifie d as Acceptable.

Hypersensitivity Incidents (OCSPP Guideline 885.3400 ) : No hypersensitivity incidents, including immediate

- type or delayed

- type reactions of humans and domestic animals during research, development, or testing of Bacillus pumilus GHA180

w ere reported by the applicant . Any future hypersensitivity incidents must be reported per

OCSPP Guideline 885.3400.

Cell Culture (OCSPP Guideline 885.3500) : This study was not required because Bacillus pumilus strain GHA180 is not a virus (refer to test note #4 of 40 CFR § 158.2140(d)).

Acute Oral Toxicity (OCSPP Guideline 870.1100 ; MRID No. 48005020 ) : Using the up and down method, 3 fasted young adult female rats were given a single oral gavage dose of Bacillus pumilus GHA180 TGAI ( 1.4×10 10

– 6.0×10 10 CFU/mL ) at a concentration of 5000 mg/kg body weight (bw), and observed for 14 days. All animals gained weight and appeared normal throughout the study. No visible abnormalities were found at necropsy. The

LD 50 > 5000 mg/kg bw

. This study was classified as Acceptable; EPA Toxicity Category IV.

Acute Dermal Toxicity (OCSPP Guideline 870.1200 ; MRID 48005021 ) : Ten young adult rats (5/sex) were exposed to 5050 mg/kg of Bacillus pumilus GHA180 TGAI (1.4×10 10

– 6.0×10 10 CFU/mL ) for 24 hours on a clipped area of skin ( ~10% of the body surface area ), and observed for 14 days. One female died on day 1 ; at necropsy, mottled lungs and a def ormed liver were noted but

not attributed to the test substance. All other animals gained weight and appeared normal during the study. No dermal irritation resulted from exposure to the test material, and observable abnormalities were not found at necropsy

. The LD 50 was > 5050 mg/kg bw. This study was classified as Acceptable; EPA Toxicity Category IV.

Acute Inhalation Toxicity (OCSPP Guideline 870.1300 ; MRID 48005022 ) : Groups of young adult rats (5/sex/group) were exposed nose

- only to Bacillus pumilus GHA180 TGAI (1.4×10 10

– 6.0×10 10 CFU/mL ) aerosol for 4 hours at a concentration of 2.18 mg/L, then observed for 14 days. All anim als gained weight and appeared normal throughout the study. No abnormalities were

observed at necropsy. The LC 50 > 2.18 mg/L. This study was classified as Acceptable; EPA Toxicity Category IV.

Acute Eye Irritation (OCSPP Guideline 870.2400; MRID 48005023 ) : Three New Zealand White rabbits (2 male, 1 female) were exposed to 0.1 mL of undiluted Bacillus pumilus GHA180 TGAI (1.4x10 10

– 6.0x10 10 CFU/mL ; instilled into conjunctival sac of the right eye ) and observed for eye irritation at 1, 24, 48, and 72 hours after exposure

. Corneal opacitiy was observed in 1 male at 1 hour post exposure, which cleared by 48 hours. No iritis or conjunctival irritation was noted in any animal throughout the study. The maximum average

score was 17.3 at 1 hour after exposure

. Bac illus pumilus GHA180 TGAI was mildly irritating to the eye

. This study was classified as Acceptable; EPA Toxicity Category I II.

Primary Dermal Irritation (OCSPP Guideline 870.2500 ; MRID 480050

- 24 ) : The skin of 3 rabbits (1 male , 2 female ) were exposed to 0.5 mL (under a patch) of Bacillus pumilus GHA180 TGAI (1.4×10 10

– 6.0×10 10 CFU/mL) for 4 hours on an approximately 2.5 × 2.5 cm area of the body surface. The animals were observed at 1, 24, 48, and 72 hours after patch removal. Very slight erythema was no ted on 1 animal at 1 hour after patch removal ; this clearedBacillus pumilus strain GHA180 Page 9 of 27 Biopesticides Registration Action Document

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by 24 hours. No other rabbit exhibited skin irritation during the study . The primary irritation

index was 0.1. This study was classified as Acceptable; EPA Toxicity Category IV.

b. Acute Toxico logy and Subchronic Toxicity/Pathogenicity

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

– Tier III

Tier II and Tier III studies were not required for Bacillus pumilus GHA 180 based on the lack of acute toxicity/pathogenicity in the Tier I studies.

c. Endocrine Disruptors

As required under FFDCA section 408(p), EPA has developed the Endocrine Disruptor Screening Program (EDSP) 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

des ignate.” The EDSP employs a two

- tiered approach to making the statutorily required determinations. Tier 1 consists of a battery of 11 screening assays to identify the potential of a chemical substance to interact with the estrogen, androgen, or thyroid (E, A, or T) hormonal systems. Chemicals that go through Tier 1 screening and are found to have the potential to interact with E, A, or T hormonal systems will proceed to the next stage of the EDSP wh ere EPA will determine which, if any, of the Tier 2 tests a re necessary based on the available data. Tier 2 testing is designed to identify any adverse endocrine

- related effects caused by the substance, and establish a quantitative relationship between the dose and the E, A, or T effect.

Between October 2009 a nd February 2010, EPA issued test orders/data call

- ins for the first group of 67 chemicals, which contains 58 pesticide active ingredients and 9 inert ingredients. This list of chemicals was selected based on the potential for human exposure through pathways such as food and water, residential activity, and certain post

- application agricultural scenarios. This list should not be construed as a list of known or likely endocrine disruptors.

Bacillus pumilus GHA 180 is not among the group of 58 pesticide active ingredients on the initial list to be screened under the EDSP. Under FFDCA section 408(p), EPA must screen all pesti cide chemicals. Accordingly, EPA anticipates issuing future EDSP orders/data call

- ins for all pesticide active ingredients.

For further in formation 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: http://www.epa.gov/endo/

.

2

. Federal Food, Drug, and Cosmetic Act (F FDCA ) Considerations

Section 408(c)(2)(A)(i) of FFDCA allows EPA to establish an exemption from the requirement for a tolerance (the legal limit for a pesticide chemical resi due in or on a food) only if EPA

determines that the exe mption 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 e xposures for which there is reliable information.” This includes exposure through drinking water and in residential settings but does not include occupational exposure. Pursuant to section 408(c)(2)(B)Bacillus pumilus strain GHA180 Page 10 of 27 Biopesticides Registration Action Document

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of FFDCA, in establishing or maintaining in effect an exemption from the requirement of a tolerance, EPA must take into account the factors set forth in section 408(b)(2 )(C) of FFDCA, which require EPA to give special consideration to exposure of infants and children to the pesticide chemical residue in estab lishing a tolerance 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, section 408(b)(2)(D) of FFDCA requires that EPA consider

“ available information concerning the cumulative effects of [a particular pesticide's] residues and other substances that have a common mechanism of toxicity.”

EPA performs a number of analyses to determine the risks from aggregate exposure t o pesticide

res idues. First, EPA determines the tox icity of pesticides. Second, 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 s ection 408(b )(2)(D) of FFDCA, EPA has reviewed the available scientific data and other relevant information, and considered its validity, completeness, and reliability and the relationship of thi s information to human risk. EPA has also considered available informatio n concerning the variability of the sensitivities of major identifiable subgroups of consumers, including infants and children.

Based on the acute toxicity/pathogenicity data and information discussed previously and presented in Table 7 in Appendix A , t he data required for a FQPA risk assessment for Bacillus pumilus strain GHA 180 have been fulfilled

. Hence, EPA determined that the exemption is

“safe”.

a. Aggregate Exposure

In examining aggregate exposure, section 408 of FFDCA directs EPA to consider available information concerning exposures from the pesticide residue in food and all other nonoccupational exposures, including drinking water from ground water or surface water and exposure through pesticide use in gardens, lawn s, or buildings (residential and other indoor uses).

Dietary Exposure and Risk Characterization : Bacillus spp. including Bacillus pumilus are commonly found in agricultural settings (i.e. soils) and are present on fresh produce of all kinds

with no known adverse effects. The Manual of Clinical Microbiology (9 th edition) mentions that dried food such as spices, milk powder and grains often contain large amounts of Bacillus

spores. Bacillus pumilus has been described as a natural, halotolerant component of fermented

fish sauce and in cocoa bean fermentations (Doyle et. al, 1997)

. Bacillus pumilus strain GHA180 is not known to produce any mammalian toxins

, and no foodborne disease outbreaks associated

with Bacillus pumilus strain GHA180 or foodborne toxin pro duction from Bacillus pumilus

strain GHA180 have been reported. Based on extensive testing of Bacillus pumilus strain

GHA180 TGAI , no toxicity, infectivity, pathogenicity or other adverse effects from dietary

exposure to Bacillus pumilus strain GHA180 are expected. Even if oral exposure should occur by ingesting harvested fruit vegetables, and/ or tobacco

, EPA concludes that there is a reasonable certainty that no harm will result from exposure to such residues because of the lack of toxicity, infectivity, and or pathogenicity in the previously described toxicological studies (see section IV(B)(1)(a) and Table 7 in Appendix A ).

Drinking Water Risk Characterization : Since Bacillus pumilus is naturally present in soils,Bacillus pumilus strain GHA180 Page 11 of 27 Biopesticides Registration Action Document

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exposure to Bacillus pumilus from surface and possibly groundwater can be expected. No adverse effects are expected or have been reported from exposure to Bacillus pumilus through drinking water

. According to the World Health Organization, Bacillus species are often d etected 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)

. Should this microbial pesticide be present in drinking wate r , no adverse effects are expected from exposure to Bacillus pumilus GHA180 through drinking water, based on the results of toxicity studies described in section (B)(1)(a) and Table 7 in Appendix A

.

Non

- occupational, Residentia l Risk Characterization : The use sites for these products include greenhouse and contained nurseries . As stated previously, Bacillus pumilus is naturally present in many habitats . B ased on the studies and other information submitted to satisfy data requir ements for registration of the end

- use pesticide products containing the active ingredient Bacillus pumilus strain GHA180, no toxicity, infectivity, pathogenicity or other adverse effects from

non

- occupational exposure are expected (see section IV(B)(1)(a ), above, and Table 7 in Appendix A ).

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 tole rance exemption

, EPA consider “available information concerning the cumulative effects of [a particular pesticide’s] residues and other substances that have a common mechanism of toxicity.”

EPA has not found Bacillus pumilus strain GHA180 to share a common mechanism of toxicity with any other substance, and Bacillus pumilus strain GHA180 do es not appear to produce a toxic metabolite as its mode of action against target pest s

. Therefore, f or the purposes of this tolerance action, EPA has assumed that Bacillu s pumilus strain GHA180 do es not have a common mechanism of toxicity with other substances. For information on EPA ’s efforts to determine which chemicals have a common mechanism of toxicity and to evaluate the

cumulative ef fects o f such chemicals, see EPA ’ s website at http://www.epa.gov/pesticides/cumulative

.

c. Determination of Safety for the U.S. Population, Infants and Children

FFDCA sectio n 408(b)(2)(C) provides that EPA shall assess the availa ble 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 mech anism of toxicity. In addition, FFDCA sectio n 408(b)(2)(C) provides that 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 complet en ess of the database on toxicity and exposure unless 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 FQPA Safety Factor. In applying this provision, EPA either retains the default value of 10X or uses a different additional safety factor when reliable data available to EPA support the choice of a different factor.

Based on the acute toxicity and pathogenicity data summarized in section IV(B)(1)(a) and Tabl e
7 in Appendix A

, EPA concludes that there is a reasonable certainty that no harm will result toBacillus pumilus strain GHA180 Page 12 of 27 Biopesticides Registration Action Document

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the U.S. population, including infants and children, from aggregate exposure to the residues of Bacillus pumilus strain GHA180. Thi s includes all anticipated dietary exposures and all other exposures for which there is reliable information. EPA has arrived at this conclusion because the

data and information available on Bacillus pumilus strain GHA180 do es not demonstrate toxic, pathog enic, and/ or infective potential to mammals. Because there are no threshold effects of concern, the provision requiring an additional margin of safety does not apply.

3. Occupational Exposure and Risk Characterization

Handler exposure to Bacillus pumil us strain GHA180 is not expected to pose any undue risk.

A ppropriate personal protective equipment and precautionary statements are required on pesticide product labels to mitigate any potential risks to pesticide handlers due to prolonged or

numerous exposures. Handlers applying Bacillus pumilus strain GHA180 liquid end

- use products in agricultural settings must wear a long

- sleeved shirt, long pants, socks, shoes, waterproof gloves, and a dust/mist filtering respirator meeting NIOSH standards of at lea st N

- 95,

R

- 95, or P

- 95. Additional PPE, other than the standard described above, may be required on a product

- specific basis.

4. Human Health Risk Characterization

EPA considered human exposure to Bacillus pumilus strain in light of the standard for regi stration in FIFRA and the relevant safety factors in FFDCA. A determination has been made

that no unreasonab le adverse effects to the U.S. population in general, and to infants and children in particu lar, will result when Bacillus pumilus strain GHA180 pes ticide products are used in

accordance with EPA

- approved labeling.

C. Environmental Assessment ( 40 CFR § 158.2150 )

The Agency has performed a n environmental risk assessment for the proposed uses of Bacillus pumilus strain GHA180 . T he proposed use sites for Bacillus pumilus strain GHA180 were

limited to greenhouse and contained nurseries

. EPA determi ned that these use sites for Bacillus pumilus strain GHA180 would not result in significant nontarget species exposure and d id not trigger the nontarget and environmental fate date requirements under 40 CFR § 158.2150

. Further testing of nontarget organisms at higher tier levels (i.e., Tiers II, III, and IV) is not required for the current uses and app lication methods.

For a comprehensive summary of the generic data requirements described in sections IV(C)(1),

refer to Table 8 in Appendix A

.

1

. Threatened and Endangered Species Assessment

A “No Effect” determination is al so made for direct and indirect effects to listed species and their designated critical habitats from proposed uses of Bacillus pumilus strain GHA180 as labeled.

The exposure of nontarget organisms to Bacillus pumilus strain GHA180 is not expected to

incre ase the amount of Bacillus pumilus above levels that are found naturally in the environment because the use of products containing this active ingredient are limited to green house and

enclosed nurseries. Further testing of nontarget organisms at higher tier levels is not required.

B ased on the data, literature citations, and data waiver rationale provided by the applicant

, BPPDBacillus pumilus strain GHA180 Page 13 of 27 Biopesticides Registration Action Document

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has determined that adverse effects to nontarget organisms are not anticipated from the proposed

uses of Bacillus pumilus strai n GHA180

.

V. ENVIRONMENTAL JUSTICE

EPA seeks to achieve environmental justice

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

— with respect to the development, implementation, and enforce ment 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

industria l, 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; (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. EPA has this goal for all communities and persons

across the United States.

To help address potential environmental justice issues, EPA seeks info rmation 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 GHA180 , compared to the general population. Please comment if y ou 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 EPA’s web site at http://www.epa.gov/compliance/environmentaljustice/index.html

.

V I

. RISK MANAGEMENT DECISION

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

(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 e ffects 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 Bacillus pumilus str ain GHA180 pesticide products have well

- known properties. EPA has no knowledge that would contradict the claims made on the GHA 180 Biological Fungicide

, Pro

- Mix ‘ GHA180’, and Pro

- Mix Tandem EP label s . These products are not expected to cause unreasonable adverse effects on the environment when used according to label instructions. Criterion 2 is satisfied by the current product label s, as well as the data and

information presented in this document. It is believed that the Bacillus pumilus strain GHA180 1

w ill not cause any unreasonable adverse effects on the environment, and the GHA 180Bacillus pumilus strain GHA180 Page 14 of 27 Biopesticides Registration Action Document

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Biological Fungicide

, Pro

- Mix ‘ GHA180’, and Pro

- Mix Tandem EP s (end

- use product s ) , in particular, are likely to protect against fungal pests as claimed, thereby satisfying c riterion 3.

Criterion 4 is satisfied in that the Bacillus pumilus strain GHA180 pesticide products are not expected to cause unreasonable adverse effects when used according to label instructions. Therefore, the GHA 180 Biological Fungicide, Pro

- Mix ‘ GHA18 0’, and Pro

- Mix Tandem (all containing Bacillus pumilus strain GHA180 as a new active ingredient ) are eligible for registration under FIFRA section 3(c)(5) for labeled uses. If the applicant proposes uses that are

more extensive in the future (e.g., outdo or or aquatic applications )

, EPA will likely require that

additional data be submitted in conjunction with a registration amendment request

.

V I I

. ACTIONS REQUIRED BY THE REGISTRANT

A. Final Printed Labeling

Before releasing pesticide products containi ng Bacillus pumilus strain GHA180 for shipment, the registrant is required to provide appropriate final printed labeling to EPA

.

B . Reporting of Adverse Effects and Hypersensitivity Incidents

Notwithstanding the information stated in the previous sectio ns, it should be clearly understood that certain, specific data are required to be reported to 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 al l incidents of adverse effects to the environment must be submitted to 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 EPA unde r the provisions of 40 CFR § 158.2140(d).Bacillus pumilus strain GHA180 Page 15 of 27 Biopesticides Registration Action Document

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VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

ASAE American Society of Agricultural Engineers

BPPD Biopesticides and Pollution Prevention Division

BRAD Biopesticides Registration Action Document

bw body we ight of animal

CFR Code of Federal Regulations

c fu colony

- forming unit (s) cfu/kg colony

- forming units per kilogram

cfu/mL colony

- forming unit s per milliliter

cP centipoise

DSMZ Deutsche Sammlung von Mikroorganismen und Zellkulturen (the German Strain

Coll ection for Microorganisms)

EDSP Endocrine Disruptor Screening Program

EP end

- use product

EPA Environmental Protection Agency (the “Agency”)

FFDCA Federal Food, Drug, and Cosmetic Act

FIFRA Federal Insecticide, Fungicide, and Rodenticide Act FQPA Food Qual ity Protection Act

FR Federal Register g/ mL gram per milliliter

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 (o ral, 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 milligram s per liter

MP manufacturing

- use product

MRID No. Master Record Identification Number

NIOSH Nation al 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 protecti ve equipment

ppm parts per million

TGAI technical grade of the active ingredient

ULV ultra low volumeBacillus pumilus strain GHA180 Page 16 of 27 Biopesticides Registration Action Document

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I X. BIBLIOGRAPHY

A. Studies Submitted to Support the Bacillus pumilus GHA180 Pesticide Product Registrations

MRID Citation Reference

48005000 Premier Horticulture (2010) Submission of Product Chemistry and Toxicity Data in Support of the Application for Registration of GHA 180, Pro

- Mix GHA 180,

and Pro

- Mix Tandem and the Petition for Tolerance of Bacillus pumi lus GHA

180. Transmittal of 25 Studies.

48005001 Libman, G. (2009) Product Chemistry for ( Bacillus pumilus GHA 180). Unpublished study prepared by Premier Tech, Ltd. 33 p.

48005002 Libman, G. (2009) Product Chemistry for PRO

- MIX 'BX' Tandem. Project

Nu mber: PHL/LA/WI/137E3/USA. Unpublished study prepared by Premier Tech, Ltd. 77 p.

48005003 Libman, G. (2010) Product Chemistry for Pro

- Mix GHA 180 'BX' Blend. Unpublished study prepared by Premier Tech, Ltd. 89 p.

48005004 Libman, G. (2010) Product Chemi stry for Pro

- Mix GHA 180 'HP' Blend. Unpublished study prepared by Premier Tech, Ltd. 80 p.

48005005 Libman, G. (2010) Product Chemistry for Pro

- Mix GHA 180 'TA' Blend. Unpublished study prepared by Premier Tech, Ltd. 92 p.

48005006 Libman, G. (2010) Pro duct Chemistry for Pro

- Mix GHA 180 'FPX' Blend. Unpublished study prepared by Premier Tech, Ltd. 90 p.

48005007 Libman, G. (2010) Product Chemistry for Pro

- Mix GHA 180 'MPX' Blend. Unpublished study prepared by Premier Tech, Ltd. 80 p.

48005008 Libman, G . (2010) Product Chemistry for Pro

- Mix GHA 180 'PGX' Blend. Unpublished study prepared by Premier Tech, Ltd. 92 p.

48005009 Libman, G. (2010) Product Chemistry for Pro

- Mix GHA 180 'VFT' Blend. Unpublished study prepared by Premier Tech, Ltd. 92 p.

480050 10 Libman, G. (2010) Product Chemistry for Pro

- Mix GHA 180 'BRK' Blend. Unpublished study prepared by GNL Consultation Services and Premier Tech, Ltd. 80 p.

48005011 Libman, G. (2010) Product Chemistry for Pro

- Mix Tandem 'BX' Blend. Unpublished study prep ared by Premier Tech, Ltd. 126 p.

48005012 Libman, G. (2010) Product Chemistry for Pro

- Mix Tandem 'HP' Blend. Unpublished study prepared by Premier Tech, Ltd. 118 p.

48005013 Libman, G. (2010) Product Chemistry for Pro

- Mix Tandem 'TA' Blend. Unpublished study prepared by Premier Tech, Ltd. 94 p.

48005014 Libman, G. (2010) Product Chemistry for Pro

- Mix Tandem 'FPX' Blend. Unpublished study prepared by Premier Tech, Ltd. 92 p.

48005015 Libman, G. (2010) Product Chemistry for Pro

- Mix Tandem 'MPX' Blend.Bacillus pumilus strain GHA180 Page 17 of 27 Biopesticides Registration Action Document

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Un published study prepared by Premier Tech, Ltd. 82 p.

48005016 Libman, G. (2010) Product Chemistry for Pro

- Mix Tandem 'PGX' Blend. Unpublished study prepared by Premier Tech, Ltd. 94 p.

48005017 Libman, G. (2010) Product Chemistry for Pro

- Mix Tandem 'VFT' Blend. Unpublished study prepared by Premier Tech, Ltd. 94 p.

48005018 Libman, G. (2010) Product Chemistry for Pro

- Mix Tandem 'BRK' Blend. Unpublished study prepared by Premier Tech, Ltd. 77 p.

48005019 Libman, G. (2010) Waiver Request for Microbial Pes ticide Data Requirements: Bacillus pumilus GHA 180 and Formulated Products. Unpublished study prepared by Premier Horticulture. 35 p.

48005020 Kuhn, J. (2009) Acute Oral Toxicity Study (UDP) in Rats: Bacillus Pumilus

GHA

- 180 TGAI: Amended Final Report. Pr oject Number: 11547/08. Unpublished study prepared by Stillmeadow, Inc. 12 p.

48005021 Kuhn, J. (2010) Acute Dermal Toxicity Study in Rats: Bacillus pumilus GHA

- 180 TGAI: Amended Final Report. Project Number: 11548/08. Unpublished study prepared by Stillm eadow, Inc. 15 p.

48005022 Carter, L. (2009) Acute Inhalation Toxicity Study in Rats: Bacillus pumilus GHA

180 TGAI: Amended Final Report. Project Number: 11549/08. Unpublished study prepared by Stillmeadow, Inc. 19 p.

48005023 Kuhn, J. (2009) Acute Eye Irritation Study in Rabbits: Bacillus pumilus GHA

- 180 TGAI: Amended Final Report. Project Number: 11550/08. Unpublished study prepared by Stillmeadow, Inc. 18 p.

48005024 Kuhn, J. (2009) Acute Dermal Irritation Study in Rabbits: Bacillus pumilus GHA

180 T GAI: Amended Final Report. Project Number: 11551/08. Unpublished study prepared by Stillmeadow, Inc. 13 p.

48005025 Monds, K. (2009) Acute Intravenous Toxicity/Pathogenicity Study in Rats with a Microbial Pest Control Agent (MPCA): Bacillus pumilus GHA

- 18 0 TGAI: Amended Final Report. Project Number: 11552/08. Unpublished study prepared by Stillmeadow, Inc. 21 p.

48391500 Premier Horticulture, Inc. (2011) Submission of Product Chemistry Data in Support of the Applications for Registration of GHA 180 Biolog ical Fungicide,

Pro

- Mix GHA 180 and Pro

- Mix Tandem. Transmittal of 1 Study.

48391501 Libman, G. (2011) Addendum to Product Process Bacillus pumilus GHA 180 and Formulated Products: Addendum to MRID 48005001. Unpublished study prepared by: GNL Consultation Services and Premier Horticulture, Inc. 12 p.

48537300 Premier Horticulture, Inc. (2011) Submission of Product Chemistry Data in Support of the Application for Registration of Pro

- Mix GHA 180, GHA 180 Biological Fungicide and Pro

- Mix Tandem. Transmittal of 2 Studies.

48537302 Libman, G. (2011) Clarification about Storage Stability Data of Promix GHA 180 and Promix Tandem. Unpublished study prepared by Premier Horticulture, Ltd. 8 p.Bacillus pumilus strain GHA180 Page 18 of 27 Biopesticides Registration Action Document

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. 48537303 Libman, G. (2011) Microbiological Analyses: Certificates of A nalysis for GHA

- 180. Project Number: STIS/10109. Unpublished study prepared by Premier Horticulture, Ltd. 8 p.

48559501 Libman, G. (2011) 16S rRNA Gene Sequence and DNA Fingerprinting of the GHA 180 Bacterial Strain. Project Number: 1104081623E. Unpublis hed study prepared by GNL Consultation Services and Premier Horticulture, Inc. 32p.

48559502 Libman, G. (2011) Product Chemistry for (Bacillus pumilus GHA180). Project Number: PHL/LA/WI/128E2. Unpublished study prepared by GNL Consultation Services. 31p.

B. Environmental Protection Agency Risk Assessment Memoranda

U.S. EPA. 2011a

. Bacillus pumilus strain GHA180. Memorandum from Joel V. Gagliardi, Ph.D., and J. L. Kough, Ph.D., to S. Cerrelli dated March 22, 2011

U.S. EPA. 2011b. Bacillus pumilu s strain GHA180. Review of deficiency responses. Memorandum from Joel V. Gagliardi, Ph.D., and J. L. Kough, Ph.D., to S. Cerrelli dated October
20, 2011.

C. Other References

Aunpad, R. and K. Na

- Bangchang . 2007. Pumilicin 4, a novel bacteriocin with anti

- MRSA and

anti

- VRE activity produced by newly isolated bacteria Bacillus pumilus strain WAPB4. Curr.

Microbiol. 55(4):308

- 13.

Babana, A.H., H. Bathily, F. Samaké, K. Maïga, D. Traoré and A. Dicko. 2011. Microbiological

Control of Bacterial Soft Rot C aused by Bacillus pumilus Od23 on Potato. British Microbiology Research Journal 1(3): 41

- 48.

Banerjee C ., C.I. Bustamante , R. Wharton , E. Talley and J.C. Wade . 1988. us Bacillus infections in patients with cancer. Arch. Intern. Med. 148(8):1769

- 74.

Bathily, H., A.H. Babana and F. Samaké. 2010. Bacillus pumilus, a new pathogen on potato tubers in storage in Mali. African Journal of Microbiology Research 4(20):2067

- 2071.

Bentur H.N

., A.M. Dalzell and F.A. Riordan . 2007. Central venous catheter infection with Bacillus pumilus in an immunocompetent child: a case report. Ann. Clin. Microbiol. Antimicrob. 6(1):12.

Bottone, E.J. and R.W. Peluso. 2003. Production by Bacillus pumilus (MSH) of an antifungal compound that is active against Mucoraceae and Aspergillus spe cies: preliminary report. J. Med.

Microbiol. 52: 69

- 74.

Doyle, M.P., L.R. Beuchat and T.J. Montville. 1997. Food Microbiology: Fundamentals and Frontiers. American Society for microbiology, Washington, D.C.Bacillus pumilus strain GHA180 Page 19 of 27 Biopesticides Registration Action Document

- DRAFT

Farhat H ., E. Chachaty

, S. Antoun

, G. Nitenberg and J.R. Zahar . 2008. Two cases of Bacillus infection and immunodepression. Med. Mal. Infect. 38(11):612

- 4.

Font, M.I., D.D.M. Bassimba, M.C. Cebrián, L.M. Molina and C. Jordá. 2009. First report

of Bacillus pumilus on Phaseolus vulgaris in Spain. N ew Disease Reports 19:54.

From, C

., V. Hormazabal and P.E. Granum . 2007. Food poisoning associated with pumilacidin

producing Bacillus pumilus in rice. I nt. J. Food Microbiol. 115(3):319

- 24.

From C ., R. Pukall , P. Schumann , V. Hormazábal and P.E. Granum . 2005. Toxin

- producing ability

among Bacillus spp. outside the Bacillus cereus group. Appl. Environ. Microbiol. 71(3):1178

- 83.

Galal, A.A., A.A. El

- Bana and J. Janse. 2006. Bacillus pumilus, A New Pathogen on Mango Plants. Egyptian Journal of Phytopathology . 34(1):17

- 29.

Kramer, J.H. and R.J. Gilbert. 1989. Bacillus cereus and other Bacillus species, p. 21

- 70. In M.P. Doyle (ed.), Foodborne Pathogens. Marcel Dekker, Inc., New York.

Meyer, A. and O. Gottheil.1901. In: Gottheil, O. Botanische beschreibung ei niger bodenbakterien. Zentralbl. Bakteriol. Parasitenkd. Infektionskr. Hyg. Abt. II 7:680

- 691.

Murray, P.R, et al., Manual of Clinical Microbiology. Washington

, D. C. : ASM Press; 9th edition,

2007.

Naruse N ., O. Tenmyo , S. Kobaru , H. Kamei , T. Miyaki , M. Konishi and T. Oki . 1990. Pumilacidin, a complex of new antiviral antibiotics. Production, isolation, chemical properties, structure and biological activity. J. Antibiot. (Tokyo) 43(3):267

- 80.

Ozkocaman V ., T. Ozcelik , R. Ali , F. Ozkalemkas , A. Ozkan , C. Ozakin , H. Akalin , A. Ursavas , F.

Coskun , B. Ener and A. Tunali . 2007. Bacillus spp. among hospitalized patients with haematological malignancies: cl inical features, epidemics and outcomes. Int. J. Food Microbiol. 115(3):319

- 24.

Saleh, O.I., P.

- I. Huang and J.

- S. Huang. 19 97. Bacillus pumilus, the cause of bacterial blotch of immature balady peach in Egypt. Journal of Phytopathology 145:44

Skerman, V.B.D., V. McGowan and P.H.A. Sneath (eds.). 1980. Approved lists of bacterial names. Int. J. Syst. Bacteriol. 30:225

- 420.

Tena D

., J.A. Martinez

- Torres

, M.T. Perez

- Pomata

, J.A. Sáez

- Nieto

, V. Rubio and J. Bisquert

. 2007. Cutaneous infection due to Bacillus pumilus : report of 3 cases. Clin. Infect. Dis. 44(4):40

- 2.

World Health Organization, Guidelines for Drinking

- water Quality.(2011) Fourth Edition

.Bacillus pumilus strain GHA180 Page 20 of 27 Biopesticides Registration Action Document

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A PPENDIX A. MICROBIAL PESTICIDES DATA REQUIREM ENTS (40 CFR PART 158

– SUBPART V )

TABLE 1. Product Analysis Data Requirements for the Technical Grade of the Active Ingredient (TGAI) / GHA 180 (40 CFR § 158.2 120 )

OCSPP

G uideline

N umber Data Requirement Results MRID No. TGAI GHA 180 ( E P )

885.1 100 Product Identity Submitted data fulfill the requirement for product identity. Submitted data fulfill the requirement for product identity. 48005001

48559501

885.1200 Manufacturing Process Submitted data fulfill the requirement for manufacturing

process. 48005001

48391501

Not a pplicable Deposition of a Sample in a Nationally Recognized Culture

Collection Bacillus pumilus strain GHA180 is on deposit at the International Depository Authority of Canada, IDAC

No. 260707

- 01 Not applicable 48005001

8 85.1300 Discussion of Formation

of Unintentional Ingredients Submitted data fulfill the requirement for discussion of formation of unintentional ingredients 48005001

885.1400 Analysis of Samples Submitted data fulfill the requirement for analysis of sampl es 48005001

48537303

885.1500 Certification of Limits Not applicable Limits listed on the

confidential statement of

formula are adequate/acceptable CSFs dated July 12,2011

Additional Studies

830 .1800 Enforcement Analytical

Method Not applicable Submitt ed data fulfill the requirement for an enforcement analytical

method 48005001Bacillus pumilus strain GHA180 Page 21 of 27 Biopesticides Registration Action Document

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TABLE 2 . Physical and Chemical Characteristics for the Technical Grade of the Active Ingredient ( (TGAI )/ GHA 180 Biological Fungicide (EP) (40 CFR § 158.2120)

OCSPP

G uidelin e N umber Data Requirement Results MRID No.

TGAI GHA 180 Biological Fungicide

( E P )

830.6302 Color Amber 48005001

48559502

830.6303 Physical State Liquid 48005001

48559502

830.6304 Odor Musty 48005001

48559502

830.6313 Stability to Normal and

Elev ated Temperatures, Metals,

and Metal Ions Vegetative cells expected to denature at >50 o C. 48005001

48559502

830.6317 Storage Stability Stable for at least one year; storage conditions not defined. 48005001

830.6319 Miscibility Not required per OCSPP 830 .1000(e)(2), the product is not an emulsifiable liquid

. 830.6320 Corrosion

Characteristics No corrosion observed during the storage stability testing 48005001

830.7000 pH 6

- 7. 48005001

48559502

830.7100 Viscosity Not applicable 2

- 5 mPa.s at 20 °C 48005001

830.7300 Density/Relative Density/Bulk Density

(Specific Gravity) 62.30 lbs/ft. 48005001

48559502Bacillus pumilus strain GHA180 Page 22 of 27 Biopesticides Registration Action Document

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TABLE 3

. Product Analysis Data Requirements for the End

- Use Product ( E P), Pro

- Mix GHA 180 (40 CFR § 158.2120)

OCSPP Guideline

Number Data Re quirement Results MRID No.

885.1100 Product Identity Submitt ed data fulfill the requirement for product identity

. 48005003

- 48005010

885.1200 Manufacturing Process Submitted data fulfill the requirement for manufacturing process. 48005003

- 48005010

4839150 1 Not applicable Deposition of a Sample in a Nationally Recognized

Culture Collection Not applicable Not applicable

885.1300 Discussion of Formation of Unintentional Ingredients Submitted data fulfill the requirement for

discussion of formation of uninte ntional ingredients for the current use sites.

. 48005003

- 48005010

885.1400 Analysis of Samples Submitted data fulfill the requirement for analysis of samples 48005003

- 48005010

885.1500 Certification of Limits Limits listed on the confidential stateme nt of formula are adequate/acceptable 48005003

- 48005010

CSFs dated

1/7/11

Additional Studies

830.1800 Enforcement Analytical

Method An analytical method is not required for enforcement purposes since the Agency is establishing an exemption from the requi rement of a tolerance without any numerical limitation. 48005003

- 48005010Bacillus pumilus strain GHA180 Page 23 of 27 Biopesticides Registration Action Document

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TABLE 4 . Product Analysis Data Requirements for the End

- Use Product (EP), Pro

- Mix Tandem (40 CFR § 158.2120)

OCSPP Guideline

Number Data Requirement Results MRID No.

885.110 0 Product Identity Submitted data fulfill the requirement for product identity. 48005011

- 48005018

885.1200 Manufacturing Process Submitted data fulfill the requirement for manufacturing process. 48005011

- 48005018

48391501 Not applicable Deposition of a Sample in a Nationally Recognized

Culture Collection Not applicable Not applicable

885.1300 Discussion of Formation of Unintentional Ingredients Submitted data fulfill the requirement for

discussion of formation of unintentional ingredients for the curren t use sites. 48005011

- 48005018

885.1400 Analysis of Samples Storage stability was shown up to two years in peat growing mixes stored outdoors with temperature variations from

- 30 to +30 o C 48005011

- 48005018

885.1500 Certification of Limits Limits listed on the confidential statement of formula are adequate/acceptable 48005011

- 48005018

CSFs dated

1/7/11

830.1800 Enforcement Analytical

Method An analytical method is not required for enforcement purposes since the Agency is establishing an exemption from the requirement of a tolerance without any

numerical limitation.

TABLE 5 . Physical and Chemical Characteristics for the End

- Use Product (EP), Pro

- Mix Tandem (40 CFR § 158.2120)

OCSPP Guideline

Number Data Requirement Results MRID No.

83 0.6317 Storage Stability Storage stability was shown up to two years in peat growing mixes stored outdoors with temperature variations

from

- 30 to +30 o C. 48005011

- 48005018

48537302

830.6319 Miscibility Not applicable, the product is not an

emulsifiable l iquid.

830.6320 Corrosion Characteristics Not Required

. The peat based soil mixes

were not considered corrosive material

830.7100 Viscosity Not applicable, the product is a solid

830.7300* Density/Relative Density/Bulk Density (Specific Gravity) 8

- 10 lbs/ft 3 CSFs dated

1/7/11Bacillus pumilus strain GHA180 Page 24 of 27 Biopesticides Registration Action Document

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TABLE 6 . Physical and Chemical Characteristics for the End

- Use Product (EP), Pro

- Mix GHA 180 (40 CFR § 158.2120)

OCSPP Guideline Number Data Requirement Results MRID No.

830.6317 Storage Stability Storage stability w as shown up to two years in peat growing mixes stored outdoors with temperature variations

from

- 30 to +30 o C. 48005003

- 48005010

48537302

830.6319 Miscibility Not applicable, the product is not an emulsifiable liquid.

830.6320 Corrosion Characteristics Not Required

. The peat based soil mixes

were not considered corrosive material

830.7100 Viscosity Not applicable, the product is a solid

830.7300* Density/Relative Density/Bulk Density (Specific Gravity) 8

- 10 lbs/ft 3 CSFs dated

1/7/11Bacillus pumilus strain GHA180 Page 25 of 27 Biopesticides Registration Action Document

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TA BLE 7 . Toxicology Data Requirements for the Technical Grade of the Active Ingredient (TGAI)/ and for the

End

- Use Products (EP), GHA 180 Biological Fungicide

, Pro

- Mix GHA 180 Pro

- Mix Tandem (40 CFR § 158.2140)

OCSPP

Guideline

Number Data Requirement T GAI GHA 180 Biological Fungicide (EP) Pro

- Mix

GHA 180 (EP) Pro

- Mix

GHA 180

Tandem (EP) MRID

No.

885.3050 Acute Oral Toxicity/ Pathogenicity No evidence of infectivity, pathogenicity or toxicity were noted in the Acute Injection study or in any of the

sub mitted acute studies listed below. Exposures through the pulmonary or oral routes are not expected to produce adverse effects based on existing test and product

characterization information. Not applicable Not applicable Not applicable 48005019

885.3150 A cute Pulmonary Toxicity/ Pathogenicity Not applicable Not applicable Not applicable

885.3200 Acute Injection Toxicity/ Pathogenicity 15 male and 15 female rats were injected with TGAI (1.4×10 10

– 6.0×10 10 CFU/mL) at a dose of 6.8×10 7

CFU/animal and ob served up to 21 days. All animals survived, gained weight, and appeared normal during the study. No observable ab normalities were noted in any animal at necropsy. The test organism completely cl eared from the blood, kidneys, mandibular lymph nodes and

ce cum contents of the animals by day 14, and sh owed a pattern of clearance but had not completely cleared from the liver and spleen by day 21. No test organism was

detected in the tissues/ organs of the animals treated with the inactivated test material or the untreated control group.

Classification: Acceptable

– not toxic, infective or pathogenic to rats when dosed by IV at 6.8 × 10 7 CFU/animal. Not applicable Not applicable Not applicable 48005025

885.3500 Cell Culture Not applicable Not applicable Not applicable Not applicable

870.1100 Acute Oral Toxicity Oral LD 50 was >5000 mg/kg bw.

Classification: Acceptable

TOXICITY CATEGORY IV Data on TGAI supports these EPs 48005020

870.1200 Acute Dermal Toxicity The LD 50 was >5050 mg/kg bw.

Classification: Acceptable

TOXICITY CATEGORY IV Data on

TGAI su pports these EPs 48005021

870.1300 Acute

Inhalation Toxicity The LC

50 Combined was > 2.18mg/L.

Classification: Acceptable

TOXICITY CATEGORY IV Data on TGAI supports these EPs 48005022

870.2400 Acute Eye

Irritation Two male and one female rabbits were tr eated with 0.1 mL

test material No iritis or positive conjunctival irritation was noted on any rabbit throughout the study. The maximum average score was 17.3 at one hour after test material

instillation

. Bacillus pumilus GHA180 TGAI was mildly irritating to the eye

. Classification: Acceptable

TOXICITY CATEGORY III Data on TGAI supports these EPs 48005023

870.2500 Primary

Dermal

Irritation One male and two female rabbits were dermally exposed to 0 Bacillus pumilus GHA180 TGAI (1.4×10 10

– 6.0×10 10 CFU/mL) Very slight erythema was noted on one animal one hour after patch removal with clearance by 24 hours. No dermal irritation was noted on any other rabbit during the study. The primary irritation

index was 0.1. CLASSIFICATION: Acceptable

TOXICIT Y CATEGORY IV Data on TGAI supports these EPs 48005024Bacillus pumilus strain GHA 180 Page 26 of 27 Biopesticides Registration Action Document

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TABLE 8

. Nontarget Organism Toxicity and Environmental Fate Data Requirements for the Technical Grade of the Active Ingredient (TGAI), Bacillus pumilus train GHA180

. (40 CFR § 158.2150)

OCSPP Guideline

Number Data Requirement Results MRID

No.

Tier I

885.4050 Avian Oral Toxicity Not required per 40 CFR § 158.2150

. T he current label use patterns for greenhouse and nursery use would not result in significant nontarget species

exposure. 4800501 9

885.4100 Avian Inhalation Toxicity/Pathogenicity

885.4150 Wild Mammal Toxicity/Pathogenicity

885.4200 Freshwater Fish Toxicity/Pathogenicity

885.4240 Freshwater InvertebrateToxicity/Pathogenicity

885.4280 Estuarine/Marine Fish and

Invertebra te 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 GHA180 based on the current uses and application methods.Bacillus pumilus strain GHA 180 Page 27 of 27 Biopesticides Registration Action Document

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APPENDIX B. PESTICIDE PRODUCTS

EPA

File Symb

ol Registration

Name Growing

Medium

Blends Percentage

Active Ingredient Formulation Type Use Site(s) Method(s)

of Application Application

Rate Target Pest

74267

-

E GHA 180 Biological Fungicide Not applicable 0.37% E nd

- use Product Greenhouses

And Enclosed

Nurseries Growing

media of

Ornamentals and vegetable

transplants Incorporatio

n Into Peat

Moss Or Growing

Media Soil

Drench As directed

on label Pythium

, Fusarium

and
Rhizoctonia

74267

-

G Pro

- Mix GHA

180 BX, HP, TA,
FPX, MPX, PGX, VFT

or BRK 0.001% End

- use Product Horticultural

and agricultural

plants

Tobacco germinating

seeds Plant in Growing

Medium Not applicable,

pre

- mixed potting mix Pythium,

Fusarium
Rhizoctonia

Alternaria Aspergillus

74267

-

U Pro

- Mix

Tandem B X, HP, TA,
FPX, MPX, PGX, VFT

or BRK 0.001% End

- use Product Horticultural

and agricultural

plants

Tobacco germinating

seeds Plant in Growing

Medium Not applicable,

pre

- mixed potting mix Pythium,

Fusarium
Rhizoctonia

Alternaria Aspergillus