Bacillus pumilus strain GHA 180 Page 1 of 27

Biopesticides Registration Action Document - DRAFT

 

BIOPESTICIDES REGISTRATION ACTION DOCUMENT

Bacillus pumilus strain GHA 180 

Pesticide Chemical (PC) Code: 016485

DRAFT DOCUMENT

U.S. Environmental Protection Agency

Office of Pesticide Programs

Biopesticides and Pollution Prevention Division

February 10, 2012
Bacillus pumilus strain GHA180 Page 2 of 27

Biopesticides Registration Action Document - DRAFT

TABLE OF CONTENTS

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

II. ACTIVE INGREDIENT OVERVIEW ................................................................................ 5

III. REGULATORY BACKGROUND................................................................................. 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 JUSTICE .......................................................................................... 13

VI. RISK MANAGEMENT DECISION ................................................................................. 13

VII. ACTIONS REQUIRED BY THE REGISTRANT ......................................................... 14

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS........................................... 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. MICROBIAL PESTICIDES DATA REQUIREMENTS............................ 20

APPENDIX B. PESTICIDE PRODUCTS............................................................................... 27
Bacillus pumilus strain GHA180 Page 3 of 27

Biopesticides Registration Action Document - DRAFT

BIOPESTICIDES 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

John 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 Regulatory Action Leader
Bacillus pumilus strain GHA180 Page 4 of 27

Biopesticides Registration Action Document - DRAFT

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 bacterium that produces metabolites that 

suppress the fungal plant pathogens Pythium ultimum, Fusarium oxysporum, Rhizoctonia solani

and Alternaria spp. This bacterium colonizes plant roots, forming a biofilm.

Premier Horticulture Ltd. has proposed to register three end-use pesticide products (EPs):

• GHA 180 Biological 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. (Contains 0.37 % Bacillus pumilus strain 

GHA180, a minimum of 1 x 109 viable spores/ml)

• Pro-Mix GHA180 and Pro-Mix Tandem. Both 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 contain Bacillus 

pumilus strain GHA180 at 0.001%, with a minimum of 5 x 104 viable spores per a gram 

of media.)

Pro-Mix GHA180 and Pro-Mix Tandem are each formulated into eight different custom blended 

growing mixes: BX, HP, TA, FPX, MPX, PGX, VFT and BRK Blends. There are16 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 the 

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 for 

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 product 

analysis data requirements for Bacillus pumilus strain GHA 180, including product chemistry 

and composition, analysis of samples, and physical and chemical characteristics.

Adequate mammalian 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 Bacillus 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. It 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 - DRAFT

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 

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 EPA believes there may be significant public interest.

 

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

pesticide products containing the new active ingredient, Bacillus pumilus 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 numbers associated with these registration actions and 

accessed through either http://www.regulations.gov/ or 

http://www.epa.gov/pesticides/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 labels for the end-use products, GHA 

180 Biological Fungicide (EPA File Symbol 74267-E , Pro-Mix ‘GHA180’(EPA File Symbol 

74267-G), and Pro-Mix Tandem (EPA File Symbol74267-U). While a final decision on 

registration is contingent upon review and consideration of public comments, EPA presently 

believes that, the risk assessment and information 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: International 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.,

use sites, application rates, methods of application, 

formulation types, and target pests) regarding

the registered pesticide products containing this 

active ingredient.

III. REGULATORY BACKGROUND
Bacillus pumilus strain GHA180 Page 6 of 27

Biopesticides Registration Action Document - DRAFT

A. Applications for Pesticide Registration

On February 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 74267–

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 receipt of 

these applications to register pesticide products containing a new active ingredient (75 Federal 

Register (FR) 71698) 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), Premier Horticulture 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 applied 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 

evaluated for this BRAD were considered in light of these requirements.

The classifications for each data submission are assigned by EPA science reviewers and indicate

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,” 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 often will state that the 

study is not required by 40 CFR Part 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 acute toxicity data requirements, Toxicity Categories are assigned based on the 

hazard(s) identified from studies and/or other information submitted to EPA in support of a 

pesticide registration. The active ingredient or particular 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 - DRAFT

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 bacterium produces metabolites that suppress the fungal plant 

pathogens Pythium ultimum, Fusarium oxysporum, Rhizoctonia solani and Alternaria spp. and 

colonizes plant roots, forming a biofilm. All product analysis data requirements for Bacillus 

pumilus strain 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 pumilus 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 strain GHA180 because of the lack of acute toxicity/pathogenicity in the Tier I 

studies.

For a comprehensive 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 Table 7 in Appendix A.

a. Acute Toxicity/Pathogenicity – Tier I

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

Guideline 885.3050; and Acute Pulmonary Toxicity/Pathogenicity (OCSPP Guideline 

885.3150;Master Record Identification Number (MRID No 48005019.):

Waiver rationale for acute oral and pulmonary toxicity/pathogenicity data requirements were 

submitted by Premier Horticulture Ltd, based on the acute injection toxicity and pathogenicity 

study and the oral, dermal and inhalation acute toxicity study results discussed below. All of 

those tests showed no evidence of infectivity, pathogenicity, toxicity, or other adverse effects. 

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

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

toxicity/pathogenicity data requirements for this active ingredient.

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 (1.4×1010 – 6.0×1010 CFU/mL) at a dose of 6.8×107 CFU/animal. Two control 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, and cecum contents of the rats given live 

bacteria on study days 0, 7, 14 and 21, and on day 21 for the control groups. All animals gained 

weight and appeared normal during the study, and no visible abnormalities were noted at 

necropsy. The test organism completely 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 rats 
Bacillus pumilus strain GHA180 Page 8 of 27

Biopesticides Registration Action Document - DRAFT

when dosed intravenously at 6.8×107 CFU/animal. This study was classified 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

were 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 (1.4×1010 – 6.0×1010 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 

LD50 > 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 (1.4×1010 –

6.0×1010 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 deformed 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 LD50 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

(1.4×1010 – 6.0×1010 CFU/mL) aerosol for 4 hours at a concentration of 2.18 mg/L, then observed for 14 

days. All animals gained weight and appeared normal throughout the study. No abnormalities were 

observed at necropsy. The LC50 > 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 (1.4x1010 – 6.0x1010 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. Bacillus pumilus GHA180 TGAI was mildly irritating to 

the eye. This study was classified as Acceptable; EPA Toxicity Category III.

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 (1.4×1010 – 6.0×1010 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 noted on 1 animal at 1 hour after patch removal; this cleared 
Bacillus pumilus strain GHA180 Page 9 of 27

Biopesticides Registration Action Document - DRAFT

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 Toxicology 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 

designate.” 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 where EPA 

will determine which, if any, of the Tier 2 tests are 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 and 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 

pesticide chemicals. Accordingly, EPA anticipates issuing future EDSP orders/data call-ins for 

all pesticide active ingredients. 

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: 

http://www.epa.gov/endo/.

2. Federal Food, Drug, and Cosmetic Act (FFDCA) 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 residue in or on a food) only if 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 include occupational exposure. Pursuant to section 408(c)(2)(B) 
Bacillus pumilus strain GHA180 Page 10 of 27

Biopesticides Registration Action Document - DRAFT

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 establishing 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 to pesticide 

residues. First, EPA determines the toxicity 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 section 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 this information to human risk. EPA has also considered available information 

concerning the variability of the sensitivities of major identifiable subgroups of consumers, 

including infants and children.

Based on the acute toxicity/pathogenicity data and information discussed previously and 

presented in Table 7 in Appendix A, the 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, lawns, 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 (9th 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 production 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 - DRAFT

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 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). 

Should this microbial pesticide be present in drinking water, 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, Residential Risk Characterization: The use sites for these products include 

greenhouse and contained nurseries. As stated previously, Bacillus pumilus is naturally present in 

many habitats. Based on the studies and other information submitted to satisfy data requirements 

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 tolerance 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 does not appear to produce a 

toxic metabolite as its mode of action against target pests. Therefore, for the purposes of this 

tolerance action, EPA has assumed that Bacillus pumilus strain GHA180 does 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 effects of 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 section 408(b)(2)(C) provides that 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 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 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 Table 

7 in Appendix A, EPA concludes that there is a reasonable certainty that no harm will result to 
Bacillus pumilus strain GHA180 Page 12 of 27

Biopesticides Registration Action Document - DRAFT

the U.S. population, including infants and children, from aggregate exposure to the residues of 

Bacillus pumilus strain GHA180. This 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 does not demonstrate toxic, 

pathogenic, 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 pumilus strain GHA180 is not expected to pose any undue risk. 

Appropriate 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 least 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 

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 when Bacillus pumilus strain GHA180 pesticide products are used in 

accordance with EPA-approved labeling.

C. Environmental Assessment (40 CFR § 158.2150)

The Agency has performed an environmental risk assessment for the proposed uses of Bacillus 

pumilus strain GHA180. The proposed use sites for Bacillus pumilus strain GHA180 were 

limited to greenhouse and contained nurseries. EPA determined that these use sites for Bacillus 

pumilus strain GHA180 would not result in significant nontarget species exposure and did 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 application 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 also 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 

increase 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.

Based on the data, literature citations, and data waiver rationale provided by the applicant, BPPD
Bacillus pumilus strain GHA180 Page 13 of 27

Biopesticides Registration Action Document - DRAFT

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

uses of Bacillus pumilus strain 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 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; (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 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 GHA180, 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 EPA’s web site 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 Bacillus pumilus strain 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 labels. 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 labels, as well as the data and 

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

will not cause any unreasonable adverse effects on the environment, and the GHA 180 
Bacillus pumilus strain GHA180 Page 14 of 27

Biopesticides Registration Action Document - DRAFT

Biological Fungicide, Pro-Mix ‘GHA180’, and Pro-Mix Tandem EPs (end-use products), in 

particular, are likely to protect against fungal pests as claimed, thereby satisfying criterion 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 ‘GHA180’, 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., outdoor or aquatic applications), EPA will likely require that 

additional data be submitted in conjunction with a registration amendment request.

VII. ACTIONS REQUIRED BY THE REGISTRANT

A. Final Printed Labeling

Before releasing pesticide products containing 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 sections, 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 all 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 under the 

provisions of 40 CFR § 158.2140(d).
Bacillus pumilus strain GHA180 Page 15 of 27

Biopesticides Registration Action Document - DRAFT

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 weight of animal

CFR Code of Federal Regulations

cfu colony-forming unit(s)

cfu/kg colony-forming units per kilogram

cfu/mL colony-forming units per milliliter

cP centipoise

DSMZ Deutsche Sammlung von Mikroorganismen und Zellkulturen (the German Strain 

Collection 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 Quality Protection Act

FR Federal Register

g/mL gram per milliliter

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 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

ULV ultra low volume
Bacillus pumilus strain GHA180 Page 16 of 27

Biopesticides Registration Action Document - DRAFT

IX. 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 pumilus 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 

Number: 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 Chemistry 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) Product 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.

48005010 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 prepared 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 - DRAFT

Unpublished 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 Pesticide 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. Project 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 Stillmeadow, Inc. 15 p.

48005022 Carter, L. (2009) Acute Inhalation Toxicity Study in Rats: Bacillus pumilus GHA180 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 GHA180 TGAI: 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-180 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 Biological 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 - DRAFT

.48537303 Libman, G. (2011) Microbiological Analyses: Certificates of Analysis 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. Unpublished 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 pumilus 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 Caused 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 species: 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. New Disease Reports 19:54.

From, C., V. Hormazabal and P.E. Granum. 2007. Food poisoning associated with pumilacidinproducing Bacillus pumilus in rice. Int. 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 einiger 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: clinical features, epidemics and outcomes. Int. J. Food Microbiol.

115(3):319-24.

Saleh, O.I., P.-I. Huang and J.-S. Huang. 1997. 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 - DRAFT

APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS 

(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 § 158.2120)

OCSPP 

Guideline 

Number 

Data Requirement

Results

MRID No.

TGAI GHA 180 (EP)

885.1100 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 applicable 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

885.1300 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 samples 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 Submitted data fulfill the 

requirement for an 

enforcement analytical 

method

48005001
Bacillus pumilus strain GHA180 Page 21 of 27

Biopesticides Registration Action Document - DRAFT

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 

Guideline 

Number 

Data Requirement

Results

MRID No.

TGAI GHA 180 Biological Fungicide

(EP)

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 

Elevated

Temperatures, Metals,

and Metal Ions

Vegetative cells expected to denature at >50oC. 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

48559502
Bacillus pumilus strain GHA180 Page 22 of 27

Biopesticides Registration Action Document - DRAFT

TABLE 3. Product Analysis Data Requirements for the End-Use Product (EP), Pro-Mix GHA 180 (40 CFR § 

158.2120)

OCSPP Guideline 

Number

Data Requirement Results MRID No.

885.1100 Product Identity Submitted data fulfill the requirement for 

product identity.

48005003-

48005010

885.1200 Manufacturing Process Submitted data fulfill the requirement for 

manufacturing process.

48005003-

48005010

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 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 statement 

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 requirement 

of a tolerance without any numerical limitation.

48005003-

48005010
Bacillus pumilus strain GHA180 Page 23 of 27

Biopesticides Registration Action Document - DRAFT

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.

885.1100 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 current 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 +30o

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.

830.6317 Storage Stability Storage stability was shown up to two 

years in peat growing mixes stored 

outdoors with temperature variations 

from -30 to +30o

C.

48005011-

48005018

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/ft3 CSFs dated 

1/7/11
Bacillus pumilus strain GHA180 Page 24 of 27

Biopesticides Registration Action Document - DRAFT

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 was shown up to two 

years in peat growing mixes stored 

outdoors with temperature variations 

from -30 to +30oC.

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/ft3 CSFs dated 

1/7/11
Bacillus pumilus strain GHA180 Page 25 of 27

Biopesticides Registration Action Document - DRAFT

TABLE 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

TGAI 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 

submitted 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 Acute 

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 (1.4×1010 – 6.0×1010 CFU/mL) at a dose of 6.8×107

CFU/animal and observed up to 21 days. All animals 

survived, gained weight, and appeared normal during the 

study. No observable abnormalities were noted in any 

animal at necropsy. The test organism completely cleared 

from the blood, kidneys, mandibular lymph nodes and 

cecum contents of the animals by day 14, and showed 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 6.8×107 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 LD50 was >5000 mg/kg bw.

Classification: Acceptable

TOXICITY CATEGORY IV

Data on 

TGAI supports these EPs

48005020

870.1200 Acute Dermal 

Toxicity

The LD50 was >5050 mg/kg bw.

Classification: Acceptable

TOXICITY CATEGORY IV

Data on 

TGAI supports these EPs

48005021

870.1300 Acute 

Inhalation 

Toxicity

The LC50 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 treated 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

(1.4×1010 – 6.0×1010 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

TOXICITY CATEGORY IV

Data on 

TGAI supports these EPs

48005024
Bacillus pumilus strain GHA 180 Page 26 of 27

Biopesticides Registration Action Document - DRAFT

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. 

The current label use patterns for 

greenhouse and nursery use would not 

result in significant nontarget species 

exposure.

48005019

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 

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 GHA180 based on the current uses and application methods.
Bacillus pumilus strain GHA 180 Page 27 of 27

Biopesticides Registration Action Document - DRAFT

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% End-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

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