EPA BIOPESTICIDES AND POLLUTION PREVENTION DIVISION

NOTICE OF FILING TEMPLATE FOR PESTICIDE PETITIONS PUBLISHED IN THE

FEDERAL REGISTER 

EPA Biopesticides and Pollution Prevention Division contact: Susanne Cerrelli at (703) 308-8077

EPA has received a pesticide petition [insert petition number] from Certis USA LLC, 9145 Guilford 

Road, Suite 175, Columbia, MD 21046 proposing pursuant to section 408(d) of the Federal Food, 

Drug, and Cosmetic Act (FFDCA), 21 U.S.C. 346a(d), to amend 40 CFR part 180 to establish an 

exemption from the requirement of a tolerance for residues of the microbial pesticide Bacillus 

mycoides isolate J in or on all agricultural commodities.

Pursuant to section 408(d)(2)(A)(i) of the FFDCA, as amended, Certis USA LLC has submitted the

following summary of information, data, and arguments in support of their pesticide petition. This 

summary was prepared by Certis USA LLC and EPA has not fully evaluated the merits of the 

pesticide petition. The summary may have been edited by EPA if the terminology used was unclear, 

the summary contained extraneous material, or the summary unintentionally made the reader 

conclude that the findings reflected EPA’s position and not the position of the petitioner. 

I. Certis USA LLC Petition Summary

Petition number 4F8252

A. Product name and Proposed Use Practices 

The microbial pest control agent is Bacillus mycoides isolate J. Bacillus mycoides isolate J is a 

naturally occurring, spore-forming microorganism, one of six species that comprise the Bacillus

cereus group. It was isolated in a field of infected sugar beets. Bacillus mycoides isolate J is a 

common, ubiquitous soil bacterium that is found worldwide. The mode of action of Bacillus 

mycoides isolate J is to induce systemic resistance, triggering the plant’s defenses to control or 

suppress plant diseases.

Application of live vegetative cells of Bacillus mycoides isolate J to the leaves of growing crops 

induces systemic resistance in the plant that is characterized by an oxidative burst and production of 

pathogenesis related proteins including chitinase and peroxide and beta glucanase, but does not 

cause plant death or tissue necrosis.
B. Product Identity/Chemistry 

1. Identity of the pesticide and corresponding residues. 

The microbial pest control agent Bacillus mycoides isolate J is a naturally occurring soil organism 

which will induce systemic resistance to certain plant diseases. Under proper environmental 

conditions, Bacillus mycoides isolate J induces systemic resistance in a broad range of plants and 

will be effective in controlling fungal and bacterial diseases such as sugar beets, cucurbits, tomatoes, 

ornamentals, solanaceous plants and other crops.

Bacillus mycoides isolate is a common and ubiquitous soil microorganism that was first isolated by

the Jacobson group from leaves of sugar beets grown in Montana in 1994. Bacillus mycoides isolate 

J forms very distince spiral colonies on nutrient agar, and is one of six species that comprise the 

B.cereus group. A number of DNA sequence matching techniques have been used to distinguish

BmJ from other B.cereus group isolates; this species is genetically separate from other species in the 

group.

The active ingredient can be cultured readily on various culture media and is cultured by

conventional submerged fermentation. Spores of the fermentation are recovered from the 

fermentation broth and dried to create the technical grade active ingredient. A master stock is 

isolated from a single colony isolate by plating on potato dextrose agar. The culture has been 

deposited in the Agricultural Research Service depository in Peoria, IL with the accession number 

NRRL B-30890.]
2. Magnitude of residue at the time of harvest and method used to determine the residue. 

Bacillus mycoides isolate J is a naturally occurring soil microorganism which has been isolated in 

agricultural soils worldwide and, as such, long term exposure to humans, in crops is expected to 

occur. No toxicity or pathogenicity to humans has been reported in the literature or from tests,

resulting from exposure to Bacillus mycoides isolate J, and this microorganism is not known to be 

toxic or pathogenic or toxic to any environmental animal or plant species. Therefore, when used 

according to label directions, Bacillus mycoides isolate J would not result in residues that are of 

toxicological concern. 

The spores of Bacillus mycoides isolate J are affected by sunlight, temperature, humidity, substrate, 

and chemicals. However, it is the systemic induction of resistance to certain fungal and bacterial 

diseases by the plant that provides control, and not the infectivity of germinating spores. The halflife of the spores is relatively short, and normal washing after harvest and before sale would further 

reduce or remove any residues. 

3. A statement of why an analytical method for detecting and measuring the levels of the pesticide 

residue are not needed. 

The acute toxicity studies discussed in Section C of this document are sufficient to show that there 

are no foreseeable human or domestic health hazards likely to arise from the use of the product as 

proposed. This soil microorganism is present naturally and Certis USA is requesting an exemption 

from the requirement for a tolerance for any residues remaining in/on all agricultural commodities. 

Since enforcement of residue levels would not be needed, an enforcement analytical method is not 

required.

C. Mammalian Toxicological Profile 

Acute oral toxicity/pathogenicity- rat –To evaluate the acute oral toxicity, infectivity/pathogenicity 

of Bacillus mycoides isolate J Technical Grade Active Ingredient, a single high dose of 6.3 x 109

CFU per gram or 6.3 x 108 was administered by gavage to rats. At necropsy, no internal observable 
abnormalities were noted, no signs of toxicological effects. There was no mortality during the study 

and the test material had cleared from brain, mesenteric lymph nodes, blood, kidney, spleen, liver,

lung and caecum 

Acute intravenous toxicity/pathogenicity – rat – the test substance was administered at a single high 

dose of 5.3x 108CFU/gram totwo groups or rats. No signs of toxicity or pathogenicity were obse

rved under the condition of the test. No abnormalities were observed for the duration of the study. 

The test material cleared completely from the brain, blood, lungs, kidneys, and cecum. A pattern of

clearance was established in the liver, 

Acute pulmonary toxicity/pathogenicity- rat –A single intratracheal dose 1.1 x 108 CFU/ rat was 

administered to test animals. No deaths occurred and there were no signs of toxicity observed in any 

animal under the conditions of this test. Although total clearance of the microorganism was not 

established, no clinical signs of toxicity or infectivity were noted for the duration of the study.

Acute oral toxicity –rat- the test substance (BmJ WG) at 5000 mg/kg was administered via gavage to 

female rats. No mortality occurred during the study, and no clinical signs of toxicity were observed. 

The acute LD50 was determined to be greater than 5,000 mg/kg. Based on the test results, EPA has 

classified the product at toxicity category IV.

Primary eye irritation – the test substance (BmJ WG) was instilled in the eye of albino rabbits and 

examined at 1, 24, 48 and 72 hours after treatment. The test substance (produced irritation which

was reversible within 48 hours after treatment. No irritation was observed in any eyes at 72 hours.

Based on the test results EPA has classified the product as toxicity category III.

Dermal irritation – rabbit – in a four hour dermal exposure to the skin of rabbits, the test substance 

(BmJ WG) was non-irritating with a primary irritation index of zero. Based on the test results, EPA 

has classified the product as toxicity category IV.

Acute dermal toxicity - rats – the test substance (BmJ WG) was administered as a single dose at 

5050 mg/kg. No mortality was observed for the duration of the study, and there were not clinical 

signs of toxicity at any time during the study. Based on the test results, the LD50 for BmJ WG is 

greater than 5050 mg.kg.

D. Aggregate Exposure 

1. Dietary exposure. 

i. food

Bacillus mycoides isolate J is a naturally occurring soil microorganism, which has been isolated in 

soils worldwide and as such, humans, animals, and plants are constantly exposed to this

microorganism. It is expected that, when used as proposed, Bacillus mycoides isolate J would not 

result in residues that are of toxicological concern. Bacillus mycoides isolate is sensitive to 
environmental factors such as sunlight and temperature. Bacillus mycoides isolate J spores lose their 

viability when exposed to periods of sunlight and temperatures consistent with mammalian body 

temperatures. After harvest any remaining spores on crop leaves will be exposed to unfavorable 

conditions (e.g. dryness), and are not likely to germinate; any potentially occurring residual deposits 

on these crops will not harm humans because Bacillus mycoides isolate J demonstrates no toxicity, 

infectivity, or pathogenicity in appropriate tests. Further the optimal temperature range for, 

germination between 27 and 35 degrees C while the normal body temperatures of humans, other 

mammalian species and avian species is significantly higher than 35 degrees C and therefore 

unsuitable for germination of Bacillus mycoides isolate J.

While the proposed use pattern may result in dietary exposure with possible residues in or on certain 

agricultural commodities, negligible, to no risk, is expected for the general population, including 

infants and children, or animals because Bacillus mycoides isolate J demonstrated no pathogenicity 

or toxicity at the maximum doses tested.

Since Bacillus mycoides isolate J is a microorganism that is ubiquitous in soils, humans may be 

exposed dermally and orally to this common fungus when they get soil on their hands or clothing, or 

handle pets that have played in soil. Importantly, no toxicological endpoints were identified for

Bacillus mycoides isolate J, and there is no evidence of adverse effects from oral, dermal, or 

pulmonary exposure to this MPCA. It is not expected that the pesticidal use of Bacillus mycoides

isolate J will significantly increase the non-dietary and non-occupational exposure to the fungus. 

 

Based on the proposed use patterns, in which Bacillus mycoides isolate J is applied directly to 

foliage of agricultural and ornamental crops, the potential for non-dietary, non-occupational

exposures to Bacillus mycoides isolate J pesticide residues by the general population, including 

infants and children, is low. Moreover, even in the unlikely event of non-dietary, non-occupational 

exposures to Bacillus mycoides isolate J residues, no harm is expected because no toxicity or 

pathogenicity was found in mammalian studies that included high levels of oral, pulmonary, and 

dermal exposure.

. Drinking water. 

Bacillus mycoides isolate J is not known as an aquatic microorganism, and therefore is no evidence 

in the literature that this microorganism will germinate in aquatic habitats. There are no aquatic use 

sites permitted for this pesticide, so exposure to drinking water is not expected. Accordingly, the use 

of this MCPA on terrestrial plants is not anticipated to lower the quality of drinking water. Even if 

low levels of the microbe were present in drinking water, no risk to the general public would be

expected because Bacillus mycoides isolate J demonstrated no pathogenicity or toxicity at the 

maximum dose tested. The potential of significant transfer to drinking water is minimal to

nonexistent. 

2. Non-dietary exposure.
Since Bacillus mycoides isolate J is a naturally occurring microorganism, there may be non-dietary

exposure, but not above background levels of this naturally occurring microorganism. 

E. Cumulative Exposure 

There is reasonable certainty that no harm will result to the U.S. population, including infants and 

children, from aggregate exposures to residues of Bacillus mycoides isolate J as a result of its 

proposed uses. This includes all anticipated dietary exposures and all other exposures for which

there is reliable information. There appears to be no potential for harm, from this microorganism in 

its use as a microbial pesticide in agricultural commodities. Furthermore, the organism is naturally 

occurring in soils, and is non-toxic and non-pathogenic to animals and humans. This conclusion is 

based on the results of the mammalian toxicity tests referenced in Section C. Moreover, potential 

non-occupational inhalation or dermal exposure is not expected to pose any adverse effects to

exposed populations via aggregate and cumulative exposure. 

Therefore no cumulative effects from the residues of this product with other related microbial 

pesticides are anticipated.

F. Determination of Safety for U.S. population, Infants and Children

There is reasonable certainty that no harm will result to the U.S. population, including infants and 

children, from aggregate exposures to residues of Bacillus mycoides isolate J as a result of its 

proposed uses. This includes all anticipated dietary exposures and all other exposures for which 

there is reliable information. As discussed previously, there appears to be no potential for harm, 

from this microorganism in its use as a systemic inducer of resistance by the treated plant/crop

Furthermore, the organism is naturally occurring in the environment, and non-toxic and nonpathogenic to animals and humans. This conclusion is based on the results of the mammalian 

toxicity tests discussed in Section C. Moreover, potential non-occupational inhalation or dermal 

exposure is not expected to pose any adverse effects to exposed populations via aggregate and 

cumulative exposure.

G. Effects on the Immune and Endocrine Systems 

There is no information to indicate that Bacillus mycoides isolate J would have an effect in humans 

similar to an effect produced by a naturally-occurring estrogen or other endocrine effects. There is 

no known metabolite produced by this microorganism that acts as an endocrine disruptor. The 

submitted toxicity/pathogenicity studies in rodents indicate that following dosing no viable spores 

were found in organs or tissues of test animals. It is concluded that there will be no incremental 

adverse effects to the endocrine system.

H. Existing Tolerances 
Under 40 CFR 180.1269, there is only a temporary exemption from the requirement of a tolerance

for Bacillus mycoides isolate J when used as a fungicide on potatoes in accordance with a valid 

Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA) section 18 emergency exemption

established. This temporary exemption from the requirement of a tolerance expires and is revoked on 

December 31, 2015.

There is no tolerance or other exemption from the requirement of a tolerance established for residues 

of Bacillus mycoides isolate J.

I. International Tolerances 

The European Food Safety Authority Scientific Report (2007), 115, 1-161, determined that under 

Regulation (EC) No 396/2005 of the European Parliament and of the Council on Maximum

Residue Levels for pesticides (28 October 2007) that no MRLs and no residue definitions have

to be established for the use of Bacillus mycoides isolate J on agricultural crops.

There is no CODEX maximum residue level for residues of Bacillus mycoides isolate J.