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 pe sticide 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 establ ish 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

o f information, data, and arguments in support of their pesticide petition. This summary w as 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

u sed was unclear, the summary

c ontained 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 **Petitio n**  **number 4F**  **8252** A. Product name and Proposed Use Practices The microbial pest control agent is Bacillus mycoides isolate J

. Bacillus mycoides isolate J is a naturally o ccurring, 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 producti on 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 isolate d by the Jacobs on 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 te chniques 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 i solate by

p lating on potato dextrose agar. The culture has been deposited in the Agricultural Research Service depository in Peoria, IL with the accession number NRRLB

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

.

Bacillus mycoides isolate J is a naturally o ccurring 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

o r 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 half

life 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 likel y 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 anal ytical method is not required. C. Mammalian Toxicological Profile Acute oral toxicity

/pathogenicity

- rat

–To evaluate the acute oral toxicity, inf ectivity /pathogenicity of Bacillus mycoides isolate J Technical Grade Active Ingredient , a single high dose of 6.3 x 10 9 CFU per gram or 6.3 x 10 8 was administered by g avage to rats. At necropsy, no internal observableabnormalities were noted, no signs o f toxicological effects. There was no mortality during the study

and the test material had cleared from brain, mesenteric ly mph 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 10 8 CFU/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 comp letely from the brain, blood, lungs, kidneys, and cecum. Ap attern of clearance was established in the liver, Acute pulmonary

toxicity /pathogenicity

- rat

–A single intratracheal dose 1.1 x 10 8 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 LD 50 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 e ye 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. B ased on the test results, EPA has classified the product as toxicity category IV. Acute dermal toxicity

- rats

– the te st substance (BmJ WG) was administered as a single dose at 5050 mg/kg. No mortality w as 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 LD 50 for BmJ WG is greater than 5050 mg.kg. D. Aggregate Exposure 1. Dietary exposure

. i. food Bacillus mycoides isolate J is a naturally o ccurring soil microorganism , which has been isolated in soils worldwide and as such, humans, animals, and plants are constantly expo sed to this microorganism. Iti s 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 toenvironmental 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 . dry

ness), and are not likely

to germinate ; any

p otentially

oc curring 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 2 7 and 3 5 degrees C while the normal body temperatures of humans, other mammalian species and avian species is significantly higher than 3 5 degrees C and therefore unsuitable for germination of Bacillus mycoid es 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 becau se Bacillus m ycoides isolate J demonstrated no p athogenicity or toxicity

at the maximum dose s tested

. Since Bacillus mycoides isolate J is a microorganism that is ubiquitous in soils, humans may b e 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- o ccupational exposures to Bacillus mycoide s isolate J residues, no harm is expected because no toxicity or pathogenicity

w as 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 o f drinking water. Even if low levels of the microbe were present in drinking water, no risk to the g eneral 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 Ba cillus mycoides isolate J is a naturally occurring microorganism , there may b e 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 relia

ble 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. A s 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 th e 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. M oreover, potential non- occupational inhalation or dermal exposure is not expected to p ose 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 n aturally- 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 s ystem. H. Existing TolerancesUnder 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 MRL s 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 residue s of Bacillus mycoides isolate J.