EPA BIOPESTICIDES AND POLLUTION PREVENTION DIVISION COMPANY NOTICE OF FILING FOR PESTICIDE PETITIONS PUBLISHED IN THE FEDERAL REGISTER  

EPA Biopesticides and Pollution Prevention Division contact:
Jeannine Kausch 703-347-8920

SUBMISSION: Email the completed template to: hollis.linda@epa.gov.

TEMPLATE:
AFS32321 Crop Protection, Inc.
Petition number: 9F8816

	EPA has received a pesticide petition (9F8816) from AFS32321 Crop Protection, Inc. 104 T.W. Alexander Dr. Research Triangle Park, NC 27709 requesting, 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 microbial pesticide Bacillus subtilis strain AFS032321 (NRRL B-67089). Pursuant to section 408(d)(2)(A)(i) of FFDCA, as amended, AFS32321 Crop Protection, Inc. has submitted the following summary of information, data, and arguments in support of their pesticide petition. This summary was prepared by AFS32321 Crop Protection, Inc. 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. AFS32321 Crop Protection, Inc. Petition Summary

	9F8816

A. Product Name and Proposed Use Practices

AFS032321 Technical
Theia[TM] Fungicide

AFS032321Technical is a technical grade active ingredient (TGAI)/manufacturing use product (MUP) proposed for use in one end use product: THEIA(TM) Fungicide. 

The end use product is broad spectrum fungicidal and bactericidal biological products for the control or suppression of many important foliar and soil borne plant diseases.

B. Product Identity/Chemistry

1. Identity of the pesticide and corresponding residues. Bacillus subtilis strain AFS032321 (CAS No. Not applicable): Bacillus subtilis is ubiquitous in the environment, occurs naturally in soils, and has been identified in association with plant roots, organic material and inorganic material in soil samples.

2. Magnitude of residues at the time of harvest and method used to determine the residue. It is expected that, when used as proposed, Bacillus subtilis strain AFS032321 would not result in residues that are of toxicological concern. 

3. A statement of why an analytical method of detecting and measuring the levels of the pesticide residue are not needed. An analytical method for residues is not applicable. It is expected that, when used as proposed, Bacillus subtilis strain AFS032321 would not result in residues that are of toxicological concern

C. Mammalian Toxicological Profile	Studies to evaluate the safety to mammals were conducted on the technical grade active ingredient (TGAI) and are summarized below.

 Acute Oral Toxicity/Pathogenicity (OCSPP 885.3050): An acute oral toxicity/pathogenicity study was conducted in Sprague-Dawley CDR rats to determine the toxicity and pathogenicity of AFS032321 TGAI. Thirty-two (32) healthy rats (16 males and 16 females) were selected for the test and distributed into three main study groups (one treatment group and two controls). Control animals were left untreated.

Fecal samples from the treatment group animals contained an average of 8.5 x 10[7] (males) and 5.1 x 10[7] (females) CFU/g viable AFS032321 TGAI colonies on Day 2. The samples fell below the limit of detection by Day 7 (males) and Day 14 (females). 

Concentrations in blood, brain, kidney, liver, mandibular and mesenteric lymph node and spleen samples were below established limits of detection throughout the study in treated males. In treated females, AFS032321 TGAI colonies were detected, on Day 3, in kidney, lymph nodes and blood samples. Subsequent samples collected on and after Day 7 showed concentrations below the limit of detection.

Under the conditions of this study, the acute oral administration of AFS032321 TGAI, at a dose level of 5.4 x 10[8] viable CFU by oral gavage produced no evidence of acute toxicity or pathogenicity in Sprague-Dawley rats. (MRID No. 509861-04) 

 Acute Pulmonary Toxicity/Pathogenicity (OCSPP 885.3150): An acute pulmonary toxicity/pathogenicity study was conducted in Sprague-Dawley CDR rats to determine the toxicity and pathogenicity of AFS032321 TGAI.  Thirty-eight (38) healthy rats (19 males and 19 females) were selected for the test and distributed into two main study groups.  Control animals were left untreated.

At study termination, complete clearance was achieved for blood, brain, kidney and cecum contents. Partial clearance was achieved for liver, spleen, lung and lymph nodes.  Liver and spleen counts were close to limit of detection. Lung and lymph nodes had a twofold and a threefold decrease indicating a start of clearance of the organism.

Under the conditions of the study, the acute pulmonary administration of AFS032321 TGAI, at a dose level of 5.89 x 108 viable CFU produced no evidence of acute toxicity in Sprague-Dawley rats. The trend of decrease in counts was not sufficient to establish a pattern of clearance but it is possible that given more time such a pattern may have emerged. Although not fully cleared from lung and lymph node, overall there was 12-fold decrease for lung and a 52-fold for lymph nodes indicating a pattern of clearance of the organism. Given a longer observation period, it is possible that full clearance could be achieved in these tissues, although this was not demonstrated in this particular study. This trend of decrease in counts along with the absence of any toxicological or pathological findings suggests that AFS032321 TGAI administered to rats in a single intra-tracheal dose containing 5.89x108 CFU is not toxic or pathogenic. (MRID No. 509861-05) 

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Primary Eye Irritation (OCSPP 870.2400): One hour after test substance instillation, minimal conjunctivitis was noted for all three treated eyes. There was no corneal opacity or iritis observed in any treated eye during this study. The overall incidence and severity of irritation decreased with time. All animals were free of ocular irritation by 48 hours. The Maximum Mean Total Score of AFS032321-WP also known as AFS032321 Technical is 6.7. Under the conditions of this study, AFS032321-WP is classified as minimally irritating to the eye. (MRID No. 509861-06)
 
 Primary Dermal Irritation (OCSPP 870.2500): Within 30-60 minutes of patch removal, all three treated sites exhibited very slight to well-defined erythema and very slight to slight edema. The overall incidence and severity of irritation decreased gradually with time. All animals were free of dermal irritation by Day 7 (study termination). The Primary Dermal Irritation Index for AFS032321-WP also known as AFS032321 Technical is 2.6. Under the conditions of this study, AFS032321-WP is classified as moderately irritating to the skin. (MRID No. 509861-07)

 Hypersensitivity Incidents (OCSPP 885.3400): The registrant has noted that no incidents of hypersensitivity or any other adverse effects have occurred through the research, development or testing of the active ingredient and its related end-use product. Should any incidents occur, they will be reported per FIFRA Section 6(a)(2).

The results of toxicity testing show there is no risk to human health from the active ingredient. Bacillus subtilis strain AFS032321 is not toxic, pathogenic, infective or irritating to mammals.

D. Aggregate Exposure

	1. Dietary exposure.

i. Food. Dietary exposure from use of Bacillus subtilis strain AFS032321, as proposed, is expected to at low levels. The risk posed to adults, infants and children is minimal because of the low acute oral toxicity/pathogenicity potential of the microbial pesticide. In addition, standard practices of washing, peeling, cooking or processing fruits and vegetables reduces residues of Bacillus subtilis strain AFS032321 and further minimizes dietary exposure.

The results of toxicity testing indicate there is no risk to human health or the environment from Bacillus subtilis strain AFS032321. There are no reports of ecological or human health hazards caused Bacillus subtilis strain AFS032321. It does not produce recognized toxins, enzymes, or virulence factors normally associated with mammalian invasiveness or toxicity. The absence of acute toxicity or pathogenicity in laboratory animals demonstrates the benign nature of this strain.

ii. Drinking water. Similarly, exposure to humans from residues of Bacillus subtilis strain AFS032321 in consumed drinking water would be unlikely. Potential exposure to surface water would be negligible and exposure to drinking water (well or groundwater) would be impossible to measure. Bacillus subtilis is ubiquitous in the environment and has been identified in soils and waters worldwide.

The intended use of Bacillus subtilis strain AFS032321 is for the purposes of disease control. The risk of the microorganism passing through the soil to groundwater is minimal. Bacillus subtilis is ubiquitous in the environment, occurs naturally in soils, and has been identified in association with plant roots, organic material and inorganic material in soil samples. Additionally, the bacterium would not tolerate the conditions water is subjected to in a drinking -water facility (including: chlorination, pH adjustments, high temperatures and/or anaerobic conditions).

The results of toxicity testing indicate there is no risk to human health or the environment from Bacillus subtilis strain AFS032321. There are no reports of ecological or human health hazards caused by this microorganism. It does not produce recognized toxins, enzymes, or virulence factors normally associated with mammalian invasiveness or toxicity. The absence of acute toxicity or pathogenicity in laboratory animals demonstrates the benign nature of this strain.

2. Non-dietary exposure. The potential for non-dietary exposure to the general population, including infants and children, is unlikely as the proposed use sites are agricultural. The intended use of Bacillus subtilis strain AFS032321 is for the purposes of disease control. Personal Protective Equipment (PPE) mitigates the potential for exposure to applicators and handlers of the proposed products, when used in agricultural settings.

The lack of acute oral toxicity, infectivity, and pathogenicity support an exemption from the requirement of a tolerance for Bacillus subtilis strain AFS032321. There are no reports of ecological or human health hazards caused by Bacillus subtilis. It is not known to produce recognized toxins, enzymes, or virulence factors normally associated with mammalian invasiveness or toxicity. The absence of acute toxicity or pathogenicity in laboratory animals demonstrates the benign nature of this strain. Non-dietary exposures would not be expected to pose any quantifiable risk due to a lack of residues of toxicological concern.

E. Cumulative Effects

It is not expected that, when used as proposed, Bacillus subtilis strain AFS032321 would result in residues that are of toxicological concern. The intended use of Bacillus subtilis strain AFS032321 is for the purposes of disease control. The results of toxicity testing indicate there is no risk to human health or the environment from Bacillus subtilis strain AFS032321. There are no reports of ecological or human health hazards caused by this microorganism It does not produce recognized toxins, enzymes, or virulence factors normally associated with mammalian invasiveness or toxicity. The absence of acute toxicity or pathogenicity in laboratory animals demonstrates the benign nature of this strain.

F. Safety Determination

1. U.S. population. It is not expected that, when used as proposed, Bacillus subtilis strain AFS032321 would result in residues that are of toxicological concern. The intended use of Bacillus subtilis strain AFS032321 for the purpose of disease control. The results of toxicity testing indicate there is no risk to human health or the environment from Bacillus subtilis strain AFS032321. There are no reports of ecological or human health hazards caused by this microorganism It is not known to produce recognized toxins, enzymes, or virulence factors normally associated with mammalian invasiveness or toxicity. The absence of acute toxicity or pathogenicity in laboratory animals demonstrates the benign nature of this strain. There is a reasonable certainty of no harm to the general US population from exposure to this ingredient. 

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2. Infants and children. As mentioned above, it is not expected that, when used as proposed, Bacillus subtilis strain AFS032321 would result in residues that are of toxicological concern. There is a reasonable certainty of no harm for infants and children from exposure to Bacillus subtilis strain AFS032321 from the proposed uses.
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G. Effects on the Immune and Endocrine Systems

To date there is no evidence to suggest that Bacillus subtilis strain AFS032321 functions in a manner similar to any known hormone, or that it acts as an endocrine disruptor.

H. Existing Tolerances

There is no US EPA tolerance or tolerance exemption for Bacillus subtilis strain AFS032321.

I. International Tolerances

A Codex Alimentarium Commission Maximum Residue Level (MRL) is not established for Bacillus subtilis strain AFS032321.


