### EPA BIOPESTICIDES AND POLLUTI ON PREVENTION DIVISION

COMPANY NOTICE OF FILING FOR PE STICIDE PETITIONS PUBLISHED IN

THE FEDERAL REGISTER

EPA Biopesticides and Pollution Prevention Division contact: ShaRon Carlisle; (703) 308-6427

SUBMISSION: E-mail the completed temp late to: hollis.linda@epa.gov.

TEMPLATE:

AFS009 Plant Protection, Inc. (EPA Co. No. 91197)

5F8410

EPA has received a pesticide petition ( 5F8410) from AFS009 Plant Protection, Inc. (EPA Co. No. 91197)

, 104 T.W. Alexander Drive, Building 18, Research Triangle Park, NC 27709 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 microbial pesticide
Pseudomonas chlororaphis subsp. aurantiaca strain AFS009



### .

Pursuant to section 408(d)(2)( A)(i) of FFDCA, as amended, AFS009 Plant Protection, Inc. (EPA Co. No. 91197) has submitted the following summary of information, data, and arguments in support of their pe sticide petition. This summary was prepared by AFS009 Plant Protection, Inc. (EPA Co. No. 91197) 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 read er conclude that the findings reflected EPA’s position and not the pos ition of the petitioner.

I. AFS009 Plant Protection, Inc. (EPA Co. No. 91197) Petition Summary

5F8410

A. Product Name and Proposed Use Practices

Product Name: HowlerTM Technical (100% ai; TGAI) – EPA File Symbol

91197-R HowlerTM (50% ai; EP) – EPA File Symbol 91197-G

Proposed Use Practice: HowlerTM Technical is a 100% ai Technical Grade Active Ingredient (TGAI) of Pseudomonas chlororaphis subsp. aurantiaca strain2

AFS009 and is proposed for manufacturing use only, for further formulation into registered end-use products.

HowlerTM is a 50% ai is formulated end-use product for use on growing plants and crops to control plant diseases including Rhizoctonia, Pythium, Fusarium, Phytophthora and Botrytis . HowlerTM may be mixed with water and applied as a foliar spray, soil drench, in furrow spray, transplant spray or dip, cuttings or bare root dip, hydroponic or chemigation application in greenhouse, agricultural field, turf and ornamental and home and garden use sites.



### B. Product Identity/Chemistry

1. Identity of the pesticid e and corresponding residues. Pseudomonas chlororaphis subsp. aurantiaca strain AFS009 (CAS No. Not applicable).

Pseudomonas chlororaphis is a common bacterium identified primarily in the soil. Pseudomonas chlororaphis subsp. aurantiaca is a Gram-negative, rod-shaped bacteria with one or more flagella for motility. Information regarding the name, identity and composition has been submitted to EPA and can be found in MRID No. 495680-01.

Like other Pseudomonas chlororaphis strains Pseudomonas chlororaphis subsp.

aurantiaca strain AFS009 is a plant-colonizing bacteria which controls fungal diseases by several modes of action, including competition and production of a variety of metabolites which are inhibitory to the fungi. (MRID No. 495680-01).



### 2. Magnitude of residues at the time of har vest and method used to determine the

residue. NA-Remove

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, Pseudomonas chlororaphis subsp. aurantiaca strain AFS009, 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 as follows:

1. Acute Oral Toxicity/Pathogenicity Study in Rats (OCSPP 885.3050):

HowlerTM Technical ( 100% Pseudomonas chlororaphis subsp. aurantiaca strain

AFS009 ) was not toxic or pathogenic in rats following acute oral exposure to a

concentrate of 3.73 x 10 9 (MRID No. 495680-02). The MPCA test substance and

the inactivated test substance were admin istered to rats by gavage in single high doses. The animals were observed frequently on the day of dosing (Day 0) for mortality and clinical signs of toxicity, and once daily thereafter for 21 days. An untreated control group was conducted concurrently. Tissue samples from treated rats were enumerated on days 0, 3, 7, 14, and 21. The test organism was not observed in plated blood, brain or liver. The urine from treated rats was completely clear of MPCA growth by 72 hours. The test organism cleared3

completely from the treated lungs, spleen, and kidney by Day 14. The mesenteric lymph nodes from treated rats did not achieve complete clearance, but declined to very low levels of growth found only in a single animal by Day 21. There were no signs of pharmacologic and/or toxicologic effects observed in any animal during the study, and no mortality occurred. The gross necropsy conducted at termination of the study revealed no internal abnormalities. The test substance, HowlerTM Technical, was determined to be non-toxic to rats and demonstrate a pattern of clearance when administered by oral gavage in a single dose of 3.73 x

10 9 CFU/rat.

2

. Acute Oral Toxicity (OCSPP 870.1100): The TGAI, HowlerTM Technical (100% Pseudomonas chlororaphis subsp. aurantiaca strain AFS009), was not toxic following acute exposure by the oral r oute. An acute oral toxicity study was conducted on rats using the up-and-down procedure to determine the potential for HowlerTM Technical to produce toxicity from a single oral dose (MRID No. 495680-03). An initial dose of 5,000 mg per kg body weight of the test substance was administered to a single female rat by gavage. This first rat survived and so two additional rats received a gavage dose of 5,000 mg/kg bw of the test material. All three animals survived, so no additional animals were tested. The rats were observed for mortality, clinical signs of toxicity and behavioral changes daily for 14 days. Body weights were recorded prior to exposure and again on Days 7 and 14. All animals survived test substance administration, gained body weight, and appeared active and healthy during the study. Necropsies were performed on all animals at terminal sacrifice. Under the study conditions, the acute oral LD50 of HowlerTM Technical was greater than 5,000 mg/kg body weight in female rats (Toxicity Category IV).

3. Acute Dermal Toxicity (OCSPP 870.1200): The TGAI, HowlerTM Technical (100% Pseudomonas chlororaphis subsp. aurantiaca strain AFS009) was not toxic following acute exposure by the dermal route. An acute dermal toxicity study was conducted on rats to determine the potential for Howler Technical to produce toxicity from a single topical application (MRID No. 495680- 04). Five thousand milligrams (mg) of the test substance per kilogram (kg) of body weight was applied to the skin of 5 male and 5 female rats for 24 hours. The rats were observed for mortality, clinical signs of toxicity and behavioral changes daily for 14 days. Body weights were recorded prior to exposure and again on Days 7 and 14. Necropsies were performed on all animals at terminal sacrifice. Under the study conditions, the single dose acute dermal LD50 of HowlerTM Technical was greater than 5,000 mg/kg body weight in male and female rats (Toxicity Category IV).

4. Acute Inhalation Toxicity Study in Rats (OCSPP 870.1300): An acute inhalation study demonstrates that the microbial pest control agent (MPCA), HowlerTM Technical is not toxic by the inhalation route (MRID No. 495680-05). The TGAI, HowlerTM Technical was not toxic to rats following an acute inhalation exposure. An acute inhalation toxicity study was conducted on rats to determine the potential for HowlerTM Technical to produce toxicity from a single 4-hour inhalation exposure. Ten healthy rats (5/sex) were exposed to the test atmosphere in a nose-only chamber for 4 hours. Chamber concentration and particle size distributions of the test atmosphere were determined periodically during the exposure period. The gravimetric chamber concentration was 5.044

mg/L and the average mass median aerodynamic diameter was estimated to be

2.39 μ m. The animals were observed for mortality, clinical signs of toxicity, and behavioral changes daily for 14 days following exposure. Body weights were recorded prior to exposure and again on Days 1, 3, 7, and 14. Necropsies were performed on all animals at terminal sacrifice. Under the study conditions, the single dose acute inhalation LC50 of Howler Technical for a 4-hour exposure was greater than 5.05 mg/L (Toxicity Category IV).

5. Primary Eye Irritation (OCSPP 870.2400): The TGAI, Howler Technical (100% Pseudomonas chlororaphis subsp. aurantiaca strain AFS009), was not an eye irritant following a 24-hour ocular exposure to rabbits. One-tenth of a milliliter of Howler Technical was instilled into the right eyes of three healthy rabbits (MRID No. 495680-06). The left eyes remained untreated and served as controls. Ocular irritation was evaluated using the Draize scoring method. No ocular irritation was observed in any treated eye during the study, and the test substance was classified as non-irritating to the eye (Toxicity Category IV).

6. Primary Dermal Irritation (OCSPP 870.2500): The TGAI, HowlerTM Technical (100% Pseudomonas chlororaphis subsp. aurantiaca strain AFS009),was not a dermal irritant following a 4-hour dermal exposure to rabbits. Fivetenths of a milliliter of Howler Technical was applied to the skin of three healthy rabbits and covered for four hours with a gauze pad and semi-occlusive tape (MRID No. 495680-07). Following exposure, dermal irritation was evaluated using the Draize scoring method. No dermal irritation was observed in any rabbit during the study, and the test substance was classified as non-irritating to the skin (Toxicity Category IV).

7. Hypersensitivity Incidents (OCSPP 885.3400): The registrant has noted that no incidents of hypersensitivity or any other adverse effects have occurred throughthe research, development or testing of the active ingredient and its related enduse product. Should any incidents occur, they will be reported per FIFRA Section 6(a)(2) (MRID No. 495680-16).

Literature searches have demonstrated that there are no reports of ecological or human health hazards caused by Pseudomonas chlororaphis strains. It does not produce recognized toxins, enzymes, or virulence factors normally associated with mammalian invasiveness or toxicity. Pseudomonas chlororaphis is an entomopathogenic fungus and a search of the literature demonstrates it is not reported to be pathogenic to humans. A search of the National Library of Medicine, PubMed, using the terms

“ Pseudomonas chlororaphis” AND “mammal” AND “pathogenicity” resulted in “No items found” (MRID No. 495680-16).

The results of toxicity testing show there is no risk to human health from the active ingredient. Pseudomonas chlororaphis subsp. aurantiaca strain AFS009 is not toxic, pathogenic, infective or irritating to mammals.



### D. Aggregate Exposure

1. Dietary exposure.5



### i. Food. Dietary exposure from use of Pseudomonas chlororaphis subsp.

aurantiaca strain AFS009, as proposed, is minimal. The intended use of Pseudomonas chlororaphis subsp. aurantiaca strain AFS009 is as a biological fungicide to growing plants in greenhouses, agricultural fields and home gardens for the purposes of disease control.

The results of toxicity testing indicate there is no risk to human health or the

environment from Pseudomonas chlororaphis subsp. aurantiaca strain AFS009. There are no reports of ecological or human health hazards caused by this strain

of Pseudomonas chlororaphis . 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 Pseudomonas chlororaphis subsp. aurantiaca strain AFS009 in consumed drinking water would be unlikely. Potential exposure to surface water would be negligible and exposure to drinking water (well or ground water) would be impossible to measure. Pseudomonas chlororaphis is a common bacterium found in soils (MRID No. 495680-16).

The intended use of Pseudomonas chlororaphis subsp. aurantiaca strain AFS009 is to growing plants in greenhouses, agricultural fields and home gardens for the purposes of disease control. The risk of the microorganism passing through the soil to ground water is minimal to unlikely. Additionally the fungus 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 Pseudomonas chlororaphis subsp. aurantiaca strain AFS009. 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 intended use of Pseudomonas chlororaphis subsp.

aurantiaca strain AFS009 is to growing plants in greenhouses, agricultural fields and home gardens for the purposes of disease control. Personal Protective Equipment (PPE) mitigates the potential fo r exposure to applica tors and handlers of the proposed products, when used in agricultural settings. Pseudomonas chlororaphis is a common bacterium found in soils (MRID No. 495680-16).

The results of toxicity testing indicate there is no risk to human health or the

environment from Pseudomonas chlororaphis subsp. aurantiaca strain AFS009. 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. Non-dietary6

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, Pseudomonas chlororaphis subsp. aurantiaca strain AFS009 would result in residues that are of toxicological

concern. The intended use of Pseudomonas chlororaphis subsp. aurantiaca strain AFS009 is to growing plants in greenhouses, agricultural fields and home gardens for the purposes of disease control. The results of toxicity testing indicate there is no risk to

human health or the environment from Pseudomonas chlororaphis subsp. aurantiaca strain AFS009. 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. Acute toxicity studies have shown that Pseudomonas chlororaphis subsp. aurantiaca strain AFS009 is not toxic, pathogenic, infective or irritating to mammals. The intended use of Pseudomonas chlororaphis subsp.

aurantiaca strain AFS009 is to growing plants in greenhouses, agricultural fields and home gardens for the purposes of disease control. The results of toxicity testing indicate

there is no risk to human health or the environment from Pseudomonas chlororaphis subsp. aurantiaca strain AFS009. 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. There is a reasonable certainty of no harm to the general US population from exposure to this active ingredient.



### 2. Infants and children

. As mentioned above, it is not expected that, when used as proposed, Pseudomonas chlororaphis subsp. aurantiaca strain AFS009 would result in residues that are of toxicological concern. There is a reasonable certainty of no harm for infants and children from exposure to Pseudomonas chlororaphis subsp. aurantiaca strain AFS009 from the proposed uses.



### G. Effects on the Immune and Endocrine Systems

To date there is no evidence to suggest that Pseudomonas chlororaphis subsp.

aurantiaca strain AFS009 functions in a manner similar to any known hormone, or that it acts as an endocrine disrupter.



### H. Existing Tolerances

There is no US EPA tolerance or tolerance exemption for Pseudomonas chlororaphis subsp. aurantiaca strain AFS009.7



### I. International Tolerances

A Codex Alimentarium Commission Maximum Residue Level (MRL) is not

established for Pseudomonas chlororaphis subsp. aurantiaca strain AFS009.