Document ID: EPA-HQ-OPP-2017-0294-0002
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2017-11-06T05:00Z

<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: [Michael Glikes, 703-305-6231]>

<SUBMISSION: E-mail the completed template to: carlisle.sharon@epa.gov>

<TEMPLATE:>

<International Animal Health Products Pty. Ltd.>

<[  ]>

<	EPA has received a pesticide petition from International Animal Health Products Pty. Ltd., 18 Healey Circuit, Huntingwood, NSW 2148, Austalia, 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 >

<(Options (pick one)>
	to establish an exemption from the requirement of a tolerance for 

<(Options (pick one)>
   
   	1. microbial pesticide Duddingtonia flagrans strain IAH 1297

<	Pursuant to section 408(d)(2)(A)(i) of FFDCA, as amended, International Animal Health Products Pty. Ltd. has submitted the following summary of information, data, and arguments in support of their pesticide petition. This summary was prepared by SciReg, 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. International Animal Health Products Pty. Ltd. Petition Summary>
   	
<A. Product Name and Proposed Use Practices>

<	BioWorma is a feed supplement containing 5 x 10[5] viable spores/gram of Duddingtonia flagrans strain IAH 1297.  
	Livamol with BioWorma is a feed supplement containing 3 x 10[4] viable spores/gram of Duddingtonia flagrans strain IAH 1297.
	
<B. Product Identity/Chemistry>

<	1. Identity of the pesticide and corresponding residues. Duddingtonia flagrans strain IAH 1297 is a natural ubiquitous soil fungus that was isolated from the environment as part of a field survey of nematophagous fungi in Australia. 

The taxonomic grouping is as follows:
Class: Orbiliomycetes
Order: Orbiliales
Family: Orbiliaceae

Duddingtonia flagrans strain IAH 1297 is identified by its DNA sequence.

	Duddingtonia flagrans strain IAH 1297 is a naturally-occurring fungus that is commonly found in soil and on pasture. It is a non-chemical biological control for the free-living stages of parasitic gastrointestinal nematodes of grazing animals. As animals consume D. flagrans, the thick-walled fungal spores remain intact through digestion and pass through the animal's intestinal tract into the manure. There they germinate and form hyphal structures that trap, paralyze, and consume emerging nematode larvae.

<	2. Magnitude of residues at the time of harvest and method used to determine the residue. No magnitude of residue (MOR) studies have been conducted with Duddingtonia flagrans strain IAH 1297. Based on the low toxicity of the active ingredient strains (see Toxicology Profile, pages 3-6). OPPTS Guideline 885.2000, section f, Background for Residue Analysis of Microbial Pest Control Agents (MCPA), states: 

"The Agency evaluates residue data for MCPAs used on food, feed, or raw agricultural commodities only if toxic or other harmful properties were observed in the maximum hazard toxicology tests (Tier I) prescribed in toxicology test guidelines OPPTS 885.3050 through 885.3650 of this series. If Tier I toxicology tests indicate no toxic or other harmful properties, then no residue data (with the general exception of a monitoring method) would be indicated and thus a recommendation for an exemption from the requirements of a tolerance can be made.">

<	3. A statement of why an analytical method of detecting and measuring the levels of the pesticide residue are not needed. SciReg, Inc. is submitting a petition to establish an exemption from the requirement of a tolerance and, therefore, an analytical method is not required.>

<C. Mammalian Toxicological Profile>

<	The full reports and waiver requests that support this petition and which are summarized in this section were submitted to EPA in support of registration of two end-use products, BioWorma and Livamol with BioWorma.

Acute Oral Toxicity: Three female CRL:(WI) rats were dosed per os with Duddingtonia flagrans strain IAH 1297 at a  rate of 5000 mg/kg. The test substance had a purity of 3.8 x 10[6] spores/gram. No clinical signs were observed and there were no treatment related effects on body weight or body weight gain. No macroscopic observations were noted during necropsy. The oral LD50 of D. flagrans strain IAH 1297 in rats is considered to be great than 5000 mg/kg.

Acute Pulmonary Toxicity/Pathogenicity: Prior to the commencement of an acute pulmonary toxicity and infectivity study, a validation test was carried out with Duddingtonia flagrans dry spore powder on rats. The average instilled quantity of test item was 0.013 g/instillation step and the average number of spores in the test item was 3.8x10[6] spores/gram. The animals were unable to tolerate more than one instillation of both activated and attenuated spores due to the size of the spores physically obstructing airways within a few hours of administration; therefore, it was not possible to achieve the guideline-recommended dose of 1x10[8] cfu/animal. It was concluded that for the main study, animals could be treated with a maximum dose of 0.017 g test item/animal as a single instillation.

An acute pulmonary toxicity and infectivity study was conducted with CRL:(WI) rats. Duddingtonia flagrans strain IAH 1297 was administered to thirty male and thirty female rats at an average quantity of 0.018 grams of test item per animal, or 5.8x104 spores per animal. The maximum treatment dose was determined in a preliminary validation study. During the validation study, two deaths occurred. Test substance instillation was performed in one step. 

Animals treated with both living and attenuated spores exhibited increased respiration rate and labored and noisy respiration. After treatment, these symptoms gradually reduced. Clinical observations also included decreased activity, piloerection, red discharge, and hunched back.

After an initial decrease in body weights, animals returned to their original body weights by Day 7. No treatment-related effects on body weights or body weight gain were noted after one week.

Animals sacrificed on Day 3, 21, or 42 had enlarged lungs, enlarged peribronchial lymph nodes, and multifocal dark red discoloration. Yellow-green mucoid material was noted on the cut surface of the lungs in one animal necropsied on Day 21 that received attenuated spores and one rat necropsied on Day 42 treated with living spores. These findings are not considered to be of toxicological significance.

Spores of Duddingtonia flagrans strain IAH 1297, instilled into the broncho-alveolar space of the experimental animals, caused diffuse purulent pneumonia and multifocal lesions with necrotic area of the pulmonary tissue. Duddingtonia flagrans spores were found in the trachea and lungs. No morphological differences were observed between the changes caused by active or inactivated spores. Further, there was no evidence of reproduction or proliferation of Duddingtonia spores in the pulmonary tissue. Necropsy observations confirmed that spore deposits spread within the lungs following insufflation.

Complete clearance of living spores from the lungs was confirmed at Day 42 when the cfu/g values were zero. The physical properties, such as the large size of the spores (~20 μm), likely resulted in the slow clearance from the lungs.

Blood: All samples negative for Duddingtonia flagrans strain IAH 1297
Brain: All samples negative for Duddingtonia flagrans strain IAH 1297
Lung: Day 42 samples negative for Duddingtonia flagrans strain IAH 1297
Liver: All samples negative for Duddingtonia flagrans strain IAH 1297
Kidney:  All samples negative for Duddingtonia flagrans strain IAH 1297
Spleen: All samples negative for Duddingtonia flagrans strain IAH 1297
Lymph nodes: All samples negative for Duddingtonia flagrans strain IAH 1297
Cecum: All samples negative for Duddingtonia flagrans strain IAH 1297

Based on the results of this study, spores of Duddingtonia flagrans strain IAH 1297 do not induce signs of toxicity, infectivity, or pathogenicity when administered to rats as a single intratracheal dose at the maximum attainable dose (5.8x104 spores per animal). The size and physical surface of the spores blocking the airways, rather than their virulence, determine the maximum dose. This is further confirmed by the body weights increasing at the cessation of clinical signs in animals treated with either living or attenuated spores. It is also noted, that after 42 days, living spores could not be detected in the lungs of the treated animals.

Acute Injection Toxicity/Pathogenicity: The physical form of D. flagrans prohibits the injection of intact spores due to the large spore size (approximately 20 microns). In addition, infectivity in warm-blooded animals is inhibited as D. flagrans cannot germinate at 37 °C or greater or under anaerobic conditions. Further, the intratracheal instillation study of D. flagrans strain IAH 1297 found no signs of toxicity, infectivity or pathogenicity. Infectivity in mammals and birds has never been reported, in spite of the wide-spread proliferation of D. flagrans. Extensive history of human use during the course of this product's development has not shown any evidence of infection among handlers or users of the TGAI or end-use products in Australia and New Zealand. Based on this review of the history of D. flagrans, infectivity via injection is considered improbable.

In addition, D. flagrans strain IAH 1297 is a naturally occurring fungus that is commonly found in soil and on pasture. D. flagrans strain IAH 1297 is not known to germinate or grow at 37o C or greater or under anaerobic conditions.

Acute Dermal Toxicity: Five male and five female CRL:(WI) rats were treated with a single semi-occlusive dermal application of Livamol with BioWorma at 5000 mg/kg. The test substance was applied to a shaved area of the back of the animal for 24 hours, followed by a 14-day observation period. No adverse clinical signs were noted in any animals and no treatment-related skin irritation was observed. There were no deaths and no internal or external observations noted during necropsy. The dermal LD50 of Duddingtonia flagrans strain IAH 1297 in rats is considered to be greater than 5000 mg/kg.

Eye Irritation: Livamol with BioWorma was administered to three male New Zealand White rabbits to investigate acute eye irritation of the test substance. Test animals received 0.1 g of Livamol with BioWorma into the conjunctival sac of the left eye. Animals were observed 1, 24, 48 and 72 hours after test material installation. Chemosis was noted in one animal at 24 hours and conjunctival redness was observed in all animals up to 48 hours. No clinical signs and no conjunctival or corneal effects were observed after 72 hours. Livamol with BioWorma is classified as Toxicity Category III for eye irritation. 

Dermal Irritation: An acute dermal irritation study was conducted using 3 male New Zealand White rabbits. Animals received a topical, semi-occulsive application of 0.5 g of Livamol with BioWorma to their intact shaved flanks. The test substance was administered for 4 hours. Skin reactions were scored at 1, 24, 48, and 72 hours following removal of the dressing. 

No mortality or adverse clinical signs were observed after treatment with the test item or during the 14-day observation period; no effects were observed at the site of application. Body weights were within the range commonly recorded for this strain and age and there were no treatment related body weight changes. No evidence of test item-related observations were noted at necropsy. Livamol with BioWorma is considered Toxicity Category IV for dermal irritation at  a dose level of 5000 mg/kg. 

Hypersensitivity Incidents: No reports of hypersensitivity have been associated with of Duddingtonia flagrans strain IAH 1297.>

<D. Aggregate Exposure>

<	1. Dietary exposure. The potential for non-occupational, dietary, and non-dietary exposure Duddingtonia flagrans strain IAH 1297 by the general population, including infants and children, is possible through exposure to pastures where treated animals defecate.>

<	i. Food. Dietary exposure to Duddingtonia flagrans strain IAH 1297 should not be of concern due to the low toxicity shown across the entire toxicological profile (see above). If Duddingtonia flagrans strain IAH 1297 is ingested, none of the studies have indicated any pathogenicity of the organism in mammals. In addition, the subject strain does not replicate in mammalian cells.>

<	ii. Drinking water. Duddingtonia flagrans strain IAH 1297 does not pose a risk to drinking water. Anticipated agricultural practice prohibits direct application of the end-use product directly to water. Duddingtonia flagrans strain IAH 1297 is not likely to be present in aquatic environments since it is not known to inhabit aquatic environments. Duddingtonia flagrans strain IAH 1297 is generally not present in underground water sources.>

<	2. Non-dietary exposure. Non-dietary exposure to Duddingtonia flagrans strain IAH 1297 will be much more likely through handling by formulators and applicators; however, in light of the subject strain's toxicological profile, should be of little concern. 

<E. Cumulative Effects>

<	Section 408(b)(2) of the FFDCA requires that, when considering whether to establish an exemption from a tolerance, the Agency considers available information concerning the cumulative effects of the particular ingredient's residues and other substances that have a common mechanism of toxicity. These considerations include the possible cumulative effects of such residues on infants and children. 

Duddingtonia flagrans strain IAH 1297 is effective by trapping nematodes with specialized adhesive hyphal networks. The chlamydospores of D. flagrans are incorporated into the livestock feed. Thick walls protect ingested spores, which survive passage through the animals' digestive tract, are deposited through feces into pastures. In pasture, the spores germinate and capture, paralyze, and consume emerging worm larvae including Haemonchus placei, H. contortus Ostertagia ostertagi, Trichostrongylus spp., and Cooperia spp. It is likely grazing animals are naturally exposed to the fungus as it is considered ubiquitous in the environment.>

<F. Safety Determination>

<	1. U.S. population. Duddingtonia flagrans strain IAH 1297 exhibits low mammalian toxicity as demonstrated by the toxicological profile. When Duddingtonia flagrans strain IAH 1297 is incorporated into BioWorma or Livamol with BioWorma in accordance with good agricultural practice, it meets the reasonable certainty of no harm requirement. 

<	2. Infants and children. It is believed that the potential exposure of infants and children to residues of Duddingtonia flagrans strain IAH 1297 through its use to protect grazing animals against parasitic nematodes is no greater than that of the general population. In addition, there is no information or data to suggest that infants and children will be disproportionately at risk to the use of BioWorma or Livamol with BioWorma. Available information does not suggest that infants and children are more susceptible to the effects of Duddingtonia flagrans strain IAH 1297 than are adults.

<G. Effects on the Immune and Endocrine Systems>

<	Extensive literature searches and review of Duddingtonia flagrans strain IAH 1297 toxicity data indicate there is no evidence that it is an endocrine disruptors or that it has any known effect on immune systems.>

<H. Existing Tolerances>

<	There are no known U.S. tolerances or tolerance exemptions existing for Duddingtonia flagrans strain IAH 1297.>

<I. International Tolerances>

<	There are no known international tolerances or tolerance exemptions existing for Duddingtonia flagrans strain IAH 1297.

         
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