Document ID: EPA-HQ-OPP-2016-0348-0002
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2016-08-29T04:00Z

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: Kimberly Nesci, 703-308-8095.
Marrone Bio Innovations (EPA Reg. No. 84059)
Petition #6F8444

	EPA has received a pesticide petition #6F8444 from Marrone Bio Innovations, 1540 Drew Avenue, Davis, California 95618 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 the microbial pesticide  Bacillus amyloliquefaciens strain F727.
	Pursuant to section 408(d)(2)(A)(i) of  FFDCA, as amended, Marrone Bio Innovations has submitted the following summary of information, data, and arguments in support of their pesticide petition. This summary was prepared by Marrone Bio Innovations 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. Marrone Bio Innovations Petition Summary
#6F8444
A. Product Name and Proposed Use Practices

Product Names:	MBI-110 TGAI (Technical Grade Active Ingredient; for manufacturing use only)
	MBI-110 EP (End-use Product; for agricultural use, including greenhouse, field and post-harvest uses, for turf use, for home and garden use)
				
Active Ingredient:	Bacillus amyloliquefaciens strain F727 (CAS No. not applicable) 
	Bacillus amyloliquefaciens strain F727 was isolated from a soil sample collected in Northern California. The bacterium was recovered from the sample by traditional plate dilution methods.   
                  Bacillus amyloliquefaciens strain F727 is intended for use in end use products as a fungicide, bactericide and plant health product for agricultural, ornamental, turf, and home and garden uses. 
                  Information regarding the name, identity and composition has been submitted to EPA and can be found in MRID No. 49788501-49788503.
                  The purpose of this petition is to apply for an exemption from the requirement of a tolerance for residues of products containing the active ingredient Bacillus amyloliquefaciens strain F727 in and on all food commodities when used according to good agricultural practices. 
            
Mode of Action:
                  Bacillus amyloliquefaciens strain F727 was isolated from a soil collected in Jonesville, CA using traditional plate dilution methods. Bacillus amyloliquefaciens strain F727 was identified via 16S rRNA amplification and sequencing; amplification and sequencing of housekeeping genes, and characterized using several biochemical assays. This isolate has potential to be used as a biopesticide product for fungicide, bactericide and plant health applications for agricultural, ornamental, turf, and home and garden uses (MRID 49788502-49788503.
                  Bacillus amyloliquefaciens strain F727 has demonstrated activity against bacterial pathogens in vitro and against fungal pathogens both in vitro and in vivo. With in vitro testing, the most inhibited pathogens are Sclerotinia, Macrophomina, Eutypa, Rhizoctonia, and Sclerotium.   
                  Bacillus amyloliquefaciens strain F727 is also a beneficial rhizobacterium that colonizes plant root hairs, leaves and other plant surfaces to prevent establishment of fungal /bacterial plant diseases, and bioassays indicate the bacterium also produces secondary metabolites and/or proteins that inhibit bacterial/mycelial growth and spore germination. It has also been reported that this microbe can induce resistance in plants. 
                  Information on the mode of action has been submitted to EPA and can be found in MRID No. 497885-03
Use Pattern:
            	Bacillus amyloliquefaciens strain F727 is intended as a foliar spray,  soil treatment or seed treatment to control fungal and bacterial diseases and to promote plant health on numerous agricultural crops, nursery crops, ornamental plants and turfgrass in both field and greenhouse conditions.  
            
B. Product Identity/Chemistry
 		Identity of the Pesticide and Corresponding Residues  
                    Not applicable.
 		Magnitude of Residues at the Time of Harvest and 				Method Used to Determine the Residue
       		Not applicable.
 		Statement of Why an Analytical Method of Detecting and 			Measuring the Levels of the Pesticide Residue are not 			Needed
                  An analytical method for detecting and measuring the levels of the pesticide residue for Bacillus amyloliquefaciens strain F727 is not applicable.  It is expected that, when used as proposed, Bacillus amyloliquefaciens strain F727 would not result in residues that are of toxicological concern. 

C. Mammalian Toxicological Profile

Mammalian Toxicological Profile

            A variety of toxicity studies have been performed with Bacillus amyloliquefaciens strain F727.  The toxicity studies performed are listed below in Table 1 and include an acute oral toxicity study in rats, an acute dermal toxicity study in rats, an acute inhalation toxicity study in rats, an acute intravenous toxicity study in rats, an acute dermal irritation study in rabbits, and an acute eye irritation study in rabbits. The oral, dermal and inhalation routes of exposure are the routes relevant for human health risk assessment. When mammals were exposed through oral and dermal routes, the results of these studies consistently showed that Bacillus amyloliquefaciens strain F727 was non-toxic and also lacked irritation to eyes and skin.  Following inhalation of undiluted Bacillus amyloliquefaciens strain F727, the test substance was rated as non-toxic in rats. Given that human inhalation exposure would not be to undiluted liquid substance, the relevance of these data to risks associated with undiluted Bacillus amyloliquefaciens strain F727 is minimal. The results of the intravenous toxicity study showed that Bacillus amyloliquefaciens strain F727 containing Bacillus amyloliquefaciens strain F727 cells was not pathogenic in mammals. No reports of hypersensitivity have been recorded from personnel working with this organism.  As evidenced by the results of toxicity testing summarized below in Table 1, there is no anticipated risk to human health following exposure to Bacillus amyloliquefaciens strain F727 through oral, dermal or inhalation routes.   
TABLE 1      Summary of Bacillus amyloliquefaciens strain F727 Mammalian Toxicity Data 
MRID
Mammalian Toxicity Test
Species
Dose Tested
Results   

497885-06
Acute oral toxicity

Rat
5000 mg/kg

LD50 >5000 mg/kg
Category IV

497885-07
Acute dermal toxicity

Rat

5050 mg/kg

LD50 >5050 mg/kg
Category IV

497885-08
Acute inhalation toxicity

Rat
5.19 mg/L (undiluted liquid test substance for 4 hours)
LC50 = >5.19mg/L
Category IV
497885-04
Acute intravenous pathogenicity-toxicity
Rat

2.9 x 109 cfu/mL

Non-toxic, non-pathogenic; Clearance in blood by  Day 3; clearance in brain and cecum contents by Day 7; and clearance in lungs, spleen, kidneys and lymph nodes by Day 14 

497885-10
Acute dermal irritation

Rabbit

0.5 mL undiluted product for 4 hours
Non-irritating
Category IV

497885-09
Acute eye irritation

Rabbit

0.1 mL undiluted product for 24 hours
Minimally-irritating
Category IV

Literature searches have demonstrated that there are no reports of ecological or human health hazards caused by Bacillus amyloliquefaciens strains.  It does not produce recognized toxins, enzymes, or virulence factors normally associated with mammalian invasiveness or toxicity.   The results of toxicity testing show there is no risk to human health from the active ingredient.  Bacillus amyloliquefaciens strain F727 is not toxic, pathogenic, infective or irritating to mammals.

D. Aggregate Exposure

      1)  Dietary Exposure

            Dietary exposure from use of Bacillus amyloliquefaciens strain F727, as proposed, is minimal based on the proposed pre-harvest use pattern. The results of toxicity testing following acute oral exposure, the route most relevant to assessment of risks to health from exposure to food, indicate there is no risk to human health from Bacillus amyloliquefaciens strain F727. There are no reports of human health hazards related to oral exposure to Bacillus amyloliquefaciens strain F727 cells. The absence of acute toxicity in laboratory animals following oral exposure to high doses of the product demonstrates the benign nature of Bacillus amyloliquefaciens strain F727.
	Bacillus species, including B. amyloliquefaciens, are commonly found in soils, including in agricultural settings, and are naturally present on fresh produce.  The Manual of Clinical Microbiology (9[th] Edition) states that dried foods, such as spices, milk powder and grains, often contain large amounts of Bacillus spores.  B. amyloliquefaciens is not known to produce any mammalian toxins, and no food-borne disease outbreaks association with B. amyloliquefaciens have been reported.  B. amyloliquefaciens is not recognized as a pathogen among Bacillus species (Logan and de Vos, 2009).
            Based on the data and other information submitted to satisfy the data requirements for registration of the manufacturing-use and end-use pesticide products containing the active ingredient, B. amyloliquefaciens strain F727, no toxicity, infectivity, pathogenicity or other adverse effects from dietary exposure are expected.

      2)  Drinking Water Exposure

            Similarly, exposure of humans to residues of Bacillus amyloliquefaciens strain F727 in drinking water would be minimal based on the proposed application rates of a maximum of 4 quarts of formulated product per acre applied by broadcast, in-furrow, banded, injected, drench or drip irrigation. Results of the acute oral toxicity testing described above have shown that Bacillus amyloliquefaciens strain F727 was non-toxic at the highest test dose (> 5000 mg/kg).   There are no reports of human health hazards related to oral exposure to Bacillus amyloliquefaciens strain F727 cells. The absence of acute toxicity in laboratory animals following oral exposure to high doses of the product demonstrates the benign nature of Bacillus amyloliquefaciens strain F727.
            Bacillus species, including B. amyloliquefaciens, are commonly found in soils, including in agricultural settings, and are naturally present on fresh produce (Logan and de Vos, 2009).  The Manual of Clinical Microbiology (9[th] Edition) states that dried foods, such as spices, milk powder and grains, often contain large amounts of Bacillus spores. According to the World Health Organization, Bacillus species are often detected in drinking water even after going through acceptable water treatment processes, largely because the spores are resistant to drinking water disinfection processes (World Health Organization, 2011) B. amyloliquefaciens is not known to produce any mammalian toxins, and no food-borne disease outbreaks association with B. amyloliquefaciens have been reported.  Should Bacillus amyloliquefaciens strain F727 be present, no adverse effects are expected from exposure to B. amyloliquefaciens strain F727 through drinking water based on the data and other information submitted to satisfy the data requirements for registration of the manufacturing-use and end-use pesticide products containing active ingredient, B. amyloliquefaciens strain F727.

 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 application to growing plants or crops.
            
            The results of toxicity testing discussed above indicate there is no risk to human health or the environment from Bacillus amyloliquefaciens strain F727. There are no reports of ecological or human health hazards related to oral exposure to Bacillus amyloliquefaciens strain F727 cells. The absence of acute toxicity in laboratory animals following oral or dermal exposure to high doses of Bacillus amyloliquefaciens strain F727 demonstrates the benign nature of the product. As a result, non-dietary exposures would not be expected to pose any quantifiable risk.

E. Cumulative Effects

 Cumulative Effects

            It is not expected that, when used as proposed, Bacillus amyloliquefaciens strain F727 would result in residues that are of toxicological concern. The intended use of the product is to apply Bacillus amyloliquefaciens strain F727 as a plant disease control for food and non-food crops. The results of toxicity testing indicate there is no risk to human health or the environment from Bacillus amyloliquefaciens strain F727. There are no reports of ecological or human health hazards related to exposure to Bacillus amyloliquefaciens strain F727 cells. The absence of acute toxicity in laboratory animals following exposure to high doses of the product demonstrates the benign nature of Bacillus amyloliquefaciens strain F727. The lack of acute toxicity indicates that the hazard associated with the use of Bacillus amyloliquefaciens strain F727 is low.

F. Safety Determination

 U.S. Population 
         
            Acute toxicity studies have shown that the Bacillus amyloliquefaciens strain F727 is non-toxic in mammals when exposure is through oral and dermal routes and also lacks irritation potential in mammals.  Further, testing in mammals of active microbes showed that the product was not pathogenic. There are no reports of ecological or human health hazards caused by Bacillus amyloliquefaciens strain F727 cells. The lack of acute toxicity indicates that the hazard associated with the use of Bacillus amyloliquefaciens strain F727 is low.  There is a reasonable certainty of no harm to the general United States population from exposure to this active ingredient.
            
            
            
 Infants and Children
            	
            As mentioned above, it is not expected that, when used as proposed, Bacillus amyloliquefaciens strain F727 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 amyloliquefaciens strain F727 from the proposed uses.

G. Effects on the Immune and Endocrine Systems
      
            To date, there is no evidence to suggest that Bacillus amyloliquefaciens strain F727, functions in a manner similar to any known hormone, or can act as an endocrine disrupter.

H. Existing Tolerances
            There are no current tolerances or tolerance exemptions in place for Bacillus amyloliquefaciens strain F727.

I. International Tolerances
 
            A Codex Alimentarium Commission Maximum Residue Level (MRL) is not established  for Bacillus amyloliquefaciens strain F727.