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**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 - 6502 **SUBMISSION: Email**  **the completed template to: hollis**  **. linda**  **@epa.gov. TEMPLATE:**  Marrone Bio Innovations, Inc. 4F8271 EPA has received a pesticide petition 4F8271 from Marrone Bio Innovations, In c. 1540 Drew Avenue , Davis, CA 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 1. by establishing a tolerance for residues of NA - Remove 2. to establish an exemption from the requirement of a tolerance for 3. to establish an amendment/expansion of an existing tolerance exemption for the NA - Remove 1. microbial pesticide sterile grain inoculated with Muscodor albus strain SA - 13 2. biochemical pesticide NA - Remove 3. plant-pesticide NA - remove in or on NA -remov e. Pursuant to section 408(d)(2)(A)(i) of FFDCA, as amended, Marrone Bio Innovations, In c. has submitted the following summary of information, data, and arguments in support of their pesticide petition. This summary was prepared b y Marrone Bio Innovations, Inc. and EPA has not fully

evaluated the merits of the pesticide petition. The summary may have been edited b y 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.2



### I. Marrone Bio Innovations, Inc. Petition Summary 4F8271 A. Product Name and Proposed Use Practices Sterile grain inoculated with Muscodor albus strain SA - 13 is the active ingredient in MBI - 601 EP which is proposed for use as a soil treatment to control or suppress soil -borne plant diseases and plant parasitic nematodes and insects in horticultural and agricultural soils. When activated with moisture, sterile grain inoculated with Muscodor albus strain SA- 13 produces volatile organic compounds that inhibit the growth or kill target pests. MBI - 601 EP is incorporated into the soil before planting or applied in furrow post planting along the planting rows and watered in to activate the volatile compounds. 

### B. Product Identity/Chemistry 1. Identity of the pesticide and corresponding residues . Muscodor albus strain SA13 is a naturally- occurring microbe originally isolated from the honey mesquite host plant in South Africa. Phylogenetic analysis of Muscodor albus strain SA- 13 was performed by acquisition of the ITS - 58S ribosomal gene sequence. The Muscodor albus strain SA - 13 rRNA was compared with rRNAs from other Muscodor species and strains of M. albus . The ITS rRNA se quence had a high similarity with other isolates of M. albus and M. crispans . Muscodor albus strain SA- 13 grows as a white sterile mycelium and does not produce spores or other reproductive structures. The active ingredient , sterile grain inoculated with M uscodor albus strain SA- 13 works to inhibit or kill target pests by production of a number of volatiles. These volatiles are primarily simple alcohols and esters that are known natural and synthetic flavoring substances. Information regarding the name, identity and composition of MBI - 601 EP and the volatiles released upon hydration has been submitted to EPA and can be found in MRID No. 493176 - 01 . 

### 2. Magnitude of residues at the time of harvest and method used to determine the residue. Residues of steri le grain inoculated with Muscodor albus strain SA- 13 are not expected on food or feed items because the active ingredient will not be in direct contact with treated commodities. The seven primary volatiles released by sterile grain inoculated with Muscodor albus strain SA- 13 were identified using GC/MS and are primarily simple alcohols and esters that are known natural and synthetic flavoring substances. When used as proposed, MBI - 601 EP would not result in residues of sterile grain inoculated with Muscodor albus 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 detecting and measuring the levels of sterile grain inoculated with Muscodor albus strain SA- 13 is not needed. Residues of sterile grain inoculated with Muscodor albus strain SA - 13 are not expected on food or feed items because the active ingredient will not be in direct contact with treated commodities. There should be no residues of sterile grain inoculated with Muscodor albus strain SA- 13 because the active ingredient has limited survivability once the nutrient source is exhausted. The primary volatiles produced by sterile grain inoculated with Muscodor albus strain S A- 13 are simple alcohols and esters that are known natural and synthetic flavoring substances. An analytical method for3

residues is not required for an exemption from tolerance because when used as proposed MBI 601 EP would not result in residues of steril e grain inoculated with Muscodor albus that are of toxicological concern. The volatiles compounds that are produced by the active ingredient occur naturally or are known synthetic flavoring substances and dissipate rapidly in soil and water. 

### C. Mammalian Toxicological Profile Acute Oral Toxicity (OCSPP 870.1100): MBI 601 EP was low in toxicity in an acute oral toxicity testing in albino rats following the up - and- down procedure (MRID No. 493176 - 06). Female albino rats received a gavage dose of 5,000 mg/kg of the test article and there were no clinical signs of toxicity. No mortality occurred during the 14 - day observation period. The LD50 for MBI - 601 EP was determined to be >5000 mg/kg body weight, classifying it as a Category IV substance for acute oral toxicity. Acute Dermal Toxicity (OCSPP 870.1200): MBI 601 EP was not toxic in an acute dermal study conducted on albino rats (MRID No. 493176 - 07). Single undiluted doses of 5050 mg/kg body weight of the test article were applied to the intact skin of 10 albino rats and exposure was maintained for 24 hours. There was no mortality observed in the study. All male rats gained weight weekly, as did 2 of the females. Three females either lost or didn’t gain weight during the first or second week of the study . No abnormalities were identified in any of the rats in the gross necropsy at study termination. The LD50 for dermal exposure to MBI - 601 EP is greater than 5,050 mg/kg Acute Inhalation Toxicity (OCSPP 870.1300: A mixture of the seven volatiles released by MBI 601 EP was not toxic in an acute inhalation toxicity study conducted in rats (MRID No. 493176 - 08). The potential for acute inhalation toxicity of the active volatiles released by MBI- 601 EP was evaluated in 5 male and 5 female albino rats. Rats were exposed via inhalation to an atmosphere composed of the primary volatiles released by MBI - 601 in nose - only chambers for four hours. No mortality occurred during or after exposure to a concentration of 5.28 mg/L volatiles. All exposed rats exhibited irregular respiration after the exposure, though they recovered and appeared active and healthy by day 3. All rats gained body weight from day 1 to 14, and no gross abnormalities were identified at the terminal necropsy. The LC50 for inhalation exposure to the active volatiles released by MBI - 601 EP is greater than 5.28 mg/L. Primary Dermal Irritation (OCSPP 870.2500 : The results from an acute dermal irritation study (MRID No. 493176 - 10) demonstrate that MBI - 601 EP is not a dermal irritant. In the acute dermal irritation study, single undiluted doses of 500 mg of MBI - 601 EP were applied to the intact skin of 3 albino rats and moistened with 0.5 ml of water, then covered with a semi permeable dressing for 4 hours. Observations for possible dermal irritation were made at 1, 24, 48 and 72 hours after exposure. Erythema and edema were not reported to occur during the observation period. This study supports an EPA Toxicity Category IV for primary dermal irritation. Dermal Sensitization (OCSPP 870.2600): A skin sensitization study was conducted on 15 male and 15 female albino guinea pigs to determine if MBI - 601 EP elicited a sensitizing reaction (MRID No. 493176 - 11). Naive control group animals remained untreated during the induction phase of the study. Test group animals were treated with 400 mg of test substance moistened with 0.4 mL of water. Test animals were treated once weekly for three weeks, for a total of three treatments. After a two -week rest period, all animals (both groups) were challenged at a virgin test site with an application of 400 mg test substance moistened with 0.4 mL of water. The test substance challenge produced no reaction in test animals or naïve control animals after the4

challenge treatment, and therefore did not elicit a sensitizing reaction in guinea pigs. The MBI 601 EP is not a sensitizer. Acute Pulmonary Toxicity / Pathogenicity (OCSPP 885.3150): MBI - 601 EP was not toxic or pathogenic and showed clearance following pulmonary exposure. An acute study was conducted to evaluate the pulmonary toxicity, pathogenicity and infectivity of MBI - 601 EP following a single pulmonary dose of 0.3 ml / 100 g body weight (approximately 21.45 mg dry mycelial weight (DMW)/rat) (MRID No. 493176 - 05). Animals were observed daily for 21 days after dosing for mortality and signs of toxicity. Tissue and blood samples were collected at interim sacrifices to provide a quantitative record of the clearance pattern of the microbe. No behavioral or physical abnormalities were observed for any of the animals on study, and all necropsy samples were negative for any toxic or pathogenic effects due to exposure. All blood and tissue samples collected were considered clear of any test substance by day 3, given that quantification results on that day yielded two consecutive results below the clearance threshold. MBI - 601 EP was determined to be non-toxic and non-pathogenic to rats following tracheal injection. S terile grain inoculated with Muscodor albus strain SA - 13 produces volatile organic compounds that control or suppress soil -borne plant diseases and plant parasitic nematodes and insects. The volatile compounds occur naturally or are known synthetic flavoring substances. None of these compounds are toxic to humans. 

### D . Aggregate Exposure i. Dietary exposu re. Dietary exposure from use of MBI - 601 EP is expected to be minimal based on the proposed use pattern of incorporation into the soil. The intended use of MBI 601 EP is as a soil pest control agent. The application methods specified on the label for MBI - 601 EP are intended to restrict treatment to target soils. The microbe is firmly attached to the sterile barley substrate that is covered with soil after application and has limited survivability once its carrier nutrient source is exhausted. For this app lication method, the poor survivability of the active ingredient will limit any dietary exposure and precludes dispersion to nontarget areas via drift, run- off or other mechanisms. The results of acute oral, dermal and pulmonary toxicity/pathogenicity testing indicates there is little if any risk to human health or the environment from exposure to sterile grain inoculated with Muscodor albus strain SA- 13. There are no reports of ecological or human health hazards caused by sterile grain inoculated with Musc odor albus strain SA - 13. The volatile compound released from sterile grain inoculated with Muscodor albus strain SA13 in the highest proportion at 30.6% is ethanol which is the alcohol in alcoholic beverages and a solvent exempt from tolerance as an inert ingredient in pesticides. The volatile released in the second highest proportion at 30.1% is 3 -methyl - 1 -butanol, a naturally occurring flavoring com ponent of fruits and vegetables. This flavor is present in apple juice from three different varieties (Fu ji, Gala and Lisgala) at all stages of ripeness in concentrations from 0.66 to 7.27 mg/L (Braga et al., 2013). 3 -methyl - 1 - butanol has also been measured in tomatoes at concentrations from 0.4 to 1.5 mg/L (Ortiz - Serrano & Gil, 2010). The volatile released in the third highest proportion is 2 - methyl - 1 -butanol, which has been measured in young wines at levels of 0.048 to 0.14 mg/L (Wondra & Berovic 2001). In the same wines, 3 -methyl - 1 -butanol levels ranged from 0.126 to 0.482 mg/L. The remaining four volatiles, ethyl acetate, isobutanol, ethyl isobutyrate, and methyl isobutyrate make up 20% of the total released. These include: 1) inert ingredients exempt from tolerance (ethyl acetate, and isobutanol), and 2) FDA approved synthetic flavoring substances (21 CFR 172.515; isobutanol, ethyl isobutyrate and methyl isobutyrate. The results of acute toxicity and pathogenicity testing and the5 current presence of the volatiles in the diets indicate there is little if any risk to human health from exposure to steril e grain inoculated with Muscodor albus strain SA- 13 . The absence of toxicity or pathogenicity in laboratory animals demonstrates the lack of toxicological concern for any incremental increase in dietary exposure from use of MBI 601 EP. 

### i i. Food. Residues of sterile grain inoculated with Muscodor albus strain SA- 13 are not expected on food or feed items because the active ingredient will not be in direct contact with treated commodities. The seven primary volatiles released by sterile grain inoculated with Muscodor albus strain SA - 13 were identified using GC/MS and are primarily simple alcohols and esters that are known natural and synthetic flavoring substances. When used as proposed, MBI - 601 EP would not result in residues of sterile grain inoculated with Muscodor albus that are of toxicological concern. i ii. Drinking water. Similarly , exposure to humans from residues of sterile grain inoculated with Muscodor albus strain SA- 13 consumed in drinking water would be unlikely because the microbe is incorporate d into soil attached to a sterile barley substrate. MBI - 601 EP label instructions require that the product be incorporated into the soil for optimal performance, reducing any likelihood that sterile grain inoculated with Muscodor albus strain SA- 13 would enter surface water (Mileson, 2014; MRID # 493176 - 18 ). Further, sterile grain inoculated with Muscodor albus strain SA- 13 has limited survivability once its carrier nutrient source is exhausted. This short life cycle minimizes the likelihood that sterile grain inoculated with Muscodor albus strain SA - 13 could leach through the soil and enter ground or drinking water in significant iii . Non - dietary exposure . The potential for non- dietary exposure to the general population, including infants and children, i s reasonably low given the primary uses are for agricultural and ornamental treatments, and residential uses are expected to be occasional or periodic. The intended use of MBI - 601 EP is for application to soil to control or suppress soil -borne plant diseases. Sterile grain inoculated with Muscodor albus strain SA- 13 has limited survivability once its carrier nutrient source is exhausted. Volatile compounds released from sterile grain inoculated with Muscodor albus strain SA - 13 are not of toxicological conc ern and dissipate rapidly in the environment. Personal Protective Equipment (PPE) mitigates the potential for exposure to applicators and handlers of the proposed product when used in agricultural settings. L abel mitigation measures are included on packaging for residential use, intended to minimize exposures to infants and children. In addition, the product is incorporated into the soil and not readily available for accidental exposure. E. Cumulative Effects The Food Quality Protection Act of 1996 re quires the EPA to consider the cumulative effects of chemicals with a common mechanism of toxicity in its tolerance assessment decisions. There is no indication that sterile grain inoculated with Muscodor albus strain SA- 13 , or the vo latiles produced by sterile grain inoculated with Muscodor albus strain SA- 13, act by a common mechanism of action with other compounds to result in any cumulative effects F . Safety Determination 1. U.S. population. Acute toxicity studies have shown that sterile grain inoculated with6

Muscodor albus strain SA- 13 and the volatiles released upon rehydration are not toxic or irritating to mammals. These results indicate sterile grain inoculated with Muscodor albus strain SA - 13 is not of toxicological concern for the general population when used as proposed on the label. Further, sterile grain inoculated with Muscodor albus strain SA- 13 was not pathogenic or infective f

ollowing tracheal injection, indicating that sterile grain inoculated with Muscodor albus strain SA- 13 is not of concern for pathogenicity or infectivity when used as proposed. Based on the data available, there is a reasonable certainty of no harm to the general US population from exposure to sterile grain inoculated with Muscodor albus strain SA- 13. 2. Infants and children . As mentioned above, it is expected that, when used as proposed, sterile grain inoculated with Muscodor albus strain SA - 13 would not result in residues that are of concern for potential toxicity or pathogenicity. Based on the data provided, there is a reasonable certainty of no harm for infants and children from exposure sterile grain inoculated with Muscodor albus strain SA- 13 from the proposed uses. 

### G. Effects on the Immune and Endocrine Systems To date there is no evidence to suggest that sterile grain inoculated with Muscodor albus strain SA- 13 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 sterile grain inoculated with Muscodor albus strain SA- 13. 

### I . International Tolerances A Codex Alimentarium Commission Maximum Residue Level (MRL) is not established for sterile grain inoculated with Muscodor albus strain SA - 13.