Document ID: EPA-HQ-OPP-2014-0919-0002
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2015-01-28T05:00Z

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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, Inc.  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-remove.

Pursuant to section 408(d)(2)(A)(i) of  FFDCA, as amended, Marrone Bio
Innovations, Inc. has submitted the following summary of information,
data, and arguments in support of their pesticide petition. This summary
was prepared by Marrone Bio Innovations, 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. 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 SA-13 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 sequence 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
Muscodor 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 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.

	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 SA-13 are simple alcohols and esters that are
known natural and synthetic flavoring substances. An analytical method
for residues is not required for an exemption from tolerance because
when used as proposed MBI-601 EP would not result in residues of sterile
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 the 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.

Sterile 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 exposure. 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 application 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 Muscodor albus strain SA-13. The volatile compound released from
sterile grain inoculated with Muscodor albus strain SA-13 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 component
of fruits and vegetables.  This flavor is present in apple juice from
three different varieties (Fuji, 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 the current presence of the volatiles in the diets indicate
there is little if any risk to human health from exposure to sterile
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.

ii. 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.

iii. 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 incorporated
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, is 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 concern 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.  Label 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 requires the EPA to consider the
cumulative effects of chemicals with a common mechanism of toxicity in
its tolerance assessment decisions.   SEQ CHAPTER \h \r 1 There is no
indication that sterile grain inoculated with Muscodor albus strain
SA-13, or the volatiles 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   SEQ CHAPTER \h \r 1 toxicity studies have
shown that sterile grain inoculated with 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 following 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.

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