Document ID: EPA-HQ-OPP-2014-0920-0002
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2015-02-10T05:00Z

EPA BIOPESTICIDES AND POLLUTION PREVENTION DIVISION

NOTICE OF FILING TEMPLATE FOR PESTICIDE PETITIONS PUBLISHED IN THE

FEDERAL REGISTER 

EPA Biopesticides and Pollution Prevention Division contact: Susanne
Cerrelli at (703) 308-8077 

EPA has received a pesticide petition [insert petition number] from
Certis USA LLC, 9145 Guilford Road, Suite 175, Columbia, MD 21046
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 residues
of the microbial pesticide Bacillus mycoides isolate J in or on all
agricultural commodities. 

Pursuant to section 408(d)(2)(A)(i) of the FFDCA, as amended, Certis USA
LLC has submitted the following summary of information, data, and
arguments in support of their pesticide petition. This summary was
prepared by Certis USA LLC 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. Certis USA LLC Petition Summary 

Petition number 4F8252 

A. Product name and Proposed Use Practices 

The microbial pest control agent is Bacillus mycoides isolate J.
Bacillus mycoides isolate J is a naturally occurring, spore-forming
microorganism, one of six species that comprise the Bacillus cereus
group. It was isolated in a field of infected sugar beets. Bacillus
mycoides isolate J is a common, ubiquitous soil bacterium that is found
worldwide.  The mode of action of Bacillus mycoides isolate J is to
induce systemic resistance, triggering the plant’s defenses to control
or suppress plant diseases.

Application of live vegetative cells of Bacillus mycoides isolate J to
the leaves of growing crops induces systemic resistance in the plant
that is characterized by an oxidative burst and production of
pathogenesis related proteins including chitinase and peroxide and beta
glucanase, but does not cause plant death or tissue necrosis.

B. Product Identity/Chemistry 

Identity of the pesticide and corresponding residues. 

The microbial pest control agent Bacillus mycoides isolate J is a
naturally occurring soil organism which will induce systemic resistance
to certain plant diseases. Under proper environmental conditions,
Bacillus mycoides isolate J induces systemic resistance in a broad range
of plants and will be effective in controlling fungal and bacterial
diseases such as sugar beets, cucurbits, tomatoes, ornamentals,
solanaceous plants and other crops.

Bacillus mycoides isolate is a common and ubiquitous soil microorganism
that was first isolated by the Jacobson group from leaves of sugar beets
grown in Montana in 1994.  Bacillus mycoides isolate J forms very
distince spiral colonies on nutrient agar, and is one of six species
that comprise the B.cereus group. A number of DNA sequence matching
techniques have been used to distinguish BmJ from other B.cereus group
isolates; this species is genetically separate from other species in the
group.

The active ingredient can be cultured readily on various culture media
and is cultured by conventional submerged fermentation. Spores of the
fermentation are recovered from the fermentation broth and dried to
create the technical grade active ingredient.  A master stock is
isolated from a single colony isolate by plating on potato dextrose
agar. The culture has been deposited in the Agricultural Research
Service depository in Peoria, IL with the accession number NRRL
B-30890.]

 

Magnitude of residue at the time of harvest and method used to determine
the residue. 

Bacillus mycoides isolate J is a naturally occurring soil microorganism
which has been isolated in agricultural soils worldwide and, as such,
long term exposure to humans, in crops is expected to occur.  No
toxicity or pathogenicity to humans has been reported in the literature
or from tests, resulting from exposure to Bacillus mycoides isolate J,
and this microorganism is not known to be toxic or pathogenic or toxic
to any environmental animal or plant species.  Therefore, when used
according to label directions, Bacillus mycoides isolate J would not
result in residues that are of toxicological concern.  

 

The spores of Bacillus mycoides isolate J are affected by sunlight,
temperature, humidity, substrate, and chemicals.  However, it is the
systemic induction of resistance to certain fungal and bacterial
diseases by the plant that provides control, and not the infectivity of
germinating spores.  The half-life of the spores is relatively short,
and normal washing after harvest and before sale would further reduce or
remove any residues.  

3. A statement of why an analytical method for detecting and measuring
the levels of the pesticide residue are not needed. 

The acute toxicity studies discussed in Section C of this document are
sufficient to show that there are no foreseeable human or domestic
health hazards likely to arise from the use of the product as proposed. 
This soil microorganism is present naturally and Certis USA is
requesting an exemption from the requirement for a tolerance for any
residues remaining in/on all agricultural commodities.  Since
enforcement of residue levels would not be needed, an enforcement
analytical method is not required. 

C. Mammalian Toxicological Profile 

Acute oral toxicity/pathogenicity- rat –To evaluate the acute oral
toxicity, infectivity/pathogenicity of Bacillus mycoides isolate J
Technical Grade Active Ingredient, a single high dose of 6.3 x 109 CFU
per gram or 6.3 x 108 was administered by gavage to rats.  At necropsy,
no internal observable abnormalities were noted, no signs of
toxicological effects.  There was no mortality during the study and the
test material had cleared from brain, mesenteric lymph nodes, blood,
kidney, spleen, liver, lung and caecum 

Acute intravenous toxicity/pathogenicity – rat – the test substance
was administered at a single high dose of 5.3x 108 CFU/gram totwo groups
or rats.  No signs of toxicity or pathogenicity were obse

rved under the condition of the test. No abnormalities were observed for
the duration of the study. The test material cleared completely from the
brain, blood, lungs, kidneys, and cecum.  A pattern of clearance was
established in the liver, 

Acute pulmonary toxicity/pathogenicity- rat –A single intratracheal
dose 1.1 x 108 CFU/ rat was administered to test animals.  No deaths
occurred and there were no signs of toxicity observed in any animal
under the conditions of this test.  Although total clearance of the
microorganism was not established, no clinical signs of toxicity or
infectivity were noted for the duration of the study.

 

Acute oral toxicity –rat- the test substance (BmJ WG) at 5000 mg/kg
was administered via gavage to female rats. No mortality occurred during
the study, and no clinical signs of toxicity were observed.  The acute
LD50 was determined to be greater than 5,000 mg/kg. Based on the test
results, EPA has classified the product at toxicity category IV.

Primary eye irritation – the test substance (BmJ WG) was instilled in
the eye of albino rabbits and examined at 1, 24, 48 and 72 hours after
treatment.  The test substance (produced irritation which was reversible
within 48 hours after treatment.  No irritation was observed in any eyes
at 72 hours. Based on the test results EPA has classified the product as
toxicity category III.

Dermal irritation – rabbit – in a four hour dermal exposure to the
skin of rabbits, the test substance (BmJ WG) was non-irritating with a
primary irritation index of zero.  Based on the test results, EPA has
classified the product as toxicity category IV.

Acute dermal toxicity - rats – the test substance (BmJ WG) was
administered as a single dose at 5050 mg/kg.  No mortality was observed
for the duration of the study, and there were not clinical signs of
toxicity at any time during the study. Based on the test results, the
LD50 for BmJ WG is greater than 5050 mg.kg.

D. Aggregate Exposure 

1. Dietary exposure.   

food

Bacillus mycoides isolate J is a naturally occurring soil microorganism,
which has been isolated in soils worldwide and as such, humans, animals,
and plants are constantly exposed to this microorganism.  It is expected
that, when used as proposed, Bacillus mycoides isolate J would not
result in residues that are of toxicological concern. Bacillus mycoides
isolate is sensitive to environmental factors such as sunlight and
temperature. Bacillus mycoides isolate J spores lose their viability
when exposed to periods of sunlight and temperatures consistent with
mammalian body temperatures. After harvest any remaining spores on crop
leaves will be exposed to unfavorable conditions (e.g. dryness), and are
not likely to germinate; any potentially occurring residual deposits on
these crops will not harm humans because Bacillus mycoides isolate J
demonstrates no toxicity, infectivity, or pathogenicity in appropriate
tests.  Further the optimal temperature range for, germination between
27 and 35 degrees C while the normal body temperatures of humans, other
mammalian species and avian species is significantly higher than 35
degrees C and therefore unsuitable for germination of Bacillus mycoides
isolate J.

While the proposed use pattern may result in dietary exposure with
possible residues in or on certain agricultural commodities, negligible,
to no risk, is expected for the general population, including infants
and children, or animals because Bacillus mycoides isolate J
demonstrated no pathogenicity or toxicity at the maximum doses tested.

Since Bacillus mycoides isolate J is a microorganism that is ubiquitous
in soils, humans may be exposed dermally and orally to this common
fungus when they get soil on their hands or clothing, or handle pets
that have played in soil. Importantly, no toxicological endpoints were
identified for Bacillus mycoides isolate J, and there is no evidence of
adverse effects from oral, dermal, or pulmonary exposure to this MPCA. 
It is not expected that the pesticidal use of Bacillus mycoides isolate
J will significantly increase the non-dietary and non-occupational
exposure to the fungus. 

   

Based on the proposed use patterns, in which Bacillus mycoides isolate J
is applied directly to foliage of agricultural and ornamental crops, the
potential for non-dietary, non-occupational exposures to Bacillus
mycoides isolate J pesticide residues by the general population,
including infants and children, is low. Moreover, even in the unlikely
event of non-dietary, non-occupational exposures to Bacillus mycoides
isolate J residues, no harm is expected because no toxicity or
pathogenicity was found in mammalian studies that included high levels
of oral, pulmonary, and dermal exposure.

. Drinking water. 

Bacillus mycoides isolate J is not known as an aquatic microorganism,
and therefore is no evidence in the literature that this microorganism
will germinate in aquatic habitats. There are no aquatic use sites
permitted for this pesticide, so exposure to drinking water is not
expected. Accordingly, the use of this MCPA on terrestrial plants is not
anticipated to lower the quality of drinking water. Even if low levels
of the microbe were present in drinking water, no risk to the general
public would be expected because Bacillus mycoides isolate J
demonstrated no pathogenicity or toxicity at the maximum dose tested.
The potential of significant transfer to drinking water is minimal to
nonexistent. 

2. Non-dietary exposure.

Since Bacillus mycoides isolate J is a naturally occurring
microorganism, there may be non-dietary exposure, but not above
background levels of this naturally occurring microorganism.   

E. Cumulative Exposure 

There is reasonable certainty that no harm will result to the U.S.
population, including infants and children, from aggregate exposures to
residues of Bacillus mycoides isolate J as a result of its proposed
uses. This includes all anticipated dietary exposures and all other
exposures for which there is reliable information. There appears to be
no potential for harm, from this microorganism in its use as a microbial
pesticide in agricultural commodities. Furthermore, the organism is
naturally occurring in soils, and is non-toxic and non-pathogenic to
animals and humans. This conclusion is based on the results of the
mammalian toxicity tests referenced in Section C. Moreover, potential
non-occupational inhalation or dermal exposure is not expected to pose
any adverse effects to exposed populations via aggregate and cumulative
exposure. 

Therefore no cumulative effects from the residues of this product with
other related microbial pesticides are anticipated. 

F. Determination of Safety for U.S. population, Infants and Children 

There is reasonable certainty that no harm will result to the U.S.
population, including infants and children, from aggregate exposures to
residues of Bacillus mycoides isolate J as a result of its proposed
uses.  This includes all anticipated dietary exposures and all other
exposures for which there is reliable information.  As discussed
previously, there appears to be no potential for harm, from this
microorganism in its use as a systemic inducer of resistance by the
treated plant/crop Furthermore, the organism is naturally occurring in
the environment, and non-toxic and non-pathogenic to animals and humans.
 This conclusion is based on the results of the mammalian toxicity tests
discussed in Section C.  Moreover, potential non-occupational inhalation
or dermal exposure is not expected to pose any adverse effects to
exposed populations via aggregate and cumulative exposure.

G. Effects on the Immune and Endocrine Systems 

There is no information to indicate that Bacillus mycoides isolate J
would have an effect in humans similar to an effect produced by a
naturally-occurring estrogen or other endocrine effects.  There is no
known metabolite produced by this microorganism that acts as an
endocrine disruptor.  The submitted toxicity/pathogenicity studies in
rodents indicate that following dosing no viable spores were found in
organs or tissues of test animals.  It is concluded that there will be
no incremental adverse effects to the endocrine system.

H. Existing Tolerances 

Under 40 CFR 180.1269, there is only a temporary exemption from the
requirement of a tolerance

for Bacillus mycoides isolate J when used as a fungicide on potatoes in
accordance with a valid Federal Insecticide, Fungicide, and Rodenticide
Act (FIFRA) section 18 emergency exemption established. This temporary
exemption from the requirement of a tolerance expires and is revoked on
December 31, 2015.

There is no tolerance or other exemption from the requirement of a
tolerance established for residues of Bacillus mycoides isolate J.

I. International Tolerances 

The European Food Safety Authority Scientific Report (2007), 115, 1-161,
determined that under Regulation (EC) No 396/2005 of the European
Parliament and of the Council on Maximum

Residue Levels for pesticides (28 October 2007) that no MRLs and no
residue definitions have

to be established for the use of Bacillus mycoides isolate J on
agricultural crops.