Document ID: EPA-HQ-OPP-2010-0104-0002
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2010-03-11T05:00Z

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: [Denise
Greenway, (703) 308-8263] 

EPA has received a pesticide petition [9F7643] 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 subtilis strain CX-9060 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 

	Pesticide Petition Number:  9F7643

A. Product name and Proposed Use Practices 

Bacillus subtilis CX-9060 microbial pest control product is a
broad-spectrum preventative biofungicide for control or suppression of
fungal and bacterial plant diseases.  The product contains a naturally
occurring strain  of the beneficial rhizobacterium Bacillus subtilis,
which colonizes leaves, roots, and other plant surfaces, killing
pathogenic organisms by means of antibiotic compounds (iturins) which
disrupt pathogen cell wall production.  Bacillus subtilis rapidly
colonizes plant root hairs, leaves, and other surfaces, preventing
establishment of disease-causing fungi and bacteria.

The proposed uses for Bacillus subtilis CX-9060 include application
alone or in combination and/or rotation with chemical fungicides as a
tool for integrated disease management in agricultural crops, ornamental
and nursery plants, and turfgrass, (pre- and post-harvest applications).

 Bacillus subtilis CX-9060 offers a valuable tool for management of
resistance to chemical fungicides through its multiple and unique modes
of action.

B. Product Identity/Chemistry 

Identity of the pesticide and corresponding residues. 

Bacillus subtilis (spp.) is a rod shaped,   HYPERLINK
"http://en.wikipedia.org/wiki/Gram-positive" \o "Gram-positive" 
Gram-positive ,   HYPERLINK "http://en.wikipedia.org/wiki/Catalase" \o
"Catalase"  catalase -positive   HYPERLINK
"http://en.wikipedia.org/wiki/Bacterium" \o "Bacterium"  bacterium 
commonly found in soil.  Bacillus subtilis has the ability to form a
tough, protective   HYPERLINK "http://en.wikipedia.org/wiki/Endospore"
\o "Endospore"  endospore , allowing the organism to tolerate extreme
environmental conditions. 

Bacillus subtilis MPCA is manufactured by growing Bacillus subtilis
strain CX9060.  This strain was isolated from existing cultures of
Bacillus subtilis.  The culture was fermented under aeration in liquid
culture medium until sporulation is completed. The culture is deposited
at USDA, ARS-NCAUR, 1815 North University Street, Peoria, Illinois 61604
USA.  The accession number is NRRL-B-50287.

Bacillus subtilis spp. are not considered to be a human pathogen. 
Bacillus subtilis produces proteolytic enzymes and is widely used as an
additive in   HYPERLINK "http://en.wikipedia.org/wiki/Laundry_detergent"
\o "Laundry detergent"  laundry detergents .  Bacillus subtilis is used
as a   HYPERLINK "http://en.wikipedia.org/wiki/Soil_inoculant" \o "Soil
inoculant"  soil inoculant  in   HYPERLINK
"http://en.wikipedia.org/wiki/Horticulture" \o "Horticulture" 
horticulture  and   HYPERLINK "http://en.wikipedia.org/wiki/Agriculture"
\o "Agriculture"  agriculture .  

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

Bacillus subtilis is ubiquitous in the environment, especially in soils
and agricultural environments. As a result, dietary exposure to
background levels of the naturally occurring microbe likely is already
occurring and likely will continue to occur. Because of its ubiquitous
presence in the environment, no increase in exposure to Bacillus
subtilis resulting from the existing and proposed pesticidal uses when
compared to existing exposure to background levels of Bacillus subtilis
is expected to occur. 

Any potentially occurring residual deposits on crops will not harm
humans because Bacillus subtilis strains show no toxicity, infectivity,
or pathogenicity in appropriate tests.  

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. 
Certis USA is requesting an exemption from the requirement for a
tolerance for any residues remaining in/on all agricultural commodities
from pre-and post-harvest applications..  Since enforcement of residue
levels would not be needed, an enforcement analytical method is not
required.

C. Mammalian Toxicological Profile 

Toxicological data on multiple strains of Bacillus subtilis were
submitted to EPA to support an existing exemption from the requirement
of a tolerance for residues resulting from its use in or on all
agricultural crops. New studies on formulated Bacillus subtilis CX-9060,
and previously submitted Bacillus subtilis data, include the following: 

An acceptable acute oral toxicity/ pathogenicity study performed in rats
demonstrated the lack of mammalian toxicity at high levels of exposure
to Bacillus subtilis.  In this study, Bacillus subtilis was not toxic,
infective nor pathogenic to rats given an oral dose of 2 x 108 colony
forming units (CFU) per animal. The study resulted in a classification
as Toxicity Category IV. 

An acceptable acute pulmonary toxicity/pathogenicity study in rats
demonstrated that Bacillus subtilis was neither toxic, pathogenic nor
infective to rats dosed intratracheally with 3.4 x 108 CFU of the test
material. The study resulted in a classification of Toxicity Category
IV. 

An acceptable acute intravenous injection toxicity/pathogenicity study
in rats demonstrated that Bacillus subtilis was neither toxic,
pathogenic nor infective to rats dosed intravenously with approximately
4 x 107 CFU of the test material. Although the microbe was detected in
every organ tested, the test material displayed a distinct pattern of
clearance. The study resulted in a classification of Toxicity Category
IV. 

A new acute dermal irritation study in rabbits demonstrated that
formulated Bacillus subtilis CX-9090 was not toxic to rabbits when a
single 500 mg dose was administered dermally.  Based on the scores at
the 72-hour observation, the study resulted in a classification of
Toxicity Category IV. 

In addition a new primary eye irritation study in rabbits demonstrated
that formulated Bacillus subtilis CX-9090 produced a slight ocular
irritation when a single 0.1 mL by volume of the undiluted test
substance was administered. Since all positive ocular effects cleared by
48 hours, the test substance resulted in a classification of Toxicity
Category III. 

A previously submitted, acceptable primary eye irritation study
demonstrated that Bacillus subtilis produced a slight ocular irritation
when a single 0.1 gram ocular dose was administered.  Ocular irritation
dissipated by day 4.  The study resulted in a classification of Toxicity
Category IV.

A previously submitted, acceptable acute dermal toxicity study in
rabbits, demonstrated that Bacillus subtilis was not toxic to rabbits
when a single 5 x1010 dose was administered dermally.  The study
resulted in a classification of Toxicity Category IV.

A previously submitted supplemental skin sensitization test resulted in
an overall moderate reaction in guinea pigs 24 to 78 hours
post-treatment. However, an acceptable dermal sensitization study,
conducted with an end use formulation, demonstrated no irritation 2
weeks after sensitization and treatment using 400 milligrams of test
material. As a result, the product was determined to not be a dermal
sensitizer. Furthermore, in the nearly 15 years since its initial
registration as an active ingredient, there has been no hypersensitivity
reports associated with Bacillus subtilis pesticide products.

D. Aggregate Exposure 

1. Dietary exposure.   

food

Bacillus subtilis is ubiquitous in the environment, especially in soils
and agricultural environments. As a result, dietary exposure to
background levels of the naturally occurring microbe likely is already
occurring and likely will continue to occur. Because of its ubiquitous
presence in the environment, no increase in exposure to Bacillus
subtilis resulting from the existing and proposed pesticidal uses is
expected when compared to existing exposure to background levels of
Bacillus subtilis. 

As discussed above, dietary exposure to the naturally occurring microbe
likely is already occurring and likely will continue to occur. Notably,
similar Bacillus subtilis strains are used internationally in the
production of food grade products and in fermented foods in Japan and
Thailand. Reports in the literature implicating Bacillus subtilis in
food-borne illness do not describe any pathogen or toxin production, but
rather simple spoilage from Bacillus subtilis growth in dough. Such
low-quality dough would not be suitable for bread production by
commercial bakeries and so Certis USA considers this particular food
exposure scenario to be unlikely and the risk to be negligible. The risk
posed to adults, infants and children from food-related exposures to
Bacillus subtilis is minimal due to the demonstrated lack of acute oral
toxicity/pathogenicity associated with the microbial pesticide. Based on
the submitted data, Certis USA concludes that there are no dietary risks
that exceed the Agency’s level of concern. 

  

. Drinking water. 

Because Bacillus subtilis is ubiquitous in the environment, exposure to
the microbe through drinking water may already be occurring and likely
will continue to occur. While the proposed use sites do not include
direct application to aquatic environments, the intended use of Bacillus
subtilis CX-9060 is treatment of growing crops, pre- and post-harvest
applications, and/or seed for the control of plant disease. If such uses
were to result in pesticide spray drift or runoff that were to reach
surface or ground waters, there is the potential for human exposure to
Bacillus subtilis residues, albeit greatly diluted, in drinking water.
Municipal drinking water treatment processes and deep water wells,
however, would both further reduce any such residues. More importantly,
even if oral exposure to this ubiquitous microbe should occur through
drinking water, due to its demonstrated lack of acute oral toxicity/
pathogenicity, Certis USA concludes that there is a reasonable certainty
that no harm will result from such exposure. 

2. Non-dietary exposure.

The pesticide uses of Bacillus subtilis  both those currently allowed
and the additional proposed uses are limited to commercial agricultural
and horticultural settings. There are no residential uses. Nonetheless,
because Bacillus subtilisis naturally occurring and ubiquitous in the
environment, the potential for non-dietary, non-occupational exposure to
its residues for the general population, including infants and children,
is likely since populations have probably been previously exposed (and
likely will continue to be exposed) to background levels of the microbe.
However, neither such common human exposures to Bacillus subtilis
naturally present in soils, waters and plants, nor exposures associated
with similar Bacillus subtilis strains used internationally in producing
food-grade products and fermented foods, have resulted in reports of
disease or other effects. Finally, while the literature includes
accounts of Bacillus subtilis infections in humans (which consistently
are reported only in otherwise-compromised individuals), those reports
are most notable for their rare and exceptional nature. 

EPA’s previous evaluation of the high-dose Tier I acute toxicity and
pathogenicity tests resulted in the assignment of Toxicity Category IV
(least toxic), and determinations of not infective and not pathogenic,
for all exposure routes. No toxicological end points of concern were
identified. There are no dietary endpoints that exceed EPA’s Level of
Concern (LOC). Certis USA notes that the Agency has previously
determined that any additional exposure to the microbe resulting from
residues attributable to Bacillus subtilis pesticide use will not result
in additional aggregate non-occupational risk from dermal and inhalation
exposures. This conclusion, based solely on non-occupational exposures,
is consistent with EPA’s determination that no occupational risks
exceed the Agency’s LOC, meaning that even regular occupational
exposures associated with this active ingredient pose negligible risk.

E. Cumulative Exposure 

No mechanism of toxicity in mammals has been identified for Bacillus
subtilis. Therefore, no cumulative effect with other related organisms
is anticipated. Because the available data demonstrate a lack of
toxicity/pathogenicity potential for Bacillus subtilis, adverse dietary
effects are unlikely.

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

Based on the acute toxicity information discussed in section C, Certis
USA believes that the Agency will conclude that there is a reasonable
certainty that no harm will result to the United States population,
including infants and children, from aggregate exposure to residues of
Bacillus subtilis. This includes all anticipated dietary exposures and
all other exposures for which there is reliable information. This
conclusion is based on the data available on Bacillus subtilis which
demonstrate a lack of toxicity/ pathogenicity potential. Thus, there are
no threshold effects of concern and, as a result, Certis USA concludes
that the additional tenfold margin of safety for infants and children is
unnecessary in this instance. Further, the need to consider consumption
patterns, special susceptibility, and cumulative effects does not arise
when dealing with pesticides with no demonstrated significant adverse
effects. 

G. Effects on the Immune and Endocrine Systems 

Bacillus subtilis is a ubiquitous organism in the environment that is
non-toxic to mammals. To date, there is no evidence to suggest that
Bacillus subtilis affects the immune system, functions in a manner
similar to any known hormone, or that it acts as an endocrine disruptor.
Indeed, the submitted toxicity/ pathogenicity studies in rodents
indicate that, following several routes of exposure, the immune system
is intact and able to process and clear Bacillus subtilis. Therefore, it
is unlikely that this organism will have estrogenic or endocrine
effects.  

H. Existing Tolerances

There currently is no existing US exemption from the requirement of a
tolerance for residues of Bacillus subtilis CX-9060. 

I. International Tolerances