**EPA BIOPESTICIDES AND POLLUTION PREVENTION DIVISION COMPANY NOTICE OF
FILING TEMPLATE FOR PESTICIDE PETITIONS PUBLISHED IN THE FEDERAL
REGISTER**

**(03/09/2007)**

**EPA Biopesticides and Pollution Prevention Division contact: Mike
Mendelsohn, telephone number: 703-308-8715**

**COMMENT 1\] Monsanto Company**

**\[COMMENT 2\] \[Insert petition number\]**

**\[COMMENT 3\]** EPA has received a pesticide petition **\[insert
petition number\]** from Monsanto Company, 800 North Lindbergh Blvd.,
St. Louis, MO 63167, 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

**\[COMMENT 4\]** NA-Remove

**\[COMMENT 5\]** NA-Remove

**\[COMMENT 6\]** to remove time limitation from the temporary exemption
from the requirement of a tolerance for the

**\[COMMENT 7\]** NA-Remove

**\[COMMENT 8\]** NA-Remove

**\[COMMENT 9\]** plant-incorporated protectant *Bacillus thuringiensis*
Cry1A.105 protein and the genetic material necessary for its production
in all crops and agricultural commodities. EPA is being petitioned to
establish a permanent tolerance exemption as part of the registration of
insect-protected corn MON 89034.

**\[COMMENT 10\]** Pursuant to section 408(d)(2)(A)(i) of the FFDCA, as
amended, Monsanto Company has submitted the following summary of
information, data, and arguments in support of their pesticide petition.
This summary was prepared by Monsanto Company; The summary may have been
edited by EPA to clarify terminology, remove extraneous material, or if
the summary unintentionally made the reader conclude that the findings
reflected EPA's position and not the position of the petitioner.

**\[COMMENT 11\]** I. **Monsanto Company Petition Summary**

**\[COMMENT 12\] \[Insert petition number\]**

**\[COMMENT 13\]** *A. Product name and Proposed Use Practices*

**\[COMMENT 14\]** Corn has been genetically modified to produce
*Bacillus thuringiensis* (*Bt*) Cry1A.105 and Cry2Ab2 proteins. Plants
producing these proteins are derived from transformation events that
contain the insecticidal proteins and the genetic material necessary for
their expression in corn (vector ZMIR245). The combination of Cry2Ab2
and Cry1A.105 proteins provides a broad spectrum of activity against
lepidopteran insect pests and a tool for management of potential insect
resistance. The Cry1A.105 protein is closely related to the Cry1A class
of proteins, which has well-established human and animal safety. Both
the Cry1A.105 and Cry2Ab2 proteins have been fully characterized and no
adverse effects were observed for human health and environmental safety.

**\[COMMENT 15\]** *B. Product Identity/Chemistry*

**\[COMMENT 16\]** 1. **\[COMMENT 17\]** *Identity of the pesticide and
corresponding residues.*

**\[COMMENT 18\]** The Cry1A.105 protein is 93.6, 90.0, and 76.7 %
identical on the amino acid level to the Cry1Ac, Cry1Ab, and Cry1F
proteins, respectively. The gene encoding Cry1A.105 was constructed by
combining the nucleotide sequences that encode domains I and II of
Cry1Ab, domain III of Cry1F, and substantially all of the C-terminal
domain of Cry1Ac. Comparison of the Cry1A.105 sequence to other Cry
proteins on the *Bt* phylogenetic tree demonstrated that Cry1A.105 is
located in the same cluster as Cry1Ac and is also related to Cry1Ab. The
Cry1A.105 protein was fully characterized and equivalence between *in
planta*- and *E. coli*-produced proteins used in the safety assessment
was demonstrated.

**\[COMMENT 19\]** 2. **\[COMMENT 20\]** *Magnitude of residue at the
time of harvest and method used to determine the residue.*

**\[COMMENT 21\]** Cry1A.105 residue data are not required for an
assessment of human health effects because of the lack of mammalian
toxicity.

**\[COMMENT 22\]** 3. **\[COMMENT 23\]** *A statement of why an
analytical method for detecting and measuring the levels of the
pesticide residue is not needed.*

**\[COMMENT 24\]** An analytical method is not required because this
petition requests an exemption from tolerances. However, a validated
ELISA method for detection of Cry1A.105 protein and analytical methods
for detection of the Cry1Ab and Cry2Ab2 proteins are available and allow
for identification of MON 89034 corn.

**\[COMMENT 25\]** *C. Mammalian Toxicological Profile*

Cry proteins have been used safely and effectively as pest control
agents in microbial *Bt* formulations for more than 45 years. The
numerous toxicology studies conducted with these microbial products show
no significant adverse effects, and demonstrate that the products are
nontoxic to mammals.

Data have been submitted demonstrating the lack of mammalian toxicity at
high levels of exposure to purified Cry1A.105 protein. These data
demonstrate the safety of the protein at levels well above maximum
possible exposure levels that are reasonably anticipated in crops. This
conclusion is consistent with the Agency position regarding toxicity and
residue data requirements for the microbial *Bt* products which utilize
the same proteins as these plant-incorporated protectants (40 CFR
§158.740(b)(2)(i)). For microbial products, further toxicity and residue
testing are only triggered by significant acute effects in studies such
as the mouse oral toxicity study.

When proteins are toxic, they are known to act via acute mechanisms and
at very low levels (Sjoblad et al*.* "Toxicological Considerations for
Protein Components of Biological Pesticide Products." *Reg. Toxicol.
Pharmacol.* 15:3-9, 1992). Acute oral toxicity study has been evaluated
for the Cry1A.105 protein. Furthermore, amino acid sequence comparisons
of the Cry1A.105 protein to known toxic proteins showed no similarities.

Because Cry1A.105 is a protein, the potential for allergic sensitivities
was evaluated. Common food allergens are present at high concentrations
in food, are resistant to pepsin digestion, may be resistant to acid or
heat, and can be glycosylated. Data have been submitted demonstrating
that the Cry1A.105 protein is rapidly degraded by simulated gastric
fluid *in vitro*. In a solution of simulated gastric fluid, complete
degradation of the detectable Cry1A.105 protein occurred within 30
seconds of incubation.

An analysis of amino acid sequences of known allergens uncovered no
evidence of sequence homology with the Cry1A.105 protein, even at the
level of eight contiguous amino acid residues.

The genetic material encoding the Cry1A.105 protein and the regulatory
regions controlling expression of the *cry1A.105* gene are nucleic acids
(DNA and RNA). DNA and RNA occur in all forms of plant and animal life
and there is no documented instance of nucleic acids being associated
with toxic effects when consumed as a component of food. DNA and RNA are
in the category of substances generally considered as safe (GRAS). EPA
exempts nucleic acids from the FFDCA Section 408 requirements. Data
characterizing the genetic material necessary for the production of
Cry1A.105 in corn were provided to the Agency. No mammalian toxicity is
anticipated from dietary exposure to the genetic material necessary for
production of the subject plant-incorporated protectant.

**\[COMMENT 26\]** *D. Aggregate Exposure*

**\[COMMENT 27\]** 1. **\[COMMENT 28\]** *Dietary exposure*

**\[COMMENT 29\]** Cry1A.105 is a plant-incorporated protectant in corn,
thus dietary exposure is deemed to be the most relevant route for
assessing human risk.

**\[COMMENT 30\]** i. **\[COMMENT 31\]** *Food*

**\[COMMENT 32\]** Monsanto has considered available information on the
aggregate exposure levels to consumers and major identifiable groups of
consumers to the protein residue and related substances. These
considerations include dietary exposure under the tolerance exemptions
in effect for the plant-incorporated protectant residue, and exposure
from non-occupational sources. Oral exposure may occur at very low
levels from ingestion of processed corn products. However, a lack of
mammalian toxicity and the rapid digestibility of the plant-incorporated
protectants have been demonstrated.

**\[COMMENT 33\]** ii*.***\[COMMENT 34\]** *Drinking water*

**\[COMMENT 35\]** Oral exposure from ingestion of drinking water is
unlikely because the proteins are present at low levels within the plant
and previously submitted studies demonstrate that Cry1A.105 protein is
rapidly degraded in soil and, therefore, is not present in drinking
water.

**\[COMMENT 36\]** 2. **\[COMMENT 37\]** *Non-dietary exposure*

**\[COMMENT 38\]** Exposure via skin or inhalation is not likely because
the plant-incorporated protectants are contained within plant cells,
which essentially eliminates these exposure routes or reduces them to
negligible levels. The use sites for the Cry1A.105 protein are all
agricultural for control of insects. Exposure to workers and bystanders
resulting from plant pesticides will be negligible and unlikely to add
measurably to any exposure resulting from microbial or other *Bacillus
thuringiensis* formulations. Exposure to infants and children via
residential or lawn use is not expected.

**\[COMMENT 39\]** *E. Cumulative Exposure*

**\[COMMENT 40\]** Because there is no indication of mammalian toxicity
to the plant-incorporated protectants there will be no opportunity for
cumulative toxic effects.

**\[COMMENT 41\]** *F. Safety Determination*

**\[COMMENT 42\]** 1. **\[COMMENT 43\]** *U.S. population*

**\[COMMENT 44\]** Sufficient data have been submitted to assess the
health risk of Cry1A.105 protein and the genetic material necessary for
its production in all raw agricultural commodities. The absence of
toxicity in high dose acute oral studies, the lack of sequence homology
with known protein toxins, rapid digestion in a gastric matrix, and
minimal allergenicity potential provide a reasonable certainty of no
harm for the U.S. general population potentially exposed to the
Cry1A.105 protein.

**\[COMMENT 45\]** 2. **\[COMMENT 46\]** *Infants and children*

**\[COMMENT 47\]** Non-dietary exposure to infants and children is not
anticipated due to the patterns of use for these plant-incorporated
protectants. The submitted data provide no evidence of adverse threshold
effects for the Cry1A.105 protein that would warrant application of an
additional safety factor for the protection of infants and children.
Furthermore, the provisions for consumption patterns, special
susceptibility, and cumulative effects do not apply.

**\[COMMENT 48\]** *G. Effects on the Immune and Endocrine Systems*

**\[COMMENT 49\]** The lack of Cry1A.105 homology to known toxins and
allergens and their rapid degradation in a mammalian digestive system
suggests minimal risk for adverse effects on the immune system. These
pesticidal active ingredients are proteins derived from sources that are
not known to exert an influence on the endocrine system.

**\[COMMENT 50\]** *H. Existing Tolerances*

**\[COMMENT 51\]** The *Bacillus thuringiensis* Cry1A.105 protein and
the genetic material necessary for its production in corn is exempt from
the requirement of a tolerance when used as a plant incorporated
protectant in food and feed commodities of field corn, sweet corn, and
popcorn until June 30, 2009 (40 CFR§174.453).

**\[COMMENT 52\]** *I. International Tolerances*

**\[COMMENT 53\]** No Codex maximum residue levels have been established
for these plant-incorporated protectants at this time.