Document ID: EPA-HQ-OPP-2006-0300-0005
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2006-05-26T04:00Z

1
EPA
BIOPESTICIDES
AND
POLLUTION
PREVENTION
DIVISION
COMPANY
NOTICE
OF
FILING
TEMPLATE
FOR
PESTICIDE
PETITIONS
PUBLISHED
IN
THE
FEDERAL
REGISTER
(
11/
15/
2005)

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
establish
a
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
food
and
feed
commodities
of
field
corn,
sweet
corn,
and
popcorn.
EPA
is
being
petitioned
to
establish
a
temporary
tolerance
exemption
as
part
of
the
Experimental
Use
Permit
524­
EUP­
OT.

plant­
incorporated
protectant
Bacillus
thuringiensis
Cry2Ab2
protein
and
the
genetic
material
necessary
for
its
production
in
food
and
feed
commodities
of
field
corn,
sweet
corn,
and
popcorn.
EPA
is
being
petitioned
to
establish
a
temporary
tolerance
exemption
as
part
of
the
Experimental
Use
Permit
524­
EUP­
OT.
2
[
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;
EPA
has
not
fully
evaluated
the
merits
of
the
pesticide
petition.
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
wellestablished
human
and
animal
safety.
The
Cry2Ab2
protein
has
been
previously
expressed
in
corn
(
MON
84006)
and
cotton
(
Bollgard
II
®
)
and
the
protein's
safety
has
been
fully
characterized.

[
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.
Data
characterizing
the
Cry1Ab
and
Cry1Ac
proteins
produced
in
corn
and
cotton
have
been
submitted
to
and
reviewed
by
EPA.
Safety
assessment
as
well
as
sequence,
structural,
and
functional
similarity
of
Cry1A.
105
to
Cry1Ac
and
Cry1Ab
proteins
data
are
included
in
the
submission.
The
results
are
consistent
with
the
overall
biochemical
and
functional
characteristics
of
Cry1A
proteins
and
indicate
the
general
safety
of
Cry1A.
105
for
humans.
3
The
Cry2Ab2
protein
is
derived
from
Bacillus
thuringiensis
subsp.
kurstaki
and
has
greater
than
95%
sequence
identity
to
the
wild
type
Cry2Ab.
The
Cry2Ab2
protein
has
been
previously
expressed
in
corn
(
MON
84006)
and
cotton
(
Bollgard
II);
the
protein's
safety
has
been
fully
characterized.
The
Cry2Ab2
safety
assessment
has
been
reviewed
by
EPA.
A
time­
limited
exemption
from
the
requirement
of
a
tolerance
was
issued
for
corn
and
cotton
in
2001
(
40
CFR
§
180.1214)
and
amended
to
remove
time
limitation
for
cotton
in
2004
(
40
CFR
§
180.1215).

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

[
COMMENT
21]
Cry1A.
105
and
Cry2Ab2
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,
analytical
methods
for
detection
of
the
Cry1A
and
Cry2Ab2
proteins
are
available
commercially
and
allow
for
identification
of
ZMIR245
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
40
years.
The
numerous
toxicology
studies
conducted
with
these
microbial
products
show
no
significant
adverse
effects,
and
demonstrate
that
the
products
are
practically
nontoxic
to
mammals.

Data
have
been
submitted
demonstrating
the
lack
of
mammalian
toxicity
at
high
levels
of
exposure
to
purified
Cry1A.
105
and
Cry2Ab2
proteins.
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
studies
have
been
evaluated
for
Cry2Ab2
and
the
Cry1A.
015
proteins.
Cry2Ab2,
Cry1Ab,
and
Cry1Ac
proteins
with
evaluated
safety
have
been
placed
in
toxicity
category
4
IV
due
to
the
lack
of
any
evidence
of
toxicity.
Furthermore,
amino
acid
sequence
comparisons
of
the
Cry1A.
105
and
Cry2Ab2
proteins
to
known
toxic
proteins
showed
no
similarities.

Because
Cry1A.
105
and
Cry2Ab2
are
proteins,
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
and
Cry2Ab2
proteins
are
rapidly
degraded
by
simulated
gastric
fluid
in
vitro.
In
a
solution
of
simulated
gastric
fluid,
complete
degradation
of
the
detectable
Cry1A.
105
and
Cry2Ab2
proteins
occurred
within
30
and
15
seconds
of
incubation,
respectively.

An
analysis
of
amino
acid
sequences
of
known
allergens
uncovered
no
evidence
of
sequence
homology
with
the
Cry1A.
105
or
Cry2Ab2
proteins,
even
at
the
level
of
eight
contiguous
amino
acid
residues.

The
genetic
material
encoding
the
Cry1A.
105
and
Cry2Ab2
proteins
and
the
regulatory
regions
controlling
expression
of
the
cry1A.
105
or
cry2Ab2
genes
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
and
Cry2Ab2
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
and
Cry2Ab2
are
plant­
incorporated
protectants
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
5
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
Cry1Ab,
Cry1Ac,
and
Cry2Ab2
proteins
are
rapidly
degraded
in
soil
and,
therefore,
are
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
plantincorporated
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
and
Cry2Ab2
proteins
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
plantincorporated
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
and
Cry2Ab2
proteins
and
the
genetic
material
necessary
for
their
production
in
corn
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
and
Cry2Ab2
proteins.

[
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
6
provide
no
evidence
of
adverse
threshold
effects
for
the
Cry1A.
105
or
Cry2Ab2
proteins
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
and
Cry2Ab2
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]
Several
Cry1A
proteins
and
the
genetic
material
necessary
for
their
production
in
raw
agricultural
commodities
and
all
plants
are
exempt
from
the
requirement
of
a
tolerance
(
40
CFR
§
180.1155;
40
CFR
§
180.1173).

The
Bacillus
thuringiensis
Cry2Ab2
protein
and
the
genetic
material
necessary
for
its
production
in
cotton
are
exempt
from
the
requirement
of
a
tolerance
when
used
as
a
plant
incorporated
protectant
in
the
food
and
feed
commodities,
cotton
seed,
cotton
oil,
cotton
meal,
cotton
hay,
cotton
hulls,
cotton
forage,
and
cotton
gin
byproducts
(
40
CFR
§
180.1215).

[
COMMENT
52]
I.
International
Tolerances
[
COMMENT
55]
No
Codex
maximum
residue
levels
have
been
established
for
these
plant­
incorporated
protectants
at
this
time.