Document ID: EPA-HQ-OPP-2003-0308-0010
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2003-10-29T05:00Z

June
20,
2003
Memorandum
Subject:
Review
of
Administrative
Materials
in
support
of
an
Application
for
an
Experimental
Use
Permit
for
Genetically
Modified
Corn
(
Vector
ZMIR39)
Producing
a
Cry3Bb1
Protein
that
Provides
Control
of
Corn
Rootworm
(
Diabrotica
sp.).
EPA
Reg.
No.
524­
EUP­
OA;
DP
Barcode
D287619;
Case
No.
071318;
Submission
No.
S625460;
Chemical
No.
6498.

To:
Mike
Mendelsohn,
Regulatory
Action
Leader
Biopesticides
and
Pollution
Prevention
Division,
7511C
From:
Robyn
Rose,
Entomologist
Biopesticides
and
Pollution
Prevention
Division,
7511C
Peer
Review:
Chris
Wozniak,
Ph.
D.,
Biologist
Biopesticides
and
Pollution
Prevention
Division,
7511C
Classification:
It
is
acceptable
to
plant
corn
containing
ZMIR39
Background:

On
November
11,
2002
Monsanto
submitted
a
request
for
a
FIFRA
Section
G
Experimental
Use
Permit
(
EUP)
for
Cry3Bb1
corn
rootworm
(
CRW,
Diabrotica
spp.)­
protected
corn
events
produced
from
vector
ZMIR39.
Under
this
EUP,
Monsanto
intends
to
plant
ZMIR39
corn
during
the
2003
and
2004
growing
seasons
in
diverse
agronomic
zones
of
25
states
and
Puerto
Rico.
These
studies
are
proposed
to
be
conducted
at
various
research
stations,
seed
production
sites,
privately
owned
and
University
farms
from
the
present
through
February
28,
2005.
Monsanto
intends
to
plant
a
total
of
829
acres
during
the
2003
growing
season
and
2299
acres
during
the
2004
growing
season
(
Table
1).

No
more
than
450
acres
of
ZMIR39
corn
will
be
planted
in
a
state
during
a
growing
season.
A
supplemental
submission
dated
May
19,
2003
details
the
acreage
of
ZMIR39
corn
that
will
be
planted
per
state
for
each
research
trial.
In
addition
to
conducting
studies
with
CRW­
protected
corn
(
vector
ZMIR39),
hybrids
of
ZMIR39
corn
cross­
bred
with
event
MON
810
corn
borerprotected
corn
(
YieldGard
®
)
will
be
tested
under
this
EUP.
The
ZMIR39
corn
expresses
the
Cry3Bb1
protein
and
whereas,
event
MON
810
corn
expresses
the
Cry1Ab
protein.
2
Monsanto
currently
has
a
registration
for
Cry3Bb1
corn
rootworm­
protected
corn
event
MON
863
(
vector
ZMIR13L).
There
is
already
a
tolerance
exemption
granted
for
the
Cry3Bb1
protein
(
40
CFR
§
180.1214);
therefore,
residue
data
is
not
required.
According
to
Monsanto,
event
MON
863
and
vector
ZMIR39
corn
share
an
amino
acid
sequence
identity
of
99.8%;
there
is
only
one
difference
of
the
653
amino
acids.
Differences
between
MON
863
and
ZMIR39
corn
are
the
selectable
marker
proteins
which
are
considered
inert
ingredients.
MON
863
corn
contains
the
nptII
protein
(
neomycin
phosphotransferase
II)
and
ZMIR39
events
contain
5­
enolpyruvylshikimate­
3­
phosphate
synthase
protein
from
Agrobacterium
sp.
strain
CP4
(
CP4
EPSPS).
The
marker
protein
CP4
EPSPS
in
ZMIR39
corn
provides
tolerance
to
glyphosate
herbicide
and
has
been
granted
a
tolerance
(
40
CFR
§
180.1174).

Studies
submitted
in
support
of
the
MON
863
registration
indicate
a
lack
of
toxicity
of
the
Cry3Bb1
protein
to
humans
as
well
as
a
lack
of
risk
to
the
environment.
A
summary
of
these
studies
can
be
found
in
MRID
454240­
09
and
the
Cry3Bb1
Biopesticides
Registration
Action
Document
(
BRAD).
Based
on
results
of
these
previously
submitted
studies,
Monsanto
expects
minimal
risk
from
field­
testing
ZMIR39
corn
and
similar
efficacy
between
MON
863
and
ZMIR39
corn.
None
of
the
ZMIR39
corn
grown
will
be
used
for
commercial
food
or
feed
until
FDA
has
completed
their
safety
and
nutritional
assessment.
ZMIR39
corn
may
be
fed
to
animals
as
part
of
experimental
trials,
otherwise,
the
crop
will
be
destroyed
at
the
end
of
the
trial
or
after
harvest
due
to
USDA­
APHIS
requirements.
Administrative
materials
and
data
were
submitted
in
support
of
this
EUP.
The
information
submitted
as
part
of
the
administrative
materials
are
summarized
and
reviewed
below.

Table
1.
Proposed
Acreage
and
Active
Ingredient
Quantities
for
Each
State
and
Territory
State
Acres
Planting
Seed
(
lb)
A.
I.
in
Planting
Seed
(
g)

2003
2004
Total
2003
2004
Total
2003
2004
Total
AL
2.5
2.5
5
50
50
100
1.36
1.36
2.72
CA
50
450
500
1000
9000
10000
27.24
245.16
272.40
CO
10
20
30
200
400
600
5.45
10.9
16.34
HI
266
616
882
5320
121320
17640
144.92
335.60
480.51
IA
101.2
330.2
431.4
2024
6604
8628
55.13
179.89
235.03
IL
116.5
236
352.5
2330
4720
7050
63.47
128.57
192.04
IN
38.3
85.8
124.1
766
1716
2482
20.87
46.74
67.61
KS
19
36.5
54.5
360
730
1090
9.81
19.89
29.69
KY
0.5
0.5
1
10
10
20
0.27
0.27
0.54
MD
1.3
1.8
3.1
26
36
62
0.71
0.98
1.69
MI
2.5
13
15.5
50
260
310
1.36
7.08
8.44
MN
31.6
42.1
73.7
632
842
1474
17.22
22.94
40.15
3
Table
1.
Continued
State
Acres
Planting
Seed
(
lb)
A.
I.
in
Planting
Seed
(
g)

2003
2004
Total
2003
2004
Total
2003
2004
Total
MO
20.3
30.8
51.1
406
616
1022
11.06
16.78
27.84
MS
2.5
4.5
7
50
90
140
1.36
2.45
3.81
NC
0.5
0.5
1
10
10
20
0.27
0.27
0.54
NE
50.3
101.3
151.6
1006
2026
3032
27.40
55.19
82.59
NM
0.5
0.5
1
10
10
20
0.27
0.27
0.54
NY
0.5
1
1.5
10
20
30
0.27
0.54
0.82
OH
29.6
40.6
70.2
592
812
1404
16.13
22.12
38.24
PA
3.3
16.3
19.6
66
326
392
1.80
8.88
10.68
PR
29
182
211
580
3640
4220
15.80
99.15
114.95
SD
20
30
50
400
600
1000
10.90
16.34
27.24
TN
12.5
12.5
25
250
250
500
6.81
6.81
13.62
TX
5.5
15.5
21
110
310
420
3.00
8.44
11.44
VA
0.5
0.5
1
10
10
20
0.27
0.27
0.54
WI
15.3
28.3
43.6
306
566
872
8.34
15.42
23.75
Total
829
2299
3128
16574
45974
62548
451.48
1252.33
1703.81
(
0.99
lb)
(
2.76
lb)
(
3.75
lb)

EUP
Label
Statements
Proposed
by
Monsanto:

°
"
FOR
EXPERIMENTAL
USE
ONLY"
°
"
Not
for
sale
to
any
person
other
than
a
participant
or
cooperator
of
the
EPA­
approved
Experimental
Use
Program."
°
"
For
use
only
at
a
site
of
a
participant
or
cooperator
and
in
accordance
with
the
terms
and
conditions
of
the
Experimental
Use
Permit."
°
"
This
package
contains
corn
rootworm­
protected
corn
seed
producing
Cry3Bb1,
and
insecticidal
protein
from
Bacillus
thuringiensis.
This
protein
is
effective
in
controlling
feeding
damage
caused
by
corn
rootworm
larvae
(
Diabrotica
spp.)."
°
"
Active
Ingredient:
Corn­
produced
Bacillus
thuringiensis
Cry3Bb1
protein
and
the
genetic
material
necessary
(
vector
ZMIR39)
for
its
production
in
corn...........
0.006%"
°
"
Inert
Ingredients:
Substance
produced
by
marker
gene
and
the
genetic
material
necessary
for
its
production
in
corn...........<
0.007%."
°
"
The
contents
may
only
be
used
according
to
the
approved
EUP
program.
Cooperators
must
have
at
least
one
copy
of
each
applicable
protocol
prior
to
initiating
any
research
4
with
the
contents."
°
"
Any
seed
not
used
in
these
experiments
must
be
returned
to
Monsanto
Company
or
seed
company
partners
/
affiliates,
or
disposed
of
as
specified
in
the
field
protocols."

BPPD
Conclusions:

Insect
Resistance
Management
An
insect
resistance
management
(
IRM)
plan
is
not
necessary
for
the
small
acreage
of
ZMIR39
corn
that
will
be
planted
under
this
EUP.
However,
an
IRM
plan
will
be
required
for
a
Section
3
commercial
registration
for
all
corn
events
containing
the
ZMIR39
vector.

Ecological
Effects
(
Nontarget
Organisms
and
Fate
in
the
Soil)
Adverse
effects
to
populations
of
nontarget
organisms
from
planting
ZMIR39
corn
are
not
expected
during
the
short
duration
and
on
the
limited
acreage
that
will
be
planted
under
this
EUP.
Based
on
previously
submitted
studies
for
registered
Bt
proteins
including
Cry3Bb1,
detectable
amounts
are
not
expected
to
accumulate
and
persist
in
soil
during
this
EUP.
The
following
studies
will
need
to
be
submitted
for
a
Section
3
commercial
registration
for
all
corn
events
containing
the
ZMIR39
vector.

Avian
A
six
week
chicken
broiler
study
with
the
chickens
fed
starter
diets
to
day
17,
grower
diets
to
day
31
and
finisher
diets
to
day
42
(
50­
60
%
transgenic
corn)
should
be
conducted
and
submitted
for
EPA
to
review.
A
control
study
with
non­
transgenic
corn
in
the
standard
diets
should
be
run
concurrently.

Mammalian
Wildlife
Acute
oral
toxicity
including
but
not
limited
to
in
vitro
digestibility,
in
vitro
simulated
gastric
fluid
digestibility
and
a
SDS­
PAGE
sensitivity
analysis
in
support
of
the
simulated
gastric
fluid
digestion
study
will
be
considered
as
part
of
the
risk
assessment
for
mammalian
wildlife
toxicity.

Non­
target
Plants
Since
the
active
ingredient
in
this
product
is
an
insect
toxin
that
has
never
shown
any
toxicity
to
plants,
these
studies
have
been
waived
for
this
product.
In
addition,
there
is
no
evidence
of
transfer
of
any
gene,
including
introduced
genes,
to
organisms
other
than
those
with
which
corn
is
sexually
compatible.
In
addition,
there
are
no
weedy
relatives
of
corn
in
the
continental
U.
S
or
Puerto
Rico.

Non­
target
invertebrates
°
Honey
bee
larval
acute
toxicity
studies
should
be
conducted
and
submitted
to
the
Agency
5
for
review
prior
to
a
full
commercial
registration.
This
study
should
be
conducted
through
adult
emergence.
°
Tier
1
acute
toxicity
tests
should
also
be
conducted
on
three
non­
target
beneficial
invertebrates.
Testing
lady
beetles
(
Coleomegilla
maculata),
the
minute
pirate
bug
(
Orius
insidiosus),
and
a
Carabid
(
e.
g.,
Pterostichus
sp.)
is
recommended.
Since
ZMIR39
is
active
against
coleopterans,
lady
beetle
testing
with
pollen
and
purified
protein
is
recommended.
°
In
addition,
an
appropriate
non­
target
coleopteran
(
e.
g.,
Tetraopes,
milkweed
beetle)
that
may
be
unintentionally
exposed
to
ZMIR39
should
be
evaluated.
°
Field
studies
should
also
be
conducted
that
evaluate
potential
effects
of
growing
ZMIR39
corn
on
varying
trophic
levels
(
e.
g.,
plant
feeders,
predators,
parasitoids,
decomposers).
°
Although
an
earthworm
study
is
not
required
for
full
commercial
registration,
the
Agency
recognizes
their
importance
as
decomposers
living
in
the
soil.
Therefore,
conducting
an
earthworm
dietary
toxicity
study
is
recommended
prior
to
commercialization.
°
A
collembola
study
should
also
be
submitted.

Fish
The
requirement
for
a
static
renewal
toxicity
study
for
freshwater
fish
is
normally
waived
based
on
a
lack
of
exposure
of
fish
to
Bt
proteins
produced
in
corn.
Exposure
from
corn
pollen,
if
it
does
take
place,
will
be
of
a
very
short
duration
and
quantity
and
is
not
expected
to
have
any
detectable
effect
on
freshwater
fish.
A
waiver
request
should
be
submitted
if
nontarget
fish
studies
are
not
conducted.

Aquatic
Invertebrates
Published
and
in­
house
data
for
Bt
crystal
protein
containing
corn
show
that
Bt
crystal
proteins
have
no
measurable
deleterious
effects
on
Daphnia
magna,
a
sensitive
aquatic
invertebrate
species.
Therefore,
no
adverse
effects
are
expected
to
occur
to
nontarget
aquatic
invertebrate
during
the
two
seasons
ZMIR39
will
be
grown
under
this
EUP.
However,
acceptable
Tier
1
toxicity
tests
are
required
for
a
Section
3
commercial
registration.

Estuarine
and
Marine
Animals
The
estuarine
fish
and
shrimp
studies
are
not
required
for
this
product
because
of
very
low
potential
for
exposure.

Endangered
Species
Based
on
the
previously
submitted
Cry
protein
toxicity
and
exposure
data
there
will
not
be
a
"
may
effect"
situation
for
endangered
mammals,
birds,
plants
and
aquatic
species.
All
endangered/
threatened
insect
species
that
are
susceptible
to
Cry3Bb1
(
e.
g.,
coleopterans)
may
be
adversely
affected
if
exposed.
In
the
case
of
Cry3Bb1
protein,
the
test
plots
for
the
proposed
6
EUP
program
must
not
be
near
the
habitats
of
endangered
beetle
(
coleopteran)
species
since
these
may
be
adversely
affected
if
exposed
to
the
Bt
protein
in
the
soil
or
through
pollen
consumption.
An
assessment
of
potential
impacts
on
endangered
species
should
be
conducted
and
submitted
to
the
Agency
for
review
prior
to
full
commercial
registration.

Soil
Fate
The
potential
accumulation
and/
or
persistence
of
Cry3Bb1
expressed
in
ZMIR39
corn
in
soil
should
be
evaluated
prior
to
full
commercial
registration.
This
should
be
done
by
determining
the
DT
50
or
time
to
50%
degradation
of
protein
in
varying
soil
types.
In
particular
the
DT
50
in
a
"
worst
case
scenario"
should
be
determined.
Since
Cry
proteins
have
been
shown
to
readily
bind
to
clay
soils
and
persist
longer
at
low
pH,
the
DT
50
of
Cry3Bb1
should
be
determined
in
soils
with
a
high
clay
content.
Studies
should
include
insect
bioassays
with
a
sensitive
species
such
as
the
Colorado
potato
beetle
as
well
as
an
ELISA
for
comparative
purposes.

Additional
field
studies
should
be
conducted
after
full
commercial
registration.
Field
studies
should
be
conducted
in
a
variety
of
soil
types
and
should
utilize
fields
that
have
had
ZMIR39
corn
grown
for
three
consecutive
years.
Persistence
of
Cry3Bb1
in
the
soil
should
be
monitored
close
to
the
soil
surface
(
e.
g.,
12
­
18
inches
deep)
as
well
as
deep
in
the
soil
(
e.
g.,
several
feet
below
surface
level)
where
microbial
activity
is
reduced.
Sampling
should
occur
immediately
after
anthesis
as
well
as
post­
harvest
and
should
continue
until
no
Bt
protein
is
detected.

Experimental
Program
Summary:

Sampling
Samples
including
whole
corn
plants,
tissue
and/
or
seed
may
be
sent
to
Monsanto
or
other
laboratories
for
analysis.

Isolation
Research
plots
will
be
isolated
from
other
corn
plots
by
planting
Bt
corn

660
ft
from
openpollination
non­
transgenic
corn,
bagging
tassels
from
pre­
anthesis
until
after
pollen
shed
and/
or
detasseling
prior
to
anthesis.

Schedule
There
will
be
continuos
plantings
in
Hawaii
and
Puerto
Rico
with
at
least
60
days
after
harvest
that
fields
will
remain
fallow
and
be
irrigated.
There
will
be
at
least
two
plantings
per
site
in
the
continental
U.
S.
Plantings
in
the
continental
U.
S.
will
occur
from
February
until
June
and
fields
will
be
harvested
from
August
to
November.

Harvest
Procedures
Corn
ears
harvested
by
hand
will
be
put
into
cloth
or
mesh
bags
that
prevent
seed
loss.
Handharvested
ears
may
be
dried
before
shelling
to
remove
moisture.
Seed
from
machine­
harvested
ears
will
be
shelled
during
harvest
and
weighed
prior
to
disposal.
Before
leaving
research
plots,
7
all
equipment
will
be
thoroughly
cleaned.

Final
Disposition
Seed
that
is
not
planted
under
this
EUP
may
be
used
for
other
research
purposes,
as
food
or
feed
if
approved
by
the
FDA,
returned
to
Monsanto
or
seed
companies
based
on
USDA
requirements
or
destroyed
(
e.
g.,
incineration,
autoclaving
or
buried
at
test
sites).
Harvested
corn
may
be
used
in
additional
research,
for
future
plantings
or
as
food
or
feed
in
experimental
studies
or
if
approved
by
the
FDA.
Corn
plants
will
be
destroyed
after
test
termination.

Volunteer
Plants
Plots
and
border
rows
will
only
be
monitored
for
volunteer
plants
in
areas
allowed
to
flower.
Fields
not
allowed
to
flower
will
not
be
monitored
for
volunteer
plants.
All
volunteer
plants
will
be
removed
and
destroyed
prior
to
seed
set
or
flowering.
Monsanto
expects
the
occurrence
of
volunteer
plants
to
be
minimal
because
Bt
plants
will
be
located
in
a
defined
area
and
all
seed
and
plant
material
will
be
appropriately
disposed
of
(
see
Final
Disposition
section
above).
Research
plots
will
be
monitored
for
volunteer
plants
for
at
least
12
months
after
harvest
and
will
continue
until
no
volunteers
are
detected.
If
volunteer
plants
persist,
Cooperators
should
contact
Monsanto.
Plots
in
Hawaii
and
Puerto
Rico
will
be
monitored
for
volunteer
plants
during
the
60
day
fallow
period
after
harvest.

Breeding
and
Observation
Nursery
Trials
According
to
Monsanto,
these
studies
will
be
conducted
to
"[
o]
bserve
the
phenotype
of
corn
expressing
the
cry3Bb1
gene
in
various
elite
corn
lines
derived
from
CRW­
protected
corn
(
vector
ZMIR39)
and/
or
maintain
these
transgenic
lines
through
seed
production."
Corn
plants
containing
vector
ZMIR39
in
various
genetic
backgrounds
will
be
used
in
these
trials
(
Table
2).
Hybrids
containing
vector
XMIR39
and
event
MON
810
will
also
be
planted
(
Table
2).
Plants
containing
the
cry3Bb1
gene
will
be
hand­
pollinated
with
non­
Bt
corn
or
other
Bt
corn
hybrids.
Hand
pollinations
will
include
selfing,
backcrossing
or
crossing
plants
to
a
tester
line.
Plants
may
also
be
sampled
to
determine
phenotype
or
genotype.

Table
2.
Acreage
planted
per
state
in
2003
and
2004
for
the
Breeding
and
Observation
Nursery
Trials
State
ZMIR39
Acreage
2003
Stacked1
Acreage
2003
ZMIR39
Acreage
2004
Stacked1
Acreage
2004
CA
37.5
12.5
CO
1.5
0.5
3.0
1.0
HI
112.5
37.5
150.0
50.0
IA
15.0
5.0
37.5
12.5
IL
25.5
8.5
22.5
7.5
IN
3.8
1.3
3.8
1.3
KS
2.3
0.8
2.3
0.8
8
MI
0.4
0.1
0.8
0.3
MN
2.3
0.8
3.0
1.0
Table
2.
Continued
State
ZMIR39
Acreage
2003
Stacked1
Acreage
2003
ZMIR39
Acreage
2004
Stacked1
Acreage
2004
MO
2.3
0.8
2.3
0.8
NE
2.3
0.8
7.5
2.5
OH
2.3
0.8
3.8
1.3
PA
0.8
0.3
1.5
0.5
PR
11.3
3.8
80.3
26.8
SD
1.5
0.5
2.3
0.8
TN
1.5
0.5
1.5
0.5
TX
0.8
0.3
1.5
0.5
WI
1.5
0.5
3.0
1.0
Total
187.1
62.4
363.8
121.3
1Stacked
products
contain
CRW­
protected
vector
ZMIR39
corn
and
YieldGard
event
MON
810
corn
Inbred
Seed
Increase
Production
Trials
According
to
Monsanto,
these
studies
will
be
conducted
to
"[
p]
roduce
seed
and
plant
material
for
further
testing
and
product
development."
These
trials
will
utilize
a
variety
of
inbred
corn
lines
containing
the
cry3Bb1
gene
(
Table
3).
Corn
plants
will
be
allowed
to
open
pollinate
to
generate
seed
in
plots
that
are
isolated
according
to
the
Isolation
section
above.
Only
corn
containing
vector
ZMIR39
in
various
genetic
backgrounds
will
be
planted.

Table
3.
Acreage
planted
per
state
in
2003
and
2004
for
the
Inbred
Seed
Production
Trials
State
ZMIR39
Acreage
2003
Stacked1
Acreage
2003
ZMIR39
Acreage
2004
Stacked1
Acreage
2004
CA
18.8
6.3
175.0
175.0
HI
37.5
12.5
125.0
125.0
IA
3.8
1.3
15.0
15.0
IL
2.3
0.8
10.0
10.0
IN
1.5
0.5
1.0
1.0
KS
1.5
0.5
1.0
1.0
MN
1.5
0.5
1.0
1.0
9
MO
1.5
0.5
1.0
1.0
NE
1.5
0.5
4.0
4.0
OH
0.8
0.3
1.0
1.0
Table
3.
Continued
State
ZMIR39
Acreage
2003
Stacked1
Acreage
2003
ZMIR39
Acreage
2004
Stacked1
Acreage
2004
PR
4.5
1.5
25.0
25.0
SD
0.8
0.3
0.5
0.5
TN
0.8
0.3
0.5
0.5
WI
0.8
0.3
0.5
0.5
Total
77.3
25.8
360.5
360.5
1Stacked
products
contain
CRW­
protected
vector
ZMIR39
corn
and
YieldGard
event
MON
810
corn
Line
Per
Se
and
Hybrid
Yield
Trials
According
to
Monsanto,
these
studies
will
be
conducted
to
"[
a]
ssess
any
agronomic
effects
of
the
genetic
insertion
in
CRW­
protected
corn
(
vector
ZMIR39)
in
various
corn
genetic
backgrounds."
Only
corn
containing
vector
ZMIR39
in
various
genetic
backgrounds
will
be
planted
(
Table
4).
Inbred
and
hybrid
corn
lines
represented
by
a
segregant
with
and
without
the
cry3Bb1
gene
will
be
planted
in
a
randomized
design
or
as
single
large
blocks.
Up
to
100
different
lines
may
be
planted
at
each
location.
Agronomic
characteristics
such
as
early
plant
vigor,
color,
height,
days
to
flowering
and
maturity,
yield
and
weight
will
be
evaluated.
More
than
one
trial
conducted
at
a
location
will
be
analyzed
as
unique
locations.

Table
4.
Acreage
planted
per
state
in
2003
and
2004
for
the
Line
Per
Se
and
Hybrid
Yield
Trials
State
ZMIR39
Acreage
2003
Stacked1
Acreage
2003
ZMIR39
Acreage
2004
Stacked1
Acreage
2004
CA
18.8
6.3
25.0
25.0
CO
1.5
0.5
3.5
3.5
HI
30.0
10.0
50.0
50.0
IA
28.5
9.5
95.0
95.0
IL
33.8
11.3
64.0
64.0
IN
7.5
2.5
5.0
5.0
KS
4.5
1.5
8.5
8.5
MI
0.8
0.3
4.0
4.0
MN
7.5
2.5
7.5
7.5
MO
4.5
1.5
6.0
6.0
10
MS
0.8
0.3
1.0
1.0
NE
22.5
7.5
17.5
17.5
OH
9.0
3.0
9.0
9.0
PA
0.8
0.3
4.0
4.0
Table
4.
Continued
State
ZMIR39
Acreage
2003
Stacked1
Acreage
2003
ZMIR39
Acreage
2004
Stacked1
Acreage
2004
PR
6.0
2.0
12.5
12.5
SD
2.3
0.8
3.5
3.5
TN
3.0
1.0
2.0
2.0
TX
0.8
0.3
2.5
2.5
WI
3.0
1.0
7.0
7.0
Total
185.3
61.8
327.5
327.5
1Stacked
products
contain
CRW­
protected
vector
ZMIR39
corn
and
YieldGard
event
MON
810
corn
Insect
Efficacy
Trials
According
to
Monsanto,
these
studies
will
be
conducted
to
"[
a]
ssess
the
efficacy
of
corn
expressing
the
cry3Bb1
gene
in
various
corn
genetic
backgrounds
against
corn
rootworm
and
other
potential
soil
pests
of
corn."
Only
corn
containing
vector
ZMIR39
in
various
genetic
backgrounds
will
be
planted
(
Table
5).
These
trials
will
utilize
corn
lines
containing
the
cry3Bb1
gene
planted
in
a
randomized
design
or
as
single
large
blocks.
Control
plots
will
include
nontransgenic
corn
plants
untreated
or
treated
with
insecticides.
More
than
one
trial
conducted
at
a
location
will
be
analyzed
as
unique
locations.

Table
5.
Acreage
planted
per
state
in
2003
and
2004
for
the
Insect
Efficacy
Trials
State
ZMIR39
Acreage
2003
Stacked1
Acreage
2003
ZMIR39
Acreage
2004
Stacked1
Acreage
2004
AL
1.5
1.0
1.3
1.3
CO
2.4
1.6
3.0
3.0
IA
7.9
5.3
10.1
10.1
IL
7.5
5.0
10.5
10.5
IN
3.2
2.1
7.9
7.9
KS
1.8
1.2
5.3
5.3
KY
0.3
0.2
0.3
0.3
MD
0.8
0.5
0.9
0.9
11
MI
0.6
0.4
2.0
2.0
MN
1.6
1.0
2.6
2.6
MO
2.0
1.3
3.9
3.9
MS
0.9
0.6
1.3
1.3
NC
0.3
0.2
0.3
0.3
Table
5.
Continued
State
ZMIR39
Acreage
2003
Stacked1
Acreage
2003
ZMIR39
Acreage
2004
Stacked1
Acreage
2004
NE
3.2
2.1
4.2
4.2
NM
0.3
0.2
0.3
0.3
NY
0.3
0.2
0.5
0.5
OH
3.4
2.2
3.8
3.8
PA
0.8
0.5
3.2
3.2
SD
2.4
1.6
4.5
4.5
TN
1.5
1.0
1.3
1.3
TX
0.9
0.6
2.8
2.8
VA
0.3
0.2
0.3
0.3
WI
2.0
1.3
2.2
2.2
Total
45.7
30.5
71.9
71.9
1Stacked
products
contain
CRW­
protected
vector
ZMIR39
corn
and
YieldGard
event
MON
810
corn
Product
Characterization
and
Performance
Trials
According
to
Monsanto,
these
studies
will
be
conducted
"[
t]
o
produce
grain
and
plant
material
for
product
characterization,
further
regulatory
testing,
and
benefits
studies."
Only
corn
containing
vector
ZMIR39
in
various
genetic
backgrounds
will
be
planted
(
Table
6).
These
trials
will
include
plants
that
are
hand
or
open
pollinated
to
generate
seed
or
plant
tissue
to
be
analyzed
for
composition,
equivalency,
phenotype
and
molecular
characterization.
Only
corn
containing
vector
ZMIR39
in
various
genetic
backgrounds
will
be
planted.

Table
6.
Acreage
planted
per
state
in
2003
and
2004
for
the
Product
Characterization
and
Performance
Trials
State
ZMIR39
Acreage
2003
Stacked1
Acreage
2003
ZMIR39
Acreage
2004
Stacked1
Acreage
2004
CO
1.0
1.0
1.0
1.0
HI
13.0
13.0
33.0
33.0
IA
5.0
5.0
2.0
2.0
12
IL
5.0
5.0
2.5
2.5
IN
1.5
1.5
1.5
1.5
KS
0.5
0.5
0.5
0.5
MN
0.5
0.5
0.5
0.5
MO
0.5
0.5
0.5
0.5
NE
0.5
0.5
1.5
1.5
Table
6.
Continued
State
ZMIR39
Acreage
2003
Stacked1
Acreage
2003
ZMIR39
Acreage
2004
Stacked1
Acreage
2004
OH
0.5
0.5
0.5
0.5
SD
0.5
0.5
0.5
0.5
TN
0.5
0.5
0.5
0.5
WI
0.5
0.5
0.5
0.5
Total
29.5
29.5
45.0
45.0
1Stacked
products
contain
CRW­
protected
vector
ZMIR39
corn
and
YieldGard
event
MON
810
corn
Insect
Resistance
Management
Trials
According
to
Monsanto,
these
studies
will
be
conducted
"[
t]
o
develop
additional
data
for
use
in
further
refining
a
corn
rootworm
insect
resistance
management
(
IRM)
plan.
Studies
include
characterizing
dose,
larval
movement,
adult
movement
and
mating
behavior,
and
effects
of
the
Cry3Bb1
protein
on
adult
CRW
fitness."
Only
corn
containing
vector
ZMIR39
in
various
genetic
backgrounds
will
be
planted
(
Table
7).
These
trials
will
utilize
corn
lines
containing
the
cry3Bb1
gene
planted
in
a
randomized
design
or
as
single
large
blocks.
Control
plots
will
include
nontransgenic
corn
plants
untreated
or
treated
with
insecticides.
Efficacy
trials
as
well
as
CRW
adult
emergence
and
fitness
studies
will
be
conducted.
Across
and
between
row
larval
movement
will
also
be
evaluated
in
Bt
and
non­
Bt
plots.
In
addition,
various
ratios
of
Bt
and
non­
Bt
corn
seed
mixtures
will
be
evaluated.
Yields
for
each
treatment
will
also
be
determined.

Table
7.
Acreage
planted
per
state
in
2003
and
2004
for
the
Insect
Resistance
Management
Trials
State
ZMIR39
Acreage
2003
Stacked1
Acreage
2003
ZMIR39
Acreage
2004
Stacked1
Acreage
2004
CO
0.5
0.5
IA
4.0
4.0
4.0
4.0
IL
2.5
2.5
2.5
2.5
IN
5.0
5.0
12.5
12.5
KS
1.0
1.0
1.0
1.0
13
MN
5.0
5.0
5.0
5.0
MO
2.0
2.0
2.0
2.0
NE
2.0
2.0
5.0
5.0
OH
2.0
2.0
2.0
2.0
SD
2.0
2.0
2.0
2.0
TN
0.5
0.5
0.5
0.5
TX
0.5
0.5
1.0
1.0
Table
7.
Continued
State
ZMIR39
Acreage
2003
Stacked1
Acreage
2003
ZMIR39
Acreage
2004
Stacked1
Acreage
2004
WI
1.0
1.0
1.0
1.0
Total
27.5
27.5
39.0
39.0
1Stacked
products
contain
CRW­
protected
vector
ZMIR39
corn
and
YieldGard
event
MON
810
corn
Nontarget
Organisms
and
Benefits
Trials
According
to
Monsanto,
these
studies
will
be
conducted
to
"[
d]
evelop
additional
data
to
assess
the
impact
of
the
Cry3Bb1
protein
on
nontarget
organisms,
including
a
wide
variety
of
soil
and
surface­
dwelling
organisms."
Only
corn
containing
vector
ZMIR39
in
various
genetic
backgrounds
will
be
planted.
These
trials
will
utilize
corn
lines
containing
the
cry3Bb1
gene
planted
in
a
randomized
design
or
as
single
large
blocks
in
a
variety
of
environments.
Control
plots
will
include
non­
transgenic
corn
plants
untreated
or
treated
with
insecticides.
The
amount
of
Cry3Bb1
protein
in
Bt
and
non­
Bt
plots
will
be
evaluated
as
well
as
microbial
community
structure,
earthworm
populations
and
terrestrial
invertebrates
at
and
below
the
soil
surface.

Table
8.
Acreage
planted
per
state
in
2003
and
2004
for
the
Nontarget
Organisms
and
Benefits
Trials
State
ZMIR39
Acreage
2003
Stacked1
Acreage
2003
ZMIR39
Acreage
2004
Stacked1
Acreage
2004
IA
2.5
2.5
3.0
3.0
IL
2.5
2.5
2.5
2.5
IN
1.5
1.5
2.5
2.5
KS
0.5
0.5
0.5
0.5
MN
1.5
1.5
2.5
2.5
MO
0.5
0.5
0.5
0.5
NE
1.5
1.5
2.5
2.5
OH
1.0
1.0
1.0
1.0
14
SD
2.0
2.0
2.0
2.0
TN
0.5
0.5
0.5
0.5
WI
0.5
0.5
0.5
0.5
Total
14.5
14.5
18.0
18.0
1Stacked
products
contain
CRW­
protected
vector
ZMIR39
corn
and
YieldGard
event
MON
810
corn
Seed
Treatment
Trials
According
to
Monsanto,
these
studies
will
be
conducted
"[
t]
o
evaluate
effects
of
diverse
chemical
seed
treatments,
in
combination
with
transgenic
corn,
on
corn
rootworm
and
additional
insect
pests."
Only
corn
containing
vector
ZMIR39
in
various
genetic
backgrounds
will
be
planted
(
Table
9).
Bt
and
non­
transgenic
corn
seed
will
be
treated
with
soil
insecticides
and/
or
insect
deterrents.
Experimental
plots
will
be
planted
in
a
randomized
design
or
as
single
large
blocks
in
a
variety
of
environments.
Control
plots
will
include
non­
transgenic
corn
plants
untreated
or
treated
with
insecticides.
Efficacy
of
the
various
treatments
against
CRW
and
other
invertebrate
pests
will
be
evaluated.
Agronomic
characteristics
(
e.
g.,
germination,
seedling
vigor)
will
also
be
evaluated.

Table
9.
Acreage
planted
per
state
in
2003
and
2004
for
the
Seed
Treatment
Trials
State
ZMIR39
Acreage
2003
Stacked1
Acreage
2003
ZMIR39
Acreage
2004
Stacked1
Acreage
2004
IA
1.0
1.0
1.0
1.0
IL
1.0
1.0
1.0
1.0
NE
1.0
1.0
1.0
1.0
OH
0.5
0.5
0.5
0.5
SD
0.5
0.5
0.5
0.5
TX
0.5
0.5
0.5
0.5
WI
0.5
0.5
0.5
0.5
Total
5.0
5.0
5.0
5.0
1Stacked
products
contain
CRW­
protected
vector
ZMIR39
corn
and
YieldGard
event
MON
810
corn
Swine
Growth
and
Feed
Efficiency
Trials
According
to
Monsanto,
these
studies
will
be
conducted
"[
t]
o
evaluate
feeding
performance
in
growing­
finishing
pigs
fed
diets
containing
grain
from
CRW­
protected
corn
(
vector
ZMIR39)."
These
trials
will
use
half
gilt
and
half
barrow
pigs
which
will
be
put
in
pens
according
to
body
weight
and
allowed
ad
libitum
access
to
food
and
water.
Pigs
will
be
fed
CRW­
protected
corn
(
vector
ZMIR39)
or
a
control
with
similar
genetic
background
or
one
of
two
other
commercial
hybrids
until
they
reach
market
weight
(
Table
10).
Feeding
will
occur
from
harvest
to
approximately
120
days
post
harvest
in
2004
only.
Weight
and
food
intake
will
be
determined
periodically.
Pigs
will
be
returned
to
their
owner
or
sent
to
a
commercial
slaughterhouse
for
meat
15
production.

Isolation
will
be
accomplished
in
the
study
by
using
confined
pens,
grinding
grain
prior
to
feeding
to
devitalize
it
and
monitoring
for
volunteer
plants.
Volunteer
plants
will
be
monitored
as
described
in
the
Volunteer
Plants
section
above.
Corn
not
fed
to
pigs
will
be
destroyed
according
to
USDA
guidelines,
stored
on
site,
used
in
other
research
trials,
used
as
food
or
feed
upon
FDA
approval
or
returned
to
Monsanto.

Table
10.
Acreage
planted
per
state
in
2004
for
the
Swine
Growth
and
Feed
Efficiency
Trials
State
ZMIR39
Acreage
2004
Stacked1
Acreage
2004
IA
2.5
2.5
IL
2.5
2.5
Table
10.
Continued
State
ZMIR39
Acreage
2004
Stacked1
Acreage
2004
IN
2.5
2.5
NE
2.5
2.5
Total
10.0
10.0
1Stacked
products
contain
CRW­
protected
vector
ZMIR39
corn
and
YieldGard
event
MON
810
corn
Dairy
Cattle
Feed
Efficiency
Trials
According
to
Monsanto,
these
studies
will
be
conducted
"[
t]
o
evaluate
milk
yield
and
feeding
performance
in
lactating
dairy
cows
fed
diets
containing
grain
and
silage
from
CRW­
protected
corn
(
vector
XMIR39)."
Lactating
dairy
cattle
will
be
fed
fermented
silage
and
ground
grain
ad
libitum
from
harvest
to
approximately
110
days
post
harvest
in
2004
only.
Cattle
will
be
fed
CRW­
protected
corn
(
vector
ZMIR39)
or
a
control
with
similar
genetic
background
or
one
of
two
other
commercial
hybrids
(
Table
11)..
Treatments
will
be
applied
in
a
latin
square
design
and
feed
intake
and
lactation
performance
will
be
evaluated.

Isolation
will
be
accomplished
by
performing
studies
in
confined
pens
and
fermenting
silage
which
results
in
devitalization.
Volunteer
plants
will
be
monitored
and
removed
prior
to
seed
set
or
flowering
in
feed
bunkers
and
feed
milling
sites
for
at
least
12
months
and
continue
until
no
volunteers
are
found.
Corn
not
fed
to
cattle
will
be
destroyed
according
to
USDA
guidelines,
stored
on
site,
used
in
other
research
trials,
used
as
food
or
feed
upon
FDA
approval
or
returned
to
Monsanto.
Cattle
will
be
returned
to
owners
after
completion
of
the
studies.

Table
11.
Acreage
planted
per
state
in
2004
for
the
Dairy
Cattle
Feed
Efficiency
Trials
State
ZMIR39
Acreage
2004
Stacked1
Acreage
2004
IA
2.5
2.5
IL
2.5
2.5
16
IN
2.5
2.5
NE
2.5
2.5
Total
10.0
10.0
1Stacked
products
contain
CRW­
protected
vector
ZMIR39
corn
and
YieldGard
event
MON
810
corn
Beef
Cattle
Growth
and
Feed
Efficiency
According
to
Monsanto,
these
studies
will
be
conducted
"[
t]
o
evaluate
growth
and
feeding
performance
in
feedlot
cattle
fed
diets
containing
grain
from
CRW­
protected
corn
(
vector
ZMIR39)."
Steers
assigned
to
pens
according
to
weight
will
be
fed
CRW­
protected
corn
(
vector
ZMIR39)
or
a
control
with
similar
genetic
background
or
one
of
two
other
commercial
hybrids
ad
libitum
(
Table
12).
Testing
will
take
place
until
steers
reach
market
weight
which
will
occur
from
harvest
until
approximately
100
to
150
days
post
harvest
in
2004
only.
Feed
will
include
ground
corn
grain
and
intake
and
weight
will
be
recorded
periodically
during
testing.

Isolation
will
be
accomplished
by
performing
studies
in
confined
areas
and
grinding
corn
which
results
in
devitalization.
Volunteer
plants
will
be
monitored
and
removed
prior
to
seed
set
or
flowering
in
feed
bunkers
and
feed
milling
sites
for
at
least
12
months
and
continue
until
no
volunteers
are
found.
Corn
not
fed
to
steers
will
be
destroyed
according
to
USDA
guidelines,
stored
on
site,
used
in
other
research
trials,
used
as
food
or
feed
upon
FDA
approval
or
returned
to
Monsanto.
Cattle
will
be
returned
to
their
owner
or
sent
to
a
commercial
slaughterhouse
for
meat
production.

Table
12.
Acreage
planted
per
state
in
2004
for
the
Dairy
Cattle
Feed
Efficiency
Trials
State
ZMIR39
Acreage
2004
Stacked1
Acreage
2004
IA
2.5
2.5
IL
2.5
2.5
IN
2.5
2.5
NE
2.5
2.5
Total
10.0
10.0
1Stacked
products
contain
CRW­
protected
vector
ZMIR39
corn
and
YieldGard
event
MON
810
corn
Cattle
Grazing
Feed
Efficiency
Trials
According
to
Monsanto,
these
studies
will
be
conducted
"[
t]
o
evaluate
feeding
performance
of
cattle
as
a
result
of
grazing
on
corn
stover
and
stubble
from
CRW­
protected
corn
(
vector
ZMIR39)."
Calves
or
mature
cattle
will
be
assigned
to
pens
according
to
weight
will
be
fed
CRW­
protected
corn
(
vector
ZMIR39)
or
a
control
with
similar
genetic
background
or
one
of
two
other
commercial
hybrids
ad
libitum.
Test
animals
will
be
allowed
to
freely
graze
on
corn
stover
and
stubble
in
test
plots
after
harvest
for
up
to
approximately
90
days
in
2004
only
(
Table
13).
Body
weight
of
cattle
will
be
monitored
and
recorded.
Fields
used
for
grazing
will
be
monitored
for
volunteer
plants
for
at
least
12
months
and
continue
until
no
volunteers
are
found.
17
Grain
is
expected
to
be
devitalized
after
digestion
by
test
animals.
Corn
not
fed
to
cattle
will
be
destroyed
according
to
USDA
guidelines,
stored
on
site,
used
in
other
research
trials,
used
as
food
or
feed
upon
FDA
approval
or
returned
to
Monsanto.
Cattle
will
be
returned
to
their
owner
at
test
termination.

Table
13.
Acreage
planted
per
state
in
2004
for
the
Dairy
Cattle
Feed
Efficiency
Trials
State
ZMIR39
Acreage
2004
Stacked1
Acreage
2004
IA
2.5
2.5
IL
2.5
2.5
IN
2.5
2.5
NE
2.5
2.5
Total
10.0
10.0
1Stacked
products
contain
CRW­
protected
vector
ZMIR39
corn
and
YieldGard
event
MON
810
corn