Document ID: EPA-HQ-OPP-2004-0395-0036
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2005-02-16T05:00Z

29
DATA
EVALUATION
REPORT
6/
17/
2001
Reviewed
by:
Chris
A.
Wozniak,
Ph.
D.,
Biologist,
OPP
/
BPPD
/
s/
Secondary
Reviewer:
John
L.
Kough,
Ph.
D.,
Biologist,
OPP
/
BPPD
/
s/
MRID
No.:
452422­
09
Active
Ingredients:
Bacillus
thuringiensis
strain
PS149B1
(
NRRL
B­
21553)
binary
insect
control
proteins
as
expressed
in
maize
Product
Name:
PS149B1
insect
control
proteins
as
expressed
in
maize
ID
No.:
068467­
EUP­
G,
068467­
EUP­
L,
029964­
EUP­
R,
022964­
EUP­
G
Submission
No.:
S590323,
S590337
Chemical
No.:
006430
Bacillus
thuringiensis
PS149B1
proteins
DP
Barcode:
D271578
Sponsor:
Dow
Agrosciences
LLC,
9330
Zionsville
Rd.,
Indianapolis,
IN
46268.
Authors:
K.
J.
Brooks,
P.
M.
DeWildt
Testing
Facility:
Toxicology
and
Environmental
Research
and
Consulting,
The
Dow
Chemical
Company,
Midland,
MI
48674
Study
Titles:
PS149B1
14
kDa
and
44
kDa
proteins:
acute
oral
toxicity
study
in
CD­
1
mice.
Study
Date:
October
10,
2000
Study
No.:
001128
Conclusion:
All
animals
survived
the
two
week
study.
One
female
mouse
exhibited
protruding
or
bulging
eyes
on
days
6
and
7,
but
this
resolved
thereafter.
This
observation
was
not
attributed
to
the
treatment
as
it
was
an
isolated
observation
(
i.
e.,
no
other
animals
exhibited
this).
No
other
clinical
signs
were
noted
for
any
animals
during
the
study.
An
initial
weight
loss
was
observed
in
two
mice
at
test
days
1
and
2,
but
both
gained
weight
for
the
remainder
of
the
study.
All
other
animals
gained
weight
throughout
the
study.
No
gross
treatment
related
observations
were
recorded
during
the
study
as
represented
by
gross
pathologic
observations.
An
acute
oral
LD50
was
calculated
for
this
study
based
upon
a
dosage
of
a
1:
4.6
ratio
mixture
of
PS149B1
proteins
at
greater
than
5000
mg/
kg
and
greater
than
2000
mg/
kg
for
an
equimolar
mixture
(
1:
3)
of
the
pure
proteins.

Classification:
Acceptable.

Good
Laboratory
Practices:
This
study
was
conducted
in
compliance
with
the
Certification
of
Good
Laboratory
Practices
as
outlined
in
40
CFR
160,
with
the
following
exception:
dose
mixtures
were
not
analyzed
for
homogeneity
and
dose
confirmation.

Purpose:
To
determine
the
median
lethal
oral
dose
(
LD50)
of
the
PS149B1
14
kDa
and
44
kDa
proteins.

METHODS
30
Test
material
­
PS149B1
proteins
in
a
ratio
of
1:
4.6
(
14
kDa:
44
kDa).
Supplied
by
Dow
AgroSciences
(
DAS)
PS149B1
14
kDa
­
040400POOL,
44
kDa
­
040700POOL.
The
test
sample
was
a
mixture
of
the
two
PS149B1
proteins,
14
kDa
and
44
kDa.
These
were
formulated
in
a
1:
4.6
ratio
of
54
%
pure
14
kDa
and
37
%
pure
44
kDa
proteins.
The
intention
was
to
deliver
a
dose
that
contained
482
mg/
kg
of
pure
14
kDa
and
1520
mg/
kg
of
pure
44
kDa
protein;
this
1:
3
ratio
provided
an
equimolar
mixture
of
the
proteins.
A
single
dose
gavage
containing
a
20
%
mixture
of
proteins
in
0.5
%
aqueous
methylcellulose
was
administered.

Reference
number
­
PS149B1
14
kDa
­
TSN102172,
44
kDa
­
TSN
102171.

Physical
properties
­
Off­
white
solid.

Test
animals
­
CD­
1
mice
(
Charles
River
Laboratories,
Portage,
MI)
weighing
23
to
35
g
at
the
start
of
the
study
were
dosed
on
July
25,
2000
or
August
1,
2000.
Relative
humidity
was
maintained
at
40
to
72
%
and
temperature
22
+
3

C
in
the
animal
room.
A
12
hour
photoperiod
was
used
and
room
air
was
exchanged
approximately
12
to
15
times
per
hour.
Water
lines
were
automatically
bled
every
six
hours.

Purina
Certified
Rodent
Lab
Diet
#
5002
(
Purina
Mills,
Inc.,
St.
Louis,
MO)
and
municipal
water
were
supplied
ad
libitum.

Experimental
design
­
The
test
substance
was
administered
to
five
female
and
five
male
mice
(
5000
mg/
kg
body
weight)
in
a
1:
4.6
mixture
of
the
two
PS149B1
proteins,
14
kDa
and
44
kDa.
A
single
dose
gavage
(
25
mL/
kg)
delivered
as
a
20
%
mixture
in
0.5
%
methycellulose.
Mice
were
fasted
one
hour
prior
to
treatment.
Detailed
clinical
observations
were
made
prior
to
the
dosing
for
comparison
to
study
observations.
Animals
were
observed
at
least
twice
on
the
day
of
dosing
and
once
daily
thereafter
a
detailed
clinical
observation
was
performed.
each
animal
was
weighed
prestudy,
on
the
day
of
dosing
and
at
days
2,
8,
and
15.
Gross
necropsy
was
performed
on
all
animals.

Pathology
­
All
mice
surviving
the
study
were
euthanized
and
examined
by
gross
necropsy.
Eyes
were
examined
in
situ
using
a
glass
microscope
slide
applied
to
the
corneum.
External
surfaces,
body
orifices,
nasal,
cranial,
oral,
thoracic
and
abdominal
cavities
were
all
opened
and
the
viscera
examined
in
situ
and
after
dissection.

Statistics
­
means
and
standard
deviations
were
calculated
relative
to
body
weight
measurements.

RESULTS
All
animals
survived
the
two
week
study.
One
female
mouse
exhibited
protruding
or
bulging
eyes
on
days
6
and
7,
but
this
resolved
thereafter.
This
observation
was
not
attributed
to
the
treatment
as
it
was
an
isolated
observation
(
i.
e.,
no
other
animals
exhibited
this).
No
other
clinical
signs
were
noted
for
any
animals
during
the
study.
31
An
initial
weight
loss
was
observed
in
two
mice
at
test
days
1
and
2,
but
both
gained
weight
for
the
remainder
of
the
study.
Given
the
timing
of
this
observation,
the
dosage
administration
may
likely
be
responsible
for
the
initial
weight
loss
response,
particularly
with
the
inclusion
of
the
methycellulose
as
a
carrier
during
dosing.
All
other
animals
gained
weight
throughout
the
study.

No
gross
treatment
related
observations
were
recorded
during
the
study
as
represented
by
gross
pathologic
observations.

An
acute
oral
LD50
was
calculated
for
this
study
based
upon
a
dosage
of
a
1:
4.6
ratio
mixture
of
PS149B1
proteins
at
greater
than
5000
mg/
kg
and
greater
than
2000
mg/
kg
for
an
equimolar
mixture
(
1:
3)
of
the
pure
proteins.