Document ID: EPA-HQ-OPP-2003-0024-0024
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2003-06-03T04:00Z

Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
1
of
41
Data
Requirement:
:
EPA
DP
Barcode
D288775
EPA
MRID
458677­
02
EPA
Guideline
70­
1(
Special
Study)

Test
material:
Purity:
not
reported
Common
name
Atrazine
Chemical
name:
IUPAC
CAS
name
6­
chloro­
N­
ethyl­
N'­(
1­
methylethyl)­
1,3,5­
triazine­
2,4­
diamine
CAS
No.
1912­
24­
9
Synonyms
EPA
PC
Code:
80803
Primary
Reviewer:
Thomas
M.
Steeger,
Ph.
D.,
Senior
Biologist
Date:
March
27,
2003
Environmental
Fate
and
Effects
Division,
ERB
4,
U.
S.
Environmental
Protection
Agency
Secondary
Reviewer(
s):
Joseph
E.
Tietge,
M.
S.,
Research
Aquatic
Biologist
Date:
Mid­
Continent
Ecology
Division,
National
Health
and
Environmental
Effects
Research
Laboratory
(
Duluth),
U.
S.
Environmental
Protection
Agency
Stephanie
Irene,
Ph.
D.,
Senior
Advisor
Date:
Environmental
Fate
and
Effects
Division,
ERB
3,
U.
S.
Environmental
Protection
Agency
Mary
J.
Frankenberry,
Senior
Statistician
Date:
Environmental
Fate
and
Effects
Division,
ERB
3,
U.
S.
Environmental
Protection
Agency
EPA
PC
Code
080803
Date
Evaluation
Completed:
06/
01/
2003
CITATION:
Jones,
P.
D.,
M.
B.
Murphy,
M.
Hecker,
J.
P.
Giesy.
2003.
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
Aquatic
Toxicology
Laboratory,
Michigan
State
University,
218C
National
Food
Safety
and
Toxicology
Center,
E.
Lansing,
MI.
Sponsor:
Syngenta
Crop
Protection,
Inc.
Laboratory
Study
ID:
ECORISK
Number
MSU­
02.
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
2
of
41
EXECUTIVE
SUMMARY:

This
represents
a
summary
for
Phase
I
of
a
three­
year
study
where
primarily
the
green
frog
(
Rana
clamitans)
but
also
the
leopard
frog
(
Rana
pipiens)
and
the
bullfrog
(
Rana
catesbeiana)
were
collected
from
three
reference
ponds
and
six
atrazine­
exposed
ponds.
Both
juvenile
(
372)
and
adult
(
340)
green
frogs
were
collected
and
examined
for
gross
gonadal
abnormalities
and
blood
plasma
estradiol
and
testosterone
levels.
A
total
of
four
mixed
or
unknown
sex
animals
were
identified
in
all
of
the
frogs
collected.
Hormone
levels
exhibited
considerable
variability
among
locations
and
individuals.
No
relationship
between
plasma
hormone
levels
and
atrazine
exposure
could
be
determined.

While
this
represents
an
interim
report
(
Phase
1)
of
a
three­
year
study,
the
results
suggest
that
Rana
clamitans
are
not
markedly
impacted
by
atrazine
exposure
in
terms
of
gonadal
deformities.
However,
reference
sites
contained
atrazine
so
the
ability
of
this
field
study
to
discriminate
effects
may
be
limited.
Additionally,
sex
steroid
levels
exhibited
considerable
variability,
and
the
study
was
not
able
to
distinguish
meaningful
differences
between
"
exposed"
and
"
reference"
sites.
Plasma
steroid
levels
are
of
questionable
value
because
data
were
collected
over
a
time
period
of
four
months
for
adult
frogs,
and
coefficients
of
variability
ranged
as
high
as
10,628%.
There
were
no
gender­
specific
estradiol
levels.
Exposed
males
contained
roughly
similar
(
0.91)
amounts
of
estradiol
as
females,
while
in
reference
sites
males
exhibited
roughly
4.9
times
the
plasma
estradiol
concentration
than
females.

Because
of
the
variability
associated
with
plasma
steroid
hormone
levels
measured
in
this
study,
it
is
unlikely
that
the
study
will
be
able
to
differentiate
any
treatment­
related
effects,
especially
since
the
reference
sites
may
have
contained
low
levels
of
atrazine.
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
3
of
41
I.
MATERIALS
AND
METHODS
GUIDELINE
FOLLOWED:
Nonguideline
Study
COMPLIANCE:
Not
conducted
under
full
GLP;
however,
most
practices
as
defined
by
40
CFR
Part
160,
August
19,
1989
were
established
for
this
study,
including
but
not
limited
to:

°
Written,
authorized
protocol
°
Written,
authorized
Standard
Operating
Procedures
for
all
key
procedures.
°
Organization
and
Personnel
were
sufficient
in
terms
of
number,
education,
training
and
experience.
°
Facilities
were
of
suitable
size
and
construction
°
Equipment
used
was
of
appropriate
design
and
adequate
capacity.
°
Independent
QA
Inspection
were
conducted.
°
Final
Report
was
written
°
Raw
data,
documentation,
records,
protocols,
and
final
report
were
archived.

A.
MATERIALS:

1.
Test
Material
Atrazine
Description:
Not
reported
Lot
No./
Batch
No.
:
Not
reported
Purity:
NA
Stability
of
compound
under
test
conditions:
Not
reported
Storage
conditions
of
test
chemicals:
Not
reported
2.
Test
organism:

Species:
Green
Frogs
(
Rana
clamitans)
Age
at
test
initiation:
Juveniles
and
adults
Weight
at
study
initiation:
(
mean
and
range)
Not
reported
Length
at
study
initiation:
(
mean
and
range)
Not
reported
Source:
Field­
collected
at
nine
(
3
reference
and
6
atrazine­
exposed)
sites
located
in
Livingston,
Ingham,
Barry
and
Lapeer
counties,
Michigan
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
4
of
41
B.
STUDY
DESIGN:

Objective:
To
assess
the
effects
of
atrazine
on
kidney
and
gonad
histology
and
plasma
steroid
hormone
concentrations
and
gonadal
aromatase
activity
of
green
frogs
(
R.
clamitans)
and
other
incidental
ranid
species
collected
from
various
field
sites
in
Michigan.

1.
Experimental
Conditions
Experimental
sites
(
KZ02,
KZ03,
KZ07,
LPR02,
LPR03
and
HW01)
were
based
on
proximity
to
agriculture
where
atrazine
was
used
and
where
the
genus
Rana
were
located.
Reference
sites
(
HG01,
HL01
and
LPR08)
were
selected
in
non­
agricultural
areas.

During
each
sampling,
water
chemistry
(
conductivity,
DO,
pH
and
temperature
at
each
of
4
points
on
water's
surface)
samples
were
collected,
and
on
the
first
visit
a
sediment
(
5­
cm
grab)
sample
was
collected.
All
samples
were
analyzed
for
atrazine
using
Envirogard
triazine
ELISA
kit
(
LOD
=
0.025

g/
L);
sediment
samples
will
be
extracted
and
then
analyzed
later
using
ELISA.
Results
of
ELISA
will
be
verified
using
GC­
MS.

Sampling
(
from
8
PM
to
midnight)
was
conducted
three
times
throughout
the
Summer
of
2002.
Tadpoles
were
collected
in
un­
baited
minnow
traps
in
the
late
spring
and
early
summer;
juvenile
frogs
were
sampled
in
mid­
summer
(
June
and
July);
adult
frogs
were
sampled
at
the
end
of
summer
(
September).
Frogs
were
sampled
using
fish
nets
and
were
held
in
buckets
for
less
than
two
hours
to
minimize
sampling
stress
and
affects
on
plasma
sex
steroid
concentrations.

Frogs
were
euthanized
by
immersion
in
MS­
222,
then
weighed
and
measured
(
SVL)
before
blood
collection
via
cardiac
puncture;
blood
collection
was
completed
within
an
approximate
four­
hour
window
of
time.

Frogs
were
dissected
and
examined
under
a
microscope
for
gonad
morphology.
One
gonad
was
collected
from
adult
and
juvenile
frogs,
and
snap
frozen
for
use
in
aromatase
assay.
The
other
gonad
was
left
in
the
carcass
and
fixed
in
Bouin's
solution.
Gonadal
somatic
index
(
partial
GSI)
was
calculated
based
on
the
weight
of
single
gonad.

Plasma
samples
were
extracted
with
diethyl
ether.
The
level
of
quantitation
(
LOQ)
for
testosterone
and
17­
 
estradiol
was
0.78
­
800
pg/
well.

II.
RESULTS
and
DISCUSSION:
[
All
results
discussed
in
this
section
and
the
next
are
those
reported
by
the
study
authors.
Although
supplemental
data
are
typically
used
in
a
qualitative
manner
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
5
of
41
only,
EFED
verified
spreadsheet
data
and
ran
basic
statistical
analyses
on
the
major
study
parameters.
See
attached
appendix.
If
results
differed
in
any
substantive
way,
the
difference
was
reported
in
the
text
below.]

Quantifiable
levels
of
atrazine
were
detected
in
all
(
exposed
and
reference)
of
the
ponds
sampled.
Atrazine
concentrations
in
exposed
ponds
ranged
from
0.025
to
250

g/
L.
Reference
pond
water
atrazine
concentrations
ranged
from
0.015
to
0.093

g/
L.
No
data
were
reported
on
other
triazine
herbicides,
degradates
or
other
pesticide
residues.
Although
the
study
notes
that
maximum
concentrations
did
not
appear
to
persist
for
very
long
at
the
sites,
with
measurements
tending
to
drop
off
in
the
next
water
sample,
intervals
between
sampling
make
it
difficult
to
support
this
observation.

Table
1.
Atrazine
water
concentrations
in
both
exposed
and
reference
collection
sites
during
spring
(
May),
early
summer
(
June
and
July),
mid­
summer
(
August)
and
late
summer
(
September)
samples.

Site
Atrazine
Water
Concentrations
(

g/
L)

Spring
Early­
Summer
Mid­
Summer
Late­
Summer
KZ02
(
Exposed
1)
0.52
0.094
0.025
0.030
KZ03
(
Exposed
2)
1.6
1.8
0.71
0.11
KZ07
(
Exposed
3)
0.031
0.48
0.17
0.11
HW01
(
Exposed
4)
0.12
0.36
0.24
0.24
LPR02
(
Exposed
5)
0.83
6.5
x
101
2.9
0.078
LPR03
(
Exposed
6)
2.5
x
102
0.69
0.092
1.9
HL01
(
Reference
1)
0.058
0.069
0.040
0.015
HG01
(
Reference
2)
0.026
0.093
0.069
0.067
LPR08
(
Reference
3)
0.085
0.062
0.033
0.036
Of
the
372
juvenile
and
340
adult
green
frogs
collected,
four
(
0.6%)
had
mixed
or
unknown
sex.
It
is
assumed
that
this
is
based
on
dissecting
scope
observations
because
it
states
that
animals
will
be
subject
to
histopathology
in
upcoming
months.

GSI
varied
considerably
between
sampling
sites.
Female
GSI
was
greatest
at
HL01
reference
site
and
at
the
HW01
and
LPR02
exposed
sites.

Within­
site
and
between­
site
variability
in
testosterone
(
T)
levels
was
high
(
Tables
2
and
3).
Testosterone
was
highest
at
LPR08
reference
site
and
KZ02
experimental
site
in
both
males
and
females
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
6
of
41
Estradiol
(
E
2)
concentrations
in
both
male
and
females
was
greatest
at
KZ02
exposed
site
and
LPR08
reference
site
(
Tables
2
and
3).

The
ratio
of
plasma
estradiol
to
testosterone
levels
(
E/
T)
in
females
averaged
1.7
±
1.6
at
atrazine­
exposed
sites
compared
to
1.6
±
1.0
at
reference
sites
(
Table
4).
For
males,
E/
T
ratios
averaged
1.28
±
1.7
and
0.95
±
0.6
for
atrazine­
exposed
and
reference
sites,
respectively.

Table
2.
Mean
plasma
testosterone
and
estradiol
in
female
green
frogs
(
Rana
clamitans)
collected
in
reference
and
atrazine­
exposed
sites
in
Michigan.
Standard
deviations
(
Std.
Dev.)
and
coefficients
of
variability
(
CV
=
[
std.
dev.
÷
mean]
x
100).

Site
Female
Rana
clamitans
Mean
Testosterone
pg/
mL
Std.
Dev.
CV
Mean
Estradiol
pg/
mL
Std.
Dev.
CV
Exposed
1
(
KZ02)
3697
641
17%
16407
5718
35%

Exposed
2
(
KZ03)
198
2101
1061%
495
4573
924%

Exposed
3
(
KZ07)
249
1684
676%
289
478
165%

Exposed
4
(
HW01)
640
534
83%
25
226
904%

Exposed
5
(
LPR02)
341
843
247%
68
826
1215%

Exposed
6
(
LPR03)
209
188
90%
321
467
145%

Reference
1
(
HL01)
180
646
359%
134
183
137%

Reference
2
(
HG01)
134
98
73%
51
172
337%

Reference
3
(
LPR08)
3681
3452
94%
5253
4430
84%

Table
3.
Mean
plasma
testosterone
and
estradiol
in
male
green
frogs
(
Rana
clamitans)
collected
in
reference
and
atrazine­
exposed
sites
in
Michigan.
Standard
deviations
(
Std.
Dev.)
and
coefficients
of
variability
(
CV
=
[
std.
dev.
÷
mean]
x
100).

Site
Male
Rana
clamitans
Mean
Testosterone
pg/
mL
Std.
Dev.
CV
Mean
Estradiol
pg/
mL
Std.
Dev.
CV
Exposed
1
(
KZ02)
3300
8574
260%
14308
7668
54%

Exposed
2
(
KZ03)
118
888
753%
238
7678
3226%

Exposed
3
(
KZ07)
129
3555
2756%
20
1199
5995%
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
7
of
41
Exposed
4
(
HW01)
89
­­
­­
26
­­
­­

Exposed
5
(
LPR02)
168
173
103%
2.7
133
4926%

Exposed
6
(
LPR03)
826
­­
­­
732
­­
­­

Reference
1
(
HL01)
149
610
409%
36
3826
10628%

Reference
2
(
HG01)
189
1733
917%
234
2956
1263%

Reference
3
(
LPR08)
3183
4171
131%
4314
3568
83%

Table
4.
Female
and
male
plasma
estradiol
to
testosterone
(
E/
T)
ratios
for
green
frogs
(
Rana
clamitans)
collected
from
atrazine­
exposed
and
reference
sites.

Site
Female
E/
T
Ratio
Male
E/
T
Ratio
Exposed
1
(
KZ02)
4.4
4.34
Exposed
2
(
KZ03)
2.5
2.02
Exposed
3
(
KZ07)
1.2
0.16
Exposed
4
(
HW01)
0.04
0.29
Exposed
5
(
LPR02)
0.20
0.02
Exposed
6
(
LPR03)
1.54
0.89
Reference
1
(
HL01)
0.74
0.24
Reference
2
(
HG01)
0.38
1.24
Reference
3
(
LPR08)
1.43
1.36
REVIEWER'S
COMMENTS:

Because
of
the
large
number
of
sampling
sites
and
the
logistics
of
collecting
and
processing
samples
at
night,
it
is
understandable
why
sampling
had
to
be
conducted
over
a
protracted
period
of
time.
However,
while
many
of
the
experimental
sites
were
sampled
between
7/
23/
02
and
7/
29/
02,
sampling
of
reference
sites
extended
from
7/
31/
02
through
8/
13/
02.
Only
one
exposed
site
(
LPR02)
was
sampled
as
late
as
8/
13/
02.
When
adult
frogs
were
collected
in
September,
reference
sites
were
sampled
relatively
early
(
9/
6
­
9/
16),
while
experimental
areas
were
sampled
later
(
9/
13
­
9/
30).
It's
not
clear
when
each
of
the
samples
was
collected;
there
are
sampling
dates
reported
in
the
appendix,
but
these
are
not
consistent.
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
8
of
41
.
The
major
hypothesis
that
was
being
evaluated
in
this
study
is
that
atrazine
exposure
results
in
abnormal
kidney
and
gonad
development
and
that
atrazine
causes
changes
in
plasma
estradiol
and
testosterone
levels.
Because
of
the
design
of
the
experiment
and
the
uncontrolled
environmental
factors
in
a
field
study,
this
hypothesis
could
not
be
tested.
Furthermore,
presumptive
effects
on
gonadal
development
must
occur
at
the
appropriate
time
in
development..
In
this
study,
atrazine
measurements
were
taken
throughout
the
summer,
but
they
were
apparently
not
taken
during
the
larval
period.
As
a
result,
it
is
uncertain
whether
or
not
exposure
occurred
at
the
appropriate
time
to
elicit
a
response.

Because
there
is
considerable
variability
in
the
plasma
steroid
levels,
it
is
difficult
to
determine
potential
treatment
effects
based
on
these
measurements.
Plasma
estradiol
levels
in
atrazine­
exposed
females
ranged
from
25
to
16,407
pg/
mL
with
coefficients
of
variability
(
CV)
ranging
from
35%
to
1,215%;
at
reference
sites
female
plasma
estradiol
ranged
from
51
to
5,253
pg/
mL
with
CV's
ranging
from
84
to
337%.
Testosterone
levels
in
females
were
equally
variable
with
coefficients
of
variability
ranging
between
17
and
1,061%;
mean
plasma
testosterone
levels
in
females
from
both
atrazine­
exposed
and
reference
sites
ranged
roughly
from
134
to
3,697
pg/
mL.
For
males,
plasma
testosterone
levels
in
atrazine­
exposed
animals
ranged
from
89
to
3,300
pg/
mL
(
CV
range
103
­
2,756%),
while
males
from
reference
sites
ranged
from
149
to
3,183
pg/
mL
(
CV
range
131
­
917%).
Plasma
estradiol
levels
in
males
at
atrazine­
exposed
sites
ranged
from
2.7
to
14,308
pg/
mL
(
CV
range
54
­
5,995%),
while
reference
site
males
ranged
from
36
­
4314
pg/
mL
(
CV
range
83
­
10,628%).
Iinteresting
enough
the
highest
plasma
estradiol
ranges
were
measured
in
males
collected
from
atrazineexposed
sites.
Unfortunately,
the
highest
variability
(
as
reflected
by
CVs
ranging
as
high
as
10,628%)
was
observed
in
reference
sites
for
male
estradiol
levels.
With
such
high
levels
of
variability
and
with
the
presence
of
atrazine
in
the
reference
sites,
the
potential
to
differentiate
treatment
effects
would
be
remote,
unless
extremely
large
sample
sizes
were
used.
Alternatively
the
researchers
could
use
a
shorter
time
period
for
collecting
and
analyzing
samples
to
limit
the
likelihood
that
frogs
would
be
at
different
stages
of
their
breeding
cycle.

Gonadal
anomalies
were
only
detected
in
0.6%
of
the
total
juvenile
and
adult
frogs
sampled;
however,
it
is
unclear
whether
histopathology
will
significantly
alter
that
percentage.

The
median
ratio
of
estradiol
to
testosterone
exceeded
1.0
(
range:
1.9
­
8.6)
in
males
collected
from
reference
sites;
however,
at
half
of
the
exposed
sites
the
median
ratio
was
less
than
1.0
(
range
0.26
­
0.89).
The
remaining
exposed
sites
(
3)
had
median
ratios
ranging
from
4.9
to
6.8.

Reference
sites
all
proved
to
contain
measurable
levels
of
atrazine
and
were
in
some
cases
higher
than
exposed
site
values.
No
data
were
reported
on
other
triazines
and/
or
degradates
nor
were
data
provided
on
what
other
chemicals
may
have
been
present
in
the
exposed
and
reference
study
sites.
Although
water
quality
data
were
collected,
no
data
were
provided
for
analysis.

While
this
represents
an
interim
report
(
Phase
1)
of
a
three­
year
study,
the
results
suggest
that
Rana
clamitans
were
not
markedly
impacted
by
atrazine
exposure
in
terms
of
gonadal
deformities.
However,
reference
sites
contained
atrazine
so
the
ability
of
this
field
study
to
discriminate
effects
may
be
limited.
Additionally,
sex
steroid
levels
exhibited
considerable
variability,
and
the
study
was
not
able
to
distinguish
meaningful
differences
between
"
exposed"
and
"
reference"
sites.
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
9
of
41
The
summary
tables
for
plasma
steroid
levels
are
of
little
value
because
they
summarize
data
collected
over
4
months
in
adult
frogs.
There
were
no
gender­
specific
steroid
hormone
levels,
both
males
and
females
exhibited
roughly
similar
average
ratios
from
atrazine
exposed
sites,
i.
e.,
1.7
versus
1.3,
and
from
reference
sites,
i.
e.,
0.85
and
0.95,
suggesting
that
this
study
couldn't
distinguish
plasma
steroid
differences
between
males
and
females
or
between
reference
and
exposure
sites
if
such
differences
existed.

Although
the
study
was
not
conducted
under
full
GLP,
the
report
notes
that
most
practices
were
included,
one
of
which
involved
the
writing
of
a
final
report.
The
current
study
report
does
not
constitute
a
final
report
and
therefore
a
Final
Report
was
not
written.

CONCLUSIONS:

Although
this
study
provides
preliminary
data
on
the
incidence
of
abnormal
gonads
and
the
levels
of
plasma
steroid
hormone
in
green
frogs
collected
from
both
reference
and
atrazine­
exposed
sites,
there
is
no
conclusive
evidence
that
frogs
from
the
two
sites
are
statistically
different
relative
to
these
measurement
endpoints.
However,
the
gonadal
data
are
based
on
gross
morphology
and
the
incidence
of
abnormalities
may
change
once
histology
is
completed.
Additionally,
given
the
high
level
of
variability
in
steroid
hormone
levels,
it
is
unlikely
that
this
study
could
have
differentiated
between
males
and
females
or
between
reference
and
atrazine­
exposed
sites
had
a
difference
existed.
With
coefficients
of
variability
approaching
11,000%,
the
number
of
animals
required
to
detect
a
specified
difference
with
any
reasonable
level
of
confidence
would
be
high
given
the
study's
current
design.
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
10
of
41
AVERAGE
WEIGHT
OF
FROGS
COLLECTED
BY
AREA
AND
SEX
279
Obs
AREA
SEX
_
TYPE_
_
FREQ_
MEAN
STD
CV
1
HG01
0
1
.
.
.
2
HG01
F
0
4
44.9828
11.6195
25.8310
3
HG01
M
0
24
40.7518
11.0699
27.1643
4
HL01
F
0
7
45.7543
4.4464
9.7179
5
HL01
M
0
18
40.1478
7.7085
19.2003
6
HL02
F
0
2
73.0950
13.7674
18.8349
7
HW01
F
0
4
42.0025
23.7297
56.4958
8
HW01
M
0
1
18.4600
.
.
9
KZ03
F
0
13
36.5923
14.0417
38.3734
10
KZ03
M
0
9
30.5756
10.3927
33.9903
11
KZ07
F
0
22
32.2627
6.3247
19.6037
12
KZ07
M
0
28
29.9182
6.1725
20.6312
13
KZ07
Unk.
0
1
30.7600
.
.
14
LPR0
F
0
23
31.5283
15.8872
50.3904
15
LPR0
M
0
24
28.4154
10.1072
35.5696
NONPARAMETRIC
COMPARISON
OF
BODY
WEIGHT
ACROSS
SAMPLING
AREAS
BY
SEX
280
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
F
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
NPAR1WAY
Procedure
Wilcoxon
Scores
(
Rank
Sums)
for
Variable
WEIGHT
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
4
222.0
152.0
42.410690
55.500000
HL01
7
423.0
266.0
54.905980
60.428571
HL02
2
146.0
76.0
30.408332
73.000000
HW01
4
166.0
152.0
42.410690
41.500000
KZ03
13
481.0
494.0
71.446950
37.000000
KZ07
22
721.0
836.0
85.934083
32.772727
LPR0
23
691.0
874.0
87.032561
30.043478
Kruskal­
Wallis
Test
Chi­
Square
19.6115
DF
6
Pr
>
Chi­
Square
0.0032
NONPARAMETRIC
COMPARISON
OF
BODY
WEIGHT
ACROSS
SAMPLING
AREAS
BY
SEX
281
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
F
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
NPAR1WAY
Procedure
Median
Scores
(
Number
of
Points
Above
Median)
for
Variable
WEIGHT
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
4
4.0
1.973333
0.979433
1.000000
HL01
7
7.0
3.453333
1.267999
1.000000
HL02
2
2.0
0.986667
0.702250
1.000000
HW01
4
2.0
1.973333
0.979433
0.500000
KZ03
13
6.0
6.413333
1.649997
0.461538
KZ07
22
8.0
10.853333
1.984563
0.363636
LPR0
23
8.0
11.346667
2.009931
0.347826
Median
One­
Way
Analysis
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
11
of
41
Chi­
Square
16.6089
DF
6
Pr
>
Chi­
Square
0.0108
NONPARAMETRIC
COMPARISON
OF
BODY
WEIGHT
ACROSS
SAMPLING
AREAS
BY
SEX
282
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
M
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
NPAR1WAY
Procedure
Wilcoxon
Scores
(
Rank
Sums)
for
Variable
WEIGHT
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
24
1717.0
1260.00
129.614814
71.541667
HL01
18
1299.0
945.00
116.382988
72.166667
HW01
1
8.0
52.50
30.020826
8.000000
KZ03
9
385.0
472.50
86.494219
42.777778
KZ07
28
1160.0
1470.00
136.455121
41.428571
LPR0
24
891.0
1260.00
129.614814
37.125000
Kruskal­
Wallis
Test
Chi­
Square
30.3302
DF
5
Pr
>
Chi­
Square
<.
0001
Median
Scores
(
Number
of
Points
Above
Median)
for
Variable
WEIGHT
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
24
18.0
12.00
2.158748
0.750000
HL01
18
15.0
9.00
1.938371
0.833333
HW01
1
0.0
0.50
0.500000
0.000000
KZ03
9
3.0
4.50
1.440570
0.333333
KZ07
28
9.0
14.00
2.272674
0.321429
LPR0
24
7.0
12.00
2.158748
0.291667
Median
One­
Way
Analysis
Chi­
Square
23.5098
DF
5
Pr
>
Chi­
Square
0.0003
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
12
of
41
AVERAGE
LENGTH
OF
FROGS
COLLECTED
BY
AREA
AND
SEX
284
Obs
AREA
SEX
_
TYPE_
_
FREQ_
MEAN
STD
CV
1
HG01
0
1
.
.
.
2
HG01
F
0
4
7.77475
0.62670
8.0608
3
HG01
M
0
24
7.28233
1.61161
22.1304
4
HL01
F
0
7
7.69100
0.34031
4.4247
5
HL01
M
0
18
7.50867
0.51030
6.7962
6
HL02
F
0
2
8.94050
0.84782
9.4829
7
HW01
F
0
4
7.38650
1.13260
15.3334
8
HW01
M
0
1
5.87600
.
.
9
KZ03
F
0
13
7.32492
1.02200
13.9523
10
KZ03
M
0
9
6.96600
0.75002
10.7669
11
KZ07
F
0
22
6.98036
0.48803
6.9915
12
KZ07
M
0
28
6.86843
0.51202
7.4547
13
KZ07
Unk.
0
1
7.08900
.
.
14
LPR0
F
0
23
6.79417
1.23124
18.1220
15
LPR0
M
0
24
6.73617
0.81422
12.0874
NONPARAMETRIC
COMPARISON
OF
BODY
LENGHT
(
SVL)
ACROSS
SAMPLING
AREAS
BY
SEX
285
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
F
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
NPAR1WAY
Procedure
Wilcoxon
Scores
(
Rank
Sums)
for
Variable
SVL
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
4
221.0
152.0
42.410690
55.250000
HL01
7
393.0
266.0
54.905980
56.142857
HL02
2
142.0
76.0
30.408332
71.000000
HW01
4
178.0
152.0
42.410690
44.500000
KZ03
13
483.0
494.0
71.446950
37.153846
KZ07
22
712.0
836.0
85.934083
32.363636
LPR0
23
721.0
874.0
87.032561
31.347826
Kruskal­
Wallis
Test
Chi­
Square
15.9314
DF
6
Pr
>
Chi­
Square
0.0141
Median
Scores
(
Number
of
Points
Above
Median)
for
Variable
SVL
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
4
4.0
1.973333
0.979433
1.000000
HL01
7
7.0
3.453333
1.267999
1.000000
HL02
2
2.0
0.986667
0.702250
1.000000
HW01
4
3.0
1.973333
0.979433
0.750000
KZ03
13
3.0
6.413333
1.649997
0.230769
KZ07
22
9.0
10.853333
1.984563
0.409091
LPR0
23
9.0
11.346667
2.009931
0.391304
Median
One­
Way
Analysis
Chi­
Square
19.3126
DF
6
Pr
>
Chi­
Square
0.0037
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
13
of
41
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
14
of
41
NONPARAMETRIC
COMPARISON
OF
BODY
LENGHT
(
SVL)
ACROSS
SAMPLING
AREAS
BY
SEX
287
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
M
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
NPAR1WAY
Procedure
Wilcoxon
Scores
(
Rank
Sums)
for
Variable
SVL
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
24
1642.00
1260.00
129.614123
68.416667
HL01
18
1251.00
945.00
116.382368
69.500000
HW01
1
9.00
52.50
30.020666
9.000000
KZ03
9
416.00
472.50
86.493758
46.222222
KZ07
28
1176.50
1470.00
136.454393
42.017857
LPR0
24
965.50
1260.00
129.614123
40.229167
Average
scores
were
used
for
ties.

Kruskal­
Wallis
Test
Chi­
Square
22.2193
DF
5
Pr
>
Chi­
Square
0.0005
Median
Scores
(
Number
of
Points
Above
Median)
for
Variable
SVL
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
24
17.0
12.00
2.158748
0.708333
HL01
18
12.0
9.00
1.938371
0.666667
HW01
1
0.0
0.50
0.500000
0.000000
KZ03
9
4.0
4.50
1.440570
0.444444
KZ07
28
11.0
14.00
2.272674
0.392857
LPR0
24
8.0
12.00
2.158748
0.333333
Average
scores
were
used
for
ties.

Median
One­
Way
Analysis
Chi­
Square
11.1222
DF
5
Pr
>
Chi­
Square
0.0490
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
15
of
41
AVERAGE
GONAD
WEIGHT
OF
FROGS
COLLECTED
BY
AREA
AND
SEX
289
Obs
AREA
SEX
_
TYPE_
_
FREQ_
MEAN
STD
CV
1
HG01
0
1
.
.
.
2
HG01
F
0
4
0.53175
0.28141
52.922
3
HG01
M
0
24
0.08608
0.02893
33.607
4
HL01
F
0
7
0.79743
0.27233
34.151
5
HL01
M
0
18
0.07022
0.02304
32.815
6
HL02
F
0
2
4.20200
2.90055
69.028
7
HW01
F
0
4
0.90725
1.05756
116.568
8
HW01
M
0
1
0.02600
.
.
9
KZ03
F
0
13
0.43562
0.48035
110.269
10
KZ03
M
0
9
0.06600
0.02666
40.401
11
KZ07
F
0
22
0.27882
0.14849
53.257
12
KZ07
M
0
28
0.06400
0.01933
30.205
13
KZ07
Unk.
0
1
.
.
.
14
LPR0
F
0
23
0.35174
0.40409
114.884
15
LPR0
M
0
24
0.07358
0.08499
115.502
NONPARAMETRIC
COMPARISON
OF
GONAD
WEIGHT
ACROSS
SAMPLING
AREAS
BY
SEX
290
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
F
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
NPAR1WAY
Procedure
Wilcoxon
Scores
(
Rank
Sums)
for
Variable
GONAD
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
4
189.00
152.0
42.409786
47.250000
HL01
7
433.00
266.0
54.904809
61.857143
HL02
2
148.00
76.0
30.407683
74.000000
HW01
4
188.00
152.0
42.409786
47.000000
KZ03
13
482.50
494.0
71.445426
37.115385
KZ07
22
715.50
836.0
85.932250
32.522727
LPR0
23
694.00
874.0
87.030704
30.173913
Average
scores
were
used
for
ties.

Kruskal­
Wallis
Test
Chi­
Square
19.6246
DF
6
Pr
>
Chi­
Square
0.0032
Median
Scores
(
Number
of
Points
Above
Median)
for
Variable
GONAD
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
4
3.0
1.973333
0.979433
0.750000
HL01
7
7.0
3.453333
1.267999
1.000000
HL02
2
2.0
0.986667
0.702250
1.000000
HW01
4
2.0
1.973333
0.979433
0.500000
KZ03
13
5.0
6.413333
1.649997
0.384615
KZ07
22
9.0
10.853333
1.984563
0.409091
LPR0
23
9.0
11.346667
2.009931
0.391304
Average
scores
were
used
for
ties.
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
16
of
41
Median
One­
Way
Analysis
Chi­
Square
12.3288
DF
6
Pr
>
Chi­
Square
0.0550
NONPARAMETRIC
COMPARISON
OF
GONAD
WEIGHT
ACROSS
SAMPLING
AREAS
BY
SEX
292
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
M
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
NPAR1WAY
Procedure
Wilcoxon
Scores
(
Rank
Sums)
for
Variable
GONAD
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
24
1642.00
1248.0
128.153570
68.416667
HL01
18
983.50
936.0
115.121717
54.638889
HW01
1
6.00
52.0
29.724299
6.000000
KZ03
9
417.00
468.0
85.604525
46.333333
KZ07
27
1335.00
1404.0
133.321509
49.444444
LPR0
24
972.50
1248.0
128.153570
40.520833
Average
scores
were
used
for
ties.

Kruskal­
Wallis
Test
Chi­
Square
13.8281
DF
5
Pr
>
Chi­
Square
0.0167
Median
Scores
(
Number
of
Points
Above
Median)
for
Variable
GONAD
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
24
17.0
11.883495
2.155602
0.708333
HL01
18
10.0
8.912621
1.936400
0.555556
HW01
1
0.0
0.495146
0.499976
0.000000
KZ03
9
3.0
4.456311
1.439908
0.333333
KZ07
27
13.0
13.368932
2.242529
0.481481
LPR0
24
8.0
11.883495
2.155602
0.333333
Average
scores
were
used
for
ties.

Median
One­
Way
Analysis
Chi­
Square
8.9955
DF
5
Pr
>
Chi­
Square
0.1092
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
17
of
41
AVERAGE
GONADOSOMATIC
INDEX
(
GSI)
OF
FROGS
COLLECTED
BY
AREA
AND
SEX
294
Obs
AREA
SEX
_
TYPE_
_
FREQ_
MEAN
STD
CV
1
HG01
0
1
.
.
.
2
HG01
F
0
4
0.011275
0.004655
41.286
3
HG01
M
0
24
0.002137
0.000476
22.280
4
HL01
F
0
7
0.017357
0.005367
30.924
5
HL01
M
0
18
0.001717
0.000333
19.397
6
HL02
F
0
2
0.054700
0.029416
53.776
7
HW01
F
0
4
0.016925
0.011787
69.640
8
HW01
M
0
1
0.001400
.
.
9
KZ03
F
0
13
0.009985
0.006135
61.443
10
KZ03
M
0
9
0.002156
0.000343
15.921
11
KZ07
F
0
22
0.008295
0.003194
38.508
12
KZ07
M
0
28
0.002178
0.000571
26.234
13
KZ07
Unk.
0
1
.
.
.
14
LPR0
F
0
23
0.009487
0.007301
76.955
15
LPR0
M
0
24
0.002421
0.002454
101.367
NONPARAMETRIC
COMPARISON
OF
GONADOSOMATIC
INDEX
(
GSI)
ACROSS
SAMPLING
AREAS
BY
SEX
295
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
F
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
NPAR1WAY
Procedure
Wilcoxon
Scores
(
Rank
Sums)
for
Variable
GSI
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
4
177.50
152.0
42.403752
44.375000
HL01
7
421.50
266.0
54.896998
60.214286
HL02
2
149.00
76.0
30.403357
74.500000
HW01
4
207.50
152.0
42.403752
51.875000
KZ03
13
469.50
494.0
71.435261
36.115385
KZ07
22
698.50
836.0
85.920025
31.750000
LPR0
23
726.50
874.0
87.018323
31.586957
Average
scores
were
used
for
ties.

Kruskal­
Wallis
Test
Chi­
Square
18.7491
DF
6
Pr
>
Chi­
Square
0.0046
Median
Scores
(
Number
of
Points
Above
Median)
for
Variable
GSI
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
4
3.0
1.973333
0.979433
0.750000
HL01
7
7.0
3.453333
1.267999
1.000000
HL02
2
2.0
0.986667
0.702250
1.000000
HW01
4
2.0
1.973333
0.979433
0.500000
KZ03
13
5.0
6.413333
1.649997
0.384615
KZ07
22
9.0
10.853333
1.984563
0.409091
LPR0
23
9.0
11.346667
2.009931
0.391304
Average
scores
were
used
for
ties.
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
18
of
41
Median
One­
Way
Analysis
Chi­
Square
12.3288
DF
6
Pr
>
Chi­
Square
0.0550
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
19
of
41
NONPARAMETRIC
COMPARISON
OF
GONADOSOMATIC
INDEX
(
GSI)
ACROSS
SAMPLING
AREAS
BY
SEX
297
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
M
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
NPAR1WAY
Procedure
Wilcoxon
Scores
(
Rank
Sums)
for
Variable
GSI
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
24
1407.50
1248.0
127.893490
58.645833
HL01
18
580.00
936.0
114.888084
32.222222
HW01
1
13.50
52.0
29.663976
13.500000
KZ03
9
541.50
468.0
85.430796
60.166667
KZ07
27
1675.50
1404.0
133.050941
62.055556
LPR0
24
1138.00
1248.0
127.893490
47.416667
Average
scores
were
used
for
ties.

Kruskal­
Wallis
Test
Chi­
Square
15.1001
DF
5
Pr
>
Chi­
Square
0.0099
Median
Scores
(
Number
of
Points
Above
Median)
for
Variable
GSI
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
24
14.0
11.883495
2.155602
0.583333
HL01
18
4.0
8.912621
1.936400
0.222222
HW01
1
0.0
0.495146
0.499976
0.000000
KZ03
9
6.0
4.456311
1.439908
0.666667
KZ07
27
17.0
13.368932
2.242529
0.629630
LPR0
24
10.0
11.883495
2.155602
0.416667
Average
scores
were
used
for
ties.

Median
One­
Way
Analysis
Chi­
Square
10.5912
DF
5
Pr
>
Chi­
Square
0.0601
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
20
of
41
AVERAGE
TESTOSTERONE
CONCENTRATIONS
BY
AREA
AND
SEX
COLLECTED
299
Obs
AREA
SEX
_
TYPE_
_
FREQ_
MEAN
STD
CV
MIN
MAX
1
HG01
F
0
9
149.68
98.03
65.496
49.87
307.18
2
HG01
M
0
38
513.61
1733.07
337.427
38.27
10715.78
3
HL02
F
0
2
2840.59
1307.26
46.021
1916.22
3764.96
4
HLO1
F
0
13
590.12
646.18
109.498
57.32
1970.54
5
HLO1
M
0
30
338.30
610.43
180.440
29.82
3068.27
6
HW01
F
0
4
648.93
533.77
82.254
73.90
1241.24
7
HW01
M
0
1
89.13
.
.
89.13
89.13
8
KZ02
F
0
16
4448.80
2370.51
53.284
1799.48
8897.31
9
KZ02
M
0
14
3504.74
1232.96
35.180
1281.92
5833.79
10
KZ03
F
0
17
1213.58
2101.09
173.132
14.26
8278.41
11
KZ03
M
0
13
604.88
887.89
146.788
24.32
2777.84
12
KZ07
F
0
22
226.46
115.27
50.901
58.45
407.51
13
KZ07
M
0
28
120.06
78.48
65.366
42.10
267.42
14
KZ07
U
0
1
54.52
.
.
54.52
54.52
15
LPR0
F
0
26
3377.91
3499.32
103.594
82.50
10347.86
16
LPR0
M
0
36
4061.89
4180.15
102.911
8.35
13154.07
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
21
of
41
ANALYSIS
OF
VARIANCE
FOR
TESTOSTERONE
CONCENTRATION
BETWEEN
SAMPLING
AREAS
BY
SEX
300
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
F
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
GLM
Procedure
Class
Level
Information
Class
Levels
Values
AREA
8
HG01
HL02
HLO1
HW01
KZ02
KZ03
KZ07
LPR0
Number
of
observations
109
NOTE:
Due
to
missing
values,
only
105
observations
can
be
used
in
this
analysis.

Dependent
Variable:
T
Sum
of
Source
DF
Squares
Mean
Square
F
Value
Pr
>
F
Model
7
277040769.7
39577252.8
8.19
<.
0001
Error
97
468931321.7
4834343.5
Corrected
Total
104
745972091.4
R­
Square
Coeff
Var
Root
MSE
T
Mean
0.371382
114.8531
2198.714
1914.371
Source
DF
Type
I
SS
Mean
Square
F
Value
Pr
>
F
AREA
7
277040769.7
39577252.8
8.19
<.
0001
Source
DF
Type
III
SS
Mean
Square
F
Value
Pr
>
F
AREA
7
277040769.7
39577252.8
8.19
<.
0001
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
22
of
41
ANALYSIS
OF
VARIANCE
FOR
TESTOSTERONE
CONCENTRATION
BETWEEN
SAMPLING
AREAS
BY
SEX
302
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
M
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
GLM
Procedure
Class
Level
Information
Class
Levels
Values
AREA
7
HG01
HLO1
HW01
KZ02
KZ03
KZ07
LPR0
Number
of
observations
160
NOTE:
Due
to
missing
values,
only
150
observations
can
be
used
in
this
analysis.

Dependent
Variable:
T
Sum
of
Source
DF
Squares
Mean
Square
F
Value
Pr
>
F
Model
6
416991901
69498650
13.08
<.
0001
Error
143
759843162
5313589
Corrected
Total
149
1176835063
R­
Square
Coeff
Var
Root
MSE
T
Mean
0.354333
147.3356
2305.122
1564.538
Source
DF
Type
I
SS
Mean
Square
F
Value
Pr
>
F
AREA
6
416991901.3
69498650.2
13.08
<.
0001
Source
DF
Type
III
SS
Mean
Square
F
Value
Pr
>
F
AREA
6
416991901.3
69498650.2
13.08
<.
0001
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
23
of
41
PROC
UNIVARIATE
OUTPUT
FOR
RESIDUALS
FROM
GLM
PROCEDURE
USING
TESTOSTERONE
DATA
BY
SEX
306
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
F
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
UNIVARIATE
Procedure
Variable:
Resid
Moments
N
105
Sum
Weights
105
Mean
0
Sum
Observations
0
Std
Deviation
2123.43001
Variance
4508955.02
Skewness
1.55804522
Kurtosis
3.17959841
Uncorrected
SS
468931322
Corrected
SS
468931322
Coeff
Variation
.
Std
Error
Mean
207.22555
Basic
Statistical
Measures
Location
Variability
Mean
0.000
Std
Deviation
2123
Median
­
100.921
Variance
4508955
Mode
.
Range
10360
Interquartile
Range
1365
Tests
for
Location:
Mu0=
0
Test
­
Statistic­
­­­­­
p
Value­­­­­­

Student's
t
t
0
Pr
>
|
t|
1.0000
Sign
M
­
12.5
Pr
>=
|
M|
0.0187
Signed
Rank
S
­
629
Pr
>=
|
S|
0.0438
Tests
for
Normality
Test
­­
Statistic­­­
­­­­­
p
Value­­­­­­

Shapiro­
Wilk
W
0.833207
Pr
<
W
<
0.0001
Kolmogorov­
Smirnov
D
0.220141
Pr
>
D
<
0.0100
Cramer­
von
Mises
W­
Sq
1.093384
Pr
>
W­
Sq
<
0.0050
Anderson­
Darling
A­
Sq
5.919474
Pr
>
A­
Sq
<
0.0050
Quantiles
(
Definition
5)

Quantile
Estimate
100%
Max
7064.836
99%
6969.954
95%
4998.822
90%
2711.678
75%
Q3
227.102
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
24
of
41
PROC
UNIVARIATE
OUTPUT
FOR
RESIDUALS
FROM
GLM
PROCEDURE
USING
TESTOSTERONE
DATA
BY
SEX
307
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
F
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
UNIVARIATE
Procedure
Variable:
Resid
Quantiles
(
Definition
5)

Quantile
Estimate
50%
Median
­
100.921
25%
Q1
­
1138.011
10%
­
2306.981
5%
­
3036.410
1%
­
3281.251
0%
Min
­
3295.404
Extreme
Observations
­­­­­­
Lowest­­­­­
­­­­­
Highest­­­­­

Value
Obs
Value
Obs
­
3295.40
84
5896.93
105
­
3281.25
89
6351.64
108
­
3251.36
91
6823.05
102
­
3196.13
86
6969.95
103
­
3142.45
87
7064.84
45
Missing
Values
­­­­­
Percent
Of­­­­­
Missing
Missing
Value
Count
All
Obs
Obs
.
4
3.67
100.00
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
F
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
UNIVARIATE
Procedure
Variable:
Resid
Stem
Leaf
#
Boxplot
7
01
2
*
6
8
1
*
6
4
1
*
5
9
1
*
5
0
1
*
4
4
14
2
0
3
9
1
0
3
2
78
2
0
2
0
1
|
1
9
1
|
1
44
2
|
0
556889
6
|
0
0000111112222233444
19
+­­+­­+
­
0
444433221111111111100000
24
*­­­­­*
­
0
997655555
9
|
|
­
1
332222222111000
15
+­­­­­+
­
1
776
3
|
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
25
of
41
­
2
3220
4
|
­
2
9655
4
|
­
3
333210
6
0
­­­­+­­­­+­­­­+­­­­+­­­­
Multiply
Stem.
Leaf
by
10**+
3
PROC
UNIVARIATE
OUTPUT
FOR
RESIDUALS
FROM
GLM
PROCEDURE
USING
TESTOSTERONE
DATA
BY
SEX
309
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
F
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
UNIVARIATE
Procedure
Variable:
Resid
Normal
Probability
Plot
7250+
*
|
*
*
|
*
|
*
|
++
|
*
++
|
*
+++
3750+
*
++
|
++
|
+*+
|
++*
|
++
*
|
+++
*
|
++
**
250+
+++******
|
*******
|
****
|
*****+
|
**++
|
***+
|
**++
­
3250+*
*
*
***+
+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+
­
2
­
1
0
+
1
+
2
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
26
of
41
PROC
UNIVARIATE
OUTPUT
FOR
RESIDUALS
FROM
GLM
PROCEDURE
USING
TESTOSTERONE
DATA
BY
SEX
310
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
M
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
UNIVARIATE
Procedure
Variable:
Resid
Moments
N
150
Sum
Weights
150
Mean
0
Sum
Observations
0
Std
Deviation
2258.2335
Variance
5099618.54
Skewness
2.16873075
Kurtosis
7.13432301
Uncorrected
SS
759843162
Corrected
SS
759843162
Coeff
Variation
.
Std
Error
Mean
184.383993
Basic
Statistical
Measures
Location
Variability
Mean
0.000
Std
Deviation
2258
Median
­
244.809
Variance
5099619
Mode
.
Range
14256
Interquartile
Range
458.79593
Tests
for
Location:
Mu0=
0
Test
­
Statistic­
­­­­­
p
Value­­­­­­

Student's
t
t
0
Pr
>
|
t|
1.0000
Sign
M
­
39
Pr
>=
|
M|
<.
0001
Signed
Rank
S
­
2514.5
Pr
>=
|
S|
<.
0001
Tests
for
Normality
Test
­­
Statistic­­­
­­­­­
p
Value­­­­­­

Shapiro­
Wilk
W
0.70622
Pr
<
W
<
0.0001
Kolmogorov­
Smirnov
D
0.307319
Pr
>
D
<
0.0100
Cramer­
von
Mises
W­
Sq
3.515333
Pr
>
W­
Sq
<
0.0050
Anderson­
Darling
A­
Sq
16.93825
Pr
>
A­
Sq
<
0.0050
Quantiles
(
Definition
5)

Quantile
Estimate
100%
Max
10202.1644
99%
9092.1757
95%
5619.6492
90%
1567.9838
75%
Q3
­
11.1053
50%
Median
­
244.8092
25%
Q1
­
469.9013
10%
­
2330.2361
5%
­
3256.9046
1%
­
3995.9020
0%
Min
­
4053.5445
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
27
of
41
Extreme
Observations
­­­­­­
Lowest­­­­­
­­­­­­
Highest­­­­­

Value
Obs
Value
Obs
­
4053.54
130
7257.12
144
­
3995.90
128
7952.52
141
­
3958.06
131
9020.27
159
­
3829.78
129
9092.18
145
­
3776.94
127
10202.16
37
Missing
Values
­­­­­
Percent
Of­­­­­
Missing
Missing
Value
Count
All
Obs
Obs
.
10
6.25
100.00
PROC
UNIVARIATE
OUTPUT
FOR
RESIDUALS
FROM
GLM
PROCEDURE
USING
TESTOSTERONE
DATA
BY
SEX
312
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
M
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
UNIVARIATE
Procedure
Variable:
Resid
Histogram
#
Boxplot
10500+*
1
*
.*
2
*
.
7500+*
2
*
.*
1
*
.*
2
*
4500+
.*
2
*
.**
4
*
1500+**
4
0
.*********
18
0
.**********************************************
91
+­­­­­+
­
1500+***
5
0
.*****
9
*
.****
8
*
­
4500+*
1
*
­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+­
*
may
represent
up
to
2
counts
Normal
Probability
Plot
10500+
*
|
*
*
|
7500+
**
|
*
|
**
+++
4500+
+++++
|
+**+
|
++++***
1500+
++++
**
|
+++++
******
|
******************
­
1500+
***
++++
|
****+++
|
*
******++
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
28
of
41
­
4500+*
+++++
+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+
­
2
­
1
0
+
1
+
2
PROC
UNIVARIATE
OUTPUT
FOR
RESIDUALS
FROM
GLM
PROCEDURE
USING
TESTOSTERONE
DATA
BY
SEX
315
The
UNIVARIATE
Procedure
Variable:
Resid
Schematic
Plots
|
10000
+
*
|
|
*
|
8000
+
*
|
*
|
*
|
*
*
6000
+
*
*
|
*
|
*
|
*
4000
+
0
*
|
*
|
0
*
|
0
*
2000
+
|
*
|
|
*
|
|
0
|
|
0
0
+
*­­+­­*
*­­+­­*
*­­+­­*
|
|
|
+­­­­­+
|
+­­­­­+
0
|
|
0
­
2000
+
|
0
|
|
*
|
0
*
|
0
*
­
4000
+
*
­­­­­­­­­­­­+­­­­­­­­­­­+­­­­­­­­­­­+­­­­­­­­­­­
SEX
F
M
U
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
29
of
41
NONPARAMETRIC
COMPARISON
OF
PLASMA
TESTOSTERONE
(
T)
CONCENTRATION
ACROSS
COLLECTION
AREAS
B
316
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
F
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
NPAR1WAY
Procedure
Wilcoxon
Scores
(
Rank
Sums)
for
Variable
T
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
9
213.0
477.0
87.361319
23.666667
HL02
2
159.0
106.0
42.657551
79.500000
HLO1
13
529.0
689.0
102.784564
40.692308
HW01
4
178.0
212.0
59.738318
44.500000
KZ02
16
1391.0
848.0
112.154655
86.937500
KZ03
17
659.0
901.0
114.955064
38.764706
KZ07
18
625.0
954.0
117.613775
34.722222
LPR0
26
1811.0
1378.0
134.698429
69.653846
Kruskal­
Wallis
Test
Chi­
Square
50.1393
DF
7
Pr
>
Chi­
Square
<.
0001
Median
Scores
(
Number
of
Points
Above
Median)
for
Variable
T
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
9
0.0
4.457143
1.441088
0.000000
HL02
2
2.0
0.990476
0.703667
1.000000
HLO1
13
5.0
6.438095
1.695506
0.384615
HW01
4
2.0
1.980952
0.985427
0.500000
KZ02
16
16.0
7.923810
1.850072
1.000000
KZ03
17
7.0
8.419048
1.896267
0.411765
KZ07
18
0.0
8.914286
1.940124
0.000000
LPR0
26
20.0
12.876190
2.221948
0.769231
Median
One­
Way
Analysis
Chi­
Square
53.2436
DF
7
Pr
>
Chi­
Square
<.
0001
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
30
of
41
NONPARAMETRIC
COMPARISON
OF
PLASMA
TESTOSTERONE
(
T)
CONCENTRATION
ACROSS
COLLECTION
AREAS
B
318
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
M
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
NPAR1WAY
Procedure
Wilcoxon
Scores
(
Rank
Sums)
for
Variable
T
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
37
2283.0
2793.50
229.370697
61.702703
HLO1
30
1665.0
2265.00
212.837967
55.500000
HW01
1
34.0
75.50
43.300308
34.000000
KZ02
14
1791.0
1057.00
154.785874
127.928571
KZ03
13
762.0
981.50
149.702761
58.615385
KZ07
19
734.0
1434.50
176.974339
38.631579
LPR0
36
4056.0
2718.00
227.248762
112.666667
Kruskal­
Wallis
Test
Chi­
Square
73.3827
DF
6
Pr
>
Chi­
Square
<.
0001
Median
Scores
(
Number
of
Points
Above
Median)
for
Variable
T
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
37
11.0
18.50
2.648604
0.297297
HLO1
30
9.0
15.00
2.457696
0.300000
HW01
1
0.0
0.50
0.500000
0.000000
KZ02
14
14.0
7.00
1.787353
1.000000
KZ03
13
6.0
6.50
1.728657
0.461538
KZ07
19
3.0
9.50
2.043569
0.157895
LPR0
36
32.0
18.00
2.624101
0.888889
Median
One­
Way
Analysis
Chi­
Square
56.2530
DF
6
Pr
>
Chi­
Square
<.
0001
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
31
of
41
AVERAGE
ESTRADIOL
CONCENTRATIONS
BY
AREA
AND
SEX
COLLECTED
320
Obs
AREA
SEX
_
TYPE_
_
FREQ_
MEAN
STD
CV
MIN
MAX
1
HG01
F
0
9
136.09
171.55
126.061
18.88
541.24
2
HG01
M
0
38
1350.88
2956.38
218.848
2.50
15789.18
3
HL01
F
0
13
239.70
182.96
76.328
26.92
497.18
4
HL01
M
0
30
916.63
3825.62
417.356
25.33
18036.73
5
HL02
F
0
2
805.78
541.62
67.217
422.80
1188.77
6
HW01
F
0
4
129.45
225.55
174.237
1.11
466.49
7
HW01
M
0
1
25.79
.
.
25.79
25.79
8
KZ03
F
0
17
3222.80
4573.00
141.895
4.42
16236.64
9
KZ03
M
0
13
4821.49
7678.18
159.249
52.64
21183.02
10
KZ07
F
0
22
463.71
478.15
103.113
10.39
1887.19
11
KZ07
M
0
28
582.12
1199.15
205.997
6.27
5610.75
12
KZ07
U
0
1
174.92
.
.
174.92
174.92
13
KZO2
F
0
16
10770.41
4310.62
40.023
2744.22
19133.06
14
KZO2
M
0
14
10490.20
2634.88
25.118
6449.99
16600.01
15
LPR0
F
0
26
4599.44
4687.58
101.916
1.21
16969.95
16
LPR0
M
0
36
4739.09
3926.58
82.855
1.10
13756.41
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
32
of
41
ANALYSIS
OF
VARIANCE
FOR
ESTRADIOL
CONCENTRATION
BETWEEN
SAMPLING
AREAS
BY
SEX
321
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
F
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
GLM
Procedure
Class
Level
Information
Class
Levels
Values
AREA
8
HG01
HL01
HL02
HW01
KZ03
KZ07
KZO2
LPR0
Number
of
observations
109
NOTE:
Due
to
missing
values,
only
105
observations
can
be
used
in
this
analysis.
Dependent
Variable:
E2
Sum
of
Source
DF
Squares
Mean
Square
F
Value
Pr
>
F
Model
7
1338518159
191216880
15.87
<.
0001
Error
97
1168404398
12045406
Corrected
Total
104
2506922556
R­
Square
Coeff
Var
Root
MSE
E2
Mean
0.533929
100.5534
3470.649
3451.549
Source
DF
Type
I
SS
Mean
Square
F
Value
Pr
>
F
AREA
7
1338518159
191216880
15.87
<.
0001
Source
DF
Type
III
SS
Mean
Square
F
Value
Pr
>
F
AREA
7
1338518159
191216880
15.87
<.
0001
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
33
of
41
ANALYSIS
OF
VARIANCE
FOR
ESTRADIOL
CONCENTRATION
BETWEEN
SAMPLING
AREAS
BY
SEX
323
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
M
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
GLM
Procedure
Class
Level
Information
Class
Levels
Values
AREA
7
HG01
HL01
HW01
KZ03
KZ07
KZO2
LPR0
Number
of
observations
160
NOTE:
Due
to
missing
values,
only
148
observations
can
be
used
in
this
analysis.

Dependent
Variable:
E2
Sum
of
Source
DF
Squares
Mean
Square
F
Value
Pr
>
F
Model
6
1292294626
215382438
15.34
<.
0001
Error
141
1979233920
14037120
Corrected
Total
147
3271528546
R­
Square
Coeff
Var
Root
MSE
E2
Mean
0.395012
119.6745
3746.614
3130.671
Source
DF
Type
I
SS
Mean
Square
F
Value
Pr
>
F
AREA
6
1292294626
215382438
15.34
<.
0001
Source
DF
Type
III
SS
Mean
Square
F
Value
Pr
>
F
AREA
6
1292294626
215382438
15.34
<.
0001
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
34
of
41
PROC
UNIVARIATE
OUTPUT
FOR
RESIDUALS
FROM
GLM
PROCEDURE
USING
ESTRADIOL
DATA
BY
SEX
327
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
F
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
UNIVARIATE
Procedure
Variable:
Resid
Moments
N
105
Sum
Weights
105
Mean
0
Sum
Observations
0
Std
Deviation
3351.81409
Variance
11234657.7
Skewness
1.5010092
Kurtosis
4.28501184
Uncorrected
SS
1168404398
Corrected
SS
1168404398
Coeff
Variation
.
Std
Error
Mean
327.103561
Basic
Statistical
Measures
Location
Variability
Mean
0.000
Std
Deviation
3352
Median
­
123.888
Variance
11234658
Mode
.
Range
21040
Interquartile
Range
2367
Tests
for
Location:
Mu0=
0
Test
­
Statistic­
­­­­­
p
Value­­­­­­

Student's
t
t
0
Pr
>
|
t|
1.0000
Sign
M
­
12.5
Pr
>=
|
M|
0.0187
Signed
Rank
S
­
426
Pr
>=
|
S|
0.1744
Tests
for
Normality
Test
­­
Statistic­­­
­­­­­
p
Value­­­­­­

Shapiro­
Wilk
W
0.860111
Pr
<
W
<
0.0001
Kolmogorov­
Smirnov
D
0.213219
Pr
>
D
<
0.0100
Cramer­
von
Mises
W­
Sq
0.890385
Pr
>
W­
Sq
<
0.0050
Anderson­
Darling
A­
Sq
4.5607
Pr
>
A­
Sq
<
0.0050
Quantiles
(
Definition
5)

Quantile
Estimate
100%
Max
13013.841
99%
12370.510
95%
5876.148
90%
3773.767
75%
Q3
486.776
50%
Median
­
123.888
25%
Q1
­
1880.503
10%
­
3569.116
5%
­
4370.211
1%
­
4598.227
0%
Min
­
8026.193
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
35
of
41
Extreme
Observations
­­­­­­
Lowest­­­­­
­­­­­­
Highest­­­­­

Value
Obs
Value
Obs
­
8026.19
68
6199.51
105
­
4598.23
89
8362.65
74
­
4598.05
88
11923.26
102
­
4531.09
90
12370.51
103
­
4436.59
77
13013.84
40
Missing
Values
­­­­­
Percent
Of­­­­­
Missing
Missing
Value
Count
All
Obs
Obs
.
4
3.67
100.00
PROC
UNIVARIATE
OUTPUT
FOR
RESIDUALS
FROM
GLM
PROCEDURE
USING
ESTRADIOL
DATA
BY
SEX
329
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
F
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
UNIVARIATE
Procedure
Variable:
Resid
Stem
Leaf
#
Boxplot
13
0
1
*
12
4
1
*
11
9
1
*
10
9
8
4
1
*
7
6
2
1
0
5
29
2
0
4
356
3
0
3
23558
5
|
2
24
2
|
1
01467
5
|
0
111222233344455567
18
+­­+­­+
­
0
9765544444433322221111111111111110
34
*­­­­­*
­
1
97653
5
+­­­­­+
­
2
88877741
8
|
­
3
764422210
9
|
­
4
66544422
8
|
­
5
­
6
­
7
­
8
0
1
0
­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­
Multiply
Stem.
Leaf
by
10**+
3
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
36
of
41
PROC
UNIVARIATE
OUTPUT
FOR
RESIDUALS
FROM
GLM
PROCEDURE
USING
ESTRADIOL
DATA
BY
SEX
330
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
F
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
UNIVARIATE
Procedure
Variable:
Resid
Normal
Probability
Plot
13500+
*
|
*
|
*
|
|
|
*
++
|
+++
|
*+++
|
**+
|
+***
|
++***
2500+
+++
*
|
+++
**
|
+++******
|
*********
|
**+++
|
***+
|
****+
|
*
*
*****++
|
+++
|
+++
|
+++
­
8500+*+
+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+
­
2
­
1
0
+
1
+
2
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
37
of
41
PROC
UNIVARIATE
OUTPUT
FOR
RESIDUALS
FROM
GLM
PROCEDURE
USING
ESTRADIOL
DATA
BY
SEX
331
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
M
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
UNIVARIATE
Procedure
Variable:
Resid
Moments
N
148
Sum
Weights
148
Mean
0
Sum
Observations
0
Std
Deviation
3669.35639
Variance
13464176.3
Skewness
2.47973855
Kurtosis
8.42158109
Uncorrected
SS
1979233920
Corrected
SS
1979233920
Coeff
Variation
.
Std
Error
Mean
301.619237
Basic
Statistical
Measures
Location
Variability
Mean
0.000
Std
Deviation
3669
Median
­
762.279
Variance
13464176
Mode
.
Range
21889
Interquartile
Range
1179
Tests
for
Location:
Mu0=
0
Test
­
Statistic­
­­­­­
p
Value­­­­­­

Student's
t
t
0
Pr
>
|
t|
1.0000
Sign
M
­
38
Pr
>=
|
M|
<.
0001
Signed
Rank
S
­
1821
Pr
>=
|
S|
0.0004
Tests
for
Normality
Test
­­
Statistic­­­
­­­­­
p
Value­­­­­­

Shapiro­
Wilk
W
0.727461
Pr
<
W
<
0.0001
Kolmogorov­
Smirnov
D
0.256757
Pr
>
D
<
0.0100
Cramer­
von
Mises
W­
Sq
2.560058
Pr
>
W­
Sq
<
0.0050
Anderson­
Darling
A­
Sq
12.75496
Pr
>
A­
Sq
<
0.0050
Quantiles
(
Definition
5)

Quantile
Estimate
100%
Max
17120.0961
99%
16361.5293
95%
6152.1951
90%
3682.6693
75%
Q3
­
17.4236
50%
Median
­
762.2792
25%
Q1
­
1196.6524
10%
­
4040.2126
5%
­
4729.7052
1%
­
4737.9875
0%
Min
­
4768.8486
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
38
of
41
Extreme
Observations
­­­­­­
Lowest­­­­­
­­­­­­
Highest­­­­­

Value
Obs
Value
Obs
­
4768.85
70
9017.32
156
­
4737.99
126
14438.29
37
­
4737.71
128
15862.95
80
­
4736.44
130
16361.53
79
­
4736.35
129
17120.10
47
Missing
Values
­­­­­
Percent
Of­­­­­
Missing
Missing
Value
Count
All
Obs
Obs
.
12
7.50
100.00
PROC
UNIVARIATE
OUTPUT
FOR
RESIDUALS
FROM
GLM
PROCEDURE
USING
ESTRADIOL
DATA
BY
SEX
333
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
M
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
UNIVARIATE
Procedure
Variable:
Resid
Histogram
#
Boxplot
17500+*
1
*
.*
1
*
.*
1
*
.*
1
*
.
.
.
.
.*
1
*
.*
1
*
.*
1
*
6500+**
3
*
.*
2
*
.*
2
*
.**
4
0
.****
8
0
.*
2
0
.*****
9
+
.***************************
54
+­­­­­+
.******************
35
+­­­­­+
.***
5
|
.*
1
0
­
4500+********
16
0
­­­­+­­­­+­­­­+­­­­+­­­­+­­
*
may
represent
up
to
2
counts
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
39
of
41
PROC
UNIVARIATE
OUTPUT
FOR
RESIDUALS
FROM
GLM
PROCEDURE
USING
ESTRADIOL
DATA
BY
SEX
334
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
M
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
UNIVARIATE
Procedure
Variable:
Resid
Normal
Probability
Plot
17500+
*
|
*
|
*
|
*
|
|
|
|
|
*
+
|
*
+++
|
*
++
6500+
**+++
|
**+
|
++**
|
++
**
|
+++***
|
+++
*
|
+++
***
|
***********
|
********
|
**
+++
|
*
++
­
4500+*
*
*********++
+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+­­­­+
­
2
­
1
0
+
1
+
2
|
0
*
5000
+
0
*
|
0
|
|
*
|
|
0
2500
+
|
0
|
|
0
|
|
|
|
+­­­­­+
|
0
+
*­­+­­*
+­­+­­+
*­­+­­*
|
|
|
*­­­­­*
|
|
|
+­­­­­+
|
+­­­­­+
|
­
2500
+
|
|
|
|
0
|
|
0
|
|
0
­
5000
+
0
|
|
|
­
7500
+
|
0
|
|
­
10000
+
­­­­­­­­­­­­+­­­­­­­­­­­+­­­­­­­­­­­+­­­­­­­­­­­
SEX
F
M
U
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
40
of
41
NONPARAMETRIC
COMPARISON
OF
PLASMA
ESTRADIOL
CONCENTRATION
ACROSS
COLLECTION
AREAS
BY
SEX
338
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
F
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
NPAR1WAY
Procedure
Wilcoxon
Scores
(
Rank
Sums)
for
Variable
E2
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
9
202.0
477.0
87.361319
22.444444
HL01
9
296.0
477.0
87.361319
32.888889
HL02
2
109.0
106.0
42.657551
54.500000
HW01
4
71.0
212.0
59.738318
17.750000
KZ03
17
910.0
901.0
114.955064
53.529412
KZ07
22
838.0
1166.0
127.002625
38.090909
KZO2
16
1467.0
848.0
112.154655
91.687500
LPR0
26
1672.0
1378.0
134.698429
64.307692
Kruskal­
Wallis
Test
Chi­
Square
53.0292
DF
7
Pr
>
Chi­
Square
<.
0001
Median
Scores
(
Number
of
Points
Above
Median)
for
Variable
E2
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
9
0.0
4.457143
1.441088
0.000000
HL01
9
0.0
4.457143
1.441088
0.000000
HL02
2
1.0
0.990476
0.703667
0.500000
HW01
4
0.0
1.980952
0.985427
0.000000
KZ03
17
8.0
8.419048
1.896267
0.470588
KZ07
22
8.0
10.895238
2.095000
0.363636
KZO2
16
16.0
7.923810
1.850072
1.000000
LPR0
26
19.0
12.876190
2.221948
0.730769
Median
One­
Way
Analysis
Chi­
Square
44.7975
DF
7
Pr
>
Chi­
Square
<.
0001
Data
Evaluation
Report
on
Tissue
Pesticide
Residues
and
Histology
of
the
Larynx
and
Gonads
in
Native
Green
Frogs
(
Rana
clamitans)
Collected
from
Agricultural
Areas
in
Michigan:
Hormone
Analysis.
EPA
MRID
Number
458677­
02
Page
41
of
41
NONPARAMETRIC
COMPARISON
OF
PLASMA
ESTRADIOL
CONCENTRATION
ACROSS
COLLECTION
AREAS
BY
SEX
340
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­
SEX=
M
­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­

The
NPAR1WAY
Procedure
Wilcoxon
Scores
(
Rank
Sums)
for
Variable
E2
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
35
2434.0
2607.50
221.603061
69.542857
HL01
22
1021.0
1639.00
185.523583
46.409091
HW01
1
23.0
74.50
42.722945
23.000000
KZ03
13
1070.0
968.50
147.618596
82.307692
KZ07
27
1160.0
2011.50
201.408168
42.962963
KZO2
14
1856.0
1043.00
152.622628
132.571429
LPR0
36
3462.0
2682.00
223.749860
96.166667
Kruskal­
Wallis
Test
Chi­
Square
61.2901
DF
6
Pr
>
Chi­
Square
<.
0001
Median
Scores
(
Number
of
Points
Above
Median)
for
Variable
E2
Classified
by
Variable
AREA
Sum
of
Expected
Std
Dev
Mean
AREA
N
Scores
Under
H0
Under
H0
Score
ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ
HG01
35
11.0
17.50
2.593490
0.314286
HL01
22
3.0
11.00
2.171241
0.136364
HW01
1
0.0
0.50
0.500000
0.000000
KZ03
13
6.0
6.50
1.727627
0.461538
KZ07
27
9.0
13.50
2.357143
0.333333
KZO2
14
14.0
7.00
1.786190
1.000000
LPR0
36
31.0
18.00
2.618615
0.861111
Median
One­
Way
Analysis
Chi­
Square
52.9594
DF
6
Pr
>
Chi­
Square
<.
0001