Professional Documents
Culture Documents
Biologia
Biologia
of Natural
1School
BACKGROUND: Many
evidence of endocrine
and
sial. Typically,
studies
sites.
unpolluted
and department
Heather
of Zoology,
of Florida,
University
gonadal
EDC-induced
to address
the rela
this question
by explicidy
quantifying
and habitats characterized
by differing
degrees of agricul
ductive
areas
to
exposed
secondary
11536
sexual
to
which
(EDCs),
endocrine-disrupting
alter normal
hormonal
morphology
Zal?
chemicals
signal
can per
so EDCs
conserved,
highly
can
across
functions
physiological
affect
that
taxa
2006; McLachlan
2001).
tem
in a wide
function
array
of vertebrate
(Hayes
amphibians
2003),
reptiles
(Sonne
al.
2006),
documenting
natural
process.
This
controversy
is fueled,
in
effects
field
identified
observations.
Wild
in
and
generate
controlled
novel
patterns
of
toxic
particular
not
pathologies
studies.
laboratory
pathologies
because many
agent
identified
or
in
Investigating
in nature
is further
dreds of miles
Superior)
Thurman
humans
including
can
abnormalities
normal
diverse
to
of
complex mixtures
exposed
et al. 2006)
and other envi
(Hayes
et al. 2006)
conditions
(Edwards
a causal
link between
obfuscate
ronmental
2004). Moreover,
many
are
the endocrine
system
of
are
organisms
chemicals
and Penn
components
natural
gonadal
malformations
contaminants
(including
processes.
found no
atrazine)
and
argue
result
For
con
agricultural
and the inci
or testicular
hermaphroditism
In
with
testicular
their
oocytes.
study,
frogs
at both
found
oocytes were
(pol
agricultural
sites
and nonagricultural
luted)
(reference)
of
dence
are
et al. (2006)
by Murphy
reported
or to
to be
naturally
hermaphroditic
as adults
have
testicular
oocytes
commonly
2001;
observed
such
species
known
embryonic
development,
adult
system
reproductive
change
as well as
and function,
reproduc
ing during
manently
of
and
via
Exposure
that
from
found
agriculture
and they
developmental
et al. (2006)
instance, Murphy
sistent
between
relationship
traits.
available
not
have
between
relationships
abnormalities,
reproductive
intersex,
pesticides,
disruption,
116:1526-1532
(2008).
doi:10.1289/ehp.
amphibians,
Health
Perspect
Environ
decline
global
Indeed, Davidson
researchers
strong
sexual
endocrine
that
abnormalities
reproductive
to the
populations.
Other
contaminants.
agricultural
KEY WORDS:
from
relative
ductive
USA
func
and measures
of gonadal
of gonadal abnormalities
the occurrence
quantified
tion in at least 20 giant toads (Bufo marinus) from each of five sites that occur along a gradient of
land use from 0 to 97%.
increasing agricultural
concentrations
and secondary
hormone
activity and success, so affected toads likely have reduced
to amphibian
contribute
abnormalities
could certainly
contribute
amphibian
METHODS: We
Steroid
Florida,
Gainesville,
could
approach
abnormalities
Jr.,1'2
and from
those
collected
laboratory
areas
atrazine-contaminated
agricultural
to reference
and
sites,
they suggested
However,
systems of wildlife.
areas has been controver
in
living
agricultural
on wildlife
to compare polluted with
attempt
tural activity.
CONCLUSIONS:
Louis J. Guillette
J. Hamlin,2
endocrine
disrupt
in wild amphibians
disruption
on the effects of
pollutants
took a novel
OBJECTIVES: We
tionship between
M. Campbell,2
Chelsey
Environment,
contaminants
agricultural
Gonadal
Alters
Agriculture
(Nations
and Hallberg
1992;
and Cromwell
2000).
and
laboratory
recent
Many
have
that agricultural
suggested
are associated
with
amphibian
and
abnormalities
population
field
studies
contaminants
reproductive
declines
of gonadal
abnormali
incidences
higher
male
and
ties and
(intersex:
hermaphroditism
in
tissue in the same individual)
female gonadal
to the herbicide
in the
atrazine
frogs exposed
found
1526
volume
that 25-50%
of adults from three dif
reported
ferent species had testicular oocytes.
Such pro
in naturally
of testicular
oocytes
portions
gonochoristic
result of a rare
the
sites
Murphy
adults
are not
genetic
anomaly.
as
nonagricultural
were
nevertheless
categorized
et al. (2006)
to be the
likely
It is likely that
by
pol
to K.A. McCoy,
Smithsonian
correspondence
Research
Institute, Unit 0948, APO AA
Tropical
E-mail:
34002 USA. Telephone:
507-6-461-0821.
kristam@bu.edu
L. Babonis, and R. Harris
We
thank M. McCoy,
and
for constructive
feedback on the manuscript
A. Amick, P. Barnes Jr., M. Fromowitz, M. McCoy,
M. Rehrer, V. Bender, K. Hopkins,
and H. Wineman
Address
2008
accepted
Environmental
3 July 2008.
Health
Perspectives
Gonadal
we
took a novel
present
study,
to
gross
asking whether
gonadal
of an anuran amphibian
species
are associated with
{Bufo marinus)
agricultural
most
Unlike
studies, we
exposure.
previous
across a range of sites
this
question
explored
that vary in degree
of agricultural
intensity.
In
the
approach
abnormalities
at our
the proportion
of agriculture
was
to the
sites
related
inversely
study
degree
Stressors.
of urbanization
and its associated
In fact,
we
explicitly
are
abnormalities
Therefore,
gonadal
uted across
are
tested
with
distrib
randomly
human-modified
associated
whether
to
across
sites
five
We
found
and
types of
incidence
as well
as the
across
this continuum.
in
tural
to the
gonads
relative
of
nonagricul
sex
compared
circulating
and as more
concentrations,
integrated
toads. We
steroid
endocrine
system function,
signals of seasonal
we
three sex-hormone-dependent
compare
sexual characteristics.
secondary
Materials
and Methods
of
intensity
we
exposure,
agricultural
and
into
the
program
analysis
Pro
Image
Plus
House
(Image
image
A/S,
(Crosbie
Twenty
body
each
effect
in more
streetlights.
an overdose
with
B. marinus
et al.
is approximately
2 km2
(Zug
so this measure
reflects the area experi
1975),
enced by collected
toads.
sites an
assigned
"agricultural
on the
rank
of
based
1-5
intensity"
percentage
area around
in the 5.6-km2
of agriculture
the
then
site or
its proximity
to
agriculture
when
no
used
these
tau
of association.
Lake Worth
(LW) and
no
contained
(WT)
Wellington
agriculture
(0%). LW was 22 km away from an agricul
tural area and was ranked
was
1, whereas WT
5.18
ranked
km
and was
agriculture,
34%
(HS) contained
from
away
2. Homestead
agricultural
land, which
we
ranked
3. Canal
Environmental
Health
Perspectives
volume
of
at
at
collected
tri
0.3%
7
pH
(Sigma
and
photographed,
Aldrich,
measured
from
mercury
vapor
was
euthanized
the anesthetic
methanesulfonate
length
(nearest
via
collected
trifuged.
cardiac
Plasma
samples
back to the
transported
and stored
at -80?C
until
puncture
were frozen
laboratory
hormone
and
cen
at -20?C,
on
dry ice,
assays were
One
We
were
animals
D699);
treated
(ANOVA)
humanely,
and
characteristics.
all primary
sexual characteristics
individuals
First, we evaluated
in two ways.
each abnormality
Kruskal-Wallis
We
under
feeding,
Each
individual
type
across
sites with
of variance
analysis
if the occurrence
of
one-way
to determine
individuals
based on
categorized
We
type of gonadal
abnormality.
scores and evaluated
them based on
also
quantity
and
assigned
the total
number
of abnormalities
a measure
is considered
intersexed
tau tests
in R
free
statistics
IL),
soft
in females,
gonadal
morphology
males
with
Bidder's
gross
morphologic
so all
on
analyses
include
males,
only
abnormalities
organ
and intersexes.
These
toads
(Bidder's males),
=
were
a sex
1,
(males
group category
assigned
=
=
Bidder's males
and
differ
2, intersexes
3),
ences
in the mean
evaluated
as described
the
to increased
corresponded
agricul
an
As
of our ranking
intensity.
example
an individual
testes
with misshaped
system,
a
in the testes
(one abnormality
category),
Bidder's
(one
organ
abnormality
vitellogenic
and
sites
in the Bidder's
the presence
(two
genic
organ
of
received
category)
and ovaries
(in
category),
that were
also vitello
testes)
abnormalities
in the ovary/oviduct
a score of 4.
next
Our
of gonadal
Analyses
function.
was
to determine
if
from
the
toads
objective
areas had altered
most
agricultural
gonadal
relative to those from
function
nonagricultural
areas
to assess whether
intersex
and
and
Bidder's
males
from
males
the
from
nonagricultural
groups occurred
at
occurred
not
agricultural
demasculinized
and
feminized
compare
same
areas.
site
and
sites were
to
relative
those
from
at every
sites), so we could
across all sites.
groups
nonagricultural
gonadal
for each measure
of
func
gonadal
sex
toads into six different
grouped
on
based
gross gonadal morphology
categories
in
and their occurrence
(LW,
nonagricultural
Therefore,
tion, we
females
and/)
did
not
from
toads
the
intermediate
site
in these
(HS, 34%)
could not be considered
or as
AgMs.
NAgM
Sex hormone
concentrations.
We
analyzed
testosterone
and
(E2)
a validated
radio
17?-estradiol
circulating
concentrations
with
immunoassay
using
the 96-well
FlashPlate
observed
abnormalities
of
organs
tural
and Kendall's
severity
c) Bidder's
as
to
test whether
site
explicitly
corre
in sex group
statistically
the
In addition,
the total number
of abnor
were
summed
evaluated
and similarly
to determine
num
if the mean
and maximum
test of association
We
of
and
condition),
differences
1988).
of
three
tissue
a) testes,
types:
following
in the presence
of testes
b) ovaries/oviducts
and developmental
(number
stage of female
in the presence
tissues found
of testes
gonadal
agricultural
intensity
analyses because
they
Castellan
in each
the
malities
1993;
collected
than 1 year.
all locations were
from
while
night,
caine
were
We
agriculture
(Figure 1).
Year had no
period.
or
types of abnormal
at sites where
collections
ities documented
occurred
and
total percentage
of agricul
tural land
and sugarcane
(vegetable
farming,
no livestock)
a 5.6-km2
area around
within
each
collection
site. The
home
of
range
et al. 2006)
site over a 2-year
on the
percentage
with
the
of
with
pesti
typically
polluted
and Chow-Fraser
1999).
or more
adult
toads
(> 91 mm
length)
suites
(Standley
areas are more
conducted.
sur
sites and collection
We
Study
techniques.
that varied
in
veyed five sites in south Florida
to
habitat
suburban
type from
completely
To quantify
the
completely
agricultural.
different
induce
con
Polyaromatic
hydrocarbon
are
areas
in urbanized
ubiquitous
et al. 2000),
whereas
agricultural
pathologies.
centrations
Toads
abnor
gonadal
as
so
malities
increased,
activity
agricultural
we went
to determine
if the
further
of
gonads
areas had altered
in
toads
agricultural
living
function
increase
strong
4 and
to
expected
Meshaka
specifically.
agriculture
We
B. marinus
surveyed
abnormalities
gonadal
of intersex
individuals
are
cides
exposure
increasing
land;
agricultural
5, respectively.
the presence
of specific contami
Although
nants at each type of site
certainly
overlapped,
are
and agricultural
urbanized
habitats
pol
of chemicals
luted with
different
milieus
that
or
landscapes
the most
contained
97%)
these were
ranked
abnormalities
sex group
above.
2008
1527
across
sites were
were
to meet
the
transformed
log10
of normality
and
of
assumptions
homogeneity
a linear mixed
variances and then analyzed
using
where
and assay by sex
model,
assay
(plate)
Data
interaction
We
used
were
SPSS
identified
default
as the random
post
hoc
effects.
comparisons
(least significant
difference).
E2 to testosterone
were
ratios (E2/testosterone)
log10 transformed
to meet
the assumptions
of normality
and
^^H
H?
McCoy et al.
of variances
homogeneity
We
ANOVA.
one-way
comparisons
Secondary
and
evaluated
conducted
test.
using Tukey's
sexual characteristics.
to compare
photographs
number
of nuptial
pads,
using
hoc
post
relation
We
were
We
used
identified
renamed
blind
toads were
collected.
and
above.
observers
the
all statistics
using Tukey's
tests.
the number
pattern. We
quantified
across
that occurred
changes
(mottling)
a transect
line drawn
from the left eye to the
vent of each
toad using
Image
photographed
Color
of color
sex
across
Plus and compared
mottling
In addition,
groups using a one-way ANOVA.
we
scores
and female mottling
analyzed male
across sites
a two-way ANOVA
separately using
to test for
across
in color
dimorphism
changes
sites (sex X site).
Pro
size. We
Forelimb
across
the forelimb
hum?rus
using
the width
perpendicular
Pro Plus and tested
an
using
arm
to the
of
to the
distal
radioulna
and
Image
sex groups
among
measured
the
axis
for differences
of covari
analysis
were
slopes
homogeneous;
the interaction
from
the
thus, we
removed
means
marginal
group. We
of each
snout-vent
of
length
the elbow
compared
120.2 mm.
pads. No
Nuptial
for
sex groups
width
at a
Results
sexual
Primary
rank increased
characteristics.
as a function
sex group
Mean
of
agriculture.
two most
agricultural
sites
other
(Kruskal-Wallis4
=
=
1, p
0.01;
2A).
tau-b3
Figure
Testicular
abnormalities
IB), such as
(Figure
common
and
testes, were
relatively
misshaped
did not vary across sites or increase with
agri
=
=
cultural
5.1, p
0.272;
(KW4
intensity
=
=
Kendall's
the
0.4, p
0.23). However,
tau-b3
Kendall's
number
abnormalities
of female-tissue
or oviducts
ovaries
in the presence
different
among
were
(e.g.,
of
testes)
sites and
significantly
increased with
intensity
increasing
agricultural
=
=
=
Kendall's
17.3, p
0.002;
(KW4
tau-b3
=
0.9, p
0.02;
IC, 2B).
Figures
Importantly,
Bidder's
abnormalties
the number
and developmental
stage of female
in the presence
found
of testes
gonadal
are considered
a measure
of the severity of the
tissues
intersexed
of gonadal phenotypes
observed. Abbreviations:
B, Bidder's organ; C, con
Figure 1. Four broad categories
nective tissue; K, kidney; OV, ovary; OD, oviduct; T, testis. (A) Toads that had no obvious gross gonadal
as having normal gonadal morphology.
abnormalities were categorized
(Left) "Males" had testes, a non
Bidder's organ, and no ovaries or oviducts.
vitellogenic
(Right) "Females" had ovaries, oviducts, nonvitel
logenic Bidder's organs, and no testes; ovaries in various stages of oog?nesis were collected
(oog?nesis
as "male" in
and accumulation
of vitellogenin
increases from left to right). {B) Toads that were categorized
some cases had testicular abnormalities.
(Left) Multiple testes.
(Center) Abnormally shaped testes.
(Right)
Abnormally small testes. (0 Intersex toads had both testes and oviducts or ovaries. (Left)Well-developed
oviduct and testis, but no ovary. (Center) Nonvitellogenic
ovary that is distinct from the Bidder's organ and
testis. (Right) Intersex with a highly vitellogenic
ovary. {D) "Bidder's males" had multiple (left), early vitel
= 0.5 cm.
logenic (center), or late vitellogenic
(right) Bidder's organs but no ovaries or oviducts. Bars
1528
volume
toads
condition.
became
more
culture
increased.
tended
to vary
the
agricultural
that measure,
Applying
as
intersexed
severely
agri
Bidder's
organ abnormalities
across
sites
sites
(KW4
and was
=
9.0,
highest
=
0.06;
at
with
p<
(KW4
increasing
agriculture
=
Kendall's
0.001; mean
0.8,
tau-b3
2008
Environmental
Health
Perspectives
Gonadal
maximum
0.04;
Kendall's
tau-b3
0.9,
We
found
E2 concentra
=
<
sex
23.74,
groups
p
(^5,157
were
these differences
driven
in plasma
differences
significant
tions among
but
in females
by high E2 concentrations
sex
to all other
3A).
groups
(Figure
were not
differ
and NAgFs
AgFs
significantly
ent from one another.
entirely
relative
testosterone
Plasma
also
significantly
were
concentrations
different
among
sex
groups
=
<
(^5,162 1 !-26>P ?-001; Figure 3B). NAgFs
testos
in
did not differ
and AgFs
circulating
terone concentrations,
but had
significantly
testosterone
levels than did NAgMs.
lower
concen
testosterone
Intersex toads had plasma
trations
similar to NAgFs
and were
that were
not
from
different
Bidder's
but were
males
and AgMs,
lower
significantly
males
Bidder's
Both
0.001),
and AgMs
not
than NAgMs.
were
signifi
significantly
tive toNAgMs
The
ratio
testosterone
lower
levels
rela
concen
to testosterone
was
(E2/testosterone)
significantly
=
sex
different
48.46,
groups
among
(^5,150
< 0.001;
were
not
Females
3C).
p
sig
Figure
tration
nificantly
from
different
E2/testosterone
The
groups.
intersex
toads was
one
another
ratios
E2/testosterone
that
significantly
of females
Bidder's
groups
males
and
AgMs,
than NAgMs.
On
average,
more
E2 than testosterone
but had
to all
relative
higher
other
in both
different
significantly
from
Color
pattern.
of sex
function
ratio
lower
and
but was
intersex
of
than
similar
to
higher
toads had
(mean E2/testos
terone
males
ratio > 1; Figure
3C). Bidder's
were
between
and
and AgMs
intermediate
mottled
highly
Bidder's
males
females
and
NAgMs.
females
intersexes
score
Mottling
category
(/^
and
similar.
similar
<
in
mottling
mottled
more
significantly
were
as a
varied
=
15.6,
128
Intersexes
and
were
AgMs
but more
than
than
less mottled
mottled
than
NAgMs.
were
the only group
that was signifi
than all other sex groups.
less mottled
NAgMs
cantly
We
found
sex and
between
SHI
agriculture
(Figure 3C).
NAgMs
(Figure 3B).
of E2
abnormalities
a
significant
site on mottling
interaction
=
2.7,
Cr\io3
- 0.03;
Figure 4C). Although female mot
across
sites,
change
in
became
mottling
not
did
tling
the
sexual
less pro
increased
dimorphism
as
nounced
intensity
agricultural
were
in coloration
males
feminized
because
(Figure4C).
Forelimb size. Mean forelimb width was
different
significantly
sex groups
among
(F^jo
= 33.8,
p < 0.001; Figure 4D). Females had
smaller
forelimbs
Intersex
individuals
any other
group.
arm widths
that were
between
intermediate
ent from
than
had
females
and
significantly
and NAgMs.
Bidder's
females
not
significantly
from
NAgMs.
different
significantly
and
or
intersexes
pads. Nuptial
Nuptial
males
than did
between
intermediate
different
differ
among
sex groups
(we
6.0,
0.001;
4E).
Figure
Intersex
toads
numbers
of nuptial
pads
were
inter
and AgMs
and not
between
differ
significantly
ent from those of either intersex or
NAgMs.
NAgMs,
Bidder's
among
mediate
LW WT
HS
Increasing agriculture
CP
LW
BG
Increasing
WT
HS
CP
Discussion
BG
agriculture
Here
manner.
in a dose-dependent
abnormalities
increase with
increasing
agriculture
as males, males with Bidder's organ abnormalities
individuals classified
(Bidder's), or
of individuals with testes and a specific number of female tissues
site. {B) Percentage
of oog?nesis;
the number of female tissues found in toads with testes (0-3) is a meas
of toads with (1) or without
of the intersex condition.
(0) Bidder's organ
(0 Percentage
at each site.
number of gonadal
abnormalities
each collection
site. {D) Maximum
intensity increases along the x-axis from left to right for each graph.
Figure 2. Gonadal
of
{A) Percentage
intersexes at each
or advanced
stage
ure of the seventy
at
abnormalities
Agricultural
2,500
we
show,
for
the first
abnormalities
2,000
males
1,000
800
1,500
600
1,000
400
500
0
t5
200
r^
.?
r^
NAg
Ag Intersex Bidder's Ag
Female Female
male Male
NAg
Male
jfL
Vo
NAg
Ag Intersex Bidder's Ag
male Male
Female Female
NAg
Male
A
Intersex Bidder's Ag
Ag
NAg
male Male
Female Female
Environmental
Health
Perspectives
volume
2008
(B) Testosterone
from agricultural
values.
NAg
Male
con
areas.
1529
HB
McCoy et al.
increase
abnormalities
gonadal
tural intensity.
Gonadal
with
has
agricul
such
abnormalities,
as those
are
to reduce the
likely
reported here,
success
of affected
individuals
reproductive
have
populations
or have
ing
documented
vitellogenin(s)
we
phologic
sentinel
documented
advertising
endocrine
distributed
that an
the
We
disruption.
and therefore
to
provide
large scale
about
the distribution
associated
with
various
can
widely
at
studied
of
land
endocrine
lated via
use
genin(s)
only known
the Bidder's
to occur
after
organ
in males
and
pad
number
are
areas,
again
is compro
are maintained
characteristics
via
are
and
typically modu
in the
produced
gonads
Female
colora
1999).
signaling
sex steroids
color pattern
for estrogenic
is estro
{Hyperolius argus)
of
induction
female
early
has been
as a biomarker
proposed
activity
typical
B. marinus
of female
is due
to dense
concentrations
melanin
at
mammals,
in part,
least
in
estrogens
by
Therefore,
differences
not
does
in B. marinus
is
patterns
increased
there
agriculture,
coloration
dimorphic
associated
to be a clear association
plasma E2 concentrations
testosterone
Plasma
tling.
in all
reduced
agricultural
was
male-typical
B. marinus.
and
between
mot
increased
were
concentrations
testes) as
sexually dimor
testos
be associated with
toads
could
than
(with
so
coloration,
phic coloration
terone
rather
in
concentrations
E2
of P450
Alternatively,
expression
in the skin and local aromatization
aromatase
to estrogens
of androgens
for body color variation.
and Hayes
(Noriega
is
which
castration,
nuptial
from
uses.
of agricultural
diversity
from
sites in this study
agricultural
have
deformities
that clearly
alter
gonadal
function.
Accumulation
of vitello
gonadal
within
of
that naturally
sexually dimor
sexual traits such as skin color,
and
in toads
Sexual
including
Toads
are
2002;
agricultural
that testicular
function
suggesting
in toads from these sites.
mised
informa
important
of abnormalities
forms
found
altered
are
be
also
phic secondary
forearm width,
the effects
of
studying
as visual
serve
"tags"
individual
has experienced
B. marinus
testes
sites have
the agricultural
oocytes
vitellogenic
within
their Bidder's
that
organs,
suggesting
the testes of these toads are not
functioning
to suppress Bidder's
organ oog?nesis.
normally
for
species
traits
These
EDCs.
very
tion
have
of sexually dimorphic
external mor
an ideal
traits that makes
B. marinus
within
the
accumulation
independent
that
amphibian
are declin
to
exposed
pesticides
extinct
Davidson
and
gone
(e.g.,
Furthermore,
et al.
(Brown
that
to suppress
in males
necessary
to the conclusion
led
forearm
Furthermore,
are
number
pad
function
in
could
be
width
responsible
and
androgen
dependent
et al.
(Emerson
mating
nuptial
and
1999;
Qt?
1983). We
in intersex
sex toads
show
these
individuals,
do not build
or
lature
that
associated
are reduced
traits
that
suggesting
inter
muscu
hypertrophied
develop
nuptial
pads
with
normal
levels of
typically
androgens
testes
(Lee and
and properly
functioning
Correales
intersex
toads have
Indeed,
2002).
more
testosterone
reduced
concentrations,
to females
similar
width
and
ers of
NAg
Female
Intersex Bidder's Ag
Ag
Female
male
Male
LW
NAg
Male
Increasing
a
WT
agriculture
HS
?
arm
Thus,
NAgMs.
are biomark
number
pad
status.
androgen
development
toads do not
to
than
nuptial
Because
nuptial
pad
and nonreproductive
is seasonal,
possess
pads, decreased
intersex animals
well-developed
nuptial
nuptial
in
pad development
that they are not
suggests
as
active
as
Moreover,
NAgMs.
are under
width
pads and forearm
selection
(Lee and Correales
2002).
in this study
the abnormalities
found
Thus,
also likely influence
the nature
and action
of
reproductively
3.0
nuptial
sexual
1.5
sexual
selection.
The
occurrence
arise
could
through
of
masculinization
Intersex Bidder's Ag
Ag
NAg
Female Female
male
Male
Intersex
Bidder's
male
Ag
Male
sites.
Error bars
intervals.
1530
volume
masculinized,
intersex
of
individuals
feminization
If females
females.
we would
have
found
traits
wider
this.
female
relatively
complete
tracts and some
combination
(e.g., nuptial
small
arms,
Instead,
normal-sized
intersex
testes,
pads,
testes).
individuals
but
reproduc
of male
decreased
We
varied
were
individu
als with
tive
or
of males
did
mottling,
not find
typically
in ovary
had
size
no
In addition,
of oog?nesis.
stage
females
and oviducts,
but no testes)
(ovaries
or solid coloration.
had nuptial
Females
pads
and
2008
Environmental
Health
Perspectives
Gonadal
not
did
differ
were
males
oration)
in
observe
the
at the
tling
were mottled
B. marinus
at
in color
more
sites.
agricultural
of the
Many
associ
abnormalities
gonadal
in this study are organi
agriculture
the toads'
and remain
throughout
ated with
zational
adult
lives.
Intersex
tinct
ovarian
and
not
reproductive
altered. This
sue
or
after
of
suggesting
that
is fundamentally
to other
that
studies
development
is similar
both ovarian
reported
from
in frogs obtained
have
of dis
that were
tissues
similar,
bilaterally
always
consisted
gonads
testicular
tis
testicular
and
areas
agricultural
to atrazine
tadpoles
exposure
et al. 2003).
These
(Hayes
to
not exclusive
agricultural
are
observations
areas
in other
in study
sites are defined)
how
(e.g.,
design
in the responsiveness
and potential
differences
of subpopulations
for some of
could account
these
An
differences.
concerning
abnormalities
such
environmental
not
question
at which
important
the frequencies
occur
under
remains
conditions.
address
explicitly
examine
populations
we
However,
impact.
Our
this because
with
"normal"
study does
we did not
little
found
the
and fluctua
are
impor
effects
phenotypic
et al. 2006; Guillette
et al. 1995;
(Edwards
a
et al. 2006). Therefore,
Milnes
single mecha
nism of toxicity,
in controlled
identified
labo
tant factors
that determine
to be
is unlikely
alone
ratory settings,
acting
under natural conditions.
Thus, we should not
to
mecha
expect
identify
chemical-specific
nisms of toxicity
in wild
animals. A
occurring
used at our
agricultural
and have well
disruption
studied mechanisms
of toxicity.
Glyphosate
are used at CP and
atrazine
and
(Roundup)
few
the pesticides
endocrine
of
sites cause
BG
and
are known
to
disrupt
Soso
steroidogenesis
et al. 2007).
acute
Glyphosate
disrupts
steroidogenic
regula
across diverse
tory (StAR) protein
expression
taxonomic
the initial
groups, which modulates
et al.
(Oliveira
step
in the
and
2000)
mone
2007;
(Walsh
steroidogenic
pathway
leads to reductions
in steroid
both
et al.
hor
testosterone
production,
including
across several ver
and E2. Research
conducted
tebrate classes has shown that atrazine exposure
can
inhibit
estrogens
(Crain
androgen
production
aromatase
by increasing
et al. 1997; Fan
Environmental
Health
and
increase
transcription
et al. 2007;
Sanderson
Perspectives
have
we
not measured
volume
As
in
expect,
toads are not
might
feral
our
the patterns
consistent
with
entirely
in the
of toxicity
documented
of these well-studied
for either
laboratory
Altered
phenotypes,
including
compounds.
in this study are a
hormone
concentrations,
to several chemi
result of multiple
exposures
over the lifetime
concentrations
cals at various
of the toads.
We
found
in the
anuran
altered
asking
with
and function
B.
amphibian
land
by
dependent
the focus
form
that gonadal
use
in a dose
agricultural
our
fashion. Therefore,
study
literature debate
of the current
if
abnormalities
are
marinus
shifts
from
are associated
gonadal
to a new
line of questions
agriculture
focused on identifying
affected
species, chemi
cal causes,
and developmental,
physiological,
and
ecological
implications
of
exposures.
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is
that testosterone
found
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not.
is
It
but
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among
is still possible
that other
that we
estrogens
human
that
abnormalities
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