Detection of teratogenic substances in acidic mine water samples using the frog embryo teratogenesis assay--Xenopus (FETAX).

TLDR: High concentrations of toxic metals in Tar Creek are identified by water analysis and it is demonstrated, by removing metals via Chelex® 100 ion exchange chromatography, that the observed toxicity and teratogenicity were caused by metal ions.

Abstract: The FETAX (Frog Embryo Teratogenesis Assay — Xenopus) whole embryo bioassay has been developed to screen for environmental substances that cause birth defects. We have used this assay to test its effectiveness in working with actual water samples from the field. Tar Creek is contaminated by discharges from abandoned lead and zinc mines. In addition to high concentrations of zinc, iron and other metals, water samples are routinely low in pH and oxygen content. The pH values of three Tar Creek sample sites were below the established tolerance limits of the embryos. Therefore, one group of samples had no pH adjustment while a second group had the pH adjusted to 7.0. Two of the four sites contained agents that reduced embryonic growth and caused high rates of mortality. A third site contained teratogenic substances. We have determined that metal content is responsible, along with the low pH, for the biologic effects. We have identified high concentrations of toxic metals in Tar Creek by water analysis and were able to demonstrate, by removing metals via Chelex® 100 ion exchange chromatography, that the observed toxicity and teratogenicity were caused by metal ions. We have concluded that FETAX is an excellent test for complex mixtures. Physicochemical parameters, such as pH, oxygen and metals content, can be altered and the effect of these changes on toxicity and teratogenicity determined using FETAX. The interaction of toxic substances and low pH are important when considering embryo survival and development.

Figures

Table 3. cont. Horse Pasture

Table 3. cont. Horse Pasture

Table 2. Con~entration Ranges of Divalent Metal Ions for Each Sample Site •

Figure 1. Tar Creek area map and locations of study sites. Tar and Treece-Tar Creek and Treece, OK Sl8 T29N R23E OWRB# 4-East of Tar and Lytle Creek junction S29 T29N R23E Horse Pasture-South of Commerce, OK S7 T28N R23E Tar and Miami-N.E.O. A&M and Tar Creek bridge S30 T28N R23E

Table 3. cont. Horse Pasture

Full text

DETECTION
OF
TERATOGENIC
SUBSTANCES
IN
ACIDIC
MINE
WATER
SAMPLES
USING
THE
FROG
EMBRYO
TERATOGENESIS
ASSAY-XENOPUS
(FETAX)
By
DOUGLAS
ALAN
DAWSON
!I
Bachelor
of
Arts
Transylvania
University
Lexington,
Kentucky
1980
Submitted
to
the
Faculty
of
the
Graduate
College
of
the
Oklahoma
State
University
in
partial
fulfillment
of
the
requirements
for
the
Degree
of
Master
of
Science
May,
1985

DETECTION
OF
TERATOGENIC
SUBSTANCES
ACIDIC
MINE
WATER
SAMPLES
USING
THE
FROG
EMBRYO
TERATOGENESIS
ASSAY-XENOPUS
(FETAX)
Thesis
Approved:
~-yJ~
~~f
the
Graduate
College
1216263
'
i i

PREFACE
The Frog Embryo
Teratogenesis
Assay-Xenopus
(FETAX)
has been
developed
to
detect
teratogenicity
of
pure
compounds and complex
mixtures.
The
application
of
this
system
as a
quick
screen
to
determine
possible
teratogenicity
of
test
substances,
allows
for
prioritization
of
those
substances
for
further
testing
in
established
mammalian
systems.
FETAX
provides
data
on
malformation,
mortality
and
inhibition
of
growth and
development
that
can be
statistically
analyzed
to
determine
the
teratogenic
risk
of
the
test
compound.
This
study
determined
the
presence
of
teratogenic
agents
in
acidic
mine
water
samples
from
the
Tar
Creek
area
of
Oklahoma.
The
results
obtained
were
another
step
in
the
validation
of
FETAX
as a
useful
method
of
determining
agents
that
cause
birth
defects.
I would
like
to
thank
the
people
at
Oklahoma
State
University
who
assisted
me
in
the
progress
and
completion
of
my
program.
I
particularly
wish
to
express
my
gratitude
to
my
major
adviser,
Dr. John
A.
Bantle,
for
his
involvement
and
assistance
in
my
course
of
study.
i i i

I wish
to
thank
my
other
committee
members, Dr.
Calvin
G.
Beames,
Jr.
and Dr. S.
L.
Burks
for
their
assistance
throughout
my
work.
Special
thanks
are
due
to
the
staff
of
the
Water
Quality
Research
Lab,
under
the
direction
of
Dr.
Burks,
especially,
Curt
McCormick
for
his
collection
of
the
water
samples
and
information
on
the
water
chemistry
of
the
area
and
Sarah
Schatz
for
her
time
and
effort
involved
in
the
water
sample
analysis.
I
am
also
grateful
to
those
who
provided
technical
assistance
and animal
care
during
the
study:
Douglas
Bailey,
Mitchell
Edmondson,
Melanie
Lamb,
Dana
Ramsey,
Patricia
Simpson, Marianne Swanner and
Clementine
Yap.
Final
thanks
are
extended
to
the
University
Center
for
Water
Research
at
Oklahoma
State
University
and
to
the
March
of
Dimes
Birth
Defects
Foundation
who
supported
this
work.
i v