A Dirty Water Biologist’s
View
Of
Water Quality
William T. Waller (Tom) University of North Texas
Institute of Applied Science
and
Department of Biology
Toxicologists are guided by principles
Principle 1
In terms of measuring chemicals – you
only find what you are looking for and
you only find it if it is present in
sufficient quantity to be detected by
the method you are using to measure
it.
Pecan Creek Above WWTP
Pecan Creek at WWTP Discharge
Activated Sludge Aerobic Digestion
Pecan Creek
Aerobic Digesters
Final Clarifiers
Dynodirt
Primary Clarifiers
Landfill
1.
Coagulation removes dirt and other
particles. Alum and other chemicals
are added to form sticky particles.
The combined weight of the
particles makes them settle.
2.
The water passes through filters,
some are made of layers of sand,
gravel and charcoal that help
remove even smaller particles.
3.
A small amount of chlorine is added
(or some other disinfection method
is used kill any bacteria or
microorganisms that might be in the
water.
4.
The water then moves through the
distribution system.
1
2
3
4
CH3
C
CH3
CH
CH3
N
CH
C
C
N
OC2H5
O
S
P
OC2H5
Diazinon
O,O-Diethyl O-(2isopropyl-4-methyl-6-pyrimidinyl) phosphorothioate
acetylcholinesterase inhibitor
insecticide, nematicide
Use: For soil insects and pests of fruits, vegetables, tobacco, forage, field
crops, range, pasture, grasslands, ornamentals. For cockroaches and other
household insects including grubs, nematodes in turf, seed treatment and fly
control.
How many football field sized containers (100 yds x 50
yds x 3 yds) three yards deep would it take to dilute the
diazinon in a quart container of 25% diazinon to the
recommended concentration of 80 ng/L?
Conversions
To convert from
To
Multiply by
gm
mg
ug
ml
cc
qt
liters
gallons
ft3
mg
ug
ng
gm
ml
ml
gallons
ft3
yd3
1,000
1,000
1,000
1
1
946.3
0.26417205
0.13368
3.7037x10-2
Diazinon – “Safe” concentration for aquatic life is 80 ng/l,
although newer research indicates that “safe” might be as
low as 8 ng/l.
The concentration of in 25% daizinon is 25% by weight. One
liter contains _______ ml, and weighs _________ gms.
One quart contains _______ ml, therefore a quart weighs
_______?
If one quart weighs _______ gms and 25% of that is
diazinon then one quart contains ________ gms of diazinon.
________ gm of diazinon = __________ mg of diazinon
________ mg of diazinon = __________ ug of diazinon
________ ug of diazinon = __________ ng of diazinon
How many liters of water are needed to be added to the
solution containing _______ ng of diazinon to reduce it to
80 ng/l?
___________ ng = 80 ng/l
x
x = ____________ liters
To convert liters to gallons multiply by _______ = ______
To convert gallons to ft3 multiply by ________ = _______
One yd3 contains 27 ft3 to convert ft3 to yd3 divide by 27 =
___________
How many containers 100 yds long by 50 yds wide by 3 yds
deep does it take to dilute the amount of diazinon in a quart
of 25% diazinon to 80 ng/L 258
If it turns out that the “safe” concentration is 8 ng/l, how
many boxes would it take? 2580
Municipal Separate Storm Sewer Systems – MS4s
Non-point source water does not, for the most part go
through any treatment before it reaches a receiving
system. For that reason anything that you put on your
land, or dump down a storm drain goes into your
drinking water source without treatment.
So, is diazinon in our drinking water? I don’t know we
don’t measure for it and you only find what you are
looking for!
Principle 2
The dose makes the poison! Paracelsus
(1493-1541) is credited with this
dictum when he wrote, “All substances
are poisons, there is none which is not
a poison. The right dose differentiates
a poison from a remedy.” In aquatic
toxicology we are concerned with
exposure which we can define as the
magnitude, duration and frequency
with which an organism interacts with
biologically available toxicants.
ü On average, a penny minted
since 1983 contains
approximately 59,500 ug of
copper and 2,420,000 ug of
zinc.
ü Needless to say that much
copper and zinc is sufficient to
cause significant damage to
fish living in most fountains
and coy pools.
ü Yet when the penny (pennies)
are tossed into the fountains
of pools the fish seem to do
just fine. How can this be?
Principle 3
“No instrument has yet been devised that can
measure toxicity. Chemical concentrations can
be measured with an instrument but only living
material can be used to measure toxicity.”
John Cairns, Jr. and D.I. Mount. 1990.
Environmental Science and Technology
The point is, that living material i.e., biological
monitors (biomonitors) need to be used to
measure toxicity. Biomonitors are the only
things that integrate the totality of their
environment.
However, the biomonitors do not, for the
most part, tell you directly what is causing the
toxicity. Biomonitors tell you only that
toxicity is or has occurred. Therefore, both
biological and physical/chemical measures
need to be used in biomonitoring
Canary
in the
coal min
e
s
r
e
t
Tas
Principle 4
The Precautionary Principle-This principle is
applied more in Europe than it is in the United
States. It states that in the face of a high
level of uncertainty act conservatively. In
practice it would be invoked when there was,
for example, a chemical that was going to be
released into commerce but not all the testing
on the chemicals safety had been done yet.
The conservative action might be to wait until
all the data are in before releasing the
chemical. Sometimes we don’t do this.
Principle 5
Duck Tape
Duct Tape
Montague
Cooke
Grayson
Collin
Denton
Wise
Tarrant
Dallas
UNT Lab
BioSensor
Composite
Sampler
BioSensor
BioSensor
Proximity Sensor
Clam
Nylon Bolt
Stainless Steel Target
Plexiglas Frames
Nylon Nut
Ceriodaphnia dubia
( water flea)
Pimephales promelas
fathead minnow
pH
DO
Temp
Conductivity
Fluorescence
Depth
Turbidity
Multiprobe
NEMA4
Enclosure
NEMA 4
Enclosure
Computer
Charge
Controller
Battery
A/D Board
Charge
Controller
Computer
Modem
Battery
Nature has this marvelous way of responding
unexpectedly to our insults.
Homeostasis
Two systems control all physiological processes
The nervous system exerts point-to-point
control through nerves, similar to sending
conventional messages by telephone (hard wired).
The endocrine system broadcasts its hormonal
messages to essentially all cells by secretion into
blood and extracellular fluid. Like a radio broadcast
it requires a receiver (cells must have a receptor for
the hormone being broadcast).
These two system often act together
Endocrinology is the study of hormones, their
receptors and the intracellular signaling pathways
they invoke.
Distinct endocrine organs are scattered throughout
the body.
Hypothalamus
Pituitary
Parathyroid glands
Testes
Thyroid glands
Pancreas
Adrenal glands
Ovaries
In addition to the classical endocrine organs,
many other cells secrete hormones, through what is
sometimes called the “diffuse” endocrine system.
All pathophysiological events are influenced
by the endocrine milieu
All “large” physiologic effects are mediated
by multiple hormones acting in concert
There are many hormones known and little
doubt that others remain to be discovered.
Two important terms are used to refer to molecules that
bind to the hormone-binding sites of receptors:
Agonists are molecules that bind the receptor and induce
all the post-receptor events that lead to a biologic event.
In other words they act like the “normal” hormone.
Antagonists are molecules that bind the receptor and
block binding of the agonist, but fail to trigger
intracellular signaling events. (Antagonists are like
certain types of bureaucrats – they don’t themselves
perform useful work, but block the activities of those
that do have the capacity to contribute.) Hormone
antagonists are widely used as drugs.
Normal response
ng
i
Tim
Timing
Abnormal Response
No Response
Dormant receptor
Receptor Blocked
turned on by mimic
Normal receptor over
stimulated by mimic
g
n
i
m
Ti
Death rate < Birth rate
Ova
Sperm
Individual
Born
Lives
Reproduces
Dies
?
Population
Increases
Population
Decreases
Population Persists
Total Fertility
2.1
Death rate > Birth rate
Endocrine System regulates biological processes
• Growth and function of reproductive system (androgen
[testosterone], estrogen [estradiol], and related compounds
from gonads)
• Control of blood sugar (pancreatic insulin)
• Regulation of metabolism (adrenal cortisol and thyroid
thyroxin)
• Development of nervous system including brain (estrogen and
thyroid hormones)
• Overall development from conception to old age = homeostasis
Estrogen Mimics
• Intended disruptors
– DES (diethylstilbestrol)
– ethynylestradiol
– phytoestrogens
Estrogen Mimics
Estradiol
Intended disruptors
-C=CH
Ethynylestradiol
DES
Estrogen Mimics : promiscuous receptor
Estradiol
Unintended disruptors
-C9H19
Bisphenol A
DDT
DEHP
Nonylphenol
Specific Case Example
Fish Response to Wastewater Effluent
Estrogenicity :
Hypothesis: Male fish exposed to effluent
from the City of Denton Wastewater
Treatment Plant would not develop
vitellogenin.
Vitellogenin Biomarker : fish model
Endogenous / Exogenous Estrogens
Vitellogenin
Liver
Gonad
Vitellogenin – egg yolk precursor produced in the liver
Pilot Study Methods
• Adult male fathead minnows (n=5/exposure)
• 1-3 Weeks of exposure to 100% WWTP
• RHW control in lab
• Plasma VTG content via ELISA
• Likelihood Ratio Test
– ND left censored at 3,000 ng/ml
Pilot Study Results
Mean plasma vitellogenin concentrations (ng/ml).
30000
*
25000
*
20000
15000
10000
5000
0
Control
1 Week
2 Weeks
3 Weeks
* Statistical significance from the control (a=0.05).
Comparison to Similar Studies
Mean plasma vitellogenin concentrations (ng/ml) in fish
exposed to wastewater effluent receiving systems.
50000
45000
40000
35000
30000
25000
20000
15000
10000
5000
0
Background
Exposed
TX FHM
UK TRT
UK FLO
MI FHM
Wetland Exposure Design
2
4
Out
3
1
In
Fish Measurements
• Vitellogenin Content of Plasma
• GSI – Gonado Somatic Index testes wt/total
body wt x 100
• HIS – Hepato Somatic Index liver wt/total body
wt x 100
• Hematocrit (%packed blood cells)
• Secondary Sex Characters (turbercles, fat pad, stripes)
• Length
• Weight
• Gonadal Differentiation
Chemical Analysis
-C=CH
Estradiol
Ethynylestradiol
- C9H19
Bisphenol A
DDT
DEHP
Nonylphenol
Wetland Characterization
• vegetation types and density
• depth and width of channels
• retention time as estimated by input flow
• relative to chemical constituents & fish response
Endocrine Disruption in Wildlife
• related to reproduction and sexual development
•
reptile development : alligators (Alligator
mississippiensis) and turtles (Trachemys scripta) which
showed altered sexual development and limited
reproductive potential resulting in population declines
(Guillette, et al., 1995, Crews et al., 1995)
•
birds suffered breeding and embryonic development
effects resulting in reproductive failure across
numerous species (Janz and Bellward, 1997; Fry, 1995;
Henshel, et al., 1995; Bowerman, et al.,1995; White and
Hoffman, 1995; Henny, et al., 1995)
Endocrine Disruption in Wildlife
Decline in amphibian populations worldwide (Hayes, et al. 1997;
Carey and Bryant, 1995)
Intersex fishes (Raloff, 1995; Sumpter, 1995) prompted a reinvestigation of the use of estrogen and synthetic estrogens in
commercial fisheries (Piferrer and Donaldson, 1989; Bye and
Lincoln, 1986; Pottinger, 1986). Further study showed the
capability of xenoestrogens to interact not only with female
estrogen receptors but also with naturally dormant receptors in
male fish
Endocrine Disruption in Wildlife
• mass mortalities of beluga whales (Delphinopterus
leucas) and bottlenose dolphins (Tursiops truncatus) changes in the thyroid gland and the adrenal cortex
which resulted in immunosuppression and
susceptibility to otherwise mild pathogens (De Guise, et
al, 1995; Lahvis, et al., 1995)
• decline of the Florida panther (Felis concolor coryl)
population has also been linked to EEDs with
reproductive, immune and a host of other stereotypical
EEDs effects (Facemire, et al., 1995)
•
follicular cell adenoma in mice (Greenman, et al.,
1990), liver tumors in golden hamsters (Metzler, et al.,
1990)
Endocrine Disruption in Wildlife
Malformations at every target of estrogens in the mammalian
body (including humans) from the mammary glands themselves
to the testes to the skeleton (Dorgan, et al., 1997; Majdic, et al.,
1997; Hajek et al., 1997; Migliaccio, et al., 1995; Iguchi, et al.,
1995; Gorski and Hou, 1995; Rier, et al., 1995)
More pronounced if the developing fetus is exposed across the
fetal placental barrier (Guillette, et al., 1995; Jones and Hajek,
1995; Whyatt and Perera, 1995; Guo, et al., 1995).
Endocrine Disruption in Humans
• Is there a link between wildlife effects and
human impacts ?
• Relationship between environmental estrogens
and human disease, reduced fertility and
associated link to cancer?
More Info
• Our Stolen Future, Colborn, Dumanoski, Myers, 1996.
• Generations at Risk, Reproductive Health and the
Environment, Schettler, Solomon, Valenti, Huddle, 1999.
• http://website.lineone.net/~mwarhurst/chemicals.html
• http://www.tmc.tulane.edu/ecme/eehome/
• http://www.epa.gov/scipoly/oscpendo/index.htm
Acknowledgements
• Students: Dr. Richard Guinn, Dr. H. Joel Allen, Dr. Jon
Hemming, Dr. Tim Knight, Jr., Dr. Jim Lazorchak, Mr. Phil K.
Turner, Tony Clyde, Patricia Lambert plus many others
• Colleagues: Dr. Ken Dickson, Dr. James H. Kennedy, Dr.
Larry Ammann, Dr. Miguel Acevedo, Dr. Steve Nolen
• US EPA, City of Denton, Texas Water Research Institute,
U.S. Army Corps of Engineers, Tulsa District