Marine Ecology
Research Highlights
National Marine Water Quality Laboratory
P. O. Box 277 West Kingston, R. I. 02892
Vol. 1., No. 1 - February 1973
Here -in this little bay
Full of tumultuous life and great repose
Where, twice a day,
The purposeless glad ocean comes and goes,
Under high cliffs and far from the huge town
I sit me down.
For want of me the world's course will not fail.
When all its work is done the lie shall rot.
The truth is great, and shall prevail
When none cares whether it prevail or not.
Coventry Patmore's poetic expression of the aesthetic and
spiritual inspiration to be dravm from the estuarine environment
is indeed a beautiful thought—but truth cannot prevail on its own
merits alone, given the already serious problems of domestic and
industrial uses of inshore waters. Although the poet may look
ahead through eons, our vision has to be restricted to the present
and the immediate future—for our tasks deal daily with engineering,
ecological, and legal methods to assure the wisest and best uses of
our bays, estuaries, and coastal waters for all the days to come.
Our main thrust is in ecological research, but we also feel the
obligation to participate with EPA's engineering and legal staffs
in applying our findings to the permits and enforcement activities
through which the future of our marine resources is decided. To
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show you how this is done, we shall trace one research effort from
its beginning in our West Kingston laboratory through its develop-
ment, testing, refinement, dissemination, and applications in
Massachusetts, Florida, the Chesapeake, New Jersey and Texas.
An Associate Laboratory of the National Environmental Research Center * Corval'is
200 S.W. 35th Street. Corvallis, Or. 97330
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SEARCH FOR AN ENVPOTNT :
Microscopic plants are the grass of the oceans——forming the
base of the marine food web. Their species exist in great diversity
in healthy bays and estuaries, but under great stress of pollution
a few nuisance species bloom as great numbers of tiny individuals——
taking over the bay to the detriment of all other creatures. At
NMWQL, we study these microalgae to determine their requirements
and tolerances so that EPA water quality standards may account for
the needs of all members of art ecosystem. Because these plants are
microscopic, it is difficult to tell their state of health under
experimentally varied conditions——so we have tried a number of in-
direct chemical methods used commonly to evaluate their life, death,
or physiological condition.
Some methods work well in the labora tory but not always in the
field——such as measuring the uptake of radioactive carbon. Some,
such as measuring the ratio of chlorophyll a to phaeopigments (a
degradation product of photosynthetic pigments), are insufficiently
sensitive to rapid changes in condition. Following population
abundance and distribution by direct microscopic observation is
still best, but is slow and requires an expert botanist——a luxury
that many laboratories cannot afford. We needed something fast,
technically simple, easily taught, reasonably inexpensive, and highly
reliable to use as an endpoint in laboratory experiments and field
evaluations of the condition of microscopic plants (and animals) in
nearshore marine waters.
EUREKA - ALMOST .
Stan Hegre saw it first——a paper describing the measurement of
adenosine triphosphate (ATP) to determine the living biomass in a
volume of water. As a biochemist, he realized that a freshly killed
cell loses its ATP in seconds, but a living cell maintains a rela-
tively constant level, because ATP is a universal metabolite inter-
mediate in the conversion of energy to food and back to energy.
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The analysis is quick and easy——common in medical research
laboratories——but had never been applied to the sorts of marine
questions that we were asking. Dr. Hegre saw not only that it
would work in the laboratory, but that we had a pressing and
innuediate field application at hand.
TUE KEYSTOWE COPS MEET TUE PERILS OF PAULiNE .
The field application at hand was that our Division of
Enforcement needed to know the effects upon the marine plankton
in Biscayne Bay at the Turkey Point power plant, and they needed
the answer quickly. We were in court, and our case had not in-
cluded studies of the plankton. Opposition witnesses contended,
based only upon microscopic observation, that the plankton was
not being harmed. Our legal staff believed that damage to the
plankton could be shown.
NMWQL was called upon at this point. Drs. Hegre, Gonzalez,
Steele, Gentile, Davey, and Prager joined the Regional personnel
already working at Turkey Point and initiated plankton studies,
comparing the waters at the intake, just beyond the plant’s con-
densers, and at the point of discharge into Biscayne Bay.
We were on our first deadlined field support study and must
have looked, to our cooler colleagues from Region IV, like the
Keystone Cops——running about in search of a workable method and
equipment to work it with. Radioactive carbon uptake had failed
due to too much carbonate sand in the water. Plant pigment changes
told us nothing about whether the heat, the chlorine, the duration
of exposure, or the pumping was causing harm to the plankton popu-
lation. It was then that Dr. Hegre thought of the ATP technique.
Would it work? Could we untie our lawyers from the railroad tracks
in time to get them to court with the needed evidence?
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ATP TO TUE RESCUE .
We sampled the three stations, each time in triplicate, filter-
ing 200 ml portions of pooled Van Dorn collecting bottle samples
through membrane filters minutes after the samples were taken. Im-
mediate extraction in boiling TRIS buffer solution followed and then
storage of the extract in the freezer completed the field portion of
each day’s ATP work on whole water samples. While this was going on
we separated out the zooplankton, cataloged numbers of living and
dead, and analyzed their ATP content by the same extraction procedure.
Extracts could be stored frozen, awaiting a let—up in the
hurried sampling schedule, when they were analyzed on a luminescence
biometer borrowed from Dr. Weber at the Cincinnati Analytical
Quality Control Laboratory. Data piled up for two months, and
looked good——good agreement of replicates——clear differences among
stations——rapid responses to changes in the plankton with passage
through the plantts condensers.
It worked. Indeed, the plankton were being harmed by summer
passage through the plant. Seventy—five percent of the zooplankton
were killed just in passing through, and by the time the cooling
waters had reached Biscayne Bay, some 4 hours and 2 miles later,
another 10% were lost. Half of the microscopic algae had been
lost in the passage. EPA and Department of Justice Attorneys
were able to use this information in the settlement that followed.
Pauline had been saved just in time.
WAITER, THERE’S A COPEPOV IN MY SOUP !
We were lucky that Turkey Point had been an oversimplified
field sampling situation in which our mobile laboratory was very
near the sampling stations. We realized later, planning to sample
Galveston Bay for the U.S. vs. Houston Lighting and Power Co . case,
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that if we carried our water samples for any distance, the elapsed
time would be great enough to show effects of that increased ex-
posure and therefore spoil the ATP analyses. This indeed was the
marine equivalent of a fly In our soup. We had to develop a method
to treat the samples immediately, In the field, so that the ATP
would remain stable until it could be extracted and analyzed. Sue
Cheer had an Idea.
Liquid nitrogen would freeze the filter pact at —196°C to fix
the sample Instantly. The problem was devising hardware to handle
and store liquid N in the field. We would be sampling from bouncy
small boats and helicopters, and liquid N is rather nasty stuff to
have sloshing about. While Sue Cheer, Pete Rogerson and Jack
Gentile were testing the liquid N as a fixative against the standard
no—fixative method, they also designed and built styrofoam insulated
boxes to hold the liquid N filter pad containers.
Pete Rogerson also found that ordinary vacuum bottles from our
lunch boxes could be vented through the plastic cap to serve as
field storage vessels from which the sample containers could be re-
plenished as needed. Since neither small boats nor helicopters have
a vacuum pump, Sue Cheer worked out a way to filter the samples by
pressure using a 50 ml hypodermic syringe and Swinney filter holder.
One field test, in a small boat off Point Judith (known in the
1600’s as Point Judas) on a particularly seasickey day, proved to
our satisfaction that the method would work as well in the field as
the sampling team would.
AVVAUTAGE.S VS. LIM1TATIO IS OF TI-fE AlT ANALYSTS .
Remember that no indirect chemical assessment of plankton
abundance and distribution can be used by itself to form an environ-
mental value judgment——because none yet devised can distinguish good
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species from undesirable ones. Do not be misled by implied, value
statements referring to “increased productivity” or “enriched
waters” — — or attempts to have you believe that “the bodies of
killed plankton merely reenter the nutrient cycle as dissolved
chemicals without effect on the balance of the indigenous popula-
tion”.
Abundance and distribution of species must be determined by
direct observation. Indirect chemical methods indicate their con-
dition and permit you to make microscopic observations less
frequently, saving expert manpower by spot checking abundance and
distribution. Different chemical evaluations provide different
information about the biota——and these may add to a species by
species analysis of environmental condition——but can never replace
it.
In the final analysis, a healthy plankton population consists
of a large number of desirable food species——each represented by a
few individuals. Any trend toward a population dominated by few
species, each in large numbers——or a predominance of inedible or
toxic species——represents a decrease in water quality.
ATP analysis is the fastest chemical assessment of plankton
condition. Plant pigments react more slowly to change. Even the
ratio of chlorophyll a to phaeopiginents, which also gives an index
of phytoplankton (but not zooplankton) condition, requires many
minutes to reflect a kill. l 4 C uptake is almost as fast as ATP
but is applicable only to phytoplankton and cannot be made to work
reliably if there are large amounts of inorganic carbon in the
water from nonliving sources. It is a useful open ocean method
which becomes excessively difficult in certain estuarine environ-
ments. Neither electronic particle counting (and sizing) nor
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analysis for total, particulate, or dissolved organic carbon are
capable of distinguIshing living from dead or detrital materials.
Again we must caution never to use any indirect chemical assessment
of the plankton without some degree of direct observation.
In terms of capital cost, plant pigment analysis is least ex-
pensive (2-3$K), followed by 1 - 4 C uptake and ATP which are comparable
(5—6$K), and electronic particle analysis (8—20$K). However, a
laboratory having a liquid scintillation counter can use It for ATP
analysis and so avoid the cost of special instrumentation (i.e., the
super—sensitive recording light meter that measures the flash given
off when ATP is added to a light emitting enzyme—substrate mixture
extracted from the south end of a northbound firefly).
GOOV NEWS TRAVELS FAST .
Even before Cheer, Rogerson, Gentile, and Hegre have published
the ATP method application, It is being put to use in a number of
power plant entrainment studies. NMWQL reports on Turkey Point and
Cedar Bayou studies proved its applicability in the Texas Gulf and
on Florida’s East Coast. Florida Power Corporation’s consultants
are using ATP analyses on the Gulf Coast at the Crystal River and
Anclote sites. Raytheon is using it at the B.L. England plant in
Little Egg Harbor, N.J., and the Massachusetts State laboratory at
Cat Cove Is initiating an ATP study of Salem Harbor’s power plant.
VO CALL US, WE WON’T CALL YOU .
Our commercial phone number is 401/789—9751 and FTS users may
direct—dial 401/528—4371. Drs. Sue Cheer and Pete Rogerson can tell
you about comparisons between our method modifications and the orig-
inal technique. Drs. Jack Gentile and Stan Hegre can tell you about
comparisons between ATP analysis and other indirect assessments of
plankton condition, abundance, and distribution, as well as the use
of ATP concentration as a bioassay end—point. Requests for training
or technical assistance should be routed through Dr. Jan C. Prager.
We’ll be glad to hear from you. Ivory towers get cold and lonesome.
— — — — J.c.P.
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