EPA
United States
Environmental
Protection Agency
Office of
Research and
Development
Qff tc& ol Energy,
Minerals and Industry
Washington. D.C. 20460
EPA-600/7-77-111

October 1977
           PROGRAM REVIEW
           PROCEEDINGS OF:
           ENVIRONMENTAL  EFFECTS OF
           ENERGY RELATED ACTIVITIES
           ON MARINE/ESTURINE
           ECOSYSTEMS

           Interagency
           Energy-Environment
           Research and Development
           Program Report

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                RESEARCH REPORTING SERIES

Research reports of the Office of Research and Development, U.S. Environmental
Protection Agency, have been grouped into nine series. These nine broad cate-
gories were established to facilitate further development and application of en-
vironmental technology.  Elimination of traditional grouping was consciously
planned to foster technology transfer and a maximum interface in related fields.
The nine series  are:

      1   Environmental  Health Effects Research
      2,  Environmental  Protection Technology
      3.  Ecological Research
      4   Environmental  Monitoring
      5.  Socioeconomic Environmental Studies
      6.  Scientific and Technical Assessment Reports (STAR)
      7   Interagency  Energy-Environment Research and Development
      8,  "Special" Reports
      9.  Miscellaneous Reports

This report has  been assigned to the INTERAGENCY ENERGY-ENVIRONMENT
RESEARCH AND DEVELOPMENT series. Reports in this series result from the
effort funded  under the 17-agency Federal  Energy/Environment Research and
Development Program. These studies relate to EPA's mission to protect the public
health and welfare from adverse effects of pollutants associated with energy sys-
tems. The goal  of the  Program is to assure the rapid development of domestic
energy supplies in an environmentally-compatible manner by providing the nec-
essary environmental data and control technology. Investigations include analy-
ses of the transport of  energy-related pollutants and their health and ecological
effects; assessments of, and  development of, control technologies for energy
systems; and integrated assessments of a wide range of energy-related environ-
mental  issues.
 This document is available to the public through the National Technical Informa-
 tion Service, Springfield, Virginia 22161.

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                                       OEMI-77-025
                                       October  1977
         PROGRAM REVIEW PROCEEDINGS OF:
         ENVIRONMENTAL EFFECTS OF ENERGY RELATED
         ACTIVITIES ON MARINE/ESTURINE ECOSYTEMS
                  compiled by

       Clinton Hall and Walter Preston
              Project Officer
              Walter Preston
Office of Energy, Minerals, and Industry
   Office of Research and Development
         Washington, D.C.  20460
Office of Energy, Minerals, and Industry
   Office of Research and Development
  U.S. Environmental Protection Agency
        Washington, D.C.  20460

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                                  FOREWORD
     Although the Federal Government has been the major sponsor of energy
research and development since the early 1950's, it has been only recently
that energy/environment R&D has been of major public concern.  Since fiscal
year 1975, the Environmental Protection Agency has had the lead responsibility
for the planning, coordination, and implementation of the Federal Interagency
Energy/Environment R&D Program.

     This Program, funded at approximately $100 million per year and
participated in by seventeen Federal departments and agencies, is based on
the recommendations of two interagency working groups commissioned by the
Council on Environmental Quality and the Office of Management and Budget
in 1974.  The "Report of the Interagency Working Group on Health and
Environmental Effects of Energy Use," completed in November 1974, recommended
an integrated environmental sciences research program addressing needs in
five major areas:

           o  Pollutant Characterization and Monitoring
           o  Environmental Transport Processes
           o  Health Effects
           o  Ecological Effects
           o  Integrated Assessment

     The program  is now in its fourth year.  We are committed to keeping
the Interagency Energy/Environment Program objective oriented.  Accordingly,
a series of program review and planning sessions have been scheduled for each
subprogram area both to examine the science being conducted at the specific
project level and also to draw on the knowledge and instincts of the
participating scientists and program managers to forecast what the future
research needs will be.  It is hoped that in this manner we can adequately
anticipate and provide the kind of information that will ensure sound
environmental consideration in future energy policy and decision-making.

     The following compilation represents the proceedings from the review
session for the marine and estuarine portion of the interagency program 
cross-cutting the pollutant characterization and monitoring, environmental
transport processes, and ecological effects subareas.
                                         Clinton W. Hall, Director
                                         Energy Coordination Staff
                                         U.S. Environmental Protection
                                                  Agency

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                           TABLE OF  CONTENTS
                                                                  Page
SUBLETHAL EFFECTS OF PETROLEUM  ON MACOMA BALTHICA                  1
(M. S. Stekoll, L. E. Clement,  and  D.  G.  Shaw)

THE BENTHIC COMMUNITY OF A NATURAL  OIL SEEP IN  THE SANTA
   BARBARA CHANNEL                                                 19
(Robert B. Spies, Paul H. Davis, and Thomas M.  Distler)

ACUTE AND CHRONIC STUDIES WITH  WATERFOWL EXPOSED TO PETROLEUM
   HYDROCARBONS                                                    35
(Michael P. Dieter)

ACCUMULATION AND BIOLOGICAL  EFFECTS OF COPPER AND CADMIUM ON
   MARINE SHELLFISH                                                45
(David W. Engel and Bruce A.  Fowler)

ECOLOGICAL CHARACTERIZATION:  AN AID TO DECISION MAKING            57
(Howard D. Tait)

AN ECOLOGICAL CHARACTERIZATION  OF THE  CHENIER PLAIN OF
   SOUTHWESTERN LOUISIANA AND SOUTHEASTERN  TEXAS:   OBJECTIVES
   AND METHODOLOGIES                                               65
(Janie Harris)

THE CONSTRUCTION OF A CONCEPTUAL MODEL OF THE CHENIER PLAIN
   COASTAL ECOSYSTEM IN  TEXAS AND LOUISIANA                       75
 (L. M. Bahr,  Jr., J. W.  Day,  Jr., T. Gayle, J.  G. Gosselink,
C. S. Hopkinson, D. Stellar,  and J. Harris)

FATE AND EFFECTS OF CONTAMINANTS IN MARINE  ECOSYSTEMS AND
   ORGANISMS                                                       93
(Douglas A. Wolfe)

LABORATORY AND FIELD STUDIES  ON THE LONG-TERM EFFECTS OF
   PETROLEUM HYDROCARBONS ON  BENTHIC MARINE INVERTEBRATES        107
(J. W. Anderson, R. Riley, B. M. Bean, J. W.  Blaylock, and
S. L. Kiesser)

ENVIRONMENTAL ASSESSMENT OF  NORTHERN PUGET  SOUND AND THE
   STRAIT OF JUAN DE FUCA                                         133
(Howard Harris)

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TABLE OF CONTENTS  (Continued)
                                                                 Paqe
ENVIRONMENTAL ASSESSMENT OF AN OFFSHORE OIL FIELD:
   A PROGRESS REPORT                                              143
(Richard J. Berry and Ernest D. McRae)

A STANDARD PROGRAM FOR ENVIRONMENTAL  IMPACT ASSESSMENT:
   PHASE I  ICTHYOPLANKTON SAMPLING                            153
(Martin A. M. Hyman, Ernesto Lorda, and Saul B.  Saila)

EFFECTS OF CHEMICAL USE IN OFFSHORE WELL-DRILLING
   OPERATIONS                                                     161
(Norman L. Richards)

THE CHEMISTRY AND ECOLOGICAL EFFECTS  OF CHLORINATION OF
   SEAWATER:  A SUMMARY OF EPA RESEARCH PROJECTS                 175
(W. P. Davis, D. P. Middaugh, J. H. Carpenter, G. R. Helz,
and M. H. Roberts, Jr.)

POWER PLANT COOLING WATER CHLORINATION IN NORTHERN
   CALIFORNIA                                                     187
(S. Hergott, D. Jenkins, and J. F. Thomas)

THE NBS PROGRAM FOR STANDARDS FOR TRACE ORGANIC  ANALYSIS
   IN THE MARINE ENVIRONMENT                                      227
(H. S. Hertz, S. N. Chesler, W. E. May, S. A. Wise,
L. R. Hilpert, J. Brown, A. Fatiadi,  and F. Guenther)

THE NBS PROGRAM FOR STANDARDS FOR TRACE INORGANIC
   ANALYSIS IN THE MARINE ENVIRONMENT                            243
(I. L. Barnes, H. L. Rook, and T. J.  Murphy)

STANDARDIZATION AND INTERCALIBRATION  TECHNIQUES  FOR
   MARINE MONITORING                                              259
(Michael A. Basileo)

DEVELOPMENT OF OCEAN OIL SPILL CONCENTRATION AND TRAJECTORY
   FORECAST METHODS                                               271
(Celso S. Barrientos)

THE UNDERWAY WATER SAMPLING SYSTEM                                283
(C. D. Kearse, R. New, and D. Enabnit)

SHIPBOARD ENVIRONMENTAL DATA ACQUISITION SYSTEM                   301
(Douglas W. Downen and Richard R. Reynolds)
                                    11

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           SUBLETHAL EFFECTS OF PETROLEUM ON MACOMA BALTHICA

                   M.S. Stekoll, L.E. Clement, and D.G. Shaw*
     The bivalve mollusc Macoma balthica has previously been proposed
as an indicator of oil pollution  (Shaw et a^l. , 1976).  This small clam
was selected for study because of its wide distribution and abundance
in Alaskan intertidal environments.  Macoma balthica is a surface deposit
feeder and suspension feeder, which would make it likely to concentrate
both suspended and stranded oil that might occur following a spill
(Brafield and Newell, 1961).  Such concentrated oil in the clams might
be further distributed throughout the ecosystem via migrating birds
that feed on M. balthica.

     Earlier experiments that subjected M. balthica to simulated oil
slicks showed mortalities at an applied rate of 5.0 vig/cm2 (Shaw e_t al.,
1976? Shaw et al., 1977).  Laboratory experiments indicated that ini-
tially buried clams would come to the surface when exposed to certain
levels of Prudhoe Bay crude oil in the water (Feder et a^L. , 1976) .
Macoma balthica, therefore, seems to respond to oil in its environment.
It was the purpose of the present investigation to show that M.
balthica is able to concentrate oil from an oil-sea water-dispersion
and to ascertain what effects the oil might have on membrane-bound
enzymes and other physical and chemical parameters of the clams.
METHODS

     Experimental clams were to be exposed to oil-sea water mixtures
for 3 weeks, followed by a 4-week recovery period.  For each treatment,
oiled and control, 900 clams were used.  Clams were divided into sam-
pling lots and placed into 2 cm x 10 cm glass Petri dish bottoms, each
containing 175g of silica sand and 15 clams.  The Petri dishes were
placed on an acrylic rack, which held 30 dishes in five levels.  Two
racks were used for each treatment.  The racks were set in exposure
chambers made of acrylic with dimensions of 24 cm x 66 cm x 30 cm.
*    Institute of Marine Science, University of Alaska, Fairbanks,
Alaska 99701.

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Each chamber was filled with about 33 to 35& of sea water or oiled sea
water and cooled by immersion in distilled water maintained at 10C.

     The clams used in this experiment were collected from the northern
end of Resurrection Bay near Seward, Alaska, in September 1976.  Clams
were maintained in Instant Ocean aquaria for 2 months prior to the
experiment.  The clams were allowed to depurate in clean sea water at
5C with aeration for 3 days just before exposure to the oil.  All
apparatus and sand that would come into contact with the clams were
either heated at 500C for 24 hr or extracted for several days in sea
water to remove unwanted hydrocarbons.

     The sea water used in this experiment was collected from Seward,
Alaska, and stored in 55 gal stainless steel drums at 4 until used.
The sea water in the exposure chambers was changed every other day.
About 35 of sea water were poured into 15 gal, all-glass aquaria and
aerated through gas dispersion tubes for 2 to 3 hours.  The control  (no
oil) was then siphoned into the previously drained exposure chamber.

     One liter of sea water was removed from the water to be oiled.  To
this, about 200 y& of Prudhoe Bay crude oil was added.  Addition of the
oil was through a bent tip attached to a Gilson Pipetman adjustable
pipet  (0 to 200 y).  The oil was dispensed in small increments directly
underneath a sonic probe  (Biosonik) set at 15% of maximum, causing
an oil-water dispersion.  Droplet size was determined to be in the range
of 1 to 5 ym.  Enough of this oil-water dispersion was re-added to the
sea water to give a final exposure concentration in the range of 1 to
2 yg of oil per milliliter of water.  This oil-water mixture was
siphoned into the exposure chamber.

     Water changes were performed every other day starting on day 0 with
the last change on day 18.  On day 21, each set of racks was trans-
ferred to a separate 25 gal Instant Ocean tank filled with clean sea
water and maintained at 8 to 10C with aeration.  The experiment was
terminated on day 49.

     Samples for hydrocarbon analysis were taken on days 0, 10, 21, 35,
and 49.  Ten dishes (150 clams) from each treatment were removed from
the tanks and screened to remove sand.  The clams were then depurated
in clean sea water for 24 hr before shucking for hydrocarbon analysis.
Sediment samples were collected on days 10 and 21 and frozen for later
analysis.  Water samples were collected at intervals (Exhibit 1)  and
extracted with hexane (see Appendix I for extraction and analysis pro-
cedures) .

     Samples for enzyme assays, glycogen, fresh and dry weights,  and
total lipid were collected on days 0, 7, 14, 21, 35, and 49.   Three
dishes were selected at random from each treatment chamber.  Clams were
screened from the sand and placed in beakers of clean sea water on ice.
For enzyme assays, 10 clams from each Petri dish were shucked and homo-
genized in 10.0 mi of 0.25 M sucrose, 1.0 mM Na2EDTA, pH 7.0 with a

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Virtis '45' homogenizer set at top speed for two, 10 sec bursts.   The
homogenate was filtered through gauze and further homogenized  in  a
Teflon-glass homogenizer at 1000 rpm for two strokes.  Part of the homog-
enate was frozen for later assay for glucose-6-phosphatase, Mg-ATPase,
and NA+-K+-ATPase.  The rest of the homogenate was centrifuged at 1000
x g for 10 min, and the supernatant was respun at 17,300 x g for  20 min
for a mitochondrial pellet fraction.  The pellet was resuspended  in
0.25 M sucrose, 1 mM EDTA, pH 7.0, and assayed for succinate dehydrog-
enase activity.   (See Appendix II for enzyme assays.)

          The remaining clams were allowed to depurate for 24 hr  in clean
sea water, then shucked into tared test tubes and frozen.  After  thawing
and weighing, these clams were lyophilized to dryness, reweighed, arid
assayed for either glycogen or total lipids.  (See Appendix III for
assays.)

        EXHIBIT 1:  HYDROCARBON CONCENTRATIONS IN UNEXPOSED
          CLAMS, CLAMS EXPOSED TO PRUDHOE BAY CRUDE OIL,
                  OILED, AND NON-OILED SEA WATER

               Clams (yg hydrocarbons/g wet weight)

             Fraction 1*                      Fraction 2*
Day	Oiled	Control	Oiled	Control
0
10
21
35
49

54.
74.
48.
38.

94 (4
52 (0
55 (7
96 (1
18.62
.8)
.6)
.0)
-7)

19.77
22.06
16.83
18.77

(3.
(5.
(1.
(3.

4)
0)
0)
6)

48.
54.
48.
46.

88 (4.
54 (5.
47 (9.
82 (14
14.08
8)
2)
0)*
.0)*
t-
21.20
37.30
35.16f
42.55

(5.0)
(3.4)

(4.1)*
                         Sea water  (yg/m)
        Time             Oiled                Non-oiled
          0 hr    0.64   (0.11) n =  4      0.014  (0.002) n = 2
   after  48 hr    0.086  (0.02) n =  6      0.020  (0.004) n = 2


 *    Values are means  (and standard deviations)  of three determinations,
 except  where noted, and quantified  from the chromatograms in the region
 bounded by C;^ and C%2 normal alkanes, using an  external standard.
 Fraction  one contains saturated alkanes, and fraction two contains aro-
 matic compounds, unsaturated hydrocarbons, and several other compounds
 of biological origin.

 t    n  =  1.

 f    n-  2.

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RESULTS AND DISCUSSION

     Part of our hypothesis  in this  experiment was  that the  oiled  clams
would show aberrations in  their membrane-bound enzyme  systems because
of the lipid-lipid associations between hydrocarbons and membranes
 (Roubal, 1974).  Since many  membrane-bound enzymes  need associated lip-
ids to function effectively  (White,  et_ al_.,  1973),  there was a possi-
bility that the oil  would  disrupt this association  (Manwell  and Baker,
1967; Roubal,  1974).  We also measured other parameters in the clams to
determine possible general effects on proteins,  lipids, glycogen,  and
fresh and dry  weights of the clams.

     The oiling treatment  in this experiment appeared  to have no dele-
terious effects on the clams.  The enzyme  assays showed no significant
differences between  the experimental group and the  controls  (Exhibit 2).
Succinate dehydrogenase levels remained at 15 to 25 nmoles/min/mg  pro-
tein.  The Na+-K+-ATPase pump varied from  7  to 12 nmoles Pjymin/mg pro-
tein with no differences between  the two groups.  The  Mg-ATPase levels
for both groups rose initially, peaking at day 21,  and then  fell back
again toward .day 0 levels.   Glucose-6-phosphatase remained fairly  con-
stant throughout the experiment.

     Measurement of  the other parameters also failed to show any signif-
icant differences between  the two groups of clams (Exhibit 3).  Both
groups showed  a slight decrease in dry/wet weight,  which was a result
of both a decrease in dry  weight  and an increase in wet weight.  The
protein/wet weight values  rose initially,  then dropped and leveled off
for each group.  There was a large variability in the  glycogen values,
making it difficult  to observe any overall  trend in glycogen content of
the clams.  The P^/protein values dropped  slightly  during the experiment
for both groups.  There was  also  no  difference between the control and
experimental groups  with respect  to  total  extractable  lipids.

     There were two  additional observations  that indicated some differ-
ence between the control and the  experimental groups.  The first was
the observation that the controls appeared to exhibit  more siphon  activ-
ity than the oiled clams.  We did not measure this  activity  quantita-
tively, but whereas  the control group extended their siphons in active
suspension feeding behavior  (Brafield and  Newell, 1961), the oiled
group displayed very little  siphoning behavior.   The second  observation
is that 19 control clams died  (gaping shells) and only 2 oiled clams
died during the  first 21 days.  The  reason for this relatively high mor-
tality in the  controls is  not known, but it may  be  related to oxygen
availability as a function of siphoning activity.

     When the  clams were extracted for hydrocarbons, however, there
were definite  differences  between the  two groups.   The oiled clams did
indeed take up and concentrate hydrocarbons.  Exhibits 4 and 5 indicate
the qualitative differences  between  the clams on  day 21.  Exhibit  1

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gives the values for hydrocarbons in the clams at different time peri-
ods.  The oiled clams retained hydrocarbons throughout the end of the
experiment.

     Levels of hydrocarbons in the control and oiled waters are shown
in Exhibit 1.  Sediment from both the control and the oiled treatments
contained very small amounts of hydrocarbons with no differences exist-
ing between the two.
                     EXHIBIT 2:  ENZYME ASSAYS
Day
0
0

7
7
14
14
21
21
35
35
49
49
Succinate
Oil Dehydrogenase
22.9
+ 21.4
17.0
+ 21.7
17.3
+ 15.6
24.0
+ 20.0
22.8
+ 26.7
19-9
+ 23.2
 2.
 3.
 2.
 2.
 0.
 1.
 3.
 2.
 2.
 3.
 1.
 2.
1
7
7
3
9
7
8
7
4
8
2
4
Glucose-6-
Phosphatase
2.13
2.47
1.82
1.65
1.78
2.11
2.19
1.98
1.82
1.73
1.94
1.54
 0.
 0.
 0.
 0.
 0.
 0.
 0.
 0.
 0.
 0.
 0.
 0.
25
24
32
46
38
17
55
34
19
06
13
32
MgATPase
32.
30.
41.
41.
39.
39.
52.
47.
44.
47.
38.
39.
9 
6 
2 
6 
6 
7 
3 
1 +
5 
7 
9 
4 
2.9
5.7
2.9
6.9
2.6
4.7
6.9
4.5
2.4
5.8
3.2
2.7
Na+-K+-ATPase
8.9
10.7
8.0
7.1
10.1
9.0
8.7
9.0
9.2
8.9
8.1
12.6
 0.
 3.
 1.
 4.
 1.
 0.
 1.
 1.
 0.
 2.
 3.
 0.
6
5
7
1
4
2
8
9
4
7
0
9
Note:  Assays were performed in triplicate on three sets of ten clams
each  (N=3).  Rates are expressed as nanomoles/min/mg protein  S.D.

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                   EXHIBIT 3:   SOME PARAMETERS  MEASURED ON  MACOMA  BALTHICA
DURING 49-DAY OILING EXPERIMENT
Day Oil
0
0 +
7
7 +
14
14 +
21f
21 +
35
35 +
49
49 +
i*
Pi1
Protein
(nmoles)
mg
235  28
256  28
212  4
222  14
212  8
208  10
221  18
210  4
219  16
221  7
193  6
193  10
*
Protein
Wet Weight
g
67.5  8.0
68.9  5.8
82.6  1.8
76.8  7.1
78.7  1.7
75.5  3.4
64.5  2.9
66.0  3.8
69.1  0.7
70.8  2.9
68.7  1.8
67.6  1.4
Dry
Wet
"(n)
(15)
(15)
(14)
(15)
(14)
(15)
(12)
(13)
(9)
(14)
(14)
(14)
Weight
Weight
20.6
20.7
20.2
20.8
21.0
20.4
20.2
20.5
18.9
18.8
15.3
15.3
 1.1
 0.6
 0.9
 1.0
 1.7
 0.8
 1.1
 1.2
 1.3
 1.2
 1.1
 0.7
Glycogen
Wet
(n)
(8)
(8)
(7)
(8)
(7)
(8)
(6)
(6)
(5)
(7)
(7)
(7)
Weight
incr
CT
10.0
7.0
5.2
8.4
12.5
9.5
7.1
6.8
6.3
7.4
3.0
3.9
 4.6
 3.9
 4.2
+ 2.3
 2.1
 5.5
 4.6
 3.5
 5.0
 4.6
 1.4
 2.9
Lipid
Wet
(n)
(6)
(6)
(7)
(7)
(7)
(7)
(6)
(7)
(4)
(7)
(7)
(7)
Weight
g
16.7 
18.5 
18.1 
14.5 
17.4 
16.6 
17.2 
19.8 
18.9 
17.8 
14.8 
15.2 

3.0
2.3
4.0
2.6
4.0
3.3
4.1
4.2
4.9
2.9
1.7
3.4
From three sets of 10 clams each  S.D.
Oiling regime stopped at

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EXHIBIT 4:  GAS  CHROMATOGRAMS OF ALKANE FRACTIONS FROM DAY 21  CLAMS
EXHIBIT 5:  GAS CHROMATOGRAMS
  OF AROMATIC FRACTIONS  FROM
  DAY 21 CLAMS
                                                          oiled
                                                           control

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     Although the clams concentrated portions of Prudhoe Bay crude oil
in this experiment, we could find no measurable differences between the
oiled and the control clams with respect to enzyme activities or other
measured parameters.  The relatively high mortality of the control
clams, however, indicates that the experimental set-up induced a stress
on the clams apart from the oil.  This stress may have been the lack of
aeration between water changes and/or the absence of food for the clams.
The oiled clams did not show this mortality perhaps because of their
lower filtration activity, which may be an indication of lowered meta-
bolic functioning.  A decrease in metabolic activity would certainly
place a smaller demand on the environment for survival.

     It is our feeling that longer exposure periods to oil are needed
in order to see any sublethal effects of oil on the clams.  A flow-
through system would reduce the stress and mortalities caused by the
water change procedure.  Such a system is currently being developed
in this laboratory.
REFERENCES
Brafield, A.E. and G.E. Newell.  1961.  "The Behavior of Macoma
  balthica  (L.)," J. Mar. Biol. Assoc. U.K.  41:81.

Feder, H.M., L.M. Cheek, P. Flanagan, S.C. Jewett, M.H. Johnston,
  A.S. Naidu, S.A. Norrel, A.J. Paul, A. Scarborough, and D.G. Shaw.
  1976.   "The Sediment Environment of Port Valdez, Alaska and the
  Effect  of Oil on this Ecosystem," final report on Project R800944
  to the  Environmental Protection Agency.

Manwell,  C.M. and C.M.A. Baker.  1967.  "A Study of Detergent Pollution
  by Molecular Methods:  Starch Gel Electrophoresis of a Variety of
  Enzymes and Other Proteins," J. Mar. Biol. Assoc. U.K.  47:659.

Roubal, W.T.  1974.  "Spin-labeling of living tissue;'a method for
  investigating pollutant-host interaction."  ^n F.J. Vernberg and
  W.E. Vernberg, eds., Pollution and Physiology of Marine Organisms,
  Academic  Press, New York:   367.

Shaw, D.G., A.J. Paul, L.M. Cheek, and H.M. Feder.  1976.  "Macoma
  balthica:  An Indicator of  Oil Pollution," Mar. Poll. Bull. 7:29.

Shaw, D.G., L.M. Cheek, and A.J. Paul.  1977.  Estuarine and Coastal
  Marine  Science.  In press.

White, A.,  P. Handler, and E.L. Smith.  1973.  Principles of Biochem-
  istry,  McGraw-Hill, Inc., New York.

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APPENDIX I:  ASSAYS FOR HYDROCARBONS IN
CLAMS, WATER AND SEDIMENT	

     General

     All solvents used were redistilled in glass and then checked for
purity by gas chromatography.

     All glassware was washed with soapy water, rinsed in distilled
water, and rinsed with organic solvents prior to use.

     Samples were concentrated on a rotary evaporator to approximately
2 mi.  They were then concentrated to final volume under a stream of
dry nitrogen.

     Gas chromatography  (GC) was performed using a Hewlett Packard
model 5710A gas chromatograph with flame ionization detectors.  GC out-
put was recorded on both an HP model 3380A integrator and a Sargent
model SR recorder with a disc integrator.  Sample components were sepa-
rated using a 12 ft x 1/8 in. column packed with 3% OV101 on 100/120
chromosorb W. Oven temperature was programmed from 80C to 280C at
8C/min.  Carrier gas was nitrogen at 50 m/min.  Individual peaks were
quantified using the electronic integrator, the total chromatogram by
disc integration.  Both integrators were calibrated using an n-alkane
standard.

     Clams

     Each group o.f approximately 50 clams was thawed and then removed
from their shells to a preweighed 50 m centrifuge tube.  Wet weight
was determined, 5 mi of 8N NaOH was added, and the tube sealed with a
teflon-lined cap.  The tubes were then placed in a 90C water bath for
3 hr, and were shaken every hour.  After digestion, 15 m of benzene
were added, the tubes shaken vigorously and centrifuged at 2000 rpm for
10 min.  The benzene phase was pipetted off, and the extraction pro-
cedure repeated twice using 10 mi benzene and 5 m water, saturated
with NaCl.  The three extracts were combined and dried over Na2SOi4.
The benzene extract was concentrated to approximately 0.5 m and placed
on a 0.9 cm x 22 cm column packed with 5 g of 6% deactivated alumina
over 5 g of 5% deactivated silica gel.  The column was eluted with 20
m of hexane  (fraction 1) and then with 20 m of benzene (fraction 2).
Fraction 1 was concentrated to approximately 0.2 mi, fraction 2 to
approximately 0.5 m, and a 5 m aliquot of each was gas chromato-
graphed .

     Water

     Hydrocarbon content of freshly oiled sea water was determined on
days 1, 5, 7,- and 9.  The amount remaining in this sea water after the
48 hr exposure period was determined on days 3, 5, 7, 9, 11, and 19.
The hydrocarbon content of unoiled sea water was determined before

-------
exposure on days 3 and 17, and after 48 hr/exposure on days 3 and 19.
Sample sizes were 500 m of freshly oiled sea water, 1000 m& for oiled
sea water after 48 hr, and 1000 ml for all unoiled sea water.  The
sample was extracted three times with a 5% sample volume of hexane.  The
hexane extracts were combined and dried over ^280^.  The hexane extract
was concentrated to approximately 0.5 mi, and a 5 m aliquot gas chroma-
tographed.

     Sediment

     A lOOg sample of sand was collected after screening from the oiled
chamber and from the control chamber on days 10 and 21.  Each sample
was extracted with 250 mil of refluxing methanol with stirring for 24 hr.
The hydrocarbons were then partitioned into hexane; the hexane extract
was dried over Na2SOt| and concentrated to approximately 1 m.  A 5 m
aliquot was gas chromatographed.
APPENDIX  II:  PROCEDURES FOR ENZYME ASSAYS

     Glucose-6-Phosphatase  (Baginski et_ a_^., 1974; Hiibscher and West,
1965)
     Assay Mix

	Treatments    A    B	C    D

1.   Sucrose/EDTA/KF*       0.10 ml                 +    +    +    +

2.   Cacodylate buffer      0.10 ml                 +    +'   +    +

.3.   Test solution          0.10 ml                 (a)   (b)   (c)   (d)

     keep on ice -  then at  t=0 add

4.   G-6-P                  0.10 ml                 +    +    +    +
Mix  (vortex)  and  incubate  30 min  at  30C.  Stop each reaction with 2.0
m of cold TCA/ascorbate  (10%,  2%)  (W/V) .  Put on ice for 2-3 min, then
centrifuge for  3  min  to precipitate  protein.  Remove 1.0 m of the
supernatant to  assay  for phosphate.
 *   To  make  solutions:

 1.   Sucrose/EDTA/KF

        16  mM Na^EDTA

         8  mM KF

       250  mM sucrose  -  adjust  pH  to 7.0 with HCl.

 2.   Cacodylate buffer (0.10  M)

       0.10 M cacodylic  acid, pH 6.5      '
                                10

-------
      dissolve 0.138 g cacodylic acid in H20.  Adjust pH  to  6.5 with

      NaOH.  Dilute to 10 mi.  Keeps 1 week.

 3.    G-6-P (0.10 M)

      Dissolve 0.282 g Na G-6-P in 10 m H2O.  Store cold.   Keeps  1 week.

 4.    Test solutions:

      (a) Blank:   0.100 itiA of 0.25 M sucrose/1 mM/EDTA

      (b) PI STD:  0.020 m& of 10 ymole/m K^HPOi^ + 0.080 m of sucrose/
          EDTA (0.25 M/lmM).

      (c) Control:  0.100 m of crude homogenate added after TCA/ascor-
          bate
      (d) Crude homogenate:  0.10 m of the crude homogenate.

 KF and EDTA are included to inhibit alkaline and acid phosphatous
 (Hiibscher, 1965) .

 Na+-K+-ATPase Assay (Jorgensen, 1974; Skou, 1957;  Dunson and Dunson,
 1975)
                                        Treatments (in triplicate)
Add
1.
2.
3.
4.
5.
6.
7.
8.
9.
in order
H20
0.5 M imidazole HC1
pH 7.5
0.1 M MgCl2
10 mM Na2EDTA
1 M NaCl
1 M KC1
10 mM ouabain
Test solution
75 mM ATP
Total Volume
mS,
0.430
0.100
0.040
0.050
0.020
0.020
0.200
0.100
0.040
1.000
A B C D E
+ + + + +
+
+ + + + +
+
+ + + +
H-
-
(a) (b) (c) (d) (e)
+ + + + +
note: (-)=H2O in place of
                                              solution.
Tubes are kept on ice until ATP is added, at which time tubes are vor-
texed and set in 20C water bath  for  10  min.
Test solutions

(a)  Blank 0.100 mH of sucrose/EDTA (0.25 ImM)

                                 11

-------
(b)   Pi STD 0.020 m of 10 ymoles/m phosphate
            0.080 mH of sucrose/EDTA  (0.25 M/lmM)

(c)   Control - 0.100 mJl of crude homogenate  (diluted with sucrose/
     EDTA if necessary) added after reaction is stopped with TCA/
     ascorbate.

(d)   Total ATPase - 0.100 m of crude homogenate

(e)   Mg-ATPase - 0.100 m of crude homogenate

Stop reaction with 2.0 m of cold ascorbate/TCA (2.5%/12.5%).    (W/V) .
Vortex.  Set on ice 5-10 min.  Centrifuge precipitated protein 3-5 min
on clinic-1 centrifuge (^2400 rpm).  Remove 1.0 m of supernatant for
phosphate analysis.  Na+-K+-ATPase is difference between total ATPase
and MgATPase.

     Succinate Dehydrogenase  (Singer, 1974; Bonner, 1955)

     Preparation of Reagents:

1.   KCN 0.1 M; to 70 m of cold H20 add 0.6512 gm KCN. Then add 8.5 m
     of cold 1 N HCL to neutralize KCN,  Keep stoppered for 10 min to
     readsorb KCN fumes.  Then make up to the mark with cold ^O.  Store
     cold.  Reagent is good for 2 days.

2.   K3FE(CN)6 0.01 M; to 100 m H2O add 0.3293 gm of potassium Ferri-
       cynide  (Sigma 99%).  Keep in a dark bottle protected from light, to

3.   Na succinate 0.20 M; adjust to pH 7.5 at room temperature.

4.   KPO4 buffer, 0.30 M, pH 7.5 at room temperature.

     Assay Procedure;

Into 3.0 m cuvettes add in order:

                                           Final Cone.
!-   KCN 0.1 M               0.300 m&      0.01 M
2.   K3Fe(CN)6 0.01 M        0.400 mi      0.33 K10~3 M
3.   Na succinate 0.2 M      0.200 m      1.33 x 10~2 M

4.   KP04 buffer pH 7.5      1.000 m&      0.100 M

5.   H2O                     0.850 mH

6.   Diluted mitochondrial
     prep.                   0.250 mj,
                             3.000 mJl
                                 12

-------
Add mitochondrial prep after all reagents have  come  to room temperature
(20C).  Place cuvette in spectrophotometer  at  20C  and measure AOD at
420 nm.

    Measure temperature at end of the run.

    A blank (control) is run substituting water  for Na-succinate.
     This is subtracted from the  (+) succinate  cuvette.

    AOD should decrease at about 1-10 x 10"^ OD/min.

Note:  One mole of succinate reduces 2 moles of Fe(Cn),-.
       e  for Fe(CN),. = 1.0 at 420 nm.
        mM           6
REFERENCES

Baginski, E.S., P.P. Foa, and B. Zak.  1974.   "Glucose-6-phosphatase,"
  In H.U. Bergmeyer, ed., Methods in Enzymatic Analysis, Academic Press,
  New Yor, 11:876.

Bonner, W.D.  1955.  "Succinic Dehydrogenase."  In^ S.P- Colowick and
  N.O. Kaplan, eds., Methods in Enzymology, Academic Press, New York,
  11:722.

Dunson, M.K. and W.A. Dunson.  1975.  J. Comp. Physiol.  101:89.

Hlibscher, G. and G.R. West.  1965.  "Specific Assays of Some Phospha-
  tases in Subcellular Fractions of Small Intestinal Mucosa," Nature,
  205:799.

Jorgensen, P.  1974.  "Isolation of  (Na +K )-ATPase."  ^n_S. Fleisher
  and L. Parker, eds., Methods in Enzymology, Academic Press,  New York,
  XXXII:277.

Singer, T.P.  1974.  "Determination of the Activity of Succinate, NADH,
  Choline, and ot-Glycerophosphate Dehydrogenases," In D. Click, ed.,
  Methods of Biochemical Analysis, J. Wiley and Sons, New York,
  XXII:123.

Skou, J.C.  1957.   "The  Influence of Cations on an Adenosine Triphos-
  phatose from Peripheral Nerves," Biochem. Biophys. Acta, 23:394.
                                13

-------
APPENDIX III;  ASSAYS FOR PHOSPHATE, GLYCOGEN, AND TOTAL LIPIDS

     Phosphate Assay (Baginski et_ al., 1974).

     To 1.0 m of incubation supernatant, which contains VL.67%
ascorbate and 8-9% TCA add:

     0.50 m 1%  (W/V) ammonium molybdate in H O.  (Note:  HO must be
     very pure.)

     Mix contents immediately  (vortex).  Wait exactly 5.0 min  (or some
     other exact time between 3-10 min).  Then add:

     1.00 m of sodium arsenite/sodium citrate  (each 2% W/V) in 2%
     acetic acid  (V/V).

     Mix contents immediately and let stand 15 min or longer.  Read
     absorbance at 700 nm vs blank containing 1.0 m HO + Pi reagents
     (or 1.0 m of diluted ascorbate/TCA).

     Color is stable for several hr.

     Assay for Glycogen (Seifter,  1950)

     Select 5-10 clams.  Set in clean sea water for 24 hr at 4C to
attempt to eliminate sand from the clams.  Add 1.0 m of 30% KOH to
dried clams in test tubes.  Digest clams in a 95C bath with marble
caps.  Incubate 40-60 min until all  of clam is digested.

     Cool digest and add 1.25 m of  95% EtOH.  Vortex to mix.  Heat to
gentle boil on water bath  (^95C), using marbles to cap the tubes.
Cool.  Centrifuge at 2300 rpm for 15 min (International model K).  Pour
off supernatant.  Wash pellet by resuspending in 1.0 m HO + 1.25 m
EtOH.  Heat, cool, and centrifuge again.  Pour off supernatant.  Allow
tubes to drain inverted on paper towels.

     Resuspend pellet in 5.0 m of H2O.  Heat to dissolve glycogen.
Cool to room temperature.  Remove 0.5 m to assay for carbohydrate by
the anthrone method.  May store frozen, if necessary.


     Express results as mg glycogen/gm fresh wet.
                                  100 x A620 f sample
        mg glycogen in aliquote =
                                  1.11 x A620 of 1 m9 std
                                 14

-------
     Anthrone Assay (Spiro, 1966)

     Reagents:

     280 rnH H2O
    +720 mJl cone I^SO. (reagent grade) .
     While still warm, add:

     500 mg anthrone
      10.0 g thiourea

Mix to dissolve.  Cool and store at 4C.  Age 7 hr.  Reagent will keep
about 2 weeks.

     Procedure:

     Use any test tube >10 m capacity
     range 20-200 yg hexose in 1.0 mX, HO
     1.0 m sample on ice
     5.0 m cold anthrone reagent

     Vortex to mix.  Keep on ice until all tubes are ready.  Cap with
marbles.  Place in boiling 1^0 for exactly 10 min.  Remove and cool in
H20 bath to room temperature.  Let stand 20 min.  Read absorbances at
620 nm.

     Lipid Extraction (Radin, 1969)

     Add 100 rn of distilled H2O to lyophilized clam in 14 x 100 mm test
tube.  Allow to rehydrate for 20 min.  Grind up clam with clean glass
rod.  Rinse rod 3 times with 0.5 m benzene-MeoH (2:1 v/v).  Add rinses
to tubes containing clams.  Vortex.  Sonicate with small needle probe
for 5 seconds at 'min1 and 5 sees at 50%.  Cap tubes with marbles and
incubate for 20 min at 50-55C.  Centrifuge and place supernatant into
clean large test tubes.  Wash residue two times with 2.0 m and then
1.0 m of benzene:MeoH, 2.1  (v/v).  Add washings to supernatant.

     Add 1.0 m& of 0.75% NaCl to extract water soluble lipids.  Vortex.
Centrifuge.  Remove upper layer; place in tared vial (if there is an
emulsion or more than 2 layers, add more NaCl).  Re-extract water layer
two times with 1.0 m benzene.  Combine all benzene fractions in the
tared vial.

     Evaporate contents of vial under N2 stream.  When dry, lyophilize
to remove water residue.  Re-weigh dried vial to obtain weight of total
lipids in clam.
                                15

-------
REFERENCES

Baginski, E.S., P.P. Foa, and B. Zak.  1974.  In_H.U- Bergmeyer, ed.,
  Methods of Enzyme Analysis^ Academic Press, New York, II.; 876.

Radin, N.S.  1969.  "Preparation of  Lipid Extracts."  In_ J..M.
  Lowenstein, ed., Methods  in Enzymology, Academic Press, New York,
  XIV:245-254.

Seifter, S. , S. Dayton, B.  Novic,  and E. Muntwyler.   1950.   "The Estima-
  tion of Glycogen with the Anthrone Reagent," Arch Biochem.  (Biophys.),
  25_:191.

Spiro, R.B.  1966.  In E.F. Neufeld  and V.  Ginsburg,  eds. , MethodsjLn_
  Enzymology, Academic Press, New  York, VIII;3.
 DISCUSSION

      Dr.  McErlan:   If a  more  natural  system was  established  that  allowed
 the  sediment  to  be  contaminated around the  animal,  an entirely different
 result might  be  obtained.   Generally,  the Macoma uses two  siphons, which
 pick something off  the interface of the water  column at  the  bottom.
 This method is its  primary means of obtaining  food.  It  is much quicker
 than straining water.

      Dr.  Stekoll;   The Macoma feeds both ways.  They stick their  siphons
 out  as if it  is  in  suspension and they deposit feed.  However,  we are
 mainly concerned with the effects of  the oil   not with their  feeding
 mechanisms.  Our experiment did not demonstrate any difference between
 the  oiled and controlled clams.

      Dr.  Dieter: Did the oiled sea water contain  about  0.5  mg/gm of
 alkanes,  and, did your clams, subtracting the  values from the controls,
 have from 20  to  50  mg/gm of alkanes?

      Mr.  Stekoll:   They  concentrate approximately  80 times.

      Dr.  Dieter:  In analyzing petroleum hydrocarbons  in the clams,  what
 is your  percent  recovery?

      Mr.  Stekoll:   70 to 90.   We put in an internal standard of  C-14,
 C-21, C-30, to get  our recovery back  and correct for  that.

      Dr.  Rogerson:   Your recoveries from the  aromatic  fractions  appear
 to be surprisingly  low.   Do you use fully activated silica and aluminum?

      Mr.  Stekoll:   Both of them were partially deactivated.   We added
 approximately 6% for alumina, and 5% for the  silica gel.
                                 16

-------
     Dr.  Rogerson;  We have noticed that the heavier aromatics when more
fully activated don't move.

     Mr.  Stekoll:  True.  We are in the process of redoing our whole
extraction procedure for recovery.  We have also been having problems
with the methylester formation.  We eliminated the methanol in the
siphonification, and then had problems with emulsions.  So we had to
extract with benzene instead of hexane, using a centrifuge.
                                17

-------
               THE BENTHIC COMMUNITY OF A NATURAL OIL SEEP
                       IN THE SANTA BARBARA CHANNEL

               Robert B. Spies, Paul H. Davis, and Thomas M. Distler*
     Little information is available on the effects of petroleum hydro-
carbons on soft bottom benthic communities.  Post-spill surveys, such as
after the 1969 Santa Barbara Oil Spill  (Straughan, 1971) are generally
made against a background of insufficient data and, therefore, are only
able to detect the most gross effects.  A notable exception has been the
follow-up work on the West Falmouth Oil Spill  (Michael et al., 1975).
Often, other complicating factors, such as the extensive use of deter-
gents  (Smith, 1970), have made it difficult to separate the effects of
petroleum hydrocarbons.  Since spills occur unpredictably and are ephem-
eral in behavior, the systematic study of their effects is extremely
difficult.  There is usually insufficient background data, and the lo-
gistical problems are difficult.  Sources of chronic hydrocarbon pollu-
tion should provide a more stable setting for  studying such effects.  In
addition, if the best estimates of input rates of petroleum hydrocarbons
to the marine environment  (National Academy of Science, 1975, p. 6) are
examined, it can be seen that chronic sources exceed accidental sources.
Therefore, we believe that a thorough study of chronic hydrocarbon pol-
lution of soft bottom communities is needed.

     The profuse offshore oil seeps of the Santa Barbara Channel pro-
vide an excellent natural laboratory for such a study.  Previous work
has established the location, nature, and estimates of flow rates of
these seeps  (Rosen et al., 1959; Wilkinson, 1971; Vernon and Slater,
1963; Mikolaj et al., 1972; Allen and Schlueter, 1969; Allen et a^L.,
1970).

     Detailed information provided by Allen et al. (1970)  on a small
group of profuse seeps near Goleta,  California, led us to investigate
these as a possible area for our study.   After some exploratory dives,
we found that one relatively isolated seep, 800 to 900 m off Isla Vista,
is located in an area of a homogeneous-appearing sand bottom.  The sedi-
ments are extensively contaminated with crude oil and, at 15 to 16 m
depths, it is shallow enough to employ diving for intensive sampling and
observations.
*    Lawrence Livermore Laboratory, University of California, Livermore,
Cali fornia  94 5 5 0.
                                 19

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     The only ecological work in the open literature concerning the seep
is a report of the absence of ovicell hyperplasia in bryzoans from the
vicinity of Coal Oil Point  (Straughan and Lawrence, 1975).  We report
here the preliminary results of a study of benthic community composition
in this oil seep and a comparable nonseep area nearby.  These are the
first results of a study of temporal variation in the Nothria-Tellina
community, which is a large-scale feature of the northern Santa Barbara
Shelf  (Jones, 1969).  It is intended that data from the comparison sta-
tion serve as a measure of natural temporal variation in this community.
METHODS

     To find a suitable study site, we made a series of dives in the
group of seeps running southwesterly from Coal Oil Point, Goleta, Cali-
fornia.  The sand overburden in this area was generally thin and vari-
able in water depths to 21 m.  Shale outcroppings were common, and it
was apparent that extensive sampling would be required to adequately
characterize the benthic community.  We finally settled on the nearby
Isla Vista Seep  (Cd in Mikolaj, 1972) because of the larger sand over-
burden in shallow water and, consequently, more homogeneous bottom.  A
nonseep comparison station was selected on a similar bottom type approx-
imately 1,408 m  (7/8 mile) to the east  (Exhibit 1).  We are well aware
that this second station is not an absolute control.  We have seen small
amounts of weathered tar in the sediments there, and occasionally a sur-
face slick has coated our buoy.  However, we have never seen fresh oil
in the cores from the nonseep station, and chemical analyses bear out
the large differences in hydrocarbons at the two areas.  We did not want
to establish the nonseep station too much farther away because of the
possibility of other physical and chemical factors influencing community
composition.  As it is, both stations are 15 m deep, have identical
fine sand sediments, and are equidistant from a prominent headland.  Our
observations are that currents, surge, suspended particulates, and light
are also always similar in the two areas.

     Each station is marked with a buoy for accurate resampling.  We
have had trouble losing buoys in storms.  Consequently, the original
seep station of December 1975 was reestablished in February and again in
April 1976, using identical compass fixes on local landmarks.  We have
been successful in relocating the nonseep station with underwater search
techniques and, consequently, it has been continuously sampled.  During
later reconnaissance, we have relocated the second seep station relative
to the present station.  It is approximately 53 m west and at the same
depth.

     We now have a sturdy, all-stainless steel buoy system consisting of
a fence post anchor, 3/16 in. cable, and a large foam-filled buoy.  The
fence post anchor has a 4 in. disc and is screwed at least 65 cm into
the bottom.  Spatial continuity in seep station samples begins April
1976.
                                20

-------
EXHIBIT 1:  MAP OF STUDY AREA
     Samples are taken within a 50 m'2 area at each station.  The area
has been subdivided into two hundred 0.25 m2 quadrates.  Every 2 months,
10 quadrates are sampled at each station.  The quadrates are selected
using a random number table.  This system allows us to record the loca-
tion of every core taken during the study to eventually determine if
there are any small-scale spatial patterns in the local community.  Our
present plans are to sample each station every 2 months for at least 18
months and then to continue sampling on a less rigorous schedule for a
number of years.
                                           2
     The samples are vertical cores 0.018 m  in area and 20 cm deep.
The coring device is a 3 Ib coffee can, open at the bottom and punched
with a hole in the top to accept a No. 9 rubber stopper.  The can is
pushed all the way into the sediment, the stopper inserted, and then
the whole can is extracted.  The open end is capped with a plastic lid.
The samples are sea water sieved on a 0.5 mm mesh screen within 4 hours
of collection.  The sieve contents are then preserved in 10% neutralized
formalin and sea water.  In the laboratory, samples are washed and elu-
triated with fresh water, which concentrates most worms and crustaceans
in a small volume separate from the sediment.  The eluted and sediment
fractions are then sorted in large trays under low-power stereomicro-
scopes.  Polychaetes, crustaceans, molluscs, and miscellaneous groups
are separated and preserved in a 70% ethyl alcohol-5% glycerine solu-
tion.

     Biomass determinations were initiated with the February 1976 sam-
ples.  Determinations are made on each core for each group of rough-
sorted organisms.  An attempt is made to separate particularly large
organisms and weigh them separately.  Organisms are removed from the
alcohol preservative, blotted dry, air dried for 10 min, and weighed.

     All organisms, with a few exceptions, are identified by species and
counted for each core.
                                 21

-------
     Community diversity H' is estimated  according  to  Shannon-Weaver
 (1963) using the approximation:

           s n.      n
     H' ' - IT Iog2 IT

 in conjunction with Pielou's measure of evenness J  (1966a) :
                       _
          H max      Iog2s  '


where

     N = total number of individuals  (all species)

     n- = number of individuals of each species

     s = number of species.

     Smaller, 5 cm diameter cores were used to obtain samples for  sedi-
ment size-class distribution and hydrocarbon analysis.  These are  found
in about the top  22 to  25 cm of sediment.

     For hydrocarbon analysis of sediments, each core is thoroughly
mixed and subsampled.   The  subsample  is acid-treated, water-washed, and
freeze-dried followed by extraction with toluene-methanol for 300  cycles
in a soxhlet apparatus  (Rohrback and  Reed, 1975).  Total extractable
material is determined  gravimetrically on a microbalance.

     Sediment-size-class analyses were performed using sieving and
modified Emory settling-tube methods  following the method of Buchanan
and Kain  (1971) .


RESULTS

     We describe  the results of our analysis below.

     Hydrocarbon  Measurements

     The results  from April and June  1976 are given in Exhibit 2.  These
were collected from the current location of the seep station and are not
taken from the same exact location as the faunal data reported here;
however, they are representative of the differences in hydrocarbon com-
position of sediments in the two areas.  It can be seen that there is a
threefold to fivefold difference between the two areas.  Excluding one
sample with a large tar lamp, the mean values for the two sampling peri-
ods from the seep station are very close, and the deviations are only
about 20%.  The preliminary data also suggest that the sediment
                                 22

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hydrocarbons  at the nonseep station are more variable with time, perhaps
as a result of irregular local  fallout of weathered tar.
       EXHIBIT 2:  SEDIMENT EXTRACTABLES (% of dry  sediment)
            Seep
                                       Nonseep
April
0
0
0
1
1976
.70
.81
.51
.83
 0
 0
 0
- 2
03
03
03
0
~x = 0.67  0.15 0
0
77* 0
15 
15 
14 
10 
0
0
0
0
01
01
01
01
                                      x = 0.14  0.02
        June 1976

           0.79  0.02

           0.49  0.02

           0.81  0.02

           0.56  0.02
         * = 0.66  0.16
0.28  0.02

0.17  0.02

0.18  0.02

0.22  0.02

0.21  0.05
         *Tar lump In sample.

     Sediment-Size Analysis

     The weight  loss after drying 24 hr  from two small sediment-core
samples  from each station varied by only 2%.   Therefore, the water  con-
tent is  assumed  to be essentially the  same  at each station  (25%).

     The results of the sediraent-grain-size analysis are given  in
Exhibit  3  for these four samples.  As  can be seen, the sediments are
very similar.  There is about a 5% to  7% difference in the  amount of
very fine  sand  (125 to 62y) , probably  because of the slight skewing by
small bits of weathered tar in the upper size ranges of the seep sedi-
ments.   This analysis confirms our earlier subjective judgments of  the
similarity of the two stations.
                                  23

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EXHIBIT  3:   SEDIMENT  SIZE
  DISTRIBUTION AT SEEP  (A)  AND
  NONSEEP (B) STATIONS
                                      fi 30 -
                                        10 -
60


50


40


30


20


10
                                          . B
                                             1000  250  62 62/   39/
                                              rr' 15-6   
                                                  Sand    ,	._,
                                                         Silt clay
                                                Sediment size (y)
                                             1000  250
          Sand
              62 62/  39/
              -" 15.6   0
                 Silt clay
       Sediment size (y)
      Biological  Measurements

      The December and February  samples have  been completely processed.
The  number of  species, individuals, and diversity for both stations  and
sampling periods are given in Exhibit 4.
                                   24

-------
          EXHIBIT 4:   COMMUNITY PARAMETERS SEEP (NONSEEP)
                          Dec 1975       Feb 1976       Total
No. of species
No. of individuals
Total individuals
Diversity (H1)
Evenness (J)
153
2900
(126)
(1302)
4202
3.70
0.74
(3.72)
(0.77)
106
1823
(103)
(1264)
192
4723 (2565)
3087
3.59
0.77
(3.52)
(0.76)


     A number of trends are immediately evident:

1.   The number of individuals is double at the seep station.

2.   The number of species is about 15% higher at the seep station for
     both sampling periods.

3.   There is a sharp reduction in both species and individuals at both
     stations in February 1976.  In addition, the mean biomass at the
     seep station, 1.53  0.85 g, is higher than at the nonseep station,
     1.02  0.47 g (February 1976), but these differences are not sig-
     nificant 


-------
EXHIBIT 5:  DISTRIBUTION OF
  INDIVIDUALS AND  SPECIES IN
  COLLECTIONS FROM BOTH STATIONS
   [ (   ) = Cumulative % of
  total species]
                                            165

                                            135

                                            105

                                             75

                                             45

                                             15
                                                nii	r
                                                       Seep station
                                                         (90%)
                                                             n	r
  250
                                                    1250   2250   3250
                                                    Number of individuals
                                                       Nonseep station
                                          <" 135 -
15
                                                                     4250
           'Core numbers,
           taken at random
       625    1125   1625   2125
        Number of individuals
the  seep  station,  and one species has the same.   In February,  there were
22 such species with 17 (77%)  having greater densities  at the  seep sta-
tion and  one species having the same.

     To compare the two stations further, a Spearman  rank correlation
analysis  was performed on the 26 co-occurring common  species from Decem-
ber.  The results indicated a significant  (p =  0,01)  positive correla-
tion (rs  = 0.64).   A similar result was observed for  the February data,
thus supporting the hypothesis that the community structures of the two
areas  are basically the same.

     A list of the 10 most common species from  each station in the
December  samples is given in Exhibit 6.  Included are the mean densi-
ties,  standard errors,  and an estimate of the number  of samples required
to achieve a 20% degree of precision (Elliott,  1971).   This last measure
is an  indication of which species will be amenable to more detailed pop-
ulation studies under the current sampling scheme.

     Of the 10 most common species listed for the seep station in
Exhibit 6,  the following species showed a 300%  or more increase over the
control station:  Mediomastus californiensis, Tellina modesta, Oligo-
chaetes,  and PraxilTella affinis pacifica.These  are  all deposit
feeders.
                                 26

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 EXHIBIT  6:   TEN  MOST ABUNDANT SPECIES  (TAXA) FROM DECEMBER 1975

A.
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
Species
Collected at seep station
Mediomastus californiensis
Tellina modesta
Oligochaetes
Nematodes
Chaetozone neosetosa
Prionospio pygmaeus
Praxillella affinis pacifica
Euphilomedes sp .
Nepthys caecoides
Lumbrinereis tetraura
Density/Core
29.
29.
30.
22.
18.
10.
9.
9.
8.
6.
8 
2 
7 
3 
1 
0 
8 +
4 
1 
6 
9
13
20
14
10
7
6
4
5
3
.22
.5
.2
.3
.9
.9
.6
.1
.9
.9
2
s
" D2x2
2
5
10
10
8
15
11
4
13
8
*
, (D = 0.20)
.4
.3
.8
_ 2
.5
.7
.2
.8
.3
.7
        B.  Collected at nonseep station

         1. "Prionospio pygmaeus           16.7  6.1            3.3

         2. Nematodes                   14.3  9.3           10.4

         3. Chaetozone neosetosa            8.6  4.0            5.3

         4. Mediomastus acutus             7.6  3.5            5.2

         5. Mediomastus californiensis       7.4  3.3            5.0

         6. Tellina modesta               6.1  5.0           16.9

         7. Euphilomedes sp.              4.9  4.6           22.3

         8. Nepthys caecoides              4.2  2.1            6.2

         9. Paraphoxus daboius             3.7  1.8            5.7

        10. Exogone uniformis              2-8 ' 2-9           26'8
        'Elliott, 1971.

      Although the polychaete worm Diopatra ornata is not  among the  most
common species,  its tubes are  a prominent aspect  of the bottom relief.
In the February 1976  samples from the  seep station, it constituted  34%
of the biomass,  which suggests that it  is an energetically important
species.   Its presence in a core sample also has  a demonstrable enrich-
ing  effect on the number of individuals and species  (see  Exhibit  7}.
Here, there  are significant  (p<0.01) differences  in the numbers of
individuals  and in the numbers of species  (p<0.001)  (t-test).
                                   27

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           EXHIBIT  7:   THE EFFECT OF DIOPATRA ON NUMBERS OF
               INDIVIDUALS AND SPECIES:   SEEP STATION
                             No. of Individuals, x    No. of Species,
           Samples with Dlopatra       355  42

             n = 4

           Samples without Diopatra     246  50*

             n = 6
                                                  70  5
49  6+
           'Significantly different at p = 0.01 (t-test).
           ^Significantly different at p = 0.001 (t-test).

      Bacterial Mats

      Large white mats  of  the bacterium Beggiatoa sp. are commonly seen
associated with particularly active seepage  (see Exhibit 8).  The mats
are  usually only a few millimeters or less in  thickness.  They  are often
a meter or more in diameter  and may even extend,in an irregular pattern
over larger areas of seep during calm weather.

      Beggiatoa is known as a H2S oxidizer, but  it requires aerobic con-
ditions.   These conditions appear to exist near areas of most active
seepage.   The black, anaerobic layer in the  sediment rises to the
sediment-water interface  in  these areas.  The  strong smell of H-S exists
in all our core samples;  there seems to be little doubt this compound is
abundant.
EXHIBIT 8:   BEGGIATOA MAT
  GROWING ON AREA OF ACTIVE
  SEEPAGE  [Fish in fore-
  ground  (Citharicthys sp.)
  is about  12 cm long]
                                  28

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EXHIBIT 9:  PHOTOMICROGRAPH OF
  BEGGIATOA SAMPLE FROM MAT  (150 x)
 EXHIBIT  10:   PHOTOMICROGRAPH
   OF  BEGGIATOA FILAMENT [Note
   inclusions  (400 x)]
 sediment-water interface in these areas.   The strong smell of ^S exists
 in all our core samples; there seems to be little doubt this compound is
 abundant.

      The filamentous cells of Beggiatoa (see Exhibits 9 and 10)  are
 very motile and are probably able to position themselves at the  Eh dis-
 continuity boundary.

      We have seen small nematodes within  the mat, but when we attempted
 to core a  mat the cores had such large amounts of oil that we were un-
 able to sieve them to determine if other  organisms were present.

      Other Biological Observations

      In the course of many dives in the two areas, we have the strong
 impression that the larger epibenthic organisms that are not usually
 captured in our samples are generally more abundant in the seep  area.
 These include the asteroids Pisaster brevispinous, Patiria miniata,
 Astropectin armatus, and A. californicus;  the neogastropod mollusc
 Kelletis kelletii; and the fish Citharicthys sp and Hyperprosopon
                                 29

-------
argenteum.  These last two species seemed to be attracted to our  activi-
ties, so it is difficult to judge t'heir true abundances.  Both  Hyperpro-
sopon and Astropectin were quickly drawn to the depressions left  from our
coring where they appeared to be actively feeding on exposed organisms.
DISCUSSION AND CONCLUSIONS

     The consistent differences in most measured biological parameters
in the seep area over the nonseep area seem conclusive.  The seep area
does support  an enriched community.  Most major physical parameters
 (i.e., depth, sediment type, surge, distance from shore, suspended par-
ticulates, and light penetration) appear to be about the same in the two
areas.  It is possible that some factor that is not observed or mea-
sured, such as nutrient concentrations, could be influencing these areas
unevenly; however, it seems unlikely.  Therefore, we hypothesize that
the constant presence of fresh crude oil in the sediments is responsible
for the enriching effect.

     If this  is accepted, then there are a number of further questions:

    Why are the toxic effects of the oil not more apparent?
     *
    How can crude oil enrich a benthic community?

To speculate about these questions briefly, it seems to us that either
the toxic compounds are not in solution at toxic levels in the inter-
stitial and overlying water or else the benthic organisms in the seep
area have developed the biochemical pathways to detoxify these com'-
p'bunds.  The  latter possibility seems likely with the increasing body of
evidence that points to the presence of enzyme systems capable of metab-
olizing aromatic compounds  (Payne and Penrose, 1975; Payne, 1976; Corner
et al., 1976; Lee, 1975).

     As to the enriching effect, the abundance of bacteria in associa-
tion with the active seepage suggests that petroleum hydrocarbons are
being utilized as an extra carbon source.  Although Beggiatoa has been
characterized as a chemautotroph (Zajic, 1969), it has been shown to
utilize acetate (Burton et al., 1966).  Therefore, the possibility of
these large mats utilizing some portion of the oil remains open.  The
origin of the H S may be recent or perhaps in the formation water that
could be accompanying the oil.  The increased abundance of deposit
feeders might then be attributable to increased microbial abundance.

     Clearly, this is all speculation and further observations.  Meas-
urements and experiments are needed to clarify the effects and fate of
petroleum hydrocarbons in this benthic community.
                                30

-------
    Work  in  Progress

    A number of  separate  but related efforts are currently in progress.

    The benthic  sampling  program will be continued until 18 months of
    data  are available for analysis.  This should indicate the type of
    continuing sampling effort needed to define a baseline of natural
    temporal variation.  It will also allow study of comparative pop-
    ulation  dynamics  at the two stations.

    Approximately 30  low  molecular weight hydrocarbons comprise the
    majority of  the crude seep oil.  We will be characterizing these
    and developing methods for determining these in water, sediments,
    and organisms.

    The reproductive  cycles of two populations of echinoderms thought
    to be impaired in the vicinity of the seep will be monitored.

    Attempts are being made to demonstrate acclimation phenomena by
     comparative toxicity of water of soluble fractions of crude oil to
     developmental stages of the starfish Patiria miniata.  Reproduc-
     tively active adults from the seep and a relatively pristine con-
     trol will be used.
REFERENCES

Allen, A.A. and R.S. Schlueter.  1969.  "Estimates of Surface Pollution
  Resulting from Submarine Oil Seeps at Platform "A" and Coal Oil
  Point," General Research Corporation Memorandum 1230, 43 pp.

Allen, A.A., R.S. Schlueter, and P.G. Mikolaj.   1970.  "Natural Oil
  Seepage at Coal Oil Point," Santa Barbara, California.  Science 170;
  974-977.

Buchanan, J.B. and J.M. Kain.  1971.  "Measurement of the Physical and
  Chemical Environment."  In;  N.A. Holme and A.D. Mclntyre, eds.,
  Methods for the Study of Marine Benthos, Blackwell, Oxford:  30-58.

Burton, S., R.Y. Morita, and W. Miller.  1966.   "Utilization of Acetate
  by Beggiatoa," J. Bacterial.  91;  1192-1200.

Corner, E.D.S., R.P. Harris, C.C. Kilvington, and S.C.M. O'Hara.  1976.
  "Petroleum Hydrocarbons in the Marine Food Web:  Short-Term Experi-
  ments on the Fate of Naphthalene in Calanus," J. Mar. Biol. Assoc.
  U.K.  56.:  121-133.

Elliott, J.M.  1971.  Some Methods for the Statistical Analysis of Sam-
  ples of Benthic Invertebrates.  Westmorland:   Freshwater Biological
  Assoc., 148 pp.
                                31

-------
Jones, G.F.  1969.  "The Benthic Macrofauna of the Mainland Shelf of
  Southern California," Allan Hancock Monogr. Mar. Biol.  4_:  1-219.

Lee, R.F.  1975.  "Fate of Petroleum Hydrocarbons in Marine Zooplank-
  ton."  In;  Proc. Conf. on Prevention and Control of Oil Pollution,
  American Petroleum Institute, San Francisco:  549-553.

Michael, A.D., C.R. Van Raalte, and L.S, Brown.  1976.  "Long-Term
  Effects of an Oil Spill at West Falmouth, Massachusetts."  In:  Proc.
  Conf. on Prevention and Control of Oil Pollution, American Petroleum
  Institute, San Francisco:  573-582.

Mikolaj, P.G., A.A. Allen, and R.S, Schlueter,  1972.  "Investigation
  of the Nature, Extent and Fate of Natural Oil Seepage off Southern
  California."  Presented at Offshore Technology Conference, Dallas,
  Texas.

National Academy of Sciences,  1975.  "Petroleum in the Marine Environ-
  ment."  In;  Workshop on Inputs, Fates and the Effects of Petroleum
  in the Marine Environment, Washington, D.C,, 107 pp.

Payne, J.F.  1976.  "Field Evaluation of Benzopyrene Hydroxylase Induc-
  tion as a Monitor for Marine Petroleum Pollution," Science.   191;
  945-946.

Payne, J.F. and W.R. Penrose.  1975.  "Induction of Aryl Hydrocarbon
  (benzo-[a]pyrene) Hydroxylase in Fish by Petroleum," Bull Env.
  Contarn. Toxicol.  14:  112.

Pielou, E.G.  1966.  "The Use of Information Theory in the Study of the
  Diversity of Biological Populations."  In;  Proc., Fifth Berkeley
  Symp. Mathematical Statistics and Probability.

Rohrback, E.G. and W.E. Reed.  "Evaluation of Extraction Techniques for
  Hydrocarbons in Marine Sediments."  In;  Proc. Inter. Bureau of Stan-
  dards , Santa Barbara, California, April 1975.   (In press.)

Rosen, A.A., L.R. Musgrave, and J.J. Lichtenberg.   1959.  Characteriza-
  tion of Coastal Oil Pollution by Submarine Seeps.  California State
  Water Quality Control Board, Publ. 21;  353-371.

Shannon, C.E, and W. Weaver.  1963.  The Mathematical Theory of Commu-
  nication, University of Illinois Press, Urbana,  117 pp.

Smith, J.E.  1970.  "Torrey Canyon" Pollution and Marine Life, A Report
  by the Plymouth Laboratory of the Marine Biological Association of
  the United Kingdom.   Cambridge University Press, Cambridge,  196 pp.

Straughan, D.  1971.  Biological and Oceanographical Survey of the
  Santa Barbara Channel Oil Spill 1969-1970.  Vol. I Biology and Bac-
  teriology, Hancock Foundation, Los Angeles, 426 pp.
                                32

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Straughan, D. and D.M. Lawrence.  1975.  "Investigation of Ovicell
  Hyperplasia in Bryzoans Chronically Exposed to Natural Oil  Seepage,"
  Water, Air and Soil Pollut.  5:  39-45.

Vernon, J.W. and R.A. Slater.  1963.  "Submarine Tar Mounds,  Santa
  Barbara County," California. Am. Assoc. Pet. Geol. Bull.  47;  1624-
  1627.

Wilkinson, E.R.  1971.  "California Offshore Oil and Gas Seeps,"  Cali-
  fornia, Summary of Operations, State Oil and Gas Supervisor, Tech-
  nical Papers 57:  5-28.

Zajic, J.E.  1969.  Microbial Biogeochemistry, Academic Press, New York,
  345 pp.
DISCUSSION

     Dr. Petrocelli;  In the area of chronic oil seeps, have you ob-
served any abnormal growths in any of the indigenous species similar to
some of the tumors that have been found on flat fish in that area?

     Dr. Spies:  We have not really studies these abnormal growths, but
while we were conducting the experiments fishermen mentioned organisms
with strange growths.

     Dr. Petrocelli:  Burns and Sherwood and others have reported vari-
ous types of anomalous growths and fin erosion in the fishes that con-
tact the bottom at Quellelle Point and further south.

     Dr. Spies;  Of course, there is the possibility that the solubility
of hydrocarbon is low here.  Several organisms feed on the benthic com-
munity in that area, such as cythruictes and the phenerodon, a common
surf perch.  It may be that they are acclimated to the presence of this
stuff.  They could have enzyme systems in their liver and so forth.

     From the floor:  Do you plan to use a Tellis analysis?

     Dr. Spies:  I think that the Denver ground-type of approach would
be more appropriate for a multistation type of study.  We have only two
stations.

     From the floor;  When you add the species, it should break out in a
whole bunch of different ways.  The co-occurrence of those would be the
interesting part.

     Dr. Spies:  Although we intend to get into this problem, we have
not examined appropriate methodologies for looking at -the bulk of this
data.  We are particularly interested in reproductive cycles of orga-
nisms common to both areas, recruitment, and this kind of thing.
                                 33

-------
     Dr. Davis;  Can you speculate to what degree sediment oil is
reintroduced by the water column, or does it come from geological seep-
ing?  Are there any important oils in the sediment that are coming from
the water?

     Dr. Spies:  So far, we have not made any water column measurements.
Several investigators have been measuring high molecular weight com-
pounds in the water; at a nearby station they have a level of N-alkanes
in ppb, an order of magnitude higher than most of southern California,
which has parts per trillions.   The question of the solubility of this
oil and water is open right now.
                                34

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               ACUTE AND CHRONIC STUDIES WITH WATERFOWL
                   EXPOSED TO PETROLEUM HYDROCARBONS

                             Michael P. Dieter*
     Our requirements for energy and economic independence will rest
largely on petroleum hydrocarbons for several decades.  This recognition
has resulted in concerted efforts to find, obtain, process, and deliver
oil at an ever-increasing rate.  Coupled with this phenomena is our
nation's awareness and dedication to the maintenance of ecological bal-
ance.  Many government agencies have been assigned the task of ensuring
our wildlife heritage during the removal of nonliving natural resources.
We represent a small part of this effort:  an examination of the ecolog-
ical and physiological/toxicological effects of oil on birds.  Our stud-
ies, which encompass numerous aspects of avian biology, are proceeding
on a broad front as fast as possible in an attempt to answer one basic
question:  do sublethal concentrations of petroleum hydrocarbons pose a
significant threat to bird populations?

     The effects of petroleum hydrocarbons on aquatic birds have been
the subject of two recent symposia  (Dieter, 1976; Szaro, 1977).  These
reviews pointed out that the base of scientific information was scant
and needed expansion.  Nevertheless, the available evidence was alarm-
ing and indicated the potential damage that petroleum hydrocarbons might
inflict on susceptible avian populations.

     The research program at Patuxent has been evaluating the biological
effects of petroleum hydrocarbons on all stages of the waterfowl life
cycle.  These studies include:  (1)  the hatchability of eggs exposed to
oils;  (2) the development of ducklings fed oil from the day of hatch;
and  (3) reproduction in adult waterfowl fed oil.  The third study is
complemented by another where adult waterfowl were fed a chemically
defined mixture representative of the concentration and type of aromatic
compounds present in oil.  In addition, experiments have been initiated
to evaluate biological responses to petroleum hydrocarbons incorporated
via the food chain.  Analytical development is proceeding in concert
with all of these biological studies.
*    Fish and Wildlife Service, Patuxent Wildlife Research Center,
Laurel, Maryland 20811.
                                35

-------
     We have repeatedly demonstrated that yJl amounts of crude or refined
petroleum hydrocarbons result in high mortality of developing embryos.
Exhibit 1 shows that 5 \il of South Louisiana crude oil, Kuwait crude
oil, or No. 2 fuel oil, applied to fertile eggs at 8 days of incubation,
caused 76-98% mortality.  We have also concluded that mortality is a
result of toxicity and not oxygen deprivation, and that an alkane mix-
ture tested was nontoxic, implicating the aromatic fraction or other
constituents of crude oil as the toxic components.

   EXHIBIT  1:  EMBRYOTOXICITY FROM EGG OILING* WITH DIFFERENT PETROLEUM
                             PRODUCTS  (N=50)
Percent Mortality after Treatment with:
Treatment
None
Propylene
glycol (50 y)
Alkane
mixture (50 y)
Oil (1 y)
Oil (5 y)
Oil (10 y)
Oil (20 y)
South La.
crude oil
8
6
4
38**
98**
98**
100**
Kuwait
crude oil
8
-
-
28**
76**
84**
94**
No. 2
fuel oil
12
-
-
36**
82**
90**
100**
 Note:  Data derived from the studies of .Szaro,  Albers,  and Coon (1977)

 *    Fertile mallard eggs treated externally at day 8 of development.
 t    See Exhibit 6 for components of alkane mixture.
 **   Significantly different from controls, P < 0.01.
      The embryotoxicity of oils was not confined to the mallard.   Eider
 eggs collected from the Maine seacoast were externally oiled with No.  2
 fuel oil resulting in 31% mortality (see Exhibit 2).   Eider embryos
 appeared to be 1/20 as sensitive as mallard embryos to external egg oil-
 ing.  Some of the species difference can be ascribed to different dose-
 weight relationships, as the eider embryos are twice the size of mallard
 embryos.  Age-dependent sensitivity to petroleum hydrocarbons is another
I factor that was responsible (see Exhibit 3).   Embryos that were at ear-
 lier stages of development were much more sensitive to egg oiling than
 later stages.  This was supported by another series of experiments shown
 in Exhibit 4.
                                 36

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      EXHIBIT 2:  EMBRYOTOXICITY  IN  OILED  COMMON EIDER EGGS*  (N=48)
            Treatment                     Percent Mortality

            None                                 4
            Propylene glycol  (50 y)             4
            Oil  (5 yfc)                           8
            Oil  (20 y)                          31^
	-/	,	,	,	
Note:  Data derived from the studies of Szaro and Albers  (1977).
*    Fertile eider eggs of various developmental stages treated  exter-
nally with No.  2  fuel oil.
t    Significantly different from controls, P <  0.01.
   EXHIBIT 3:  AGE-DEPENDENT EMBRYOTOXICITY* TO OIL IN COMMON EIDER EGGS
             No.  2  Fuel  Oil                             Age in Days
                 (20 y)                  N            (Mean  S.E.)
             Dead                       15              4.3   1.4


             Survivors                  33             16.1   0.7

 Note:   Data derived from the studies  of Szaro and Albers  (1977).
 *     Significantly different ages,  P  <  0.05.
                                 37

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 EXHIBIT 4:  EMBRYOTOXICITY FROM EGG OILING* AT DIFFERENT DEVELOPMENTAL
                            STAGES  (N=50)


                           Percent Mortality after  Treatment with:
 Age  of  Treated
 Embryo  (days)            South  Louisiana Crude Oil      No. 2 Fuel Oil
1"
Controls
2
6
10
14
18
22

0
100**
97**
92**
22**
12**
5

20
87**
67**
32**
17
20
7
 Note:   Data derived  from Albers  (1977).
 *    Fertile mallard eggs treated externally with 5 y of oil.
 t    Mortality of untreated control eggs checked by candling at each
 interval.

 **   Significantly different from controls, P < 0.05.
     Mortality  from petroleum hydrocarbon application was clearly age
dependent;  5  y applied on or before 10 days of development resulted in
92% or greater  mortality from South Louisiana crude oil and 32-87% mor-
tality from No. 2 fuel oil  (see Exhibit 4).  Treatment of eggs on or
beyond day  14 resulted in much less mortality.  This line of investiga-
tion will be  continued with eggs from different species and with other
crude oil products, and will be extended by conducting field studies
with eggs oiled in marked nests and outdoor pen studies with oiled
waterponds  and  naturally incubating hens.

     We also  found that hens from paired mallards fed 2.5% South Loui-
siana crude oil laid fewer eggs than pairs fed 0.25%, a mixture of 1%
paraffins, or untreated control pairs.   Egg production for 30 days
dropped from an average of 25 eggs/hen in untreated to 11 eggs/hen in
those fed 2.5% South Louisiana crude oil.  Our contract investigator
has reported  similar results with mallards fed 1%, 3%, or 5% South
Louisiana crude oil (Holmes, personal communication).  He reported that
hens fed 3% oil ceased laying, and that those fed 1% oil laid one-third
as many eggs as controls.   It is evident that, during the reproductive
season,  petroleum hydrocarbons could pose a threat to waterfowl popula-
tions by interfering with the production and viability of eggs.
                                38

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  EXHIBIT 5:  MORPHOLOGICAL AND BIOCHEMICAL  RESPONSES  IN  MALLARD
DUCKLINGS FED SOUTH LOUISIANA CRUDE OIL  FROM HATCH  TO  8 WEEKS  OF AGE
                        (MEANS  S.E., N=10)
Cone entration
Oil Fed
(ppm)
0
250
2500
25000
50000
Body Weight
g
1119
1127
1109
1069
913
      Morphology
  Liver Weight
     g      % BW
34.5  1.5   3.2
33.0  1.9   3.0
31.5  1.7   2.9
5L. 5 1.9   4.8*
69.7  4.7   7.4*
                                                     Spleen Weight
                                                       g       % BW
                                                  0.81  0.06  0.074
                                                  0.84  0.12  0.067
                                                  0.64  0.04  0.058*
                                                  0.39  0.06  0.037*
                                                  0.30  0.03  0.032*
                                    Biochemistry
                     Plasma Enzyme Activity
                                                t
                Alanine
            aminotransferase
               (from liver)
               milliunits
0
250
2500
25000
50000
11.6 
12.7 
23.5 
19.0 
16.3 
1.0
2.1
3.3*
1.6*
1.4*
12.5 +
23.1 
19.0 
18.8 
20.7 
0.6
1.2*
1.8*
1.1*
1.5*
       Ornithine         a-hydroxybutyric
  Carbamyl transferase  acid dehydrogenase
     (from kidney)          (from heart)
         units              milliunits
                            96.4  -9.0
                           112.3   8.9
                            84.6   7.8
                            77.8   7.7
                            86.3  12.5
Note:  Data derived from Szaro  (unpublished data, 1977).
*    Significantly different from controls, P < 0.05.
t    Additional plasma enzymes measured that showed no significant
change included aspartate aminotransferase, lactate dehydrogenase, and
cholinesterase.
     Duckling growth was impaired when diets containing 5% South Louisi-
ana crude oil were ingested from hatching until 8 weeks of age (see
Exhibit 5).  More importantly, in these ducks, as well as those fed 2.5%
South Louisiana crude oil, flight feathers failed to develop normally.
Liver hypertrophy and splenic atrophy were evidence of the pathological
effects of the oil; biochemical lesions that occurred included elevation
                                39

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of plasma alanine aminotransferase and ornithine carbamyl transferase
activity.  These plasma enzymes appear in the circulation as a result
of liver and kidney damage.

     The same enzymes are being measured in studies with adult ducks
fed South Louisiana crude oil or representative aromatic compounds of
this crude oil.  These variables have thus far failed to reveal dys-
function in adult ducks fed up to 2.5% South Louisiana crude oil, or
in those fed the aromatic mixtures.

     More sophisticated measurements of liver function were performed
with the drakes fed the aromatic mixture.  The ability of the liver to
remove toxic substances from the circulation was followed by measuring
the change in the clearance rate of injected indocyanine green dye, a
compound that is metabolized entirely by the liver.  The ingestion of
the 4000 ppm aromatic mixture caused a compensatory response to the
toxicants reflected by a significant elevation of plasma clearance
rates above control levels (see Exhibit 6).   The data suggest that adult
waterfowl may be able to adapt and tolerate higher concentrations of
petroleum hydrocarbons than ducklings.

     Thus, our present evidence indicates that all phases of the repro-
ductive sequence  (i.e., fertilization, laying, and embryonic and duck-
ling development) are highly sensitive to petroleum hydrocarbons.
Hopefully, extensive measures could be undertaken to prevent oil expo-
sure during this critical period.

     In other studies, waterfowl were fed various invertebrates that
had been exposed to petroleum hydrocarbons to determine the nature and
extent of the products accumulated in their tissues.  Crayfish have been
been selected as food items that are readily accepted by mallard ducks.
A radiolabeled aromatic compound is used as a marker to follow the oil
through the food chain.

     Along with the biological work, a concerted effort has been made to
develop analytical capability for petroleum hydrocarbons.  This poses a
special challenge because of the overwhelming number of chemical com-
pounds present in one sample, and the fact that this array is unique
for different oils and even for different batches of the same oil.

     The first problem addressed was the demonstration of the accumula-
tion of petroleum hydrocarbons in avian tissues.   High-resolution
gas chromatography and mass spectrometry were used to identify the
petroleum-derived, saturated hydrocarbons in tissues of oil-dosed ducks
(Lawler, e_t_ al^., 1977).  The distribution pattern of n-alkanes, the
homologous series of petroleum hydrocarbons, and the n-Ci7/pristane
ratio were used to provide evidence that petroleum hydrocarbons had
accumulated in the tissues.  Selective uptake and/or metabolism had
apparently occurred since tissue levels of individual saturated petro-
leum hydrocarbons differed from the relative amounts present in South
Louisiana crude oil.  No accurate quantitation of saturated petro-
leum hydrocarbons was possible, but relative accumulation by tissues

                                40

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EXHIBIT 6:  LIVER FUNCTION IN MALLARD DRAKES FED AROMATIC MIXTURES*
       REPRESENTATIVE OF THOSE IN SOUTH LOUISIANA CRUDE OIL
                        (MEANS  S.E., N=12)
                                                Months

                  Pretreatment         13                5

Treatment               Plasma Clearance Rate  (ml/min/kg body weight)

Control           13.7  1.1      13.5  0.8     13.5  0.7  13.1  0.7

Alkane mix,
10,000 ppm        14.0 + 1.1      14.7  0.7     14.9  0.7  13.1 + 0.7

Aromatic mix,
400 ppm           13.4  1.0      15.5  0.8     15.2  0.7  14.7  0.5

Aromatic mix,
4000 ppm          14.1  1.0      17.0  1.4     18.5  1.0  18.5  1.2
Note:  Data derived from Patton  (unpublished data, 1977).

*    The aromatic mixture consists of an equimolar ratio of 10 compounds
(ethyl benzene, 1,2,3,4-tetrahydronaphthalene, dimethylnaphthalene, 2,
3-trimethylindolenine, acenaphthylene, acenaphthene, phenanthrene, 2-
methylbenzothiazole, dibenzothiophene, and 2,6-dimethylquinoline) dis-
solved in 1% alkanes  (equimolar mixture of tridecane, pentadecane, hexa-
decane, heptadecane, octadecane, nonadecane, 2,2,4,6,6-pentamethyl hep-
tane , 2,2,4,4,6,8,8-heptamethylnonane, 2,6,10,14-tetramethylpentadecane,
and decahydronaphthalene) in the feed.
t    Significantly different from controls, P < 0.05.
was:  skin > uropygial gland > breast muscle > heart muscle > liver >
blood.  No saturated petroleum hydrocarbons were detectable in the brain
and only trace amounts in the blood.

     The unsaturated petroleum hydrocarbons (aromatic compounds) are now
being measured in the same tissue extracts.  The tissue accumulation of
aromatic hydrocarbons was similar to that of the alkanes, except that
detectable quantities of aromatics were present in the brain.

     Our analytical efforts favor the aromatic fraction because the bio-
logical evidence indicates that the aromatic compounds in petroleum
hydrocarbons are toxic.  We are streamlining present analytical methods
and are selecting marker compounds in petroleum hydrocarbons to serve as
                                41

-------
references for quantitation.  Pending completion of these tasks, tissues
from mallard ducklings fed South Louisiana crude oil are scheduled for
petroleum hydrocarbon analysis.  Other material that will soon become
available for analysis includes tissues from adult mallards fed either
South Louisiana crude oil or an aromatic mixture representative of this
oil.
REFERENCES

Dieter, M.P.  1976.  "The Effects of Petroleum Hydrocarbons on Aquatic
  Birds."  In;  Proc, Symp. Sources, Effects, and Sinks of Hydro-
  carbons in the Aquatic Environment.  American Institute of Biological
  Sciences, Washington, D.C.:  437-446.

Lawler, G.C., W.A. Loong, and J,L. Laseter.  1977.  "The Accumulation of
  Saturated Hydrocarbons in Tissues of Petroleum Exposed Mallard Ducks
   (Anas platyrhynchos)."   (In press.)

Szaro, R.C.  1977.  "The Effects of Petroleum on Birds."  Iii:  Trans.
  of the North American Wildlife and Natural Resources Conference.
  Atlanta, Georgia.   (In press.)
                   *
Szaro, R.C. and P.H. Albers.  1977.  "Effects of External Applications
  of Oil in Eider Eggs."  In:  Proc. Symp. Fate and Effects of Petro-
  leum Hydrocarbons in Marine Ecosystems and Organisms.  Seattle,
  Washington.   (In press.)
 DISCUSSION

      Dr.  Anderson;   Have  you  tried the oiling of the eggs with weathered
 crude,  because of the  aromatic decrease?

      Dr.  Dieter:  In our  current series of experiments, we are using
 Alaskan crude  oil and  weathered and unweathered oils.  We are also going
 to  use  some water-soluble fractions to investigate whether we can find
 a decrease in  embryo toxicity.  50,000 parts commutes to a very high
 physiological  or  pharmacological dose.

      Dr.  McErlean:   Do you have any feeling of the effects of an oil
 spill?

      Dr.  Dieter:  I  would speculate that we would be looking at re-
 sponses from ducks at  the level of 250 or perhaps 2,500 ppm; 50,000 ppm
 is  definitely  a pharmacological level.  We wished to bracket the levels
 where we  might see an  effect, and then work down to make sure that we
 did not miss anything.  In these chronically fed birds, 250 ppm is
 approximately  2.5 mg a day ingested per duck; in the case of ducklings
 it  is over an  8-week period.
                                42

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     Dr. Engel:  What was the label compound used?

     Dr. Dieter:  We began with C-14 naphthalene and will try other
labels.  So far, we have not performed any metabolite studies.  We
have calculated the DPMs and the rough ppm naphthalene that can be
surmised from the disintegrations in the tissue.  This is over 1 ppm
in the gall bladder of the ducks fed the radio-labeled crawfish, a
two-fold and then a three-fold increase in the radio-labeled compound
in the fat, a steady increase in the kidney, and a slower increase in
the liver.  From this food chain study, the gall bladder is the highest
accumulator so far, although that is probably all metabolite; the fat
is the next highest, and then the kidney and the liver.

     Dr. Kunhalt:  When did you check whether the eggs were dead or did
you count the hatchlings or surviving ducks?

     Dr. Dieter;  The majority mortality resulting from the external
exposure of the ml amount of oil occurs within 6 days after exposure.
The eggs are checked every other day for mortality by candling.  If
they appear to be dead, then they are opened to verify this conclusion.

     Dr. Kunhalt;  I am not familiar with the development stages in
ducks.  You mentioned, first, the application of 8 days and showed the
mortalities.  What stage is that?

     Dr. Dieter:  It is a full embryo.  For the blastula stage it is
below  the first day.
                                43

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       ACCUMULATION AND BIOLOGICAL EFFECTS OF COPPER AND CADMIUM
                          ON MARINE SHELLFISH

                    David W. Engel* and Bruce A. Fowler
     In recent years, the interface between research related to human
health and ecological research has become much less distinct.  The in-
terplay between these two areas of research has become more relevant
because of our expanding technology and greater demand for electric
power.  As a result of our increased demand for energy, there has been
an increase in trace metals entering the waters of estuaries and oceans.
Some of these metals come directly from energy production  (i.e., the
burning of fossil fuels), from the cooling process  (i.e., corrosion
products from condenser tubes), or from industrial and municipal waste.
These metals can accumulate in marine fish and shellfish, which may
alter their capacity to withstand natural environmental stresses.  In
addition, these organisms may become food for human consumption, and
the accumulated metals will enter the human diet.

     The research that we are conducting concerns the effect of copper
and cadmium on the metabolism of shellfish.   Copper may enter the envi-
ronment from power plants and industrial sources, while cadmium enters
primarily from industrial waste and sewage effluents.

     In most of the pharmacological investigations concerned with the
toxic effects of metals on mammals, the metal is added to the diet as a
mineral salt.  However, most dietary metals are present as organocom-
plexes, and these different chemical species may drastically affect
their toxicological and pharmacological properties.  In our laboratory,
we have demonstrated that the various chemical species of copper can
control the toxic effects of the metal on marine fish eggs and larvae
and algae (Engel ejt al^., 1976; Sunda and Guillard, 1976;  Sunda,  1975).
To approximate acute pollution situations, shellfish are being labeled
with metals  (copper and cadmium) over extended periods of time, and the
labeled meat will be used in the production of a  "toxic" rat diet that
represents more nearly a natural situation.
*    National Marine Fisheries Service, Beaufort Laboratory, Beaufort,
North Carolina 28516.
t    The National Institute of Environmental Health Sciences, Research
Triangle Park, North Carolina 27709.
                                45

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     The two objectives of our research are:

1.   Label shellfish with biologically incorporated copper  (Cu) and
     cadmium  (Cd) that will be used in the preparation of rat diets
     for pharmacokinetic and toxicity investigations

2.   Determine physiological and fine structural changes in marine
     shellfish caused by copper and cadmium.
     Experimental Design

     The  flowing water system, which was constructed for metal  expo-
 sures, is  similar to the system being used for exposure of organisms  to
 pesticides at the Environmental Protection Agency, Gulf Breeze  Pesti-
 cide Laboratory  (Banner, e_t a^. , 1975).  The primary difference between
 the two systems is that we use ambient sea water.

     Sea  water is pumped through a coarse filter into the 640  recir-
 culating  temperature control tank  (Living Stream Unit),* where  it is
 filtered  again through activated charcoal, and heated or cooled to main-
 tain a constant temperature  (see Exhibit 1).  Water is then pumped from
 the temperature control tank up to a 95 A polyethylene reservoir, which
 functions  as a constant head tank.

     The  water-toxicant distribution system, experimental tanks, flow
 meters, pipes, tubing, and syringe pumps are supported by a heavy-duty
 wood frame with two 3 in. deep trays.  The insides of the trays are
 fiberglass lined, and each tray is connected to a drain.  Two and five
 38 H experimental tanks can be accommodated on the upper and lower lev-
 els, respectively.  Gilmont flowmeters were used to monitor the flow  of
 diluent water from the manifold to the tanks, and the flow rates may  be
 set between 5 mi and 350 m per minute.

     Sage  syringe pumps are used to inject the toxicant at rates that
 can be varied from 4 x 10~  m to 140 mi/min.  Each pump holds  from one
 to three  glass or plastic syringes, depending on size.  Thus, either  one
 100 m syringe, two 50 m syringes, or any three syringes up to 30 m
 can be used.  Toxicant and diluent water flow separately through appro-
 priate plastic tubings to mix in a glass funnel before flowing  into the
 experimental tanks.  Test water flows out of the experimental tanks
 through glass T-joints cemented in the side ends of the tanks.

     Using the flowing water exposure system described above, the oys-
 ters were  labeled separately with copper or cadmium.  The oysters
*    The use of trade names does not constitute an endorsement of these
products by National Oceanographic and Atmospheric Administration
(NOAA) or the National Marine Fisheries Service  (NMFS).
                                46

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         SVRINQC PUMP
   WATER MANIFOLD
        n
    TO OKM.IN fttLO
                            O	Q

              v

                                                               COMPRESSOR
                                        TEMPERATURE CONTROL TANK
EXHIBIT 1:  A FLOWING WATER BIOASSAY  SYSTEM, WHICH IS  BEING USED TO
  DETERMINE THE TOXIC EFFECTS OF METALS ON MARINE  FISH AND SHELLFISH
   (Arrows indicate the pattern of water flow in  the system.)
were exposed foe 14 days at 0.1 ppm  concentration  of  either copper or
cadmium.  During the exposure period, the oysters  were  fed  twice  a week
with mixed algal suspensions, which  were spiked with  0.1 ppm of copper
or cadmium.  At the end of the exposure period, the meats were removed,
freeze dried, and used in preparation of rat diets for  mammalian  phar-
macokinetic studies.  The levels of  cadmium in the oysters  ranged from
500 ppm to 1000 ppm dry weight, and  the mean concentration  of copper
was 360 ppm.
EFFECTS OF COPPER ON SHELLFISH

     To determine the physiological effects of copper on shellfish, we
measured  shellfish  gill  tissue  respiration  rates.   Four  species  of
lamellibranch mollusks were used in the determinations:  the American
oyster, Crassostrea virginica;  the hard clam, Mercenaria mercenaria;
the bay scallop, Argopecten irradians; and  the ribbed mussel,  Modiolus
demissus.  All animals were collected in the vicinity of our laboratory
at Beaufort, North Carolina, and maintained in the  laboratory  in flow-
ing sea water.

     The copper solutions used  in the experiments were all buffered with
5 mM "Tris" in 30 ppt salinity  Sargasso Sea water at pH 8.0 +  0.05.
                                47

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                                                            5
The total copper concentrations of the solutions were 2 x 10  M,
5 x 10~5M, 1 x 10~4M, and 5 x 10~4M with cupric ion concentrations of
ICT^'^M, 1Q~8.3M^ io~8-01f4f and 10~^-^-^M.  The control seawater con-
tained "Tris" and a copper concentration of <10~8M (Chester and Stoner,
1974), which would give a cupric ion concentration of <10~10M.  The
cupric ion concentrations were derived from the apparent stability con-
stants for copper in seawater (Sunda, 1975).  The organic buffer system
concept used with copper will reduce the complexation of copper by nat-
ural organics in the exposure media, because of the high ratio of total
copper to free cupric ion  (10,000 to 1).  This method allows the predic-
tion of a specific chemical species of copper as the toxic agent (free
cupric ion) and results in reduced variability between and within exper-
iments .

     The measurements of tissue respiration were made with a Gilson
differential respirometer using standard techniques.   All measurements
are reported as y of oxygen consumed per mg dry weight per hr.

     The excised gill tissue from the four species of mollusks exhibited
species-specific responses to copper.  The respiration rates of scallop
and clam tissue increased significantly with increased cupric ion con-
centrations.  Gill tissue from the mussel, however, had a relatively
smaller increase in respiration than the scallop or clam tissue, and the
respiration rates of oyster gill tissue were not significantly affected
by the presence of copper  (see Exhibit 2).  The response of the clam
gill to copper was roughly linear, and the variation within each concen-
tration was quite low.  Scallop tissue,  however, had exceedingly vari-
able respiration rates at cupric ion concentrations above 10" M.   The
primary source of the variation was the copper-induced breakdown of the
tissues, which resulted in variable tissue weights.  It is interesting
to note that the scallop was also the most sensitive to copper in pre-
liminary toxicity tests.

     When the respiration data were normalized to the control respira-
tion rates, a clearer picture of species similarities and differences
relative to the effect of copper was demonstrated  (see Exhibit 3).
These responses fall into two categories:  the scallop and clam tissue
had increased respiration with increased copper concentration, while the
mussel and oyster gill had increased respiration at cupric ion concen-
trations between 10~8-7M and 10~8M followed by decreased respiration.
These differences may be related to the salinity tolerance of the spe-
cies, with mussels and oysters more euryhaline than either clams or
scallops.  The observed increase in respiration may be related to
copper-induced changes in membrane permeability to ions (Na+ and K+) as
has been demonstrated with algae cells  (McBrien and Hassall, 1965).
There also may be copper-mediated changes in mitochrondrial membranes
causing uncoupling of the electron transport systems.  This possibility
is currently under investigation as part of this research program.  The
observed effects of cupric ion on molluscan gill tissue are in the same
range as toxic effects of free cupric ion on spot egg and larvae and
unicellular algae  (Engel ejt al_. , 1976; Sunda and Guillard, 1976).
                                48

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                                    3.0
                                    2.0
EXHIBIT 2:  CHANGES  IN  MOLLUSCAN
  GILL TISSUE RESPIRATION RATE AS   1.0
  A FUNCTION OF THE  CUPRIC ION CON-
  CENTRATION    (Concentration of
  cupric ion given both in molar
  units and as ppb.   Vertical bars
  indicate one standard error of
  the mean respiration  rate,  and
  the horizontal bars indicate the
  range in cupric ion concentra-       Q06   Q6
  tion.  There are 10 replicates
  for each mean value.)
                                                       Scallop
        lop     T.--
        SXT
                                         ,10
10
                                                       10'8
                                                    Cu2+(M)
                                                        .6
                                                    Cu'Mppb)
               10
EXHIBIT 3:  NORMALIZED  RESPIRA-
  TION RATES OF MOLLUSCAN GILL
  TISSUE AS A FUNCTION  OF CUPRIC
  ION CONCENTRATION
                                   2.0

                                   1.8

                                   1.6

                                   1.4

                                   1.2

                                   1.0
Scallop  -
                                        1010   1C9
       10
                                                                      10
                                                  Cu2+(M)
                                        .006   .06
       .6
                                 49

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     In another series of experiments, we exposed  gills  from clams and
oysters to two different salinities and two copper concentrations  and
recorded respiration rates.  The salinities used were 30 ppt and 10 ppt
and cupric ion concentrations were <10~10M for the controls  and 10  ' _M
for the experimental.  Again, the respiration rate was measured with a
Gilson respirometer.

     The respiration rates of excised clam and oyster gill were affected
differentially by salinity and cupric ion  (see Exhibit 4).   Although
salinity had a significant effect upon the respiration of clam gill,  it
had no effect upon  oyster gill.  The data on the effects of  salinity  on
gill respiration agree favorably with that of Van  Winkle et  al.  (1968),
who demonstrated similar relationships with oyster and clam  gill.
   1.0 -
           EXHIBIT 4:  HISTOGRAM OF CLAM AND OYSTER GILL TISSUE
             RESPIRATION RATES AS A FUNCTION OF SALINITY AND
             TOTAL COPPER CONCENTRATION  (Cupric ion concentra-
             tion 10~8M).  CONTROL COPPER CONCENTRATION WAS <10~9M
              (Cupric ion concentration <10~1:1-M) .   (Dotted area
             above and below horizontal line is one standard error.)


     On the other hand, copper influenced gill tissue respiration in
both species.  An ANOVA of the clam data revealed that, while salin-
ity and copper both influenced respiration,  there was no signifi-
cant interaction between the two factors (<5 = 0.05).  Although the
                                50

-------
interaction was not significant, it approached significance and sug-
gests that the influence of copper was greater under high salinity con-
ditions.  The ANOVA of the oyster gill respiration data demonstrated
that the effect of copper on gill respiration was significant, and the
increased respiration was not dependent upon salinity  (i.e., there was
no salinity x copper interaction).  The disparity between these data on
oyster gill and those from the previous investigation may have been the
result of using smaller gill tissue samples  (i.e., 50%), and reduced
variability in the data.  Furthermore, the second experiment was run in
a different season of the year.

     Because experiments with acute exposures of excised gill tissue to
different cupric ion concentrations have demonstrated that cupric ion
or copper in the exposure media affects the respiration of excised mol-
luscan gill tissue, experiments were conducted to determine if accumu-
lated copper would significantly affect gill respiration.  Oysters were
used because of our experience in holding them during extended expo-
sures to metals.  We designed a series of experiments to test the ef-
fects of accumulated copper upon the respiration rate of oyster gill
tissue.  The oysters were exposed in a flowing sea water system con-
taining copper at 0.1 ppm as copper chloride; the exposure period was
14 days, with samples taken at 1, 3, 7, and 14 days.  After the animals
were removed from the exposure tanks, they were allowed to flush in
clear water for 24 hr prior to dissection.  Tissue was sampled for mea-
surement of respiration, determination of total copper, and fixed for
electron microscopy at day 0, 1, 3, 7, and 14.

     The accumulated copper in the oyster gill tissue significantly af-
fected the tissue respiration rate by the fourteenth day (see Exhibit
5).  There was a steady increase in the respiration rates and copper
concentrations of the treated oyster tissue relative to the control tis-
sue from the first through fourteenth day.  While the increase in res-
piration was significant for the copper-treated oysters, there was no
significant change in the control tissue respiration over the same time
period.  It should be noted that by 14 days the remaining oysters were
in poor condition, presumably from accumulated copper.  Therefore, the
increased respiration may be associated with the lethal effect of the
accumulated copper.

     In addition, ultrastructural examination of oyster gill epithelial
tissue was conducted.  Preliminary results indicate that there are some
dense inclusions present in the tissue, but, at this time,  it is not
possible to correlate either the densities or numbers of inclusions
with copper exposure.  We also plan to conduct elemental analysis of
the inclusions with the electron probe.
                                51

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           1.5
           1.0
        CD
        E
        \
        CM
        o
        Z
        O
Si 0.5
UJ
01
            .Control Resp.
            . Cu Treated Resp
            .. Tissue Cu Cone.
              	'
            f
          I  j   j	i
                            i   i   i   i
                                       I   I
4    6     8    10

 DAYS OF EXPOSURE
                                               12
                                            14
                                 .5
                                                  .4
                                                          .3
                                    I
                                    O
                                                     a
                                                      0>
                                                      E
                                                     u
                                                  .2  E
                                                     O)
                                                     a.

                                                  .1
        EXHIBIT 5:  THE EFFECTS OF TREATMENT WITH 0.1 PPM COPPER
          ON THE RESPIRATION RATES OF EXCISED OYSTER GILL TISSUE
          AS A FUNCTION OF TIME OF EXPOSURE, AND THE ACCUMULATION
          OF COPPER IN GILL TISSUE AS A FUNCTION OF TIME  (Vertical
          bars indicate one standard error of the mean for 10 rep-
          licates in the respiration determinations and 5 replicates
          in the tissue copper concentrations.)
EFFECTS  OF  CADMIUM ON SHELLFISH

      The data on  the  toxicity  of  cadmium to  aquatic species indicate
that  fresh  water  species  are somewhat more sensitive to cadmium than
marine species  (Negilski,  1976? Eisler,  1971),  and that there is a
salinity dependence of cadmium toxicity  on euryhaline organisms (Jones,
1975; von Westernhagen and Dethlefsen, 1975),   To demonstrate how salin-
ity affects cadmium toxicity and  availability,  an investigation was con-
ducted using the  grass shrimp,  Palaemonetes  pugio,  as the test organism.
To test  for toxicity,  the  shrimp  were exposed to  1.0,  0.5,  0.25, 0.125
ppm and  <0.1 ppb  of cadmium at salinities of 4, 8,  16, and 28 /oo.
Exposures were made in 4   polyethylene  tubs for  96 hr.

     Measurements  of  cadmium availability and uptake were made using
115mcd at salinities  similar to those used for  the toxicity tests.
Using liquid scintillation counting  techniques, we measured the uptake
of the radionuclide by grass shrimp  every 24 hr for a period of 96 hr.

     The results of the cadmium exposures indicated that the toxicity
of the metal was salinity  dependent  (see Exhibit  6).   The greatest tox-
icity was at the lowest salinity, which  indicates that osmoregulatory
                                 52

-------
damage may be involved.  Similar data have been  generated for marine
and estuarine isopods  (Jones,  1975)  and  for marine  fish eggs  (von
Westernhagen et^ a^. , 1974).  Thus, the salinity  dependence of cadmium
toxicity appears to be rather  universal.
                                    SALINITY (%0)

          EXHIBIT 6:  A THREE-DIMENSIONAL PLOT OF GRASS SHRIMP
            96 *HR SURVIVAL AT FOUR DIFFERENT SALINITIES AND FIVE
            DIFFERENT CADMIUM CONCENTRATIONS (Twenty shrimp were
            used in each treatment.)

      When the experiment was repeated and survival of shrimp plotted
 against cadmium ion concentration rather than total cadmium concentra-
 tion,  the direct correlation between  survival and salinity was  modified
 (see  Exhibit 7).   Survival was  dependent upon measured cadmium  activity,
 which,  in turn,  was dependent upon salinity.   The observed change in
 cadmium toxicity with salinity  can be explained through the research of
 Sunda and coworkers in our laboratory,  who have demonstrated that cad-
 mium  toxicity is dependent upon the chloride concentration of the water
 (i.e.,  salinity), which is from the inorganic complexation of cadmium
 by  chloride.  The cadmium-chloride complex is independent of pH and is
 not affected by dissolved organic matter in the water.   Thus, the
 cadmium-chloride complex is less toxic to grass shrimp than free ionic
 cadmium.

      The  uptake curves generated in these experiments also indicated
 that  the  bioavailability of cadmium is controlled by the chloride con-
 centration of the water (see Exhibit  8).   For shrimp maintained at 5,
 10, and 20  /oo,  the uptake curves were roughly linear for 96 hr, while
 the data  at 30   /oo were somewhat more erratic.   The rate of uptake was
 greatest  at 5  /oo and slowest  at 20  and 30  /oo.  The fact that the
                                53

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z
UJ


a.
    100
     80
     60
     40
     20
                         \
                                 Salinity (%)


                                 * 4.8

                                  8.4

                                  16.5

                                  20.0

                                 * 28.9
                                \.
              10
                                   10
                        Cd24(M)
              11.2
                                   112.4
                       Cd2t(ppb)
 EXHIBIT 7:  THE  96  HR SURVIVAL OF  GRASS SHRIMP AS  A

   FUNCTION OF THE MEASURED FREE CADMIUM ION CONCEN-

   TRATION IN THE MEDIUM
     5.0


 o


 X   4.0
 -a

 u>

 E
 ^

 S
 a.
     3.0
2.0
     1.0
                  24
                        48


                       HOURS
72
96
  EXHIBIT 8:  THE  UPTAKE OF U5mcd  BY GRASS SHRIMP AT

    FOUR DIFFERENT SALINITIES FOR 96 HR  (Each point is

    the mean of  five individuals with one standard error.)
                             54

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curves did not begin to level off by 96 hr .simply indicates that equi-
librium between the grass shrimp and 11^inCd in the water had not been
obtained.  To determine if salinity also affects cadmium loss, experi-
ments are planned to test the rate of loss of     Cd from shrimp that
have been exposed to cadmium at different salinities.

     We attempted to measure the effects of cadmium on molluscan gill
tissue  (i.e., clams and oysters) in a series of experiments similar to
the copper series  (i.e., both acute exposure and accumulated metal).
All experiments were conducted at 30 /oo and at a series of concentra-
tions of cadmium ranging from 1.0 ppm to 0.125 ppm, but the results were
negative.  The lack of any effect on the respiration rates of molluscan
gill tissue is probably a result of the complexation of cadmium by
chloride, since all the experiments were carried out at 30 to 35 /oo
salinity.  We can speculate that the complexation of cadmium at high
salinity makes it improbable that much, if any, cadmium could pass
across  a cell membrane.  At lower salinities, 15  /oo and below, our
data indicate that this does not appear to be true, and we are currently
conducting experiments to test the hypothesis.

     The data presented in this report, along with corollary information
currently being generated in our laboratory, indicate that the chemical
species of an* element determines its pharmacological and toxicological
effects on aquatic organisms.  This has been demonstrated for two ele-
ments,  i.e., copper and cadmium.  The toxicity of copper to algae and
fish eggs and larvae  (Sunda and Guillard, 1976; Engel et al., 1976) was
shown to be related to the "free" cupric ion, and that the complexation
of copper is by organic chelators and the degree of complexation is pH
dependent.  Cadmium, on the other hand, is complexed by chloride, an
inorganic ligand, and the degree of complexation is independent of pH
arid the dissolved organic load in the water  (Sunda, unpublished data).
Thus, it seems reasonable for us to continue our research on the effects
of metals, copper, cadmium, and arsenic, using well-defined chemical
systems, so that we will also have defined chemical species.  Through
the use of defined chemical systems, we may be able to demonstrate the
mechanisms of action of metallic pollutants on marine and estuarine
shellfish.
REFERENCES

Banner, L.H., C.D. Craft, and D.R. Nimmo.  1975.  "A Saltwater Flow-
  Through Bioassay Method with Controlled Temperature and Salinity,"
  Prog. Fish-Culturist  37;  126-129.

Chester, R.  and J.H. Stoner.  1974.   "The Distribution of Zinc, Nickel,
  Manganese, Cadmium, Copper and Iron in Some Surface Waters from the
  World Ocean," Marine  Chem. 2j 17-32.
                                55

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Eisler, R.  1971.  "Cadmium poisoning in Fundulus heteroclitus^
  (Pisces:  Cyprinodontidae) and other marine organisms," J. Fish.
  Res. Bd. Canada, 28; 1225-1234.

Engel, D.W-, W.G. Sunda, and R.M. Thuotte.  1976.  "The Effects of
  Cupric Ion Activity on the Survival of Eggs and Postlarvae of the
  Spot, Leiostomus xanthurus,"  Annual Report to ERDA, 1976, NMFS
  Beaufort Laboratory, Beaufort, N.C.: 431-436.

Jones, M.B.  1975.  "Synergistic Effects of Salinity, Temperature, and
  Heavy Metals on Mortality and Cosmoregulation in Marine and Estuarine
  Isopods  (Crustacea)," Mar. Biol. ^0: 13-20.

McBrien, D.C.H. and K.A. Hassall.  1965.  "Loss of Cell Potassium by
  Chlorella vulgaris after Contact with Toxic Amounts of Copper Sul-
  fate," Physiol. Plantaram, 18: 1059-1065.

Negilski, D.S.  1976.  "Acute Toxicity of Zinc, Cadmium and Chromium to
  Marine Fishes, Yellow-Eye Mullet (Aldrichetta forsteri C. & V.) and
  Small-Mouthed Hardyhead  (Atherinasoma microstoma Whitley).  Aust. J.
  Mar. Freshwater Res. 27: 137-149.

Sunda, W.G.  1975.  Relationship Between Cupric Ion Activity and the
  Toxicity of Copper to Phytoplankton.  PhD Thesis, Massachusetts
  Institute of Technology, 168 pp.

Sunda, W. and R.R.L. Guillard.  1976. "The Relationship between Cupric
  Ion Activity and the Toxicity of Copper to Phytoplankton."  J. Mar.
  Res. 34; 511-529.

von Westernhagen, H., H. Rosenthal, and K.R. Sperling.  1974.
  "Combined Effects of Cadmium and Salinity on Development and Survival
  of Herring Eggs," Helgolander wiss. Meeresunters. 26; 416-433.

von Westernhagen, H. and V. Dethlefsen.  1975.  "Combined Effects of
  Cadmium and Salinity on Development and Survival of Flounder Eggs,"
  J. Mar. Biol. Ass. U. K. 55; 945-957.

van Winkle, W., Jr.  1968.  "The Effects of Season, Temperature and'
  Salinity on Oxygen Consumption of Bivalve Gill Tissue," Comp. Biochem.
  Physiol. 26:  69-80.
                                56

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         ECOLOGICAL CHARACTERIZATION: AN AID TO DECISION MAKING

                             Howard D. Tait*
     Ecosystem studies by the U.S. Fish and Wildlife Service (FWS) and
funded through the "EPA energy program" are now operational in four
major coastal areas:

1.   Chenier Plain area of southeast Texas and southwest Louisiana

2.   Pacific Northwest

3.   Sea Island - Carolinian

4.   Maine.

In this report, the rationale behind these studies, the progress to
date, and the' work projected for 1977 are reviewed.


RATIONALE FOR ECOLOGICAL CHARACTERIZATION

     Coastal areas, particularly wetlands, estuaries, and flood plains,
are among the most important fish and wildlife areas on the continent.
They are not only important in terms of the birds, mammals, fish, and
shellfish they produce, but also for food, recreation, and refuge for
man.  Rocky coastal areas also serve as vital habitats for certain ma-
rine mammals and seabirds; thus, they command the attention of the FWS.

     Unfortunately, unless policy decisions reflect ecological reality,
the nearshore and onshore areas will receive the brunt of the ecological
impacts of expanding energy programs.  This is true, whether it be outer
continental shelf  (OCS) oil and gas development, tidal power, power
plants, superports for supertankers, or geopressure energy extraction.
The direct impact and the indirect effects must be considered.

     It may be useful to consider some of the decisions and needs for
information associated with the OCS oil and gas program.
*    U.S. Fish and Wildlife Service, Office of Biological Services, Bay
St. Louis, Mississippi 39520.
                                57

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     There are, of course, concerns about the effects of exploratory
and production rigs on marine life.  Unless lease areas are  carefully
selected and operations monitored, there is a potential for  damaging
bottom fauna through discharges of wastes, drill muds, and cuttings.
In addition, there is the obvious hazard of oil spills from  tankers  and
accidents on platforms at sea, and much attention is being given  to
these impacts of the OCS program.  The concern addressed here, however,
is the onshore and nearshore effects of development.

     If there is a substantial find in one of the OCS frontier areas,
there will be a need for support facilities, platform fabrication yards,
pipecoating and marshalling areas, pipelines, and tank forms.  Refin-
eries, gas-processing plants, liquefied natural gas  (LNG) processing
plants, the petrochemical industry, and associated community services
will follow the development.  Most of these facilities require water-
front locations along with dredging and dredge disposal and the filling
of wetlands.  Platform fabrication yards, for example, may occupy
200-1,000 acres of cleared level land adjacent to navigable waterways.
Such a fabrication yard is planned at Cape Charles in Northhampton
County, Virginia.  Environmental impacts are associated with all these
facilities; some can be devastating to fish and wildlife.

     The adverse ecological effects of these developments may include:
turbidity, eutrophication, pollution with toxic materials, basin shoal-
ing and oxygen depletion, benthic habitat degradation, shoreline altera-
tion, saltwater intrusion, and, wetlands loss.  Therefore, it is criti-
cally important that decision makers have available early the essential
information on fish and wildlife requirements, the resources of the
areas, the impacts to be expected, and the options available to mini-
mize environmental problems.

     For instance, the state coastal zone management and fish and game
departments, Federal Power Commission  (FPC), Bureau of Land Management
 (BLM), EPA, and the U.S. Army Corps of Engineers, to name a few, will
want to know where pipelines or pipeline corridors are contemplated.
The FWS would like to be in a position to advise on the optimum routes
from a fish and wildlife habitat viewpoint.  For each alternative route,
we would ask:  where are the nearshore and marsh ecosystems,  and in what
way will they be disturbed by the proposed pipeline route?  Are endan-
gered species or critical habitat present?  Is dredging and spoil dis-
posal involved?  Will nesting birds be disturbed during construction?
Will wetlands be crossed?  Will digging of a pipeline canal allow intru-
sion of saltwater with consequent vegetational changes that could dimin-
ish the food supply, cover, or nesting habitat of some specie.s?  Obvi-
ously, insight is needed about both the components of the ecosystems
involved and how they interact.

     Plainly, decisions regarding OCS development can ultimately have a
large effect on fish and wildlife habitat.  The decision makers need to
know the environmental consequences of alternative courses of action.
                                58

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Recent court and other governmental actions relative  to Lower  Cook  Inlet
and Baltimore Canyon lease areas point up the urgent  need  for  ecological
and other information if further delays in OCS development are to be
avoided.

     It was against this background that the Office of Biological Ser-
vices (OBS) of the FWS decided to initiate an ecological characteriza-
tion program early in 1975 with EPA energy "pass-through"  funds.  It
seemed to us that, through characterizing the coastal areas, FWS would
serve its own interests of protecting habitat and also provide the
many decision makers involved in the OCS and other energy  development
programs with essential ecological information.  The  problems  encoun-
tered with OCS oil and gas development are similar to those associated
with other industrial activity, housing, marinas, bridges, causeways,
bulkheads, and filling of wetlands for highways.  Furthermore, the
characterization of coastal areas would materially assist  state and
local planners and industry in reaching informed decisions about other
developments.
ECOLOGICAL CHARACTERIZATION DEFINED

     The concept of ecological characterization is not very profound or
complicated.  We do consider it a  "common serise" approach to information
gathering and synthesis.

     One feature of ecological characterization that distinguishes it
from conventional*surveys, environmental impact studies, and baseline
studies is its emphasis on the functional and dynamic features of
coastal systems, rather than on the components within the systems.
Baseline studies usually  attempt to establish a basis for detecting
future change in the ecological attributes considered.  An ecological
characterization, on the  other hand, attempts to provide an apprecia-
tion of the existing situation and an understanding ^f ecosystem dy-
namics.

     The OBS is interested in identifying the structure of the biolog-
ical communities and the  values of chemical and physical parameters;
more fundamentally, we are examining the functional relationships among
the various components.   In the characterization, we are attempting to
describe energy flows, nutrient cycles, predator-prey relationships,
and community succession.  We are  also directing particular attention
to environmental variables that affect the number and distribution of
species which are of special interest to man  (e.g., commercial, sport,
and endangered species),  and which are key to the maintenance of eco-
logical stability and productivity.

     The objectives of ecological  characterization, then, are to:

    Develop an understanding of the ecosystem function for a given
     area
                                 59

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    Develop a characterization atlas that presents  the  information on
     ecosystems in graphic illustrations, maps, narrative  sections, and
     descriptive or conceptual models

    Involve potential users in the design and review of the  studies so
     that the studies will be responsive to their needs

    Identify additional needs for information and recommend  needed
     studies.

     The characterization is being undertaken in two phases:   (1) devel-
opment of qualitative information, and  (2) a quantitative phase in
which predictive models are developed and environmental  impact scenar-
ios  run.  To date, the DBS has been heavily engaged  in phase  one:   the
qualitative aspects.
DEVELOPMENT OF QUALITATIVE INFORMATION

     The  following criteria were considered in selecting study areas:
 (1)  schedule of OCS and other probable energy developments affecting the
candidate area;  (2) fish and wildlife values, e.g., presence of National
Wildlife  Refuges, colonially nesting birds, endangered species, winter-
ing  areas;  (3) potential of pertinent information being available;  (4)
size of ecological units in relation to probable cost of doing the
study; and  (5) other ongoing studies.

     We selected the Chenier Plain of southwest Louisiana and southeast
Texas for our first study area because of the wealth of data available
and  because of the area's long experience with oil and gas development.
In addition, it is particularly rich in fish and wildlife values.

     The  Chenier Plain study has been underway now for about 8 months,
and  Energy Resources Corporation (ERGO), the overall contractor, is
responsible for developing and managing the studies, habitat mapping,
and  report preparation  (see Exhibit 1).  Louisiana State University
 (LSU) is  engaged in ecosystem modeling, data assembly, and analyses of
subsystems.  Bendix Applied Science and Technology Division is conduct-
ing  habitat mapping using the MDAS (LANDSAT imagery) system.

     Ecological characterization is intended to serve primarily as a
resource management tool.  Thus, we are involving the management agen-
cies continually during the planning, data collection, initial testing,
and  final evaluation.  In each case, an advisory steering committee has
been established to provide ideas and to assist in locating data.  Indi-
viduals serving on these committees typically are drawn from state fish
and  game, and coastal zone management offices, universities, BLM, EPA,
National Marine Fisheries Service,  and field offices of the service.
                                60

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                  CONCEPTUAL MODEL
    /GENERAL DESCRIPTION Of  \
    / COASMI Win EVITONHENT\


  /  DEFINITION OF BASIC SrSTEHS\

              I
/  IDENTIFICATION OF DRIVING FORCES \

TION OF
NG COHCCNUAl
s


VADlnCILITr IN
WIVINT. FORCES
           SELECT ion OF rimcTlONim tcoior.icfli
TSIS OF KET
CTUWL
OBENTS

rtmcTiOHAL
COASTAL SYS
(FINALIZED

OF
TEHS
MODEL)

AHALTSIS OF
FUIICTIONAL
PROCESSES
KIT1IIH AND
BETWEEN UBITS



ANALYSIS OF
STRESS FACTOI

OUTLINE FOR DATA

!

             EVALUATION OF USER HEED
             AND POTENTIAL DATA SOURCES
                                                                            OATA COLLECTION


1 SEARCH OF PUBLISHED I UNPUBLISHED
DATA. ONGOING RESEARCH
             DESIGHATION OF NAP RESOLUTION
                                                               HIBI- CHABACTIRIZBTIOJ|[	[ ATLAS PREPAKAIIOH |
                                                                             DATA SOURCE
                                                                             APPENOII
                                                                             PMPABAriOH
                                                                                                              FINAL REPORT I
                                                                                                              PREPARATION
EXHIBIT  1:    APPROACH  TO CHARACTERIZING  THE  STUDY  AREA

-------
     In general, each study involves the following tasks:

    Definition of the boundaries of the study area along with an ini-
     tial canvass of available data

    Literature search

    Conceptual model development

    Definition of user needs, usually through questionnaires and inter-
     views

    Designation of map scales and resolution

    Data collection

    Characterization of a pilot area

    Evaluation and revision of methodology based on user evaluation of
     pilot study

    Preparation of characterization atlas of entire area

    Preparation of data source appendix

    Summary of project outputs.

Exhibit 2 presents the status of these studies.   When these tasks are
completed, the FWS will conduct a series of user-oriented seminars to
help ensure that the utility of the products is  understood and that the
availability of the reports is known.
                               62

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                   EXHIBIT 2:   STATUS OF  STUDIES
 Coastal Area
                  Date Started
                               Contractor
 Chenier Plain
                  4/5/76
 Sea Island
 Maine
                  2/23/77
                  2/23/77
 Pacific Northwest
                  2/7/77
Energy Resources Co., Inc.
Cambridge, Massachusetts

Center for Wetland Resources
Louisiana State University
Baton Rouge,  Louisiana

Bendix Applied Science and Technology
Ann Arbor, Michigan

South Carolina Wildlife and Marine Resources
  Department
Marine Resources Division
Charleston, South Carolina

Energy Resources Co., Inc.
Cambridge, Massachusetts

Bigelow Laboratory
McKown Point, Maine

Bendix Applied Science and Technology
Ann Arbor, Michigan

Ryctanan/Edgerley/Tomlinson and Associates, Inc.
(Division of  Envirodyne Engineers, Inc.)
St. Louis, Missouri
FUTURE  PROPOSED  STUDIES

     The studies currently  underway will identify gaps in  information
needed  to more adequately understand  the ecosystems and to provide the
data needed by management.   On a case-by-case basis, we will evaluate
the practicality of conducting Phase  two.  This  phase involves mathemat-
ical modeling to attempt prediction of specific  impacts, given certain-
scenarios.  We will also improve the  data base with a view to developing
digitized, geobased inputs  for computer modeling and mapping.

     Numerous coastal areas have been proposed for ecological character-
ization.  We are conducting preliminary planning for the following
areas:

    Yukon Delta, a vitally important waterfowl-producing  area of Alaska

    Lake St. Clair, a water route for fuel transport and  an important
     waterfowl and fishery  area

    Eastern Gulf of Mexico (Louisiana, Mississippi, Florida), a site  of
     potential geopressure  development, with heavy tanker  traffic and
     superport development

    Southern California, expanding DCS development
                                   63

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    Mid-Atlantic, preferred DCS area for resource potential and heavy
     tanker traffic

    Pacific Islands (Hawaii, American Samoa and Trust Territories).

     A recently completed summary of state information needs prepared
jointly by the Office of Coastal Zone Management, National Oceanographic
and Atmospheric Administration, and the BLM confirms our belief that
ecological characterization studies should be continued and expedited
if possible.  The following is quoted from page 12 of the January 1977
report.

     "Highest Priority"

     The greatest needs for information relate to the potential
     effect of oil spills on the nearshore environment,  the loca-
     tion and impacts of pipelines and onshore facilities,  OCS-
     related employment and the fiscal impacts upon local commu-
     nities. "
                               64

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AN ECOLOGICAL CHARACTERIZATION OF THE CHENIER PLAIN OF SOUTHWESTERN
   LOUISIANA AND SOUTHEASTERN TEXAS: OBJECTIVES AND METHODOLOGIES

                            Janie Harris*
      As the nation continu.es to move toward increased energy develop-
 ment and industrial expansion, we are relying more and more heavily on
 the environmental impact statement (EIS)  process to guide us in our
 efforts to minimize adverse ecological impacts.  However, the current
 method of incorporating biological and ecological information into
 EISs has not proven to be a satisfactory predictive tool.  Existing
 ecological impact assessment methodologies have not adequately addressed
 the complex chain of biological events associated with human activities.
 The coastal ecosystems team of the Office of Biological Services of the
 U.S. Fish and Wildlife Service has recently designed a new approach to
 the investigation of ecological systems.   The purpose of this program is
 to provide a more systematic procedure for both the examination of
 biological systems and the prediction -of impacts on these systems.  This
 new approach, termed "ecological characterization/' is being applied
 first to the analysis of coastal systems.

      Energy Resources Company of Cambridge, Massachusetts, together
 with their subcontractors, the Louisiana State University, Center for
 Wetland Resources, and Bendix Applied Science and Technology, Ann Arbor,
 Michigan, have been selected to conduct the first of these coastal eco-
 logical characterizations in the Chenier Plain of southwest Louisiana
 and southeast Texas.

      The distinguishing feature of the characterization, which makes it
 unique in comparison to conventional biological studies, inventories,
 and other environmental programs, is its emphasis on the functional and
 dynamic features of coastal systems.  By studying these functional
 attributes of the system, we hope to come closer not only to understand-
 ing the driving forces of the represented ecosystems, but also to under-
 standing how the system as a whole is affected by various resource
 development and environmental modifications.
 *    Energy Resources Company, Inc., 185 Alewife Brook Parkway,
 Cambridge, Massachusetts  02138.
                                 65

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PROGRAM OBJECTIVES

     The objectives of the Chenier Plain program as outlined by the
U.S. Fish and Wildlife Service are essentially fourfold.  The first
objective is to develop an understanding of the interactive mechanisms
and driving forces that govern the area's ecological dynamics.  That  is,
we are interested not only in identifying the structure of the biolog-
ical communities and the values of various associated physical and
chemical parameters but, more fundamentally, we are examining the func-
tional relationships among the various biological and nonbiological
system components.  These "functional aspects" include energy and mate-
rial flows, nutrient cycles, predator/prey relationships, species com-
petition, breeding requirements, community succession, and the like.
In connection with this effort, we are directing particular attention
to environmental variables that affect the number and distribution of
the indigenous species which are of special interest to man (e.g.,
commercial, sport, and endangered species) and those which are key to
the maintenance of ecological stability and productivity.

     The second objective is to develop a characterization atlas that
presents these structural and dynamic features clearly in graphic
illustrations, maps, reports, and descriptive models for use by local
resource managers and environmental planners.

     The third objective is to specify additional information that may
be required to characterize the Chenier Plain more completely and to
recommend special field studies to generate these data.

     The fourth objective is to design an effective and expeditious
ecological characterization methodology for future studies, based on our
experience gained during the Chenier Plain program.

     However, the achievement of these objectives through the perfor-
mance of the characterization program is not enough, in itself,  to
change the current methods of ecological impact prediction.  Implicit
in the development of a characterization program is the need to work
closely with the local resource management community and to enlist their
suggestions and technical support.  Without the enthusiastic support of
this community, the characterization program products are unlikely to
be used in planning, impact prediction, and resource management efforts.
Thus, we are maintaining close communication with the local experts and
managers and are soliciting their guidance, criticism, and suggestions
so that the final products will, in fact, be responsive to their needs.

     This discussion of the Chenier Plain program will address three
major topics:

1.   Brief overview of the Chenier Plain coastal system

2.   Presentation of our basic methodological approach to the
     characterization effort
                               66

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     3.  Description  of the outputs that we are producing and an expla-
         nation of  their utility to investigators and  resource managers.

CHENIER PLAIN GENERAL DESCRIPTION

     The Chenier  Plain, as defined for the purposes  of our study, is
that coastal area westward of Vermilion Bay, Louisiana,  and eastward
of Calveston Bay, Texas.  It is bordered on the north  roughly by the
5 ft inland elevation line and on the south by the Gulf  of Mexico
30 ft depth contour (see Exhibit 1).
          DOTTED LIME INDICATES APPROXIMATE
          BOUNDARY OF CHENIER PLAIN STUDY AREA
               EXHIBIT 1:   CHENIER PLAIN STUDY AREA
      This area is predominantly a marsh and open water system, demon-
 strating virtually no  relief features aside from the characteristic
 Chenier ridges, which  run roughly parallel to the shoreline.  The
 ridges, which run from a  few inches to more than 10 ft in elevation,
                                 67

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 are approximately 100-1500 ft wide and may extend along the coastline
 for distances of up to 30 miles.

      The presence of these cheniers, which decrease in size and number
 as one moves westward through the study area, raises some interesting
 questions relative to the plain's geological history.   The Chenier Plain
 exists as a result of natural accretion of Mississippi River sediments.
 However, unlike the Deltaic Plain, which grew from the direct deposi-
 tion of the Mississippi's sediments, the depositional  processes contri-
 buting to the formation of the Chenier Plain involved  sediment trans-
 ported westward by prevailing longshore currents.   These sediments were
 then deposited along the shoreline as mud flats.   These accretionary
 mud flat areas were then colonized by emergent vegetation, which sta-
 bilized the flats and contributed to the continual shoreline prograda-
 tion and the establishment of a vast marsh system.

      Numerous times during the area's geologic history, the Mississippi
 River shifted its depositional centers.  During westward discharges, the
 plain continued to prograde as described above.  During easterly shifts,
 however, the sediment supply was decreased and accretion slowed or
 stopped completely, resulting in coastal retreat,  as subsidence exceeded
 sedimentation.  This shoreline retreat was coupled with the sorting of
 shell fragments and sands fr'om the finer clays- and silts by wave action.
 This process resulted' in the formation of beaches  overlying the mud flat
 and marsh areas.   As the sediment source was renewed by a subsequent
 shift back to a westerly Mississippi River discharge pattern,  the mud
,- flats and marshes would again prograde seaward of  the  beaches,  which
 then became stranded inland as Chenier Ridges.

      The cheniers are very important to the determination of this area's
 ecological processes, hydrological regime, and human population distri-
 butions.  They provide pastureland and access routes into the marsh
 areas for terrestrial species.  They are suitable  for  colonization by
 large trees, particularly oaks, thus providing a  habitat for various
 bird species.

      The Chenier  Plain area is characterized primarily by various types
 of marsh and aquatic ecosystems.   These wetland areas  are rich in renew-
 able and nonrenewable resources and are subjected  to a wide variety of
 land use practices, most of which involve water level  regulation.   The
 major land uses,  which direct these management schemes, include oil and
 gas production; rice, soybean, and crawfish farming; fish and shellfish
 harvesting; fur trapping; and alligator and waterfowl  hunting;  as well
 as maintenance of wildlife refuges and waterfowl preserves.   Thus, the
 employed management strategies are designed to maximize the productivity
 of the Chenier Plain with respect to these resources.
                                 68

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METHODOLOGY

     The first phase of the characterization program  involves a qualita-
tive assessment of the major functional  interactions  within  the Chenier
Plain system.  In the second phase, we plan to employ a variety of ana-
lytical techniques to quantify the ecological relationships  and energy
and material flows.  The second phase, then, will allow us to develop
predictive models and to run environmental impact scenarios.  Because
the second phase has not yet been initiated, this discussion will be
restricted to the methodology and products of the first phase, which
involves six separate tasks:

1.   Development of a conceptual model and the determination of data
     needs

2.   Data collection/data  synthesis

3.   Production of a working habitat map

4.   Pilot characterization and methodology evaluation

5.   Final characterization

6.   Production of characterization atlas and reports.

     The conceptual model, which is the  guiding element of the character-
ization, is essentially a  portrayal of the principal  ecosystem compo-
nents and fundamental relationships that link them.   By identifying the
major features and interactions, this model provides  the focus for the
requisite analysis as well as the framework for data  collection.

     The first step in conceptual model  development was to divide the
Chenier Plain into a set of subsystems,  each of which is internally
homogeneous with respect to certain key  variables.  This subdivision was
performed on three levels  of complexity  as based on six key  environmen-
tal variables:  water flow patterns, elevation, vegetation type, inunda-
tion and salinity regime,  and man-imposed environmental modifications.

     The three levels of complexity are:  the regional level, the drain-
age basin level, and the habitat level.  At the broadest level, the
model treats the geologic  history and geomorphological features of the
entire area.  At the intermediate level, the drainage basin, the model
identifies six distinct hydrologic units and addresses the ecological
interactions among the various habitat types within each basin.  We
believe that the interhabitat interactions defined at this level are
largely governed by the water flow patterns, inundation characteristics,
and salinity regime.  The  habitat level  of the conceptual model ad-
dresses the major community assemblages  that characterize the plain and
provides the framework for defining and  investigating the functional re-
lationships among the physical, chemical, biological, and ecological com-
ponents within the 14 major habitats identified in the Chenier Plain.


                                69

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     These 14 habitats are identified as:

     Aquatic Habitats
     Wetland Habitats
     Upland Habitats
 1.  Inland open water systems,
     including both lakes and bayous

 2.  Nearshore gulf waters

 2a. Mudflats

 3.  Salt marshes (including small
     ponds)

 4.  Brackish marshes (including small
     ponds)

 5.  Intermediate marshes (including
     small ponds)

 6.  Fresh marshes (including small
     ponds)

 7.  Swamp forests

 8.  Upland forest (including pine
     forest)

 9.  Ridges (cheniers, artificial and
     natural levees, and spoil banks)

10.  Beaches

11.  Rice fields and other managed
     crop areas

12.  Pasture lands and grassland prairies

13.  Managed marshes (for waterfowl,
     etc.)

14.  Urban areas and industrial sites.
     For each of the 14 components of the habitat level of the model,
a schematic diagram was developed to present the major components and
the  interactions occurring among them.  This schematic is supplemented
by an interaction matrix that specifies the nature, direction, and
variability of the important interactions.

     Once the key features and interactions were specified for each of
these abstracted habitats types, we mapped the distribution of the
habitats as they occur in the Chenier Plain.  This mapping allowed us to
aggregate the available data sets by habitat type and to investigate and
     Altered Habitats
                                 70

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compare the functional interactions  that  characterize  similar  and
dissimilar systems.

     Our procedure for generating  this working  habitat map  involved  the
processing of LANDSAT satellite  imagery of  the  area.   The LANDSAT  system
distinguishes different habitat  types based on  their reflective prop-
erties.  A specific area of the  Chenier Plain was  identified as a
particular habitat type by a ground  truth expert.   The automated LANDSAT
system then identified all areas that have  the  same reflective proper-
ties as the sample area and assigned them a particular color code.   The
habitat map was then refined by  adding information from aerial photo-
graphs, existing maps, literature, and ground truth to produce a suf-
ficiently detailed map.

     The next step in the program  is the  performance of a pilot char-
acterization for a portion of  the  total study area.  This task enables
us to test our conceptual model  and  characterization methodology on  a
small area and to generate a pilot study  atlas  to  review selected  dis-
play techniques.  Thus, we can evaluate our approach and refine it
before the characterization of the entire area  is  undertaken.
PROJECT OUTPUTS

     At the conclusion  of  the  first phase of  the Chenier program, we
will have six major outputs  that will be available  through the U.S.
Fish and Wildlife  Service.   These products  include:  the finalized
conceptual model document, the characterization atlas, a bibliography
of relevant information sources, a computerized data base, a report
recommending-field studies that will help fill crucial data gaps, and
a s^t of methodological recommendations to  guide future characteriza-
tion efforts.

     Clearly, the  characterization atlas, the bibliography, and the
computerized data  base  will  most directly address the information needs
of investigators,  resource managers, and people responsible for the
preparation of EISs.  The  conceptual model, field study recommendations,
and methodology outline are  being prepared  primarily for use by
investigators involved  in  future characterization studies and those
who update the ongoing  studies.

     Hopefully, the characterization approach to the examination of
ecological systems, by  focusing on the dynamic and  functional aspects
of the system, will help lead  scientists away from  the temptation to
describe ecosystems as  if  they were static  in time  and space.  Our
ultimate objective, then,  is to present the Chenier Plain coastal
system as a set of functionally interactive subsystems, defined in such
a way that impacts on important biological  communities can be pre-
dicted and evaluated in the  context of the  basic interacting forces
which define the system in toto.  Moreover, many of the elements of the
characterization effort are  important not only because they allow us to
                                 71

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understand the primary and secondary environmental modifications
expected to result from a proposed resource development but also because
they bring us closer to the understanding of the composite system and
how it functions.
                               72

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DISCUSSION

     Dr. Lefcort;  The Fish and Wildlife  Service  is  supporting  a  con-
tract with the Conservation Foundation to assess  the onshore  impact of
offshore development.  In this computerized  data  retrieval  system, you
query the computer and you get the answer in terms of what  is available,
everything you have always wanted to know.

     In terms of OCS development, these two  projects are quite  similar.
Where are the areas of overlap?

     Dr. Tait;  They are not overlapping.  They are very complementary.
The computerized aspect of the Conservation  Foundation Project  is still
in an experimental stage.  It has a lot of promise because  there  is a
lot of data that can be fed into that system:  the quantitative data
from these characterizations; information on endangered species and
their habitats; and the material that the Conservation Foundation itself
will generate relative to onshore impacts.

     Dr. Palmisano;  The Conservation Foundation  contract is  kind of a
cookbook of how to put together all of the complex activities associ-
ated with OCS development.  It attempts to present a typical  sequence
of events related to OCS, beginning with  the leasing all the  way through
to development, and put this  information  in  a time and space  perspec-
tive.  Associated with that is an analysis of the impacts of  these
activities on resources.  This model can  be  applied anywhere  in the
United States, on any coast, under any situation.  However, you have to
go to a specific area, such as the Chenier Plains, and look at  the biol-
ogy and ecology of that particular area,  and apply the guidance in the
Conservation Foundation Guidebook.

     It first outlines a broad program, OCS,  say, for mid-Atlantic.  It
then describes the project, which is a broad activity that probably will
require an impact statement such as the location  of a nonshore  facility
storage tank facility.  This project will have a  subproject,  such as a
navigation channel and a subproject has an activity.  An activity is
the level at which permits are usually issued, for dredging,  bulkhead-
ing, land filling, and so forth.  In this way you can look  across an
entire OCS development program.

     Dr. Barrientos;  Will you get storm  search data?

     Dr. Tait:  I think any storm search  data that we have  will be that
immediately available from other records.
                                 73

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                THE CONSTRUCTION OF A CONCEPTUAL MODEL
      OF THE CHENIER PLAIN COASTAL ECOSYSTEM IN TEXAS AND LOUISIANA
             L.M. Bahr, Jr.,* J.W. Day, Jr.,* T. Gayle,t J.G. Gosselink,*
                  C.S. Hopkinson,* D. Stellar,* and J. Harris**
     Increasing interest in coastal areas on  the part  of  environmen-
talists, developers, and managers has generated the  need  to  understand
the function of productive and fragile areas  and to  predict  the  effects
of further alterations to them.  The term "function,"  as  used  through-
out the following description of the Chenier  Plain conceptual  model,  is
intended to describe the mechanics of the ecosystem, i.e., the pathways
and processes by which energy and matter is captured,  transferred,
partitioned, stored, cycled, and degraded by  the system.  Examples of
functional processes include primary production, water flow, trophic
exchanges, and animal migrations.  A functional understanding  of an
ecosystem includes much more than an inventory of important  physical
parameters and organisms; it requires a holistic, systems-level  analysis
to identify all important interactions among  biological and  physical
components -of the system and all important control features  and  feedback
mechanisms.

     The U.S. Fish and Wildlife Service  (Department  of the Interior)
decided in late 1975 to fund an exhaustive study of  the Chenier  Plain
coastal ecosystem(s) of Southeastern Texas and Southwestern  Louisiana
(Galveston Bay, Texas, to Vermilion Bay, Louisiana), in which  the area
would be characterized ecologically by the development of a  conceptual
model of the system and a synthesis of all extant data.   This  character-
ization was designed to serve as a pilot study for future projects that
will eventually describe all U.S. coastal ecosystems.   The specific
request was for a "description of the important resources and  processes
comprising the ecosystem and an understanding of their functional
relationships."  The first requirement of this study (and the  key to
 *  Center  for Wetland  Resources,  Louisiana State  University,  Baton
 Rouge, Louisiana'  70803.

 t  Center  for Wetlands, University  of  Florida,  Gainesville, Florida
 32601

 ** Energy  Resources Company,  185  Alewife  Brook  Parkway,  Cambridge,
 Massachusetts  02138.
                                 75

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the entire project) was the formulation of a conceptual model of the
ecosystem(s).   The model was to consist of a schematic framework of
ecosystem function in which all important processes and interactions
among components would be identified in a qualitative manner.  The
completed model would identify data requirements and gaps, and set the
stage for the two remaining portions of the study, a characterization
atlas, and a quantitative ecological simulation model of the study area
that could be used to aid in making management decisions.
DESCRIPTION OF THE PROBLEM

     Any ecological model of the Louisiana-Texas Chenier Plain must take
into account at least the following four factors:

1.   Spatial heterogeneity.  The area described as the Chenier Plain
     (see Exhibit 1) is highly variable in space; from east to west, it
     is broken up by a series of rivers flowing southward into the Gulf,
     through lakes of different sizes and salinities, and over thousands
     of square miles of wetland.  The wetlands themselves are not at all
     homogeneous; vegetation ranges from pure stands of saline oyster
     grass  (Spartina alterniflora) to fresh water bull tongue (Sagit-
     taria falcata) and maidencane (Panicum hemitomum).  They are cut
     by elevated cheniers or ridges, which function ecologically in a
     very different manner than the adjacent wetlands.  Large areas,
     both wetlands and highlands, have been modified for agriculture or
    - are managed for waterfowl or furbearers.  The region is far from
     homogeneous, and any model that ignores this will produce informa-
     tion of limited management value.

2.   Ecological or functional complexity.  Aside from this spatial
     heterogeneity, the ecological food web is enormously complex within
     any small, fairly homogeneous area.  On the whole, it is poorly
     understood.

3.   Time scale of events.  Events of ecological interest in the Chenier
     Plain occur at the scale of hours, days, or seasons for many
     biological processes; years or tens of years for many cumulative
     human impacts, such as canal dredging and eutrophication; and
     thousands of years for geological processes that determine the
     physiography of the whole region.  It is difficult to visualize a
     useful model that can simultaneously simulate geological processes
     and microbial kinetics in terms that are useful to a manager.

4.   Management needs.  In addition to the above considerations, the
     model must enable a manager to evaluate the consequences of alter-
     nate management strategies at appropriate levels of spatial,
     ecological, and temporal resolution.  Existing models cover a wide
     range of approaches, including strategies to exploit or manage
     single commercial species, such as fishery models (Wagner, 1969) or
                                 76

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  30'00'
    S'00'
                                                               -50" __-
                                                                       50	IpOKm
                                                                    90'00'
EXHIBIT 1:   THE CHENIER PLAIN  REGION OF SOUTHWESTERN LOUISIANA AND SOUTHEASTERN TEXAS

-------
alligator models  (Nichols et al.,  1976); models that treat eco-
systems as homogeneous in space in order to elaborate the energetic
interactions  (Patten et a^., 1975; Wiegert et. al_., 1975); or models
that treat spatial  heterogeneity but consider only a limited number
of chemical or  biological parameters (Kremer and Nixon, 1975) and
dramatically  simplified,  dynamic world view models (Forrester, 1971) .
THE STRUCTURE  OF  THE CONCEPTUAL MODEL

      In  its present form,  the conceptual model is a hierarchy of nested
models that divide  the  Plain into three levels of spatial, functional,
and temporal  resolution or the habitat, the basin, and the region  (see
Exhibit  2).  By focusing attention at the appropriate level, this hier-
archy allows the  characterization and analysis of the Chenier Plain and
the handling of the four issues of spatial heterogeneity, functional
complexity, time  scale  problems, and management needs.
       Region
Chenier Plain
                                                         7000
                                                              Years
        Basin


Calcasieu
Basin



                                                         T-700 /ears
       Habitat
                                                         .07 - 70 /ears
                       Open Water
                                       Salt Marsh
         EXHIBIT 2:   SCHEMATIC ILLUSTRATING THE TIME SCALE
           OF EVENTS OF INTEREST FROM HABITAT TO REGION
     The smallest identifiable  ecological  units  are called habitats.
Wherever a particular habitat occurs  on  the  Chenier Plain, it is recog-
nizable as the same basic  functional  unit, and can, therefore, be
                                 78

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treated as homogeneous, even though we recognize  the  existence of  grad-
ients, specialized niches, and discontinuities  within a  habitat.*

     Each habitat is a complex ecological  system.  Each  is  character-
ized by its own species, carrying capacities  for  those species,  levels
of production, food web, nutrient cycles,  and physical inputs, wherever
it is found.  The time scale of important  events  is often seasonal, and
short-term impacts are important at this level.   Most habitats are
intuitively distinct.  For example, aquatic areas are quite different
from upland forests; however, different kinds of  natural wetlands  are
not so clearly unique.

     For the Chenier Plain, we have identified  and mapped 10 natural
habitats:  nearshore Gulf; inland open water; salt, brackish, inter-
mediate, and fresh marsh; wetland forest;  upland  forest; beaches;  and
cheniers and ridges.  Large areas have been modified  by human activity,
which we have catalogued into four additional habitats as impounded
marshes, pastures, rice and crop habitat,  and urban habitat.

     Habitats interact at a broader level  of  resolution, i.e., the
basin.  Drainage basins represent perhaps  the most natural ecological
units of the region, because each basin is integrated by the flow of
water over and through it.  Six fairly distinct basins have been identi-
fied on the Chenier  Plain  (see Exhibit 3,  and Bahr _et _al. , 1977).  Each
basin has its own hydrodynamic characteristics  determined by upstream
flow, elevation and  slope of the basin, and extent of its connection
with the Gulf via tidal passes.

     At the basin level, emergent properties  such as  interhabitat inter-
actions become important, and intrahabitat details are ignored by
treating each habitat as a black box with  known inputs and outputs.  The
system is no longer  treated as homogeneous in space.  Most significant
changes occur through large-scale or cumulative impacts over a period of
time measured in years, not in days or seasons.   Examples include:
impacts of deep shipping channels on saltwater  intrusion; gradual
changes in hydrology associated with stream channelization; continuous
canal dredging and associated spoil bank formation; and cumulative wet-
land drainage, a little at a time, for urban  and  industrial development.

     The six basins  of the Chenier Plain are  unified  by a common
geologic history.  Geologic processes unify the region and determine the
specific characteristics  (e.g., slope, relief)  of each basin, but for
most purposes these  characteristics can be considered as unchanging in
the time scale of basin or habitat-level phenomena.
     See Bahr  et al.  (1977),  for  justification of habitat concept.
                                 79

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00
o
                                EXHIBIT 3:  WATER IS THE PRIMARY INTEGRATING ELEMENT

                                  IN THE SIX BASINS IN THE CHENIER PLAIN REGION

-------
     We have developed conceptual models at  all  three  levels  of  the
hierarchy, but for the reasons discussed below the  emphasis of the
quantitative model is at the basin  level.

     The regional model is primarily a model of  geological processes
(see Exhibit 4), which are not strongly influenced  by  man except as he
controls the flow of the Mississippi River.  Beach  and mudflat growth
or erosion depends on the interaction of depositional  and erosional
processes that use and rework longshore and beach sediments.  When
deposition predominates, mudflats build offshore of the beach.   During
periods of erosion these mudlfats are reworked and  winnowed to leave
sand beach ridges.  When net deposition occurs again and mudflats build
out, the beaches are stranded in inland cheniers.

     To give a qualitative functional understanding of the habitat and
guide the acquisition of data, complex habitat level models have been
drawn for each of 14 habitats.  However, quantitative  mathematical
models at this level are more a subjective art than an objective
science  (see Exhibits 5-7).

     For  several practical reasons, we have  concluded  that major
emphasis  for management purposes  should be placed at the basin level.
Development of a  functional mathematical model to simulate the full
range of  ecological  complexity of  the Chenier Plain would represent a
staggering undertaking.  Furthermore, when we consider the success of
similar models .by others and  our  understanding of basic ecological
interactions,  such a model would  not necessarily be very useful.  In
addition, both the major kinds of  manageable processes and the time
scales of manageable events appear to occur  at the  basin level;  changes
in area of habitats, such  as  the  gradual increase in agricultural area
at the expense of wetland; eutrophication from urban growth;  and, per-
haps most significant,  the gradual cumulative changes  in hydrology
associated with dredging many small canals over  a period of many years.

     An additional consideration  is our ignorance of the internal work-
ings of a habitat.   Thus,  we  manipulate habitats at some peril to the
function  of the whole  system.  A  fcetter approach to management is to
recognize that  certain  renewable  resources or nonresources  (Ehrenfeld,
1976) are associated with  each habitat and that  to  protect the resource
one must  protect  the habitat.
 ASSUMPTIONS OF THE BASIN LEVEL MODEL

      The basin level model was developed with the following assumptions:

 1.    A habitat can be treated as a "black box."   That is,  a habitat
      normally characterized by certain plant species  with  a predictable
      level  of primary production.   Given this level of primary produc-
      tion,  the habitat will have a "natural resource  potential" (NRP),
      which  is quantifiable as a yield or carrying capacity for certain


                                 81

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CD
to
                                        Stranded
                                         Chenier
                                          Plain
                                         Marsh
                                                                                                        Erosion
Erosion
                            EXHIBIT  4:   DIAGRAMMATIC  MODEL OF REGION-LEVEL  GEOLOGICAL PROCESSES

-------
EXHIBIT 5:  DIAGRAMMATIC REPRESENTATION OF AQUATIC HABITATS
  [The habitat is represented as a water storage module  (1),
  with water flows controlled by upstream and tidal head  (2).
  Water carries nutrients, salts, toxins, and sediments into
  and out of the habitat.  These materials control the species
  present and productivity of phytoplankton  (3), floating
  aquatics  (4), and benthic producers  (5), which in turn
  support consumer groups  (6, 7, 8, 9).  Organic detritus  (10)
  is an important organic source to the system.]
                             83

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   UPSTREAM HIVED
       1
    DOWNSTREAM t Sun
    TIDAL WATER >v
EXHIBIT 6:  DIAGRAMMATIC REPRESENTATION  OF WETLAND HABITATS
  [The habitat is shown as a producer  (1)  that exports organic
  carbon.  Upstream  (2) and downstream (3)  water flows control
  the hydrologic regime of the wetland (4), and the flux of
  organic carbon, nutrients, toxins  and  sediments.  Emergent
  producers (5) and producers in  small ponds (6) feed detritus
  to consumers (7, 8, 9), which migrate  through wetlands, are
  hunted and trapped.  Habitat area  (10)  is controlled by
  natural processes  (11), and by  urban and agricultural
  development  (12).]
                             84

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                Toxic
                Stress
                                                              Toxic
                                                              Stress
EXHIBIT 7:  DIAGRAMMATIC REPRESENTATION OF MANAGED HABITATS,
  PASTURE  (1), RICE  (2),  AND MANAGED MARSH (3).  [Much of this
  area is maintained  by  burning,  cultivation, draining and/or
  controlled flooding, at the expense of wetlands.   Food chains
  are simplified, but considerable interaction occurs with adjacent
  habitats  (4, 5).]
                               85

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    species of interest to man (see Exhibit 8).  For instance, a fresh
    marsh will have a high potential for nutria (^90 pelts/100 acres)
    and dabbling ducks  (peak density, 270 ducks/100 acres) but a rather
    poor potential for muskrats,  which are concentrated in the brackish
    marsh  (^40 pelts/100 acres).
             PLANT                        NATURAL RESOURCE
       PHOTOSYNTHESIS                        POTENTIAL
           mt/h_/w	. FRESH MARSH \	  90 Nutria/100 Acres
           15mt/ha/yr	=H    HABITAT     /^ 270 Ducks/100 Acres
                                              NURSERY FUNCTION:
                                              15kg Shrimp/Acre
           EXHIBIT 8:  MATERIAL RESOURCE POTENTIAL OF A HABITAT
             IS DETERMINED BY LEVEL OF PLANT PRODUCTION
     This concept is  implied  in  the notion of "carrying capacity," which
     is extensively used  in wildlife management.  Identification of the
     NRP for a particular habitat does not demand a detailed under-
     standing of the  processes that result in carrying capacities for
     specific organisms.   Rather, it is an empirical observation of the
     value of a certain kind  of  habitat for certain species.

2.    Natural habitats are determined by a set of physical environ-
     mental parameters.   It is generally felt that ecological
     succession in wetland habitats is controlled by the physical
     environment, i.e., allogenic processes dominate over autogenic
     ones (Odum, 1971).   Thus, if the driving  (controlling) environ-
     mental forces (such  as hydrologic regime) change, the  habitat will
     change its identity  and  become a different habitat.  An example of
     this process is  the  gradual intrusion of salinity in the Barataria
     Basin of Louisiana,  which is causing a landward shift  in the
     boundary between salt and brackish marsh and increasing the area
     of salt marsh habitat at the expense of brackish marsh.  For wet-
     lands,  the major controlling parameters are salinity and the
     inundation regime (see Exhibit 9).

3.    Water is the integrating mechanism of the basin  (see Exhibit  10).
     Many of the interactions among habitats, such as organic and
     nutrient fluxes, are dependent on water flows.  Aquatic consumers
     move among aquatic and wetland habitats via flooding waters.   In
                                86

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 80
  60

* 50
tW

|40
30

  20

  10
Brackish Marsh
                 5             10
                          Salinity %o
                        Habitat "niches"
                                             15
                                  from Chabreclc, 1972


   EXHIBIT 9:  WHEN WETLAND HABITATS ARE PLOTTED ON A GRAPH
      OF SALINITY  OF FLOODING WATERS VS SOIL ORGANIC CONCEN-
      TRATION, THE "SPACE" OCCUPIED (MEAN   SD OF EACH PARAMETER)
      BY SALT MARSH OVERLAPS BRACKISH MARSH  VERY LITTLE
                            87

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00
CO
                  EXHIBIT 10:   A DIAGRAMMATIC  REPRESENTATION OF  A COASTAL BASIN [The basin is
                    represented as  a water  storage module,  in which water (and dissolved and
                    suspended  materials)  flow  is  controlled by the upstream and downstream water
                    regime.  Habitat area,  quality,  and  interaction are  controlled by these flows.]

-------
     addition, changes in the pattern  of  inundation (i.e.,  frequency,
     depth, and duration of flooding)  mediate changes  from one habitat
     to another.

     A major identifiable characteristic  of a drainage basin is the
change in area of different habitats,  constrained by the constant size
of the whole basin.  Historical  rates  of  change of habitats within a
basin can be determined with accuracy  from old maps (see Exhibit 11).
The change in area of habitat within a basin is controlled by natural
and man-related processes.  Major  shifts  are from natural wetland to
open water, impounded wetlands,  agriculture, and to other developments.
                                 OTHER AREA
                                RIDGES. FOREST
                                 RECLAIMED!
                                  WETLAND I
                                 DEVELOPED
            EXHIBIT  11:   SCHEMATIC ILLUSTRATING THE  CHANGE
               IN AREA OF  HABITAT WITHIN A BASIN


     For management  purposes,  we can project changes in habitat area
from our knowledge of hydrologic processes of the basin and of the ways
in which modification of  these processes changes habitats.   In practice,
a good two-dimensional  hydrodynamic model is necessary to predict
hydrologic changes,  salinity intrusion, etc., resulting from canal
dredging and  spoil deposition.*
 *  The Louisiana State university Center for Wetland Resources has such
 a model operating for Barataria Bay, Louisiana, but assessment of
 hydrologic  changes in the Chenier Plain study is still primarily quali-
 tative .
                                  89

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     In addition to changes in habitat area, the productivity of a unit
area (the resource potential per unit area) can be changed by such pro-
cesses as eutrophication, introduction of pesticides and other toxins,
water level control by weirs and levees, and overharvesting of existing
resources (see Exhibit 12).
                                                  XJRBAN AND
                                                  INDUSTRIAL;
                                                  DEVELOPMENT
                                                          HARVEST
            EXHIBIT  12:  SCHEMATIC ILLUSTRATING THE CHANGE  IN
              THE  TOTAL NATURAL RESOURCE POTENTIAL OF A BASIN
      From  a  knowledge  of  the  current natural resource potential  of each
 habitat  in a ba.sin  and of areal  and qualitative  changes  in  each  habitat
 over  time  (or because  of  human intervention), one  can project the  total
 change in  the basin NRP-   This change  (which is  almost always negative)
 can then be  compared with the expected benefits  of the activity  to be
 managed.

      As  an example,  the Calcasieu basin  in  the Chenier Plain is  esti-
 mated to support  total primary plant production  (a rough index of  the
 total value  of  the  natural basin landscape) of 2.3 million  metric  tons
 per year.  This produces  an estimated 1,000 mt of  shrimp, 7,700  mt
 menhaden,  5  million sport fishing efforts,  222,000 dabbling ducks  at
 peak  density, and 49,000  fur  pelts.

      Since 1953,  73 square miles of natural marsh  has been  lost
 (Exhibit 13), 70% of it to open  water, the  rest  to impounded marshes,
 agriculture  and urban  habitats.  This figure represents  a loss of  26%
 of the total natural marsh area  of the basin.  The consequences  in
 natural  resources are  estimated  very roughly at  a  loss of 260,000  mt of
 organic  production,  77,000 birds per year,  19,000  fur pelts, and signif-
 icant impact on the shrimp and menhaden  fisheries.
                                 90

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 EXHIBIT 13:  CHANGE IN AREA OF HABITATS IN THE
CALCASIEU, LOUISIANA BASIN BETWEEN 1953 AND 1975
Habitat
Open Water
Nearshore Gulf
Natural Marsh
Saline
Brackish
Intermediate
Fresh
Altered Marsh
Ridge
Natural
Spoil
Agriculture
Rice
Non-rice
Pasture
Urban
Upland Forest
Beach
Swamp Forest
TOTAL
Area
1953
107.
156.
284.




23.

32.
9.

17-
5.
24.
3.
7.
3.
3.
678.
(sq.
52
00
30




91

45
58

03
18
41
28
50
51
42
09
miles)
1975
158.13
155.

8.
101.
78.
22.
37.

31.
12.

22.
6.
23.
8.
5.
3.
2.
678.
38

28
66
81
84
65

12
78

05
01
05
79
52
26
76
09
Net Area Gain
(Loss)
50.
(0.
(72.




13.

(1.
3.

5.
0.
(1.
5.
(1.
(0.
(0.
0.
61
62)
71)




74

33)
20

02
83
36)
51
98)
25)
66)
00
Net % Change
(1953-1975)
47.1
0.4
- 25.6




57.5

4.1
33.4

29.5
16.0
- 5.6
168.0
- 26.4
7.1
- 19.3

                      91

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REFERENCES

Bahr, L.M., J.W. Day Jr., T. Gayle, and C.S. Hopkinson.  1977.   "A Con-
  ceptual Model of the Chenier Plain Coastal Ecosystem of Texas  and
  Louisiana."  Center for* Wetland Resources, Louisiana State University,
  Baton Rouge, Louisiana.

Ehrenfeld, D.W.  1976.  "The Conservation of Non-Resources," Amer. Sci._
  64_(6) :648-656.

Forrester, J.W.  1971.  World Dynamics.  Wright Allen, Cambridge,
  Massachusetts, 142 pp.

Krewer, J.N. and S.W. Nixon.  1975.  "An Ecological Simulation Model of
  Narragansett Bay - The Plankton Community."  rn L.E. Cronin, ed. ,
  Estuarine Research, Vol. 1.  Academic Press, New York, p. 672-690.

Nichols, J.D., L. Viehmaw, R.H. Chabreck, and B. Fenderson.  1976.
  "Simulation of a Commercially Harvested Alligator Population in
  Louisiana," Louisiana Agricultural Experiment Station Bull. 691, 59 p.

Odum, E.P.  1971.  Fundamentals of Ecology.  W.B. Saunders Co.,  Phila-
  delphia, Pennsylvania.

Patten, B.C. ejt al.  1975.  "Total Ecosystem Model for a Cove in Lake
  Texoma."  In B.C. Patten, ed., Systems Analysis and Simulation in
  Ecology, Vol. 4, Academic Press, New York.

Wagner, F.H.  1969.  "Ecosystem Concepts in Fish and Game Management,"
  In G. Van Dyne, ed., The Ecosystem Concept in Natural Resource
  Management.  Academic Press, New York, p. 259-307.

Wiegert, R.G., R.R. Christian, J.L. Gallagher, J.R. Hall, R.D.H. Jones,
  and R.L. Wetzel.  1975.  "A Preliminary Ecosystem Model of Coastal
  Georgia Spartina Marsh."  In L.E. Cronin, ed., Estuarine Research,
  Vol. I, Academic Press, New York, p. 583-601.
                                 92

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   FATE AND EFFECTS OF CONTAMINANTS IN MARINE ECOSYSTEMS AND ORGANISMS
                             Douglas A. Wolfe*
     The general objectives of the National Oceanographic and Atmos-
pheric Administration's  (NOAA) energy resources project on fate and
effects are to identify and conduct laboratory and field research
studies that will facilitate the assessment of potential environmen-
tal impacts from petroleum releases in coastal marine ecosystems.
This effort does not have a site-specific orientation; instead, it
is focused on the fundamental processes that control the distribu-
tion, transport, and effects  (including physiological and ecological
aspects) of petroleum components, toxic metals, and other contaminants
in coastal ecosystems.  It is expected that the results of the
process-oriented studies conducted under this project will have broad
applicability to environmental problems in widely separated geographic
areas.

     This energy resources project represents only a small portion of
NOAA's overall effort on fate and effects of contaminants in marine
systems.  Within its various research laboratories, the National Marine
Fisheries Service  (NMFS) conducts a wide variety of marine environmental
research projects.  These projects are directed toward defining the envi-
ronmental requirements of fishery resource organisms and the effects of
environmental additions or alterations to those resources.  NOAA is also
managing for the Bureau of Land Management the outer continental shelf
environmental assessment program for environmental studies in support of
petroleum exploration and development on the Alaskan Continental Shelf.
In addition, two other energy resources projects are managed by NOAA:
environmental assessments of the Northern Puget Sound and the Buccaneer
oil field in the Gulf of Mexico.  A high degree of interaction occurs
throughout these various activities to minimize redundancy of effort
and to promote effective research planning in all aspects of NOAA's pro-
gram.
*    National Oceanographic & Atmospheric Administration, Environmental
Research Laboratories, Boulder, Colorado 80302.
                                93

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     The technical development plan (NOAA 1975) for the energy resources
project on fate and effects identified four major tasks to be under-
taken :

1.   Establishment of an NOAA national analytical facility for analysis
     of petroleum hydrocarbons and toxic metals in the marine envi-
     ronment

2.   Synthesis of existing information and identification of research
     needs on fate and effects of toxic metals and petroleum hydro-
     carbons in subarctic marine ecosystems

3.   Laboratory research on specific effects or transport processes
     involving petroleum hydrocarbons and toxic metals in marine
     ecosystems

4.   Experimental ecosystem research on ecological responses to
     additions of petroleum hydrocarbons.

     During the first year, the efforts of this project on fate and
effects were oriented mainly toward establishment of suitable in-house
analytical support capability for NCAA's overall environmental program
and the planning for future studies on fate and effects.  In addition,
laboratory studies were undertaken on the interactive effects of
petroleum and polychlorinated biphenyl (PCB)  exposure in fish and on
the interactions of petroleum and metals in seawater-sediment systems.
 NOAA NATIONAL ANALYTICAL FACILITY

     The NOAA National Analytical Facility  (NNAF) was established in
 August  1975 within the Northwest and Alaska Fisheries Center of NOAA's
 NMFS.   Procurement of equipment and recruitment of staff began imme-
 diately.  By March 1976, analytical work had been started by the full-
 time staff of four chemists under the direction of Dr. William MacLeod.
 At its  Seattle location, NNAF has the added advantage of direct
 association with a multidisciplinary research staff engaged in ex-
 perimentation on contaminant-induced changes in marine biota.  This
 interaction will promote the realistic evaluation of analytical results
 in terms of observed effects on marine organisms.

     The NNAF has six primary purposes:

 1.   Recommend the kinds of analyses needed to fulfill the objectives
     of environmental projects (e.g., in petroleum hydrocarbon analysis,
     should emphasis be placed on total hydrocarbons or on discrete
     fractions thereof?)

 2.   Undertake the examination and analysis of samples of biota, water,
     or sediments from both field and laboratory projects
                                94

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3.   Adapt  and,  where necessary,  develop methods required by current
     needs  of  NOAA programs

4.   Interpret data in relation to the biological and  ecological  im-
     portance  to the biota and place them in the appropriate perspective
     for  administrative decisions and actions

5.   Adapt  new methodology as objectives and needs of  NOAA programs
     change

6.   Provide contract liaison and quality control when analyses are
     conducted by other laboratories.

     Major  equipment acquisitions  consist of  a Finnegan 3200 gas
chromatograph-mass  spectrometer-data  system  (GC-MS), a scanning electron
microscope,  two  automated  gas  chromatographs, a  high-pressure liquid
chromatograph,  and  a fluorescence  spectrometer.   These acquisitions
supplement  an  extensive  inventory  of  equipment already available within
the Fisheries  Center.  With appropriate  sample collection and prepara-
tion techniques,  they will permit  the  reliable analysis of organic
compounds at less  than ppb levels.

     Considerable  effort was  invested  in training personnel on existing
techniques  and procedures,  including  revision and modifications where
necessary.   Much of the  analytical work  in the next few years will
involve hydrocarbon residues  in sediment and  tissue samples.   The NNAF
staff has studied  various  analytical procedures  for such samples, e.g.,
procedures  of  Warner (1976) ,  Rohrbach  and Reed (1975), and those under
consideration  by the Bureau of Land Management (1975).  After careful
consideration,  the  Warner  procedures were provisionally adopted for
training  and testing.   They have proven  to be generally convenient and
useful with  certain notable exceptions.   A major problem arose with the
use of teflon-lined centrifuge tubes  in  the alkaline tissue digestion
at 90C.  The  linings failed  to seal  reliably, causing serious losses of
moderately  volatile hydrocarbons.  This  problem was solved by digesting
overnight at lower  temperatures.   The  silica  gel  chromatographic clean-
up step also was  troublesome.   Mussel  and snail  tissue extracts effec-
tively plugged the  column.  Alternative  procedures are under investiga-
tion to alleviate  this problem.  Fortunately, our revision of the Warner
method does  not  require  separation of  the aromatics into two fractions
for GC analysis.   Our GC column techniques permit the aromatics to be
analyzed with  good  resolution  in a single  gas chromatogram.  Recoveries
of 85-90% for  the  examined  hydrocarbons  have  been obtained from spiked
control samples  at  a minimum  average concentration of 5 mg of individual
compound/g wet tissue.   Presently, the GC sensitivity is 0.25 mg
absolute amount  of  a single compound.  Analytical methods are detailed
in the report  produced by  NNAF under contract to NOAA's energy-related
Puget Sound project (MacLeod  et al. ,  1976).
                                 95

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     Studies undertaken by NNAF to date include the development and
application of analytical techniques for hydrocarbons in biological
samples of snails and mussels  (MacLeod et al_._, 1976, 1977).  In addi-
tion, NNAF performed the analyses for the experiments on hydrocarbon-
PCB interactions in salmon, which were also supported under this energy
resources project (Gruger et al., 1977a, 1977b).
FATE AND EFFECTS SYMPOSIUM

     On November 10-12, 1976, NOAA and EPA cosponsored a major sympo-
sium, held in Seattle and entitled, "Fate and Effects of Petroleum
Hydrocarbons in Marine Ecosystems and Organisms."  The symposium, which
was attended by approximately 350 persons from several countries, was
intended to provide a vehicle for documenting the current status of
research on the fates and effects of petroleum in the marine environ-
ment and for identifying areas most in need of future research.  Twelve
recognized experts were invited to present summary reviews on various
aspects of petroleum in the marine environment (see Exhibit 1).  In
addition, to provide a detailed status of current research, contributed
papers were solicited from the research community-at-large on this
topic.  Seventy-one abstracts were received in response to this solici-
tation, and 58 of these were selected by the symposium planning and
editorial committee for actual presentation.  Fifty-one manuscripts were
submitted for publication in the symposium proceedings, and 35 of these
were accepted by the committee.  The final proceedings volume  (Wolfe,
1977) includes the invited and the accepted contributed papers, as well
as the panel discussion, which took place on the last day of the
symposium.  During this discussion, the panel and the audience expressed
their thoughts on where the research priorities of the future ought to
lie.

     In the following paragraphs, I have attempted to tabulate the
opinions expressed during the panel discussion on research needs.  This
list is clearly not an exhaustive itemization of all the imaginable
research topics that will be of value to researchers and environmental
decision makers over the next several years.  Instead, it represents the
current thinking of a few, highly perceptive scientists who have been
heavily involved in research or research management in this topical area
for several years.  No attempt has been made to rank these suggested
research areas and recommended approaches.

1.   Acute toxicity bioassays should generally be de-emphasized in
     favor of longer term exposures and sublethal effects.  Earlier
     static bioassays should be repeated using flow-through systems
     to improve estimates of species' relative sensitivities.  Addi-
     tional species or life stages suspected to be highly sensitive
     should be tested.   Larval and juvenile stages still appear to
     be best candidates for research animals in effects studies.
     Petroleum effects on birds should be assessed under realistic
                                  96

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          EXHIBIT 1:  INVITED SPEAKERS AND TOPICS AT THE
           SYMPOSIUM ON FATE AND EFFECTS OF PETROLEUM
         HYDROCARBONS IN MARINE ECOSYSTEMS AND ORGANISMS
1.    Oil Spills in the Alaskan Coastal Zone - The Statistical Picture.
     J.R. Harrald, D. Boyd, and C.C. Bates, U.S. Coast Guard,
     Washington, D.C.

2.    Hydrocarbons in the Water Column.  D.G. Shaw, University of Alaska
     Institute of Marine Sciences, Fairbanks, Alaska.

3.    Oil Slick Movement in Alaskan Waters.  P.J. Kinney, Kinnetic Lab-
     oratories, Santa Cruz, California.

4.    Oil Slick Weathering.  C.D. McAuliffe, Chevron Oil Field Research
     Co., La Habra, California.

5.    Biodegradation of Aromatic Petroleum Hydrocarbons.  D.T. Gibson,
     University of Texas, Austin, Texas.

6.    Bioconversions and Metabolism of Petroleum Hydrocarbons.  D.C,
     Malins, NMFS Northwest and Alaska Fisheries Center, Seattle,
     Washington.

7.    Accumulation and Turnover of Hydrocarbons in Marine Organisms.
     R.F. Lee, Skidaway Institute of Oceanography, Savannah, Georgia.

8.    Food Web Transport of Petroleum Hydrocarbons.  J. Teal, Woods Hole
     Oceanographic Institute, Woods Hole, Massachusetts.

9.    Comparative Toxicities of Petroleum Hydrocarbons in Marine
     Organisms.  S.D. Rice, NMFS Auke Bay Fisheries Laboratory,
     Auke Bay, Alaska.

10.  Sublethal Effects of Petroleum Hydrocarbons, J. Anderson, Battelle
     Marine Research Laboratory, Sequim, Washington.

11.  Morphological Abnormalities Produced by Hydrocarbon Exposure.
     J.  Hawkes, NMFS Northwest and Alaska Fisheries Center, Seattle,
     Washington.

12.  Ecological Effects of Petroleum in Marine Systems.  A.P. Michael,
     University of Massachusetts Marine Laboratory, Gloucester,
     Massachusetts.
                                97

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     exposure conditions (both long- and short-term).   All bioassay
     experiments, whether acute or chronic, should use realistic dose-
     time exposure regimes (based on concentrations and compositions
     likely to occur in natural systems)  to assess potential effects.
     Uptake and turnover of compounds should be documented by thorough
     chemical analysis of tissues, and subsequent experimental and
     analytical emphasis should be on those specific compounds actually
     accumulated by the exposed organisms  (instead of on some measure
     of total hydrocarbons).

2.   Long-term effects to be studied should emphasize studies on:

        Behavior (feeding, reproductive, migrational)

        Carcinogenicity

        Cellular ultrastructure

        Chemosensory mechanisms

        Disease incidence and susceptibility

        Enzyme activities

        Growth

        Immune responses

        Mutagenicity

        Reproduction.

3.   Effects of petroleum components, other than hydrocarbons and inter-
     mediary metabolites of hydrocarbons, should be examined.  The
     relative toxic effects of various petroleum constituents should be
     assessed.  Past emphasis has been on hydrocarbons per se mainly
     because of the relative ease of analysis, not because they are the
     known direct causative agents in manifesting the effects.

4.   Potential transfer through food webs should be examined for hydro-
     carbon metabolites and nonhydrocarbon components of petroleum.
     Persistence of metabolites in organisms, which could be examined
     through the use of radiotracers, is a corollary problem.

5.   Various physical and chemical forms of contaminants/ including
     chemically complexed forms, should be tested for relative bio-
     availability, metabolism, and toxicity to marine organisms.  The
     natural occurrence and rates of formation of such complexes should
     be determined in marine systems.
                                98

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6.   Petroleum-sediment interactions should be characterized, and the
     transfer of sediment-associated petroleum constituents to organisms
     should be quantitated.

7.   Synergisms between different types of contaminants and between con-
     taminants and physical parameters  (e.g., temperature and salinity)
     should be examined.

8.   Field studies should be designed and conducted to corroborate lab-
     oratory results.  Such studies might involve  intentional perturba-
     tion under controlled conditions,  long-term studies of oil spills,
     and ecological and physiological studies of chronically polluted
     areas and areas of natural  seeps.  Field experiments require very
     careful and thorough planning to ensure that  causality of effects
     will be documented and that sampling frequency and intensity are
     adequate to demonstrate statistically a desired  level of effect.

9.   Models of contaminant cycling in the environment should be refined
     to include the kinetics of  physical and biological transport pro-
     cesses  (e.g., solution, evaporation, adsorption, bioaccumulation)
     and chemical  and  biological modification processes  (e.g., photo-
     oxidation, biodegradation).  Compositional changes in petroleum
     should be determined and  should serve as a basis for experimental
     design in effects studies.

     The foregoing suggestions and recommendations will be useful in
planning and evaluating future research.  However, detailed proposals
must be evaluated  in the context of work already underway at the time,
and in terms of the availability, competency, and  operational readiness
of investigators to perform the  work.

INTERACTIONS BETWEEN PETROLEUM AND
POLYCHLORINATED BIPHENYLS IN SALMON

     In marine environments, numerous types of contaminants exist to-
gether, including petroleum hydrocarbons and PCBs.  These xenobiotics
are expected to produce a variety of biological alterations in marine
organisms, such as biochemical and morphological changes in cellular and
subcellular systems, that could possibly influence their survival rate.
Studies of the effects of contaminant mixtures are important for 'judging
possible harm that may come to marine resources exposed simultaneously
to such compounds.

     A study was designed to delineate  nonlethal effects of chloro-
biphenyls and petroleum hydrocarbons on salmon  (Oncorhynchus species)
when mixtures of each xenobiotic were included in  the diet separately
and together (Gruger et al., 1977a).  Specifically, the research includ-
ed studies of accumulations and depurations of the test compounds, the
influences on an enzyme system (aryl hydrocarbon hydroxylases) associ-
ated with their metabolism, and  associated changes in cellular struc-
tures of key organs.
                                 99

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     Coho salmon  (Oncqrhynthus kisutch) and chinook salmon
 (o. tshawytscha) were exposed to two mixtures of pure chlorobiphenyls
and petroleum hydrocarbons, separately and together, at  1 ppm  and  5 ppm
of each mixture in Oregon moist pellets.  Chemical analyses of coho
salmon tissues indicated that the chlorobiphenyls accumulated  in the
liver, kidney, and muscle tissues; hydrocarbons were not detected  in
these areas.

     Microscopic examinations indicated that the chlorobiphenyls and
hydrocarbons had no effect on the morphology of gill tissue; however,
there was a possible dysplasia of the surface cells of skin.   The
effects of chlorobiphenyls on liver were variable; in one Chinook  salmon
there was severe vesiculation of hepatocytes, as well as whorled intra-
cellular membranes  similar to neoplastic cells in rats.  Chinook
salmon treated with both chlorobiphenyls and chlorobiphenyls and hydro-
carbons together exhibited mucosal sloughing in the intestine.  No
tissue damage was observed by light microscopy in fish receiving hydro-
carbons alone.  It is provisionally concluded that chlorobiphenyls and
hydrocarbons have a synergistic effect on the severity of sloughing of
 the intestinal epithelium and exudation of mucus.

     Aryl hydrocarbon  [benzo (a) pyrene] hydroxylase activities of
liver microsomes were compared among test and control salmon of both
species in order to determine whether alterations in enzyme activities
occurred in relation to the exposures (Gruger et al., 1977a; 1977b).
Optimal reaction conditions were established for measurements  of aryl
hydrocarbon hydroxylase activities in salmon hepatic microsomes; the
 conditions indicated differences compared to mammalian microsomes  and
differences in pH for the reactions for coho salmon compared to chinook
salmon.

     Induction of the aryl hydroxylase enzyme occurred in coho salmon
within 2 weeks of exposure to the hydrocarbons.  In addition,  tlje
induction was potentiated by the presence of the chlorobiphenyls,  which
by themselves had no effect on the enzyme activity.  A synergistic
effect was indicated for the test compounds on the induction of aryl
hydrocarbon hydroxylase in coho salmon livers.  In chinook salmon, the
activity of aryl hydrocarbon hydroxylase in hepatic microsomes was
depressed by the chlorobiphenyls and hydrocarbons administered, both
separately and together.  The results provide evidence indicating  that
different salmonid species respond differently to chlorobiphenyls  and
petroleum hydrocarbons in food in terms of the hepatic aryl hydro-
carbon hydroxylase system.
INTERACTIONS BETWEEN PETROLEUM AND TOXIC
METALS IN SEDIMENT-SEAWATER SYSTEMS
     Sediments constitute a very large reservoir for metallic elements
in aquatic environments, and the steady-state partition coefficients and
exchange rates depend upon redox potential, pH, and other chemical
                                100

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characteristics of the  sediments,  interstitial  water,  and overlying
water column.  Contamination  of  the  benthic  or  intertidal marine  sys-
tem by petroleum could  change this chemical  microenvironment,  thereby
affecting the distribution  and biological  availability of metals  to
demersal and benthic organisms.

     To measure the magnitude of this potential secondary impact  of
petroleum, various concentrations  of crude oil  will  be added to experi-
mental mixtures of marine sediments  and sea  water that have equilibrated
with respect to distribution  of  selected metallic elements.  After oil
impaction, the rate of  change of elemental concentrations in the  water
and the ultimate distribution of the metals  between  sediment and  water
will be measured.  Thus, the  effects of known compositions and concen-
trations of oil on the  exchange  rates and  partition  coefficients  of
metals between water and sediments will be determined.   The results of
this study will be correlated with existing  information on bioaccumula-
tion of metallic elements to  assess  the significance of petroleum con-
tamination with regard  to accumulation and toxicity  of metals  in  the
marine environment.  This experiment is currently in the initial  phase
of study, and results are not yet  available.
DESIGN OF  EXPERIMENTAL ECOSYSTEM STUDIES
     Carefully  controlled studies  of petroleum behavior and impacts
under  large-scale  experimental  conditions  in  relatively undisturbed
ecosystems provide the  most feasible means of assessing potential
petroleum impacts  at the  population or  ecosystem  level of organization
and  for  testing and verifying the  results  of  laboratory experiments
under  field  conditions.   Experimental releases of oil in an isolated
portion  of a larger study area  must, however, be  carefully designed to
approximate  closely the impact  of  oil on the  whole natural ecosystem.
In addition,  the experimental system must  be  well understood eco-
logically prior to initiation of experimental releases of petroleum.
It should be thoroughly characterized in terms of all those parameters
that might significantly  affect interpretation of data on a specific
hypothesized impact.

     Such a  characterization probably should  produce reasonable under-
standing of  species productivity,  variability in  community structure
and  diversity,  trophic  relationships, and  seasonal successionary
changes.  Processes affecting the  transport and distribution of petro-
leum should  also be characterized, including  water circulation, sedi-
mentation rates and distributions, and  air-sea interactions such as
evaporation  and aerosol formation.  Experiments should be designed to
document and measure both short-term effects  and  the effects of persis-
tent low-level  petroleum  exposure  in nearshore ecosystems, ranging from
the  subtidal littoral zone through the  intertidal zone to the adjacent
beaches  or wetlands.  These studies should include shore birds and mam-
mals and other  terrestrial trophic elements that  may become exposed to
the  petroleum.


                                101

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      The inherent complexity and natural variability of natural eco-
 systems require that studies of this type be designed with a great deal
 of care to ensure that:

     Sampling frequency and intensity are adequate to document hypothe-
      sized effects to within specified statistical limits

     The parameters selected for measurement are appropriate to demon-
      strate causality for a particular hypothesized effect.

      A major constraint limiting the ability to achieve adequate design
 of ecological perturbation studies is our lack of understanding about
 how the ecosystem functions naturally.  That is, what are the important
 causative interactions and how do they interrelate quantitatively?

      Given this lack of understanding, it becomes essential to limit
 the scope of experiments to those specific hypothesized impacts that we
 can realistically expect to test with a rigorous sampling design.   The
 use of experimental perturbation studies under natural field conditions
 should be restricted to those issues that cannot be addressed adequately
 under accidental oil spill conditions.  Thus, the gross effects of
 smothering on an oiled intertidal zone or the recolonization of a de-
 nuded shore by mussels are not questions warranting the expense of
 designing and conducting controlled field experiments on oil impacts.
 These dramatic effects can be readily quantified at the scenes of
 accidental oil spills (although sampling of such phenomena has rarely,
 if ever, been designed to provide additional insight on the sequence of
 causal mechanisms involved in the process).   On the other hand,  con-
 trolled spills, coupled with carefully conceived manipulations of the
 ecosystem, maV be the only means of verifying the relative importance
 of specific food-chain relationships in transferring contaminants
 between organisms under actual field conditions, or for separating the
 effects of predation and pollutant toxicity on population stability of
 a given species.  The controlled experimental spill provides realism in
 the introduction, the natural transport pathways, the chemical composi-
 tions, etc., of petroleum in a marine system.  Concurrently, it provides
 a stage for conducting ecological manipulations that can document
 mechanistic causality for selected processes of interest.

      Numerous conferences on biological effects have decried the valid-
 ity or applicability of laboratory results to field situations and have
'almost casually recommended controlled field experiments as the logical
 approach to achieve the desired realism.*  A workshop sponsored by
 Exxon (1974) also espoused the controlled field experiment as a means
 of studying chronic effects of oil in marine systems.  It is clear from
 *    See,  for example,  recommendation no.  8 above,  which was put forth
 on several occasions at the recent fate and effects symposium sponsored
 under this project (Wolfe,  1977).


                                102

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the foregoing discussion that  the  realities  of  ecosystem complexity  and
natural variability severely restrict  the real  utility of controlled
field experiments and demand a high degree of planning and experimental
design prior to implementation.

     As a first step toward the necessary planning, NOAA (through a
contractor) is conducting design feasibility analyses  for various ex-
perimental approaches and ecological hypotheses that might be appropri-
ately tested in field perturbation experiments  with petroleum.  Results
of the study should be available this  year.

     Future work under this project will  continue to be  directed toward
understanding specific processes and mechanisms affecting the movement,
distribution, and effects of contaminants in marine systems.  It is
expected that, when adequate design criteria are developed, controlled
field experiments will be utilized interactively with  laboratory
experiments for this purpose.
REFERENCES

Bureau of Land Management.   1975.   Draft  Summary Recommendations of the
  Trace Metal and  Hydrocarbon  Seminars, Aerospace Corporation Rpt. No.
  ATR-76(7504)-5.

Exxon Production Research  Co,   1974,   "Research Needs to Determine
  Chronic Effects  of  Oil on  the Marine Environment," Proc. of a Work-
  shop, November 4-6,  1974,  Houston, Texas.

Gruger, E.H., Jr.,  J.W. Hawkes, and D,C.  Malins.  1977a.  Effects of
  Chlorinated Hydrocarbon  Residues  on  the Bioaccumulation and Toxicity
  of the Petroleum Hydrocarbons, Final Rpt. for Work Unit No. 3-3-2,
  Interagency Agreement EPA-IAG-E693,  NOAA Energy Resources Project.

Gruger, E.H., Jr.,  M.M. Wekell, and P.A.  Robisch.  1977b.  "Effects of
  Chlorinated Biphenyls and  Petroleum  Hydrocarbons on the Activity of
  Hepatic Aryl Hydrocarbon Hydroxylase of Coho Salmon (Oncorhynchus
  kisutch) and Chinook Salmon  (O. tshawytscha)."  In;  D.A. Wolfe, ed.,
  Fate and Effects of  Petroleum Hydrocarbons in Marine Ecosystems and
  Organisms, Pergamon  Press, New York:  323-331.

MacLeod, W.D., Jr., D.W. Brown, R.G. Jenkins, L.S. Ramos, and V.D.
  Henry.  1976.  "A Pilot  Study on  the Design of a Petroleum Hydro-
  carbon Baseline  Investigation for Northern Puget Sound and Strait of
  Juan de Fuca," Natl. Ocean.  Atmos, Admin. Tech. Memo.  No. ERL-MESA 8,
  Environmental Research Laboratory, Boulder, Colorado,  18 pp.
                               103

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MacLeod, W.D., Jr., D.W. Brown, R.G. Jenkins, and L.S. Ramos.  1977.
  "Intertidal Sediment Hydrocarbon Levels at Two Sites on the Strait of
  Juan De Fuca."  In:  D.A. Wolfe, ed., Fate and Effects of Petroleum
  Hydrocarbons in Marine Ecosystems and Organisms, Pergamon Press, New
  York:  385-396.

National Oceanic and Atmospheric Administration.  1975.  Project
  Development Plan.  Fate and Effects of Petroleum Hydrocarbons and
  Selected Toxic Metals in Selected Marine Ecosystems and Organisms,
  NOAA Energy Resources Project, 31 pp.

Northwest Fisheries Center.  1976,  "NOAA's National Analytical Facility
  Established at Northwest Fisheries Center,"  NWFC Monthly Report,
  Seattle, Washington:-  11-19.

Rohrback, B.C. and W.E. Reed.  1975.  "Evaluation of Extraction Tech-
  niques for Hydrocarbons in Marine Sediments," Inst. of Geophys. and
  Planet. Phys., Publ. No. 1537, University of California at Los
  Angeles, Los Angeles, California, 23 pp.

Warner, J.S. 1976.  "Determination of Aliphatic and Aromatic Hydrocar-
  bons in Marine Organisms," Anal. Chem. 48  (3):  578-583.

Wolfe, D.A., ed.  1977.  Fate and Effects of Petroleum Hydrocarbons in
  Marine Ecosystems and Organisms, Pergamon Press, New York, 4 pp.
DISCUSSION

     From the floor:  Could you give us a rationale for your choice of
hydrocarbon levels and PCB levels in your salmon experiment?

     Dr. Wolfe:  We may not have conducted adequate studies on the
threshold levels of PCB and petroleum required to induce the enzymatic
activities.  The selected concentrations are representative of total
hydrocarbon levels found in marine organisms under natural environmental
conditions.  The PCB levels are still high.  Again, the purpose of the
experiment was to determine whether there is a synergistic interaction
between the two components, so we wanted to emphasize the effects of
the PCBs.

     Dr. Lefcort:  Would you elaborate on your plans for these field
studies?

     Dr. Wolfe:   In the beginning, we thought that we could character-
ize a natural marine system to an extent that we could then hypothesize
certain effects of petroleum and conduct a deliberate spill to verify
or refute those hypotheses.  Consequently, the project was oriented
towards deliberate spills, and the ongoing feasibility analyses are
still directed toward deliberate spills.
                               104

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     NOAA is currently undertaking  a  spills  of  opportunity program
funded under the outer continental  shelf  environmental  assessment
program.  Under this program,  NOAA  has  attempted to  identify  specific
research questions that might  be  feasible under opportunistic condi-
tions.  We have assembled  four spilled-oil research  teams, which are
able to respond on the basis of a midnight phone call to  a spill that
occurs almost anywhere at  anytime.

     So far, the objectives that  have been put  forth for  opportunis-
tic spills are confined to questions  of physical transport and physical
behavior of oil in a marine environment.   At this time, we have not
designed any biological experiments that  we  considered  feasible.

     Dr. Barrientos;  Sometime last November, Craig  Hooper put together
the environmental impact  statement to do  a control on this experiment.
Are these field experiments related to  that  at  all?

      Dr. Wolfe:   A  deliberate spill was designed under  the outer
continental  shelf program. This  program  reached the point of a
license request draft to  be presented to  EPA.   The proposal was to
assess the physical parameters of petroleum  behavior in the marine
environment  in the  Gulf of Alaska.  That  draft  was undergoing review
when  the Argo Merchant went aground.  Because of several  different con-
straints resulting  from  that  disaster,  that  proposal has  been postponed.
                                 105

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         LABORATORY AND FIELD STUDIES ON THE LONG-TERM EFFECTS
      OF PETROLEUM HYDROCARBONS ON BENTHIC MARINE INVERTEBRATES
        J.W. Anderson, R. Riley, R.M. Bean, J.W. Blaylock, and S.L. Kiesser*
     As a result of oil production  from the Alaskan outer continental
shelf, the Pacific Northwest is one of the areas of the United States
most likely to receive increased inputs of petroleum hydrocarbons  (PHC)
from transport, port operations, and refining.  There is the possibility
that exploration on the coasts of Oregon and Washington will indicate
the potential for drilling; thus, production-related inputs are pos-
sible.  Present and future activities associated with this energy source
necessitate research on the potential impacts of oil in the Pacific
Northwest marine environment.  Although the results of recent studies
on the fate and effects of PHC in the marine environment have strength-
ened our understanding of some aspects of oil pollution, we are not yet
to the point at which an accurate prediction of the effects of acute or
chronic oil inputs can be made.

     To define more clearly the potential for petroleum to exert signif-
icant long-term environmental effects in marine sediments, studies on
the fate of crude oil in sediments  are being conducted.  These inter-
disciplinary studies involve aspects of physical organic chemistry,
marine biology, and microbiology.   Their principal objectives are to
determine the rates and mechanisms of degradation of petroleum in the
marine environment, to determine which component types are persistent,
and to correlate such information, where possible, with observed ef-
fects on the marine biota.

     To meet the analytical objectives, considerable effort has been
directed toward development of analytical methods for the separation,
identification, and quantitation of classes of compounds and individual'
compounds that comprise saturate and aromatic fractions of Prudhoe Bay
crude oil.  Analytical methodologies developed on Prudhoe Bay crude oil
have been appropriately modified for analysis of petroleum compounds in
sediments, tissues, and laboratory exposure systems.

     In this paper, we report on those methods that are being used to
meet some of the above needs.  These techniques are being used to
*    Battelle Marine Research Laboratory, Route 5, Box 1000, Sequim,
Washington 98382.
                               107

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separate, identify, and quantitate the components of Prudhoe Bay crude
oil and to quantitate these components in sediment samples contaminated
with the oil as a part of laboratory and field depuration studies.  The
methods used in these studies employ the techniques of liquid-column
chromatography, capillary-columm chromatography, gas chromatography-mass
spectrometry  (GC-MS), high-performance liquid chromatography  (HPLC), and
infrared spectroscopy (IR).

     To provide the scientific data base necessary to evaluate the im-
pact of oil in the marine environment, large-scale programs incorpo-
rating a "team effort" are required.  Marine biologists, with expertise
in different fields (e.g., ecology, physiology, biochemistry), must work
closely with analytical chemists, and each should utilize the best
available techniques while attempting to develop even better approaches.
From a review of the literature on fate and effects of oil, much of
which was updated at a recent symposium in Seattle (Fate and Effects of
Petroleum in Marine Ecosystems and Organisms, ed. D.  Wolfe, NOAA), it
appears to us that at least four major questions remain to be answered:

1.   When exposed to oil, which specific PHCs are retained longest and
     to the greatest extent in organisms and sediment?

2.   Are sediment-sorbed PHCs available to organisms feeding and/or
     burrowing in the substrate, and is there biomagnification?

3.   As a result of oil toxicity or chemical interference, are larval
     benthic organisms eliminated from contaminated sediments (lack of
     recruitment), and what are the time and concentration factors in-
     volved?

4.   What is the threshold PHC level at which long-term exposure no
     longer alters marine communities nor reduces the capability of
     sensitive organisms to grow and reproduce normally?

     We believe that the analytical developments and biological ap-
proaches discussed in this paper provide the scientific data base neces-
sary to answer all or part of these questions.
MATERIALS AND METHODS

     We have categorized this section according to those analyses that
involve analytical chemistry and those that are biologically oriented.

     Analytical Chemistry

     In this section, we describe the analysis of PHCs in the flowing
sea water system, the analysis of Prudhoe Bay crude oil, field and lab-
oratory studies of depuration of oil in sediment, analysis of total oil
in sediments, and field and laboratory studies of separation and quanti-
tation of compounds in sediments.


                                IDS

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     Analysis of PHCs.   Investigations of the toxicity of crude  and
refined oils in  sea  water have been underway at Battelle-Northwest's
Marine Research Laboratory for several years.  A principal means of
carrying out these studies has been the use of continuous flow oil
exposure apparatus developed  at the Battelle-Northwest Marine  Research
Laboratory  (Vanderhorst  et al.,  1976a).  To determine  total oil  concen-
trations and to quantitate individual hydrocarbons at  the concentrations
currently being used,  two separate analytical techniques  have  been
developed and applied.   The first technique involves extraction  of
petroleum-contaminated sea water samples with helium,  and injection  of
the helium  extract into  a gas chromatograph for determination  of vola-
tile hydrocarbon component concentrations.   This method is about three
orders of magnitude  more sensitive than conventional methods.  Bean  and
Blaylock  (1977)  recently described it in detail, and Vanderhost  et al^
 (1976a) applied  it in  a  study of No.  2 fuel oil toxicity  utilizing oil
concentrations of less than 200 ppb.

     The second  analytical method allows the characterization  of low
concentrations of oil  in sea  water and involves the  use of resin ad-
sorption.   Approximately 10 1 of sea water  from the  flow-through oil
exposure apparatus are pumped through 3/8 in.  columns  containing XAD-2
macroreticular resin.  The resin has been shown to efficiently adsorb
the oil components from  the sea water even at low concentrations.  Re-
covery of oil  from a sea water suspension containing 115  ppb Prudhoe Bay
crude oil was  92.4  1.7% efficient,  based  on triplicate  determinations.
Analysis of the  adsorbed oil  is accomplished by eluting the  column with
carbon tetrachloride and determining the IR absorbence of the  eluate.
It has been found that the analytical columns can be regenerated at
least nine  times" (by washing  with methanol,  then water) before new
resin must  be  employed.

     Analysis  of Prudhoe Bay  crude oil.  The separation of Prudhoe Bay
crude oil into saturate  and aromatic compound fractions was  adapted
from a silica  gel chromatography technique  described by Warner (1976)
and modified by  MacLeod  _^t al.  (1976) .   Two fractions  (saturate  and
aromatic) were collected.  Each fraction with its appropriate  internal
standard was concentrated and analyzed by gas chromatography on  a 30 m
SE-30 glass capillary column  (J & W Scientific)  programmed from  an ini-
tial temperature of  70  to 250 at a rate of 4/min.   Compounds  in the
saturate and aromatic fractions were identified by peak enhancement
analysis with  known  standards.   A Hewlett Packard 5980A quadrapole mass
spectrometer operating in the argon chemical ionization mode and employ-
ing single  ion monitoring was used to determine approximate retention
time ranges for  specific compound types within the aromatic fraction.

     Field  and laboratory studies of depuration of oil in sediment.  A
surface layer  of oil (except  controls)  was  deposited in trays  containing
coarse beach sediment at a level of 4% of the sediment weight.   Forty-
eight hr of tidal flushing (four cycles)  were used to  remove excess  oil
from the surface of  the  sediments.  After the 48 hr  flushing period,
the trays were placed in the  field.  Since  the initiation of the


                                109

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experiment on April 17, 1976, samples were collected and frozen approx-
imately every 2 months.  Samples were treated and analyzed as described
below.

          Depuration of oil in sediments  (laboratory study).  The  labor-
atory study was initiated early in 1976 by introducing the fine sediment
mixed with 1% fresh Prudhoe Bay crude into an aquarium.  An additional
aquarium, serving as a control, was charged with fine-textured sediment
free of Prudhoe Bay crude.  The depth of sediment was approximately
4 in., and fresh sea water was allowed to continuously flow over the
sediments in both aquaria.  Triplicate grab samples were removed and
frozen at intervals spanning 180 days.  Hexane and CCl^ extracts of the
sediments were analyzed by IR and HPLC.

          Analysis of total oil in sediments by IR.  Concentrations of
total oil in sediments were determined on carbon tetrachloride extracts
of sediments with a Beckman Acculab 6 spectrophotometer by comparison
of the amount of absorbance obtained at 2930 cnr1 to a calibration curve
(API, 1958).

          Separation and quantitation of compounds in contaminated
sediment  (field study).  Triplicate 20 g samples of sediment were
extracted for 24 hr in glass bottles containing teflon-lined caps, 20 g
of anhydrous sodium sulfate, and 50 m of hexane (Burdick & Jackson).
Appropriate amounts of each sample were concentrated to 1 m and column-
chromatographed according to the method of Warner (1976).  Saturate and
aromatic fractions were analyzed by capillary column chromatography as
previously described, employing 2,6,10-trimethyldodecane and hexamethyl-
benzene as internal standards.  Peak areas and internal standard cali-
brations were determined with an Autolab Systems IV Computer integrator.

          Total oil and separation and quantitation of classes of com-
pounds from oil-contaminated sediment (laboratory study).  Three yfc of
hexane extracts of sediment obtained as previously described were
analyzed by HPLC, utilizing three series-coupled 1/8 in.  x 2 ft Durapak
oxypropionitrile on porasil C columns (Waters Associates),  using hexane
as mobile phase and 2.0 m/min flow rate.   The columns were previously
validated with the model compounds dodecane, benzene, naphthalene, and
anthracene to demonstrate separation of hydrocarbons into saturate,
monoaromatic, diaromatic, and polyaromatic fractions.  The quantitative
analysis of the aromatics in sediment samples was performed with a
calibration curve from a Prudhoe Bay crude aromatics fraction prepared
according to an ASTM method (Vanderhorst et al., 1976a).   Saturate
fractions were detected with a Waters R-401 differential refractometer,
and the aromatic fractions were detected with a Schoeffel variable
wavelength ultraviolet (UV)  detector set at 221 urn.

     Biological Analysis

     In this section, we describe our flowing sea water and flowing
exposure systems.
                                110

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     Flowing exposure  system.   Our flowing sea water system for exposing
organisms to low levels  of soluble plus finely dispersed PEC has been
previously described by  Vanderhorst et al. (1976a) .   Prudhoe Bay crude
oil was used as the source o'f  contamination,  since  it is likely to  be
one of the primary products delivered to this area,  and other research-
ers in Alaska and the  Pacific  Northwest are utilizing the same oil  in
their research.  The hydrocarbon delivery system was adjusted to deliver
0.22-0.24 ppm total hydrocarbons,  determined by IR,  as described above.

     A mixture of intertidal sediments (3 cm and less in size)  was  col-
lected at mean low tide  for transfer into three fiberglass exposure
tanks  (90 x 40 x 30 cm) .  One  of the tanks received  clean flowing sea
water, while the other two received a mixture of sea water and an oil-
in-water dispersion prepared in the continuous-flow  apparatus (rate of
1.5
     Anonyx  nugax (Amphipoda :   Crustacea)  cultures of 100  individuals
were introduced  into the control tank and  one of the exposure  tanks on
October  4, 1976.   Uptake of total naphthalenes and long-term effects of
low-level  contamination on growth and reproduction were monitored.  A
1 mm screen  was  fastened over  the standpipe of the control tank  to pre-
vent the araphipods from escaping.  Outfall baskets were constructed on
exposure tanks to eliminate the buildup of a surface oil sheen,  while
still  retaining  the animals.  A second exposure tank was stocked with
100 amphipods  4  weeks later as a replicate.  Uptake of total naphtha-
lenes  in the tissue of exposed amphipods was measured after 7, 10, and
14 weeks by  the  UV spectrophotometric technique described  by Neff and
Anderson (1975) .   Effects on growth were determined by total length and
wet weight measurements .

     Sediment  exposure systems.  Using a system previously described by
Anderson e_t  al.  (1977) , peanut worms (phylum Sipunculida)  were exposed
to oil-contaminated sediment in the laboratory.  Sediment  boxes, made
of PVC or  fiberglass with bottoms covered  by fiberglass mesh, were
used to  hold sediment that had either been oiled on the surface  or mixed
thoroughly with  oil.  These boxes and the  burrowing worms  (Phascolosoma
agassizii) were  held up to 2 weeks in flowing clean sea water  and simu-
lated  tidal  fluxes periodically drained the substrate via  the bottom
mesh.  Analyses  were conducted during the  experiments to determine naph-
thalenes in  the  sediment and tissues and total hydrocarbons in the sedi-
ment.

     A similar approach has been utilized  in field experiments,  involv-
ing oiled  sediment and three species of benthic organisms.  Sipunculids
and two  clam species  (Macoma inquinata and Protothaca staminea)  were
placed in  boxes  containing either clean or oiled sediment  (622 ppm total
hydrocarbons by  IR) and these  were placed  in the intertidal zone for 40
days.  After 40  days, tissue samples of each species were  analyzed by
GC, following  the methods of Warner (1976) .
                                 Ill

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     Beginning in March 1976, sediment boxes containing clean and oil-
contaminated substrate were placed in the intertidal zone on the sand
spit east of the Sequim laboratory.  The first two installations (March
and April, 1976) consisted of six boxes each, four of which were con-
taminated by Prudhoe Bay crude oil via surface coating.  Using a tidal
flux system in the laboratory, a layer of oil (4% vertical velocity
sediment) was allowed to impact the surface for about 1 hr, before a
series of high  (above surface) and low (below the box) tides were initi-
ated.  After 48 hr (four tidal flushes) in the laboratory during which
excess oil was floated off the sediment,  the boxes were transferred to
the field site and installed flush with the natural sediment.  A third
installation in November 1976 was treated in a similar fashion, but
sediment was mixed thoroughly in a fiberglass-coated cement mixer
instead of receiving oil on the surface only.

     The first installation has now been in place for nearly 1 year,
during which numerous samples have been taken for analyses of specific
hydrocarbon composition and presence of recruited biota.  Methods of
chemical analyses are discussed above and the results of initial chem-
ical characterization by capillary gas chromatography  (Cap-GC)  and GC-
MS are presented below.  Analyses of benthic organisms found in the
sediment samples are still underway, but the numbers of two prominent
bivalves present in control and exposed sand in "June and August 1976
will be discussed.  Cooperella subdiaphana and Mysella tumida  were
selected in this preliminary report, since they clearly represent re-
cruited organisms and not just transient inhabitants of the sediment.
In the future, we will be able to report the number of species and in-
dividuals in each box at several time intervals over a period of years.
It is important to note the significance of being able to correlate
detailed and quantitative hydrocarbon analyses with the abundance and
diversity of benthic organisms over an extended period of time.
RESULTS
     We describe below our results for the above methods.

     Analytical Chemisitry Results

     The results of our analyses of the flowing exposure system,
Prudhoe Bay crude system, depuration of oil in sediments, are summarized
in the following paragraphs.

     Flowing exposure system.  Using the He-GC technique for determina-
tion of monocyclic aromatics, the flowing oil delivery system has been
characterized as shown in Exhibit 1.  Analyses on December 17, 20, and
21 show the remarkable consistency of component concentrations in our
exposure system.  As reported by Rice e_t al.  (1977) , monoaromatics tend
to remain in cold sea water to a greater extent than has been observed
in laboratory studies or field investigations in temperate waters.  The
low molecular weight compounds represent a significant proportion of


                                112

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the hydrocarbons  to which organisms are exposed in the system,  and it
is likely that  this would be true for chronic and acute inputs  to the
marine environment of  the Pacific Northwest.   It should be noted that
the total hydrocarbon  (IR)  values listed at the bottom of Exhibit 1 do
not necessarily relate to totals of low molecular weight compounds.
In fact, only a small  percentage of the latter hydrocarbons are detected
by IR, so these should be considered independent analyses.  A realistic
grand total would be somewhat less than the sum of these two separate
analyses.

     Resin column extractions of 10  samples from exposure water were
conducted from  September 14, 1976, to January 14, 1977, and are still
in progress.   Concentrations of total hydrocarbons were determined for
the same three  exposure tanks during this period.  Analyses shown in
Exhibit  2 have  been sorted into three time intervals,  as flow rates
were altered to suit our experimental requirements.  Tanks 4 and 5 were
calibrated to deliver  the same concentrations,  except  for the period of
October  20, 1976, to November 1, 1976.   Although only  a slight  decrease
in concentration  of Tank 4 was produced after October  19,  1976,  a major
decrease in Tank  6 was intentionally produced on October 20,  1976.   The
consistency within a given time period in a specific tank was very high,
as shown by the standard deviations.  One-way analyses of variations
 (ANOVA)  among tanks at each time interval demonstrated that all  con-
centrations were  equal during the first 35 days,  but later adjustments
produced significantly higher concentrations  in Tanks  5 (12-day period)
and 6  (last 82  days).   Without considering the adjustments,  the means
of Tanks 4 and  5  were  relatively close over the entire 117-day  period.
It is important to remember the consistency of the system and the levels
present  in Tanks  4 and 5, since these contained the amphipod cultures
to be discussed below.

     Analysis  of  Prudhoe Bay crude oil.   Chromatographic separation  of
Prudhoe  Bay crude oil  produced a saturate and an aromatic fraction.
Compounds that  were identified in the saturate fraction by gas-capillary
column chromatography  included the straight-chain hydrocarbons  from
C1:]_ to C26, pristane,  and phytane.  Pristane  and phytane were baseline
resolved from  heptadecane and octadecane (see Exhibit  3).   Identifica-
tion and quantitation  of compounds in the aromatic fraction have proved
more difficult  because of the lack in the availability of commercial
standards and  because  many are present in Prudhoe Bay  crude oil at much
lower concentrations relative to the saturate compounds.   Identifica-
tions that have been made are listed in Exhibit 4. Many compounds in
the aromatic fraction  were categorized by GC-MS according to compound
types, i.e., monomethyl-, dimethyl-, and trimethylnaphthalenes,
monomethyl-, C2-, and  C3-phenanthrenes,  etc.   Thus, the GC-MS informa-
tion has enabled  us to estimate the concentration of hydrocarbon types.
                                 113

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 EXHIBIT 1:   ANALYSIS OF  LOW MOLECULAR WEIGHT HYDROCARBONS  IN THE FLOWING
    SEA WATER SYSTEM RECEIVING PRUDHOE BAY CRUDE  OIL  [All values are
     expressed as pg/1  (ppb)  and water temperature was 8   0.2C.]

Petroleum Hydrocarbons
Benzene
Toluene
Ethyl Benzene
m and p Xylene
o Xylene
Trimethyl Benzenes
Saturates 
-------
        EXHIBIT  2:   TOTAL  HYDROCARBONS  (ppm)  IN THE  FLOWING SEA WATER
               SYSTEM AS DETERMINED BY IR  ANALYSES OF COMPOUNDS
               SORBED TO RESIN COLUMNS RECEIVING 10 H OF  WATER
                              TANK #4
TANK #5
TANK #6
Sample Interval
9/14/76-10/19/76
10/20/76-11/1/76
11/5/76-1/14/77
9/14/76-1/14/77
No.
Days
35*
12'1'
70f
117
No.
Samples M S.D.
17 0.31 0.141
6 0.23 0.032
16 0.16 0,055
39 0.24 0.126
No.
Samples M S.D.
17 0.22 0.084
6 0.37 0.045
16 0.16 0.045
39 0.22 0.032
Mo.
Samples
17
5
16
38
M S.D.
0.24 0.10
0.40 0.077
0.32 0.126
*0.30 0.122
*   Concentrations in the three tanks not significantly different  (p<0.05, one-way ANOVA).
t   Concentrations in the three tanks different (p<0.001, one-way ANOVft).

-------
               Jtk
                    Us
                      14
                                  ^17
                                       Pin
                                           19
EXHIBIT  3:  GAS CAPILLARY CHROMATOGRAM OF SATURATE  FRACTION
   ^11     * ^28'  saturate straight chain hydrocarbons,
  Pr = pristane,  Ph = phytane, Is = 2,6,10-trimethyl-
  dodecane  internal standard)
GAS CAPILLARY CHROMATOGRAM OF AROMATIC FRACTION
EXHIBIT 4:
Key:
    Naphthalene                       I.
    2-Methylnaphthalene               J.
    1-Methylnaphthalene               K.
    1-Ethyl '+ 2-Ethylnaphthalene      L.
    2,6 + 2,7*-Dimethylnaphthalene    M.
    1,3 + 1,6-Dimethylnaphthalene     N.
    l,7*-Dimethylnaphthalene          Is.
    1,4 + 2,3 + 1, 5-Dimethylnaphtha-
    lene
                              1,2-Dimethylnaphthalene
                              2,3,6-Trimethylnaphthalene
                              Phenanthreiie
                              Methylphenanthrenes
                              C -Phenanthrenes
                              C -Phenanthrenes
                              = Hexamethylbenzene
                                internal standard
*   2,7 and 1,7 are suggested structures based on published  litera-
ture on the law resolution gas chromatographic separation  of
dimethylnaphthalenes.
                             116

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     Depuration of oil  in  sediments.   The  above  technique was applied
to the field study of the  depuration  of  the  components of Prudhoe Bay
crude oil from a coarse beach  sediment.  The concentrations of compounds
in the saturate and aromatic fractions from  sediment taken from the day
of installation are listed in  Exhibit'5.   Dodecane, naphthalene, and
phenanthrene were selected as  representative saturate and aromatic com-
ponents of oil, and their  rate of  depuration in  the sediment was deter-
mined over a period of  7 months.   The results are  shown  in Exhibit 6.
Only 2.1%, 1.0%, and 2.6%  of the original  dodecane, naphthalene, and
phenanthrene, respectively, remained  in  the  surface portion of the sedi-
ment after 7 months of  exposure of the sediment  in the beach environ-
ment.  These results were  in excellent agreement with values of total
oil obtained on the same samples.

     The retention time of Prudhoe Bay crude as  a  function of total oil
and compound class was  determined  on  a fine-textured sediment under
anaerobic conditions.   We  used the method  employing IR and HPLC.  The
results of extraction of the sediment with carbon  tetrachloride followed
by IR analysis are shown in Exhibit 7.   These data were  based on anal--
yses of three subsamplings from each  of  two  sample replicates for each
time period.  The variance experienced among subsamples  of the same jar
 (coeff.*-3.7%) was less than the total variance  (coeff.  -5.8%).  These
data obtained for oil concentration indicate that  the rate of depuration
of oil from fine, undisturbed, anaerobic sediments can be very slow, on
the order of several years.

     Samples of hexane  extract of  the same sediment contaminated with
oil when analyzed by HPLC  (analysis time ^20 min)  produced a saturate
fraction and a monoaromatic/diaromatic fraction, which was approximately
80% resolved.  The data for the aromatics  analysis are also shown in
Exhibit 7.  The plot suggests  slow depuration of these components, but
at a rate somewhat faster  than the total oil.  Correlation of the data
with the linear regression (dashed line, Exhibit 3) was  better (r2 =
0.44 vs 0.31) than for  the total oil  curve.

     Results of Biological Analysis

     In the following paragraphs,  we  summarize the results of our anal-
yses of the flowing exposure system and  field exposure of oiled sedi-
ments .

     Flowing exposure system.  As  shown  in Exhibit 8, the amphipod
Anonyx~nugax accumulated a significant amount of naphthalenes during
constant exposure to the Prudhoe Bay  crude oil extract.  Considering
the individual variations, exhibited  by  the  standard deviations, the
animals reached a threshold of accumulation  by 7 weeks of exposure.
There was no significant increase  over this  level  of contamination at
the 10- or 14-week interval.   From additional unpublished data (Anderson
and Kiesser) , there is  a strong trend for  the levels of  specific com-
pounds in the tissues to change  from  approximately equal distribution
of naphthalene  (N) , methylnaphthalenes (MN), and dimethylnaphthalenes


                                117

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EXHIBIT 5:  CONCENTRATIONS OF SATURATE AND AROMATIC
   HYDROCARBONS (ppm) IN SEDIMENT PROM THE DAY OF
       INSTALLATION OF THE FIELD EXPERIMENT
Concentration
Saturates (ppm)
CLI
c 13
C m
c 15
c 1S
Cj7
13
16
17
17
19
18
17
Pristane 10
C lg
16
Phytane 3
C19
C2Q
C21
C22
C23

C2s


18
16
17
16
15
13
n
10

.34
.11
.79
.44
.39
.84
.03
.74
.20
.05
.79
.40
.56
.02
.18
.80
.71
.21

i
~
~
7
7
~
7
7
7
7
7
7
+
+
7
7
7

1.
1.
1.
2.
2.
3.
2.
1.
3.
1.
3.
2.
3.
3.
2.
2.
2.
1.

21
53
82
39
94
00
92
95
22
43
00
88
09
37
65
34
47
97

Total saturates
measured 274








MN =
OKN =
TMN =
MP =








.61

















42








.32








Methyl naphthalene
Dime thy! naphthalene
Trimethylnapthalene
Methyl phenanthrene
Aromatics
Naphthalene
2-MN
1 -MN
Total

1 -ethyl + 2 -ethyl
Naphthalene

2,6 + 2,7-DMN
1,3 + 1,6-DMN
1 ,7-OMN
1,4 + 2,3 + 1,5 OMN
1 ,2-DMN
Total

TMM-1
TMN -2
TMN -3
TMN -4
2. 3, 6 -TMN
TMN-5
Total
Phenanthrene
MP-1
MP-2
MP-3
MP-4
Total
C2-phenanthrene-l
C2-phenanthr=ne-2
C2-phenanthrene-3
C2-phsnanthrene-4
Total
Total aroma tics
measured
Concentration
(ppm)
7
14
10
24


6

9
9
7
9
1
42

3
3
5
4
2
5
28
3
2
2
3
3
12
L.
2
2
14

140
.20
.31
.51
.32


.93

.52
.93
.31
.14
.60
.68

.80
.65
.59
.25
.08
.99
.44
.05
.74
.34
.94
.49
.53
.36
.56
.47
.70

.35

+
7
7


*_

+
7
+
7
0.
3.
2.
'5.


0.

0.
0.
0.
0.
77
10
90
32


35

75
94
64
72
+ 0.45
4-

+
+
7
7
7
7
-h
1

7
7
^-
I-
7
-t-
7


2.

0.
0.
1.
0.
0.
0.
6.
0.
0.
0.
1.
1.
4.
.
0.
0.
0.
3.

23
74

64
55
n
75
24
77
10
85
56
40
71
67
31
56
68
44
37
52

.96
                       118

-------
  16

  14
I

1 I?
O
LLJ
10.

E  8

f=  6

I  4

I  2

   0
  TOTAL OIL
 0	i	
                          r2-0.31
	-?		_t__
   MONO AND DICYCLIC AROMATICS
    0      20      40      60      80     100     120
                          TIME OF DEPURATION, days
                                        140     160     180
EXHIBIT  6:  CONTENT OF TOTAL OIL AND AROMATIC HYDROCARBONS
   IN SEDIMENT AS A FUNCTION OF DEPURATION  TIME
    16 C
  o	o DOCOSANE
  D	D NAPHTHALENE
  A	A PHENANTHRENE
                                                     PHENANTHRENE
     40
                                 120       160
                            TIME OF DEPURATION (DAYS)
                                                  200
                                                           240
  EXHIBIT 7:   CONCENTRATIONS OF DOCOSANE,  NAPHTHALENE,
     AND  PHENANTHRENE  IN SEDIMENT  AS A FUNCTION OF
     DEPURATION TIME
                           119

-------
(DMN) toward the elimination of N and later MM.  This relationship con-
cerning greater retention of the less soluble and probably less degrad-
able dimethyl-isomers has been observed while animals are depurating in
clean water, but not during constant long-term exposure  (Anderson,
1977).   While there are some small fluctuations in the levels of naph-
thalenes in control animals, the background concentrations resulting
from absorbence of biogenic compounds in the 221-228 ym wavelength range
(Neff and Anderson, 1975) are quite small.

     The growth rates for these benthic arnphipods are illustrated in
Exhibits 8b and 8c.  While the maximum length and weight levels attained
by all three groups of animals were approximately equal, the control
amphipods reached this size at least 3 weeks sooner.  This indicates
that, during early stages of the exposure, growth was significantly sup-
pressed.  Such an effect could either be the result of smaller individ-
uals being more sensitive, or acclimation to the pollution stress taking
place after a period of time.  In any case, the rate of growth was def-
initely reduced at the 7-week interval.

     Field Exposure to Oiled Sediments

     Sipunculids and Macoma placed in oil-contaminated sediment and
transferred to the field for 40 days contained significant amounts of
PHC  (see Exhibit 9).  This is in sharp contrast to the tissue levels of
the  filter-feeding clam, Protothaca staminea.  From these results, it
is clear that the two deposit-feeders obtained the hydrocarbons from
ingestion of sediment and not from the water column.  The distribution
of specific hydrocarbons within Macoma and Phascolosoma is quite sim-
ilar.  Triaromatic compounds (e.g., phenanthrene) and n-alkanes were
found at the highest concentrations.  These data give additional evi-
dence of the significance of naphthalenes in organisms exposed to whole
oil  or extracts, and the di- and trimethyl-isomers were, as usual, pres-
ent  at higher concentrations.  It is interesting to note that the parent
compound  (naphthalene) was absent from the tissues, as shown in earlier
research  (Anderson, 1977) and the amphipod experiments.

     The final aspect of the biological research relates closely to the
analyses of field sediments discussed above.  Substrate collections
have been taken at four time intervals, and there are four sets (six
each) of sediment boxes in the intertidal zone.  At this time, the sam-
ples of the first two collections have been sorted for biota, but only
a partial list of organisms and data on the numbers of the two most
prominent bivalves have been prepared.  The incomplete list of species
found thus far in the sediment samples is shown in Exhibit 10.  As in
similar benthic studies, three classes of invertebrates are well repre-
sented (Polychaeta, Mollusca, and Crustacea).  A more complete picture
of the number of species and individuals present in clean and contami-
nated sediment will be available by summer.
                                120

-------
EXHIBIT 8:  EFFECTS OF CONSTANT-FLOWING PRUDHOE BAY CRUDE
  OIL EXTRACT ON THE AMPHIPOD, ANONYX (Vertical bars rep-
  resent standard deviations, where the initial sample was
  20 animals and others consisted of 10 animals each, with
  2 animals per naphthalenes analysis.  Tanks 4 and 5 were
  exposed, while Tank 1 is a control.  Exhibit 8a is up-
  take of naphthalenes; 8b is growth in length; and 8c is
  growth in weight.)
                        121

-------
EXHIBIT 9:  PETROLEUM HYDROCARBON CONTENT (yg/g wet; ppm)  OF SIPUNCULID WORMS (PHASCOLOSOMA)
AND CLAMS (MACOMA AND PROTOTHACA)
'EXPOSED IN THE FIELD TO
OILED SEDIMENT FOR 40 DAYS
(Number in parentheses represents the number of specimens in each sample)
Hydrocarbon Content
n-alkanes C)2-C29
1 and 2-methyl naphthalene
total dimethyl naphthalenes
total trimethylnaphthalenes
total triaromatics
total aromatics
Oil-exposed
Sipunculids
(3)
1.90
0.23
0.60
0.95
2.25
4.03
Oil-exposed
Sipunculids
(2)
0.73
<0.01
0.15
0.44
0.77
1.36
Control
Sipunculids
(3)
<0.10
<0.01
<0.01
<0.01
<0.10
<0.15
Oil -exposed
Macoma
(3)
0.69
0.06
0.89
0.90
1.90
3.75
Control
Macoma
(2)
<0.10
<0.01
<0.01
<0.01
<0.10
<0.15
Oil-exposed
Protothaca
(2)
<0.10
<0.01
<0.01
<0.01
<0.10
<0.15

-------
            o0:   LIST OF ORGANISMS  COLLECTED
          FROM  FIELD  RECRUITMENT SEDIMENT  BOXES
                               Polychaeta

                        Platyne.ve.is bioana.li.av.la.ta
                        Armandia bioculata
                        Lepidonotus caelorus
                                Mol1usca
       (Bivalvia)
Clinocardium mittailii
Prototka.ca staminsa
Macoina sp.
Coopevella. subdiapfazna.
Mytilus edulis
Mysella tumida
 (Gastropoda)
Alabina sp.
Margarites sp.
Lacuna sp.
Odostomia
Bittiwn
                                 Crustacea
                                    ampl'icauda
                       Gnorimosphzeroma Ofegonensi
                       Pagurus grano six-arms
                       Virmixa sp,
                       Hsrnigvapsus  sp.
                       Cvjnello, sp.
                       Balanus sp.
                       Leptoahelia  sp.
                       Corop'hiwn sp.
                                 123

-------
     The two most prominent species were the  bivalves (Mollusca)
Cooperella subdiaphana and Mysella tumida  (see  Exhibit 11).  These
species were found  at both time intervals  and in every sediment sample.
Exhibit 11 not  only illustrates the number of individuals of these two
species, but also the ratios of Cooperella to the total and the level
of sediment hydrocarbon contamination.
     EXHIBIT  11:   RELATIONSHIP BETWEEN THE HYDROCARBON CONTENT OF
               SEDIMENT AND THE DISTRIBUTION OF  BIVALVES
                          FROM THE SEDIMENT
            NUMBER OF BIVALVES
       Cooperella   % C.s.
       subdiapham of total
Mysella
timida
Control or
 Exposed
                                                       PPM
        Total*
Docosane  Saturates
           Total*
Phenanthrene Aromatics
  0

  65



 114V



 129



 220**
61
36
34
127
47
170
57
54
33
17.0
18.6
15.5
17.8
15.7
43.4
18.5
18.3
14.0
298
157
185
588
253
222
251
241
203
                  16     275
                         34
                   0.5    8.6
                                           0.3     5.1
                                  0.4
                                  0.06
                                                          0.08
                                          140
                                           18
                                            2.8
                                            3.7
*    Except  for zero time data, these are  estimates  based on percentages
in initial samples.

t    These data are  from a second field installation,  which has not yet
been analyzed for hydrocarbon content.  The  clam populations are calcu-
lated  from replicate subsamples.
**
     Biological data are not yet available.
     As discussed in the analytical section,  the  amounts of saturate
and aromatic  hydrocarbons decreased considerably  by the 65th day in the
field.  It  should be noted that the mesh bottoms  on the boxes allowed
natural percolation of sea water to occur  as  the  tide level fluctuated.
This process  no doubt served to keep the sediment aerobic and probably
increased the rates of hydrocarbon leaching and degradation.  Some of
the smaller organisms may have entered the boxes  via the 1.6 mm open-
ings in the mesh.
                                 124

-------
     It is obvious that the sorbed hydrocarbons did not restrict re-
cruitment by the two  clam  species.  The numbers of individuals in oiled
sediments are, in most cases, very similar to those in control boxes.
ine two sets of data  points on day 114 deviate from all other findings.
The high number of both species in control sediment is somewhat sur-
prising, but the proportion of Cooperella to the total indicates the
data are consistent with the general pattern.  More difficult to ex-
plain is the sharp increase in relative proportion of Cooperella  (43%)
in exposed sediment,  as compared with all other collections.  The number
of Mysella in  this sample  is very close to all other collections, so it
would appear the the  abundance of Cooperella in this oiled sediment may
be because of  some chemical attraction and resistance.  Similar associ-
ations of Cooperella  with  oil-contaminated sediment have been observed
earlier by Vanderhorst  (unpublished data).  Future biological and chem-
ical analyses  of later samples will aid in the interpretation of these
findings.
DISCUSSION

     The  methods  developed to characterize  the hydrocarbon components in
the  flowing sea water exposure system have  been shown to be sensitive
and  accurate.   In addition,  the analytical  results demonstrated that the
delivery  system produces  a consistent level of contamination in the
exposure  tanks.   When the methods  used to analyze Prudhoe Bay crude oil
for  component hydrocarbons (gas capillary chromatography) were applied
to both fine and  coarse sediments,  the separation of specific compounds
was  extremely good.

     On the basis of the  results generated  thus far, the experiments
have been useful  in  demonstrating  the differences in the potential per-
sistence  of crude oil utilizing undisturbed fine-textured marine sedi-
ments  under anaerobic laboratory conditions compared to a field experi-
ment using  coarse beach sediment under aerobic conditions.  Furthermore,
these  experiments have provided sets of samples for development of ana-
lytical techniques appropriate not only for use in conjunction with re-
lated  biological  studies  of oil-contaminated  sediments, but also to
studies involving the uptake and metabolism of oil by marine organisms.
To cite an  example,  we are currently developing HPLC techniques to be
used alone  and  with  our existing capillary  chromatography methods to
replace conventional column methods.   These analytical techniques,
coupled with the  appropriate biological experiments, will provide much
needed information on the chemical,  physiological, and community effects
that would  be observed for marine  organisms exposed to these types of
experimental conditions.

     It was found that the flowing exposure system delivered from 0.22
to 0.24 ppm of  total hydrocarbons  (IR), and that level slowed the growth
of Anonyx during  the first 7 weeks of- exposure.  Anderson and Kiessler
have" compiled  an  extensive amount  of unpublished data on this species,
and  some  aspects  of  these findings can be used for comparisons.


                                 125

-------
In static bioassays using sea water extracts of Prudhoe Bay crude, an
initial concentration of about 6 ppm is required to produce the 96-hr
LCt^Q.  Animals dying during exposure contain from 5 to 11 ppm total
naphthalenes, while those surviving at lower concentrations contain
from 0.9 to 1.8 ppm total naphthalenes.  The tissue concentration of
amphipods exposed for 14 weeks at very low concentration  (0.23 ppm) was
1.8 ppm total naphthalenes, which indicates that our exposure is simu-
lating chronic contamination which is very near the threshold of toler-
ance.  Further evidence of this is the marked reduction in production of
viable offspring by gravid females, which we observed in mid-February
1977.  In other words, long-term exposure to a concentration which is
0.04 times the static system 96-hr LC5Q value has reduced growth of
adults during the early stages of exposure but not inhibited maximal
growth.  The final effect of this exposure, which is most meaningful,
is the inhibition of reproduction; however, this was not observed until
19 to 20 weeks after initiation of the experiment.

     To our knowledge, there have only been two previous reports of
hydrocarbon uptake from sediment by deposit-feeding marine organisms.
Anderson et al.  (1977) described the rates and extent of naphthalenes
uptake by the sipunculid (Phascolosoma) exposed to sediment that had
either been mixed with Prudhoe Bay crude or had received a layer of oil
on the surface.  These animals did contain contamination after 2 to 14
days of laboratory exposure, but the levels approximated that in the
sediment, and depuration of 6 to 14 days resulted in almost complete
loss of naphthalenes.  Rossi and Anderson  (1977) exposed the polychaete
(Neanthes arenaceodentata)  to sediment containing 9 ppm total naphtha-
lenes for 28 days, and the animals never exhibited levels above back-
ground.  When these worms were fed artificial detritus containing about
10 ppm   C-2-methylnaphthalene, they found the accumulated 14C-activity
was voided in the feces during 24 hr with clean food and water.
Roesijadi et al.  (unpublished) reported on the uptake of naphthalenes
by the detritus-feeder, Macoma inguinata.  The contamination that was
present in these clams apparently came from small amounts released to
the water and not from the ingested detritus.

     This paper, for the first time, presents data on the uptake of
numerous PHCs by three species present in contaminated sediment held
for a long period (40 days) in the natural environment.  Table 9 demon-
strated the extent of hydrocarbon uptake by the two deposit-feeders, in
contrast to the lack of contamination in Protothaca, which is a filter-
feeder.  This is probably the most significant approach to determine
uptake of hydrocarbons via ingested sediment particles, as all three
species were surrounded by oiled substrate.  While n-alkanes were pres-
ent in Macoma and Phascolosoma at significant levels, the aromatics, and
particularly the triaromatics, were found at rather high concentrations.
This approach will be continued to determine the rate and extent of
uptake, as well as the specific compounds of significance.  These pre-
liminary findings demonstrate that benthic deposit-feeders take up large
                                126

-------
molecular weight  aromatic  compounds  from polluted sediment.  These hy-
drocarbons are  the most  harmful  to marine organisms, as well as verte-
brates,  including man.

     The flnal  aspect of this  research  concerns the natural rates of
hydrocarbon depuration from oiled sediment and the effects of these
compounds on  the  recruitment of  benthic invertebrates.  The signifi-
cance^of the  analytical  approaches and  findings have already been noted,
and without them  the  biological  data would be meaningless.  The only
other  investigation regarding  correlations between hydrocarbon content
in sediment and the distribution of  benthic organisms is as yet unpub-
lished (Armstrong et  a.1. ).   For  more than 1 year, these researchers have
recorded the  concentration of  naphthalenes in the sediments near an ef-
fluent of an  oil-separator platform  in  Trinity Bay, Texas.  Along three
transects projecting  away  from the source of contamination, they have
recorded the  numbers  of  individuals  and species in sediment samples and
compared these  to the concentrations of naphthalenes present.  Stations
within 500 ft of  the  platform  had depressed benthic faunas, and there
was a  correlation between  levels of  sediment naphthalenes and reduced
populations of  organisms.   A peak  ("stimulation zone") in both numbers
of species and  individuals was found at between 2,250 and 5,500 ft from
the platform.

     While sufficient time has not been available to complete either
chemical or biological analyses  of all  samples collected thus far, we
have not seen an  effect  on recruitment  of benthic bivalves.  As the
hydrocarbon analyses  show  rather rapid  loss from field sediments, the
levels could  have decreased to the point at which summer larvae were not
inhibited from  settling.   There  are  no  apparent effects on the two clam
species inhabiting  these sediments for  approximately 3 months.  Only
by continuing analyses of  later  collections and installing new contami-
nated  sediments,  can  these questions be answered.

     It should  be noted  that this research is ongoing, and the results
presented in  this paper  should be considered preliminary.  Thus far,
the findings  clearly  indicate  that the  approaches and techniques being
utilized have been productive  in deriving much needed information on
the fate and  effects  of  petroleum hydrocarbons in the marine environ-
ment.   Each aspect of the  research has  and will provide sufficient data
to warrant publication separately.
REFERENCES

American  Petroleum Institute.   1958.  Determination of Volatile and Non-
  Volatile  Oily  Material:   Infrared Spectrometric Method.  API Publica-
  tion No.  733-48,  American Petroleum Institute, Washington, D.C.

Anderson, J.W.   1977.   "Responses  to Sublethal Levels of Petroleum
  Hydrocarbons:   Are They  Sensitive Indicators and Do They Correlate
  with Tissue Contamination?"   In  D. Wolfe, ed., Fate and Effects of


                                127

-------
  Petroleum in Marine Ecosystems and Organisms, Proc. of NOAA Symp.,
  Seattle.  Pergamon Press, New York.   (In press.)

Anderson, J.W., L.J. Moore, J.W. Blaylock, D.L. Woodruff, and S.L.
  Kiesser.  1977.  "Bioavailability of Sediment-Sorbed Naphthalenes
  to the Sipunculid Worm, Phascolosoma agassizii."  In:   D. Wolfe, ed.,
  Fate and Effects of Petroleum in Marine Ecosystems and Organisms,
  Proc. of NOAA Symp., Seattle. Pergamon Press, New York.   (In press.)

Anderson, J.W. and S.L. Kiesser.  "Toxicity of a Water-Soluble Extract
  of Prudhoe Bay Crude Oil for the Amphipod, Anonyx nugax, and the
  Uptake of Naphthalenes from Water and Sediment."   (Unpublished manu-
  script. )

Armstrong, H.W., K. Fucik, J.W. Anderson, and J.M. Neff.  "Effects of
  Oilfield Brine Effluent on Benthic Organisms in Trinity Bay, Texas."
  (Submitted to Estuarine and Coastal Marine Science.)

Bean, R.M. and J.W. Blaylock.  1977.  "Characterization of Volatile
  Hydrocarbons in Flowing Seawater Suspensions of Number 2 Fuel Oil.
  In:  D. Wolfe, ed., Fate and Effects of Petroleum in Marine Ecosystems
  and Organisms, Proc. of NOAA Symp., Seattle.  Pergamon Press, New
  York.   (In press.)

Neff, J.M. and J.W. Anderson.  1975.  "Ultraviolet Spectrophotometric
  Method for the Determination of Naphthalene and Alkylnaphthalenes in
  the Tissues of Oil-Contaminated Marine Animals."  Bull. Environ.
  Contain. Toxicol. 14;  122-128.

MacLeod, W.D., D.W. Brown, E.G. Jenkins, L.S. Ramos, and V.D. Henry.
  1976.  A Pilot Study on the Design of a Petroleum Hydrocarbon Base-
  line Investigation for Northern Puget Sound and Strait of Juan de
  Fuca.  Completion Report submitted to Puget Sound Energy-Related
  Research Project, Marine Ecosystem Analysis Program, Environmental
  Research Laboratory, Seattle, Washington.

Rice, S.D., J.W. Short, and J.F. Karinen.  1977.  "Toxicity of Cook
  Inlet Crude Oil and No. 2 Fuel Oil to Several Alaskan Marine Fishes
  and Invertebrates."  In;  Proceedings of Symposium on Sources, Effects
  & Sinks of Hydrocarbons in the Aquatic Environment  (AIBS), Aug. 9-11,
  1976, Washington, D.C.   (In press.)

Roesijadi, G., D.L. Woodruff, and J.W. Anderson.  "Bioavailability of
  Naphthalenes from Marine Sediments Artificially Contaminated with
  Prudhoe Bay Crude Oil."  (Submitted to Environ. Pollut.)

Rossi, S.S. and J.W. Anderson.  1977.  "Bioavailability of Petroleum
  Hydrocarbons from Water, Sediments, and Detritus to the Marine
  Annelid, Neanthes arenaceodentata."  In:  Proceedings of 1977 Oil
  Spill Conference, (EPA, API, USCG), New Orleans.  American Petroleum
  Institute, Washington, D.C.   (In press.)
                                128

-------
Vanderhorst, J.R.,  C.I.  Gibson,  L.J.  Moor.e,  and  P.  Wilkinson.   1976a.
   Continuous-flow  Apparatus  for Use  in  Petroleum Bioassay," Bull.
  Environ. Contain.  Toxicol . 3.7  (in press.)
Vanderhorst, J.R.,  R.M.  Bean/  L-J>  MoorSf  p_ wilkinsonf  c>1.  Gibson,
  and J.W. Blaylock.   I976b.  "Effects  of a Continuous  Low-Level  No.  2
  Fuel Dispersion on  Laboratory-Held Intertidal  Colonies."  .In:
  Proceedings  of 1977 Oil  Spill  Conference,  (EPA,  API, USCG) ,  New
  Orleans.  American  Petroleum Institute,  Washington,  D.C.   (In  press.)

Warner , ^ J . s .   1976.   Determination  of  Aliphatic  and  Aromatic  Hydrocar-
  bons in  Marine Organisms, Analyt. Chem.  48:  578-583.

Wolfe, D.A., ed.   1977.  Fate and Effects  of Petroleum in Marine Eco-
  systems  and  Organisms,  Proc.  of  NOAA Symp. , Seattle.   Pergamon Press,
  New  York.   (in press.)
DISCUSSION

     Dr. Spies;  with respect to the movement of sediment that might
have occurred, could you describe how that was broken down?

     Dr. Anderson:  We  still see the dominant contamination in the top
layer.  This material does not move readily.  Since it is not an area
where there is active movement, we recently mixed in glass beads coated
with barium that can be analyzed to ensure that this material is not
being replaced by other clean sediment.

     From the floor;  You show a marked difference between field weath-
ering and laboratory conditions.  How would you prepare a weathered
fraction test?

     Dr. Anderson:  We  have this large treatment facility with a float-
ing layer of oil on top of it.  The oil builds up to the point where it
must be pumped off.  We have considered using that as our weathered oil.
However, it would not be very good since it contains a few other toxi-
cants from our other systems.  A sharp difference in the toxicity of the
water soluble fraction  results just from putting an aerator in a soluble
extract.

     Dr. Dieter:  In your exposure for the clams, you showed about 4 ppm
accumulated.

     Dr. Anderson;  That was the total analyzed.  The oiled sediments,
the clams, for instance, contained about 600 ppm of total hydrocarbons
to simulate a real spill situation.  So that is not a magnification,
by any means.  This is  why I use the word "uptake," rather than "accu-
mulation," because you  can talk about a large accumulation from water.
From sediment, you are  usually trying to discover if there is significant
uptake.  Over time, there was a large loss of this material.


                               129

-------
     Dr. Davies:  Have you considered the fact that aksane and carbon
tetrachloride may not be able to extract some of the materials?

     Dr. Riley:  As far as the carbon tetrachloride extractions are
concerned, we have proved that you can get approximately 97% recovery
of the oil from the sediments.  Currently, we have an ongoing experiment
to verify a similar type of recovery from the hexane extract.  The tech-
niques are essentially the same except for the solvent, and we expect
that the recovery will be about the same.

     Dr. Anderson:  The hexane in tissue extract is not nearly as effi-
cient as, for instance, diethyl ether.  We always tend to test these to
see what kind of efficiency we are getting.  Using a resin column sys-
tem, we have tried to see if we are collecting all the petroleum and
getting the efficiency of extraction.  However, we are working with
relatively high amounts so it is not a problem.  If we get down to very
low levels, it might be a problem.

     Dr. Davies:  In our laboratory, some experiments with pesticides
have shown that some of it is getting bound in the salmon and it is
then extracted.  So, we are doing tracer experiments.

     Dr. Lefcort:  Are you planning any histolog-ical examination of the
field-exposed bivalves?  We are interested in the neoplasia that occurs
in these.

     Dr. Anderson;  We have proposed from time to time to do some work
with the flat fish in the area.  In fact, I know of some similar ongoing
experiments in the NOAA laboratory.  Frankly, our laboratory is not
geared up for histological examinations.

     Dr. Baumgartner:  Do you have any other data on interstitial anal-
yses in the laboratory sediments to give you any indication?

     Dr. Anderson;  It is obvious underneath a film that is perhaps only
h in. thick.  You can see the sharp break between the upper tan zone and
the black anaerobic mud beneath.  There really is no doubt about it.
However, the chemical measurements have not been done.

     Dr. Baumgartner:  Where is the oil sample?  When you take these'
degradation samples, do you mix a vertical column?

     Dr. Anderson:  With that particular mud system, we take just a
scoop and obtain surface plus 4 or 5 cm beneath.  In the field sites,
we are using PBC cores to take samples.

     Dr. Kunhalt:  You mentioned the reproduction of some of your test
animals.  Which species were they and what did you find about egg num-
bers or hatching?
                                130

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     Dr. Anderson:  We do not have all the data yet because they are
still hatching.  We have looked at the brood pouches of one species of
anthropod that was constantly exposed for  19 to 20 weeks  in the system.
In many of these, the eggs were not completely developed, whereas, by
now, most of the controls have hatched out completely.  The number of
offspring seems to be reduced, but we will not have good data on that
for awhile.  The number living of the juveniles that hatch is dramati-
cally different.  We essentially have not seen any living juveniles
from those females, whereas  all the juveniles that hatch from the
controls are living.

     Dr. Kunhalt:  Were these the animals that did not show any differ-
ence in growth until maturation?

     Dr. Anderson:  That  is  right.  The  last growth point, however, was
14 weeks, which was before we saw the brood pouch development of the
females, so we did not  include the  females.  Essentially, that was non-
gravid  growth.

     Dr. Kunhalt:  What size are they?   Could they be analyzed by normal
methods to get accumulation  in the  eggs?

     Dr. Anderson:  Oh,  yes. We have some other ongoing  studies with
shorter term exposures,  and  about  four different ongoing  experiments
with gravid  female  anthropods.
                                 131

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           ENVIRONMENTAL ASSESSMENT OF NORTHERN PUGET SOUND
                     AND THE STRAIT OF JUAN DE FUCA

                              Howard Harris*
     The environmental assessment of Northern Puget Sound and the Strait
of Juan de Fuca is a 5-year study that addresses the potential marine
environmental and ecological consequences of the intensified petroleum
activities anticipated for the next'decade.  It is one of a series of
projects funded by the EPA that will provide the information needed to
identify and cope with the impact of accelerated energy development.
This research is administered by National Oceanographic and Atmospheric
Administration's  (NOAA) MESA (Marine Ecosystems Analysis) Puget Sound
Project Office in Seattle, Washington.
REGIONAL OVERVIEW

     The Puget Sound region  (see Exhibit 1) has never been a busy petro-
leum center, as it has imported and refined only enough crude oil to
supply its local population.  Since most of the crude has been supplied
via Canadian pipeline, there has historically been only minor shipping
of petroleum and petroleum products through the area's waterways.  Re-
cently, however, the amount of tanker traffic has increased, and a
further intensification of petroleum-related activity is anticipated
for the coming decade.  Two main factors account for this increase:
new suppliers and a potentially larger market.

     Canada has announced that it will cut off all shipments of crude
oil to the United States by 1981.  The Federal Energy Administration
(FEA) has determined that oil delivered in the interim should be trans-
ported directly to the Midwest, which means that all Canadian oil to the
Seattle region will cease flowing in 1977.  Not only does this action
require supplying the Northwest via tanker, but it also results occa-
sionally in a surplus of crude oil in western Canada.  This surplus is
shipped out from terminals near Vancouver through the Strait, of Juan de
Fuca.

     The second factor is possible expansion of local petroleum facili-
ties to serve other marketing areas.   Puget Sound is the closest
U.S.  port to the termination of the Alaskan pipeline.  Its natural
*    Puget Sound Project Office, National Oceanographic and Atmos-
pheric Administration, Seattle, Washington 98109.
                                133

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                                                                                  49'00'N
                                                                                  48-00'N
                                                     123* DOW
EXHIBIT  1:   STUDY  AREA

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deep harbors make it ideally suited to serve as  a major transshipment
point for other regions of the United States.  Under  this  scheme,  crude
oil would be shipped into the region via tanker  and either piped out to
the Midwest, refined and shipped to California,  or both.   The  Oceano-
graphic Institute of Washington, in a study for  the state  legislature,
has evaluated several scenarios and has estimated that Puget Sound tank-
er traffic could increase tenfold by the end of  the century.   No pipe-
line construction is yet underway, and plans for this type of  expansion
remain incomplete, although several area refineries presently  plan con-
struction of deepwater ports to accommodate larger vessels.

     Since tanker routes to the existing refineries at Cherry  Point and
Anacortes traverse major fishery and recreational areas of the Strait of
Juan de Fuca and northern Puget Sound, much local publicity has been
given to the threat of oil pollution to marine,  economic,  recreational,
and aesthetic interests.

     The Washington State Legislature has recognized the need  to formu-
late a realistic energy policy and has taken several steps to  prevent
radical departures from the current petroleum transport methods.   One of
these steps was a moratorium on construction of  new marine oil transfer
facilities; another was the Tug Escort Law, which severely limits  the
movement of supertankers in waters east of Dungeness.  (The Tug Escort
Law has been successfully challenged in court as unconstitutional  by the
Atlantic Richfield Company-)

     In 1974, the Washington Energy Policy Council appointed by the gov-
ernor recommended that area refineries should not be expanded  to serve
more than the traditional marketing areas, and that the existing refin-
eries should all be served by a common-use terminal in or west of  Port
Angeles.  This recommendation was endorsed by the then-Governor Evans,
and the Port Angeles common-use terminal is under heavy study  by all
concerned parties.  In January 1977, Governor Evans was replaced by
Governor Ray-  At this time, the question of state energy policy is un-
resolved, but it is being actively addressed by  the new governor and by
the state legislature.

     In response to this situation, a number of  investigations have been
initiated to provide a basis for rational decisions on the management of
Puget Sound.  These include both investigations  focusing upon  the  exist-
ing refinery sites at Cherry Point and Anacortes (and the possible
transfer site at Port Angeles) and laboratory investigations concerned
with effects of low concentrations of petroleum  upon the native biota.

     The State of Washington Department of Ecology (DOE) has initiated
baseline studies in the high-risk region near the existing refineries at
Cherry Point and Anacortes and in the San Juan Islands.  The program was
initiated in 1973 and is funded at least through a second biennium to
June 1977.  The main accomplishments so far include:  a survey of  the
intertidal and shallow subtidal biological resources through a habitat-
specific analysis of community structure; publication of a literature


                                135

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review on the region's biologic resources; and development of an econom-
ic value index of the resources that might be affected by oil pollution.

     Battelle Northwest has been funded by the U.S. Energy Research and
Development Administration  (ERDA) and EPA to investigate the potential
effects of long-tem exposure to petroleum-derived hydrocarbons through
laboratory studies and in situ exposure of selected communities.  Bat-
telle Northwest has also undertaken an extensive analysis of petroleum
contaminants in water, organisms, and sediments near Cherry Point, Ana-
cortes, and Port Angeles.

     The University of Washington is performing a multiyear sampling
program to determine the distribution of aliphatic, aromatic, and sul-
fur-containing molecules in organisms, sediment, and water at stations
in Puget Sound and along the Washington coast.   The sources and food web
transfer of these compounds are under investigation.

     EPA is funding NOAA to investigate the fate and effects of petrole-
um hydrocarbons and selected toxic metals in marine ecosystems and orga-
nisms.  Although this research is not specific to Puget Sound, a part of
the study will be conducted in this area or in Alaska and should con-
tribute directly to the understanding of potential effects in Puget
Sound.
RESEARCH PROGRAM

     The goal of this research program is to determine the principal
regional characteristics that must be known to assess the potential
environmental effects of increased tanker traffic, petroleum transfer
operations, and refinery capacity within northern Puget Sound and the
Strait of Juan de Fuca.  It is anticipated that 5 years will be required
to complete this program.  Additional research, not encompassed by this
program  (e.g., effects of hydrocarbons upon the biota), must be per-
formed by others before the total implications of increased concentra-
tions of petroleum within the environment can be assessed.

     High priority is given to documentation of the present status of
the marine ecosystem, which presumably is little contaminated by petro-
leum or other substances.  Information needed to deal with a massive oil
spill is also given high research priority, since resultant ecological
effects would be immediately and widely present.  Factors related to
chronic emissions are given a lower priority.  The scope and depth of
research are determined in part by the constraints of available funding
and resources, as well as by the need to perform certain tasks before
the movement of Alaskan oil into Puget Sound in 1977.

     The research is directed toward four major objectives:

1.   Characterize the major marine biological populations subject to pol-
     lution impacts resulting from petroleum transportation and refining


                                136

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2.   Determine the existing distribution and concentration of pollutants
     within the ecosystem that are associated with refinery effluent and
     petroleum

3.   Characterize the principal processes and major pathways by which
     petroleum moves through the marine ecosystem

4.   Provide decision makers with environmental and ecological informa-
     tion and predictions of the effects of oil-related activities upon
     the ecosystem.

     A series of tasks leading to the accomplishment of these objectives
have been defined in the project development plan for this assessment
dated September 1975.  The output from each task will furnish necessary,
and otherwise unavailable, information.  These tasks are coordinated
with other research to avoid duplication of effort.

     The general approach is as follows:

    The biological populations to be investigated include the intertid-
     al and deep benthic invertebrates, nearshore fishes, marine mam-
     mals, and marine and shoreline birds.  These populations will be
     characterized according to their composition, relative abundance of
     species, trophic relationships, and seasonal and spatial distribu-
     tion.

    The present concentrations of selected hydrocarbons and other con-
     stituents of petroleum will be determined in water, sediment, and
     representative animals.  The majority of samples will be collected
     from the intertidal region, but some attention will be given to
     midchannel sampling.

    The major water mass circulation patterns will be determined with
     emphasis upon the surface layer.  The contributions of tides, estu-
     arine flow, and winds will be evaluated.  More detailed, localized
     investigations will be undertaken near existing or potential marine
     terminal sites.  Data will be incorporated into a numerical model
     developed to predict the movement of spilled oil.  Other investiga-
     tions will evaluate the capability of the microbial populations to
     degrade oil and the capacity of naturally occurring suspended mate-
     rials to scavenge hydrocarbons from the water column.

    Available knowledge in selected topical areas will be summarized
     and made immediately available to local decision makers in appro-
     priate formats.

     Ultimately, the results of this research will be summarized and in-
terrelated with data from other petroleum studies to characterize the
major components and processes of the region's ecosystem,  the transport
and fate of spilled petroleum, and the biologic response to oil
pollution.
                                137

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     The principal planned users of the study results are planners and
decision makers at all government levels concerned with the changing
nature of the petroleum industry in Puget Sound, including:

    A change in crude oil supply from overland pipeline to tanker

    A possible expansion of the traditional marketing area and quanti-
     ties handled to include transshipment of crude oil to the Midwest
     or refined products to California

    Construction of a common-use terminal at Port Angeles or Burrows
     Bay to handle supertankers in lieu of expansion at the existing
     refineries

    Modifications and/or expansion of the contingency plan to accommo-
     date the changing situation.

Decisions on these and other questions require detailed knowledge of the
marine ecosystem; this knowlege is largely lacking at present.

     Additional users include the petroleum industry and coastal zone
managers in general.
FIRST-YEAR RESULTS

     Since this research represented a new and major undertaking, a
project development plan was first prepared and approved.   This plan
details the thrust of the research program and the specific tasks to be
accomplished.  In late 1975, arrangements were made with a number of
principal investigators, and field work began early in 1976.  At the
present time, results from the first year are becoming available on the
biological, petroleum baseline, and physical oceanographic program and
model predictions.

     Biological Program

     The biological program has focused initially on three communities
particularly susceptible to damage from floating oil:  the intertidal
biota, the nearshore fishes, and the plankton.  Geographically, the
field programs have concentrated on the Strait of Juan de Fuca, since
information on this area is needed but lacking and since complementary
information on the Cherry Point-Anacortes-San Juan Island region was
being collected by contractors for DOE.  The DOE data have been obtained
and entered into a common data file with the results of this research.

     Plankton investigations have emphasized ichthyoplankton and zoo-
plankton, with less attention to phytoplankton.  Samples have been
collected a total of seven times during 1976, about every 6 weeks
except during midwinter.  A total of nine stations is occupied:  three
across the mouth of the Strait, three in mid-Strait, and three near


                                138

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Port Angeles.  A report on the 1976 work has been prepared by the prin-
cipal investigators.  There is evidence of distinct spatial and temporal
trends in community composition and relative abundance, reflecting both
the oceanic  influence and that of Puget Sound.

     The distribution and abundance  of intertidal  and shallow  subtidal
organisms are being determined at 10 sites along the  Strait of Juan de
Fuca.  A stratified random sampling scheme is being used with quarterly
sampling frequency.  Analyses of the spring and summer samples are
essentially  complete.  Based upon preliminary analyses, the biota is
extremely rich and varied, compared with the San Juan Islands and Puget
Sound.  Many species that were previously undescribed have been found.
Community characteristics vary greatly with substrate type and season.

     At six  of the intertidal stations, nearshore fishes are collected
with a beach seine and a tow net.t  Captured fish are  identified,
counted, weighed, and their stomach contents analyzed to determine pred-
ator-prey relationships.  Preliminary  analysis of the first three col-
lections has disclosed spatial and temporal patterns  in predator-prey
relationships and community composition.  Prey communities found in the
fish stomachs have not yet been compared with those of the intertidal-
shallow subtidal benthic communities.

     Petroleum Baseline Program

     The Puget Sound ecosystem has not been subjected to exposure to
massive amounts of petroleum.  Concentrations of hydrocarbons are pre-
sumed to be  very low, although data to substantiate this contention do
not exist.   Baseline levels will be determined throughout the region,
both in close proximity to existing petroleum transfer facilities and
at remote locations.

     During  1976, preparations were made to conduct the baseline survey.
Funds otherwise provided by EPA were used to establish the NOAA National
Analytical Facility at the Northwest and Alaska Fisheries Center  (NWAFC)
in Seattle.  Concurrently, this program supported the NWAFC in the
development  of specifications for the  baseline study.  This involved two
workshops attended by about two dozen  experts in various disciplines;
identification and testing of various  field and laboratory procedures;
and preparation of a summary report.   Determination of baseline concen-
trations is  beginning in February 1977 at 19 sites.   At each site,
duplicate composite samples of mussels and of intertidal sediment will
be collected on a quarterly basis.  Analyses will include total ali-
phatics, each n-alkane from C,, to C   , total aromatics, and 17 speci-
fied aromatics.

     Physical Oceanographic Program

     The oceanographic program began in February-April 1976 with the
deployment of current meter arrays in  the Strait of Juan de Fuca near
Port Angeles and wind recorders at strategic locations throughout the
                                139

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region.  Three subsurface arrays with four Aanderaa meters each were
deployed across the channel, one at the midpoint and one near each
shore.  An air-sea interaction buoy with current meters was also
deployed near the central mooring to investigate in detail the oceano-
graphic response to winds.

     With the arrival of additional equipment, a more ambitious deploy-
ment was undertaken during the November-February period to investigate
the ocean/wind processes typical of winter conditions.  Five subsurface
current meter arrays were deployed in a cross-Strait transect off Pillar
Point and one in midchannel off Cape Flattery.  Three air-sea interac-
tion buoys were also deployed:  one in mid-channel off Cape Flattery,
one off Pillar Point (in mid-Strait), and one off Tongue Point (west of
Port Angeles).  In addition, five special wind meters were deployed
around the Strait to augment the permanent stations, and six wind
recorders were deployed around the Olympic Peninsula to measure the
larger scale wind patterns.

     Data from the February-April deployment have been processed, and
analysis is underway.  Processing is beginning on the records from the
November-February deployment.

     In another study,  the seasonal and spatial occurrence of suspended
solids is being determined as well as their sorptive characteristics
relative to selected petroleum constituents.  Thirty-three stations in
the eastern Strait and northern Puget Sound were occupied in November
for the purpose of making nephelometer profiles, collecting filter
samples, and collecting 4 g size samples for later laboratory
analysis.

     Information Dissemination and Model Predictions

     The principal vehicle for dissemination will be a series of inter-
pretive monographs on various aspects of the Puget Sound ecosystem/
environment.  During 1976, the specifications for the series were pre-
pared, based upon examination -of comparable series for other geographi-
cal areas, local workshops, and responses to a questionnaire.   There
appears to be a large demand for a series with a level of descriptive
writing about equal to Scientific American with emphasis on graphical
presentations.

     The 1971 annotated bibliography of the Puget Sound marine environ-
ment is being updated and will be available in March 1977.   Content is
being expanded to include a more elaborate indexing system, entries for
the lower Strait of Georgia, and coverage of coastal zone management and
related topics.

     A report has been prepared on the petroleum industry in Puget
Sound.  Included is information on crude inputs according to types,
amounts and sources, refinery processes and wastewater characteristics
                                140

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and product outputs.  Copyright releases are being  sought,  and' the
report should be available this spring.

     Mathematical model development began in 1976 with the  adaptation of
a general oil spill trajectory model developed for  the Bureau  of Land
Management in Alaska to the Puget Sound region.  Work is proceeding on
various required submodels, e.g., a meteorological  submodel to give sur-
face winds, a tidal current submodel, and spreading submodel.   Results
of field investigations are being used to develop and/or verify model
predictions.

     A study was completed in 1976 to assist in defining a  strategy for
possible development of additional models.  The study described the
potential role of models in providing solutions to  questions by state
and local governmental management about marine environmental impacts
that could result from alternative petroleum developments.  Both liter-
ature review and personal interviews with officials from state  and local
governments were employed.  A list of management questions and  impacts
was developed, and the types of models that would be appropriate to the
solution of these questions were identified.
FUTURE DIRECTIONS

     This assessment is now in the second year of a 5-year research pro-
gram.  At the end of this period, the major components of the ecosystem
will have been described and quantified.  Other related investigations
in Puget Sound are on a similar schedule.  The vast majority of this
research is being funded by the federal government.

     It is apparent that an opportunity will exist beginning about 1980
to synthesize the output from these investigations and to deal in a more
sophisticated manner with the consequences of energy development.  Per-
haps it is as much an obligation as an opportunity.  A large investment
of public funds will have been made in this research, and some atten-
tion must be given to ensuring that commensurate benefits are obtained.

     Long-term priorities and requirements in marine research relevant
to energy development are topics to be consided later in this meeting.
These same topics regarding petroleum and Puget Sound are also being
addressed within NOAA, as plans for the MESA Puget Sound Project are
being developed for the early 1980s.  A continuing opportunity for coop-
erative research exists and should be pursued at the appropriate time.
                                141

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DISCUSSION

     Dr. McErlean:  I was curious to know whether you are involved in
any of the 208 planning that area.

     Dr. Harris:  No, we are not.  The 208 planning in that area, as I
understand it, has largely been on the land side, and without too much
consideration given to the marine aspects.

     Dr. McErlean;  Is it just being approached from the land point of
view?

     Dr. Harris:  I believe, at this point, that 208 and coastal zone
management are proceeding rather independently.  The State of Washington
has been involved in coastal zone management or shorelines management,
as they call it, for a number of years.  In fact, their plans have
existed for almost 3 years, although they were just recently approved
at the federal level.

     Dr. McErlean:  It might seem that the data you have might be of
benefit to the 208 agencies.

     Dr. Harris:  Of course, the data are available.  However, I am only
aware of the 208 planning activities in one-half of the water body off
of Seattle.  They are not in the area of the Strait of Juan de Puca,
where we are working.

     Dr. Morgan:  Is there any reason that the biological or hydrograph-
ical surveys have not considered the movements of the deep sediments in
the Strait of Juan de Fuca?  You may be looking at plankton, but not the
deep sediments.

     Dr. Harris:  Later on, we will do some work with the deeper benthic
out in the Strait of Juan de Fuca.  To digress, the Bureau of Land Man-
agement recently held a conference in Portland to discuss the potential
lease sites off the coasts of Oregon and Washington, and one session
dealt with Puget Sound.  The participants were very concerned about the
deeper parts of the Strait of Juan de Fuca.  They felt that offshore
development could well have its greatest impact on the strait.  I am
not sure that we can address that adequately within this program.
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   ENVIRONMENTAL ASSESSMENT OF AN OFFSHORE OIL FIELD: A PROGRESS REPORT

                     Richard J. Berry and Ernest D. McRae*
     Federal research related to offshore petroleum development has
expanded rapidly in recent years because of the legal requirement to
consider possible environmental consequences.  As part of this effort,
the National Oceanographic and Atmospheric Administration (NOAA) ini-
tiated a 5-year assessment of an active oil and gas field through its
component agency, the National Marine Fisheries Service  (NMFS).  In
1975, responsibility for management and implementation of appropriate
studies was assigned to the NMFS laboratory in Galveston, Texas.

     The area selected for study is the Buccaneer Oil Field located
about 53 km (32 miles) south of Galveston (see Exhibit 1).  This field
was chosen because it is isolated from other production areas and has
been in operation long enough to allow development of climax communi-
ties.  Situated in commercial shrimping grounds, it is a focal point for
recreational fishing and diving activities.   Objectives of the study are
to compare ecosystems in the vicinity of a producing field with those in
nearby unaltered areas and to identify changes attributable to pollut-
ants and the presence of structures.
DEVELOPMENT OF THE BUCCANEER FIELD

     Seismic surveys in the Buccaneer Field area were conducted from
1953 through 1959, and Shell Oil Company acquired some 43,000 acres
during the federal lease sale in February 1960.  An exploratory well was
drilled in November 1960 to a depth of 16,088 ft, but it was subse-
quently abandoned because of mechanical problems.  Additional seismic
surveys were performed during 1961-1962 to assess the size, shape, and
complexity of the structural trap.  A second well was drilled in October
1963, and 21 additional wells were drilled within the year.  The "A"
production platform was erected in September 1964, and 15 wells were
drilled from it during the period from January to September 1965.  Plat-
form "B" was installed in May 1965, and 15 wells were drilled from it
*    National Marine Fisheries Service, Southeast Fisheries Center,
Galveston Laboratory, Galveston, Texas  77550.
                                143

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      >Vi)f GALVESTON


        STUDY AREA
f     29-

 GULF OF MEXICO

      94'
                    0= PLATFORM
                    A=WELL JACKET
         EXHIBIT  1:  LOCATION OF  THE  BUCCANEER FIELD STUDY
           AREA AND  STRUCTURES  WITHIN THE FIELD (A and B
           indicate  production  platforms, and X and Y desig-
           nate central  areas to  be compared with the field.)
between October  1965  and July  1966,  More  wells  were drilled between
1966 and 1971, and additional  acreage  was  acquired in the May 1968 lease
sale.  Through June 1971,  81 wells were  drilled,  56 of which were com-
pleted and the remainder plugged.  Currently,  there are 34 active wells,
including 31 gas and  3 oil completions.  As  the  field developed, an on-
shore receiving  plant was  constructed  near Freeport,  Texas, and connected
to the field  (Platform A)  by a 20 in.  diameter flowline, 43 miles in
length.

     At present, structures in the Buccaneer Field include two produc-
tion platforms (A and B),  two  platforms  attached to A and B that serve
as living quarters, 13 satellite  well  platforms,  flare stacks, and pipe-
lines.  Only 6 of the 13 satellite platforms are currently active.  Sat-
ellite platforms or jackets have  helicopter  pads measuring about 20 ft x
20 ft and are supported by four legs fitted  around well caisons.  Under-
water pipelines  connect the satellite  wells  to a production platform.
Once drilling is completed, no gas or  oil  is stored or discharged at the
satellite structures.
                                144

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     Each of the two production platforms have two decks about  150 ft
square and are supported by legs or piles.  Oil and gas from wells asso-
ciated with the production and satellite platforms flow into high pres-
sure vessels on the lower decks.  These vessels separate the gas from
oil and water.  All gas from the field is compressed at Platform B,
piped to A, and then to the Freeport facility.  The oil and water flow
to pressure-step-down vessels, where the two are separated by gravity.
Oil is piped to storage tanks on Platform A, prior to being sent by
pipeline to Freeport.  Produced water flows to skim tanks, where remain-
ing petroleum products are separated and returned to the oil stream.
The water is discharged overboard at rates of 600 to 900 barrels per
day at each production platform.  About 3 barrels of water are  produced
for every 2 barrels of crude oil and condensates.

     Each quarter's platform includes living facilities, a gas-powered
generator, desalinization plant, sewage treatment plant, and a  helicop-
ter pad.  Materials dumped into the sea are limited to edible food
scraps, treated effluent from the sewage plants, and salt water.  Solid
wastes are transported to shore.  Flare stacks associated with  produc-
tion plants are emergency safety devices, which normally have no dis-
charges.  No major oil spills have been reported from the Buccaneer
Field.  Five minor spills between September 1973 and September  1975
amounted to a total of about 6 barrels.
STUDY PLANS

     A pilot study was conducted during November and December 1975 to
determine the suitability of the Buccaneer Field for the proposed inves-
tigations.  Bottom grab samples were obtained from 34 stations in and
near the field.  Analyses included classification of sediments, enumera-
tion of infauna, and measurements of hydrocarbons and trace metals in
the sediments.  Notable findings were that:

1.   Sediments are generally poorly sorted, fine-grained, silty sands

2.   Benthic populations were essentially similar except at one station
     near Platform A, where they were reduced

3.   Hydrocarbon levels in sediments ranged from 0.006 to 0.530 ppm with
     the exception of the station near Platform A, which had a concen-
     tration of 306 ppm

4,   Greatly elevated levels of zinc  (313 ppm) and lead  (41.5 ppm) were
     noted at the same platform station.

Direct evidence of pollution related to oil production was found only at
the one location and no residual effects of drilling activities were
apparent.
                                145

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     Detailed plans  for the  first year of  field work were  developed from
information gained during  the pilot  study,  and contracts were  awarded in
April  1976.  Elements of the study were divided among  the  NMFS labora-
tories at Galveston  and Panama City, Florida, and  four Texas universi-
ties  (see Exhibit 2).  Responsibility for  data management  was  given to
the NMFS laboratory, Bay St. Louis,  Mississippi.   All  project  data  are
forwarded to the Environmental Data  Service, NOAA,  for permanent  stor-
age.

EXHIBIT  2:  LIST OF  STUDY  ELEMENTS,  PRINCIPAL  INVESTIGATORS, AND  CON-
   TRACTORS  DURING THE FIRST  YEAR OF  RESEARCH AT THE BUCCANEER  FIELD
Study Element
Pilot Study
Data Management
Geology
Benthic Fauna
Demersal Fish
Ichthyoplankton
Pelagic Fish
Rig Fauna
Hydrography
Hydrocarbons
Trace Metals
Total Organics
Modeling
Principal Investigator
Dr.
Mr.
Dr.
Dr.
Mr.
Mr.
Dr.
Dr.
Mr.
Dr.
Dr.
Dr.
Dr.
D.
P.
J.
D.
K.
J.
L.
N.
J.
B.
R.
W.
B.
Harper
C. Cook
Anderson
Harper
N. Baxter
Finucane
Trent
Fotheringham
Martin
Middledi tch
Schwarzer
Behrens
Gallaway
Contractor
Texas ASM University
NMFS, Bay St. Louis
Rice University
Texas A&M University
NMFS, Galveston
NMFS, Panama City
NMFS, Panama City
University of Houston
NMFS, Galveston
University of Houston
Rice University
University of Texas
LGL, Limited
     To simplify discussion of the study elements, they are grouped into
benthic, water column, pollutant, and modeling components.  It should be
emphasized that information developed to date is of a preliminary nature
and represents only 6 to 8 months of field activities.  Few substantive
conclusions can be drawn at this time.
BENTHIC STUDIES
     The basic study plan is to compare conditions and fauna in the oil
field to nearby control areas, which presumably are similar but unaf-
fected by either the structures or pollutants.  Exhibit 1 illustrates
                                 146

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the relative positions of the oil field and two  control  areas  (X and  Y)
located in similar water depths  (17-23 m) and  about  8  km (5 miles)  from
the field.  Benthic stations were established  on lines radiating from
platforms A and B to detect effects attributable to  these  structures  and
at other arbitrary positions throughout the study area.

     Geologic Studies

     Geologic studies included magnetic profiling to locate pipelines, a
sidescan sonar survey to detect  irregularities in the  seafloor  surface,
a seismic survey to delineate near-surface structures, and coring to
obtain sediment samples.  The magnetic survey  proved to  be a practical
means for establishing the present position of pipelines,  except in the
vicinity of platforms where the  concentration  of steel structures pre-
vented interpretation of data.   Many  of the pipelines  had  moved several
meters since they were originally set in place.   Pipelines resting on
the bottom were also visible in  records produced by  sidescan sonar.   The
seismic survey was intended to determine near-surface  geological struc-
tures.  Resolution of signals was adequate for penetrations of  40 to
75 m.  Evidence of subsurface structural relief  between  53 and  55 m
indicates the presence of a salt dome beneath  the oil  field platforms
and an old but inactive fault east of the field.

     Sediment samples were obtained from 94 stations (see  Exhibit 3),
using a large corer  (15 cm in diameter) in areas away  from pipelines  and
a  small,  lightweight corer near  the lines.  X-radiographs  of cores re-
vealed a  little by way of sedimentary structure, because of apparent
mixing by benthic organisms.  The character of sediments is variable  and
suggests high energy bottom currents  in the study area.  The origin of
various sediment types is still  uncertain, but areas of  shell and clay
may represent ancient deposits exposed by bottom currents  or structural
uplift.   Sedimentary data suggest bottom scouring within 100 m  of plat-
forms that may operate to disperse materials deposited during drilling.

     Benthic Fauna Analyses

     Because of the danger of rupturing pipelines with heavy gear,  ben-
thic samples were obtained by divers  pushing Ekman grabs into the sedi-
ment.  Triplicate collections were made at 48  stations in  July  1976,  and
about the same number are to be  obtained during  three  other seasons.
Subsamples were removed from each grab with plastic  tubes  for counts  of
meiofauna.  Station locations correspond to the  sediment stations radi-
ating from platforms A and B.  Counts of macrofauna  from July collec-
tions indicated depressed populations in the immediate vicinity of the
production platforms and possible zones of higher than average  numbers
at distances of 500 to 1,500 m.  Differential  distributions of  various
forms make further interpretations difficult.  Polychaetes are  the dom-
inant form  (50-86% of the total), followed by  lesser numbers of crusta-
ceans and mollusks.  Meiofauna counts also appear to decrease in the
vicinity  of production platforms, but exceptions occur.  Nematodes and
                                 147

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foraminifera comprise larger proportions  of  populations near Platform A
than elsewhere.
                      0   1  Kilometer
                      Q      1 Mile
                     D PLATFORM
                     A=WELL JACKET
                     - SAMPLING STATION
74
V
76
77
94 95. n^.'^J^oi
A89a 'n
83. 86' 34~i.. . 37 9'3 "
82 ? "fi7
B 435 38 "'A 3'9 71
6 A 4'8 > H
100.
78 61 A 62 70
66
67
X
69
        EXHIBIT 3:  LOCATION OF BENTHIC STATIONS  WITHIN THE
          BUCCANEER FIELD  (These stations are  used  for  col-
          lection of sediments and benthic  fauna.)
     Demersal Fish  Collection

     Trawl and trap collections  of bottom fish were completed in May,
August, and November 1976  and  are  scheduled for February 1977.  To
permit comparisons  of species  and  sizes of fish and crustaceans, the
sampling design  includes collections in the two control areas and
within the oil field complex.  The presence of pipelines and diffi-
culties of navigating a large  vessel within the field made it imprac-
tical, however,  to  trawl close to  structures,  where species differences
would be expected.   Traps  are  only moderately  effective in catching
the species of interest.   To date,  57 species  of finfish and 23 species
of crustaceans have been caught, but differences between areas are not
apparent.
                                148

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WATER COLUMN STUDIES

     In this study area, we have analyzed  the presence  of ichthyoplank-
ton, pelagic fish and  structure-related fauna,  and collected hydro-
graphic data.

     Ichthyoplankton

     During the first  year, plankton  studies are  limited to  enumerating
fish eggs and larvae in a further attempt  to distinguish possible  dif-
ferences in fish inhabiting the control and oil field areas.   Collec-
tions are on a quarterly schedule  (May,  August, November,  and February),
and data from the first two cruises have been processed.   Oblique  tows
are made with paired Bongo nets  (61 cm  in  diameter)  that have mesh sizes
of 0.333 mm and 0.505  mm.  Each net is  fitted with a flow meter  to mea-
sure water volumes passing through the  net.  Neuston net tows are  also
made at each station to sample surface  plankton.   The mouth  of the neus-
ton net measures 1.0 x 0.5 m, and the mesh size is 0.505 mm.   This net
provides older larvae  that are not vulnerable to  the Bongo nets.   Sta-
tion locations for plankton collections include four within  the  Bucca-
neer Field, one in each of the two control areas,  one situated 8 km
shoreward of the field, and one an equal distance seaward.   A total of
14,496 fish eggs and 2,439 larvae were  caught, during the May and August
cruises.  Among the larvae, 31 families, 33 genera,  and 19 species were
identified.

     By far, anchovies were the most  abundant group at  both  times.  To-
tal egg numbers were significantly larger  in the  field  than  at the con-
trol stations.  Larval numbers were not clearly different in these
areas, but more families were found in  collections from the  oil  field.

     Pelagic Fish

     Many species of pelagic and reef fish are  attracted to  offshore
structures; consequently, recreational  fishermen  and divers  tend to con-
centrate their efforts near oil rigs.   Observations  of  fishing activi-
ties on randomly selected days during the  week  and on weekends between
June and early November indicated that  98% were operating within the oil
field area.  Of these, approximately  75% were bottom fishing,  20%  were
trolling, and 5% were  diving.  Red snapper constituted  86% of the  bottom
fishermen's catch, while king mackerel  represented 58%  of troller's
catches.  Divers appeared to divide their  efforts among  spadefish,  red
snapper, amberjack, and cobia.  The number of fish speared per hour was
considerably lower than the number taken by hook  and line.

     Attempts to use direct sampling  methods to estimate stocks of the
various pelagic species met with limited success.   Each of the methods
used (i.e., trolling,  gillnetting, and  longlining)  is selective  in terms
of species and size of fish.  Data developed from trolling in a  stan-
dardized manner indicated that king mackerel are  strongly attracted
to platforms.  Amberjack, cobia, and  bluefish were caught only near


                                149

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structures.  Surface and bottom gillnets as well as longlines caught
fish but were damaged by sharks to such an extent that they were imprac-
tical to use.  Currently, quantitative photographic techniques are being
tried as a means of estimating species abundance near platforms.

     Structure-related Fauna

     Organisms attached to the supports of oil rigs represent communi-
ties that obviously would not be present in the absence of structures.
Furthermore, they exist in areas of potential hydrocarbon pollution and
may include useful indicator species.  The composition and production
rates of communities associated with the Buccaneer Field are being stud-
ied at Platform A and the southernmost satellite platform.  Comparisons
to an area without structures are made through counts of meroplankton
and fish in a control area 8 km away from the field.  Grazing rates by
fish are inferred from stomach analyses.  Information developed to date
is primarily taxonomic but includes the observation that barnacles are
numerous to depths of 10-14 m from the surface.

     Effects of oil field structures on the migratory behavior of birds
are being investigated through observations made from Platform A and
from a vessel anchored in one of the control areas.  Sightings during
the spring migration period gave counts of 59 species near the platform
and none in open water areas.  A total of 61 birds that had died of ex-
haustion or starvation were found on the platform.  None of these ap-
peared to have collided with the structure.  Only eight species were
noted from the platform and five in the control area during the summer.
Observations during the fall migration produced counts of 30 species
from the platform, 10 in the control area,  and no dead birds.

     Hydrography

     Hydrographic data are being collected to assist in development of
diffusion models related to produced water discharges and trajectory
models applicable to offshore oil spills.  Routine temperature and sa-
linity measurements are also made to support other investigations.  Pri-
mary emphasis is placed on water flow characteristics as documented by
current meter readings, drift bottles, and transponding buoys.  Between
May and October, 579 drift bottles were released in the oil field and
control areas; of these 130 were recovered and returned.  From July to
October, 25 buoys were released to measure water movements at the sur-
face and at 10 m below the surface.  Positions of the buoys were fixed
daily from shore stations over periods ranging from 1 to 15 days.

     Transponding buoy data are extremely valuable as compared to data
from drift bottles.   Buoys permit analysis of short-term drifts result-
ing from wind or tidal influences and movements away from shore, whereas
drift bottles are returned only if they reach the shore.  To date, in-
formation from the two sources has indicated that shoreward drift compo-
nents exist in many months when predominant flows are offshore or paral-
lel to the beach.
                                150

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POLLUTANT STUDIES

     In this study area, we have analyzed hydrocarbons, trace metals,
and total organic concentration.

     Hydrocarbons

     During the first year, objectives of this activity are to survey
the hydrocarbon content of oil and discharges from production platforms
and of water, sediments, and biota in the oil field and control areas.
The biota include plankton, demersal and pelagic fish, benthos, if
available, and barnacles from both production and satellite struc-
tures.  Analyses are performed by gas chromatography and combined gas
chromatography-mass spectrometry methods.  Techniques are based on
specifications of the Bureau of Land Management in its South Texas
outer continental shelf study.  Quality control samples have been ana-
lyzed by chemists at Texas A&M University and the National Analytical
Laboratory, NMFS, Seattle, Washington.  The high levels of hydrocar-
bons found in sediments near Platform A during the pilot study were
absent during the summer of 1976.  Comprehensive analyses and inter-
pretations of study results are not available at this time.

     Trace Metals
      Investigations of trace metal concentrations include analyses of
all bottom sediment samples, selected suspended sediments, barnacles,
fish, and plankton.  Determinations include barium, copper, chromium,
cadmium, lead, manganese, mercury, strontium, and zinc.  Currently
available information suggests that lead and zinc concentrations in
sediments near the two production platforms are well above background
levels.  Lead ranges from 5.5 to 11.8 ppm throughout the study area,
except within 120 m of Platforms A and B, where values up to 100 ppm
are common.  Likewise, zinc concentrations range above 100 ppm in the
vicinity of the two large structures, while the average value for the
entire area is 41.5 ppm.  Enrichment of chromium, copper, and cadmium
sediments is not apparent.  Analyses of other elements and other sample
materials are not completed.

      Total Organics

                13  12      14
      Ratios of C  /C   and C   dating techniques are being employed to
determine the depth and extent of sediments originating from drilling
operations.  Currently, samples obtained by long and short cores at
stations radiating from the production platforms are being analyzed.
MODELING

     Original study plans called for modeling studies to assist in inte-
grating hydrographic and ecological data.  These efforts were postponed
                                151

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 until the  first year  of  survey  work had progressed to a point where
 requirements could be evaluated,   In September  1976,  a contract was
 awarded  to develop a  conceptual model and to outline  second-year study
 plans that would produce the desired types of data.   In October,  this
 approach was discussed with members  of  a user group to elicit comments.
 The user group  consists of representatives of eight state and federal
 agencies that may  be  expected to make use of information developed  from
 the Buccaneer Field study.  Their  recommendations  helped to  ensure  that
 ensuing  studies will  be responsive to needs of regulatory agencies.

      The conceptual model, which summarizes trophic dependencies within
 the oil  field ecosystem, will serve  several functions  during the second
 year  of  work.   It  is  essentially a framework for managing activities and
 organizing information.  As such,  it should improve the efficiency  of
 field activities and  integration of  results.  When sufficient data  be-
 come  available,  the conceptual model will be converted to a  mathematical
 model to enable hypothesis testing and  predictions of  environmental
 impacts.

      Research plans beyond 1977 will be designed to fill gaps  in infor-
 mation required for the model and  to verify impacts suggested by field
 observations.
DISCUSSION

     Dr. Baumgartner:   You said you could not find any evidence of
drilling muds?

     Dr. McRae;  There is a definite pattern in the organics, as along
the coast, that indicates that the older material is there, but so far,
it is not correlated to the currents or this work.

     Dr. Baumgartner:   But you did say you found some on the basis of
zinc and lead near the platforms.  How close?

     Dr. McRae:  As close as the batteries thrown over the side.  How-
ever, we are working under the platform with the divers, and the pol-
lution can be carried by a current.  Next year, we will examine some of
the fish that have sores.  Quite a few of these reef-type fish that are
associated with the platforms have sores, which may be carcinogenic.

     From the floor:   Do the fishermen, the recreational fishermen,
remark upon the quality of the fish that they catch?

     Dr. McRae:  They notice the sores; but they can cut them out with
a knife, so it doesn't bother them.
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        A STANDARD PROGRAM FOR ENVIRONMENTAL IMPACT ASSESSMENT:
                    PHASE I-ICHTHYOPLANKTON SAMPLING

              Martin A.M. Hyman, Ernesto Lorda, and Saul B. Saila*
     The pressing need for enlightened management and regulation of
estuarine ecosystems is made more critical by the difficulty of pre-
dicting the consequences of man-induced perturbations.   An estuary is a
very complex system with large numbers of biological, physical, and
chemical components.  The physical variables vary with time at a fixed
point in space, mainly as a result of water movement of various scales
according to the principles of dynamic oceanography.  Biological vari-
ables and some chemical variables vary as a result of the same processes,
but also according to principles inherent in biological material.  These
may be independent of physical water movements or may enhance or reduce
their effects.  Interactions among these components are characteris-
tically nonlinear and involve intricate feedback loops.

     An adequate or comprehensive survey of attempts to model ecosystems
or even specific estuarine ecosystems involves much more effort than is
available for the purpose at hand.  Suffice it to say that most pre-
dictive modeling efforts to date have involved viewing ecosystems as
consisting of interconnected compartments (sometimes of several compo-
nent species), and interest is centered on the amount of energy or
material in each compartment and the flow rates between compartments.
The ecological processes going on within the compartment among various
species are assumed to average out into some fairly simple behavior for
the compartment.

     The complexity of estuarine ecosystems and the growing need for
proper management decisions and regulations have placed a high priority
on large-scale integrated models.  One example of a complex simulation
model for Narragansett Bay is the recent work by Nixon and Kremer
(1975).  The practical effectiveness of predictions from a model, such
as the above-mentioned, is limited by the size of the model grid as
well as by the accuracy and precision of the model parameters and state
variables, among other things.  In general, it appears that ecosystem
level models, as currently conceived, are able to predict with reason-
able fidelity only the consequences of very gross types of perturbations
and cannot be used for specific practical decisions.
*    Graduate School of Oceanography, University of Rhode Island,
Kingston, Rhode Island  02881.
                                153

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     Recognizing the limitations of these ecosystem models, environmen
tal impact studies have historically developed a so-called baseline and
a so-called monitoring program.  As currently executed, baseline studies
usually consist of a sort of "snapshot," a short-term picture of an
environment, which is variable in both space and time and with little
indication of the magnitude of these scales of variability.  The moni-
toring program has evolved into a systematic collection of data over
time.  Unfortunately, little critical information on the sampling inter-
val, the minimum desired length of the time series record, or the nature
of sampling variability is available.

     Among the host of economic, political, logistic, and statistical
problems involved in the establishment of a standard program for envir-
onmental impact assessment, the latter two areas have received insuf-
ficient attention, in spite of their potential value.  Relatively little
has yet been done to utilize the principles of experimental design in
sorting out effects of spatial and temporal variability as they affect
responses of organisms.

     In general, it is apparent that the intended use of a particular
data set will determine a particular sampling program, and no arbi-
trarily Srawn system for sampling will be suitable for monitoring.  The
reasons for this include the fact that a sampling program for a partic-
ular experiment involves several factors, among which the sampling
rate, the duration of individual observations, the accuracy and pre-
cision of observations, as well as the accuracy and precision of timing
(synchronization) are important.

     It is suggested that a successful environment impact assessment
program is predicated on a sequence of carefully planned experiments
designed to answer specific questions, rather than on a general measure-
ment system.  This carefully planned standard program can then be used
for establishing suitable guidelines for more routine types of assess-
ment.

     To assign sources of variability in ichthyoplankton surveys more
accurately, an intensive sampling scheme was proposed for a single site.
The purpose of this study was to work toward producing a standard pro-
gram for ichthyoplankton sampling.  If variability is assigned to
various' effects, as opposed to being confined into an overall error
term, data will be of an overall higher consistency and, thus, generally
more useful.  Specifically, once this variability is better understood,
any modeling effort should prove more productive.
COLLECTION OF DATA

     To intensively look at variability within a station, we chose a
single sample site in West Passage of Narragansett Bay.  Because this
was an "average" site, it was felt that its results will be directly
applicable to any sampling program.  While interstation variability is
important and assumed to exist, it was not within the scope of our study.


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     The selected sampling site was  1/4 mile  off  the  southwest tip of
Prudence Island.  This area has a relatively  uniform  depth of approxi-
mately 50 ft and is not located in a high  traffic zone.   The  50 ft
depth permitted 4 min duration double oblique tows to be  taken.   The
chosen site was marked by bearings and fathometer traces  and  buoyed to
ensure geographic repeatability.

     Beginning at 8 a.m. for  24 hours on 24 June,  28  June,  1  July,  and
7 July 1976, sampling was conducted  hourly for 24 hours.   Each hour,
two tows were taken, one with the tide and the other  against  it.   A
total of 335 samples was taken.  Although  problems were encountered on
day 1, virtually complete sample sets were taken  on each  of the subse-
quent three days.  These last three  days comprise our data set.   Conse-
quently, for the purpose of analysis, only 280 samples were taken.   The
initial design called for sampling over a  2-week  period;  however,  the
program did run over into a third week.

     The sampling gear chosen for the study was 60 cm Bongo nets,  both
sides containing 505y mesh.   Because of the large mass of historical
data on this gear, it seemed  an appropriate starting  point in spite of
recent additions to the gear  collection.   It  was  expected that the  two-
net system would produce replicate tows; however,  A-B variability  was
measured.  Two General Oceahics flow meters were  used:  one mounted
inside one of the nets and the other external to  the  pair.  For all
practical purposes, this meter is the existing industry standard.   In
addition, a dual-axis electromagnetic current meter with  deck readout
was incorporated into the gear.

     In the original sampling design, we proposed to  compare  the results
obtained from an electromagnetic  (EM) current meter and General  Oceanics
 (GO) flow meters.  It was initially  felt that the GO  meters were a  ques-
tionable source of volume information.  However,  the  meter comparison
proved impractical.  Although several successful  tows were made  with  the
EM internal to the nets, when it was placed externally, an irreparable
loss of cable continuity resulted in premature termination of the  ex-
periment.  This loss was not  correctable within our sampling  period;
however, the nets fished positively  throughout the initial tows  and did
not backwash at any time.

     As a result of the EM failure,  the GO meters became  the  sole  source
of volume data.  Only volume  data and average speed were  obtainable from
these units; however, the meters did prove to be  remarkably consistent.
Three meters were used and rotated each 8  hours to check  for  meter  bias.
None was determined to exist.  For each tow,  internal and external
readings were taken.  In all  but several cases, the internal  meter
reading was between 10% and 17% higher than the external  meter.  This
information is consistent with the findings of the MARMAP studies  (per-
sonal communication with Robert Marak, National Marine Fisheries
Service).  The consistency of the ratios of internal  to external read-
ings deviated by less than 5% over 168 tows.   We  considered this con-
sistency to be more than acceptable  for the purposes  of our comparisons.


                                155

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     In conjunction with the sampling program, various auxiliary data
were collected.  Ambient light was measured by a standardized light
meter reading taken in a northerly direction 30 over the horizon.
Cloud cover and wind speed were estimated in a standard manner.  A water
temperature profile was taken hourly.  No salinity data were collected.

     The platform used for sampling was the University of Rhode Island's
R.V. Dulcinea, a 39 ft converted U.S. Navy utility boat.  Engine rpm
were maintained constant to eliminate differential organism response,
except as it related to tidal and wind effects.  Gear was lowered to
just above the bottom with the vessel at standstill.  When an appro-
priate length of two wires was let out, the tow began.  Attempts were
made to maintain average vessel speed at 2.5 kt (1.25 m/sec).  This
technique varies from traditional sampling with Bongo nets in that the
vessel is stationary when the gear is deployed; thus, "fishing" does
not begin until desired and the meter readings reflect the tow period
more accurately-  Checks made with the electromagnetic meter showed
that this technique produced a uniform tow velocity.  Based on the
MARMAP program of the National Marine Fisheries Service, a recovery
rate of 0.33 m/sec was used, which resulted in an average net velocity
of 1.6 m/sec.

     All the samples were transported to Marine Research, Inc., for
identification and counting.  To determine subsampling error, 10% of
the samples were counted entirely:  one-third of these were from sparse
samples, one-third from intermediate samples, and one-third from dense
samples.  Fc'.* purposes of this check, Marine Research, Inc. , personnel
estimated density, and 10% of the samples were recounted to evaluate
operator error.  Finally, in addition to organism counts, biomass
estimates were computed for each sample using volume displacement.  The
samples were sealed in preservative for purposes of archiving.

     The samples were analyzed for ichthyoplankton eggs and larvae and
zooplankton and benthic organisms.  Because of its mesh size and de-
sign, the gear was not efficient for zooplankton or benthics; however,
we felt that this lack of efficiency was uniform over the sampling
period.  Thus, it would give some indication of variability in a
relative sense.

     The data collected were separated into two categories:   physical
data and species counts.  The species counts were further separated into
four classes:  ichthyoplankton eggs, ichthyoplankton larvae, and zoo-
plankton and benthic organisms.

     For each sample, the number, hour, tow direction, wind speed, sur-
face temperature, mean water column temperature (weighted),  bottom
temperature, water level function, light intensity, tidal factor, in-
side volume, outside volume, time of first oblique tow, time between
tows, and time of second oblique tow were recorded.  With the exception
of the water level function and tidal factor, each parameter is self-
explanatory.  The water level function and tidal factor were computed


                                156

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from the times and heights for the high and  low tides  for those days.
Times and heights were corrected  for Prudence  Island (Exhibit 1).

     The water level  function is  defined  as

     WL  = f(t) = am    [(sin t) + 1] + min,

where

     WL  = water level

     t   = point in time  expressed in radians  or degrees

     am  =0.5  (tidal amplitude)

     min = WL at low  tide (minimum water  level)

     max = WL at high tide  (maximum water level);

so that

           max - min
     am  = 	  .
     The  tidal  factor  is  formally defined  as  the  instantaneous  rate  of
 change of the water  level in meters per  unit  time.   It  is  the first
 derivative of the  above function, so  that:

     TF   = f' (t) = 	 =  am   cost,
                    dt
 where

     TF   = tidal factor  (or slope of  the water  level function WL)

     Negative values correspond to ebb tide or  southward flow

     Positive values correspond to flood tide or  northward flow

     Zero corresponds  to  slack tide.


 ANALYSIS  OF DATA

     There are  two basic  sources of variability in plankton data:  that
 which occurs naturally and that which is introduced  in  the analysis.  We
 assumed that the samples,  if collected rigorously, are  subject  to  as
 little human-induced variability as possible.   In the counting  and
 identification  stage,  any variability is human  introduced.

     To examine relationships  among species counts,  we  will complete
 correlation matrices.  Then, the species groups that demonstrate the

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greatest affinity in response will be correlated separately with the
physical parameters and tow speeds.  The results of this two-step
correlation analysis will be used in determining the actual sources of
most of the observed variability in the species counts.  Multivariate
analyses of variance will then be conducted to investigate the response
of individual species and/or major species groups to the most relevant
variables.

     The final model used in the above analysis cannot be established
a. priori, since its construction will be based on the results of the cor-
relation analysis.  However, it is expected that the response of several
major species (dependent variables) will be measured for roughly the
following sources of variance (independent variables):
     Main Effects                                   Levels     D-.

     D = direction of tow                            2           1

     H = time of day  (within day variance)          24         23

     M = days  (between day variance)                 3           2

     R = replicate tows  (2 nets each tow)            2           1


     The effect "time of day" will be characterized by physical factors,
such as water temperature, light, and tidal factor, on the basis of the
correlation analysis.  Any nonsignificant variable will be pooled into
the error term.

     To estimate the variability introduced in the counting and identi-
fication stages, the sample counts will be examined for counting error.
For each of the whole counts, comparisons with the intermediate split
counts will be made.   In addition, the recounts will be compared to the
initial counts.
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DISCUSSION

     From the floor;  Did you do your tows both  in  daylight  and  at
night?

     Dr. Hyman;  Yes, we started at 8:00 a.m.  and ended  at 7:00  a.m.
the following morning  24 hrs.

     From the floor:  How much tide flow did you have  in there?

     Dr. Hyman:  Oh, a foot and a half.

     From the floor;  You have more than a foot  and a  half,  don't you?
                    (
     From the floor;  Neap tide, 2h; spring tide, maybe  up to  4  ft.
                                  159

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      EFFECTS OF CHEMICAL USE IN OFFSHORE WELL-DRILLING OPERATIONS
                           Norman L. Richards*
     Recent environmental impact statements on oil and gas lease sales
in the Gulf of Mexico have devoted proportionately little discussion to
the environmental aspects of chemical use in well-drilling operations.
This may reflect the fact that few studies have been done, and most are
acute static bioassays that have little relevance to natural conditions
such as drilling mud discharges  (Sheen Technical Subcommittee, 1976).
Of those acute bioassays in the literature, few have reported adverse
effects of drilling fluids or their components.  Field observations have
revealed a variety of marine life in the vicinity of drilling rigs
(Fisher, 1975).  In addition, it is generally assumed that fluids would
either have only a limited local effect near the plume discharge point
or would rapidly be dispersed and diluted in the field (Sheen Technical
Subcommittee, 1976).

     Research on the effects of drilling muds is further complicated by
the chemical complexity of muds.  Additives contain:  pH control prod-
ucts, bactericides, calcium removers, corrosion inhibitors, deformers,
emulsifiers, filtrate reducers, flocculants, foaming agents, lost cir-
culation materials, lubricants, shale-control inhibitors, surface-active
agents, thinners, dispersants, viscosifiers, and weighting agents
(Wright, 1975).  Analyses of potential effects of chemicals used in
well-drilling operations must take into account the variation in mud
and cutting composition from the type of substrate drilled, well depth,
availability of mud components, temperatures generated, relative cost of
components, operator experience, etc.  (Fisher, 1975).

     It is logical that the "King-Muir" report included the following
gas and oil environmental effects objective:  "Objective A:  Determine
the fate and effects in marine and estuarine ecosystems of pollutants
of oil and gas extraction activities...",  and projected as a national
need:  "...Project 2:  Determine the ecological effects of marine and
estuarine organisms of pollutants from oil and gas extraction."  (Work-
ing Group on Health and Environmental Effects of Energy Use, 1974).  A
rigorous biological assessment of potential environmental effects of
*    U.S. Environmental Protection Agency, Environmental Research
Laboratory, Gulf Breeze, Florida 32611.
                                161

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chemicals used in well-drilling  operations has many technical diffi-
culties and is extremely resource  intensive.   Because of these consi-
derations, a limited program  to  study the effects of drilling mud com-
ponents was undertaken at  the EPA  Environmental Research Laboratory at
Gulf Breeze, Florida.  Exhibit 1 illustrates  some areas in which an
understanding of the effects  of  drilling muds is lacking,  i.e.,

    Estimates of the distance that  rigs must be from coral reefs to
     minimize effects from emissions

    Effects of mud on resident  rig  marine life

    Advisability of aquaculture near platforms

    Recommended distance  from rigs  for seafood harvesting.
            Commercial and
            Recreational Fishing
Drilling Mud Barge
                                              SrS04
                                              CHLORINATED
                                              PHENOLS
                                              OTHER BIOCIDES
                 EXHIBIT  1:   SCHEMATIC DIAGRAM OF THE
                   POSSIBLE FATE  OF  ADDITIVES
METHODS

     Our approach  to an assessment of the effects of chemicals used  in
well-drilling operations is  shown in Exhibit 2,  The experimental design
adheres to a tiered  screening sequence,  which has progressively more
complex and time-consuming tests to be run on progressively fewer coy-
pounds.  Xenobiotics are selected from lists of drilling mud components
in current environmental impact statements, with emphasis on those char-
acterized by heavy usage patterns or having suspect toxicity.
                                162

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                                  OPEN OCEAN    \
                                EFFECTS OF MIXTURES   \


                                BEHAVIORAL EFFECTS    \
                               BIOCHEMICAL MECHANISMS
                               COMMUNITY STRUCTURE
                                 AND FUNCTION
                              FLOW-THROUGH BIOASSAYS
                               ACUTE STATIC BIOASSAYS
                             CHEMICALS SELECTED FOR STUDV
               EXHIBIT  2:   ASSESSMENT OF THE EFFECTS OF
                  CHEMICALS USED IN WELL-DRILLING
     Preliminary  screening of selected compounds is accomplished by
acute static 96 hr  bioassays.  They are conducted with drilling fluid
constituents by using laboratory-reared sheepshead minnows  (Cyprindon
variegatus) and grass shrimp (Palaemonetes pugio).  These range-finding
tests are helpful in selecting toxicant concentrations for  subsequent
flowing sea water bioassays for each compound, component, or  mixture of
interest.

     Flow-through toxicity methods were selected for a second tier of
testing because they more nearly approximate in situ conditions.  In
contrast to static  tests, metabolic products and excreta are  removed,
while oxygenated  sea water and toxicant are continuously supplied.   Data
obtained from  flow-through toxicity tests are generally preferable over
static tests as the more precise measure of toxicity and bioaccumulation.
A few compounds are selected from flow-through experiments  for the next
tier of testing:  the composition and functions of estuarine  communities.

     The effect of  selected xenobiotics on colonization of  planktonic
larvae and microorganisms is analyzed by means of the apparatus devel-
oped at the Gulf  Breeze Laboratory by Hansen  (1974).  Exhibit 3 illus-
trates the apparatus used in these studies.  Sea water,  with its natural
component of plankton and microorganisms ,  is pumped  from the estuary ad-
jacent to the  laboratory into the primary constant head box.   Xenobiot-
ics are continuously metered into water after they are siphoned from the
primary to the secondary constant head box; the control apparatus re-
ceives the same flow of water.  Water then flows from the  secondary con-
stant head box to each of the 10 adjacent aquaria, i.e.,  10 replicates
for each treatment, including controls.  At the end of a  9-week exposure
                                  163

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          Primary Constant Head Box
           Secondary Constant Head Box
                                     '\::-&' 'Stand Pipes(200 ml/min)
    EXHIBIT 3:   APPARATUS USED TO TEST THE EFFECT OF XENOBIOTICS  ON
      COMPOSITION OF ESTUARINE COMMUNITIES
period, microflora, macrofauna,  and meiofauna are sampled.  To determine
the effect of  the  xenobiotic  after each treatment, numbers and species
of'microflora, meiofauna,  and macrofauna are compared in control and
exposed aquaria, and  concentrations of xenobiotics in test water and
sediment are determined.   Samples of water from the constant head boxes
are taken twice  a  week,  and sediment cores from aquaria are taken from
each apparatus at  the end  of  the exposure.
RESULTS

     Two compounds were  selected for further study from those screened
in acute bioassays.   Barium sulfate was chosen because "...barite dis*-
charged with drilling muds  is  a  nontoxic substance," and pentachloro-
phenol was chosen because it is  a "...potentially hazardous" drilling
mud constituent  (Sheen Technical Subcommittee, 1976),

     Static toxicity  tests  with  sodium pentachlorophenate have been con-
ducted with larval stages of three marine/estuarine species.  The spe-
cies, the LC50 values obtained,  and the respective exposure period werej
eastern oyster,  < 100 ppb,  48  hr;  grass shrimp, 649 ppb, 96 hr; and pin-
fish, 38 ppb, 96 hr.  A  similar  study with Dowicide G (79% PGP) and
                                 164

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pinfish larvae resulted in a 96 hr LC5Q of  66 ppb.  A bioconcentration
study with oysters exposed to sodium pentachlorophenate  in  flowing  sea
water for 28 days demonstrated the bioconcentration of 80 times  that
measured in the exposure water.

     Effects of pentachlorophenol on development of estaurine  settling
communities were determined by using the apparatus devised  by  Hansen
 (1974) and described under the methods section.  The macrofauna  results
from a 9-week exposure are shown in Exhibit 4.  Individuals and  species
of the numerically dominant phyla  (Annelida, Arthropoda, and Mollusca)
decreased as concentration increased.  Sensitivity differed among spe-
cies of the same phylum.  For example, the  population of the annelid
Haploscoloplos was reduced by exposure to 96 yg/ PCP, but  not Capitella
capitata.  Molluscs markedly decreased at 7 yg/; annelids  and arthro-
pods at 76 yg/.  Few macrofauna occurred at 622 yg/.   Total  number of
animals and species were significantly less ( = 0.01) in aquaria ex-
posed to 76 yg/& than in those unexposed or exposed to 7 yg/.   There-
 fore, pentachlorophenol altered the structure of experimental  macro-
 fauna communities that developed during exposure by changing the rela-
 tive abundance of animals by species and phylum.

     Meiofauna community structures were studied in the  same tanks  as
 the macrofauna.  Exhibit 5 summarizes data  on the basis  of  rarefaction.
 Although there are structural changes in the community,  this is  not
 demonstrated by any of the species diversity indices shown:  "H" desig-
 nates Shannon-Weaver Composite index,-  "J,"  equibility; and  "SR,"
 species richness.

     Exhibit 6 shows the shift in nematode  feeding types as a  function
 of pentachlorophenol concentration.  An analysis of the  species  revealed
 that the meiofauna population shifted to predominantly detritis  feeders
 in 622  yg/A of pentachlorophenol.  The symbols used are:  1A,  selective
 deposit feeders; IB, nonselective deposit feeders; 2A, epigrowth feed-
 ers; and,  2B, omnivores.  Therefore, the functional types of meiofauna
 shift as a fraction of PCP concentration.

     This  experiment indicates that the test system described  with  meta
 oans may have distinct advantages; they have a relatively short  life
 cycle;  adults normally will reproduce continually; they  invade the  sys-
 tem rapidly.  Therefore, structure and functions of communities  can be
 determined.

     Exhibit 7 summarizes the aerobic microbial colony forming units
 observed in the presence and absence of pentachlorphenol.   The number
 of cultivatable microorganisms was significantly lowered in aquaria
 with high pentachlorophenol concentrations.
                                165

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  90
  75
  60
u_
o
CE-
Ul
CD
5
13
  45
  15
       ANNELIDS
                       MOLLUSKS
                                       ARTHROPODS
                                                       OTHER
                                                       PHYLA
         0  7 76 622    07 76 622    0  7  76 622    07  76 622
                      CONCENTRATION  (yg/liter)
EXHIBIT 4:  SUMMARY OF 6-WEEK MACROFAUNA COMMUNITY DIVERSITY
  STUDY WITH PENTACHLOROPHENOL
                               RAREFACTION
          16-
                                            H   J  SR
                                CONTROL  1.853  .72  1.97
                               -- 7ug/l     1.992  B3  1.68
                               - 76uq/l    1.864  .71  1.97
                               	 622ug/l   1.555  .65  2.14
                50     100    150    ?OO   250

                    NUMBER OF INDIVIDUALS
                                                300
350   400
     EXHIBIT  5:   SPECIES  DIVERSITY INDEX OF  NEMATODES
       EXPOSED TO VARIOUS CONCENTRATIONS OF  PCP
                             166

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  lOOr
   80
Q.
Q
LJ
LJ
O
o
   60h
   40
<  20
                           _L_

       IA  IB  2A  28   IA  IB 2A 2B   IA  IB  2A 2B   IA  IB  2A 2B

        CONTROL        7ug/l          76ug/.l        622 ug/l
                EXHIBIT 6:  EFFECT OF PCP ON

                  MEIOFAUNA COMMUNITIES
               Microbial  Colony Forming Units
    Control


    ~x~  15.93  x 10E


    a  10.75
500 ppb Pentachlorophenol


6.18 x 105


4.81
               t     = 2.85
               exp

               t     =2.26 (95% confidence)
               exp
       EXHIBIT 7:  EFFECTS OF PENTACHLOROPHENOL

              ON MICROBIAL POPULATIONS
                          167

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     Exhibit 8 is a picture of control grass shrimp (Palaemonetes pugio)
that have not been exposed to barite.  Exhibit 9 shows barite-exposed
shrimp with the intestinal tract (white)  filled with barite:  the shrimp
ingested the material.
SUMMARY AND DISCUSSION

     Data on the effects of chemical use in offshore well-drilling
operations are very limited.  Policy decisions are currently based on
static, 96-hr LC5Q determinations, observations of divers, and theoret-
ical models of pollutant dispersion.  A limited research program on
drilling-fluid constituents has been initiated at the Gulf Breeze Lab-
oratory to provide a better data base as one component in the evalua-
tion of the relative hazard of using alternative drilling mud constitu-
ents and to develop better laboratory methods for xenobiotic evaluation.
It is obvious that these methods should include:  effects on the struc-
ture and function of communities, indirect effects of pollutants, bio-
accumulation potential, toxicity of mixtures to organisms indigenous to
lease areas, attraction of marine species to chemicals used in well-
drilling operations, and mechanism of action of toxicants.  In this way,
choices among the multitude of alternative chemicals available for well-
drilling operations can be based on better toxicological information.
Hard data on effects might also eliminate unjustified concern about
effects of certain chemicals, which, in fact, may be safely used in
well-drilling operations.

     Exhibit 10 itemizes the study elements and the principal investi-
gators for the first half year of research on chemical use in offshore
well-drilling activities.
                                168

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                                                                                     01
                                                                                     VD
                                                                                     H
EXHIBIT 8:  CONTROL  GRASS  SHRIMP (PALAEMONETES PUGIO)

-------
                                                                                          o
EXHIBIT 9:  GRASS SHRIMP  (PALAEMONETES PUGIO) EXPOSED TO BARIUM  SULFATE

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  EXHIBIT 10:  LIST OF STUDY ELEMENTS, PRINCIPAL  INVESTIGATORS, AND
   GRANTEES DURING THE FIRST HALF YEAR OF  RESEARCH ON CHEMICAL USE
               IN OFFSHORE WELL-DRILLING ACTIVITIES
Study Element
Investigator
Affiliation
Planning Workshop
Chemical Analysis
Chemical Analysis
Statistical Analysis

Atomic Adsorption
Spectrescopy
Static Acute Toxicity
Static Acute Toxicity
Static Acute Toxicity
Flowthrough Toxicity
Flowthrough Toxicity
Macrofauna
Macrofauna
          
Macrofauna
Meiofauna

Microbiology

Biochemistry

Physiology

Physiology

Pathobiology
Photography and
Illustrations
M. Summers
J. Moore
A. Wilson
J. Ogelsby

A. Brennan

P. Borthwick
S. Schimmel
J. Patrick
S. Schimmel
J. Patrick
M.E. Tagatz
J. Ivey
M. Tobia
F. Cantalmo

E. Suarez

F. Fox

R. Rao

A. Cantalmo
S. Foss
University of Texas
EPA, Gulf Breeze
EPA, Gulf Breeze
University of West
Florida
University of West
Florida
EPA, Gulf Breeze
EPA, Gulf Breeze
EPA, Gulf Breeze
EPA, Gulf Breeze
EPA, Gulf Breeze
EPA, Gulf Breeze
EPA, Gulf Breeze
EPA, Gulf Breeze
University of West
Florida
University of West
Florida
University of West
Florida
University of West
Florida
University of West
Florida
EPA, Gulf Breeze
                                171

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 REFERENCED

 Sheen Technical Subcommittee.   1976.   Environmental Aspects of Drilling
   Muds and Cuttings from Oil and Gas  Extraction Operations in Offshore
   and Coastal Waters,  !_: 1-50.

 Fisher, P.  1975.   Conf. Proc.  of the Office of Toxic Substances, EPA
   Environmental Aspects of Chemical Use in Well-Drilling Operations,
   May 1975, Houston,  Texas,  EPA 560/1-75-004,  I_: 1-604.

 Wright, T.R., Jr.   1975-  "World Oil's 1975-1976 Drilling Fluids File,"
   World Oil, _!: 38-71.

 Working Group on Health and Environmental Effects of Energy Use.  1974.
   Report of the Interagency Working Group on Health and  Environmental
   Effects of Energy Use, prepared for the Office of Land Management and
   Budget, !_: 1-643.

 Hansen, D.J.  1974.  "Aroclor 1254:  Effect on Composition of Developing
   Estuarine Animal Communities  in the Laboratory," Contributions in
   Marine Science,  8_:  19-33.

 U.S.  Department of Interior.  Draft Environmental Statement Proposed
   1976.  Outer Continental Shelf Oil  and Gas Lease Sale  Gulf of Mexico.
   PCS Sale No. 44.  Prepared by the Bureau of Land Management.
 DISCUSSION

      Dr. Baumgartner:  You  stated that drilling muds are pumped over-
 board and released  at the base of the platform.  Is that the general
 practice or  is  some of this recycled?

      Dr. Richards:  You have to make a distinction here between the
 drilling operation  and production.  During the drilling phase, some of
 these materials are recycled.  Some of the Gulf of Mexico environmental
 impact statements summarize the tonnage data on the amounts that actu-
 ally  go overboard.

      Dr. Baumgartner:  Is it fair to ask how many tons per day are
 discharged?

      From the floor;  It depends on the tonnage of the well.

      Dr. Richards:  A table in Lease 47 Environmental Impact Statement
 gives  that.

     From the floor:  Do you know how much biocide is used to cut the
rust impact on the structures?
                                172

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     Dr.  Richards:  At the Houston  conference, held  by EPA about  a  year
ago, the types and quantities of biocides used were  summarized.   Bio-
cides are occasionally used to inhibit microbiotic growth,  which  causes
corrosion.  This poses a particular problem  for  the  EPA because of  dis-
charge permits from the platforms.  I do not have the data on  the quan-
tities used to minimize the bacterial growth in  the  gas production, but
there are figures on the other uses.  One other  point I might  also  make
is that barium sulphate is used as  a weighting agent, and  the  barium
sulphate helps the growth of several bacteria, which in turn,  produce
corrosive end-products.  To keep down the bacteria,  the biocides  must
be added.

     Dr. MeGammon:  Are you planning to look at  any  other  metals, such
as chromium?

     Dr. Richards:  As a matter of  fact, I have  only discussed a  small
amount of the things that we have already screened.  I wanted  to  give
one  example of something that was supposed to be totally negative and
one  that I thought was positive.  But,  in fact,  we have screened  chro-
mium sulphanate,  and we will be going into the community studies  with
that compound shortly.  We  are interested in that particular compound
because it has a  heavy usage pattern in the  Gulf of  Mexico.  In addi-
tion,  I have heard from one of the  drilling  mud  salesmen that  The
Netherlands has put some restrictions on its usage in the  North Sea.

     Dr. McCammon:  Are you going to try and define  the chemical  form
of  the metals, because I hear  that  it is hexavalent  chromium that is
coming out?

     Dr.  Richards:  As you  noticed  in that pyramid,  we had mechanisms
at  the top.  We are attempting to define the mode of action.   If  we can
understand the mechanism, then we can extrapolate to different organ-
 isms and  to different  situations.

     From the floor:   I do  not believe  that  EPA  exerts any control  over
drilling  rights.I think we only have  jurisdiction  over oil.  The
General Council has not ruled  on whether or  not  we have anything  to say
per se in terms of MBES drilling permits.
                                 173

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    THE CHEMISTRY AND ECOLOGICAL EFFECTS OF CHLORINATION OF SEAWATER:
                   A SUMMARY OF EPA RESEARCH PROJECTS

                  W.P. Davis,* D.P. Middaugh,* J.H. Carpenter,1"
                      G.R. Helz,** and M.H. Roberts, Jr.1"1"
     The use of chlorination to disinfect for bacterial control or con-
trol biological fouling is a widespread process unique to the United
States.  Many countries, including Sweden, Norway, West Germany, Great
Britain, and France, have been very conservative in the use of chlorine
for these applications.  Recent surveys have linked fish mortalities
and possibly shellfish bed recoveries to excessive chlorination or re-
duction of chlorination, respectively  (Virginia State Water Control
Board, 1974; Virginia Institute of Marine Science, 1975).  Studies by
Jolley  (1973) , Helz  (in manuscript), and Carpenter  (1976, 1977) reveal
that both halogenated and nonhalogenated compounds are produced from
chlorination of waters rich in organic compounds from natural produc-
tivity.

     In this paper,  we summarize  the results of ongoing research proj-
ects from 1975.  In  it, we relate preliminary findings to needs for fur-
ther investigation of the effects of chlorination on marine ecosystems.

     The annual production of chlorine in the United States was esti-
mated  as 95 x  Id11 kilos  (10.5 million tons) in 1975  (White,  1976).
The chemical industry uses 80 percent; the pulp and paper industry uti-
lizes  16 percent; 3  to 4 percent  is used for disinfection, including
municipal treatment, household uses, antifoulant in cooling waters,
swimming, and potable water treatment.  A recent estimate states that
200,000 tons of chlorine are used in disinfection.  Among the  50 states
 *  Environmental Protection Agency,  Bears  Bluff  Field  Station, P.O.
 Box  368, Johns  Island,  South  Carolina  29455.

 t  Division of  Chemical Oceanography,  School  of  Marine and Atmospheric
 Sciences, University of Miami,  4600  Rickenbacker Causeway, Miami,
 Florida  33149.

 ** University of Maryland, Box  106,  Chemistry Building,  College Park,
 Maryland 20742.

 tt Virginia Institute of Marine Science, Gloucester  Point, Virginia
 23062.
                                175

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surveyed none plan any reduction in these chlorination practices.  No
statistical data seem available to estimate the proportions of usage
directly or indirectly affecting marine waters.  It is reasonable to
state that the high "chlorine demand" of coastal waters results in high
utilization wherever these waters are used as cooling for power plants.
Municipal treatment plants also contribute a substantial, if not greater,
level of chlorination to estuarine and coastal waters.

     Since 1975, the Gulf Breeze Environmental Research Laboratory,
Bears Bluff Field Station, near Charleston, South Carolina, has been
coordinating a study of the marine aspects of chlorination.  This effort,
with extramural grants supported through Environmental Protection
Agency's  (EPA) Office of Energy Minerals and Industry (OEMI),  is de-
signed to establish a scientific basis for decisions guiding future
energy production and usage patterns, and environmental protection mea-
sures.
CHEMISTRY OF CHLORINATION OF
MARINE WATERS	

     Marine waters are characterized and defined by the levels of dis-
solved halides, principally chloride and bromide.  Unlike the situation
in fresh water, strong oxidants added to marine waters react nearly in-
stantaneously with natural bromide and produce oxidative states of bro-
mine, including hypobromous acid, hypobromite ion, and bromate
(Carpenter and Macalady, 1976; Carpenter ejt a^. , 1977).  These findings
are significant because the most popular method currently used to esti-
mate "residual chlorine" does not provide accurate determinations of
hypobromite; therefore, the input to the environment is seriously under-
estimated (Carpenter et aj^. , in manuscript) .  In addition, this ampero-
metric titration method does not sense bromate at all; yet this form of
bromine may persist in marine environments for long periods of time with
unknown ecological effects (Carpenter et al., 1977).

     Additionally, chlorination of sea water produces reaction products
from halogenation of both natural and man^-produced organic compounds.
Natural sea water chlorinated in the laboratory produces chloro-bromo
analogs not found in unchlorinated sea water (Carpenter, 1977; Carpenter
and Macalady, 1976) .

     Analyses and examination of these reaction products required mod-
ification and development of demanding techniques.  Four major steps
are involved:

1.   Isolation and concentration of the organic fraction from the bulk
     aqueous phase

2.   Separation of the organic fraction into individual compounds

3.   Detection and characterization of each compound
                               176

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4.   Final identification of each compound.

     Separation and characterization of compounds trapped from the iso-
lation and concentration step are performed by a combination of gas
chromatography and mass spectrometry methods.  Since as many as 150
compounds may be separated in a single run, a minicomputer system is
used to accumulate the copious data produced.  Organic compounds are
identified by reference to laboratory standards and to computer librar-
ies, where the mass spectra may be compared to the over 41,100 compounds
on file.

     These investigations, conducted at the School of Marine and Atmo-
spheric Sciences, University of Miami under Dr. James H. Carpenter, have
isolated a large number of compounds and tentatively identified 20 of
these.  Chlorination produced bromoform, bromoxylene, bromomesitylene,
dibromochlomethane, and chloroform.  Many other compounds are obviously
present and are being investigated using high-resolution capillary col-
umns.  Chlorination of sea water exposed to sunlight increases forma-
tion of halogenated organics.  In the case of bromoform, sunlight in-
creases production approximately tenfold, yielding concentrations of
several hundred ppb  (Carpenter, personal communication).

     Analyses and inventory of these by-products are continuing.  Forma-
tion of halogenated compounds with low solubility is being examined with
an eye to the entry of these forms into marine food webs, as previously
known for many pesticides.

     Investigation into the detection of compounds from chlorination in
the cooling water effluent waters from a power plant is one of the prin-
cipal emphases of a research grant to the University of Maryland, under
the direction of Dr. George R. Helz.  These investigations have detected
bromoform in the chlorinated cooling effluent waters.  Furthermore, it
has been demonstrated that chlorination of waters with natural "humic
acids" can produce nitriles such as hydrogen cyanide  (Hsu and Helz, in
manuscript).  Further work must be performed to better define the pro-
duction rates of both these compounds.

     The methodology for testing potential biological uptake and accumu-
lation is being modified from work on pesticides.  Still, bromoform and
cyanide pose special problems since little experience exists for tissue
analysis in aquatic organisms for these compounds.
 ECOLOGICAL  IMPACT  OF  CHLORINATION
 ON MARINE ORGANISMS	

      Much biological  work  is  yet to  come  as  the  chemical characteriza-
 tion and identification  tasks continue.   Initial phases of biological
 investigations  involved  compilation  and review of available data on
 toxicity of chlorination (Davis and  Middaugh, 1976; Middaugh and Davis,
 1976).   Published  results  were typically  expressed as the toxicity of


                                177

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so-called "total residual chlorine," which as we previously mentioned
(Carpenter et al.,  in manuscript),  is technically and scientifically
incorrect for marine waters.  It is immediately apparent that far less
information exists  for biological effects of marine water chlorination
compared to fresh water situations  (Brungs, 1973; Davis and Middaugh,
1976; Jolley, 1976).

     To achieve close communication of ongoing research projects, a
workshop was held at Chesapeake Biological Laboratory.  Here, investiga-
tors agreed that "chlorination-induced oxidants" should be the proper
terminology used for active oxidants affecting the readings achieved on
amperometric titrators.  Additionally, 15 papers were presented that
summarized the state of the art for inorganic and organic chemistry,
biological toxicology, and physiology of marine waters  (Chesapeake
Science, 10).

     Toxicological studies have been carried out both extramurally at
the Virginia Institute of Marine Science on mollu'scs and decapod crus-
taceans and with fishes at Bears Bluff Field Station.  Investigations
on decapod crustaceans have established toxicity data for larval crabs
and for the bivalve mollusc Mulinia lateralis.  Dr. Morris Roberts has
also directed investigations on the seriological effects on the blue
crab (Callinectes sapidus) and design of a behavioral test for migrat-
ing larval decapods.  One area originally proposed, the impact upon
marine plants (Zostera), has been indefinitely postponed until the
chemical definition of chlorination by-products is more complete.

     Investigations carried out at Bears Bluff Field Station include:

1.   Variations of  "chlorine demand" in natural waters over the tidal
     cycle  (Crane e_t a_l_., manuscript in preparation).  In this analysis,
     chlorine demand variations are clearly related to tidal stage, with
     a dozen parameters being tested statistically to clarify the role
     of principal factors and their interactions.

2.   "Free Adenosine Tri-phosphate  (ATP)" from the sea water system,
     marine ecosystem testing units, and marsh ponds have been tested
     (Erickson ejt al^., in manuscript).  The marine ecosystem testing
     units  (METU) provided nonchlorinated controls and three levels of
     low chlorination for comparison.  The ATP levels were clearly de-
     pressed according to the chlorination level, with the implication
     that productivity may be affected (Gentile et al., 1976).

3.   Several studies have concerned the effects of chlorination upon
     early life history stages of fishes  (Middaugh e_t al_. , 1977;
     Middaugh et al., in manuscript).

     In studies of the striped bass  (Morone saxitilis), percent of egg
     hatchability,  incipient LC50 bioassays, histopathological effects,
     and behavioral avoidance were used to test response to chlorina-
     tion (Middaugh ejt aJU , 1977).   Developing embryos were exposed


                                178

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     8-9 hr after fertilization until normal time of hatching to a se-
     ries of low levels of continuous chlorination rates, as low as
     0.01 mg/Jl.  Histological examination of surviving 30-day-old juve-
     niles from incipient LC50 bioassays demonstrated gill and pseudo-
     branch damage in individuals at 0.21 to 2.36 mg/ in chlorination-
     induced oxidant levels.

     Avoidance tests using 24-day-old juvenile striped bass exhibited
     experimentally reproducible responses at chlorination-induced oxi-
     dant levels between 0.29-0.32 mg/&, and 0.79-0.82 mg/; however,
     they failed to respond to levels between 0.16-0.18 mg/.

4.   The marine fish 'spot  (Leiostomus xanthurus) was tested in the juve-
     nile stages  (Middaugh et al., in manuscript).  The spot showed a
     temperature-dependent avoidance to chlorination.  At test tempera-
     tures of 15 and 20c, fishes exhibited avoidance at lower chlori-
     nation levels than during tests at 10C.  Exposure of spot previ-
     ously acclimated at 13C to simultaneous stresses of chlorination
     and temperature increases (A+ 5, 10, 13C) exhibited increased
     sensitivity at the highest temperature.

5.   Factorial analyses were performed using different developmental
     stages of the mummichog  (Fundulus heteroclitus) at four chlorina-
     tion levels, and three temperature levels during four time inter-
     vals  (Middaugh et al., in manuscript).  These efforts showed that
     1 to 7-day-old larvae were much more sensitive than embryos.  The
     chorionic membrane of the egg of this species seems to afford
     protection to the embryonic stages.

6.   The impacts of low-level chlorination  (<1.0 ppm) upon the condition
     of the oyster  (Crassostrea virginica) are currently being tested.
     Survival, feeding response, fecal pellet production, tissue respi-
     ration, reproductive success, and larval response are being investi-
     gated.  It is interesting to note that larval oysters settle only
     in non-chlorination controls in METU tests  (Scott and Middaugh, in
     manuscript; Scott and Laurence, in manuscript).

7.   Larval organisms entrained in the sea water system are settled in
     38 Si METUs and are being systematically sampled  (Davis et al.,
     1977).  Three levels of continual chlorination are tested  (nominal
     levels of 0.50, 0.25, and 0.125 ppm).  These microcosms are main-
     tained outdoors and exposed to natural conditions of lighting and
     precipitation.  Distinct effects have been  seen in algal popula-
     tions, sediment characteristics, and among  selected invertebrates
      (e.g., oysters and barnacles) to chlorination levels.
                                179

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COORDINATION OF MARINE CHLORINATION
RESEARCH RESULTS	.

     It is obvious from the ,broad spread of the aspects of.chlorination
in marine waters that the needs of EPA and other decision makers cannot
be met solely by a modest research program and a collection of well-
designed scientific projects.  The key is comparison of results and com-
munication among a wide variety of researchers.  We have been fortunate
that two symposia are timed to accommodate the rapid progress in the
marine section of chlorination research.  The first symposium, Environ-
mental Impact of Water Chlorination held in 1975 has been published
 (R. Jolley, ed., 1976), and the second symposium is scheduled for early
November 1977.  These symposia will serve to bring the total chlorina-
tion research and application literature up to date.  The forthcoming
symposium serves as a milestone for publication of the scientific out-
comes of each of the EPA/OEMI extramural grants and Gulf Breeze Environ-
mental Research Laboratory/Bears Bluff Field Station in-house projects
as well.  The EPA/OEMI workshop held at Chesapeake Biological Laboratory
provided an intermediate step to maintain communication among the
 "marine" section of chlorination research.  It is clear that research
activities at this point have proceeded from the initial emphasis upon
toxicological effects of sea water chlorination per se to the defini-
tion of the roles and effects of various by-products resulting from
chlorination practices.

     Other oxidizing agents have been proposed as replacements for chlo
rination, including bromochloride and ozone.  However, in marine waters,
we now know that release of bromine is the key feature, and it is clear
bromine is released by both of these  (alternative) chemical oxidizing
agents.  Therefore, chlorination practices must be carefully examined
on the basis of real need.

     Practices in England are distinctly different than in the United
States  (Coughlin and Whitehouse, 1977).  Adjustment of chlorination to
achieve avoidance by fouling organisms would drastically reduce current
chlorination levels in specific cases.  In certain tropical areas,
chlorination has been dropped altogether, since in these situations the
thermal increase alone prevents biological fouling.  The role of de-
chlorination must be carefully examined as an option, where antifouling
is demonstrated to be uncontrollable by other means.  Elimination of
power plant chlorination may be achieved by simply requiring the need
for chlorination to be clearly demonstrated as the only option available.

     Chlorination for disinfection of effluents from municipal treatment
plants entering marine environments seems to be a need based on myth or
fear of pathogens, since it is principally in the continental United
States that this destructive practice is maintained.  Furthermore, it is
far from certain that chlorination, in fact, controls viral pathogens
nearly as well as the indicator bacterium E_. cpli used to assess munic-
ipal effluents.   For thousands of years, man in various places and among
many cultures has realized that fresh water occasionally transmits
                               180

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disease by direct contact  (other  than  drinking),  but rarely  are marine
waters implicated in pathogenic transmission.   The  idea that we must
disinfect well-processed municipal  effluents  entering marine waters is
basically absurd.  Standards must address  the  problem rather than con-
tinue to perpetuate a  convenient  myth.
REFERENCES

Block, R.M., G.R.  Helz,  and W.P.  Davis.   1977.   "The  Fate  and  Effects
  of Chlorine  in Coastal Waters," Ches.  Sci.  18_ (1):   97-101.

Brungs, W.A.   1973.   "Effects  of  Residual Chlorine on Aquatic  Life,"
  J. Water  Poll. Control Fed.  45  (10):   2180-2192.

Carpenter,  J.H.  1977.   "Problems in Measuring  Residuals in Chlorinated
  Seawater," Ches.  Sci.  18_ (1):   112.

Carpenter,  J.H. and D.L. Macalady.   1976.  "Chemistry of Halogens  in
  Seawater."   In_ R.  Jolley, ed.,  Proc.  Conf.  Environmental Impact  of
  Water Chlorination,  ORNL Conf.  751096:  177-193.

 Carpenter,  J.H.,  D.L.  Macalady,  and C.A. Moore.  1977.  "Sunlight
   Induced Bromate  Formation in Chlorinated Seawater," to be published
   in Science.

 Carpenter,  J.H., C.A.  Moore, and  D.L. Macalady.  "Errors in Determina-
   tion of Residual Oxidants in Chlorinated Seawater."  Manuscript.

 Crane, A.M., L.H.  Bahner, and R.G.  Domey.  "Influences of  Selected Tidal
   Activity Dependent Parameters Upon Chlorine Demand  in an Estuarine
   Environment."  Manuscript in preparation.

 Coughlin, J. and J.  Whitehouse.   1977.   "Aspects of Chlorine Utilization
   in the  United Kingdom," Ches.  Sci. 1(3  (1):   102-111.

 Davis, W.P,, B.S.  Hester, R.L. Yoakum,  and R.G. Domey. 1977.   "Marine
   Ecosystem Testing Units:  Design for  Assessment of  Benthic Organism
   Responses to Low-Level Pollutants," Int. Symp. Biologische Anstalt
   Helgoland, Helgoland wiss, Meeresunters.

 Davis, W.P. and D.P. Middaugh.  1976.   "A Review of the Impact of  Chlo-
   rination Processes Upon Marine  Ecosystems."  In R.  Jolley, ed.,  Proc.
   Conf. on Environmental Impact of Water Chlorination, ORNL Conf.
   751096:  299-325.

 Dove,  R.A.   1970.   "Reaction of Small Dosages of Chlorine  in Seawater."
   Electric Generating Board, Southeastern Region, Research Report  42/70
   File No.  0.3070/ID Job 10665,  189 pp.
                                181

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Erickson, S.J. and H.R. Foulk.  "The Effects of Continuous Chlorination
  on Entrained Estuarine Plankton as Measured by Adenosine Triphosphate."
  Manuscript.

Gentile, J.H., J. Cardin, M. Johnson, and S. Sosnowski.  1976.  "Power
  Plants, Chlorine and Estuaries," Env. Prot. Agy. Ecol. Res. Ser. EPA-
  600/3-76-055, 28 pp.

Helz, G.R., R.Y. Hsu, and R.M. Block.  "Bromoform Production by Oxida-
  tive Biocide in Marine Waters."  Manuscript.

Hsu, R.Y. and G.R. Helz.  "Organic Cyanides in Estuarine Power Plant
  Discharges:  A Non-Halogenated Product of Chlorination."  Manuscript.

Jolley, R.L.  1973.  "Chlorination Effects on Organic Constituents in
  Effluents from Domestic Sanitary Sewage Treatment Plants," PhD Disser-
  tation, University of Tennessee, Knoxsville.  339 pp.

Jolley, R.L. Ed.  1976.  The Environmental Impact of Water Chlorination;
  Proc. Conf. on the Environmental Impact of Water Chlorination,  ORNL
  Conf. 751096, UC-11, 41, 48.  443 pp.  (Available from National Tech-
  nical Information Service, U.S.  Department of Commerce, Springfield,
  Va. 22161.)

Jolley, R.L.  1977.  "Identification of Organic Halogen Products,"
  Ches. Sci. 1B_  (1):  122-125.

Middaugh, D.P. and A.C. Badger.  "Laboratory Spawning and Development of
  the Spot, Leiostomus zanthurus Lacepede," Aquaculture.  In Press.

Middaugh, D.P., J.A. Couch, and A.M. Crane.  1977.  Responses of Early
  Life History Stages of the Striped Bass, Morone Saxatilis to Chlori-
  nation," Ches. Sci. 1  (1):  141-153.

Middaugh, D.P., A.M. Crane, and J.A. Couch.  "Toxicity of Chlorine to
  Juvenile Spot, Leiostomus xanthurus,"  Submitted to Water Research.

Middaugh, D.P., and W.P. Davis.  1976.   "Impact of Chlorination Pro-
  cesses on Marine Ecosystems," Proc. of EPA/AIBS Symposium, EPA 600/3-
  76-079):  46-62.

Middaugh, D.P., J.M. Dean, R.G. Domey,  and G. Floyd.  "Effects of Ther-
  mal Stress and Total Residual Chlorination of Early Life Stages of
  the Mummichog, Fundulus heteroclitus."  Manuscript.

Scott, G.I. and D.P. Middaugh.  "Preliminary Study in the Toxicity of
  Effective Chlorination to the American Oyster, Crassostrea Virginica."
  Manuscript.
                               182

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Scott, G.I. and D.R. Lawrence.   "Oyster  Condition Index as  a  Monitor  of
  Biological Pollution  in  South  Carolina Coastal  Waters."   Manuscript.

Sprague, J.B.  1973.  "Measurement of Pollutant Toxicity to Fish.   Bio-
  assay Methods for Acute  Toxicity,"  Water Research 3:  793-798.

Virginia Institute of Marine Science.  1975.   Proceedings of  a Seminar
  Held by  the  Select Inter-Agency Task Force  on chlorine, 140 pp.

Virginia State Water Control Board.  1974. James River Fish  Kill.
  73-025.  Bureau of Surveillance and Field Studies,  Division of
  Ecological Studies, 61 p.

White, G.C.  1976.   "Current Chlorination and Dechlorination  Practices
  in  the Treatment of Potable Water,  Wastewater,  and  Cooling  Water,"
  In  R. Jolley, ed., Proc. Conf. on Environmental Impact of Water
  Chlorination, ORNL Conf. 751096:  7-24.
 DISCUSSION

      From the floor:   I have to take exception to that statement you
 just made.   Dr.  Cavelli has been conducting an epidemiclogical  study
 of  bathers  at Rockaway and Coney Island.   He has been correlating  the
 incidence of gastrointestinal upset with coliform levels  and  other indi-
 cators in the swimming water.

      From the floor;   Would funding of research to derive engineering
 approaches  to dechlorination be a wiser policy for the EPA than an
 ecotex program?

      Dr.  Davis;   Originally, we were studying this purely from  the
 point of  view of the  oxidizing effect on the organism. I think, at
 this point, that we have demonstrated that this issue is  secondary to
 assessing the pathways of the induced halogenated by-products.  I  am
 not sure  that dechlorination is the only alternative.

      Dr.  Carpenter;  There was some preliminary work at the University
 of  Miami  showing that ferrates are effective biocides and that  the end
 product is  just  ferric iron, a relatively innocuous material  to return
 to  the environment.

      Dr.  Davis:   I think the problem with the pathogens and incidence
 of  disease  in recreational waters is that you are trying  to correlate
 it  with coliform.  Until we correlate it with the actual  pathogen,
 which is  the basis of the problem, we are always going to be  whistling
 in  the dark.

      Dr.  Baumgartner;  I think that dechlorination will probably work
 for some  power plants.  For municipal sewage discharges,  it depends on
 how well  the sewage is treated, how the dechlorination step is  done,
                                 183

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and how soon it is done.  If chlorinated organics are formed, some of
the conventional dechlorination steps are just going to remove the free
chlorine.

     Dr. Davis:  You are just going to reduce free chlorine and not
affect the halogen.

     Dr. Baumgartner:  If it is a poorly treated sewage, the chlorinated
organics, in the long run, may be a much greater problem than the free
chlorine residuals in the marine environment.

     Dr. Hergott;  Dechlorination in power plants is not easy, because
some kind of contact time is required.  At the power plants in Califor-
nia, there is approximately 1/2 min of flow time from the condensers to
the outflow, which is not long enough.

     Dr. Davis:  At least, you achieve a lot of mixing during that time.
Are the rates for reaction of dechlorination available?

     Mr. Hergott:  It depends on the form of the residual.

     Dr. Davis:  It does require reexamining each situation and redesign.
On the other hand, we are using a design basically set up by the munic-
ipal treatment supply industries, which^predicate everything on a chlo-
rine residual.  There is no real attempt on their part to find out the
necessary level of chlorination; or what they are preventing; and at
what time of year; and how much is needed to achieve it.

     Dr. BaUmgartner;  Maybe there is enough evidence to say that power
plants should not use chlorine or should use it in such a way that the
residual is 0.001.

     Dr. Riley;  Are you running any studies when you chlorinate at
higher temperatures?  For example, the cooling tower temperatures?

     Dr. Davis:  We are not trying to assimilate cooling tower pro-
cesses.  The other part of your question is answered by the fact that,
in the outdoor exposure, we are monitoring the seasonal temperature and
the temperature of the sea water continually-  In the laboratory, where
we are studying exposures of the individual species, we are working at
hi ghe r tempe rature s.

     Dr. Riley;  What are your future plans for looking at some of
these chlorinated organic compounds with regard to toxicity to marine
organisms?

     Dr. Davis;  There seems to be a distinct uptake by fishes, at
least, of bromoform.  We rather expected bromoform to be volatile and
disappear like your light oil fraction.  We can readily identify the
other compounds.
                                184

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     grom the floor:   I have  one  question.   Have  you looked at any
pelagic fish eggs,  relative to  the  benthic  ones?

     Dr. Davis;  To compare them?  No.   If  you  are  thinking about
higher sensitivity  of  a pelagic fish egg, I would agree.

     From the floor:   We  found  that there is a  difference,  for example,
between the menidia and spot, with  regard to count.   One  will  affect
the development, and the  other  one  will  not.

     Dr. Davis:  In this  case,  the  menidia  is probably resistant.
EPA's alternative prime fish  species lays its eggs  intertidally.

     From the floor:   Have you  heard anything about this  chlorine  flux
monitor that may be produced?

     Dr. Davis;  In essence,  I  do not think it  is any different than an
aperometic titrator.

     Dr. Carpenter: The  flux monitor has the same  basic  chemistry as
the titrator, only  the time of  reaction  is  probably 10 or 15 sec.  be-
tween the point of  injection  of the potassium iodide and  the electrodes.
However, you are just  not going to  get quantitative results.   You  will
severely underestimate the actual residual  oxide  with the flux monitor.

     Dr. Riley;  What  is  the  GC mass spectroscopy technique that has
been used to identify  these chlorinated  organics?

     Dr. Carpenter: Basically, it  is the Hewlitt-Packard system.

     Dr. 'Anderson:   You mentioned something about the eggs  being per-
haps the most resistant or most sensitive.

     Dr. Davis:  More  resistant than newly  hatched.   We are examining
the stage life history against  different temperatures and different
doses.  The particular case with  coreanic membrane  just indicates  rela-
tive sensitivity at life  history  stage.

     Dr. Anderson:   That  is certainly consistent  with the hydrocarbon
findings.  However, we found  that it was not necessarily  the coreanic
membrane, because  I had one person  who actually dissected the  membrane
off of a number of  embryos and  the  tolerance was  the same.

     Dr. Davis:  What  was the organism?

     Dr. Anderson:   It was a  fungilus, either a heteroquitous  or a
grandis.

     Dr. Davis:  We are trying  this with a  self-fertilizing hermaphro-
dite, to see if we  can get successful embryological development without
                                185

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the coreanic, and compare that.  I am interested that somebody has, at
least, done it.

     Dr.  Carpenter:   I just wanted to make one comment concerning new
regulations.  If regulations are arbitrarily restrictive, you are driv-
ing people to some other technology, probably mechanical, a la Amortat,
which could increase the copper emission rate.  So, we need to examine
whether we would rather have copper or chlorine before deciding what
should be done.  I think it is a little bit early to determine the
magnitude of chlorine environmental impact.
                               186

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     POWER PLANT COOLING WATER CHLORINATION IN NORTHERN CALIFORNIA

                    S. Hergott, D. Jenkins, and J.F. Thomas*
     Power plants require large volumes of cooling water to carry away
waste heat.  If fouling is not controlled, bacterial and algal slimes
attach themselves to the walls of the piping and decrease the heat
transfer across the condensers.  The most economic method currently
in use to control fouling is the addition of chlorine to the cooling
water.

     Chlorine presents problems when its toxic effects carry over into
the receiving waters.  Brungs  (1973) conducted a comprehensive review of
the literature dealing with toxicity of both continuous and intermittent
chlorine treatment to fresh water fish.  Based on these studies, he rec-
ommended intermittent chlorine limits in the discharge of power plants
that vary from 0.2 mg/ to 0.04 mg/& for up to 2 hr per day, depending
on the degree of protection desired.  Basch and Truchan  (1976) studied
the effect of intermittent chlorination on brown trout  (Salmo trutta)
at five Michigan power plants.  They found the 48 hr total residual
chlorine intermittent concentrations lethal to 50% (ILC-50) of the
brown trout to range from 0.14 to 0.17 mg/SL and 0.18 to 0.19 mg/ for
fish exposed to two and four 30 min chlorination cycles, respectively.
Mattice and Zittel  (1976) conducted a comprehensive review of the liter-
ature dealing with the toxicity of chlorine to both fresh water and
marine organisms.  Based on this research, they determined both acute
and chronic toxicity thresholds and concluded that marine organisms
appear more susceptible to acute doses of chlorine, whereas fresh water
organisms appear more susceptible to chronic concentrations.

     The increasing awareness of the toxicity of chlorine residuals has
prompted regulatory control of chlorine discharges in power plant co61-
ing waters.  The Environmental Protection Agency  (EPA) has established
free chlorine discharge guidelines of 0.5 mg/H maximum and 0.2 mg/
average concentrations for up to 2 hours per day from any one unit
(1974).  The California Central Coast Regional Water Quality Control
*    Sanitary Engineering Research Laboratory, College of Engineering
and School of Public Health, University of California, Berkeley,
California  94720.
                                 187

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Board has set daily maximum  limits  of  0.02 mg C12/& undissociated free
available chlorine and  0.1 mg  Cl2/ total  available chlorine in power
plant discharges  (1976).  The  San Francisco Bay Regional Water Quality
Control Board has set even stricter limitations of an instantaneous
maximum chlorine residual of 0.0 mg/&  (1976).

     This study was conducted  to determine the nature,  levels, and per-
sistence of chlorinated compounds in the discharges of  five power plants
in northern California.  In-plant studies  were conducted to understand
the demands for chlorine in  the cooling water system and to determine
the levels reaching the receiving waters.   Decay studies conducted at
the outfalls, together  with  measurements of oxidant residuals in the
receiving water, provide a base for estimating the persistence and
zone of influence of the chlorinated compounds.
MATERIAL AND METHODS

     We describe below the power plants and  their  location,  its chlorina-
tion practices, field study procedures, and  analytical methods and
equipment.

     Power Plant Location and Description

     Exhibit 1 is a map showing the  location of  the  five power plants
studied.  Water quality varied from  fresh  at Contra  Costa,  to marine at
Moss
                                                           DSACRAMENTO
                                  PACIFIC

                                  OCEAN'
                                           MONTEREY\
                                             BAY  kwOSS LANDING P.P.
EXHIBIT 1:  POWER PLANT
  LOCATIONS
                                               MONTEREY
 V
t
                                188

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     A summary of  the  operating data for each plant is presented in
Exhibit 2.  All  the  plants have the capability of burning either fuel
oil or natural gas.  The plants rarely operated at maximum capacity.
Usually, the newer,  more efficient units in any plant operated contin-
uously, and the  older  units were used during peak demand periods and
during periodic  shutdowns of the other units.
             EXHIBIT 2:   POWER PLANT OPERATING DATA
Power Plant Gross Generating No. of Source of Cooling Water
Plant Capacity (MW) Units Intake Water Flow Rate*
(m3/sec) (gpm)
Contra Costa 1300
7 San Joaquin 43.2
River
684 000
     Plttsburg
                                     Suisun Bay
                                                  45.4
                                                           720 000
Hunters
Point
Potrero
Moss Landing
400
330
2110
3
3
7
San Francisco
Bay
San Francisco
Bay
Monterey Bay
18.2
16.5
63.1
288 000
262 000
1 000 000
      * All units in operation.

      "I* Unit 7 cooled by a spray canal.
      Plan diagrams of the once-through cooling water systems at each of
 the power plants are presented in Exhibits 3 to 7.  Cooling water is
 diverted from its source and pumped through the cooling system by cir-
 culating water pumps located at the intake structures.  The cooling
 water first passes through bar racks, which prevent large objects from
 entering the system.  Smaller objects are removed by traveling screens,
 which rotate periodically and are cleaned with high pressure water.

      Heat exchange between cooling water and superheated steam occurs in
 the condensers,  which consist of bundles of 22.2 mm (7/8 in.) or 25.4 mm
 (1 in.)  diameter aluminum-brass or copper-nickel alloy tubes.  There are
 generally two condensers per unit.  The heated flows of cooling water
 combine  immediately after the condensers and are discharged to the
 receiving water at the outfall structure.

      In  Exhibits 3 to 7, cooling water sampling locations are Indicated
 for each power plant.  Cooling water could generally be sampled at the
 condenser inlets (COND-IN),  condenser outlets  (COND-OUT), and at outfall
 manholes (OUT).   Convenient sample taps had been set up at the COND-IN,
 because  this was where oxidant residuals were routinely measured by
 plant operating personnel.  Although taps generally existed or were
 readily  installed,  sampling at the COND-OUT was more difficult, because
 samples  had to be pumped out against a vacuum of about 25.4 cm (10 in.)
                                 189

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        SANJOAQUIN RIVER
      TURBINE
      GENERATOR
      BUILDING
      UNITS 6 & 7
 SAMPLING LOCATIONS
PLAN
0   100  ZOO  300  4OO FT
EXHIBIT 3:   CONTRA  COSTA POWER PLANT
  Cooling Water Intake Structures
               Un'"5s6 Unns 1,2.3.1
EXHIBIT 4:   PITTSBURG POWER PLANT
                 190

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                   EXHIBIT 5:   HUNTERS  POINT  POWER PLANT
                                 	 B|  -i.  if-""*'   j	 rntyKe Lonuuits un
Cooling Wtr.
Intake Structure
Unils 1.2,3
                                                                f ^Sampling Locations
                             SO  100  ISO   ZOO FT.

-------
EXHIBIT 6:   POTRERO POWER PLANT
Cooling Water
Intake Structure
Units 1 & 2
I make Structure Discharge Structure | 1 ^1
Unit 3 UnitS J I NewFi|,Area J
Injection Points fob Uj_ /Chlorine , _T Chlorine Line i j \y
TJ^ Building 5~ 1 	 . 1 (


Intake Pipes V S__/v ^. Conduit
Unit 3 ^ -^ - \j5) 	 r-l Stac
Turbine 1
Generator \
Buildinq A
Units' I H
Chlorine fa\
Injection Jfo \ Pump
Point t^To 1 '~'ouse
V
O Sampling Locations |
Intake
I














\ t
Pipes "1 \
I Units 142 1 Discharge
l| (structure

Turbine
Generator
Building
Units 1&2
Units 1 2
' 1 ''Discharge
'': Conduits
4*
                                         300  tOO FT.

-------
of mercury.  Outfall manholes  generally  existed  as part  of  a  discharge
structure so that samples taken from  these manholes were representative
of the water being discharged  to  the  receiving water.  At Moss  Landing,
Units 6 and 7 discharged into  Monterey Bay,  242  m  (800 ft)  offshore,
at a depth of 6.1 m  (20 ft).   A sampling point approximately  midway
between the condensers and the outfall was furnished by  International
Shellfish Enterprises, Inc.

     Chlorination Practices

     Chlorine is injected intermittently into the cooling water flow to
control slime-causing organisms.   In  Pacific Gas and Electric Company
 (PG&E) plants, liquid chlorine is withdrawn  from 1-ton cylinders, evap-
orated, and injected into a water line to produce a concentrated chlor-
ine solution.  This solution flows to the point  of injection  in a well
behind the traveling screens and  in front of the circulating  water
pumps.  Oxidant residuals are  routinely  measured at the  COND-IN, and
the chlorine injection rate is adjusted  to give  the desired residual.
These adjustments are rarely needed on a daily basis; however,  the re-
 sidual monitoring is important to ensure proper  operation of  the chlori-
nators.   Chlorinators are somewhat unreliable when subjected  to the fre-
quent startups and shutdowns encountered in  intermittent chlorination.

     A summary of chlorination practices for the newest  units at each of
 the five  PG&E power plants  is  presented  in Exhibit 8.  The  newest units
 were chosen for  study because  they were  the  largest and  most  frequently
 operated.  A great deal of  variation  in  frequency, duration,  and levels
 of chlorination  existed.  Frequency varied from  I/week to 4/day and
 was changed during the year to meet changing demands.  Duration of
 chlorination ranged  from  15 to 40 min.   All  the  plants seek to  maintain
 a 0.5 mg/ residual  at the  COND-IN except for Moss Landing, where a
 1.0 mg/  residual for a shorter  duration was used.  Hunters Point was
 the only  plant to measure a free  residual and use  it as  their criterion
 for dosing.  Until recently,  the  orthotolidine method was used  at all
 the plants, because  it was  the most portable and the easiest  for the
 operators to learn and use.  All  the  plants  are  currently using or are
 in the process of changing  over  to the amperometric titration method.

     Field Study Procedures

     Field studies were conducted at  four of the power plants.   Since
 no convenient sampling point existed  near the outfall, no studies were
 conducted at the Pittsburg  plant.  Studies were  conducted during normal
 chlorination cycles and were designed to monitor existing chlorination
 practices; therefore, no changes  were made in chlorine injection rate,
 duration, or frequency of chlorination cycles to accommodate  this work.

     At the beginning of the study, oxidant  residuals were  determined
 solely by the amperometric method. During the course of the  investiga-
 tion, the DPD-FAS titrimetric  method  was added in an attempt  to distin-
 guish between bromine and chlorine residuals.
                                193

-------
                     EXHIBIT 7:  MOSS LANDING  POWER PLANT
O Sampling Locations

-------
               EXHIBIT 8:  CHLORINATION PRACTICES AT PACIFIC GAS AND ELECTRIC PLANTS
Power Plant
Contra Costa
Pittsburg

Hunters Point
Potrero
Moss Landing
Unit(s)
6,7
5,6

4
3
6,7
Chlorination Schedule
Frequency
1 /week
Varies with season
Min-1/week
Max- 1 /day
4/day
2/day
I/day
Duration
(min)
40

30
30
30
30
15
Criteria
at
COND-IN
(mg C12A)
TOR=0.5
TOR=0.5

TOR=0.5f
TOR=0.5f
TOR=1.0
Analytical Method
Nov. 1975
Amp. Tit.
OT*

Amp. Tit.
OT*
OT*
Current/Future
Amp. Tit.
Amp. Tit.

Amp. Tit.
Amp. Tit.
OT/Amp. Tit.
*  OT = Orthotolidine Method.

t
                                                                                                       LD
                                                                                                       CTl
   Lowered to 0.5 mg/  in January  1977.

-------
     Cooling water samples were collected for the determinations of the
water quality parameters listed in Exhibits 9-11.  At the COND-IN, trip-
licate IX, grab samples were taken before, during, and after chlorina-
tion, both from a condenser being chlorinated and from one that was not.
Samples were taken at the outfalls during chlorination.  To destroy oxi-
dant residuals, a small excess of sodium arsenite was added to all sam-
ples taken during chlorination.  No interference from this procedure
was found in any of the analytical methods, but there was some evidence
that the sodium arsenite did not always react instantaneously with the
oxidant residual.  This would probably result in lower concentrations
of NH3~N and organic nitrogen in those samples taken during chlorina-
tion.

     Decay studies were conducted on chlorinated cooling water collected
at the outfalls.  Two 8X, (2.1 gal) containers lined with polyethylene
bags were filled with cooling water.  One container was covered to ex-
clude sunlight.  No effort was made to keep temperatures constant.  Oxi-
dant residuals were determined by the amperometric titration method in
both containers over a period of approximately 60 min.

     Total oxidant residuals were determined in receiving waters by for-
ward amperometric titrations at all plants except Moss Landing.   In this
plant, samples were taken for later titration, using the back-titration
method for free oxidant residual.  This method was necessary because the
turbulence and wave action at the outfall caused the microammeter needle
of the titrator to vibrate and made the end-point difficult to read.
Receiving water samples were generally taken at the surface,  although
some samples were taken at depths of 1, 2, and 3m (3.3,  6.6,  and 9.9
ft).  Sampling point locations were determined by reference to onshore
features and in some cases by a rope attached to the outfall structure.

     Analytical Methods and Equipment

     To measure oxidant residuals, two methods are used, amperometric
titration and DPD-FAS titrimetric method.

1.   Amperometric titration.  Oxidant residuals were determined ampero-
     metrically using Fischer and Porter  (Model 17T1010) portable titra-
     tors.  Total oxidant residuals (TOR) were determined by a direct
     titration according to standard methods  (1974).  TOR could be
     determined in approximately 1.5 min by applying some suggestions by
     Manabe (1974).  These included the use of a graduated cylinder
     modified to measure a 200 m sample quickly and the addition of up
     to 90% of the titrant before turning on the stirrer.  The back-
     titration procedure described by White (1972) for the determination
     of free oxidant residual (FOR) was adopted because of an unstable
     end-point experienced during a direct titration.   The term
     "oxidant residual" has been used rather than "chlorine residual"
     because the amperometric method measures bromine and iodine resid-
     uals in addition^to chlorine residuals.
                                 196

-------
  EXHIBIT 9:  WATER QUALITY AND TIDAL DATA AT THE CONTRA COSTA POWER PLANT


Dal R/Sample
20 Oan 1976
#11 CONO-IM DUR
#14 COND-IN nUR
10 Feb 1976
#13 COND-IN BEF
#13 CONO-IN DUR
i?13 COND-IN AFT
16 Jim 1976
S'13 CO:-ID-IN BEF
//I 3 COMD-IN m.'R(-i-)'
/<13 COMD- IN DUR{-)**
' 7 OUT DUR
/,'13 COMD-IN AFT
14 Spt 19/6
#11 COND-IN PFF .
#12 corio-in nuR(+)
#11 COND-IN Dl'R(-)**
6 OUT DUR
#13 COND-IN AFT

NH3-N
niq/t

0.0-1*


0.11*
0.09*
0.10*

0.091
O.OG6
0.078
0.082
0.078


0.019
0.012
0.009
0.017

Orq-N
mq/l

0.13


0.29
0.21
0.20

0.47
0.10
0.43
0.11
0.36

0.33
0.31
0.33
0.34
0.32

Cl"
mg/0.

50
50

563
618
901

1130
1019


746

1032

913

761

Br"
nig/ 1




1.2
1.6
1.9

3.9
3.6
3.5
3.4
2.8

3.9

3.5

3.4

S.S.
mg/ 1,

22
19

42
30
35

111
118
128
114
80

34
37
34
32
41

pH









7.5-
7.7




7.7-
8.0




rime of
Study

0930-1200


0930-1120



1010-1100





1030-1250




Tidal Data


Time

Hi 0602
Lo 1213

Hi 1039
Lo 1841


Hi 0646
!.o 1415




Hi 0835
Lo 1403




Ht/ft

3.9
1.0

:1.0
0.?


3,8
0.2




3.0
I. 4



*  Determined by nesslerization.



   Samples taken during chlorination cycle from the condenser being chlorinated.



** Samples taken during chlorination cycle from a condenser not being chlorinated

-------
              EXHIBIT 10:   WATER QUALITY AND TIDAL DATA AT THE HUNTERS POINT AND POTRERO POWER PLANTS
<>
00

Date/Sample
Hunters Point
16 Dec 75
COUD-IN OUR
17 May 76
COND-IH BEF
CONO-fN DOR
OUT DUR
CON D- IN AFT
26 Aug 76
CONO-IN DEF
COMD-FN DUR
OUT DUR
COND-TN AFT
Potre ro
23 Feb 76
N. COMO-IN BEF
N. COND-IN DUR
OUT DUR
N. COMD-FN AFT
4 Mar 76
N COND-IN BEF
N CON D- IN DUR
OUT DUR
S COM D- IN AFT
NllrN
mg/t.


0.06*

0.134
0.047
0.036
n.119

0.128
0.037
0.024
0.140


0.055
0.000
0.032
0.075

0.118
0.059
0.077
0.104
Org-N
mg/e


0.33

0.18
0.08
0.03
0.14

0.34
0.28
0.28
0.36


0.27
0.20
0.21
0.26

0.20
0.16
0.21
0.17
Cl"
g/


15.4

18.2
17.7

17.9

18.7


18.4


16.8
16.5
16.5
16.4

16.7
16.8

16.8
Br~
mg/Jt.









72


73


67
71
70
71

63
64

66
S.S.
nig/K.


14

27
19
19
21

31
28
28
27


13
11
12
10





pH





7.0




7.5














Time of
Shudy


1100-1145

1040-1120




1040-1120





2100-2215




2100-2215



Tidal Data

Time




Hi 1655
Lo 09G9



Hi 1422
Lo 0813




Mi 2026
Lo 2509



Mi 2625
Lo 2000



Ht/ft




5.8
-1.1'



7.1
1.6




4.8
2.7



5.7
1.6


                Determined  by  nesslen'zation.

-------
EXHIBIT 11:  WATER QUALITY AND TIDAL DATA AT THE MOSS LANDING POWER PLANT
Date/Sample
12 Apr 76
7-1 COND-IN BEF
7-2 COND-IN DUR<+>*
7-1 COND-IN DUR(-)t
7-2 COND-IN AFT
19 Apr 76
7-1 COND-IN BEF
7-2 COND-IN DUR(-i-)*
7-1 COND-IN DUR (->''"
7 ISE DUR
7-2 COND-IN AFT
14 Jul 76
6-1 COND-IN BEF
6-2 COND-IN DUR(-t-)*
6-1 COND-IN DUR(-) '
6 ISE DUR
6-1 COND-IN AFT
22 Jul 76
6-1 COND-IN BEF
6-2 COND-IN DUR!+)*
6-1 COND-IN DUR!-r
6-1 COND-IN AFT
5 Aug 76
6-1 COND-IN BEF
6-1 COND-IN DUR<+)*
6-2 COND-IN DUR(-)t
6 ISE DUR
6-1 COND-IN AFT
23 Sept 76
6-1 COND-IN DEF
6-1 COND-IN DUR(t)*
0-2 COND-IN DUR(-)f
6 ISE DUR
6-1 COND-IN AFT
NH3-N
mg/i

0.096
0.021
0.114
0.137

0.058
0.005
0.033
0.016
0.037

0.046
O.COO
0.038
0.010
0.047

0.072
0.010
0.120
0.093

0.074
0.016
0.072
0.045
0.074

0.052
0.045
0.071
0.017
0.103
Org-N
mg/J

0.20
0.09
0.19
0.25

0.1*
0.08
0.12
0.10
0.04

0.19
0.19
0.19
0.19
C.I9

0.30
0.19
0.30
0.26

0.22
0.17
0.21
0.17
0.23

0.19
0.25
0.22
0.16
0.30
cr
g/n

19.0
18.7
18.8
18.5

18.9
19.0
18.8

19.1

18.6



19.1

18.8


18.9

18.8



19.1

18.7



18.6
Br"
mg/i

78
82
81
82

68
70
67

74

82



80

78


74

73



70

70



67
S.S.
rng/t.

19
22
21
21

31
14
IS
19
20

12
14
8
12
8

15
9
11
10

23
18
18
16
18

3
5
4
2
4
PH

7.9-
8.0



7.8





8.1-
8.2




8.0-
8.1



8.2-
8.3




8.9




Time of
Study

1240-
1330



1300-
1350




1300-
1400




1300-
1350



1300-
1400




1300-
1350



Tidal Data
Time

HI 0838
Lo 1438



Hi 1533
Lo 0840




Hi 1403
Lo 0720




Hi 0908
Lo 1327



Hi 0826
Lo 1307




Hi 1044
Lo 1656



Ht/ft

4.7
0.5



4.1
-0.6




4.8
-0.4




3.5
2.8



3.8
2.7




5.4
0.3



        *  Samples  taken  during  chlorination  cycle  from  the  condenser
        being chlorinated.

        f  Samples  taken  during  chlorination  cycle  but from  a  conden-
        ser not being chlorinated.

-------
2.   DPD-FAS titrimetric method.  The DPD-FAS titrimetric method of
     Palin  (1957, 1961, 1975) was used to distinguish chlorine and
     bromine residuals.  Three separate titrations give:  free chlorine
     plus free and combined bromine; combined chlorine; and free and
     combined bromine.  The separate determination of free and combined
     bromine is not possible.

     Organic nitrogen, chloride, bromide, and suspended solids were all
determined according to standard methods (1976).  Ammonia-nitrogen was
determined by the phenolhypochlorite method of Solorzano  (1969).  This
method was found to be well suited for fresh, marine, and estuarine
waters by Zadorojny et al.  (1973) .
RESULTS

     In this section, we describe the results of our in-plant analyses
and decay and recovery water analyses.

     In-Plant Studies

     Persistence and decay of oxidant residuals at the outfalls and in
the receiving waters can best be explained after the nature and dis-
appearance of the residual inside the plants are understood.  The in-
plant studies were conducted to establish the nature of the oxidant
residual and to gain an understanding of the demands in the cooling
water systems at each of the power plants.  Each plant was studied sep-
arately because of variations in water quality, chlorine dosage, fre-
quency and duration of chlorination, and flow time through the system.

     In-plant studies were conducted over a period of 1 year to be in-
clusive of varying water quality conditions.  Typical results at each
of the four plants studied are presented in Exhibits 12 to 15.  The
graphs show the temporal variation of oxidant residual during a chlori-
nation cycle.  Also shown is the chlorine dose calculated on the basis
of the chlorine injection rate determined by power plant personnel and
the cooling water flow rate.  The calculated dose may not be precise,
because the chlorine injection rate was difficult to read accurately.
In addition, the only value usually known for the cooling water flow
rate was the original design value.  

          Contra Costa Power Plant  The results of the in-plant field
studies conducted at the Contra Costa power plant are summarized in
Exhibit 16.  Water quality and tidal data corresponding to most of
these studies are presented in Exhibit 9.

     A combined residual of from 0.10 to 0.40 mg Cl / formed by the
time the cooling water reached the COND-IN  (approximately 1.2 min of
flow time)  and generally remained unchanged at the outfall when the
dilution effect was taken into account.  It appeared that the FOR which
                                200

-------
      1    I
      CHLORINE INJECTION RATE: 1600-1100 )bt/24hr>
      CHLORINE DOSE (CALCULATED]' 1.7-1.8 mj/l
      KEY;  CONO-IN OUT
                0  TOR (AMPEROMETRIC)
                D  FOR [AMPEROMETRIC)
       TWO TIMES MEASURED VALUES
       I   I    I   I    I   I    I    I	1	I	1
      1014  1018  1022 1026 1030 1034  1038  1042  1046 1090  1054
                   TIME

EXHIBIT 12:   IN-PLANT  STUDY  AT THE
   CONTRA COSTA  POWER PLANT,  6  JUNE
   1976,  UNIT  7,  CONDENSER NO.  13
      -i	1	1	1	1	1	1	1	r
       CHLORINE INJECTION RATE: ISOO lbi/24hn
       CHLORINE DOSE (CALCULATED): 2.5 rag/1
       KEY'  CONO-IN  OUT
                     TOR {AMPEROMETRIC)
                     FOR fAMPEROMETRIC!
                     FREE CL2+ BROMINES (DP01
                     COMBINED CL2 (OPD)
                     BROMINES IOPD)
   1040 1044  1048 1052  1056  1100  IIO4  1106  1112  1116  1120
                    TIME

EXHIBIT 13:   IN-PLANT  STUDY  AT
   THE HUNTERS  POINT  POWER PLANT,
   26  AUGUST  1976,  UNIT 4
                                                      o
                                                      I
                                                      J0.8
      CHLORINE INJECTION RATE: 1400 lt>s/24hn
      CHLORINE DOSE (CALCULATED): 1.6 mg/1
      KEY'  COND-IN OUT
               O   TOR (AMPEROMETRIC)
                                                                         -TOR AT COND-IN
                                                                               TOR AT OUT"
              TWO TIMES MEASURED VALUES
                   I    I   I	L_
  0900 0904 0908 0912 0916 0920 0924 0926 OB52 0936 0940 0944
                   TIME
   EXHIBIT  14;    IN-PLANT STUDY
     AT  THE PQTRERQ POWER PLANT,
     4  MARCH  1976,  UNIT  3,
     N.  CONDENSER
    1255 1300 1305 130 136 1320 1325 1330 1335 1340 |345 1360 1355
                    TIME
EXHIBIT 15:   IN-PLANT  STUDY
   AT  THE MOSS LANDING  POWER
   PLANT, 19  APRIL 1976,
   UNIT  7
                                           201

-------
to
o
                       EXHIBIT 16:  SUMMARY OF FIELD RESULTS AT CONTRA COSTA POWER PLANT
                                                   UNITS 6 and 7
Date
20 Jan 76
10 Feb 76
16 Jim 76
14 Sppt 76
C12
Dose
Calculatec
mgCl2/i!.
1.6
1.6
1.2
1.3
Condenser-Inlet
Amp. Tit.
TOR
mgCl2/'-
0.75
0.50
0.55
0.30
FOR
mgCl2/
0.45
0.10
0.36
0.20
DPD-FAS*
A
mqClf/l



0.20
6
mrjCl?y



0.10
C
nigCl2/f



0.20
Outfall Manhole"1"
Amp. Tit.
TOR
mgCl2 /
0.18
0.23
0.18
0.04
FOR
nigCl2/S
0.03
0.00
0.08
0.00
DPD-FAS*
A
mgCl2 /I



0.00
B
mgCl?,".



0.03
C
mgCl2 / '



0.00
                 *  Results of three titrations  are:   A  =  Free  chlorine  plus  bromine  residuals
                                                      B  =  Combined  chlorine residual
                                                      C  =  Bromine residuals.
                    Chlorinated  cooling  water  flow  was  diluted  with  an  approximately equal  volume of
                    unchlorinated  water  immediately after  the condenser.

-------
disappeared accounted for most of the loss of residual between the
COND-IN and the outfall.

     The DPD-FAS results for the study of 14 September 1975 indicated a
combined chlorine residual of 0.10 mg/ at the COND-IN and 0.03 mg/ at
the outfall, both of which compare favorable with the amperometric titra-
tion results.  The FOR of 0.20 mg C12/& determined amperometrically ti-
trates as bromine residual by the DPD-FAS method since titration  "A"
equals titration "C."

     Cooling water at Contra Costa varied from 0.3% to 6.1% sea water
during the dry summer months.  Most of the water quality data show the
effects of tidal changes.  The addition of chlorine generally caused a
decrease in ammonia-nitrogen concentration, but this, was more evident
at the other power plants.  Compared with earlier studies, the much
lower combined residual detected on 14 September 1976 is explained by
tV>e very low Nf^-N concentration.

          Hunters Point and Potrero Power Plants  The results of the in-
plant studies conducted at the Hunters Point and Potrero power plants
are summarized in Exhibit 17.  Water quality and tidal data correspond-
ing to these studies are presented in Exhibit 10.

     In general, FOR equaled TOR both at the COND-IN and at the outfall.
An exception was the study of 17 May 1976, which detected a combined
residual at the outfall of 0.13 mg Cl2/ that had formed during the
approximately 0.9 min of flow time from the COND-IN.

     The DPD-FAS results for the study of 26 August 1976 at Hunters
Point revealed a small combined chlorine residual at the COND-IN, which
the amperometric method failed to detect.  The results at the COND-IN
are difficult to explain since titration C should never be greater
that titration A.  At the outfall, a total residual of 0.45 mg Cl2/
was identified as bromine residual by the DPD-FAS method.

     A TOR of 1.0 Cl2/ was desired at the COND-IN for the studies of
23 February 1976 and 4 March 1976 at the Potrero power plant.  This
criterion was lowered to 0.5 mg/ and was the criterion during the
studies on October 15 and 17.  The studies of 4 March 1976 and 15 Octo-
ber 1976 revealed essentially no loss of oxidant residual between the
COND-IN and outfall manhole, which corresponded to a flow time of
approximately 0.4 min.

     Cooling water at Hunters Point and Potrero varied from 79% sea
water.  Ammonia-nitrogen concentrations determined on samples collected
at the COND-IN decreased by up to 0.091 mg/Jl during chlorination.  A
decrease in organic nitrogen was also evident during chlorination.

          Moss Landing Power Plant  The results of the in-plant studies
at the Moss Landing power plant are summarized in Exhibit 18.  Water
quality and tidal data are presented in Exhibit 11.
                                203

-------
to
o
                        EXHIBIT 17:  SUMMARY OF FIELD STUDY RESULTS AT THE HUNTERS  POINT
                                      (UNIT 4)  AND POTRERO POWER PLANTS (UNIT 3)
Date
Hunters Pt.
16 Dec 76
17 May 76
26 Auq 76
19 Och 76 H
Potrero
23 Fob 76
4 Mar 76
15 Oct 76
27 Oct 76
C12 Dose
Calculated
mgCl ?/f.
1.6
1.6
2.5
1.2
2.0
1.6
0.9
0.9
Condenser Inlet
Amp. Tit.
TOR
mgCl2/H
0.85
0.72
1.00
0.65
1.3-1.5
0.8-1.0
0.73
0.73
FOR
mgCl2/8.
0.72
1.00
0.65
DPD-FAS*
A
mgC!2/e,
0.60
B
mgCl2/J.
0.10
C
mgCl2/f,
0.90
Outfall Manholet
Amp. Tit.
TOR
mgCl2/S.
0.50
0.50
0.70
0.24
0.5-0.6
0.4-0.5
0.35
0.28
FOR
rngC12/H
0.37
0.70
0.24

DPD-FAS*
A
mgCl2/P.
0.45

B
mgCl2/f
o.do

C
mgCl2/fc
0.45

              *  Results of three titrations are:
A = Free chlorine plus bromine residuals
B = Combined chlorine residual
C - Bromine residuals.
                 Hunters Point - The cooling water flows through both condensers of Unit 4 and is
              chlorinated simultaneously.  Potrero - Chlorinated cooling water was diluted with an
              approximately equal volume of unchlorinated water immediately after the condensers.
              ** Cooling water flows through the condensers were chlorinated separately for this study.

-------
                      EXHIBIT 18:  SUMMARY OF FIELD STUDY RESULTS AT THE MOSS LANDING POWER PLANT

                                                   (UNITS 6 AND 7)
to
o
ui
Date
12 Apr 76
1? Apr 76
14 Jul 76
22 Jul 76
5 Aug 76
23 Sep 76
C12
Dose
Calculated
mgCl2/H
2.2
2.2
2.1
2.1
1.3
2.1
Condenser Inlet
Amp. Tit.
TOR
mgCl2/5.
1.35-
1.65
1.45-
2.10
1.40-
1.30
1.30-
2.10
0.80-
1.20
1.00-
1.70
FOR
mgd2/
-------
     The results generally indicate much higher oxidant residuals at
the COND-IN than the 1.0 mg/ desired by the plant chemist.  Measure-
ments by the amperometric and DPD-FAS methods were often on the order
of two times the value determined by the plant chemist using the ortho-
tolidine method.

     At the COND-IN, FOR was generally greater than or equal to TOR.
Since FOR can never be greater than TOR, an error (probably in the TOR
measurement) was indicated.  Approximately 1 min was required to measure
out a 200 m sample, transfer it to the titration jar, add the KI and
pH 4 buffer, and perform the TOR titration.  During this time, the oxi-
dant residual was reacting with species in the water and decaying.  On
the other hand, the back-titration method for FOR fixes the concentra-
tion as soon as the sample is mixed with excess phenylarsine oxide and
pH 7 buffer.  Another contributing factor to this result was the flash-
ing off of iodine during stirring in the forward titration for TOR.

     The DPD-FAS results indicated little or no combined chlorine resid-
ual at the COND-IN.  Some combined chlorine was indicated at the
sampling point at International Shellfish Enterprises, Inc., both by
the amperometric and DPD-FAS methods.  However, most of the DPD-FAS
results must be regarded with suspicion because of the discrepancy
between titrations A and C.

     Cooling water at Moss Landing varied from 95% to 99% sea water.
Ammonia-nitrogen concentrations determined on samples collected at the
COND-IN decreased by up to 0.075 mg/ because of chlorination.  A
decrease of organic nitrogen during chlorination was observed in four
of the six studies.

          Summary  The DPD-FAS titrimetric method was added to the field
studies to gain a better understanding of the nature of the oxidant
residual at the power plants.  Exhibit 19 presents the results of two
studies that were conducted in the laboratory to compare this method
with the amperometric method.  Chlorine (sodium hypochlorite)  was
added to 4 samples of San Francisco Bay water, and oxidant residuals
were determined for a period of 40 min.  A good correlation between FOR
determined amperometrically and titration A of the DPD-FAS method was
obtained.  TOR determined by the amperometric method was equal to or
slightly less than TOR calculated from the DPD-FAS results (titration A
plus B).   The discrepancy between amperometric and DPD TOR data appeared
to result from the underestimation of combined chlorine residual by the
amperometric method.  Results of titration C 'indicated that most of the
FOR was bromine residual.

     Exhibit 20 summarizes the disappearance of oxidant residual
observed for all the in-plant studies.  Loss of residual is expressed
as a percentage of the chlorine dose.  The table does not include the
decrease of oxidant residual because of dilution immediately after the
condensers.   The measured TOR at the outfalls has been doubled before
the percentage loss was calculated for those plants where such a
                                206

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EXHIBIT 19:   CHLORINATION STUDIES
  CONDUCTED  AT SERL COMPARING  THE
  DPD-FAS  TITRIMETRIC AND AMPERO-
  METRIC TITRATION METHODS
                                                                30 JUN 76
                                                          S. F. BAY WATER AT SERL
                                                          pH 7.6, CT I60g/l, NM--N 0.36mg/
                                                          CI DOSE 3.05M)/1, TEMP 21.0 "C
                                                               20
                                                            TIME, min
                                                                7 JUL 76
                                                          pH7.7, NH,-N 0.45mg/l
                                                          Cl DOSE 4.00 mg/l, TEMPI^O'C

                                                          KEY: , A  TOR (DPO-calculated)
                                                                TOH (AMPEROMETRIC)
                                                                FOR (AMPEROMETRIC)
                                                              A  FREE CI2+BROMINE (OPD)
                                                              B  COMBINED Clj (OPCi
                                                              C  BROMINES (OPO)
10      20
    TIME, min
        EXHIBIT 20:   SUMMARY OF LOSS  OF OXIDANT  RESIDUAL WITHIN
                    THE POWER PLANT COOLING WATER  SYSTEM

Power- Plant


Contra Costa

Hunters Point
Potrero
Moss Landing

Unit


6
7
4
3
6,7
Approximate
Flow Time (ir.in)
IfiJ. PT.
To
COND-IN
1.2
1.2
1.5
0.9
0.3
COND-IN
To b
OUTFALL
1.5
0.9
0.9
0.4
6-7

Total

2.7
2.1
2.4
1.3
7-8
TOR LOSS (%)*
!f!J. PT
To
COMO- IN
55-75
54-67
44-59
19-44
10-32
COND-IN
To
Outfall*
3-21
6-17
10-34
0-19
41-64
TOR (%)*
Remaining
At OutfallT

17-24
27-31
24-40
54-73
16-27
     *   Percent  loss  does  not include  that caused by dilution
     immediately past the  condensers.
         Sampling part at Moss Landing.
                                    207

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dilution occurred.   Flow times between the sampling points at  each plant
are indicated.   Results  at Contra Costa are separated for Units  6  and 7
because of the different flow times from the COND-IN to the outfall.

     Exhibit  20  shows  that the disappearance of TOR generally  depends on
the flow time through  the system.  This is especially evident  from the
COND-IN to the outfall because of the wide variation of flow times that
existed.  Exhibit  21 is  a graph of the percentage loss of TOR  between
the COND-IN and  the  outfall sampling point as a function of flow time.
EXHIBIT 21:  PERCENTAGE  LOSS OF
  TOR BETWEEN THE  COND-IN AND THE
  OUTFALL SAMPLING POINT
                                                        KEY

                                                    POTRERO P. P. - UNIT 3

                                                   D CONTRA COSTA PP. - UNIT 7

                                                    CONTRA COSTA P. P. - UNIT 6

                                                   A HUNTERS POINT P. P. - UNIT 4

                                                   O MOSS LANDING P. P. - UNITS 6 & 7
234567
  FLOW TIME - COND-IN TO OUT, min
     Decay and Receiving  Water Studies

     Decay studies were conducted on the chlorinated cooling water col-
lected at the outfalls to determine the approximate life of oxidant re-
siduals in the receiving  waters.   Oxidant residuals were measured at  var-
ious locations in the receiving waters to determine the extent of the
area affected by the power plant  discharge during chlcrination.

     The results of the decay  studies conducted at the outfalls are
presented in Exhibits 22-25.   The loss of oxidant residual between the
point of chlorine injection, the  COND-IN, and the outfall are shown.
The effect of the 1:1 dilution, which occurred between the COND-IN and
the outfall sampling point,  is indicated by a dotted line for all plants
                                 208

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EXHIBIT 22:  RESULTS OF DECAY
  STUDIES CONDUCTED AT THE CONTRA
  COSTA POWER PLANT, UNITS 6 AND 7
  OUTFALL
                                      EXHIBIT 24:   RESULTS OF
                                        DECAY STUDIES CONDUCTED
                                        AT  THE POTRERO POWER
                                        PLANT, UNIT 3 OUTFALL
I
O 0.2
-TOR (DARK)
   (LIGHT)
EXHIBIT 23:   RESULTS OF DECAY
  STUDIES CONDUCTED AT THE HUNTERS
  POINT POWER PLANT, UNIT 4 OUTFALL
                                                        TOR (DARK)
                                                         /TOR (LIGHT)
                                                   0  20  30 40  50 GO  70
                                                       TIME, min
                                     EXHIBIT 25:   RESULTS OF DECAY
                                       STUDIES CONDUCTED AT THE
                                       MOSS LANDING  POWER PLANT,
                                       UNITS 6 AND 7 AT SAMPLING
                                       POINT AT SHELL ENTERPRISES
                                 209

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except Hunters Point, where flows to both condensers were chlorinated
simultaneously.  Exhibit 26 is a summary of the decay studies.  It
should be remembered that the results are based on field studies, and
that no attempt was made to -keep temperature constant.  In most cases,
temperature increased in the "light" experiments and decreased in the
"dark" experiments.

     The results of receiving water studies are presented in Exhibits
27-31 in the form of isoconcentration lines of TOR at the surface.  Sur-
face TOR was always found to be the highest compared to samples taken at
various depths.  Whenever possible, the 0.02, 0.05, and 0.10 mg C12/&
isoconcentration lines are shown together with the enclosed surface
areas.  These areas give an approximation of the regions affected by
various levels of TOR.

     The results of Contra Costa (Exhibit 27) show the effects of chlor-
ine addition to one condenser of Unit 6.  The study of 8 July 1976 was
conducted during a flood tide so that no residual was detected down-
stream from the point where the discharge canal meets the river.  The
study of 3 December 1976 shows the effect of an ebb tide.  A TOR of
0.02 mg C12A was detected some 150 m (500 ft) down the San Joaquin
River near the intake structure for Units 6 and 7.  The plume boundary
was clearly'visible and was confirmed by both oxidant residual measure-
ment and temperature readings.

     Exhibit 28 presents chlorine addition to one condenser of Unit 4
at Hunters Point.  A TOR of 0.02 mg C12/-& was measured up to 150 m
(500 ft) from the outfall structure.  The dilution effect of varying
tidal stages is evident.  The studies of 19 October 1976 and 3 November
1976 were conducted at lower tides than the other two studies.  Tidal
effects are very pronounced because the discharge structure is sub-
merged at high tides but not at low tides.

     Results at Potrero are presented in Exhibits 29 and 30.  Exhibit
29 shows the effects of chlorine addition to one condenser of Unit 3.
A TOR of 0.02 mg Cl2/fc was detected up to 168 m (550 ft) from the out-
fall structure and was drawn into the No. 1 and No. 2 unit intake.
Tidal effects were not as evident as at Hunters Point because the
Potrero outfall structure is never fully submerged.  Exhibit 30 shows
the effects of chlorine addition to all condensers of Units 1 and 2.
These units are very old, and chlorination of each condenser separately
is not possible.  A TOR of 0.02 mg C12/& was detected up to 400 m
(1300 ft)  from the outfall structure.

     A study was conducted in the receiving water at Moss Landing on
23 September 1976 (see Exhibit 3.1) .  It was very difficult to keep the
boat in the discharge plume in Monterey Bay because of the force of
37.9 m3/sec (600 000 gpm) of cooling water reaching the surface.  It
was difficult at times to determine the exact location of the plume
because of waves continually moving the boat.  A maximum TOR of
                                210

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EXHIBIT 26:  SUMMARY OF DECAY STUDIES

Date



10 Feb 76

16 Jun 76

17 May 76

26 Aug 76

23 Feb 76
4 Mar 76
19 Apr 76
14 Jul 76


Power Plant



Contra Costa

Contra Costa

Hunters Point

Hunters Point

Potrero
Potrero
Moss Landing
Moss Landing


Initial
Temp.
ro




34.5

24.0

30.0

20.0
20.0
24.0
27.0


Initial
TOR
(mgCl2/*)


0.23

0.16

0.52

0.72

0.61
0.43
0.22
0.20


"Dark"
or
"Light"


Dark
Light
Dark
Light
Dark
Light
Dark
Light
Dark
Dark
Light
Dark
Light
TOR After
30 min


mgC12/.
0.14
0.12
0.09
0.07
0.10
0.03
0.20
0.72
0.20
0.15
0.08
0.08
0.05
% of
Initial
TOR
61
52
56
44
17
6
28
17
33
35
35
40
25
60 min


mgCl2/)l
0.12
0.09
0.08
0.06
	
--
0.15
0.08
0.13
0.10
0.03
0.05
0.02
% of
Initial
TOR
52
39
50
38


21
11
21
23
13
25
10
Time to Reach
0.1 mgCl2/
(min)



> 90
45
20
8
30
10

38
> 90
60
22
16
8

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       \
                                8 JUL 76
                            HI WATER- 1447
                            LO WATER - 0837
                            STUDY- 1025-1050
                SAN JOAOUIN RIVER
                                    3 DEC 76
                                 HI WftTER-1259
                                 LO WATER-2033   ]
                                 STUDY-1550-IG25
         PLUME BOUNDARY
0.00   00!   002    002   002   002


                 ~*^77mn2D	~\ 0 03
                            117 Zl


              mUJ(^'"i"   I   0.04

             UNIT .-Til	I;;"'  " "i
                   '- '""""\	}

                (TTMP.-C - IN PARENTHESES)
     UNIT 6
     COIJD-IN
     0.50
EXHIBIT  27:    TOR  (mgClg/Jl)  AT  THE SURFACE
   OF THE RECEIVING WATER AT  THE CONTRA
   COSTA  POWER PLANT,  UNITS 6 AND  7
                     19 OCT 76
                   HI WATER 0959
                   LO WATER 1&57
                   STUDY 1320-1410
                 1260OO FTZ
                                           5 NOV 76
                                          HI WATER 0941
                                          LO*ATER 1617
                                          STUDY I4IO-I5OO
                     17 NOV 76
                   HI WATER 0811
                   LO WATER 1445
                   STUDY LL30-IZ1O
                I560O FT2
                                            23 NOV 76
                                          HI WATER 1228
                                          LO WATER I94O
                                          STUDY 1320-I35O
                                            SCALE: | IN. a 200 FT
EXHIBIT 28:   TOR  (mgC!2/)  AT THE  SURFACE
   OF  THE  RECEIVING  WATER AT  THE HUNTERS
   POINT POWER  PLANT,  UNIT  4
                    212

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     OUT
    "-0.24
  17 NOV 76
HI WATER 0810
L.O WATER 1442
STUDY 0945-1030
                                              27 OCT T6
                                             HI WATER 1535
                                             LO WATER 0939
                                             STUDY 1350-1460
                                               19 NOV 76
                                             HI WATER 0929
                                             LO WATER 1621
                                             STUDY 0930-1012

                                           SCALE' I INi= 200PT
EXHIBIT  29:    TOR (mgCl2/)  AT THE  SURFACE
   OF  THE RECEIVING  WATER  AT  THE  POTRERO
   POWER  PLANT,  UNIT 3
    15 OCT 76
 Hi WATER 1711
 LO WATER 1135
 STUDY 1400-1530
            SCALE'  I IN. = 300 FT
            27 OCT 76
          HI WATER 1533
          LO WATER 0939
          STUDY 1130-1200
                                             COND-IN -0.89
EXHIBIT  30:   TOR  (mgC!2/Jl)  AT THE SURFACE
   OF  THE RECEIVING WATER AT THE  POTRERO
   POWER  PLANT,  UNITS 1  AND 2
                    213

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 EXHIBIT 31:  FOR (mgCl2/)
   AT THE SURFACE OF THE
   RECEIVING WATER AT THE
   MOSS LANDING POWER PLANT,
   UNIT 6
                                     t
OCEAN
WAVES
                                             0.12
                                                  UNIT 6
                                                          0.16
                                       0.00
                                                   STRUCTURES"'
                                                   0.12
                                                                 0.06
                                                                  +
                                                                 0.09
                                             UNIT 7
                                   MONTEREY

                                    BAY
                                                  0.00
    SCALE' I IN. a 20 FT
0.16 mg C12/& was determined at the surface at a time when the TOR
ranged from 1.2 to  1.7 mg/  at the COND-IN and from 0.31 to 0.33 mg/
at the sampling point at  International Shellfish Enterprises, Inc.
DISCUSSION OF" RESULTS

     In this section, we  describe the nature of the oxidant residual,
chlorination practices, demands in cooling water system, decay and
receiving water studies.

     Nature of the Oxidant Residual

     The nature of the oxidant residual at each of the power plants  is
important for understanding the decay and persistence in the receiving
waters.  None of the methods currently available for the routine mea-
surement of oxidant residuals unequivocally identify the compounds that
compose the residual.  This is especially true for oxidant residuals
in estuarine and marine waters.

     When chlorine is added to sea water that is free of amino-nitrogen,
bromide (present at a concentration of approximately 70 mg/) is oxi-
dized to bromine as follows:

     HOC1 + Br~ = HOBr +  Cl~.
                                214

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     Exhibit 32 shows the time required  (based on  reported rate con-
stants) to convert 99% of the chlorine to  bromine  at a temperature of
25C and a pH of 8.3 for varying percentages  of sea  water (1976).   The
reaction proceeds to 99% completion within 10 sec  in high salinity
water, but very little bromine is formed in water  containing less  than
3% sea water.
 EXHIBIT  32:  TIME  REQUIRED
   FOR 99%  CONVERSION  OF  FREE
   CHLORINE TO HOBr AT 20C
   AND A  pH OF 8.3
10     20
 % SEA WATER
     A similar oxidation of iodide occurs.  Total oxidation to iodine
would account for a 0.035 mg/H residual as chlorine.

     When ammonia-nitrogen is present in sea water, there is a competi-
tion between the ammonia and the bromide ion for chlorine.  A number of
reactions are possible, depending on pH, chlorine dose, concentration
of NH^-N, salinity, and reaction time.  The formation of chloramines is
generally favored by high OT^-N, low salinity, high pH, and low chlorine
dosage.

     Using an equilibrium model, Sugam and Helz (1976) suggest that free
bromine and bromamines predominate over the chlorine species at salin-
ities above approximately 0.3 g/S, with ammonia concentrations "typical"
of estuarine and marine waters.  The predominant species in sea water
containing 0.08 mg NH3-N/& dosed at 1.0 mg C12/& at pH 8.1 were tribro-
mamine (NBr3)  and dibromamine (NHBr2)  Equal amounts of monochloramine
(NH2C1) and HOBr were estimated to exist in a typical sea water of 35 g/
                                215

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salinity at pH 8 containing 0.12 mg/fc NH-j-N dosed at 0.5 mg C12A.  The
model for 0.3 g/H salinity is very complicated showing both chlorine and
bromine compounds with no two species in abundance between pH 6 and 8.

     Sollo e_t al. (1971) performed studies on the resulting speciation
when solutions containing both ammonia and bromide were dosed with
chlorine and residuals were determined after 2 min.   Solutions con-
taining 0.41 mg/ NH3-N and bromide concentrations of 3 and 25 mg/
dosed at 2 mg C!2/ are interesting,  because they most closely model
the conditions existing in cooling waters studied in this investigation.
Very little free bromine existed over the entire pH range tested (4.3 to
9.0).  At pH 7.4 and 25 mg/ Br~, a total halogen concentration of
2.8 mg Br2/& was detected, which was significantly lower than the 4.5 mg
Br2/ expected.  The entire residual was determined to be combined bro-
mine and was believed to be NHBr2/ whose instability might explain the
drop in total halogen residual.  At pH 7.4 and 3 mg/ Br , the entire
4.5 mg Br2/& total halogen residual was determined to be chlorine
residual.
     Exhibit 33, after Johnson and Overby (1971) , shows the bromirte
species postulated to be present after 1 to 2 min as a function of both
pH and the logarithm of the initial mole ratio of ammonia to bromine.
These data 'suggest that HOBr and NBr3 would predominate after 1 to 2
min under conditions that exist at the Hunters Point, Potrero, and Moss
Landing power plants if a total conversion of chlorine to bromine is
assumed.  NHBr2 would be predominant at somewhat higher ammonia con-
centrations than were measured at the power plants.   At Contra Costa,
both chlorine and bromine compounds are suggested with prediction of
individual species being difficult.
                         I     I     I    1     F    1
                                                     10
EXHIBIT 33:  PRINCIPAL SPECIES OF BROMINE AND BROMAMINE PREDOMINATING
  AFTER 1-2 MIN AT VARIOUS pH AND AMMONIA TO BROMINE RATIOS.   (Lines
  represent equal equivalent concentrations.  Hypobromous acid  separa-
  tion from bromine given for 10"^ M bromide, after Johnson and Overby,
  1971.)
                                 216

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     It is apparent that both chlorine and bromine residuals generally
exist when chlorine is added to estuarine and marine waters.  Both the
amperometric and DPD-FAS methods were used to determine oxidant resid-
uals.  The amperometric titration method for total residual determines
all oxidant residuals whether they be chlorine, bromine, or iodine.  It
is apparent from Exhibit 19 that the amperometric free residual deter-
mination measures both free chlorine and bromine residual.  Since the
DPD-FAS method does not distinguish between free and combined bromine
residual, it is possible that some combined bromine existed and was
picked up by the amperometric method as "free residual."  This has been
suggested by Johannesson (1960), Palin (1961), and Sugam and Helz (1976).
Therefore, the so-called "free chlorine" residual measured by the stan-
dard amperometric method in estuarine or marine water is neither totally
"chlorine" nor totally "free."

     Chlorination Practices

     Chlorine dosage at many of the power plants was decreased during
the course of this study.  This was done both as a reaction to an aware-
ness of greater than expected residuals and to decrease the residual
in the discharge stream.  Those plants using the orthotolidine method
to set the chlorine dose were often chlorinating at twice the desired
level because of the inaccuracies of the analytical method.

     Total chlorine use per day for the four power plants located on the
San Francisco Bay system averages about 0.30 metric tons/day  (0.33 tons/
day) based on 1971 figures  (1973).  This represents 1.25% of the total
average daily use of chlorine of 34 metric tons/day  (37.5 tons/day) for
the  1975-1976 season as determined by Russell and Home  (1977).

     Demands in CoolingWater System

     Exhibits 20 and 21 indicate that the loss of oxidant residual in
the  cooling water system was a  function of flow time.  The longer the
cooling water was held within the plant, the lower the oxidant residual
that reached the receiving water.  This loss of residual was a result of
many factors, including reaction with ammonia-nitrogen, demand by inor-
ganic reducing agents, reaction with organic nitrogen, and uptake by
organisms living in the cooling water .system.  It is difficult to say
which of these are most important.  Exhibits 20 and 21 suggest the 'flow
time through the cooling water  system may be a good method to estimate
the oxidant residual at various locations with varying doses of chlo-
rine  .

     Decay and Receiving Water  Studies

     The decay studies  (Exhibits 22-26) show that the most persistent
residual existed at the Contra  Costa power plant.  From  50% to 52% of
the  0.16 and 0.23 mg/ TOR determined at the outfall remained after
60 min in those samples covered to exclude sunlight.  From 38% to 59% of
the  0.16 and 0.23 mg/SL TOR lasted for the same time period in samples
                                217

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exposed  to  sunlight.  This persistence is  likely explained by  a  slowly
decaying combined  chlorine residual that existed at Contra Costa.

     The highest concentrations of TOR remaining after  30 and  60 min
were at  Hunters Point and Potrero.  However, this was mostly a result of
the greater initial  concentrations of 0.43 and 0.72 mg/& TOR.  The  low-
est concentrations of TOR in decay studies were found at Moss  Landing,
where  the long flow  time to the outfall sample point produced  small ini-
tial TOR of 0.22 and 0.20 mg C12/&-  These were approximately  the same
initial  TOR that existed at the Contra Costa outfall; however, the  oxi-
dant residual at Moss Landing decayed to much lower values.  After  60
min at Moss Landing  from 0.02 to 0.05 mg Cl2/ remained, while at Contra
Costa  from  0.06 to 0.12 mg/ remained.  The major difference between  the
oxidant  residuals  is the predominance of bromine residuals at  Moss  Land-
ing, which  decay more rapidly than chlorine residuals (Sollo et  al.,
1971;  Johnson and  Overly, 1971; Johannesson, 1960).

     The presence  of sunlight appeared to speed up the rate of decay  at
all plants.  This  effect was most noticeable during the initial  20 min
decay  at Contra Costa on 16 June 1976 when a FOR existed.  After 20 min,
only combined residuals existed, and the effect of sunlight on decay  was
less pronounced.

     The significance of the oxidant residuals can be assessed in two
ways.  The  first approach is to consider the effect on an organism  such
as a fish that is  located at the discharge initially and then  swims
with a parcel of water away from the plume.  The second method is to
consider the effect  on an organism that remains in one spot and  is  sub-
jected to intermittent doses of chlorine.  Such a case would be  repre-
sentative of benthos or of a fish that remained stationary in  the dis-
charge.

     The first approach has been addressed by Mattice and Zittel (1976).
Exhibit  34, which  is derived from their data, shows several dose-exposure
time combinations  that result in acute toxicity;  the chronic toxicity
threshold is also  indicated for marine and fresh water organisms.  At
Contra Costa, the  cooling water discharge is into a canal that has an
approximate residence time of 12 min.  In the canal, average TOR con-
centrations were on  the order of 0.05 mg C12/^.   According to Mattice
and Zittel, this combination of chlorine dose and exposure time would
not result  in acute  toxicity to fresh water organisms.  However, it is
close  to Mattice and Zittel's acutely toxic level for marine organisms.
A TOR  of 0.02 mg C12A was detected for a distance of some 150 m
(500 ft)  down the San Joaquin River.  Assuming an average water veloc-
ity of 0.55 m/sec  (1.8 ft/sec), this results in an exposure time of
4.6 min  at  this TOR  concentration.  This TOR level is at Mattice and
Zittel's  chronic toxicity level for marine organisms.  For fresh water
organisms, the dose/exposure time stated to be acutely toxic to  fresh
water organisms by Mattice and Zittel is 0.02 mg/ for 200 min.
                                218

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         EXHIBIT  34:   ACUTE AND CHRONIC DOSES OF CHLORINE
                TO MARINE AND FRESH WATER ORGANISMS
                    (Mattice and Zittel, 1976)
            Marine Organisms       Fresh Water Organisms
TOR
(mgC!2/)
0.20
0.15
0.10
0.05
0.02*
Duration
(min)
0.50
1.0
2.5
13.0

TOR
(mgC!2/)
0.10
0.05
0.02
0.01
0.0015*
Duration
(min)
25
60
200
550

             Chronic toxicity threshold.
     Current velocities in the discharge plumes have been determined by
PG&E at the Hunters Point and Potrero Plants.  At Hunters Point, the
time to reach the 0.02 mg/Jl TOR isoconcentration line is 45 sec.
Assuming that the velocity, is constant between the outfall and the
0.02 mg/ TOR isoconcentration line, the estimated time to reach 0.05
mg/Jl TOR is between 11-28 sec; with the same assumption, the time to
reach the 0.10 mg/il TOR isoconcentration line is 7 sec.  These dose/
exposure times are far below the values quoted by Mattice and Zittel
to be acutely toxic to both fresh water and marine organisms.  Concen-
trations of TOR exist in the receiving water that are above the chronic
toxicity threshold for both fresh water and marine organisms.

     At Potrero, with an average discharge concentration of 0.3 mg/Jl
TOR, the time to the 0.02 mg/Jl TOR  isoconcentration line is approxi-
mately 3 min.  Using the same assumptions as for Hunters Point, the
time to the 0.1 mg/Jl TOR isoconcentration line is estimated to be be-
tween 48 and 56 sec; from the 0.1 mg/ TOR to the 0.05 mg/Jl TOR iso-
concentration line is between 20 and 65 sec.  The exposure between
the 0.3 mg/ TOR discharge and the  0.1 mg/Jl TOR isoconcentration line
(assumed to be an average of 0.2 mg/Jl TOR) is predicted to be acutely
toxic to marine organisms.  Acute toxicity is not predicted according
to the Mattice and Zittel data beyond this area.  Chronic toxicity to
marine organisms is predicted to exist out to the 0.02 mg/Jl isoconcen-
tration line and considerably further for fresh water organisms.

     The 0.16 mg Cl2/& maximum concentration measured at the surface of
Monterey Bay at the outfall from the No. 6 and No. 7 units at Moss
Landing could only have existed for a flow time of a few seconds from
the discharge pipes located at a depth of 6.1 m (20 ft).  At a distance
                                219

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of some 15.3 m  (50 ft) from the discharge, a residual of 0.09 mg C12A
was detected.  For this residual to be acutely toxic, according to
Mattice and Zittel's criterion, an exposure time of some 2.5 min must
exist.  It is questionable whether such a contact time existed between
the discharge structure and a point some 50 ft away on the surface.
These chlorine levels are above those suggested by Mattice and Zittel
as chronic toxicity threshold values for both fresh water and marine
organisms.

     The second approach has been investigated by Dickson e_t al^.  (1977),
who studied the effects of intermittent chlorination on goldfish and on
protozoans obtained from attached growths.  LC5Q values for goldfish
were found to be a function of total exposure time during a 24 hr
period.  In the temperature range of 17-22.5C, 2 hr exposure/day
resulted in an ,LC5Q of 1.18 mg/ TOR; with 3 hr exposure the LC5Q was
0.71 mg/i TOR; and with 4 hr/day exposure the LC5Q fell to 0.63 mg/
TOR.  These authors concluded that two to three exposures of 15-30
min/day at a TOR of 0.5-0.75 mg/X, would not result in lethality to the
goldfish.  Concentrations in the ranges stated above were never detected
in the receiving waters at any location sampled during this study.
Indeed, these levels were detected in outfalls only in early studies at
Hunters Point and Potrero before the reduction in chlorine dosage by
Pacific Gas and Electric.  Therefore, it might be concluded that with
current chlorination practice no lethality would result from chlorine
toxicity in cooling water to fish with the'same response as goldfish.

     The Cfekson et al.  (1977)  analysis on chlorine toxicity to benthic
protozoa was conducted at high (>0.58 mg/)  free chlorine concentrations
over a short period (2 hr) and cannot be compared directly with the
results obtained in this study.  Suffice it to say that these workers
noted significant decreases in numbers of individuals at all levels of
free chlorine tested.   Numbers of species and diversity were not
affected below 1.45 mg/Ji free chlorine dosed three times in 2 hr;
there was a decrease in number of species when the dosing was increased
to seven times in 2 hr at a level of 0.66 mg/&.  In general, it was
noted that chlorine exposure generally caused a shift in population
toward more tolerant species.
CONCLUSIONS

     We itemize our conclusions below.

     Chlorination Practice

1.   Chlorination frequency varied among the five power plants studied
     from I/week to 4/day.  Duration of chlorination varied from 15
     to 40 min per cycle.
                                 220

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2.    Oxidant residual determined at the condenser inlets was the crite-
     rion used by the power plants to determine the level of chlorine
     dose.

3.    During this study, the chlorine dose to produce the desired resid-
     ual of approximately 0.5 mgCl2/& at the condenser inlet was
     reduced.  This reduction resulted from a more precise measurement
     of chlorine residuals throughout the cooling system by our use of
     the amperometric method as compared to the orthotolidine method
     routinely utilized by power plant personnel,,

4.    All of the plants are currently using or are in the process of
     obtaining amperometric titrators because of the more stringent
     regulations that require accurate measurement of oxidant residuals.

     In-Plant Studies

1.    Total chlorine usage per day for the four power plants located on
     San Francisco Bay averaged about 0.3 metric tons/day (0.33 tons/
     day) based on 1971 figures.  This represented 1.25% of the total
     average daily use of 34 metric tons/day (37.5 tons/day) estimated
     for 1975-1976.

2.    Oxidant residuals determined at the condenser inlets were often
     significantly higher than the 0.5 or 1.0 mg/& desired by plant
     personnel, especially at those plants that use the orthotolidine
     method for control of chlorine residual.

3.    At Contra Costa, where the cooling water was the freshest, a com-
     bined residual of from 0.10 to 0.40 mgC!2/^ existed from the
     condenser inlet to the outfall.

4.    At Hunters Point, Potrero, and Moss Landing where cooling water
     ranged from 79% to 99% sea water, total oxidant residual generally
     equaled free oxidant residual at the condenser inlets.   Some
     combined residual was detected at the outfall of Hunters Point
     and at a sample point 6-7 min of flow time from the condenser
     inlet at Moss Landing.

5.    The DPD-FAS results indicated that most of the oxidant residual at
     Hunters Point and Moss Landing was bromine residual.

6.    The disappearance of total oxidant residual through the cooling
     water system generally depended on the flow time.  At Moss Landing,
     where the total flow time in the system was 6 to 7 min, from 16% to
     27% of the oxidant input remained at the outfall.  At Potrero,
     where the total flow time in the system was only 1.3 min, from 54%
     to 78% of the oxidant input remained at the outfall.
                                221

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     Decay and Receiving Water Studies

1.   The slowest decay at the outfall was observed at Contra Costa,
     where 50% to  52% of the total oxidant residual of  0.18 -  0.23 mg/l
     measured at the outfall remained after 60 min in these samples  that
     were covered  to exclude sunlight.  For the samples exposed  to sun-
     light,  38% to 39% remained  after the same time period.

2.   The most rapid decay at the outfall was observed at Hunters Point,
     where 17% to  28% of the total oxidant residual of  0.52 -  0.72 mg/&
     measured at the outfall remained after 30 min in the samples
     covered to exclude sunlight.  For the samples exposed to  sunlight,
     only 6% to 17% remained after the same time period.

3.   Maximum total oxidant residuals were found to exist at the  surface
     of receiving  waters.

4.   During  the chlorination cycle at Contra Costa, a 0.02 mgCl2/&
     residual was  measured some  500 ft from the point where the  dis-
     charge  canal  meets the San  Joaquin River.
 5.    During  the chlorination cycle at Hunters Point, 0.02
      residual was measured up to 500 ft from the No*. 4 outfall and at
      up  to 550 ft away from the No. 3 outfall at Potrero, and at up
      to  1300 ft from the No. 1 and No. 2 outfall at Potrero.

 6.    A maximum residual of 0.16 mgC!2/)l was measured at the surface of
      Monterey Bay above the outfall from the No. 6 and No. 7 units at
      Moss Landing.

 7.    Using the approach of Mattice and Zittel, it was determined that
      all effluents produced receiving water oxidant residual levels
      that would be predicted to demonstrate chronic toxicity to marine
      organisms; the receiving water from two plants  (Hunters Point and
      Moss Landing) showed levels that would be predicted to demonstrate
      chronic toxicity to fresh water organisms.  Acutely toxic levels
      to  fresh water and marine organisms existed in the receiving
      waters  at the Potrero site.
REFERENCES
American Public Health Association,  1976.  Standard Methods for the
  Examination of Water and Wastewater, 14th EdLNew York."~

Basch, R.E. and J.G. Truchan.  1976.  Toxicity of Chlorinated Power
  Plant Condenser Cooling Waters to Fiih^Environmental Protection
  Agency.  Duluth, Minnesota, publication number 600/3-76-009, 105 pp.

Brungs, W.A.  1973.  "Effects of Residual Chlorine on Aquatic Life,"
  Water Poll. Control Fed.  45:2180-2193.
                                222

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California Regional Quality Control Board.  1976.  San Francisco Bay
  Region, Waste Discharge Requirements For;  Pacific Gas and Electric
  Company-Hunters Point Power Plant, Potrero Power Plant, Orders No.
  76-60, 76-61, NPDES No. CA0005649, CA0005657, Oakland, California.

California Regional Water Quality Control Board.  1976.  Central Coast
  Region, Waste Discharge Requirements for Pacific Gas and Electric
  Company, Moss Landing Fossil Fuel Power Plant, Units 1-7, Monterey
  County, Order Ho. 76-09, HPDES Mo. CA0006254, San Luis Obispo,
  California.

Dickson, K.L., J. Cairns, Jr., B.C. Gregg, D.I. Messenger, J.L. Plafkin,
  and W.H. vander Schalie.  1977.  "Effects of Intermittent Chlorination
  on Aquatic Organisms and Communities."  Water Poll. Control Fed.  49:
  35-44.

Federal Register.  1974.  39(196).

Johannesson, J.K.  1960.  "The Bromination of Swimming Pools."  American
  Journal of Public Health, 5: 1731-1736.

Johnson, J.D. and R. Overby.  1971.  "Bromine and Bromamine Chemistry."
  J. San Eng. Div., Proc. Amer. Soc. Civil Engrs.  SA5:617-628.

Manabe, R.M.  1974.  Measurement of Residual Chlorine Levels in Cooling
  Water - Amperometric Method.  Environmental Protection Agency.  Cor-
  vallis, Oregon, publication number 660/2-73-039, 45 pp.

Mattice, J.S. and H.E. Zittel.  1976.  "Site-Specific Evaluation of Power
  Plant Chlorination."  Water Poll. Control Fed.  48:2284-2308.

Pacific Gas and Electric Company.  1973.  An Evaluation of the Effect
  of Cooling Water Discharges on the Beneficial Uses_pf Receiving
  Waters at;  Contra Costa Power Plant, Pittsburg Power Plant, Hunters
  Point Power Plant, Potrero Power Plant, San Francisco, California.

Palin, A.T.  1957.  "The Determination of Free and Combined Chlorine in
  Water by the Use of Diethyl-p-phenylene Diamine."  American Water^
  Works Association.  49:873-880.

Palin, A.T.  1961.  "The Determination of Free Residual Bromine in
  Water.  Water and Sewage Works."  American Water Works Association.
  108:461-462.

Palin, A.T.  1975.  "Current DPD Methods for Residual Halogen Compounds
  and Ozone in Water."  American Water Works Association.  67:32-33.

Russell, P.P. and A.J. Home.  1977.  The Relationship of Wastewater
  Chlorination Activity to Dungeness Crab Landings in the San Fran-
  cisco Bay Area.  Sanitary Engineering Research Laboratory.  Berkeley,
  California.  UCB/SERL Report No. 77-1, 37 pp.

                                223

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Selleck, R.E., A.O.K. Laird, and H.H. Sephton.  1976.  Optimization of
  Chlorine Application Procedures and Evaluatioii of Chlorine Monitoring
  Techniques.  University of California.   (Proposal to Electric Power
  Research Institute, Inc. Palo Alto.  March 25, 1976.)  Publication
  Number UCB-Eng-4180, 80 pp.

Sollo, F.W., T.E. Larson, and F.F. McGurk.  1971.   "Colorimetric Meth-
  ods for Bromine."  Environ. Sci. and Techn.  _5_: 240-246.

Solorzano, L.  1969.  "Determination of Ammonia in Natural Waters by
  the Phenolhypochlorite Method."  Limnol. and Oceanog. 14_(5) :799-801.

Sugam, R. and G.R. Helz.  1976.  "Speciation of Chlorine Produced Oxi-
  dants in Marine Waters."  Presented at Workshop on the Fate and
  Effects of Chlorine on the Marine and Estuarine Environment, Univer-
  sity of Maryland, Solomons, Maryland.  March 16-18.

White, G.C.  1972.  Handbook of Chlorination.  Van Nostrand Reinhold
  Comp., New York. 744 pp.

Zadorojny, C., S. Saxton, and R. Finger.  1973.  "Spectrophotometric
  Determination of Ammonia."  Water Poll. Control  Fed.   45:905-912.
                                224

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DISCUSSION

     Dr. Riley;  What kind of percent error do you get when you use
the ampiometric titration method?  What is the percent deviation for
your performance of triplicate analyses of the same sample?

     Mr. Hergott;  We could not perform triplicate samples during the
power plant monitoring because the level that comes through at the
condenser is always varying.

     Dr. Riley;  Are you really positive that you have a two-fold
difference there in your comparison?

     Dr. Hergott;  We saw the two-fold difference with the DPD method
and the ampiometric method.

     Dr. Baumgartner;  Did you have any trouble getting information
on chlorine use from the power plant officials?

     Mr. Hergott;  They were generally helpful.

     From the floor;  Has the state of California shown any interest
in changing their standards?

     Mir. Hergott:  I have not heard anything about that yet.  They
are considering an appeal by PG&E to increase the 0.0 level.  PG&E
performed substantial work and presented data trying to change this;
in  fact, we obtained the data on receiving water studies from
this work.
                                 225

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     THE NBS PROGRAM FOR STANDARDS FOR TRACE ORGANIC ANALYSIS
                    IN THE MARINE ENVIRONMENT
          H.S. Hertz, S.N. Chesler, W.E. May, S.A. Wise, L.R. Hilpert,
                 J. Brown, A. Fatiadi, and F. Guenther*
     The National Bureau of Standards  (NBS) is currently conducting a
research program to develop standards  for trace organic analysis in the
marine environment.  In this paper, we summarize the results of these
analyses for petroleum-in-sediment and petroleum-in-biota reference ma-
terials, marker compounds, and concentration techniques for polar or-
ganic materials.
PETROLEUM-IN-SEDIMENT REFERENCE
MATERIALS
     As previously reported to Environmental Protection Agency  (EPA),
a limited interlaboratory sample split was- conducted to determine the
suitability of sediment  (^1 ppm hydrocarbon burden) from the Katalla
River, Alaska, as a petroleum-in-sediment intercalibration material.
The results of this sample split encouraged the initiation of a more
extensive intercalibration exercise.
              

     An NBS sampling party collected ^100 kg of intertidal sediment
from the Katalla River area and from a relatively pristine Alaskan re-
gion, Hinchinbrook Island.  The samples from each site were homogenized
by mixing in a clean cement mixer for 3 hr, and samples of ^300 g each
were removed from the rotating mixer and placed in bottles.   (Except
for this brief period of homogenization, the samples were frozen from
the time of collection.)  Two bottles of sediment from each site were
sent to 12 laboratories for hydrocarbon analysis by the laboratories'
respective methodologies.  The participating laboratories include four
National Oceanographic and Atmospheric Administration  (NOAA) contract
laboratories  (NOAA is partially funding the study), two EPA labora-
tories, two Bureau of Land Management contract laboratories, and aca-
demic, industrial, and private laboratories.

     Samples were analyzed at NBS by headspace-sarapling gas chromatog-
raphy  (GC), gas chromatography and mass spectroscopy  (GC-MS), and
*    National Bureau of Standards, Analytical Chemistry Division,
Washington, D.C. 20234.

                                227

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 liquid chromatography  (LC) techniques.  Since all the other  laboratories
 use some type of  solvent extraction to remove the hydrocarbons  from  the
 sediment matrix,  samples were analyzed by an extraction method  for com-
 parison.  The comparison of headspace-sampled and solvent-extraction
 values will determine  if a bias exists for particular types  of  compounds
 between the two methods.

     For this intercalibration exercise, the following data  were com-
 pared:

 1.   Total hydrocarbons in GC elution range  (approximately C10  - C30)

 2.   Total extractable hydrocarbons

 3.   Pristane/phytane ratio and the amount of each present

 4.   Percent water

 5.   Identities and amounts of the three most abundant aliphatic and
     three most abundant aromatic hydrocarbons

 6.   Total polynuclear aromatic hydrocarbon  (PAH) concentration (four
     rings and larger)

 7.   Identity and amount of the most abundant PAH (four rings or lar-
     ger) .

     The comparison of these data from the participants should provide
 a measure of the  state of the art in trace hydrocarbon analysis.

     Since some of the participating laboratories have not yet re-
 sponded, the results of this intercalibration exercise will  be included
 in the next report to EPA.
PETROLEUM-IN-BIOTA REFERENCE
MATERIALS	

     The ultimate goal of this project is to release a biological trace
organic standard reference material.  However, immediate efforts have
been directed toward the preparation and release of homogeneous mate-
rials contaminated with low levels of petroleum.

     In conjunction with this effort, samples of Elizabeth River
("dirty")  and Middle Marsh ("clean") oysters were obtained from  Dr.
Robert Huggett of Virginia Institute of Marine Sciences and Dr. F.
Cross of National Marine Fisheries Services, respectively.  Both sets
of oysters were analyzed by a headspace-analysis GC procedure  (see NBS
Tech. Note 889) and by a modified procedure involving a high pressure
                                228

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liquid chromatography  (HPLC) clean-up of the TENAX column prior  to GC
analysis.  Results of these analyses are shown in Exhibit 1.
            EXHIBIT 1:  GAS CHROMATOGRAMS OF ELIZABETH
              RIVER (A) AND MIDDLE MARSH  (B) OYSTERS
     After headspace analysis, the residues left in each flask were
extracted with n-pentane.  The extracts were reduced in volume and ana-
lyzed for polynuclear aromatic hydrocarbon  (PAH) content by HPLC, using
ultravoilet and fluorescence detection.  Exhibits 2 and 3 show LC elu-
tion profiles of the two oyster samples.  The total PAH content, calcu-
lated in terms of 254 nm chrysene equivalents (i.e., assuming an absorp-
tivity equivalent to that of chrysene at 254 nm), was 0.5 mg/kg for the
Elizabeth River oysters and 0.05 mg/kg for the Middle Marsh oysters.
Two of the individual PAHs (chrysene and 3,4 benzopyrene) in the
Elizabeth River oysters were identified on the basis of retention vol-
ume and fluorescence emission spectra (see Exhibit 2).
                                229

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          A
      UV 254 nm
   n
 UU
        Fluorescence
          ex-290 nm
          em ' 400 nm
JX/"iV^
                                                            VX^
EXHIBIT  2:   HPLC  ELUTION  PROFILES  OF ELIZABETH RIVER OYSTERS
                                              Fluorescence
                                              ex-270 nm
                                              em-3SO nm
EXHIBIT 3:   HPLC ELUTION PROFILES OF MIDDLE MARSH  OYSTERS
                            230

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MARKER COMPOUNDS

     In this section, we describe marker compounds used as internal
standards and used for the qualitative identification of petroleum.

     Marker Compounds Used As Internal
     Standards	

     The height and area of chromatographic peaks are affected riot only
by sample size, but also by factors that influence the sensitivity of
response of the detector, such as the flame ionization detector and the
flame geometry, on column interactions or the molecular structure of
the solute.  If quantitative results are to be obtained, these varia-
tions must be carefully controlled.  The effect of a variation must be
eliminated by an appropriate compensation technique, such as the inter-
nal standard method.

     Most of these variables can be nullified by referring peak height
or peak area ratios to an internal standard.  Use of an internal stan-
dard compensates for minor variations in column operation or detector
response because both the internal standard and the sample are in-
fluenced to essentially the same extent.  The concentration of any com-
ponent can be calculated, using the following equation:

                   K. C. R
                 _  is is c
               c ~ R. K
                    is c

where:

      K.  and K  = detector response factors
       is      c
      R.  and R  = measured detector signals of the
       is      c
                   external standard and the sample
                   component, respectively

              C.  = known concentration of the inter-
                   nal standard.

In many cases where high accuracy (<5% error)  is not required or the
measurement of response factors is impossible or impractical, these
response factors are arbitrarily assigned a value of unity.

     An ideal internal standard will meet the following criteria:

1.   It should yield a completely resolved chromatographic elution
     profile.

2.   it should have an elution time nearly equal to that of the com-
     pound being measured.
                                231

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3.   The ratio of its peak height or area to that of the compound being
     measured should be close to unity.

4.   It should not be present in the original sample.

5.   Chemically, it should act similarly to the compound being measured.

6.   It should have a response factor equal to that of the compound be-
     ing measured.

7.   It should be in equilibrium with the matrix that contains the com-
     pound being measured.

8.   It should be added to the sample at the time of collection to com-
     pensate for losses of sample components.

     The advantages of the internal standard technique are that sample
size need not be known, and reasonable accuracy may be achieved without
the exact reproduction of operating conditions from run to run.*

     The requirements of an ideal internal standard are never realized
in the analyses of real samples, with the possible exception of isotope
dilution experiments.  Therefore, certain compromises must be made when
choosing an internal standard compound.  The simultaneous use of more
than one internal standard is a viable method that we have used for
trace-level petroleum analyses.

     Internal standards can be used to relate separate analyses on an
absolute basis.  For example, analyses of an environmental sample from
an oil spill and of unweathered spill oil can be corrected for differ-
ences in extraction efficiencies through use of the same internal stan-
dard.  Representative chromatograms can thus be directly compared, and
the weight % of oil in the environmental sample can be calculated, using
the following equation:
     I,  h    We   he .
         n          is
     E  he   W   h.
          n        is
    n=a
                           100 = wt % oil in sample
where hn = peak height of normal aliphatic hydrocarbon of carbon number
           n in sample chromatogram
*    Parts of this discussion of the interanl standard method were
adapted from Dal Nogare, S. and R.S. Juvet, Jr., 1962.  Gas-Liquid
Chromatography:   Theory and Practice, Interscience, New York.
                                232

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           he  = corresponding peak height from unweathered spill-oil
                 sample chromatogram

           h.  = peak height of internal standard in sample chromato-
                 gram

           he. = corresponding internal standard peak height in unweath-
                 ered spill-oil sample chromatogram

           W   = weight of sample analyzed

           We  = weight of unweathered spill-oil sample used as external
                 standard.

     Marker Compounds for Qualitative
     Identification of Petroleum

     Seven samples were chromatographed on a 3.05 m (10 ft) x 2 mm glass
GC column packed with 1% SE-30 coated on chromosorb WHP.  The column
effluent was split 1:1 into a flame ionization detector (FID)  and a
flame photometric detector (FPD).  The FID is a universal organic detec-
tor; the FPD will respond only to compounds containing sulfur (or phos-
phorus, depending upon detector setting),  Exhibits 4-10 are the dual
chromatograms obtained  (FID and sulfur FPD).  The similarity in sulfur
chromatograms between the Kuwait crude oil and the tanker spill oil is
particularly notable.  The spill oil was identified as a light Arabian
crude oil, suggesting that sulfur fingerprints might be an easy method
of identifying- sources of oil pollution.
CONCENTRATION TECHNIQUES FOR POLAR
ORGANIC COMPOUNDS	

     Initial liquid chromatographic separations of some nitrogen hetero-
cycles and some phenolic compounds have been achieved, and a program in
organic synthesis to prepare some of the larger hydroxylated aromatic
compounds is underway.  These compounds must be characterized before
developing separations procedures for environmental samples.

     Liquid Chromatographic Analyses

     Liquid Chromatographic methods have been developed for the analysis
of two groups of polar compounds:  phenols  (including a naphthol) and
N-heterocyclic compounds.  These two groups are of current environmental
interest because they are present in energy sources such as petroleum,
synthetic oil, shale oil, and coal.  Liquid Chromatographic separations
for both classes of compounds have been achieved, using both normal-
phase and reversed-phase liquid chromatography.
                                233

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EXHIBIT 4:  GAS CHROMATOGRAMS OF SOUTHERN LOUISIANA CRUDE OIL
   (Upper trace, flame ionization detection;  lower  trace,
  flame photometric detection)
EXHIBIT 5:  GAS CHROMATOGRAMS OF KUWAIT CRUDE OIL
   (Upper trace, flame ionization detection; lower
  trace, flame photometric detection)
EXHIBIT 6:  GAS CHROMATOGRAMS OF NO. 2 FUEL OIL
  (Upper trace, flame ionization detection;
  lower trace, flame photometric detection)
                           234

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EXHIBIT 7:  GAS CHROMATOGRAMS OF BUNKER C FUEL OIL
   (Upper trace, flame ionization detection;  lower
  trace, flame photometric detection)
EXHIBIT 8:  GAS CHROMATOGRAMS OF KUWAIT CRUDE OIL
   (Upper trace, flame ionization detection;  lower
   trace, flame photometric detection)
EXHIBIT '9:  GAS CHROMATOGRAMS OF NO.  2 FUEL OIL
   (Upper trace, flame ionization detection;
  lower trace, flame photometric detection)
                              235

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      EXHIBIT 10:  GAS CHROMATOGRAMS OF KATALLA RIVER SEDIMENT
         (Upper trace, flame ionization detection; lower trace,
        flame photometric detection)
     The separation of a synthetic mixture of phenols using the
reversed-phase mode (y Bondapak C18) and the normal phase mode (y Bonda-
pak NH2) is illustrated in Exhibit 11.

     Reversed-phase LC was also suitable for the analysis of a synthetic
mixture of alkyl-substituted N-heterocyclic compounds,  as shown in
Exhibit 12.  These compounds are representative of the  types of
N-heterocyclics commonly found in the same energy sources.  This same
mixture can also be separated on a p Bondapak NH2 column, using a non-
polar mobile phase as illustrated in Exhibit 12.

     Synthesis of Hydroxylated Aromatics

     The organic synthesis program started with the preparation of mono-
and dihydroxyanthracenes.  A review of the literature revealed several
ways of synthesizing mono- and dihydroxy derivatives of polycyclic aro-
matic hydrocarbons.  The first route involved careful sulfonation
(fuming sulfuric acid) of a hydrocarbon.  However, for the preparation
of 1-anthrol, anthraquinone was sulfonated (in the presence of a mer-
cury II - salt catalyst) to give an intermediate 5,10-anthraquinone-l-
sulfonic acid  (isolated as the K-salt).   Subsequent reductive elimina-
tion (treatment with warm ammonium hydroxide and zinc dust) and hydroly-
sis (potassium hydroxide fusion) converted the intermediate into a some-
what air-sensitive 1-anthrol.  A direct sulfonation of anthracene at
100-110C produced anthracene-2-sulfonic acid, which was converted into
relatively stable 2-anthrol.  When the direct sulfonation was carried
out at 150-170C, the main product was  anthracene-l,5-disulfonic acid;
this was transformed by hydrolysis  (i.e., fusion) into anthracene-1,5-
diol.   All compounds  (except 1-anthrol)  were purified by column chroma-
tography [Florisil, 1:1  (v/v) ethyl acetate-benzene or silica gel, 1:1
(v/v)  glacial acetic acid-benzene].
                                 236

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EXHIBIT 11:  SEPARATION OF  SYNTHETIC  MIXTURE OF PHENOLS USING
  y BONDAPAK CXB  (A) WITH 40%/60%  CH^CN/I^O AS THE MOBILE
  PHASE AND la BONDAPAK NH2  (B) AS  85%/15% CH2CL2/HEXANE AS
  THE MOBILE PHASE  ['(a) phenol,  (b) g-cresol,  (c)  o-cresol,
   (d) 2-naphthol,  (3) 2,4,6-trimethylphenol]
                             237

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                                       u
EXHIBIT 12:   (A) SEPARATION OF N-HETEROCYCLIC MIXTURE ON
  y BONDAPAK C18 WITH LINEAR GRADIENT OF 25-100% CH3CN
  IN H20 IN 40 KIN  [(a) 2,5-dimethylpyrole,  (b) 4-ethyl-
  pyridine, (c) lepidine, (d) 6-methyl 1,2,3,4-tetra-
  hydroquinoline,  (e) 2,3-dimethyldibenzocarbazole];
  (B) BLANK GRADIENT FROM 25-100% CH3CN IN H2O IN  40 MIN;
  (C) SEPARATION OF N-HETEROCYCLIC MIXTURE ON y BONDAPAK
  NH2 (with linear gradient of 0-35% CH2CL2  in pentane  in
  40 min at 2 ml/min)  [(a) through  (h) same  as in  (A)].
                           238

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     A second procedure for preparing hydroxylated  aromatics  involved
osmium tetraoxide hydroxylation of a hydrocarbon  (to  give  a cis-diol)
and careful acid-catalyzed elimination to yield a mono-hydroxy product.
Using this procedure, pyrene was hydroxylated  and cis-4,5-pyrenediol
was hydrolyzed to mono-hydroxy pyrene.  This procedure was also  applied
in preparation of some unique polyhydroxylated aromatics  (e.g.,  tetra-
hydroxyanthracene).

     The third procedure for preparing hydroxylated aromatics involved
oxidation  (chromic or periodic acids) of polycyclic,  aromatic hydrocar-
bons to quinones, which were converted either  to trans-diols  (via  sodium
borohydride reduction) or aromatic hydroxy-compounds  (via  sodium dithi-
onate treatment).  Using this procedure, phenanthrene, pyrene, triphen-
ylene, and chrysene have been oxidized to quinones.   These intermediates
must now be converted to the hydroxy compounds.

     The purity of all intermediates and new compounds prepared  was
verified by TLC, infrared, and mass spectrometry.   Future  work will
include improvement in the procedures for sulfonation and  fusion,  im-
proved osmium tetraoxide hydroxylation  (including decomposition  of the
osmate ester), and acid hydrolysis in the aromatic  cis-  and trans-diols.
DISCUSSION

     From the floor:  Did you say what the results of your analyses
were?

     Dr. Hertz:  We have the results from 7 of the 13 laboratories.
The results varied according to the parameter reported.  For example,
while everybody agreed on water content within a few percent, the total
extractables varied over a range of roughly 50 (which we considered
reasonably good) at the low ppm or ppb levels.

     From the floor;  When do you expect to come out with your first
standard, and what will that be?

     Dr. Hertz;  I would assume that the first standard might be some-
thing on the order of these water columns that I was discussing.  Maybe
in 2 years we could complete the stability test and various other stud-
ies for such a sample.  We hope that this would be a certified standard
reference material:  one at which we had actually certified the levels
with a certain level of confidence.

     Dr. Riley;  Have you decided what type of criteria you would place
on the solubility of aromatic compounds in, say,  distilled water?  For
example, have you done any kind of experiments in which you have actu-
ally filtered the water to which you have added the aromatic compounds
to see if there are any changes in the concentrations?
                                239

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     Dr. Hertz:  No.  Using these so-called generator columns, we have
pumped at various rates through the columns and have found that the
concentration is essentially independent of the pumping rate.  We have
also subsampled as little as 100 mfc and have found the concentration to
be the same within this amount and the batch.  I have not examined
whether this is dispersed or solubilized material.  We assume that it
is dissolved.

     Dr. Riley:  When you do analyze these samples, do you just break
the ampule open and then take a sample out?

     Dr. Hertz:  That is what we did in the initial case with the ampule
material.  Generally with the generator columns, we have been monitoring
the effluent from the generator column directly in a UB detector or an
LC system.

     Dr. Dieter:  I have been collecting data from petroleum hydrocar-
bons in different invertebrates.  Have you published data on the con-
centration of these various substances in shellfish?

     Dr. Hertz:  No.  We do have a manuscript describing the analytical
technique that we have been using for the hydrocarbons in biota; in
that, we give some data for various shellfish samples.

     From the jfloor:  How are you planning on homogenizing the tissues?
Are you planning on freeze-drying them or wet freezing, or what?

     Dr. Hertz;  We will first study some middle samples, under the
joint sponsorship of the EPA program and the NOAA/BLM program.  At this
point, we intend to thaw the samples, homogenize them in an ultrasonic
probe in the presence of KOH, refreeze them, and release a digest,
instead of freeze drying.

     Dr.Richards:  I would like to share with you some of the experi-
ences that we have had at the Gulf Breeze Laboratory which sound very
similar to the types of high-pressure liquid chromatography system that
you have been using.  We have found that it is very helpful to use
appropriate elution gradients when working with the microbondapack C-18
column.  I assume that you did use gradients.

     Dr. Hertz;  C-2 nitrile water gradient.

     Dr. Richards:  Exactly.  By varying the type of gradient and the
concentrations, we have been able to separate krysene and other things
from very complex mixtures.  We have also been using dimethylsulfoxide.
If you do use dimethylsulfoxide gradients with this particular column
(the C-18 microbondapack reverse phase), you must maintain a high,
uniform temperature in the column to keep the flow rate appropriate.
In that way, you can perform some separations that you cannot get with
the aceto nitrile.
                               240

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     Dr.  Hertz:   We have not worked with dimethylsulfoxide.   However,
we can put a sample on a microbondapack NH2  column and  do normal  elu-
tion with pentane.  By doing this, we can get a separation by ring  size
off the microbondapack NH2; "we can separate  aliphatics  from  the one
ring and the alkalated one-ring compounds, which come off in one  piece,
then the two-ring, the three-ring, and four-ring, and so on.   As  a  mat-
ter of fact, within the four-ring, we begin  to get some separation  of
the different four-ring polymeric hydrocarbons off the  microbondapack
NH2; but again,  always eluting in the same peak, to the alkalated and
the non-alkalated parent of PNA.  Those fractions can be put on a micro-
bondapack CAT reverse-phase column to obtain beautiful  profiles sepa-
rating out.  You can actually obtain flourescent submissions  for  iden-
tification.  We can also take those fractions and put them on GC-MS.

     Dr. Richards:  Right.  In field materials, we found a peak that is
very similar to crysene.  I just want to point out the  hazard of  find-
ing peaks in the field that appear to be very similar to those others.
In particular, we have been working with preparation of samples from
lugworms, shellfish, fish, and algae.

     You also mentioned the fluorescence spectrum work.  We  have  had
orders of magnitude of increased sensitivity using fluorescence.  In
addition, we have been working with Dr. Bens of the Coast Guard,  who
has a rapid scanning device that can be used to continually  measure
the fluorescent spectrum as it is eluted from the high-pressure liquid
chromatogram.

     Dr. Hertz:  The identifications made on the basis  of fluorescent
emission spectra were all confirmed, at least to the extent  of molec-
ular weight by the GC-MS.

     From the floor:  You mentioned that you are 2 years away from  a
certified standard?  What about an interim standard?  Are you inter-
ested in working on the development of a standard that  could be issued
in a lot less time but without the quality of a certified standard?

     Dr. Hertz:  We are interested in something on the  equivalent of
research material.  This is essentially what we are looking  at in terms
of the sediment and the biota.  I do not know that we really  want to
get involved in water because we are very concerned about adsorption on
containers, and so on.  I do not know that we could even say  something
was homogeneous and hope that people using different methods  could  come
up with the same answer on something that was delivered in a glass
ampule.

    'Dr. Lefcort:  I believe the question was, when could we  expect to
have research materials, interim standards?

     Dr. Hertz;   In terms of sediment, there are limited quantities of
materials around now, through the auspices,  mainly of the OCS work.
                                241

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We have supplied samples to several EPA laboratories as well as several
university laboratories.  We have not given very large-scale sediment
material much consideration yet, nor are we being asked for
intercalibration.
                                242

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       THE NBS PROGRAM FOR STANDARDS FOR TRACE INORGANIC ANALYSIS
                        IN THE MARINE ENVIRONMENT

                    I.L. Barnes, H.L. Rook, and T.J. Murphy*
     In the past several years, a particular emphasis has been placed
on new materials to be used in the assessment of environmental problems
associated with the production of energy from a wide variety of possi-
ble sources.  The Analytical Chemistry Division of the National Bureau
of Standards (NBS) has been conducting a research program concerning
the production of new standard reference materials and the determination
of new materials that can be used as standards in the analysis of inor-
ganic elements.

     The paper focuses on the use of raw materials, products, and resi-
dues of coal conversion and oil shale processes for possible standard
reference materials; the development of standards for monitoring water
quality associated with energy production; and the development of a
resin disk preconcentration method and an improved analytical capabil-
ity based on neutron activation.
ANALYSIS OF RAW MATERIALS, PRODUCTS,
AND RESIDUES OF COAL CONVERSION
AND OIL SHALE PROCESSES	

     In this section, we describe ongoing research on standard refer-
ence materials for coal and coal fly ash, oil shale, and the evaluation
of source materials.

     Coal and Coal Fly Ash

     Coal-SRM 1632, Coal, which is certified for trace element concen-
trations, has been sold out.  Two coals of different trace element con-
centrations have been obtained to replace this standard.  One is a
western sub-bituminous coal, and the other is an eastern bituminous coal.
In general, the eastern coal is about an order of magnitude higher in
*    Analytical Chemistry Division, National Bureau of Standards,
Washington, D.C. 20234.
                               243

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trace element concentrations than the western coal.  Each of these
materials has been ground and sieved.  The -65 sieve fraction was taken
for each standard reference material.  After blending, nine samples
were taken from each coal, and the homogeneity of each proposed stan-
dard reference material was determined by using neutron activation
analysis to examine eight elements.  The results of these determina-
tions indicate that the western coal is homogeneous within the preci-
sion of measurement (2%).  The homogeneity study of the eastern coal
is currently in process.  Upon completion of this work, each proposed
standard reference material will be packaged and samples taken for
trace element determinations.

     A study of the moisture content and drying of these coals has
been completed.  Although the western coal was found to have an
extremely high moisture content (18%), both coals can be dried by
vacuum drying for 24 hr at room temperature or by freeze-drying for the
same time.  All analyses for certification will be on a dry weight
basis.

     SRM 1633, Coal Fly Ash, is also out of stock and in need of re-
placement.  A replacement material has been located and ordered.  Prep-
aration and certification of this material are of high priority and"
will begin upon receipt of the material.

     Oil Shale Standard

     Two candidate oil shale materials have been obtained.  One is a
25 gal/t shale, and the other is a 35 gal/t shale.  They have each
been ground to two mesh sizes:  -65 mesh and -200 mesh.  The -65 mesh
materials are intended as reference materials for the Fisher assay.
The -200 mesh materials are intended to be trace element standard
reference materials.  A drying study on this material has been com-
pleted, and the samples surveyed for trace element profiles; however,
further work has been delayed because of the recent de-emphasis of
shale oil production.

     Source Material for Candidate
     Standards for Coals and Oil Shale

     Two representative coal and two representative oil shale mate-
rials were analyzed for trace element profile using a newly developed
multielement procedure.  The data were used to determine similarity of
material to the expired coal SRM 1632 and to ensure that the trace
levels between materials were not redundant.   The results of this sur-
vey are given in Exhibits 1 and 2.
                               244

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EXHIBIT 1:  ELEMENTAL CONCENTRATIONS  (yg/g UNLESS INDICATED)
            AND ESTIMATED UNCERTAINTIES ATTAINABLE
SMI 1632
Penn Coal
Ma (*)
K (%)
Rb
Cs
Sr
Ba
P,r
Sc
Cr
i:c r%)
Co
As
So
Sb
La
Co
Sm
Cu
Yb
Lu
Hf
Ta
Th
0
0
30
2
90
1 4 0
56
5
35
1
6
9
2
0
16
30
2
0
0
0
1
0
4
.089 
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   EXHIBIT 2:   ELEMENTAL CONCENTRATIONS (yg/g UNLESS INDICATED)
              AND ESTIMATED UNCERTAINTIES  ATTAINABLE
SRM 1632
Oil Shale
Na (.)
K (4)
Rb
Cs .
Ba
Sc
Cr
Tc (*)
Co
As
Sc
Sb
La
Co
Sra
Ku
Yb
Lu
Ilf
Ta
Th
1.
1.
63
3.
540
4.
32
1.
9.
64
3.
2.
20
34
2 _
0.
0.
0.
1.
0.
4.
6 <55;
0 <5%

-------
elements (i.e., sodium, potassium, calcium, and magnesium), which are
in the low ppm range, act as typical matrix elements.

     The preparation of this material was based on the experience
gained during the preparation and certification of mercury in water
(SRMs 1641-1642) and on the results of stability and container studies
conducted jointly by the Analytical Spectrometry and Activation Analy-
sis Sections.  The results of this work indicated that a successful
standard reference material could be prepared at the levels indicated.
In addition, conventional polyethylene was indicated as a suitable low-
cost container material for the standard reference material, provided
the container was placed in a vapor barrier seal to prevent the loss of
water vapor.

     Two hundred and twenty liters of the standard reference material
were prepared in one batch and then bottled in specially cleaned con-
tainers.  The actual certification program was then initiated.

     The elements present have each been analyzed by two or more of the
following methods:  neutron activation analysis, isotope dilution mass
spectrometry, spark source isotope dilution mass spectrometry, atomic
absorption spectrometry, graphite furnace emission spectrometry, zeeman
atomic absorption spectrometry, inductively coupled plasma emission
spectrome'try, and polarography.  The results have been very gratifying
with only a few small exceptions.  These are being resolved and should
be completed in the near future.

     Finally, mercury was added to the original sample but without much
hope that it would be stable enough for certification.  It now appears,
however, that this may be possible and completed results are expected
when the standard reference material is available for sale by March
1977.

     Mercury in Water Standard

     The original mercury in water standard has been completely sold-
out.  Currently, a new set is being prepared.  Analytical work is
scheduled to start in March with certification completed by July 1977.

     Sea Water Standard

     The possibility of preparing a sea water standard reference mate-
rial is currently being explored.  This work is expected to include
explorations of such possibilities as issuance of a natural (sterilized)
sample, a freeze-dried material to be reconstituted by the user, and/or
a material representing the extraction of the essential trace elements.

     Suitable large storage containers have been obtained and cleaned.
These will be used to hold large (55 gal) working solutions for method
development.  Arrangements have been made to acquire, as needed, natu-
ral sea water of various salinities and various levels of natural
                                247

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contamination.  The first material, a clean estuarine (half-sea) water
(salinity values of 17-20), will be obtained in the near future.  Work
to date has concentrated on the development of analytical procedures
for the heavy metals in sea water.

     Oyster Tissue Standard

     Since oysters are known to concentrate the trace metals present in
their environment, they have been proposed for a standard reference
material.  Sufficient oysters for a standard reference material were
cleaned, freeze-dried, ground, and sieved.  Samples were taken for
homogeneity testing by neutron activation analysis.  These analyses
showed that the material was inhomogeneous for a few elements.  However,
when the same oyster tissue was cryogenically ground (using liquid
nitrogen), sieved, and blended, the material was found to be homoge-
neous.

     Further work on this material was delayed because of the possibil-
ity that the International Atomic Energy Agency was going to distribute
a similar oyster tissue reference material.  However, they have aban-
doned this project since almost all of their material was used for
analyses.  Therefore, we plan to continue work on this proposed stan-
dard reference material.

     Ocean Sediment Standard

     An ocean sediment has been proposed as a standard reference mate-
rial for trace elements.  During the current fiscal year, we plan to
locate and acquire sufficient material for a standard reference mate-
rial.  The material will be ground, sieved, blended, and sterilized,
and its suitability as a standard reference material evaluated.

     Improved Irradiation Procedures

     Detection sensitivity for trace elements in water using neutron
activation analysis is dependent on the irradiation procedures for the
samples.  Past procedures for the irradiation of water samples in the
NBS research reactor were limited in sample volume to 1 mfc sealed in
quartz ampoules and to a maximum irradiation duration of 1 hr.  This
reactor safety limitation was due to the hydrogen gas pressure buildup
from the radiolysis of water.  Because of the limited quantity of water
that could be irradiated, the sensitivity for many elements was much
lower than normally expected for other matrix types.

     To overcome this problem, procedures were developed for the pre-
concentration of water samples by freeze-drying 10 to 100 m of the
sample and irradiating the residue.  Although this procedure did sub-
stantially improve the sensitivity for most elements, it was time
consuming and had potential problems because of the pre-irradiation
sample manipulations.
                               248

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     To overcome the problems with preconcentration of water  samples,
a new technique was developed, tested, and approved for the irradiation
of large volumes of water.  Polyethylene bags, made from  4 mil  thick-
ness, high-purity linear polyethylene, were  fabricated to match the
inside dimensions of the standard NBS irradiation  "rabbits."  The bags
were precleaned, filled with 15 m of water,  sealed,  and  irradiated
for periods of from 5 min to 4 hr in the high-flux pneumatic  irradia-
tion facility of the NBS reactor.

     The hydrogen pressure was found to reach pressure equilibrium
after 15 min.  The hydrogen diffusion through the  polyethylene  film
equaled the hydrogen production, and the bags maintained  physical in-
tegrity for more than 4 hr.

     This technique has been used for more than  6  months  and  has great-
ly facilitated the analytical procedure for  the  multitrace element
water standard.

     The Determination of Arsenic,
     Copper, and Antimony in Water

     Investigative work has been completed on the  accurate determina-
tion of the elements included in the multitrace  element water standard.
Using the new irradiation procedures, the elements silver (Ag), alu-
minum  (Al), cobalt  (Co),  chromium  (Cr), iron (Fe), molybdenum (Mo),
manganese  (Mn), strontium  (Sr), vanadium  (V), zinc (Zn),  selenium (Se),
and mercury  (Hg) could be determined nondestructively using various
irradiation, decay, and counting times.   Copper  (Cu), arsenic (As), and
antimony  (Sb) usually are not amenable to nondestructive  activation
analysis.  The most useful product radionuclides are  short-lived and
suffer  interference from  the dominant   Na  (sodium) activity  of the
sample.  Thus, these three elements require  chemical  separations from
the matrix activity.  Published separation procedures by  J. Costa et
al.  (1975),* for Cu, As,  and Sb are capable  of producing  accurate
results at the sub-ppm level of concentration.   However,  these  proce-
dures are highly dependent upon the pH of the extraction  system and are
not easily adaptable to multielement analyses.

     Among possible radiochemical procedures and reagents for group
separation of elements, inorganic ion exchanges  have  received increased
interest because of the high selectivity  that can  be  achieved.  Fur-
thermore, by separating elements in groups  instead of individually, and
utilizing the high resolution of semiconductor detectors, workers can
achieve simple low-cost and rapid analyses.
*    "Determination of  Purity  Levels  in  Ultrapure  Chemicals  by Activa-
tion Analysis," 1974-1975 Yearly  Report  of  Institute  of  Jozef Stefan,
Ljublijana, Yugoslavia.
                                 249

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     A chemical separation procedure was developed for the group  sepa-
ration of Cu, As, and Sb from water.  The procedure used two chromato-
graphic columns containing cuprous chloride and tin dioxide stacked in
series.  Following irradiation, samples were adjusted to a 6 molar con-
centration of HC104, and the resulting solutions were passed through
the columns at a flow rate of 2 m per minute.

     The separation scheme was tested on some botanical standard  refer-
ence materials, which had excellent certified values for Cu, As,  and
Sb.  These results are listed in Exhibit 3.

     EXHIBIT 3:  A COMPARISON OF NEUTRON ACTIVATION ANALYSIS
               RESULTS TO OTHER INDEPENDENT METHODS
Method
Ele-
ment
Spinach
Pine
Needles
Tomato
Leaves
Neutron Activa-
  tion Analysis      Cu    10.80  0.20    2.98  0.16   11.1 + 0.20

Atomic Absorption    Cu    12.70  0.40    3.20  0.40   11,5  0.20

Spectrofluorimetry   Cu    11.50           2.90          10.6

Neutron Activa-
  tion Analysis      As    0.120  0.01    0.20  0.02   0.23  0.03

Spectrofluorimetry   As    0.170  0.05    0.21  0.04   0.29  0.05

Neutron Activa-
  tion Analysis      Sb    0.043  0.004   0.22  0.01   0.12  0.03
     Following procedure verification, this separation was used for
certification analysis on the multitrace element standard reference
material.

     Certification Analyses for
     Trace Elements	

     The instrumental and separation procedures developed were used for
certification analyses on 13 of the elements in the new trace element
in the water standard reference material.  Because of their short half-
lives, Al, Mn, and V were determined individually.  Na, K, Mo, and Sr
were determined instrumentally as a group with moderate half-lives, and
Co, Cr, Zn, and Se were determined instrumentally as a second group
with long half-lives.  Cu, As, and Hg were determined, using radio-
chemical separations to isolate them from the matrix 2t*Na activity-


                              250

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     For certification, duplicate samples were  taken  from five  randomly
selected bottles and the replicate  samples  analyzed for  all  elements.
DEVELOPMENT OF A RESIN DISK
PEECONCENTRATION METHOD

     A number of workers have used a. commercially  available  chelating
resin (Chelex-100) to separate heavy  metals  from sea water by conven-
tional column chromatography.  This material seems  to have a number of
advantages over other types of resins in  that it chelates heavy metals
and does not strongly hold sodium, potassium,  calcium,  and magnesium
(the common sea water matrix elements), in most analytical procedures.
The column procedure, however, is inconvenient in  that  the elements
must then be stripped from the column and prepared  for  analysis.

     To eliminate these problems, we  have developed a method in which
a filter disk is manufactured and the chelating resin is incorporated
within the disk material  (not simply  layered on the surface).  These
disks, using standard filtering procedures,  permit  the  extraction of
the heavy metals from as much as 1   of water with  nearly 100% effi-
ciency.  Using radioactive forms of each  element, both  singly and in
groups, we have examined the efficiency of this technique.

     The advantages of the resin disk precedure are:

1.   Multielement analyses may be performed

2.   The sample may be easily preconcentrated up to a factor of 1000

3.   The samples exhibit long-term stability without special handling

4.   A wide variety of analytical methods may be used,  including sev-
     eral that can be used directly on the disk.

     For example, Exhibit 4 shows the x-ray  fluorescence spectrum of an
aqueous solution of 20 yg of a number of  elements  filtered through two
successive disks, and Exhibit 5 shows the same for  a sample  of 10 mg of
urban dust.*  These spectra were obtained directly  on the disk, the
base material of which offers no analytical  interferences.   Exhibit 6
illustrates the linearity of typical  analytical curves  over  a wide
range.
*    Note:  tungsten and  titanium  are  contaminants in the disk intro-
duced during manufacture.   These will  be eliminated in subsequent lots.
                                251

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                                           First Resin Disk
                                           Second Resin Disk
EXHIBIT  4:   AQUEOUS SOLUTION:   20 yg EACH METAL
  FILTERED  THROUGH SUCCESSIVE  RESIN DISKS
              Fe
                     TLA
                                       Background a Blank Disk
      EXHIBIT  5:   10 mg ST.  LOUIS DUST
        CHELATED  TO A RESIN  DISK
                      252

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                                                              /Zn K
        16-
        14
        12
        10
      X
      I 8

      O
      O
                                                              /PB L
                                                              /Pb L
                  20
                             40
                                        60
                                  Micrograms
80
                                                            100
              EXHIBIT  6:   NONINTERFERING MULTIELEMENT  ANALYSIS
                BY XRF OF  RESIN DESKS
     To date, we  have  used these disks for analysis by x-ray fluores-
cence, proton-induced  x-ray (directly),  and atomic absorption (by
removing the  elements  from the disks).  Activation analysis will be
used in the near  future.
DEVELOPMENT OF IMPROVED MULTIELEMENT
ANALYTICAL CAPABILITY	

     A neutron activation system has been  developed  and  evaluated  for
the accurate multielement analysis  of  diverse  materials.   This  type  of
approach optimizes the analytical information  obtained on  a series of
similar sample types while minimizing  the  manpower costs in the provi-
sion of this information.  Therefore,  this approach  has  potential  appli-
cability for more efficient  certification  analyses of environmental
standard reference materials.
                                253

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     Radiochemistry is not always required for determining many elements
in suitable matrices.  In the most favorable cases, it is only necessary
to irradiate samples and appropriate standards and to assay the result-
ing radioactivity with high-resolution semiconductor gamma detectors.
Savings in time and effort, especially for routine analyses, are con-
siderable, once the method has been developed to a desired level of
reliability.

     The equipment required for this type of analysis, in addition to
the NBS reactor, consists of Ge(Li)  detectors coupled with means for
automatic and accurate positioning of samples, multichannel pulse-height
analyzers with life-time and time-of-day clocks, and magnetic tape data
recorders.

     A crucial operation in any instrumental analysis is the reduction
of raw spectral data into final analytical results.  The analyst, in
cooperation with his surrogate, the computer, is required to judge the
qualitative and quantitative suitability of the data and select the
best of several possible means for analyzing the information at hand.
For our spectra, which usually contain 4,096 six-digit numbers from
which dozens of peak areas and positions must be extracted, a large
computer program is indispensable.  Spectral data are processed at the
NBS Computer Science Center by the computer code QLNl.

     This program performs extensive qualitative analysis of the data
to identify the peak characteristics of the isotopes sought.  The quan-
titative comparison of the appropriate peak areas in samples and stan-
dards to arrive at elemental concentration and error estimates is per-
formed for specified elements following qualitative analysis.   This
code is in a continual process of refinement, but the reproducibility
and accuracy are excellent for well-resolved peaks and clearly supe-
rior to hand calculations.  The analyst must still pay much attention
to the quality of the input data; however, this essential feature of
the instrumental analytical process is common to all methods of spec-
tral data analysis.

     A second feature of this system is the use of variable irradiation,
decay, and counting times to maximize the numbers of elements obtained
while minimizing the effects of interfering radionuclides.  Optimum
irradiation times, decay times, and counting times are selected on.the
basis of preliminary irradiation data and are applied to all other
analyses on similar sample types.

     The third and most essential feature of this system is that it has
been extensively evaluated for accuracy using existing NBS standard
reference materials as check samples.  These samples have been analyzed
in the multielement mode, and the results checked against the certified
concentrations of the individual element in the standard reference
material.  The system now yields consistently good data on materials
amenable to nondestructive neutron activation analysis.
                                254

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DISCUSSION

     Dr. Baumgartner;  Was one of  those early examples  in fresh water?

     Dr. Barnes:  That was in fresh water, yes.  However, we have since
sampled sea water at low parts per million.  We have used two different
methods, by the way:  activation analysis, which is the system of burn-
ing the material in a stream of oxygen; and an extraction technique.

     From the floor:  I am confused about the use of sodium, potassium,
magnesium, and calcium matrix metals in the fresh water samples.  Could
you explain their use for a standard?

     Dr. Barnes:  These materials  are  interferences.  If you are in a
laboratory which is normally analyzing water by biotonic absorption or
by activation analysis, you would  like a standard that  is a close  ,
approximation of the same matrix material you meet in your everyday
analysis; if we had left those out, we would have omitted a potential
interference.

     From the floor:  What if you  are  running an analysis on materials
that have a different matrix, for  example, water samples that have high
levels of hardness?

     Dr. Barnes:  If the material  is there in this level, the addition
of more to it does not create that much more of an interference.  In
other words, if you have a calcium interference, that is above this
level, it does not make much difference how far it is above it, within
reason.  As long as it is there, interference will result.

     Dr. Richards:  I am concerned about the massive amounts of these
salts that you are analyzing on the filter from the sea water samples.
You showed graphs with 20-100 mg of elements that naturally occur at
30 parts per trillion in that range.   Does it work at those levels?

     Dr. Barnes;  Yes.

     Dr._Richards;  What are your  flow rates?

     Dr. Barnes:  I have one here  that has 80 mg of those four elements.
I have the individual concentrations.

     Dr. Richards:  But none of them occur above 5 ppm  in natural sea
water?

     Dr. Barnes;  That is right.   We have also done the same thing with
natural sea water taken from the Virginia Institute of  Marine Science.
We are working right now on material that was gathered  for us about
7 or 8 miles out, which should be  much cleaner.
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     Dr. Richards:  What are the approximate flow rates that you run
through that filter?  We do Kelex extraction work, using columns, and
we typically run about 3-5 mills a minute.

     Dr. Barnes:  I am guessing that this is about 3 mills a minute,
but at what pH do you do that?

     Dr. Richards:  We do normal sea water.

     Dr. Barnes:  Yes, and that will not work.

     Dr. Richards:  I disagree with you heartily, sir.  It was published
in ELANO, 2 years ago.

     Dr. Barnes;  You do not get nearly the same extraction efficiency
as you do at a pH of about 5.2.

     Dr. Richards:  We do not modify our pH.  We have done tracer exper-
iments, to document that we do get complete recovery.  It actually does
work at those concentrations at 100 parts per trillion for cadmium.

     Dr. Barnes:  Yes.  You can also work with Kelox-100.  At a pH of
8.1 it acts as a healing ring.  You can actually make Kelox act like
a kelating agent; you can make it act like an ion exchange agent by
changing the pH; and you can, in fact, get a mixture of these two by
playing with the pH.  We have done a great deal of work with this.  Our
tracer experiments show that it varies by hpw much sodium, potassium,
calcium, and magnesium you are using, but you get a much better effi-
ciency, if you go to a pH of about 5.2.

     Dr. Richards:  Do you have any plans to make a standard reference
material of marine tissue?

     Dr. Barnes:  Yes.  As you may or may not be aware, we first tried
this with freeze-dried tuna fish, which did not work.  Despite our best
efforts, some of the bone was left in; the bone-seeking elements are not
homogeneously distributed.  The lead is reasonably well-distributed in
this particular material.  We have now prepared a sample of freeze-
dried oysters.  We initially had this material commercially prepared
for us and it seems extremely homogeneous.  We have found that, if we
regrind the material, pyrogenically, under liquid nitrogen, it makes an
extremely good standard.  We will start certification, probably within
a couple of months.

     Dr. Richards:  When you say "completely homogeneous," what sort of
standard deviation would you get for something well-behaved, such as
zinc or copper?

     Dr. Barnes:  At the Bureau of Standards, we try to analyze what
people in the field actually need and anticipate what levels of
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precision they are going to need in  the near  future.  We then try to
certify it at somewhere between three  to  ten  times better than that.
The standard must be somewhat better than everyday use.  For the well-
behaved materials, we are looking at something below 1 percent.  For
some of the others, we will settle for about  1 or 2 percent.
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STANDARDIZATION AND INTERCALIBRATION TECHNIQUES FOR MARINE MONITORING

                           Michael A. Basileo*
     The objective of this project is to develop standardization and
intercalibration techniques to assess the quality of marine environ-
mental data.  The tasks within the project are directed towards develop-
ing the capability to define the uncertainties or error boundaries
associated with data obtained from environmental measurement systems.
The project is composed of two tasks:

     1.     Standards development

     2.     Investigations.


STANDARDS DEVELOPMENT

     The various standards under development within this task can be
categorized by area of use, i.e./ laboratory, interlaboratory or trans-
fer, and field.  In each case, the development is directed towards pro-
viding a product needed to define or reduce the uncertainty levels in
a particular portion of the overall environmental measurement process.
Thus, while the individual subtasks may appear unrelated,  all support
the project goal of providing improved definition of environmental data
quality.

     Laboratory standards are, obviously, used in the laboratory as
the reference point for instrument testing and calibration.  In most
instances, these standards represent the laboratory's closest tie to
national defining standards and, as such, represent the base of the
overall measurement process, which culminates in field data.  We de-
scribe below the seven subtasks:  dissolved oxygen laboratory standard,
dynamic test apparatus, environmental test specifications, conductivity/
temperature/depth (CTD) transfer standard, pressure transfer standard,
CTD field standard, and a handbook of underwater optical instrumenta-
tion.
*    National Ocean Survey Test and Evaluation Laboratory, National
Oceanographic and Atmospheric Administration, Rockville, Maryland 20852.
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     Dissolved Oxygen Laboratory Standard

     Development of a laboratory dissolved oxygen standard is currently
in progress.  This standard will serve as a substitute for the Winkler
method, which is currently used as the reference in the test and cali-
bration of dissolved oxygen measurement systems.  The standard will
operate in a test bath over a range of -2 to 35 in temperature and
0 to 40 ppt in salinity.  The dissolved oxygen range will span 0 to 15
ppm, with a measurement uncertainty goal of 0.03 ppm.

     The Winkler method, which is a titration process, has demonstrated
shortcomings that have led to this effort.  Simplified, the measurement
process consists of sample withdrawal, sample preservation, and titra-
tion.  Each of these steps is extremely operator-sensitive; thus, it is
difficult to obtain repeatable data within a laboratory and extremely
difficult to obtain satisfactory agreement between different laborato-
ries performing the same measurements.

     The approach in developing the dissolved oxygen standard has been
to reduce the degree of operator intervention in the measurement pro-
cess.  The standard will directly monitor oxygen concentration in the
test bath by use of a submerged sensor.  The sensor is similar to the
polarographic membrane sensors presently utilized in field instruments.
The measurement principle, however, has been modified to eliminate the
errors caused by the membrane transport properties.

     The dissolved oxygen standard is being developed under a contract
with the University of South Florida.  The first phase of the develop-
ment will result in a prototype sensor, which will undergo testing at
the test and evaluation laboratory in March 1977.  Satisfactory perfor-
mance of the prototype sensor will lead to completion of the final sys-
tem and acceptance testing, which is scheduled for December 1977.  The
subsequent documentation of a proven alternative to the Winkler titra-
tion method is expected to encourage adoption of the standard by other
laboratories and ultimately to improve the traceability of dissolved
oxygen data.

     Dynamic Test Apparatus

     A dynamic test apparatus for laboratory testing of flow sensors is
under development.  The apparatus will generate controlled dynamics
superimposed on steady flows of up to 70 cm/sec to establish flow
sensor capabilities in the nonsteady flow environment.  Present methods
of testing water current sensors by determining their output over a
range of steady-state speeds yield little information on the sensor
performance under dynamic conditions.  Dynamic conditions, however,
represent real-world conditions in the marine environment when the
sensor is subjected to motions induced by moorings, turbulence, and
wave-generated water motions.  Some initial efforts in the dynamic
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testing area have indicated that  these  errors can range from 20% to
well over 100%.  In attempting  to assess  the uncertainty levels associ-
ated with current measurements, it  is necessary  to have some knowledge
of the dynamic response characteristics of  the sensing system.

     The Naval Ship Research and  Development Center is designing and
fabricating the test apparatus, which will  be utilized on one of their
tow carriages.  The apparatus will  be capable of two degrees of freedom
and provide three modes of nearly sinusoidal dynamics:  horizontal,
vertical, and circular in a vertical plane.  The peak-to-peak amplitudes
can be varied from 0.4 to 1.2 m;  the period is adjustable over the range
of 2.5 to 12 sec.  The apparatus  can accommodate current sensors weigh-
ing up to 80 kg, to a maximum length of 1.8 m.   Inducing these motions
on the current sensor while the tow carriage is  underway provides a
simulation of steady current with superimposed fluctuations.  To date,
the vertical circular mode has  been completed and tested.  Hardware
fabrication for the remaining two modes has been completed, and accept-
ance testing is to begin in March 1977.   A  final report documenting the
overall dynamics simulation system  will be  completed in August.  The
apparatus can then be utilized  in defining  the errors in current
measurement caused by dynamic effects.

     Environmental Test Specifications

     Environmental test specifications  will be developed to provide a
higher operational reliability  for  marine instrumentation.  Presently,
no standardized set of environmental test specifications exists that
is directed toward marine instrumentation.  Consequently, instrument
designers do not have sufficient  information about the environment in
which their product is expected to  operate.  The usual result is under-
design rather than overdesign,  a  fact that  has been documented by past
testimonials on poor instrument performance.  The solution to this
problem lies in telling the instrument  supplier  what environmental con-
ditions his product will encounter  and  having him demonstrate in a test
laboratory that his product can survive these conditions with a high
degree of certainty.

     The environmental test specifications  to be developed will provide
the information on conditions encountered in typical marine-monitoring
operations, which include shipping, shipboard storage and handling,
temperature, humidity, shock, vibration,  and salt spray.  The specifi-
cations will be focused on the  environment  encountered in operations 
from typical marine research vessels ranging in  size from the launch up
to about 100 m in length.  Future efforts will include buoys and other
field environments in which instrumentation is used.

     Work statement development will commence in the second half of
fiscal year 1977 with a goal of awarding  a  contract by the end of the
year.  Completion of the contract is expected within 1 year after award.
The resultant environmental test  specifications  would then be available
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for inclusion as part of an instrument procurement package and for use
in performing the subsequent acceptance testing,

     Conductivity/Temperature/Depth  (CTD) System

     Transfer standards are utilized to assess the comparability of
results obtained between different laboratories performing similar
calibrations.  Calibration of environmental sensors is an area in which
standardized procedures have not yet been fully developed.  As a con-
sequence, the calibration process tends to vary from one laboratory
to another.  Even when a common process is generally followed, varia-
tions in application tend to arise that have the potential of causing
errors in the final result.

     To define the uncertainty of a sensor calibration, which is a com-
ponent of the overall field data uncertainty, it is necessary to eval-
uate the total calibration process.  This can be accomplished by sev-
eral laboratories calibrating the same transfer standard  (i.e., a known,
highly stable sensor) and comparing results to expected values.  The
"expected" values result from calibration performed at the "reference"
laboratory.  This designation can be applied to a laboratory based on
its superior capability or can be applied arbitrarily to obtain a single
point of reference.  Results of the interlaboratory calibrations can be
used to evaluate the effect of variations in calibration process and as
a periodic check to verify that the laboratories in question remain in
some constant relation to one another.

     A conductivity/temperature/depth (CTD)  system was selected as one
of the transfer standard development areas.   These three parameters are
related to most environmental monitoring programs.  The measurement
goals of the transfer standard are 0.5 to 65 millisiemens/cm in con-
ductivity with an uncertainty of 5 microsiemens/cm;  -3 to 30C  with a
 mK uncertainty; and 0 to 1500 decibars in pressure with a  0.75
decibar uncertainty-

     An off-shelf CTD system was procured that showed the potential for
having the high stability required in a transfer standard.  Initial
measurements have confirmed this stability-   The system is presently
undergoing a laboratory evaluation over a 6-month time interval to
verify its measurement capabilities.  In the summer of 1977,  the  system
will be shipped to another laboratory for calibration.  It will then
be returned to the test and evaluation laboratory for final calibration
and data analysis.  The end result will be a report that defines  the
transfer standard capabilities and the procedures utilized for per-
forming intercalibration.   The measurement results obtained at the two
laboratories will be  presented as an example of a typical intercalibra-
tion experiment.
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     Pressure Transfer Standard

     The development of transfer standards and intercalibration tech-
niques for high-accuracy pressure  sensors is currently in progress.
These sensors, which are presently being employed in tide measuring
systems, have an accuracy approaching  that of the available standards.
Piston gauge pressure standards operated with careful technique are
marginally acceptable as calibration standards.  Slight variations in
the calibration process can  generate errors that are several times
larger than the allowable error of the calibration process.  The capa-
bility to perform interlaboratory  calibration will provide a means for
detecting these variations,  thus providing assurance that the calibra-
tion process remains within  narrow boundaries.

     Pressure sensors of the type  utilized in the high-accuracy tide
systems have b~en procured and will undergo a laboratory evaluation to
define their characteristics.  Pressure ranges of 0 to 10, 0 to 20, and
0 to 700 decibars were selected that correspond to the expected'"worst
case" measurement ranges in  the laboratory calibration process.  Upon
completion of the laboratory evaluations, the sensors will be calibrat-
ed at another laboratory and then  returned for final calibration and
data analysis.  The uncertainty goal for the interlaboratory comparison
is 0.01% of reading, which  corresponds to the expected uncertainty of
the laboratory standards utilized.  The final report, to be completed in
December 1977, will define  the  transfer standard capabilities and the
intercalibration techniques, and will  include the results of the inter-
calibration as an example.

     CTD Field Standard

     In the hierarchy of standards which shows the National Bureau of
Standards at the apex, field standards would usually be shown at the
lowest level.  The purpose of the  field standard is to provide some
measure of assurance that the monitoring instrumentation has not changed
characteristics during the course  of a field operation.  This capability
would not normally be adequate to  allow readjustment of a discovered
shift; it would, however, serve as a flag indicating that the quality
of subsequent data obtained  with the instrument is suspect.  For those
experimenters fortunate enough to  have spare monitoring instruments,
detection of a shift would  trigger instrument replacement.

     A field verification standard for CTD sensing systems will be
developed.  The use of the CTD  system  is common to many monitoring
activities; thus, the availability of  a field standard for this system
would provide improved definition  of quality for a significant volume
of marine environmental data.  The verification system to be developed
will provide an overall sensing system check.  It would simulate an
environmental parameter  (e.g.,  a particular temperature), which should
result in a certain system  output  within some defined tolerance level.
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     This development will begin in the second half of this fiscal
year; plans are to award the development contract by the end of the
fiscal year.  The final product, expected within 1 year after contract
award, would be the field verification system complete with the appro-
priate methodology for its use.

     Handbook of Underwater Optical Instrumentation

     The development of a users' guide for underwater optical instrumen-
tation has been put into the field standard category, but it is actually
an approach toward standardizing field methods.  Many types of commer-
cially available optical instruments are used in water quality measure-
ment programs.  Each type measures different optical properties, which
can sometimes be correlated with other water quality parameters.  In-
formation on the theory behind the various measurements is available,
but usually it is in a highly technical form which is not readily
usable by the scientist, engineer, or program manager unfamiliar with
optical theory.

     The purpose of this handbook will be to describe the basic theory
of operation of the various types of optical instruments, indicate the
kinds of quantitative and qualitative information that can be derived
from the provided measurements, and make recommendations on the best
approach for specific applications.

     A contract for the handbook was awarded to Scripps Visibility
Laboratory in October 1976.  Agreement has been reached on the final
handbook outline; the handbook itself should be completed by January
1978.  The completed text will be used to provide sufficient informa-
tion to scientists, engineers, and program managers so that they can
make an intelligent choice of optical instrumentation to meet the data
requirements of their monitoring program.  The text will also serve as
a basis for the standardization of optical property measurement methods.
INVESTIGATIONS

     In this task, we attempt to define the comparability of results
obtained from different approaches to the measurement of chemical
properties of marine waters.  The trend has been to develop direct
measurements (in-situ) systems to replace analytical laboratory methods
in water analysis.  The in-situ approach has several advantages; the
most significant is probably cost per measurement.  The accepted stan-
dards, however, are still defined in terms of analytical methods.  To
enjoy the benefits of in-situ monitoring without the loss of data
quality, we must first demonstrate the traceability of the newer methods
of the existing standards.  The results of such demonstration then serve
as a basis for defining the quality of data obtained from the in-situ
measurements.  This task includes the following subtasks:  methods for
salt water analysis, standards for in-situ chemical instrumentation,
water quality test kits, and specific ion electrode evaluation.

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     Methods for Salt Water Analysis

     The objective of this subtask  is  to  determine  the comparability
between different methods for  the chemical  analysis of salt water.  The
Environmental Protection Agency  (EPA)  publishes  standard methods for
chemical analysis, most of which are applicable  to  both fresh and saline
waters, in Methods for the Chemical Analyses  of  Water and Wastes  (1974).
The oceanographic community has  no  similar  standard text, but one of the
commonly accepted documents is A Manual of  Seawater Analyses by
J.D. Strickland and  T.B. Parsons (1968).

     In attempting to standardize a single  set of analytical methods for
saline water, it is  necessary  to determine  whether  conflict exists among
the results obtained by the various available methods and to assess
which of the methods provides  the most reliable  results in terms of
data quality.  The approach for  this work has been  to define 24 chemical
parameters from four categories: nutrients,  heavy  metals, low .molecular
weight hydrocarbons, and other common  analyses.  The parameters were
selected on  the basis of those most certain to meet the present and
future needs of the  National Oceanographic  and Atmospheric Administra-
tion  (NOAA)  and EPA  in  the area  of  sea water pollution monitoring.

     Proposals have  been requested  that require  the contractor to com-
pare the analytical  methodologies  for  the selected  parameters using both
open ocean and estuarine water.  Contract award  is  expected in March
1977; completion of  the  contract and analysis of results are scheduled
for September 1978.  The  final report  will  then  serve as a basis for the
recommendation  for  a standardized  set  of  salt water analytical methods
to be adopted by both  NOAA and EPA.

     Standards  for  In-Situ Chemical Instrumentation

     An  investigation  is  underway  to  assess whether there is measurement
traceability between in-situ instruments  and the corresponding analyt-
ical standards.  The increasing  availability of  in-situ instrumentation
for measurement of  chemical parameters has  resulted in significant
change  in operational  practices.  The  analytical methods being replaced
have been well  organized in terms  of documented  methods and standard-
ized procedures.   Instrumental techniques are not yet as well organized;
thus,  the  tie  or  traceability between  the two measurement approaches
remains  largely unproven.

      This  study will attempt to fill  the gap by determining the  chem-
ical parameters now being measured with in-situ instrumentation  and the
 standards  utilized to calibrate the instrumentation.  These standards
will be  compared to those established for the analytical methods; based
on the  degree of comparison,  the measurement uncertainties of  the
 in-situ  instrumentation will be estimated.   Recommendations for  changes
 in the  existing standards for instrumentation will  be provided where
 this would  upgrade the comparability of the two methods.
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     A contract was awarded for the study in June 1976.  A first draft
of the report is expected in July 1977; .the final report is due in
September.  The final report will provide guidance in developing the
quality control practices required to assure the traceability of in-situ
chemical measurement to accepted standards.

     Water Quality Test Kits

     An investigation has been initiated into the comparability between
results obtained from standard analytical methods recommended by EPA and
results obtained from the use of commercially available water quality
"test kits."  Inexpensive test kits for determining a variety of chem-
ical parameters are available and used by many for water quality
measurement.  The use of these kits has been questioned by EPA and
others, since little information is available on the quality of data
they provide and their applicability under various naturally occurring
background conditions.  The chemical parameters under consideration for
the comparison include ammonia, nitrate, dissolved oxygen, ortho-
phosphate, and total alkalinity.  Ten to fifteen specific kits will be
selected by mutual agreement with the contractor.  The tests will be
run on samples in several backgrounds:  fresh water, both clean and
polluted; and sea water, low and high salinity, both clean and polluted.
Contract award is expected by April 1977; completion of analysis and
report is'scheduled for March 1978.  The report will provide guidance
in the use and expected data uncertainties of water quality test kits
for chemical analyses in natural waters.

     Specific Ion Electrode Evaluation

     An investigation is underway to determine the applicability of ion-
selective electrodes to in-situ water quality measurements in natural
waters.  Many electrodes have been manufactured that are selective for
various chemical species and provide simple, low-cost methods of
analysis.  Several of these ion-selective electrodes have been used
to measure ions in natural waters.

     There are, however, several problems with electrode measurement
that have not been considered which jeopardize the quality of the data.
All electrodes are subject to some type of interference, particularly
in natural waters, and they do not respond directly to the total con-
centration in the sample but only to the free, "available" species.
These stability and calibration errors have not been studied, nor have
the response times, sensitivity to light, and flow variations been
determined.  There is much interest in using these electrodes as in-situ
monitoring sensors, but their characteristics in natural waters are
still undefined.  Before ion-selective electrodes can be effectively
used for in-situ applications, these problems and solutions must be
defined to ensure that the level of data uncertainty in these applica-
tions can be predicted.
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     To define the effects of these potential  sources of  error, a
series of evaluations of the ion probes  have been  initiated.  Testing
is already in progress on sodium, potassium, calcium, magnesium, and
chloride electrodes from several manufacturers.  Further  tests will
include fluoride and ammonia sensors  and two probes  for chloride and
fluoride, which are currently in use  on  water  quality monitoring sys-
tems.  The testing, analysis, and report are scheduled for completion
in October 1977.  The report will present,  for each  class of electrode,
results of the laboratory evaluations; thus, it will provide guidance
in the use and performance of selective  ion electrodes in in-situ
applications.
DISCUSSION

     Dr. McCammon;   Are  you going to recommend different manufacturers,
then, and put  a  stamp  of approval on different instruments?

     Mr. Basileo:   No.

     Dr. McCammon:   How  are you going to  separate out,  say, the kits
that will give reputable results and kits that will not?

     Mr. Basileo;   By  presenting the test results of the experiment.
We may end up  with  a particular kit  with  maybe three or four potential
manufacturers.   We  will  then present the  results of the test in terms
of how good  they were, and it would  be up to  the purchaser.  However,
there should be  enough data so we can tell that this one is clearly
better than  that one.

     Dr. McCammon:   I  was particularly interested to see what you had
been doing with  the CTD.   I know that the Navy has done an exhaustive
study on CTDs  to ascertain their reliability.

     Mr. Basileo;   When  you say "Navy," our group used  to be with Navy.
Our work was spurred on  by the fact  that  there are only one or two sys-
tems on the  market  that  have the potential stability that would be
usable for this  application.

     From the  floor:   Which instrument?

     Mr. Basileo:   We  selected the Neal Brown,  which is still being stud-
ied by the Navy-  As a matter of fact,  we are conducting the evaluation
study for them,  simultaneously assessing  its  use as a transfer standard
for our purposes.

     Mr. Stekoll:   When  you say that the  Pacific ion electrodes have not
been characterized,  what do you mean?  Don't  the companies characterize
them?
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     Mr. Basileo;  Yes.  Companies characterize most instruments and
they publish the specifications.  It has been our experience over the
years, in testing instrumentation, that some 80% to 90% of instruments
that come from the manufacturer do not meet the manufacturer's specifi-
cations.  Another high percentage do not work when we get them.

     From the floor:  You said that your target is September 1978 for
the development of standard analytical methods for nutrient heavy
metals, molecular weight hydrocarbons, plus others.  Would these be EPA/
NOAA standard procedures?

     Mr. Basileo:  Yes.  We are comparing the oceanographer's method to
the EPA's methods.  We should end up with a good set of procedures for
salt water.

     From the floor;  My reservations deal with how you are going to
prove the accuracy of the methodology.  Just about everyone does his
own procedure in the oceanography field.

     Mr. Basileo:  In some of the areas, for example, heavy metals,
there are no corresponding methodologies in an oceanographer's bible,
so this is basically a comparison of two EPA methods.

     From the floor;  In the fresh water area, the American Public
Health Association produces standard methods for water and waste water,
and there are organizations such as ASTM, which is a standard-setting
organization.  Before they adopt a method as a standard, they do under-
take rather rigorous testing procedures to determine its precision and
accuracy.  I question whether a contractor is capable in all these anal-
yses, based on a comparison of one method to another, to derive standard
procedures for EPA and NOAA.

     Mr. Basileo:  The outcome of this contract is going to be a compar-
ison of methods:  it will not automatically be a standard procedure.
We are looking for a good set of procedures for ocean water.

     Dr.Lefcort:  There is a severe need for standardization for many
of those parameters.  If EPA/NOAA were to come out with a standards
methods manual, it would be used internationally.  However, unless there
are some excellent precision and accuracy studies performed on each
method and interlaboratory studies and round-robin testing procedures
establish these variances, I did not want to see them come out as offi-
cial procedures at this time.  And I do not see what a comparison of
two methods by one contractor is going to tell us.  Where do we go from
there?

     Mr. Basileo:  I agree that the output of this would certainly not
be accepted by a lot of people until more work has been done on it.  The
outcome of this should give a set of methods to start with.  Granted,
they will not be accepted standards, but they should be a set of methods
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so that the detailed precision analysis,  and the  necessary  inter-
calibration for developing  the final,  accepted standard method  can
be done.

     Dr. Lefcort;  Well,  EPA-Cincinnati,  has the  Analytical Reference
Service, which sends samples to approximately 80  laboratories.   Based
on their results,  they calculate the precision and accuracy of  the meth-
ods and publish  it as  an  Analytical Reference Service Report, which, in
turn, is adopted by other standards-setting organizations.

     I believe that this  is very necessary in the case of sea water
analysis.   The problem with the Cincinnati Laboratory is that they have
not focused on sea water  analysis and they are badly underfunded.  The
future of  funding  for  this type of work does not  look too promising.

     Dr. Barnes;   How  can you possibly come up with a standard  method
when there are no  standards?  The' second question is, how did you choose
the EPA flame atomic  absorption and graphite furnace methods as the
potential  methods  to  use.  If you look at the data from those methods,
which  is in the  EPA book, or the data which is published from all the
laboratories, it is bad.   There is no way you can use those methods.
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              DEVELOPMENT OF OCEAN OIL SPILL CONCENTRATION
                    AND TRAJECTORY FORECAST METHODS

                            Celso S. Barrientos*
     The demand for energy and raw materials in the United States con-
tinues to increase.  Petroleum and its by-products are the main source
of energy and materials.  This ever-increasing demand has led to a sub-
stantial increase in the scale of production and transportation of oil.
Subsequently, this led to the increase in the level of pollution by oil,
especially in the marine environment where much of the production and
transportation are occurring.

     Oil pollution in the ocean and the coastal and inland waters is
largely a result of natural  seepage, continuous seepage during produc-
tion, loading and unloading  operations in port areas, the input of
industrial wastes, tanker cleaning discharge in the open sea, and major
disasters in marine transportation and offshore production.  Disastrous
oil spills occur in a more concentrated and spectacular form, often
causing severe and noticeable environmental problems.  Although disas-
trous spills have drawn much public attention, the extent of oil pollu-
tion in the "marine environment is much greater than that caused by the
much-publicized major accidents.

     The most noticeable damage to the marine environment occurs in the
coastal areas.  Wildlife and sea life are very susceptible to oil pollu-
tion damage.  Aesthetic damage to the shorelines is generally severe.
The populace is heavily affected by the loss of recreational areas, and
the fishing industry and food sources feel the impact.

     Although contingency plans to prevent damage by oil pollution have
been devised, there is an urgent need to develop methods to forecast
movements and the behavior of the pollutant.  In the case of oil spills,
viable methods are required  to predict the oil movement.

     The objective of this program is to develop a numerical model to
predict the concentrations and trajectories of petroleum in the open
ocean (not in bays or estuaries).  Concentration will be forecast as a
function of space and time.  The proposed model will cover various kinds
of sources, i.e., continuous point leakage, individual spill, or spill
*    National Weather  Service,  National Oceanographic and
Atmospheric Administration;  Silver Springs,  Maryland  20910.
                                 271

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added to a general background level of known variable concentrations.
The model will include wind, wave, and current forecasts on a small
scale in time and space not ordinarily included in the present atmos-
pheric weather forecast system.   Forecasting techniques will be devel-
oped for these variables.  -Additionally, trajectory models will require
as input oceanic variables, including depth, coastal geometry, and
physical properties of the water.  These oceanic variables will be used
in the development.

     Petroleum concentration forecasts by this model will be used pri-
marily by the National Weather Service  (NWS) in operational and research
modes.  The operational mode will be used in real time to augment sup-
port of the provisions of the national oil and hazardous substances
pollution contingency plan.*  In the research mode, potential spills
will be evaluated parametrically as in the preparation of environmental
baselines to aid decision makers in assessing potential impact of off-
shore development.  A wide range of ultimate users will potentially
receive these data.  These users include U.S. Coast Guard, Office of
Emergency Preparedness, Environmental Protection Agency, U.S. Geological
Survey, Bureau of Land Management, and National Marine Fisheries Service.
PROCESSES AFFECTING SPILLED OIL IN THE OCEAN

     Four broad classes of processes affect spilled oil:  (1) weathering,
 (2) spreading,  (3) diffusion, and  (4) transport.  These processes are
interrelated and interactive.  They are acting all the time, but some
are more dominant, depending on the time and size of the spill.
      _**
     Weathering includes processes related to the physical and chemical
characteristics of the oil, such as evaporation, coagulation, sedimenta-
tion, dissolution, emulsification, and microbial degradation.  Some of
the weathering processes are dependent on the physical variables of the
environment, e.g., evaporation is a function of temperature and wind
speed.  Weathering processes are ongoing at various stages of the spilled
oil; some are more dominant than others at those stages.

     Spreading refers to the physical expansion of the lateral size of
the spill.  Aside from turbulent processes, gravity and surface tension
are the main spreading forces.  Gravity acts through local oil thickness
*    The Department, of Commerce, through NOAA, provides support:
"...with respect to the marine environmental data; living marine
resources; current and predicted meteorological, hydrological and
oceanographic conditions for the high seas, coastal and inland waters,
including tides and currents for coastal and territorial waters and the
Great Lakes."  National Oil and Hazardous Substances Pollution Contin-
gency Plan, Council on Environmental Quality, August 13, 1973, 38(155),
Part II.
                                272

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gradients.  Surface tension,  which acts on the periphery of  the oil,
depends upon the chemical..and physical properties of the oil and the
sea water with which  it  is  in contact.  Drag on the bottom of the oil
retards expansion.  Spreading,  which usually occurs simultaneously with
diffusion and transport,  interacts with these processes.

     Diffusion is the process by which the size of the  spill grows by
random motion of the  oil and  water.   Diffusion refers here to turbulent
diffusion, which is several orders of magnitude larger  than  molecular
diffusion.  Diffusion can be  handled in the problem by  the use of gradi-
ents and eddy diffusion  coefficients.   There are estimates of diffusion
coefficients for spreading  of oil, dyes, and other conservative sub-
stances in the water.  However, the diffusion coefficient for oil may
be influenced by spreading  as well as by wind fluctuations.

     Transport refers to the  advective movement of oil  as it is being
carried by water and  wind,  when moved by water, we refer to it as motion
due to current and when  by  wind we refer to it as wind  drift.  Water
current is induced by density difference, wind, waves,  and tide.  Wind
drift is the actual action  of the wind on the oil.   The transport is
the vector sum of the currents and wind drift.   When oil is  spilled, a
certain time will be  reached  when the transport components are probably
the most dominant part of the spreading process.

     The greatest environmental problem in forecasting  oil spill trajec-
tories is the prediction of transport processes.  The interaction of the
ocean and atmosphere  is  most  important in dealing with  this  meteorolog-
ical problem.
MODEL DEVELOPMENT

     The  general scope of the program is the  development of a comprehen-
sive model  to predict the dispersion and' concentration of contaminants
from available oceanographic and meteorological  data, including informa-
tion on the winds  from a numerical weather prediction model operated at
the National Meteorological Center, NWS; the  wave spectrum as determined
from numerical models and available observations; bottom topography from
available charts;  and tidal elevations and currents.  The model winds
will be used to compute local winds that, in  turn, will cause currents
and will  move floating pollutants directly through drag.  Transport from
the wave  field will be considered as another  potential dispersal mecha-
nism.  Tidal currents as estimated by the National Ocean Survey tabula-
tions are also important.  Bottom topography  is  needed in several of the
computations.  A description of bottom topography (and coastlines) must
be provided for all coastal regions to be considered.  Plans are to con-
centrate  first on  the northeast coastal areas of the United States as a
case study  and, if this approach proves feasible and desirable, to ex-
pand the  capability nationwide.
                                 273

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     This program is interrelated with and depends on a number of on-
going activities in National Oceanographic and Atmospheric Administra-
tion (NOAA).  Specifically, statistical correlation techniques for
estimating local surface winds from model winds have been developed and
tested for several coastal locations.  Wave spectra are now regularly
measured on NOAA data buoys and waves forecast by a numerical model.
The wave program will be improved to allow for wave spectra.  Bathy-
metric surveys and their resulting charts are presently a major task of
the National Ocean Survey, as are the computations of tidal height and
current for most of the U.S. coastline.

     The Deepwater Ports Act requires NOAA to make "adjacent state"
determinations.  The Deepwater Ports Project Office (DPPO)  was estab-
lished in NOAA to perform the functions required by the Act.  Trajectory
modeling is a primary function of DPPO.  Close coordination between this
EPA program and DPPO will be maintained.   Recently, DPPO has been made
part of NOAA's Environmental Data Service.

     NOAA's Environmental Research Laboratories (ERL)  are involved in
environmental assessment of the Alaskan Continental Shelf for the Bureau
of Land Management of the Interior Department.  A major portion of this
project involves ocean circulation and trajectory modeling.  This pro-
gram will apply the relevant results from the ERL's project.

     Our program is divided into five tasks:  weathering, spreading,
diffusion, transport, and model integration.

     The first four tasks are the processes group, and the fifth, model
integration, includes management, coordination, and implementation of
the program.  The factors affecting the spilled oil will be treated
separately either by contracts or in-house work, and the results will
be integrated in an operational model in NWS.

     We summarize below the activities involved in each of these
tasks.

     Weathering

     The problems of weathering are complex.  Some of the weathering
processes are also environmental problems; e.g., evaporation is a mete-
orological problem.   Oil chemists have undertaken a lot of work on
weathering; however, this work has occurred mainly in the laboratory,
and its application to the actual marine environment is not known.  In-
stead of developing new solutions, we plan to make an extensive litera-
ture survey of existing techniques.  We will attempt to define the
various rates of the weathering processes and include those that are im<--
portant.
                                274

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     Spreading

     The spreading process is dependent  on  the buoyant and viscous prop-
erties of the oil.   Where possible,  we will apply  the pioneering study
on spreading by  James  Fay of Massachusetts  Institute of Technology.  We
believe that there is  knowledge of many  processes  affecting the spilled
oil, and interactions  with weathering, diffusion,  and transport may be
significant.

     Diffusion

     The diffusion process can be handled in two ways; by using classi-
cal eddy diffusion coefficients,  or  by introducing random fluctuations
into the equations of  motion.   To some general degree of accuracy, tur-
bulent coefficients  are known for passive tracers, such as neutrally
buoyant dyes, but the  problem is how this information can be used for
oil.  The interface  between oil and  sea  water will modify the effects
of turbulent water motions on the oil; thus,  the coefficients must be
altered.  Use of the other approach,  however, requires a knowledge of
the turbulent spectrum of water motions.  Its advantage is that both
spreading and diffusion can be combined  into one equation; thus, inter-
actions can be considered.   We plan  to investigate both approaches,
study the data,  and  select the more  useful  method.

     Transport

     Transport refers  to the interaction between oil and water movement.
The main problem here  is one of coupling:  will the oil move as the
water moves?  Most of  our development efforts will be concentrated in
this important task.  We believe that, in determining oil spill trajec-
tory and concentration, transport will be the major mechanism.  We sub-
divided this task into four subtasks: large-scale or semipermanent
current, tidal current, wave-induced current, and  wind drift current.

     Semipermanent current.  We plan to  use current tabulations from
charts or run large-scale ocean circulation models.

     Tidal  current.  Tidal current is computed or  measured for various
coastal locations.   Tidal current, which is strongly dependent on the
location, is significant in the coastal  regions and when the time scale
is less than the tidal cycle.  In the open  ocean,  it is small and may
average out to zero  when time is larger  than the tidal cycle.

     Wave-induced current.  Wave prediction models are dependent on the
wind input.  There are two methods to predict or specify waves:  singu-
lar method  (significant wave height)  and spectrum  method.  There is no
agreement yet on a unified theory of wave transport.  Simple wave trans-
port solution is that  of Stokes.   However,  surface wave-induced current
is still not determined.  We plan to do  a great deal of development in
this subtask, using  field and laboratory data and  existing models and
theories.

                                 275

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     Wind drift current.  Oil on the water surface will be acted upon
by the wind stress.  Thus, oil will skid on the water surface because
of wind action.  When the oil is absent, wind stress on the water sur-
face induces surface current.  There are two basic problems involved:
(1) prediction of surface wind at suitable time and space scales for
oil trajectory; and  (2) understanding the mechanism for the transfer of
energy from the wind to the oil or water.  We plan extensive develop-
ments on the specification and prediction of surface wind and studies
on boundary layer dynamics.  Work on this subtask will be done in-house
and by contract.

     Model integration.  The management part of the project will fall
on this task.  The program will be performed in the Techniques Develop-
ment Laboratory of NWS, which will coordinate within NOAA and with
other agencies on related studies.t This laboratory reviews research
proposals and makes  recommendations on funding.  It monitors contracts
and will implement the  trajectory model in operational mode in NWS.
ACCOMPLISHMENTS AND STATUS

      In this  section, we discuss the accomplishments and status of the
work  for each of the tasks.

      Weathering

      We have  begun a literature survey related to oil weathering in the
ocean.  Dr. Anand Apte, a National Research Council Post Doctoral Asso-
ciate, began  this survey.  We  are continuing to do some of the weathering
work  in-house.  For example, we are doing some numerical experiments on
evaporation.

      Dr. James Mattson of the  Center for Environmental Design and Data
Analysis,  Environmental Data Service, NOAA, is developing a weathering
program subroutine for NOAA's  work for the Bureau of Land Management.
We  are following up the progress of Dr. Mattson, and we will incorporate
those results that are applicable in our model.

      Dr. Ronald Kolpack of the University of Southern California has
compiled a two-volume glossary on the fates of oil in the ocean for the
petroleum  industry.  We will be using Dr. Kolpack's suggestions in our
model implementation.

      We have  recommended support of the work of Dr. Christopher Roach on
a system model to determine the fate of oil in the marine environment
 (FOME).  Dr.  Roach will do the work at Princeton University with Dr.
William Gray  (see Transport).  The FOME model assesses the relative
rates of the  various processes affecting the spilled oil.
                                 276

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     Spreading

     In our forecast manual,* we  described  Fay's  experiments and spread-
ing parameters, which are plotted to  give the  forecasters knowledge of
the slick size for an instantaneous oil  spill.  This approach neglects
the effects of weathering and other forces,  like  wind stress, on the oil.
Our experiments are continuing  to improve the  spreading model.

     We developed a numerical model of oil  spread based on the radial
equations of motion and mass.   Although  the model was not entirely
successful  (it did not properly conserve mass), it  showed that period-
ically varying water velocities (such as those that occur in wave trains)
have only a minor effect on the rate  of  spread.   For actual conditions,
however, a radial model has limited use.  We also compared the numerical
computations with analytic  solutions.

     We are programming and testing a generalized two-dimensional
spreading model utilizing point-source spills  under the action of winds
and water currents.  We plan to use the  Argo Merchant spill information
to refine the results.  One problem will be to include surface tension
in the formulation.

     Diffusion

     Dr. Gabriel Csanady of Woods Hole Oceanographic Institution has a
contract on the parameterization  of turbulent  transfer in the mixed
layer.  Dr. Csanady, who has completed the  scaling  analysis for the
bottom boundar*y layer, obtained an elegant  formulation to compute effec-
tive eddy viscosity.  The  following two  paragraphs  are excerpted from
the contractor's quarterly  progress report:

     "Original data of Lofquist (1960) have been  obtained and
     analyzed  in detail with a  view to evaluating them from the
     point  of view of modern turbulence  theory.   This work
     revealed  some unexpected properties of turbulent flow
     along  a density interface, notably  a momentum  transfer
     mechanism involving viscous  wavelets,  which  enhance
     vertical momentum  flux associated with the mean shear.
     The effective viscosity of such  an  interface has been
     successfully ^parameterized.   Further analysis, making
     use of the  "inner" and "outer" law  framework of turbulent
     boundary  layer theory resulted in the  parameterization
     of interfacial stress. Work is  in  progress  in relating
     these  results to  1972  Lake Huron observations  at the
     bottom of the mixed  layer.
     See Model Integration.
                                  277

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     Somewhat analogous results have been obtained by a re-
     analysis of data of Wu (1975) on flow underneath the air-
     sea interface.  The effective viscosity was found to be
     related to surface tension, but the exact method of param-
     eterization remains elusive.  Work is in progress on this
     problem as well as on the free surface velocity law."

     We have awarded a contract to Dr. Richard Miksad of the Civil
Engineering Department, University of Texas at Austin, on the study of
turbulent dispersion motions in both atmosphere and sea.  These motions
act to diffuse and disperse the slick.  The study will deal with air-
sea coupling in generating horizontal turbulent spreading and assess
its relevance to oil slick spreading.  Some simple laboratory experiments
will be made to determine the validity of the theoretical results.

     Transport

     At NWS, we have implemented wind forecast equations for eight
coastal stations along the east coast and six along the west coast of
the United States.  We are monitoring the forecasts and improving the
forecast equations.  A paper on "Single Station Marine Weather Forecasts
Based on Model Output Statistics" by David M. Feit was presented and was
published in the Preprint Volume:  Conference on Coastal Meteorology
sponsored by the American Meteorological Society.  This paper describes
the development procedure, implementation, and verification of the
operational wind forecast techniques for the 14 coastal stations.  We
have begun developing wind forecast equations for 40 additional stations
along the east, gulf, and west coasts, and the Alaskan region.

     We awarded a contract to the Institute of Marine and Atmospheric
Sciences, City University of New York (CUNY), on "Specification and
Prediction of the Vector Wind Stress and Wind Drift on the United States
Continental Shelf for Application to an Oil Slick Trajectory Forecast
Program."  Dr. Vincent Cardone is the principal investigator.  CUNY will
develop techniques to specify, forecast, and update the driving force
for the dominant transport mechanism in an oil slick trajectory forecast
model, i.e., the vector wind stress at the air-sea interface.  The tech-
niques developed will be oriented to operational applications.

     We awarded a contract to the Atmospheric Science Group, University
of Texas, on "Oil Spill Behavior in the Open Ocean."  The contract is
divided into three parts:  (1) large-scale atmospheric motion field
(Professor K.H. Jehn); (2) atmospheric boundary layer characteristics
(Dr. N.K. Wagner); and (3) turbulent dispersive motions in both atmos-
phere and sea  (Dr. R.W. Miksad, see Task 3).

     Professor Jehn will determine offshore winds from the more dense
observations at the coastal areas by developing a weather map typing
procedure.  Dr. Wagner will develop a method to transfer the low-level
wind field to the ocean surface through boundary layer dynamics.
                                278

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     We have recommended  funding the' proposed research by  R.M.  Parsons
Laboratory for Water  Resources and Hydrodynamics,  Massachusetts Institute
of Technology, on  "Development of a Weighted Residual Scheme  to Calculate
the 3-D Structure  of  Currents in Coastal Areas Using a Depth  Varying
Eddy Viscosity."   Professor Bryan Pearce,  the principal investigator,
will formulate a Galerkin-based finite-element numerical scheme to cal-
culate the three-dimensional velocity field applicable to  coastal areas
and inland water bodies with forcings by winds,  surface elevation, and
Coriolis force.  The  model  will be used to investigate the  functional
form of the vertical  eddy viscosity.

     We have recommended  support of the proposal from the Department of
Civil Engineering,  Princeton University,  on "The Development  of a Com-
puter Model for the Simulation of Flow and Oil Slick Movement in the
Coastal Zone" with Dr. William Gray as the principal investigator.
Dr. Gray will be assisted by Dr.  Christopher Roach (see Task  1).
Princeton University  is proposing to (1)  develop a finite element method
of solution to the complete hydrodynamic equations for the  flow of oil;
 (2) develop a computer program to determine the  fate of oil in  the
marine environment (FOME, Task 1);  and (3)  couple  the hydrodynamic flow
model with the FOME model to arrive at a computer  model for ocean oil
spill.  The proposed  model  could be used as the  core program  for the
model integration  task of the project.

     We have recommended  funding the proposal from College of Engineering,
Rutgers, the State University of New Jersey,  on  "Numerical Evaluation
of the Effect of Ocean Surface Waves on Oil Spill."   Dr. Yung Y. Chao,
associate research professor, would be the principal investigator.  The
proposed work consists of three steps:  (1)  formulation of relevant
equations on ocean surface  waves; (2)  calculation  of Stokes velocities
for various input  conditions; and (3)  development  of a numerical method
to calculate the variation  of wave drift velocity  at the surface caused
by viscous damping of oil film.  The result of this  study will  give us
estimates of wave-induced transport.

     Model Integration

     We completed  the first draft of a forecasting manual for oil spill
trajectories.  The manual includes discussions of  the four major proc-
esses affecting spilled oil and practical  forecasting techniques.  The
draft was reviewed by the NWS Western Region Headquarters with  favorable
comments.  It was  also reviewed by personnel in  NOAA's Environmental
Monitoring and Predictions  Office,  who made suggestions for improving
the manual.  We are revising the manual based on these suggestions.

     We are cooperating with other scientists working on the  New York
Bight data and developing a meteorological analysis technique to derive
the forcings for oceanographic models.  This input is needed  for the
oil spill trajectory  model.  In addition,  we are processing meteorolog-
ical data from December 15, 1976, in connection  with the hindcasting
for the Argo Merchant tanker oil spill accident.   The data  will be used
to test the model.

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SUMMARY

     We have indicated some of the reasons for the development of ocean
oil spill concentration and trajectory forecast methods.   Exhibit 1
illustrates the elements in development of an operational ocean oil
spill trajectory model and summarizes the important factors determining
the fate of oil in the ocean.*  There are four main components of the
development:  software development, and the three main subject areas
of weathering, diffusion, and transport.  The spreading algorithm is
combined with diffusion because of the close interaction between the
two processes.  We will implement diffusion by solving the dynamical
equations of two-dimensional oil spreading, using the turbulent nature
of wind and water currents to define or modify the apparent diffusion.
Weathering and transport are to be implicitly included in the expansion
of the spill.

     Total transport will be the combined output of the various water
current models  (with interaction when appropriate).  Initially, values
for some of  these inputs are expected to be from simplified approxima-
tions  (such  as a wind-driven current proportional to wind speed), but
as our knowledge and model development advance, these will be improved.
Wave transport will also influence the spreading of the spill.  An ice
model is necessary in the Alaskan area.

     Software development will address and follow the physical and
model studies and data gathering, and will dominate the final phase of
model integration.

     Weather will modify local oil properties (e.g., density, composi-
tion, state) to indicate limiting times and areas of contaminants.
Winds, temperature, and other atmospheric properties from NWS models
will be readily adaptable.

     A major part of our progress to date has been in the spreading
processes.  Our major future task is analyses of transport, which will
center on wind-drift currents.  We have NWS input of coastal winds and
are sponsoring work on air-sea interaction and turbulent transports in
the surface layer.  All these complementary "on-line" efforts are sub-
servient to the final goal of implementing a forecast procedure.  The
forecast model will be continually updated as our knowledge and abili-
ties increase.
*    The diagram is based on the one originated by Dr. Jerry Gait of
Pacific Marine Environmental Laboratory, NOAA.
                                 280

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               EXHIBIT 1:   DEVELOPMENT  OF  OPERATIONAL OCEAN
                         OIL SPILL  TRAJECTORY MODEL
Bathymetry
   And
Geography
Stochastic
Current
Analysis


Diffusion
Subroutine

                                                                            Evaporation
                                                                            Sedimentation
                                                                            Coagulation
                                                                            Oxidation
                                                                            Emulsification
                                                                            Biodegradation
                                                               MOS Wind Forecast
                                                               Fine-mesh Meteorological
                                                                     Model
                                                               Wind Specification
                                                               Surface Stress
                                                                     Formulation
                                            281

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     We are also developing the following subprograms:

     A weathering subroutine with the quantitative rates of the
     processes as a function of environmental parameters

     A spreading subroutine, or a combined spreading-diffusion
     subroutine, depending on results of our numerical  experiments

     A regional meteorological model for analysis and prediction
     of winds and stability conditions in the boundary  layer

     A coastal circulation model to compute currents, including
     wind drift

     An algorithm to compute transport from wave data

     An integrated oil spill trajectory model including all the
     above subrout ine s.
DISCUSSION

     From the floor;  Do you have any responsibility for analyzing
specific spills when they occur?

     Dr. Barrientos:  It is not clear right now.  The last time, in
New England, we supported the Coast Guard with meteorological observa-
tions.  Once this model is developed, we hope to give them more than
the meteorological probabilities.

     Dr. Baumgartner:   Do you intend to provide a service that someone
can call when a spill occurs to find out where it is going?

     Dr. Barrientos:  Yes.  Ultimately, that is what we want.  That is
why we will be needing all the geography in all the coastal areas of
the United States and Alaska, so that, at any time, we can just crank
up the model for the specific area.  The model will be available on
the NOAA computer and will have remote terminals all over the country.
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                  THE UNDERWAY WATER SAMPLING SYSTEM

                    C.D. Kearse,* R. New,t and D. Enabnit*
     Many of the open ocean phenomena  that have been the subject of
oceanographic studies in the past  can  be  characterized by a few data
points widely separated in space and time.  For example, a salinity
change of a few parts per thousand over hundreds of kilometers of open
ocean would be considered large, and the  important time scales for mix-
ing are several months or more.

     With the rise  in importance over  the last decade of environmental
management, the study of coastal and estuarine waters has increased.
In contrast to the  deep ocean, the spatial and temporal scales are small
in the shallow water areas.  In an estuary, salinity can change from the
marine value of 35  parts per thousand  to  the fresh water value of 0
parts per thousand  in 10 km, and significant mixing caused by the semi-
diurnal tides can take place in 12 hr.  In addition, many of the partic-
ular phenomena presenting problems in  environmental management, such as
pollution events, have temporal and spatial scales significantly smaller
than ocean phenomena.

     Traditional sampling methods  and  instrumentation are often ineffec-
tive in investigating adequately this  new family of smaller scale, more
transient phenomena.  Because pollution events like oil spills and barge
dumps are of short  duration, there is  insufficient time to stop, collect
a sample, characterize the water mass, and move on before the phenomenon
has changed significantly.  In addition,  there may be a net advection of
a pollution plume during sampling, which  makes it difficult to execute
a predetermined sampling strategy. Because pollutant gradients can be
large and plumes small, more accurately located and closely spaced sam-
ple points are required for adequate characterization.  The increase in
the total amount of data to be collected  can saturate existing instru-
ment systems and become expensive  to collect.
*    Engineering Development Laboratory,  Office of Marine Technology,
National Ocean  Survey,  National  and Atmospheric Administration,
Riverdale, Maryland  20845.

t    Now with the Systems  Analysis  Division,  Office of Marine
Technology.
                                283

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NJ
CO
                                                                      On Board:  Data processing and
                                                                      display, navigation subsystem, water
                                                                      sample processing and analysis
                                                                      equipment (hydrocarbons, metals,
                                                                      suspended particles, particulates,
                                                                      surface wind velocity)
Doppler
sonar
current
sensor
Umbilical
  containing
   electrical cable
                                                                                   CTD and
                                                                                   water
                                                                                   sampler
                                                                                      Optical
                                                                                      scattering caused
                                                                                      by suspended particles
                                                                              Acoustic backscattering
                                                                              caused by suspended particles
                                            EXHIBIT  1:    A  CONCEPTUAL  UNDERWAY  WATER  SAMPLING SYSTEM

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     Recognizing that  the  developed technology  was  seriously lagging,
the Engineering Development Laboratory of the National Ocean Survey,
National Oceanographic and Atmospheric Administration  (NOAA), proposed
developing an underway, water sampling system (UWSS) to aid in charac-
terizing and monitoring the regions subjected to  energy-related pollu-
tion.  This UWSS is  a  complete shipboard instrument system designed to
acquire, measure,  display, and store oceanographic, meteorological, and
navigation data while  the  ship is continuously  underway.  The Engineer-
ing Development Laboratory plans to design,  develop, fabricate an engi-
neering model, test, and develop techniques  for the use of UWSS.

     The system has  the following major components:  a water measurement
subsystem  (a towed sensor  array or instrumented undulating fish plus a
water pumping system)  designed to collect oceanographic data and water
samples; a navigation, data processing, and  display subsystem for navi-
gation, data handling, real-time processing, and  display; and a current/
depth measurement  subsystem to remotely measure ocean currents.
Exhibit 1 illustrates  this system.

     This paper  summarizes the efforts to date, describes the present
status of individual tasks, and discusses the future of the project.
The  five tasks that  are in progress or completed  are:

1.   Applications  study

2.   Navigation,  data processing, and display subsystem

3.   Current/depth measurement subsystem

4.   Overview of optical measurement techniques in  oceanography

5.   Shipboard  chemical measurement techniques.


APPLICATIONS  STUDY

     The  specification of which parameters to measure, their ranges of
values, precisions and accuracies, mean time between failure, sampling
rate,  etc. ,  should be derived from the application  in which the system
will be used.   But the UWSS is an anticipatory  system designed to be
ready  to  use when a targeted program begins  and not 3 years later.  Un-
fortunately,  the NOAA program for the monitoring  and characterization of
energy-related  pollution in which the UWSS will be  used is still in its
very early stages of formulation, making it  impossible to designate the
ultimate  application.   To circumvent this problem,  an  applications study
was  performed according to accepted methods  of  systems analysis.  This
study  was  designed to:

   Devise and describe in detail a number of  applications of a UWSS
     that  are pertinent to an understanding of  the  effects of energy-
     related pollutants in the marine environment.


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    Determine whether a "defensible rationale" exists for a UWSS.

          If the finding is affirmative, then identify what parameters
          should be sampled and the detailed requirements for sampling.

          If the finding is negative, then recommend either the speedy
          termination of the project or the extent and details of a
          follow-on program.

     The study's approach was to have detailed "scenarios" written by
oceanographic and environmental experts- describing a baseline, monitor-
ing, or special study effort pertaining to energy-related pollution.
Each of the eleven scenarios (see Exhibit 2)  used a shipborne UWSS as
the primary data collection instrument.  The major sections of the sce-
narios described a geographical site, the pollution problem, a mathe-
matical model or data utilization plan, a detailed experimental plan
including justification of stated requirements, and a concluding dis-
cussion on the advantages and limitations of using an underway system
for that application.  Appendix 1 contains summaries of five of these
scenarios.*  In addition to the scenarios, interviews were conducted
with oceanographers currently using first-generation underway systems,
and their experiences were incorporated into the report.

     By selecting a representative set of pollution problems for the
scenarios, we produced sample applications that should be similar to
those that will be required by the emerging NOAA program.  As a result
of analyzing these scenarios, we have drawn the following major conclu-
sions :

    There are a number of potentially significant applications in the
     study of energy-related pollution in which a UWSS would be extremely
     useful if not essential.

    Underway data collection opens a new regime of spatial and temporal
     scales to investigation.

    An elementary UWSS that would be useful in many applications can
     be defined from the common requirements of the scenarios (position,
     time, salinity, temperature, depth, current, real-time data anal-
     ysis and display, and the collection of a clean water sample).'

    For routine baseline and monitoring programs, there are potential
     cost savings as a result of the reduced time required for data
     collection and the automation of sample and data handling.

    Certain sensors and instruments (particularly chemical) may require
     further development before they will be usable at the higher sam-
     pling rates of a UWSS.
     Complete scenarios are available on request.


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EXHIBIT  2:    SCENARIOS  OF ENERGY-RELATED  POLLUTION
   Dr. Hosein Fallah S, Dr. Robert Stark,  Mathematica, and College of
   Marine Studies, University of Delaware, respectively.
   Oil Spill Trajectory:  Follow an oil spill to determine  the  relationship
   among wind, current, and spill trajectory.

   Dr. Keith Stolzenbach  & Dr. Donald Harleman:  Department of  Civil
   Engineering, Massachusetts Institute of Technology.
   Thermal Plumes:  Synoptically map a power plant thermal  plume  in three
   dimensions .

   Dr. Erik Mollo-Christensen:  Department of Meteorology.  Massachusetts
   Institute of Technology.
   Space  and Time  Scales:  Use underway measurements together with  fixed
   data stations  to establish scales of currents.

   Dr.  James  Brooks S  Dr. William  Sacket:  Texas ASM University.
   Dissolved  Hydrocarbon  Sniffing:  Baseline determination of dissolved
   C1~C4  hydrocarbons  near oil rigs and of relationship of surface  slick
   to petroleum hydrocarbons  in the water column.

   Dr.  James  Anderson: University of Washington.
   Phytoplankton.Patchiness:  Distribution and productivity of  phytoplank-
   ton in pollution-impacted  areas.

   Dr.  Dennis Polis  &  Dr. Victor  Klemas :  College of Marine Studies,
   University of Delaware.
    Density Fronts:  Study density  fronts  and provide sea-truth for  satel-
    lites doing same.

    Dr. Douglas Segar:   NOAA/EDL.
    Trace Metals:  Perform baseline assessment  of trace metals in New York
    Bight and dedicate ship for  monitoring pollutants at dispersed loca-
    tions.

    Mr. Earl Merritt:   Earth satellite Corporation.
    Satellite Sea-Truth:  Use UWSS to  provide  sea-truth for  satellite data
    of Pacific updwellings.

    Dr  Roff McCluney:  Florida Institute of Technology.
    Optical Properties:  Monitor optical properties  in marine areas subject
    to dredging.

    Dr. James Mattson:  NOAA/Center for Experimental Design and Data Analy-

                            ac- * .^-Tface components of NCAA test
    spill in  Alaska.

    m-  Alan  Blumberg:  NOAA/GFDL  (reduced effort).
          i~~     provide boundary values for a numerical  model of New
     York Bight.
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     This applications study is essentially complete and is being pre-
pared for publication.  The next step is to take the study to NOAA oper-
ational program managers to determine how a UWSS can be exploited in
present and future programs.  The output of this step will be considered
judgment as to the specific and most valuable application(s) for under-
way systems in NOAA.  From these applications, the Engineering Develop-
ment Laboratory can detail the specific parameters to measure and, using
the additional information in the scenarios, determine a set of func-
tional specifications and proceed with the development.  Because the
ultimate usefulness of the UWSS is critically dependent on designing
for the correct application, it is essential to estimate as closely as
possible the ultimate application.  For this reason, the applications
study is the most heavily emphasized aspect of the UWSS project at pres-
ent.
NAVIGATION, DATA PROCESSING, AND
DISPLAY SUBSYSTEM	

     Because some of the major subsystems will require long development
time, it is necessary to begin their development before final definition
of the total system.  The danger in this procedure is that these subsys-
tems may not be required in the ultimate application.  However, in our
best judgment, the navigation, data processing, and display subsystem
and the current/depth measurement subsystem will be needed in any sys-
tem, so work began on them before the applications study was completed.

     The navigation, data processing, and display subsystem (see Ex-
hibit 3) provides such functions as data logging, display, real-time
data processing, and control of sensor operation and sampling rates.
It also will provide accurate vessel navigation, utilizing input from
present navigation devices, such as LORAN, OMEGA, and RAYDIST.  The
output will be raw data on magnetic tape, a summary listing of selected
raw data, a hard copy real-time navigation plot of data acquisition
tracklines, and selected processed data displayed in real time.  The
whole UWSS is to be interactively controllable by the scientist through
this subsystem.

     The system hardware is based on a PDF 11/40 and includes the PDP
11/40, an A/D, D/A converter, a digiview display, tape drive,  disk
drive, and a keyboard/printer.  The Engineering Development Laboratory
has purchased, installed, tested, and modified these items, which are
presently operational.  Funds from the Environmental Protection Agency
(EPA) have been used to augment the computer with a plotter and floating
point hardware.

     Software is also being developed.  Because the application for the
UWSS has not been designated, the software is written in basic modules
that can be assembled later to form the actual programs.  Two categories
                                288

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                              EXHIBIT  3:    UWSS FUNCTIONAL  DIAGRAM OF  HARDWARE  CONFIGURATION

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of modules have been developed and documented to date:*  a systems macro
library of links, prints, and special functions for use with the naviga-
tion software and in doing general computations; and a set of navigation
coordinate systems conversion modules to make conversions such as X-Y
geographic positions and range-range to X-Y.  A large amount of software
development still remains to be performed.
CURRENT/DEPTH MEASUREMENT SUBSYSTEM

     The goal of this subsystem is to remotely measure a vertical pro-
file of subsurface current vectors from a moving ship.  This subsystem
will operate either as a part of the shipborne UWSS or in a stand-alone
mode, a difficult but highly desirable goal.

     After the potentially applicable technologies for remote current
measurements were investigated within Engineering Development Laboratory,
a series of functional specifications were written for a current/depth
measurement subsystem, and a request for bids issued.  After completing
the procurement cycle, a contract was signed to develop, fabricate, and
test the subsystem.

     The current/depth measurement subsystem will be based on the prin-
ciple of acoustic Doppler shifts to remotely measure water currents.  An
acoustic pulse is transmitted from a hull-mounted transducer, and both
volume reverberation  (echoes from acoustic scatterers suspended in the
water) and the bottom return are detected.  If the scatterers are moving
with the current, then the Doppler shift of the volume reverberation
gives the speed of the scatterers and, hence, the current.  The bottom
return is used to remove the Doppler shift from the ship's motion and to
measure water depth.  By emitting three beams in different directions,
the current vector can be resolved.  Using a range-gating technique on
the return signals, a vertical profile of currents can be measured.  The
analysis and interpretation of these returned signals present a signif-
icant and controversial problem.

     The contract for this subsystem is in three phases:

1.   Theoretical analysis of proposed system, production of a total sys-
     tem error model, design modifications on a similar system developed
     for the French Navy, and analysis of data from a test of the French
     Navy system

2.   Hardware fabrication

3.   Test and evaluation.
     Copies are available on request.
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At preestablished  points,  progress is evaluated with continuation of the
contract dependent on satisfactory completion of the preceding work
Presently,  the  contract is in Phase 1 with the French Navy  test  still to
take place.

     With  its own  funds, the Engineering Development Laboratory  is spon-
soring an  in-house research program on the Doppler acoustic technique to
further the probability of success on the current/depth measurement sub-
system.
OVERVIEW OF  OPTICAL MEASUREMENT
TECHNIQUES IN OCEANOGRAPHY

     Early in the UWSS project, there seemed to be  a  high probability
that at least one optical sensor would be required  as a part of any
system.  To  build a base of knowledge of relevant instrumentation, we
performed a  study of state-of-the-art measurement techniques in optical
oceanography.  This information has been incorporated in a report and is
available for use when the application is designated. The report itself
is undergoing peer review in the academic community prior to release.


SHIPBOARD CHEMICAL MEASUREMENT
TECHNIQUES	

     There is a high probability that any UWSS will incorporate chemical
instrumentation.  The Engineering Development Laboratory performed
a survey of  presently used techniques of shipboard  chemical analysis.
This information will define developments in analytical instrumentation
that must be made before incorporation in a UWSS.
 FUTURE OF THE PROGRAM

      The immediate  future of the UWSS is not bright.  EPA has decided
 not to continue  funding the project.  This is unfortunate since we and
 the Office of Ocean Engineering  (the NOAA overview office for this proj-
 ect) are convinced  that the concept is of considerable merit.  Interest
 in the UWSS is also increasing elsewhere within NOAA.  Both Marine Eco-
 systems Analysis and the National Ocean Survey Ocean Dumping Program,
 major operational programs of NOAA, are interested in trying a UWSS in
 their applications.  Representatives from NESS, Center for Experimental
 Design and Data  Analysis, National Environmental Satellite Service,  and
 the Geophysical  Fluid  Dynamics Laboratory have been involved for some
 time in discussions with the Engineering Development Laboratory on how
 a UWSS could be  used in their applications and what specific require-
 ments they would have.
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     To obtain the full value of the work that has already been done,
Engineering Development Laboratory intends to continue the applications
study to its logical conclusion, i.e., a decision by NOAA management as
to the specific and most valuable NOAA application of a UWSS.  This will
involve finishing the study, meeting with major program managers and
planners, and a formal decision.

     The premature end of this program is disappointing.  Only rarely is
it recognized that scientific programs have a responsibility to continu-
ally help advance the state of the art in instrumentation and instrument
systems.  Yet if this responsibility is not met, new and more difficult
problems will be met with progressively more archaic instruments.
APPENDIX 1:  SUMMARIES OF FIVE SCENARIOS

     We summarize below five scenarios:  trace metals, dredge spoils,
biological effects, thermal plumes, and hydrocarbon sniffing.

     Trace Metals

     Many chemical contaminants are introduced to the coastal marine
ecosystem as a result of the energy-related activities of man.  The
impact of these contaminants on the biosphere is largely unknown be-
cause of the paucity of information concerning the chemical ecology
of coastal ecosystems.  In his scenario, Dr. Segar uses an UWSS, with
onboard analysis, to perform a chemical oceanographic baseline survey
of the New York Bight Apex.  The apex is a region presently and poten-
tially affected by man's activities related to energy production,
transportation, and utilization.

     A baseline survey of the bight was performed as part of NOAA's
Marine Ecosystems Analysis Program.  The program was a landmark in chem-
ical oceanography, but like all first-of-a-kind studies, posed as many
new questions as it answered.  The scenario describes how the initial
baseline assessment of New York Bight was done, develops a strategy to
perform this assessment with an underway sampling system, and demonstrates
how the use of such a system would render the assessment of much greater
value.

     The proposed, improved baseline survey of New York Bight Apex con-
sists of a minimum of three, 5-day cruises over a 1-year period with
standard trackline surveys taken over the same sampling grid on the
first, third, and fifth days.  The remaining 2 days of each survey are
reserved for detailed study of the areal extent and time history of
anomalous contaminant plumes observed during the standard distributions
of chemical parameters in the study area.  These distributions need to
be detailed (horizontal changes in concentration must be resolved over
distances of <_ 1 mile) and to be obtained within a short period  (< 24
hr)  to enable the resolution of the temporal changes of several days
that are characteristic of the New York Bight Apex.

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     in a second  task,  the onboard analysis, the underway sampling  sys-
tem 1S used to perform the continuing monitoring required in  the  New
York Bight Apex.   This  is achieved by a single repetition of  the  base-
line survey cruises  several times each year.  This monitoring strategy
would identify long-term changes in the ecosystem and frequent short-
term events, but  would require supplementing with source monitoring to
identify isolated short-term contamination.

     The chemical parameters that would be determined onboard would vary
for each affected coastal region.  The parameters could be changed as
more knowledge of the ecosystem is gained,  since some of these chemi-
cals cannot presently be analyzed rapidly aboard ship, the underway sam-
pling system must prepare and store samples for later laboratory  analy-
sis.  A detailed  list of the parameters is collated in the scenario,
which is too extensive to include here.

     Using a UWSS for the baseline survey and monitoring has  several ad-
vantages over  conventional station grid surveys with shore-based  analy-
sis:

1.   The  system provides the most cost-effective, least labor-intensive,
     and  least contamination-prone means of fully utilizing the capabil-
     ities of  the available analytical instrumentation.  Instrumented
     analysis  systems for a number of parameters are now available that
     are  capable of either analyzing a continuous sample stream or
     accepting small volume samples for analysis at a rapid rate.  Ob-
     taining water by standard hydrographic sampling and supplying dis-
     crete  samples to these instruments at a sufficient rate  to utilize
     fully  their capabilities are virtually impossible and inferior to
     a  continuous sample stream.

2.   The  temporal and/or spatial resolution of chemical concentration
     variations will be improved if the pumped sample stream  is ob-
     tained with the vessel underway and the rapid analysis capability
     utilized.  The underway system eliminates station time and,  there-
      fore,  more locations can be sampled in a given time.  Characteriza-
     tion of the chemical distribution in a given area can be completed
     more quickly and repeated more often.

3.   Events of short duration that are subject to water mass  motions
     may  be followed using the underway sampling capability and the
      interactive capability of onboard data display.  The interactive
      capability provided by the onboard analysis and data display is
      essential to optimize sampling of a dynamic anomaly.

      Dredge Spoils

     Energy-related pollutants, such as biocides from power plant heat
exchanger cleaning operations, heavy metals from shore-based  energy-
related manufacturing processes, and petroleum products have  been found
to accumulate in bottom sediments.  Short-term resuspension of several
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layers of this polluted sediment can result in toxic concentrations
of these materials in the water column.  Through these mechanisms,
dredging operations can have a strong impact on the introduction and
distribution of energy-related pollutants in coastal areas.  In his
scenario, Dr. McCluney uses a UWSS to determine the generation and
transport of particulates caused by dredging in Tampa Bay, Florida.

     There are additional effects of increased suspended sediment con-
centration.  Organisms such as benthic algae, other bottom vegetation,
and some marine animals are sensitive to increases in turbidity.  High
turbidity blocks out sunlight and reduces photosynthesis in bottom vege-
tation and near-bottom phytoplankton.  The siltation accompanying dredg-
ing may destroy rooted bottom vegetation, exposing bottom sediments to
wave and tidal action that can further increase the turbidity and spread
the effects of siltation.  The marine ship-channel dredging operation
planned for Tampa Bay threatens continued water quality degradation,
unless the mechanisms of particulate transport and resedimentation can
be determined and the information used to minimize the adverse conse-
quences of the project.

     Dr. McCluney will use the underway system to perform a detailed
particulate concentration survey of a dredge spoil plume.  The resulting
information will be used to develop a better understanding of the mech-
anisms of particulate transport and deposition related to dredging oper-
ations in Tampa Bay.

     The strategy will be to criss-cross the dredge plume with a series
of roughly parallel transects to monitor continuously the particulate
concentration in each size range at reselected depths.  In addition,
the wind, bottom depth, current, and salinity will be measured.  A com-
plete series of 40 transects across the plume will be completed in 1/10
the semidiurnal tidal period of 10 hr to provide adequate resolution.
A series of five consecutive surveys will be conducted over one full
tidal period.  These surveys will be repeated under varying conditions
of wind strength, wind direction, and fresh water runoff.

     Then,  a UWSS will be used to determine the areal extent of the ef-
fect of dredging on sunlight penetration.  This task is designed to take
a broader look at the effects of dredging in a large area surrounding
the dredge; the emphasis is to be upon developing a relationship between
changes in light penetrability and the concentration of dredged parti-
cles.  The particulate size and index of refraction will be used to ob-
tain an estimate of the ratio of dredged particle concentration to ambi-
ent particle concentration.  Increases in the ratio from dredging oper-
ations will be compared with changes in the penetration of sunlight to
the bottom to determine empirically any correlations that may exist
between these two parameters.
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     The underway  system will  perform a detailed particulate concentra-
te survey of the region surrounding a dredge plume.  The principal
parameters measured will be the particulate concentration in each of the
S1ze ranges and  in each of several ranges  of particulate index of refrac-
tion.  The particulate  concentrations and  irradiances will be measured
at each of a  series of  horizontal locations covering an area of approxi-
mately 100 km (more than 10 times the expected area of the visible
dredge plume).   The salinity,  current,  wind, position, depth, and time
will also be  measured.

     The strategy  will  be to criss-cross the dredge plume with a series
of roughly parallel transects  and to monitor continuously the above
parameters.   A survey of the study area will be completed in 1/10 the
semidiurnal tidal  period of 10 hr.  As  with the preceding task, these
surveys will  be  repeated at five different stages  of the semidiurnal
tidal cycle,  during varying conditions  of  wind strength, wind direc-
tion, and fresh  water runoff rate.  It  is  very important that a complete
tidal cycle series of these surveys be conducted before the start-up
of dredging operations  to provide baseline information.

     For this scenario, the underway mode  of data  collection is seen as
a technique for providing needed water quality data over large areas
with the relatively high spatial and temporal resolutions required, but
without greatly increased costs for collecting these data.

     Biological Effects

     The actual effect  of pollution on the environment is a question of
major concern.   In his  scenario, Dr. Anderson uses an underway system to
study the impact of energy-related pollution on the lower trophic levels
of the marine food chain.

     The marine food  chain consists of four pools  of living organisms
and  a pool of nutrients and solar energy.   Each pool "feeds" on the pool
beneath it  in the chain.  At the lowest level, plants feed on the nutri-
ents and solar energy.   Proceeding up the  chain, herbivores feed on the
plants; carnivores (fish) feed on the herbivores;  and man feeds on the
carnivores.   If pollution adversely affects the plankton and herbivores,
 the  supply of fish available to man can be affected  through two mecha-
nisms:   the  fish themselves pass through these  lower trophic levels as
planktonic  eggs and herbivore larvae; and  these two trophic pools serve
as a food  source for  fish.  Therefore,  the impact  of energy-related
pollutants  on these lower trophic levels is directly linked to the pro-
ductivity  of  commercial and sport fisheries.

     To identify critical interactions between the marine food chain and
energy-related pollutants, the following information must be collected:

    Biomass  distributions of the relevant trophic pools
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    Body size and life-cycle stage of the dominant species in each
     trophic pool

    Dominant species composition of each trophic pool

    Rates of biologically mediated fluxes between the trophic levels

    Current field

    Physical fluxes of the trophic pools

    Active transport by sinking and migration of the organism

    Diffusion coefficients when relevant to a trophic pool.

     This information is required in the differential equations used to
model the biological pools.  Dr. Anderson has proposed three tasks,
using an underway system to collect the above information.

     In the first task, the growth limitation of plankton caused by pol-
lution will be measured, by following an isopleth, such as the chloro-
phyll maximum, over the entire length of Puget Sound.   This must be done
in 6 hr and repeated during periods of maximum plankton growth and her-
bivore grazing.  By comparing polluted and unpolluted regions, it should
be possible to document the cell size change of plankton and the spatial
extent of the change.  This task takes advantage of the UWSS's capabil-
ity of rapidly sampling a large area such as Puget Sound to determine
the magnitude of a pollution-biota interaction.

     The immediate mortality caused by an actual, accidental oil spill
will then be determined in task 2.  This task requires that a "ribbon"
of data be collected.  A data ribbon is a line of continuous data near
the surface, a line of continuous data at a fixed depth, and a continu-
ous line of data oscillating between the upper and lower fixed data
lines.  From ribbons of current data and the plankton concentration, the
physical flux can be subtracted from the total flux leaving the biologi-
cal flux.  The biological flux can then be compared to a neighboring un-
polluted region to determine the oil-induced mortality  (which is a bio-
logical flux).  The task takes advantage of the UWSS's ability to fur-
nish a continuous sample for analysis.

     In task 3, the difference between physically and biologically medi-
ated fluxes of several parameters will be determined.   It is the biolog-
ically mediated flux that passes energy up the food chain and ultimately
determines the fishing flux.  This information can be used in the first
two tasks to distinguish physical flux effects from pollution-induced
growth and mortality.  To perform task 3, a selected parcel of water
will be continually circumnavigated and bisected.  Since the system is
underway, the same parcel of water can be measured even though it trans-
lates with the current.  From the collected data, the vertical and lat-
eral physical fluxes can be computed.  A total volume rate of change in


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the trophic pool parameters can also be computed.   The difference be-
tween the total rate  of change and the biologically mediated rate of
change is the physically, induced fluxa

     To perform the above three tasks, an underway data collection sys-
tem needs to be able  to measure:  light intensity  in the water,  chloro-
phyll fluorescence, millimeter-size particles,  temperature, salinity,
depth, current, the specific pollutants, nutrients, and dissolved oxygen.
It must also be able  to:  collect and filter a  continuous water  sample
for zooplankton; determine 14C uptake, 'election transport system activ-
ity, and ATP concentration; and identify plankton  and herbivore  species.

     The major advantages furnished by underway data collection  are:
increased data density, ability to follow a moving parcel of water, and
collection of data over a large area in a short time.

     Thermal Plumes

     One of the energy-related pollutants that  has received a large
amount of public attention is thermal pollution from power plants.
In his scenario, Dr.  Stolzenbach uses a UWSS to help analyze and
monitor the heated discharge from a large offshore electric gener-
ating station.

     Of the  10    Btu of fuel used in U.S. steam power  plants, approx-
imately two-thirds is rejected into the environment as waste heat.  When
discharged into the marine environment, this waste heat can alter the
dissolved oxygen  concentration, change the growth  rates and behavior of
aquatic organisms, and cause death in the biota from exposure to ele-
vated temperatures.   The biological effects are known  to depend on the
frequency and duration of exposure to elevated  water temperature, and
the total magnitude of the effects will depend  on  the  amount of water
affected.  Therefore, to study the impact of waste heat on the environ-
ment, it is  necessary to know the full temporal and spatial distribution
of the induced  temperature changes.

     One source of marine thermal pollution will be the Atlantic Gener-
ating Station  (ACS),  a proposed 2300 MWe floating  nuclear  power station
to be anchored  off Little Egg Inlet, New Jersey.  The  ACS will use once-
through cooling and discharge 4640 cfs of water heated to 16.1F above
the ambient.  After mixing with the receiving ocean water, the heated
discharge will  establish some farfield temperature distribution.  This
distribution has  been modeled as a differential heat conservation equa-
tion.  The first  task proposed by Dr. Stolzenbach  is to use a UWSS
to measure the  site-specific coefficients in the farfield model.

     The site-specific coefficients are the temperature diffusion coef-
ficients for all  three dimensions.  These coefficients will be mea-
sured using  a dye tracer to simulate the thermal plume.  Once the dye
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has been dispersed, a three-dimensional map of temperature, dye concen-
tration, position, and time must be collected as a series of snapshots.
The ability of an underway system to build such a map in a highly synop-
tic fashion is a new capability with wide applicability in oceanography.

     Task 2 involves measuring the actual farfield temperatures result-
ing from the ACS power plant discharge.  The objective is to construct a
three-dimensional contour map and to monitor its change with time.
Since many of the ecological impacts are a function of the duration of
exposure to elevated temperatures and the magnitude of the temperature
increase, a time-varying contour map, which allows one to describe how
often and for how long different volumes of water are subjected to what
elevated temperatures, can delineate the critically affected volumes.
To construct such a map, a system towing a string of temperature sensors
will sweep back and forth through the plume, recording temperature, po-
sition, and time.  A single, moored current meter will provide data to
remove the effects of tidal current displacement.

     Hydrocarbon Sniffing

     The production of oil from offshore wells is a source of oil pollu-
tion in the sea.  In their scenario, Drs. Brooks and Sackett use a UWSS
to determine the transport and fate of production-related oil pollution
on the Louisiana Shelf.  Two specific pollution sources are investigated:
underwater venting of the natural gas by-product of production, and
accidental spills at a production site.

     A small amount of natural gas is recovered as a by-product of oil
production.  Because it occurs in uneconomically recoverable quantities,
the gas is burned or vented underwater.  The primary components are the
C1~C4 light modular weight  (LMW) hydrocarbons, but a certain amount of
the toxic water-soluble Cc-Cj^ compounds is also vented.  The LMW hydro-
carbons are valuable short-term traces of these toxic, soluble compo-
nents of petroleum.  A system known as a hydrocarbon "sniffer" is used
to measure these hydrocarbons.  A "sniffer" is a submerged pump con-
nected by flexible hose to a gas separator, a gas chromatograph, and a
total hydrocarbon detector aboard ship.  The sniffer is towed at some
depth through an area of interest while sampling and analyzing the water
for LMW hydrocarbons.

     The sniffer system will be used to make a detailed concentration
map of the hydrocarbon plume produced by venting.  From such a map, one
can determine the dissolved hydrocarbon composition, the directions of
advection of the surface and subsurface plume, the dilution of the oil,
and the loss of dissolved hydrocarbon to the atmosphere.  The survey
tactic will be to prescribe a 2-mile square around the vent to locate
and size the plume and then to sweep back and forth across the plume at
continually greater distances from the vent.  Water will be pumped from
two depths to analyze for LMW hydrocarbons and for total hydrocarbons.
                                298

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Oceanographic parameters (salinity, temperature, dissolved oxygen,  cur-
rent, and depth)  will  be collected simultaneously with the water  sam-
ples.

     Two new advantages of an underway system have been demonstrated
here.  Since station time for sample collecting has been eliminated,
one has enough  extra time available to run a preliminary 2-mile square
around a vent.   This allows the user to optimize the rest of  the  sur-
vey by eliminating many measurements outside the plume.   A second
advantage is that sample collecting has been automated and, therefore,
is much easier  than the traditional hydrocast.

     The underway sniffer will then be used as a quick reaction system
to study the dissolved components in an accidental oil spill.  For
example, assume there  has been a platform mishap resulting in an  acci-
dental release  of oil  to the sea.  The underway sniffer  could be  de-
ployed quickly  to examine the spatial distribution and depths of  pene-
tration of  LMW  hydrocarbons as they dissolve and evaporate.   Because
the  light hydrocarbons C1-C12 evaporate quickly, about 50% of the spill
will be lost to the atmosphere in a few hours.  Thus,  the ability of the
underway sniffer to survey an area rapidly is essential.

     The same data are required in both tasks, and both  require an
interactive capability.  The underway system must be able to  take sam-
ples, analyze them, and display the results in real time so an experi-
menter can  tailor the  survey to the actual oil plume.

     This  scenario describes an underway system that is  already in  oper-
ation and  that  the authors have used.  Therefore, advantages  that Drs.
Brooks and  Sackett claim for the technique are proven advantages, i.e.;

   Speed  of operation.  The system supplies samples to the  analytical
     equipment  at the  maximum rate at which analysis can be performed.
     It also  travels from data point to data point quickly because
     station  time has  been eliminated.  As a result, the sniffer  can
     perform  in 2 hr a survey that would take 24 hr by conventional
     techniques.

   Adaptability.  Coupled with real-time data display,  the  speed  of
     operation  allows  one to alter the survey tactic in  response  to a
     changing  environment.

   convenience.  By supplying a continuous water stream, the incon-
     venience  and labor of taking hydrocasts have been eliminated.
                                 299

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UWSS Functional Diagram of Hardware Configuration
Oceanographic and
Navigation Sensors
                                                                             Scientist
                                                               Digivue Display
1
1 Versatec
1 Printer
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1

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Pen Plotter
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             SHIPBOARD ENVIRONMENTAL DATA ACQUISITION SYSTEM
                   Douglas W. Downen and Richard R. Reynolds*
      Since 1853,  the National Weather Service  (NWS) has operated a
volunteer  observing program aboard ships to provide support to marine
interests  in the  form of forecasts and warnings.  These forecasts and
warnings must be  kept updated at all times.

      Observations from data-sparse ocean areas are very important to
good  forecasts as well as timely warnings of potential problems.   With
increasing emphasis on utilization of the oceans as a source of food
and energy,  the large user community for environmental marine data is
growing.   In addition, this increased use of the oceans has brought
about a new concern about ocean dumping and pollution.

      Recently, there has been concern whether NWS receives  sufficient
observations to support their mission.  Observations are collected from
moving ships and  fixed platforms and transmitted over various communi-
cation routes. Presently, these communications are inadequate.   Many of
the observations  never reach the intended destination or arrive there
too late to  be of value to the real-time user.   Only a small percentage
of observations made by the ship's crews are actually transmitted and
delivered  to NWS  forecast offices.  Of the approximately 60,000 marine
weather observations taken each month, only 30,000 ever reach the
National Meteorological Center (NMC).   Only 9,000 of these  are  in time
to be included in the first map analysis.   Furthermore,  there are peri-
ods when nearly no data are available from certain ocean areas.

      Real-time reporting is now feasible,  and satellites in geosta-
tionary orbits can now provide a direct data link from ships to NWS.
Currently,  the NWS is developing an automated environmental data  obser-
vation system for shipboard use.   Known as the Shipboard Environmental
Data  Acquisition  System (SEAS), this  system will provide real-time moni-
toring of  meteorological and oceanographic parameters.   It  will be oper-
ated  on board various ships (e.g., oil tankers,  merchant ships),  and the
observed environmental data will be transmitted via satellite communica-
tions to NWS in Suitland,  Maryland.   The SEAS will provide  routine
*    National Weather Service,  National Oceanographic and Atmospheric
Administration,  Silver Spring,  Maryland  20910.
                                 301

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observations from relatively data-sparse oceanic areas.  Correlation
of these data with additional environmental data will result in
improved weather forecasts and warning services.

     The SEAS data and improved environmental data forecasts will be
available for a wide range of users  (e.g., coastal zone managers) to
analyze offshore exploration and resource exploitation, deepwater
port operators, and tanker routing consultants.  In addition, it will
be used as input to the oil spill concentration and trajectory fore-
cast model currently under development by the Techniques Development
Laboratory, Systems Development Office, NWS.  This model will assist
in providing more efficient clean-up operations in the event of a spill.

     Many other energy and pollution-related activities will benefit
from SEAS.  For example, with pollution clean-up operations aided, the
possibility of spills reaching coastal areas will be reduced, thus pre-
venting damage to beaches and death to ecosystems.  Improved forecasts
of heavy seas and strong winds will be used by ship-routing personnel
to steer tankers away from dangerous storm areas to avoid tanker break-
ups.  In addition, the efficiency of oil tanker operations will be
improved with better environmental forecasts.

     The SEAS program is being developed in a series of phases culmi-
nating in a fully automated meteorological/oceanographic observing
system.  The first phase is the engineering model (SEAS-1), which was
designed and built under a contract with Industrial Meteorology, Indian
Harbour Beach, Florida.  Installation of SEAS-1 on the NOAA ship
Researcher was completed in August 1976, and since that time it has
been undergoing test and evaluation.  The primary purpose of SEAS-1
is to demonstrate the practicability of using an automated system for
the acquisition and communication of environmental data in a harsh
shipboard environment.

     Exhibit 1 depicts the basic components of SEAS-1.  The wind and
temperature sensors are mounted on the mast, the data collection plat-
form radio set (DCPRS) mast electronics are on the stack, the pressure
sensor is just forward of the operations control center (OCC), and the
sea water temperature and conductivity sensors are in the bow bubble of
the Researcher.  The central terminal unit housing the display, com-
puter, keyboard, and DCPRS is in the OCC.  The keyboard is used by the
observer to input ship's position, course and speed,  and visual param-
eters (e.g., clouds, obstructions to vision, visibility).

     SEAS-1 transmits a complete marine weather observation at three-
hourly intervals, which is transmitted via the Geostationary Operational
Environmental Satellite (GOES) to the NWS Wallops Island, Virginia,
Receiving Station.  It is then transmitted to Suitland, Maryland, for
correlation with additional environmental data.  In addition, SEAS-1
displays all observed and transmitted data at the central terminal unit.
                               302

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                 WINDS
              J3ELFORT AEROVANE
            CONDUCTIVITY
              SALSSAUSH
             TOROIDAL COIL
        SEAV.'ATER TEMPERATURE
            YELLOW SPRINGS
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                  EXHIBIT 1:   SEAS-1 CONFIGURATION
      The sensors and processing system have functioned without any
 failures.  The only  problems encountered to date have been in the
 satellite communications,  and the antenna seems to be the primary
 problem.  A newly designed antenna will be tested this spring.

      An automated expendable bathythermograph  (XBT) launch system for
 SEAS  is being developed  by Plessey Environmental Systems, San Diego,
 California.  This system will be integrated with SEAS-1 in March 1977
 and undergo operational  test and evaluation on the Researcher.  The
 XBT system has the capability of automatically launching XBT probes
 from  ships cruising  at speeds up to 30 kt and providing data from
 depths  up to 760 ra.  The launch portion of the system is capable of
 storing a ready supply of  48 probes.   The need for ship personnel
 assistance will be minimized with this storage capability.  Data
 measured by the XBT  system will be processed by the SEAS-1 data pro-
 cessor,  and a standard bathythermograph report of sea water tempera-
 ture  versus depth will be  transmitted.

      To  gain experience with typical  ship operations personnel,  we plan
 to  install SEAS on a ship-of-opportunity this fall.   Since many merchant
vessels  are now using the  Maritime Satellite (MARISAT),  we may recon-
figure SEAS-1 to use this  communications medium in lieu of GOES.  How-
ever, economic considerations lean toward using GOES.
                                  303

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     The NWS envisions three  levels  of automation sophistication for
SEAS.  The first is a simple  manual  entry device, keyboard, or thumb-
wheel, interfaced with satellite  communications equipment.  This system
requires the weather observer to  input the complete marine observation.
The second level is similar to SEAS-1, requiring manual entry of ship's
position, course and speed, and visual parameters,  i.e., clouds,
weather, visibility, and  wave formation.   The third level of automation
is a fully automated SEAS.  This  design will provide a fully automated
marine observation, except for visual  parameters.

     Currently, NWS is preparing  a program development plan for SEAS,
in which the requirements for the type and number and SEAS systems will
be defined.
                                304
                                           'U.S. GOVERNMENT PRINTING OFFICE : 1977 0-720-335/6012

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