r/EPA
United States
Environmental Protection
Agency
Region 10
1200 Sixth Avenue
Seattle WA 98101
Alaska
Idaho
Oregon,,
Washington
Environmental Services Division
December 1986
Reconnaissance Survey
Of Eight Bays
In Puget Sound
Volume I
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RECONNAISSANCE SURVEY OF EIGHT BAYS
IN PUGET SOUND
Final Report
Volume 1
to
U. S. Environmental Protection Agency
Region 10
Prepared under Contract DE-AC06-76RLO 1830
and
Interagency Agreement No. DW89930272-01-1
EPA Project Officers:
Dr. G. L. O'Neal
Mr. R. R. Bauer
by
Pacific Northwest Laboratory
Battelle, Marine Research Laboratory
439 West Sequim Bay Road
Sequim, Washington 98382
Operated for the U. S. Department of Energy
by Battelle Memorial Institute
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ACKNOWLEDGMENTS
•
This final report summarizes studies conducted July, 1983, through
April, 1985, by Pacific Northwest Laboratory (PNL) Marine Research
Laboratory (MRL) for the U.S. Environmental Protection Agency Region 10,
Seattle, Washington, under Contract DE-AC06-76RLO 1830. This final report
incorporates data developed cooperatively with the U.S. Environmental
Protection Agency (EPA), Region 10 Laboratory, Manchester, Washington, and
the National Oceanic and Atmospheric Administration Northwest and Alaska
Fisheries Center, Seattle, Washington. This project was under the
administrative direction of Dr. John A. Strand, Marine Sciences Section,
Earth Sciences Department, PNL Marine Research Laboratory. The following
staff members of PNL, the U.S. Environmental Protection Agency, and the
National Oceanic and Atmospheric Administration (NOAA) were the principal
contributors to the studies:
Marine Research Laboratory, Sequim, Washington
Dr. E. A. Crecelius - Sediment Chemistry
Dr. R. E. Elston - Oyster Larval Bioassay
Mr. G. W. Fellingham - Statistics
Dr. W. H. Pearson - Benthic Infauna
U.S. Environmental Protection Agency, Region 10 Laboratory, Manchester,
Washington
Mr. R. L. Arp - Sediment Chemistry
Mr. J. M. Cummins - Amphipod Bioassay
Dr. J. N. Blazevich - Sediment Chemistry
Mr. A. R. Gahler - Sediment Chemistry
Ms. C. E. Gangmark - Amphipod Bioassay
Mr. S. V. Pope - Sediment Chemistry
Mr. R. H. Rieck - Sediment Chemistry
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National Oceanic and Atmospheric Administration, Northwest and Alaska
Fisheries Center, Seattle, Washington
Dr. D. C. Malins - Fish and Invertebrate Pathology
Dr. B. B. McCain - Fish and Invertebrate Pathology
Mr. M. S. Meyers - Fish and Invertebrate Pathology
Mr. 0. P. 01 sen - Fish and Invertebrate Pathology
We particularly wish to thank Dr. G. L. O'Neal, Mr. R. R. Bauer, and
Mr. D. L. Petke of the Environmental Services Division, and Dr. J. W.
Armstrong, Office of Puget Sound, Water Division, U.S. Environmental
Protection Agency, Region 10, Seattle, Washington, for their advice and
help throughout this project. Dr. O'Neal and subsequently Mr. Bauer
served as EPA Project Officer.
Other contributors, their affiliation, and their role on the project
were:
Marine Research Laboratory, Sequim, Washington
Mr. D. D. Andaleon
Statistics
Mr. N. S. Bloom
Sediment Chemistry
Ms. 0. A. Cotter
Sediment Chemistry
Mr. S. A. Crossman
Typist
Ms. J. M. Engel
Typist
Ms. M. L. Fleischmann
Sediment Chemistry
Ms. J. M. Kaps
Typist
Mr. S. L. Kiesser
Field Party Chief
Ms. J. N. Pfeifer
Typist
Dr. J. R. Skalski
Statistics
Mr. J. W. Sutherland
Field Party
Ms. J. A. Trelstad
Typist
Ms. M. T. Wilkinson
Benthic Infauna
Mr. P. Wilkinson
Benthic Infauna
Mr. J. Q. Word
Benthic Infauna QA/QC Review
Mr. J. S. Young
Oyster Larval Bioassay
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Pacific Northwest Laboratory, Richland, Washington
Mr. D. W. Carlile Dr. R. G. Riley
Statistics Sediment Chemistry
Ms. J. L. Downs-Berg Mr. W. L. Tempieton
Editor Advisor
Ms. S. L. English
Quality Assurance
U.S. Environmental Protection Agency,
Washington
Mr. D. J. Book
Facility and System
Maintenance
Ms. G. P. Bossier
Amphipod Bioassay
Ms. W. L. Daniels
Field Party
Mr. T. M. Davis
Facility and System
Maintenance
Ms. T. DeGaugh l
Data Management
Ms. B. J. Haynes
Typist
Mr. A. J. Hess
Field Party
Region 10, Manchester and Seattle,
Ms. K. Irby
Amphipod Bioassay
Mr. M. Matta
Research Vessel Operation
Mr. J. F. Pankanin
Field Party
Mr. W. B. Schmidt
Field Party Chief
Mr. D. A. Terpening
Field Party
Mr. W. B. Towns
Quality Assurance
Ms. T. van Woudenberg
Amphipod Bioassay
Mr. B. A. Woods
Sediment Chemistry
Mr. J. R. Yearsley
Field Party
U.S. Environmental Protection Agency, Environmental Research Laboratory,
Newport, Oregon
Ms. F. A. Cole Dr. R. C. Swartz
Statistics Amphipod Bioassay
Computer Science Corporation (contract Employee),
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The information in this document has been funded wholly by the United
States Environmental Protection Agency under contract DE-AC06-76RLO 1830
and Interagency Agreement DW89930272-01-1 with the United States
Department of Energy and Pacific Northwest Laboratory. It has been
subject to the Agency's peer and administrative review, and it has been
approved for publication as an EPA document.
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National Oceanic and Atmospheric Adminstration, Northwest and Alaska
Fisheries Center, Seattle, Washington
Mr. D. R. Bunnel Dr. M. M. Krahn
Field Party Field Party
Ms. T. K. Collier Ms. L. K. Moore
Field Party Field Party
Ms. P. K. Emry Mr. P. D. Plesha
Field Party Field Party
Mr. D. W. Gronlund Ms. L. D. Rhodes
Field Party Chief Pathologist
LTJ6 E. 6. Hawk Mr. H. R. Sanborn
Field Party Field Party
Ms. L. L. Johnson Mr. D. D. Webber
Pathologist Field Party
Washington State Department of Ecology Laboratory, Manchester, Washington
Mr. D. D. Huntamer Ms. M. D. Stinson
Sediment Chemistry Sediment Chemistry
Mr. M. H. Schlender
Sediment Chemistry
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EXECUTIVE SUMMARY
Over the past two years, Pacific Northwest Laboratory's (PNL) Marine
Research Laboratory (MRL) has conducted a research program in Puget Sound
with the objective of developing a better understanding of the toxic
contamination problems in selected urban-industrialized bays.
Two major tasks comprised this effort. First, screening surveys and
analyses were conducted in four urban-industrialized embayments
(Bellingham Bay, Port Gardner - Everett Harbor, the Fourmile Rock -
Elliott Bay dump site vicinity, Sinclair Inlet) and in four baseline
embayments (Samish Bay, Case Inlet, Dabob Bay, Sequim Bay) to serve as a
guide for more detailed technical analyses. The second task involved
conducting detailed surveys and analyses in the same bays, but at a
limited number of stations.
The results of these surveys are given in this report: a description
of each urban and baseline bay in terms of its sediment chemistry,
sediment toxicity, numerically dominant benthic infauna, and incidence of
fish and shellfish disease. Each biological parameter (benthic infauna,
incidence of fish and shellfish disease, amphipod and oyster larval
bioassay) is discussed in relation to the physical and chemical properties
of associated sediments. Present research findings are compared with
results of historical surveys. Finally, the embayments and the stations
in each embayment that currently show signs of degraded sediment quality
are identified.
The organization of this study was such that the U.S. Environmental
Protection Agency (EPA), Region 10 Laboratory; the National Oceanic and
Atmospheric Administration (NOAA), Northwest and Alaska Fisheries Center;
and Pacific Northwest Laboratory, Marine Research Laboratory participated
in developing material included in this report. The EPA, Region 10
Laboratory staff assisted in conducting screening surveys in the fall of
1983, and conducted the detailed surveys in the spring of 1984 themselves.
They also analyzed for priority pollutants and performed amphipod
bioassays. The NOAA Northwest and Alaska Fisheries Center conducted
independent surveys in the same urban-industrialized and
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undeveloped embayments to obtain fish and shellfish for analyses of
neoplasia and other diseases. The MRL assisted in conducting the
screening surveys, performed range-finding chemistry for both the
screening and detailed surveys, analyzed benthic infauna, and conducted
oyster larval bioassays. With the exceptions of the NOAA neoplasm/disease
and EPA amphipod studies, MRL also statistically treated and interpreted
all data, and prepared this final report.
It should be understood that these results represent a reconnaissance
of the problems and are not always rigorously quantitative. Hypothesis
testing was not always possible and selected statistical tests were
sometimes exploratory and intended to detect patterns and trends in the
data. The strength of our strategy was in the performance of several
chemical analyses, biological analyses, and bioassays on the same fresh
(unfrozen) sediment sample. The development of a broad based (summary)
index of degraded sediment quality also facilitated detection of a wider
range of sediment toxicity that would not have been evident from applying
any single analysis or bioassay technique.
The major findings resulting from this report are summarized as
follows.
Bathymetry and Sediment Characteristics
The eight bays demonstrated different depth and sediment
characteristics. The Fourmile Rock - Elliott Bay dump site vicinity and
Dabob Bay were distinctly deeper (>100 m) and contained sediments that, on
the average, were sandier and of lower total organic carbon (TOC) content
than the rest. Samish and Bellingham Bays were the shallowest and the
muddiest of the eight bays. Bellingham Bay showed the second highest TOC
level. Case Inlet, Sinclair Inlet, and Sequim Bay were of intermediate
depth and had intermediate percentages of silt and clay and TOC. These
three bays exhibited sediment types ranging from sandy-silt to silty-mud.
Port Gardner - Everett Harbor was distinct from the other bays because it
had a significantly higher mean TOC level.
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Contaminant Levels
Urban bay sediments, particularly in Sinclair Inlet and at the
Fourmile Rock - Elliott Bay dump site vicinity, were found to be
contaminated by Ag, As, Cu, Cd, Hg, Pb, and Zn. Other metals (Sb, Be, Cr,
Ni, Se, Ti) were found at about the same concentration in all bays;
however, a few urban sediments in Bellingham Bay and Sinclair Inlet were
enriched in Sb, Cr, and Ni. Baseline bays generally contained lower
metals concentrations, many of which were found at approximately the same
concentrations as in pre-1900 sediments.
Urban bay sediments, particularly those from Sinclair Inlet and
Port Gardner - Everett Harbor, also contained significant quantities of
aromatic hydrocarbons and PCB-1254. Other organic compounds occasionally
found in urban bays included phthalates and PCB-1260. However, no PCBs
were detected in baseline bays, and only a few stations in Samish Bay
contained detectable quantities of phthalates and aromatic hydrocarbons.
Benthic Infauna
It was determined that in all sediment types, TOC levels between
4 and 16% were associated with a disturbed infauna dominated by "organic
enrichment opportunists." This shift to enrichment opportunists,
primarily nematodes and Capitella capitata, occurred at almost all
stations in Port Gardner - Everett Harbor and at several stations in
Bellingham Bay. The extent of this shift was less in sandy sediments when
compared to silty sediments.
After omitting the stations with high TOC, the numerically dominant
infauna were shown to vary more with sediment type than by bay, except
that Case Inlet and Samish Bay had distinctly different infaunas than all
other bays. Also, omitting these same stations did not reveal a
distinctly different set of numerically dominant infaunal species in urban
bays when compared with baseline bays.
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Fish and Shellfish Pathology
Flatfish species were not collected in four of the eight bays
(Bellingham Bay, Samish Bay, Dabob Bay, Sequim Bay) because of seasonal
scarcity, so the resulting pathology data were not used in a summary
statistical exercise to determine signs of degraded sediment quality.
However, these data served to corroborate the results of other approaches
used in the summary exercise (chemistry, infaunal analyses and amphipod
and oyster larval bioassays) indicating that urban bays were generally
more severely impacted than baseline bays.
English sole caught in Sinclair Inlet demonstrated the highest
incidence of liver, kidney, and gill lesions. Also, two types of lesions
in shellfish were found only in animals from urban bays. Hydropic
degeneration/membrane lysis in the hepatopancreas of Dungeness crab and
degenerative disorders in the antennal gland of two shrimp species were
observed in Bellingham Bay and at the Fourmile Rock - Elliott Bay dump
site vicinity. And, although the incidences of serious liver, kidney, and
gill lesions in English sole from Case Inlet were low compared to
incidences detected in sole from other urban bays (Duwamish Waterway),
these lesions were not detected in sole from Eliza Island (substituted
baseline station).
Amphipod Survival
Based on the amphipod survival data alone, the urban bays were not
always clearly distinguishable from baseline bays. The Fourmile Rock -
Elliott Bay dump site vicinity, representative of urban bay conditions,
and Sequim Bay, a baseline bay, demonstrated the highest and second
highest mean survivals of 17.3 and 17.0 respectively. Similarly, Case
Inlet, a baseline bay, and Port Gardner - Everett Harbor, an urban bay,
demonstrated the second lowest and lowest mean survivals of 13.0 and 12.3
respectively.
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Correlation analyses illustrated significant relationships between
the amphipod bioassay and various physical and chemical properties of
sediments. Of particular importance were the relationships of amphipod
survival to sediment grain size, percent water, organic content (percent
volatiles and TOC), and burden of organic compounds (PCB-1254, aromatic
hydrocarbons). All such properties were interpreted as accounting for
lowered amphipod survival. These analyses also revealed that lowered
amphipod survival correlated with increased abundance of organic
enrichment infaunal species in Port Gardner - Everett Harbor.
Oyster Larval Bioassay
Results of the oyster larval bioassay generally paralleled those of
amphipod bioassays. The analyses, when taken alone, indicated that urban
bays were not always different from baseline bays. Although a majority of
stations in Bellingham Bay and Sinclair Inlet, both urban bays, exhibited
significantly higher mean percentages of abnormal larvae when compared
with controls, the Fourmile Rock - Elliott Bay dump site vicinity, which
was also considered representative of urban bay conditions, exhibited no
differences when compared with controls. Similarly, although Sequim Bay,
Samish Bay, and Case Inlet (all baseline bays) exhibited no differences
among mean percentages of abnormal larvae when compared with controls,
Dabob Bay, (another baseline bay) exhibited higher mean percentages of
abnormal larvae at two of four stations.
Correlation analyses showed a significant relationship between
percent abnormal oyster larvae and amphipod survival. The trend to higher
percentages of abnormal oyster larvae become more evident as amphipod
survival decreased.
Summary Statistics
On the basis of the summary exercise which evaluated all data,
chemical and biological, urban bays were significantly more impacted than
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baseline bays. The greatest impacts were found in Port Gardner - Everett
Harbor, Sinclair Inlet, and Bellingham Bay. The Fourmile Rock - Elliott
Bay dump site vicinity appeared to be the least impacted of the
urban-industrialized sampling areas. Not surprisingly, Sequim Bay was the
least impacted of all bays. However, Case Inlet and Samish Bay, both
baseline bays, showed some indication of degraded sediment quality. All
sediments in both bays resulted in lower than control amphipod survival.
The amphipod and oyster larval bioassay data from Dabob Bay also suggest
some degree of impact.
It was also observed that the most adversely impacted stations in
Port Gardner - Everett Harbor were located in the East Waterway. For
Sinclair Inlet, they were located in proximity to the Puget Sound Naval
Shipyard. In Bellingham Bay, they were associated with the inner harbor.
Relationship of Urban to Baseline (Reference) Bay Concept
On the basis of chemical contaminants alone, the baseline or
"reference bay" concept appears to be valid. Sequim Bay or Dabob Bay,
with relatively low metals burdens and no detectable organic compounds,
seem to be excellent choices for reference bays.
Biological variables, however, do not always clearly distinguish
between what we hypothesized at the outset of the study to be urban bays
and baseline bays. Rather, the biological findings better describe a
continuum of responses consistent with a gradient of contaminants existing
between regions of higher and lower contamination in Puget Sound.
This continuum is described here: 1) Port Gardner - Everett Harbor,
Bellingham Bay, and Sinclair Inlet occupy one end of the continuum,
representing water bodies now showing the most evidence of degraded
sediment quality. 2) Sequim Bay is a baseline bay located at the opposite
end of the continuum. It presently shows the least indication of degraded
sediment quality of all the bays sampled. 3) The Founnile Rock - Elliott
Bay dump site vicinity is located intermediate on the continuum but closer
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to the position occupied by the urban bays. However, clearly the dump
site is somewhat anomalous because not one of the eight stations sampled
during the detailed surveys showed any sign of degraded sediment quality
when evaluated by either the amphipod or oyster larval bioassay. 4) Case
Inlet and Samish Bay are also located intermediate on the continuum but
closer to the baseline bay position occupied by Sequim Bay. However, both
bays are close enough to Commencement Bay and Bellingham Bay respectively
to show some signs of degraded sediments. 5) Dabob Bay is also located
intermediate on the continuum but closest to the baseline bay position
occupied by Sequim Bay. The finding of degraded sediment quality based
again on the bioassay results is perhaps attributed to circulation of
contaminants from the main basin of Puget Sound.
In summary, the results of our study support the "reference bay"
concept; however, our results also suggest that the process to select
reference bays needs refinement. It is evident that selection should be
based on chemical, biological, and physical data. Also, the 1983
screening survey was particulary useful in providing relevant and recent
data on which to base the selection process. Ideally, a reference bay
should contain little or no chemical contamination, demonstrate few or no
impacts when evaluated by infaunal analyses and sediment bioassays, but
should also possess physical characteristics (depth and grain size) that
closely approximate the urban bay(s) to be examined. Our results perhaps
suggest that "reference stations" may be a more appropriate concept than
"reference bays", particularly when time and funds limit the scope of
studies performed in support of regulatory action.
There is a need to bring together a forum of scientists and
regulators to refine this concept and its intended use. Also, our strategy
of selecting equal numbers of the most contaminated stations within each
urban bay does not address issues concerning the full range of
contaminated sediments within each bay. Clearly, the present study would
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have benefitted from the addition of stations both in the urban and
baseline bays. Consequently, our results apply only to the sediments
(stations) sampled in each bay and do not likely represent all similiar
depositional sediments in each bay.
Usefulness of the Reconnaissance Survey Concept
The 1983 screening survey of 181 stations from eight bays was
particularly useful in appropriately focusing the 1984 detailed survey.
The strategy of selecting the most contaminated stations from the urban
bays and the least contaminated stations from the baseline bays permitted
limited resources to be directed toward identifying those stations most
likely to reveal degraded sediment quality. Moreover, the screening
survey data allowed the station selection process for the detailed surveys
to be based on data that was recently acquired rather than on historical
data that was collected in support of other goals. The variability
encountered in biological indices subsequently revealed the wisdom of the
selection strategy applied. Another selection strategy would have likely
facilitated less clear-cut biological findings. Consequently, we have
learned that sampling strategies must be developed specifically for each
study; and if faced with another large sediment survey, we would again
recommend that a preliminary or reconnaissance level effort be undertaken
to determine which sampling strategy best addressed the study goals within
available resources.
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CONTENTS
ACKNOWLEDGEMENTS iii
EXECUTIVE SUMMARY vii
1.0 INTRODUCTION ..... 1
1.1 OBJECTIVES AND SCOPE 3
2.0 METHODS AND MATERIALS 7
2.1 FIELD COLLECTION PROCEDURES 7
2.1.1 Screening Surveys 7
2.1.2 Detailed Surveys 21
2.2 PHYSICAL AND CHEMICAL (RANGE FINDING) SEDIMENT
ANALYSES (PERFORMED BY MRL) 23
2.2.1 Grain Size 23
2.2.2 Total Extractable Hydrocarbons Determined
by Infrared Spectrophotometry 24
2.2.3 Water Content 24
2.2.4 Volatile Solids 25
2.2.5 Total Organic Carbon 25
2.2.6 Metals Analyses 26
2.3 PRIORITY POLLUTANT SEDIMENT ANALYSES (PERFORMED
BY ErAJ ........... Cy
2.3.1 Procedures for Organic Compounds . . . . .29
2.3.2 Procedures for Metals ....... 32
2.3.3 Procedures for Solids (Percent) 33
2.3.4 Procedures for Phenolics and Cyanides . . . .33
2.3.5 Detection and Quantitation Limits . . . .33
2.3.6 Precision, Accuracy, and Completeness . . . .34
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2.4 BENTHIC INFAUNAL ANALYSES (PERFORMED BY MRL) ... 44
2.4.1 General Procedures ....... 44
2.4.2 Statistical Treatment 45
2.5 FISH AND INVERTEBRATE PATHOLOGY (PERFORMED BY NOAA) . . 45
2.5.1 Fish Capture and Necropsy 45
2.5.2 Fish Histologic and Diagnostic
Procedures 47
2.5.3 Crab and Shrimp Capture and Necropsy . . . .47
2.5.4 Crab and Shrimp Histological and
Diagnostic Procedures 48
2.5.5 Statistical Treatment 48
2.6 AMPHIPOD BIOASSAY (PERFORMED BY EPA) 48
2.6.1 Strategy ......... 48
2.6.2 Test Organisms and Control Sediments . . . .49
2.6.3 Screening Surveys ........ 49
2.6.4 Detailed Surveys 50
2.6.5 Statistical Treatment 51
2.7 OYSTER LARVAL BIOASSAY (PERFORMED BY MRL) 52
2.7.1 Strategy 52
2.7.2 Screening Surveys 52
2.7.3 Detailed Surveys ........ 53
2.7.4 Statistical Treatment 54
2.8 GENERAL STATISTICAL PROCEDURES (PERFORMED BY MRL) ... 54
2.8.1 Discriminant Analysis 54
2.8.2 Factor Analysis 55
2.8.3 Correlation Analysis 56
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2.8.4 Tolerance Intervals 57
2.8.5 Summary Statistics 57
3.0 RESULTS 61
3.1 SEDIMENTARY CHARACTERISTICS (PERFORMED BY MRL) ... 61
3.1.1 Screening Surveys 61
3.1.2 Detailed Surveys 64
3.2 CONTAMINANT LEVELS (PERFORMED BY BOTH EPA
AND MRL) 67
3.2.1 Screening Surveys 67
3.2.2 Detailed Surveys 69
3.2.3 Statistical Treatment 79
3.3 BENTHIC INFAUNA (PERFORMED BY MRL) 84
3.3.1 Bathymetry and Sediment Types 84
3.3.2 Richness and Abundance 84
3.3.3 General Infaunal Patterns 90
3.3.4 Infaunal Patterns in Each Bay . . . . .93
3.3.5 Infaunal Characteristics by Sediment Type . . . 101
3.3.6 Statistical Treatment ....... 109
3.4 FISH AND SHELLFISH PATHOLOGY (PERFORMED BY NOAA) . . .110
3.4.1 Fish Pathology 110
3.4.2 Shellfish Pathology 121
3.5 AMPHIPOD BIOASSAY (PERFORMED BY EPA) 134
3.5.1 Screening Surveys 134
3.5.2 Detailed Surveys 146
3.6 OYSTER LARVAL BIOASSAY (PERFORMED BY MRL) . . . . 167
3.6.1 Screening Surveys ........ 167
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3.6.2 Detailed Surveys 167
3.7 SUMMARY STATISTICS 130
4.0 DISCUSSION .185
4.1 CONTAMINANT LEVELS 185
4.1.1 Relationship to Physical Properties of
Sediments 185
4.1.2 Contaminants of Urban vs. Baseline Bays . . .186
4.1.3 Comparisons to Other Studies 186
4.2 BENTHIC INFAUNA 190
4.2.1 Relationship to Physical and Chemical
Properties of Sediments ...... 190
4.2.2 Infaunal Patterns of Urban and
Baseline Bays 190
4.3 FISH AND SHELLFISH PATHOLOGY 194
4.3.1 Relationships Between Lesion Incidence
and Sediment Chemical Composition .... 194
4.3.2 Comparison of Fish and Shellfish Pathology
in Urban and Baseline Bays 199
4.3.3 Comparisons with Other Studies 201
4.4 AMPHIPOD BIOASSAY 204
4.4.1 Relationship to Physical and Chemical
Properties of Sediments 204
4.4.2 Comparison of Amphipod Survival in Urban
and Baseline Bays 205
4.4.3 Comparisons to Other Studies 207
4.5 OYSTER LARVAL BIOASSAY 208
4.5.1 Relationship to Physical and Chemical
Properties of Sediments ...... 208
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4.5.2 Comparison of Oyster Larval Abnormalities
in Urban and Baseline Bays 209
4.5.3 Comparison to Other Studies. 209
4.6 SUMMARY STATISTICS " . . .210
4.7 RELATIONSHIP OR URBAN TO BASELINE (REFERENCE) BAY
CONCEPT 212
5.0 CONCLUSIONS 215
5.1 BATHYMETRY AND SEDIMENT CHARACTERISTICS 215
5.2 CONTAMINANT LEVELS 215
5.3 BENTHIC INFAUNA 216
5.4 FISH AND SHELLFISH PATHOLOGY 216
5.5 AMPHIPOD SURVIVAL 217
5.6 OYSTER LARVAL BIOASSAY 217
5.7 SUMMARY STATISTICS 218
5.8 RELATIONSHIP OF URBAN TO BASELINE (REFERENCE)
BAY CONCEPT 219
5.9 USEFULNESS OF THE RECONNAISSANCE SURVEY CONCEPT . . .221
6.0 REFERENCES 223
APPENDIX A - STATION IDENTIFICATION AND PROTOCOL FOR
STATION SELECTION A.I
APPENDIX B - SEDIMENTARY CHARACTERISTICS AND CONTAMINANT
LEVELS B.I
APPENDIX C - AMPHIPOD BIOASSAY C.I
APPENDIX D - OYSTER LARVAL BIOASSAY D.I
APPENDIX E - DATA MATRIX E.I
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FIGURES
1 Puget Sound ........... 2
2 Bellingham Bay Sampling Stations (Inner Harbor) .... 8
3 Bellingham Bay Sampling Stations (Outer Harbor) .... 9
4 Port Gardner - Everett Harbor Sampling Stations . . . .10
5 Sinclair Inlet Sampling Stations 11
6 Fourmile Rock - Elliott Bay Dump Site Vicinity
Sampling Stations 12
7 Samish Bay Sampling Stations 13
8 Case Inlet Sampling Stations 14
9 Dabob Bay Sampling Stations 15
10 Sequim Bay Sampling Stations 16
11 Percent of Silt and Clay in Sediments 65
12 Percent of Volatiles in Sediments 65
13 Percent of TOC in Sediments 66
14 IR Results for Organics in Sediments 66
15 Silver Concentrations in Sediments 71
16 Arsenic Concentrations in Sediments 71
17 Copper Concentrations in Sediments 72
18 Mercury Concentrations in Sediments 72
19 Lead Concentrations in Sediments 73
20 Zinc Concentrations in Sediments 73
21 Cadmium Concentrations in Sediments 74
22 Chromium Concentrations in Sediments 74
23 Nickel Concentrations in Sediments 75
24 Aromatic Hydrocarbons Concentrations in Sediments . . .75
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25 PCB-1254 Concentrations in Sediments 76
26 Discriminant Analysis of 8 Bays Using the Canonical
Variables Ag, IR, and As 80
27 Discriminant Analysis of 48 Stations from 8 Bays
Using the Canonical Variables Ag, IR, and As .... 81
28 Mean Depth Versus Mean Percent Silt and Clay by Bay
(48 Stations) 86
29 Mean TOC Versus Mean Percent Silt and Clay (48
Stations) 87
30 Differences Among Bays According to Depth, Sediment
Type and TOC (48 Stations) 88
31 Mean Number of Individuals per Station Versus Mean
Number of Taxa per Station (48 Stations) 91
32 Results of Amphipod Bioassays on Sediments Collected
in Bellingham Bay During Screening Surveys (August 2
to September 18, 1983) 135
33 Results of Amphipod Bioassays on Sediment Collected
in Port Gardner - Everett Harbor During Screening
Surveys (August 2 to September 18, 1983) 136
34 Results of Amphipod Bioassays on Sediments Collected
in the Fourmile Rock - Elliott Bay Dump Site Vicinity
During Screening Surveys (August 2 to September 18,
1983) ............ 137
35 Results of Amphipod Bioassays on Sediments Collected
in Sinclair Inlet During Screening Surveys (August 2
to September 18, 1983) 138
36 Results of Amphipod Bioassays on Sediments Collected
in Samish Bay During Screening Surveys (August 2 to
September 18, 1983) 139
37 Results of Amphipod Bioassays on Sediments Collected
in Case Inlet During Screening Surveys (August 2 to
September 18, 1983) 140
38 Results of Amphipod Bioassays on Sediments Collected
in Dabob Bay During Screening Surveys (August 2 to
September 18, 1983) 141
• xxi
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39 Results of Amphipod Bioassays on Sediments Collected
in Sequim Bay During Screening Surveys (August 2 to
September 18, 1983) 142
40 Results of Amphipod^Bioassays on Sediments Collected
in Bellingham Bay During Detailed Surveys (April 23
to May 29, 1984) 147
41 Results of Amphipod Bioassays on Sediments Collected
in Port Gardner - Everett Harbor During Detailed
Surveys (April 23 to May 29, 1984) 148
42 Results of Amphipod Bioassays on Sediments Collected
in the Fourmile Rock - Elliott Bay Dump Site Vicinity
During Detailed Surveys (April 23 to May 29, 1984) . . .149
43 Results of Amphipod Bioassays on Sediments Collected
in Sinclair Inlet During Detailed Surveys (April 23
to May 29, 1984) 150
44 Results of Amphipod Bioassays on Sediments Collected
in Samish Bay During Detailed Surveys (April 23 to
May 29, 1984) 151
45 Results of Amphipod Bioassays on Sediments Collected
in Dabob Bay During Detailed Surveys (April 23 to
May 29, 1984) 152
46 Results of Amphipod Bioassays on Sediments Collected
in Case Inlet During Detailed Surveys (April 23 to
May 29, 1984) 153
47 Results of Amphipod Bioassays of Sediments Collected
in Sequim Bay During Detailed Surveys (April 23 to
May 29, 1984) 154
xxii
-------
TABLES
1 Locations of Stations at Which Target Fish Species Were
Captured and Necropsied, March 1984 17
2 Locations of Stations at Which Target Crab and Shrimp
Species were Captured and Necropsied, March, 1984 . . .18
3 Stations Selected for Detailed Surveys . . . . .21
4 Detection Limits, Blanks, and Sediment Standards
Analyzed by MRL (ppm dry weight) 28
5 Average Quantitation Limits of Organic Compounds
Analyzed in Puget Sound Sediment Samples . . . . .35
6 Detection Limits of Metals in Sediments
(1.0 g/100 ml) 37
7 Quality Control Samples Analyzed 37
8 Average Percent Recovery of Surrogate Compounds of
VOA Fraction 38
9 Percent Recovery of VOA Spikes 39
10 Precision of VOA Spikes (RPD) 39
11 Average Recovery Acid/Base-Neutral Surrogate
Standards 40
12 Matrix Spike Percent Recovery of the Acid/
Base-Neutral Fraction 42
13 Precision of Acid/Base-Neutral Spikes (RPD) 43
14 Mean Sediment Characteristics and Contaminant
Levels for 48 Stations Sampled During Screening
(1983) and Detailed Surveys (1984) Determined
by MRL 62
15 Coefficients for Canonical Variables Resulting from
Discriminant Analyses Performed on Physical and
Chemical Characteristics of Sediments Collected at
48 Stations 79
16 Sorted Rotated Factor Loadings for 13 Sediment
Characteristics and Contaminants for 48 Stations . . . .83
xxm
-------
17 Summary of the Distribution of Sediment Types,
Depth, and Other Characteristics among the Eight
Bays (48 Stations) 85
18 Mean Values for Number of Taxa and Number of
Individuals per Station from 48 Stations
within 8 Puget Sound Embayments 89
19 The Ten Most Abundant Taxa for All Bays 92
20 Comparison of the Bays by Dominant Benthic Infauna . . .94
21 Comparison of the Dominant Benthic Infauna by
Sediment Type 101
22 Comparison of the Dominant Benthic Infauna by
Sediment Type (Stations with High TOC and
Stations from Case Inlet and Samish Bay Omitted) .... 106
23 Incidences (%) of Idiopathic Liver Lesions in
English Sole from Stations Sampled in Puget
Sound, March 1984 Ill
24 Incidences (%) of Idiopathic Liver Lesions in
English Sole from Reference and Urban Areas by
Size Class 113
25 Incidences (%) of Idiopathic Kidney and Gill
Lesions in English Sole 116
26 Incidences (%) of Idiopathic Liver Lesions in
Dover Sole from Elliott Bay 120
27 Incidences (%) of Idiopathic Kidney and Gill
Lesions in Dover Sole from Elliott Bay 121
28 Incidences (%) of Individual Histopathological
Conditions in Cancer magister 124
29 Incidences (%) of Major Histopathological
Categories in Cancer magister, Bellingham Bay,
Samish Bay, and Dungeness Spit 126
30 Incidences (%) of Individual Histopathological
Conditions in Cancer gracilis 127
31 Incidences (%) of Major Histopathological
Categories in Cancer gracilis from Case and
and Sinclair Inlets ......... 129
xxiv
-------
32 Incidences (%) of Individual Histopathological
Conditions in Shrimp from Elliott and Dabob Bays : 130
33 Incidences (%) of Major Histopathological Categories
for Shrimp from Elliott and Dabob Bays 132
34 Lesion and Tissue Codes used in Tables 28-33 . . . .133
35 Mean Values (± Standard Deviation) for 1983
Amphipod Survival and Sediment Characteristics
for All Bays (180 Stations) 143
36 Correlation Matrix for Numbers of Amphipods
Surviving (1983) and Sediment Characteristics
from All Bays (180 Stations) 144
37 Results of Regression Analyses for Numbers
of Amphipods Surviving (1983) and Sediment
Characteristics from all Bays (180 Stations) . . . .144
38 Mean Percent H20 and Percent Fines at Various
Survivor Levels 145
39 Summary of Statistical Analyses (HOMOV, ANOVA,
SNK, Dunnett's Test) of Survival and Reburial Data
for Rhepoxynius abronius Exposed to Puget Sound
Sediments (Assay No. 1 - Bellingham Bay [B-l],
Samish Bay [B-2]) 155
40 Summary of Statistical Analyses (HOMOV, ANOVA,
SNK, Dunnett's Test) of Survival and Reburial Data
for Rhepoxynius abronius Exposed to Puget Sound
Sediments (Assay No. 2 - Bellingham Bay [B-l],
Port Gardner - Everett Harbor [B-4]) 156
41 Summary of Statistical Analyses (HOMOV, ANOVA,
SNK, Dunnett's Test) of Survival and Reburial Data
for Rhepoxynius abronius Exposed to Puget Sound
Sediments (Assay No. 3 - Port Gardner - Everett
Harbor [B-4], Fourmile Rock - Elliott Bay Dump
Site Vicinity [B-5]) 157
42 Summary of Statistical Analyses (HOMOV, ANOVA,
SNK, Dunnett's Test) of Survival and Reburial Data
for Rhepoxynius abronius Exposed to Puget Sound
Sediments (Assay No. 4 - Fourmile Rock - Elliott
Bay Dump Site Vicinity [B-5], Sinclair Inlet [B-6]) . . .158
xxv
-------
43 Summary of Statistical Analyses (HOMOV, ANOVA,
SNK, Dunnett's Test) of Survival and Reburial Data
for Rhepoxynius abronius Exposed to Puget Sound
Sediments (Assay No. 5 - Sinclair Inlet [B-6], Dabob
Bay [B-7]) 159
44 Summary of Statistical Analyses (HOMOV, ANOVA,
SNK, Dunnett's Test) of Survival and Reburial Data
for Rhepoxynius abronius Exposed to Puget Sound
Sediments (Assay No. 6 - Sequim Bay [B-3], Case
Inlet [B-8]) 160
45 Significance of Amphipod Survival Data by Bay
and Station for Sediments Collected During
Detailed Surveys April 23 to May 29, 1984 161
46 Chi-Square 2x2 Analysis to Test for Batch Effect
in Amphipod Test Populations 163
47 Mean Values (± Standard Deviation) for 1984 Amphipod
Survival and Sediment Characteristics for All Bays
(48 Stations) 164
48 Comparison of Procedure with Random Chance, All
48 Stations 166
49 Comparison of Procedure with Random Chance,
13 Stations with Inconclusive Results (Mean
Survival Between 15.1 and 16.9) Removed 166
50 Effects of Sediment Concentration on Oyster Larval
Development as Percent Abnormals (Assays Performed
August 24 to September 24, 1983 on Fresh Puget Sound
Sediments) 168
51 Effects of Puget Sound Sediments on Oyster Larval
Development Measured as Percent Abnormals (Assay
No. 1 Performed April 26 to 28, 1984) 169
52 Effects of Puget Sound Sediments on Oyster Larval
Development Measured as Percent Abnormals (Assay
No. 2 Performed May 10 to 12, 1984) 170
53 Effects of Puget Sound Sediments on Oyster Larval
Development Measured as Percent Abnormals (Assay
No. 3 Performed May 10 to 12, 1984) 171
54 Effects of Puget Sound Sediments on Oyster Larval
Development Measured as Percent Abnormals (Assay
No. 4 Performed May 17 to 19, 1984) 172
xxvi
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55 Effects of Puget Sound Sediments on Oyster Larval
Development Measured as Percent Abnormals (Assay
No. 5 Performed May 24 to 26, 1984) 173
56 Effects of Puget Sound Sediments on Oyster Larval
Development Measured as Percent Abnormals (Assay
No. 6 Performed June 4 to 6, 1984) 174
57 Summary Analysis of Variance for Percent Abnormality
(Arcsin /X) in Each Oyster Larval Bioassay 175
58 Comparison of Treatment and Control Abnormality
(Arcsin /X) Using Dunnett's Test 177
59 Significance of Oyster Larval Percent Abnormality
Data by Bay and Station for Sediments Collected
During Detailed Surveys April 23 to May 29, 1984 . . . .178
60 Summary Scores for Chemical, Biological, and Physical
Variables Used to Determine Signs of Degraded
Sediment Quality 182
61 Comparisons of Concentrations of Contaminants in
Puget Sound Sediments 187
62 Comparison of Concentrations of Metals in Surface
Sediments from Selected Areas of Puget Sound (ppm
dry weight) 189
xxvi i
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RECONNAISSANCE LEVEL ASSESSMENT
OF SELECTED SEDIMENTS FROM PUGET SOUND
1.0 INTRODUCTION
Puget Sound is one of the most biologically productive and
recreationally important fjord-like estuaries in the continental United
States. Although most of Puget Sound is considered clean, healthy, and
productive, serious water quality problems exist in certain areas,
particularly in the more urbanized embayments. In addition, as the region
continues to grow and develop, the potential cumulative effects of waste
discharges on the environmental quality of Puget Sound as a whole become a
concern. This type of cumulative effect markedly reduced the productivity
of Chesapeake Bay during the 1960s. Major economic uses of Puget Sound,
such as commercial and sport fisheries, tourism and recreation, depend on
maintaining a high level of water quality. Accordingly, existing and
potential water quality degradation represent a serious threat to these
important uses of Puget Sound.
The eastern shore of Puget Sound from Tacoma to Everett is the most
densely populated area of the Pacific Northwest. The waters of Puget
Sound (see Figure 1) receive the wastes of approximately 2.5 million
people, as well as wastes from the outfalls of numerous industrial
facilities associated with pulp and paper, other forest products, food
processing, metals and petroleum refining, and chemical manufacturing. In
addition to these point sources of pollution, nonpoint sources of
pollution diffuse into Puget Sound from groundwater, surface runoff from
adjacent land areas, return flow from irrigated fields, and atmospheric
fallout. These many sources of pollution to Puget Sound combine to alter
water and sediment quality (Pavlou and Dexter 1979; Riley et al. 1980,
1981), impact or potentially impact biological communities (Bingham 1978;
Swartz et al. 1982), and limit beneficial uses of surface waters (Maiins
-------
BELLINGHAM
Bellingham Bay
Samish Bay
Everett Harbor
Port Gardner
PORT Seguim Bay
ANGELES
Elliott Bay
SHELTON
TACOMA
OLYMPIA
FIGURE 1. Puget Sound.
-------
et al. 1980, 1982). These sources of pollution and concern for vulnerable
resources emphasize the need to reevaluate the procedures in place for
managing Puget Sound water quality.
The U.S. Environmental Protection Agency (EPA) and the Washington
Department of Ecology (WDOE), as primary environmental regulatory agencies
in the Puget Sound area, recognized this need and, in 1983, designed and
implemented a program to focus attention and interagency resources on
solving the most pressing Puget Sound water quality problems.
With the consent of WDOE, EPA contracted with Pacific Northwest
Laboratory (PNL) on July 9, 1983, to conduct a study to better define the
significance of pollution in certain urban-industrial bays in Puget Sound
(Bellingham Bay, Port Gardner - Everett Harbor, Sinclair Inlet, and the
Fourmile Rock - Elliott Bay dump site vicinity). The EPA and WDOE
envisioned that this could best be achieved by first defining reference
levels of contamination in several relatively undeveloped or baseline
embayments in Puget Sound (Samish Bay, Case Inlet, Dabob Bay, Sequim Bay).
At the same time, EPA and WDOE asked PNL to estimate the mass loading of
contaminants to Port Gardner - Everett Harbor and to determine the rate of
accumulation of contaminants in the local sediments so as to direct
possible corrective actions (report submitted). Also, as part of this
contract, PNL studied existing institutional arrangements and procedures
for water quality management of Puget Sound (Morris et al. 1984).
1.1 OBJECTIVES AND SCOPE
The objective of this effort was to develop a better understanding of
the toxic contamination problems in selected urban-industrialized bays of
Puget Sound. Understanding these problems will aid in developing a basis
for additional regulatory action.
Two major tasks comprised this effort. The first was to conduct
preliminary or screening surveys and analyze sediment samples collected
from 101 stations distributed in four urban-industrialized embayments
(Bellingham Bay, Port Gardner - Everett Harbor, Fourmile Rock - Elliott
-------
Bay dump site vicinity, and Sinclair Inlet) and at 80 stations distributed
in four baseline embayments (Samish Bay, Case Inlet, Dabob Bay, Sequim
Bay). The screening surveys were used as a guide for more detailed
technical analyses. The second task involved detailed surveys and
analyses of the same bays, but at a limited number of stations (32 in
urban embayments, 16 in baseline bays). The 1984 sediment samples
selected for detailed analyses were the "cleanest" of the clean and the
"dirtiest" of the dirty as determined by the 1983 sediment chemical
analyses, and within restrictions imposed by sediment type.
It should be understood that these studies represent a reconnaissance
of the problem and are not always rigorously quantitative. Hypothesis
testing was not always possible and selected statistical tests were
exploratory and designed to detect patterns and trends in the data. The
strength of our strategy was the conduct of several chemical analyses,
biological analyses, and bioassays on the same fresh (unfrozen) sediment
sample.
Based on the results of these surveys, this final report describes
the sediment chemistry, sediment toxicity, numerically dominant benthic
infauna, and incidence of fish and shellfish disease in each urban and
baseline bay. Each biological parameter (benthic infauna, incidence of
fish and shellfish disease, amphipod and oyster larval bioassay) is
discussed in relation to the physical and chemical properties of
associated sediments. Present research findings are also compared with
those of historical surveys. Finally, the embayments and the stations in
each embayment that currently show signs of degraded sediment quality are
identified.
The EPA Region 10 Laboratory, the Northwest and Alaska Fisheries
Center of NOAA, and the Marine Research Laboratory (MRL) of PNL
participated in developing material included in this report. The EPA
Region 10 Laboratory staff helped conduct screening surveys in the fall of
1983 and conducted the detailed surveys in the spring of 1984. They also
performed priority pollutant analyses and amphipod bioassays. The NOAA
Northwest and Alaska Fisheries Center conducted independent surveys in the
same urban and baseline embayments to obtain fish and shellfish for
-------
analyses of neoplasia and other diseases. The MRL helped conduct the
screening surveys, performed range-finding chemistry for both the
screening and detailed surveys, analyzed benthic infauna, and conducted
oyster larval bioassays. With the exceptions of the NOAA neoplasia/
disease and EPA amphipod studies, MRL also statistically treated and
interpreted the data, and prepared the final report.
-------
2.0 METHODS AND MATERIALS
2.1 FIELD COLLECTION PROCEDURES
This section describes the procedures used to conduct the screening
and detailed surveys.
2.1.1 Screening Surveys
2.1.1.1 Sampling Areas and Locations
The locations of most sampling stations in urban bays were chosen on
the basis of their proximity to point sources of pollutants, or on the
basis of previously performed chemical analyses of bottom sediments from
these areas. Other locations in urban bays were chosen on the basis of
sediment type; that is, depositional sediments with a high mud to sand
ratio were selected for sampling. Such depositional sediments often act
as "sinks" for industrial chemicals (Wildish et al. 1980). Sampling
stations in baseline bays were selected solely on the basis of sediment
type. Again, sediments with a high mud to sand ratio were selected for
sampling.
The locations of 101 sampling stations in the urban-industrialized
bays of Puget Sound (Bellingham Bay, Port Gardner - Everett Harbor,
Four-mile Rock - Elliott Bay dump site vicinity, and Sinclair Inlet) are
shown in Figures 2-6. Figures 7-10 show the locations of the 80 sampling
stations in the baseline bays (Samish Bay, Case Inlet, Dabob Bay, and
Sequim Bay). Detailed descriptions (latitude, longitude, and depth) of
each sampling station are given in Appendix A. Other stations sampled, in
both urban and baseline bays, to determine incidence of fish and shellfish
disease are described in Tables 1 and 2. An adequate sample size of fish
or shellfish could not always be obtained at preselected sampling
stations.
-------
BELLINGHAM BAY
Samplings
01983
• 1983,1984
FIGURE 2. Bellingham Bay Sampling Stations (Inner Harbor)
8
-------
BELLINGHAM BAY
Samplings
91983
• 1983,1984
FIGURE 3. Bellingham Bay Sampling Stations (Outer Harbor)
-------
EVERETT HARBOR
/ Samplings
O1983
• 1983,1984
FIGURE 4. Port Gardner - Everett Harbor Sampling Stations
10
-------
Samplings
O1983
•1983,1984
FIGURE 5. Sinclair Inlet Sampling Stations
11
-------
47°-38'N
o7
4
O
V2
Nautical Miles
Magnolia Bluff
47°-36'N
o2 o1
•9
12
i uo* Disposal Area
• *—'' *24
10 «17
16
O
20
23
»22
m
91 9O
I
Samplings
O1983
•1983,1984
013
ELLIOTT BAY
122°-26'W
FIGURE 6. Fourmile Rock - Elliott Bay Dump Site
Vicinity Sampling Stations
12
-------
°is SAMISH BAY
Samplings
O1983
• 1983,1984
FIGURE 7. Samish Bay Sampling Stations
13
-------
HARTSTENE ISLAND
Samplings
01983
• 1983,1984
FIGURE 8. Case Inlet Sampling Stations
14
-------
Samplings
O1983
• 1983,1984
FIGURE 9. Dabob Bay Sampling Stations
15
-------
SEQUIMBAY °
Samplings
O1983
• 1983,1984
FIGURE 10. Sequim Bay Sampling Stations
16
-------
TABLE 1. Locations of Stations at Which Target Fish Species Were Captured
and Necropsied, March 1984
Area
Case Inlet
Bellingham-
Samish Bays
(Eliza Island)
Dabob Bay
Sinclair Inlet
Elliott Bay3
Location
7°17'20"N X 122°50'3't"W
48017'20"N X 122°50'3'tllW
47°46'21.0"N X 122°51'12.6"W
47°32'54.9"N X 122°39'56.9"W
47°33'27.1"N X 122°38'03.2"W
47°33'25.5"N X 122°37'50.4"W
47033'25.5"N X 122°25M2.1"W
47°37'10.8"N X 122°24'08.9"W
47040'00"N X 122°25'55"W
47°35'38"W X 122°23'51"W
ID #
Case 5
NA2
Dabob 15
Sinclair 6
Sinclair 17
Sinclair 18
Elliott 17
Elliott 23
NA2
NODC1
Station *
12134
4015
7026
8010
8009
8006
10061
10063
10065 (West Pt.)
10064 (Duwamish Head)
Total Fish Examined
English Sole
(Parophrys
vetulus)
30
28
1
30
7
23
5
12
2
138
Dover Sole
(Microstomus
pacificus)
—
_._
—
30
20
30
3
83
1 National Ocean Data Center.
2 NA - Not applicable.
3 Dump site vicinity.
-------
CO
23
TABLE 2. Locations of Stations at which Target Crab and Shrimp Species were
Cancer Cancer Pandalopsis Pandalus
dlsparplatyceros
30
30
17
30
30
Captured and Necropsied, March 1984
Depth NOOC1
Area
Case Inlet
Bellingham
Bay
Samish Bay
Sinclair
Inlet
Oungeness
Spit
Elliott Bay2
Oabob Bay
Location
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2.1.1.2 Logistics and General Sampling Procedures
All 181 of the screening sediment samples were collected during an
extended cruise over the period from August 2, 1983, through September 15,
1983. The research vessel, Cathlamet Bay, was leased by Battelle and
manned by MRL and EPA staff. The EPA was responsible for operating the
vessel and documenting sampling location. The MRL staff was responsible
for sample acquisition and documentation.
Each sampling site was located using a precision navigational system
®
(Mini-Ranger ), and was represented by a single composite sample collected
and prepared on the research vessel. The samples were collected through
multiple casts (2-5) with a modified 0.1-m2 van Veen grab sampler to
acquire a 26-L composite sample suitable for analysis. The following
sampling procedure was used. The material from each cast was immediately
examined visually while in the sampler. Samples not achieving a minimium
penetration depth of 10 cm, containing boulders, or excessive amounts of
gravel or foreign objects (tires, wood, metal) were rejected and the cast
retaken. Acceptable casts were emptied into a sample compositing chamber
(a schedule-316, stainless steel container with orifices for decanting).
After the appropriate volume was collected and allowed to stand for 5 min,
seawater was decanted from the sediment. The sediment was then mixed
manually with a contaminant-free stainless steel paddle. The assumption
that the sample was well mixed was validated by replicate analysis for
selected subsamples, as follows: (1) three replicate subsamples for one
sample from each bay for analysis for carbon tetrachloride (CCK)
extractable organics, sediment grain size and percent volatiles; (2) three
replicate subsamples for one sample from each urban bay for metals
analysis.
To prevent contamination between samples, the van Veen grab sampler,
the composite sample chamber, and mixing paddle were cleaned thoroughly
after collection of each 26-L sample. The samples were maintained in the
dark and on ice at a temperature of 4°C. As an additional quality
control measure paper wipes were collected at the beginning of each day to
Registered trademark of the Motorola Company.
19
-------
ensure the cleanliness of the composite sampling chamber and sampling
containers, respectively. Control of other potential sources of variation
are addressed in the specific analytical protocols.
The subsample aliquots were placed into appropriately labeled
containers on board the research vessel as a part of the sampling
protocol. The containers were marked with a serial number representing a
bay code number (1 through 8), a station code number (1 through 26) and a
sample replicate code number (e.g., 1-2-3 where 1 = Bellingham Bay;
2 = Station No. 2 coordinates, 48° 45' 28.2"N, 122° 30' 47.0"W; and
3 = replicate number). During the course of the cruise day, the
appropriate size aliquots were taken from the samples and the containers
holding aliquots were sealed.
The EPA prepared and provided the appropriate containers for priority
pollutant analyses and amphipod bioassay. All other containers were
provided and prepared by the MRL.
2.1.1.3 Sample Custody
During the cruise day the samples were in custody of the MRL Senior
Scientist on board the vessel. At the close of the cruise day, collection
data were inscribed on each aliquot container, the subsamples were signed
by the Collector, and a list of samples and subsamples taken were logged
in MRL official laboratory record book(s) assigned to the project. The
laboratory record book also provided a record of the disposition of the
subsample. At this point, custody of subsamples for priority pollutant
analysis and amphipod bioassay was given to EPA. The remaining subsamples
were delivered to the MRL in Sequim, Washington. There, they were given
to the appropriate Task Leaders who logged the receipt of these samples
into the appropriate laboratory record books. As analyses were completed,
the data pertaining to those samples were also logged into the Task
Leaders' laboratory record books.
20
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2.1.2 Detailed Surveys
2.1.2.1 Sampling Areas and Locations
Table 3 presents the 48 stations that were revisited and resampled
for the detailed surveys. A protocol for choosing stations was developed
that selected the most contaminated stations from the urban bays and the
least contaminated stations from the baseline bays, based on the chemical
data derived from the screening surveys. The procedures and results of
applying this protocol to physical and chemical data generated during the
screening surveys are presented in Appendix A.
TABLE 3. Stations Selected for Detailed Surveys
Station Numbers
Bel lingham
3
4
5
7
11
12
23
24
Elliott
9
10
12
17
20
22
23
24
Everett
1
2
3
4
5
6
7
11
Sinclair
6
7
8
14
17
18
19
20
Samish
1
3
7
20
Da bob
1
5
7
15
Sequim
14
17
18
20
Case
1
11
15
17
2.1.2.2 Logistics and General Sampling Procedures
All 48 samples from the detailed surveys were collected during the
period from April 23 to May 29, 1984. The EPA furnished the vessel and
conducted all sampling operations except those scheduled in Sequim Bay.
Staff from the MRL were responsible for sampling Sequim Bay and used their
own boat and gear.
21
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Each sampling site was located with the precision navigational
system (Mini-Ranger); or in Sequim Bay, by sightings with a hand-bearing
O
compass. The samples were collected in two casts with a 0.1-m van Veen
grab sampler to obtain a 5-L composite sample. Excess seawater was then
decanted. Care was exercised to ensure a minimum penetration depth of 10
cm was achieved and that the samples were free of boulders and foreign
materials (tires, pieces of wood, rusted metal), and contained less than
20% gravel. Samples not achieving the minimum pentration depth, or
containing foreign materials were rejected and the cast retaken.
Rejection of the cast was appropriately documented. Cross contamination
of samples was controlled as described in Section 2.1.1.2.
As each grab sample was brought aboard the research vessel, three (3)
cores (5-cm diameter x 15-cm long) were taken from the undisturbed surface
of the sample for benthic infaunal analyses. The cores were gently
extruded into labeled sample jars containing buffered 10% formalin. Each
jar was capped and shaken to ensure thorough mixing of the preservative
with the sample.
After taking cores for benthic infaunal analyses, the remaining
sediment in each of the two grab samples was removed in vertical sections
from the top to the bottom of the sampler and placed into acid-cleaned,
prelabeled, 1-gal glass carboys. During this step, care was taken not to
collect any sediment that was in contact with the inside surfaces of the
van Veen sampler. The carboys were capped with appropriate liner
(aluminum or Teflon*), and iced. There was no mixing or further
subsampling in the field.
At the EPA Region 10 Laboratory, each sample was emptied into a
chemically cleaned glass aquarium and mixed manually for 20 min with an
acid-cleaned, hard plastic spoon. After mixing, subsamples were taken for
(1) priority pollutant analysis (minimum volume 1 L), (2) range-finding
chemistry (minimum volume 0.5 L), (3) amphipod bioassay (minimum volume
sample 3 L), and (4) oyster larvae bioassay (minimum volume sample 0.5 L).
EPA staff performed both mixing and subsampling.
* Registered trademark of the DuPont de Nemours Co., Inc., Wilmington,
Delaware.
22
-------
For four (4) samples collected in Sequim Bay, MRL staff were
responsible for transporting the samples to the Region 10 Laboratory for
mixing and subsampling.
2.2 PHYSICAL AND CHEMICAL (RANGE FINDING) SEDIMENT ANALYSES (PERFORMED BY
MRL)
2.2.1 Grain Size
Grain-size analysis was performed to estimate percent fractions for
gravel (>2 mm), sand (^2 mm >62 pm), silt (<62 pm >4 pm) and clay (<4 pm).
A wet weight sample of approximately 50 g was wet sieved through 2-mm and
62-pm screens. Material retained on the screens was dried (100°C for
24 h) and weighed to determine total sample weights of gravel and sand,
respectively. Material washed into the pan was transferred to a 1-L
graduated cylinder and distilled water was added to bring the total volume
to 1 L. The graduated cylinder was stoppered, and the sample was mixed by
inverting for 1 min and then allowed to stand for 29 sec. Ten-milliliter
pi pet samples were taken from 10 cm for silt plus clay and from 5 cm after
61 min and 29 sec for clay. These subsamples were dried (100°C for 24 h)
and the residue weighed. The weight of the residue was multiplied by 100
to calculate the total weight of silt and clay in the sample. Percent
contribution of each fraction was based on the aggregate weight of all
fractions.
2.2.2 Total Extractable Hydrocarbons Determined by Infrared
Spectrophotometry
2.2.2.1 Sampling
Twenty grams of sediment, 20 g of anhydrous sodium sulfate, and 25 mL
of carbon tetrachloride (CCLiJ were placed into a labeled 250-mL bottle
with a ground glass stopper. The bottle was placed on a shaker and
23
-------
agitated overnight (16 h). The CCLi* containing the extractable organic
material received infrared analysis using a modification of the procedure
described by Simard et al. (1951).
2.2.2.2 Analytical Procedures
Simard's procedure (Simard et al. 1951) was modified because
sediments rather than water were extracted. Analyses were run on a
Beckman1 Acculab 4 spectrophotometer by comparing the amount of absorbance
obtained at 2930 cm"1 to a calibration curve of Prudhoe Bay crude oil.
Recoveries of total oil from CCU extracts of oil-spiked sediments were
found to be >90%. Results were reported as ppm dry weight.
2.2.2.3 Calibration Procedures and Frequency
The infrared (IR) spectrophotometer was calibrated weekly using a
Beckman polystyrene wavelength/wavenumber calibrator for IR spectro-
photometers, Part No. 80785; Polystyrene film versus air.
2.2.3 Water Content
The sediment samples were stirred to homogenize, and then 20 to 50 g
were placed in a prepared 60-mL polystyrene bottle and weighed to the
nearest 10 mg. The samples were frozen to -80°C and freeze-dried
(10 millitorrs) to constant weight. Results were reported as percent
water, which was calculated as follows.
% water = — • 100
where A = wet weight
B = dry weight.
1 Beckman Instruments, Inc., Berkeley, California.
24
-------
2.2.4 Volatile Solids
Evaporating dishes were ignited for at least 60 min at a temperature
of no less than 550°C, then cooled in a desiccator, weighed to the nearest
10 mg and kept in the desiccator until ready for use. Five- to fifteen-
(5 to 15) gram aliquots of freeze-dried sediment were then placed in
evaporating dishes and weighed to the nearest 10 mg to establish sample
dry weights. The evaporating dishes and their contents were then ignited
for 60 min at 550°C, again cooled in a desiccator and reweighed to the
nearest 10 mg to establish the sample dry weights less the volatiles.
Results were reported as percent volatiles, which was calculated as
follows.
% volatiles = — • 100
where B = dry weight
C = furnace weight
2.2.5 Total Organic Carbon
Total organic carbon was determined using a Leco1 Model WR-12 carbon
analyzer. An 150- to 250-mg aliquot of dried sediment sample was
homogenized in a ceramic crucible. The sample was washed twice with
6N HC1 to remove carbonate carbon. If the reaction was vigorous on the
second wash, a third treatment with HC1 was conducted. Following
decarbonation, the sample was rinsed with distilled water until neutral
and dried at 45°C. The carbon analyzer was calibrated daily with freshly
prepared standards. Copper and zinc accelerators were added to the sample
crucibles and combusted with an induction furnace. The C02 evolved was
scrubbed of water, halide, and sulfur and the percent total organic carbon
was calculated. Typically, two duplicates and one blank were run for
every 25 samples.
1 Leco Corporation, St. Joseph, Missouri.
25
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2.2.6 Metals Analyses
2.2.6.1 Sample Handling
Sediment samples analyzed for trace metals consisted of 30-g aliquots
subsampled from we11-homogenized primary samples. Aliquots were taken
®
with a Teflon -coated spatula and placed into acid-cleaned polystyrene
jars. On return to the laboratory, the trace-metal aliquots were frozen
at -80°C in the jars until analysis.
Before analysis, the samples were freeze-dried to constant weight
with a Virtis1 Freeze-Mobile 12, and then homogenized on a Spex2 8000
mixer-mill. All of the samples were handled in the containers that they
were collected in to prevent contamination. This procedure has been shown
to yield no contamination for any trace metal measured.
2.2.6.2 Sediment Digestion Procedures
Before analyzing by atomic absorption spectroscopy, the samples were
digested in a 5:2:5 mixture of HNOa, HaSOi*, and HaO (Bloom 1983). This
procedure oxidized all organic matter and released all anthropogenic Pb,
Hg, and Ag. All digestates were diluted to 1.0 g of sediment (dry weight)
to 100 ml of solution. This dilution ensured greater reproducibility in
graphite furnace analysis and eased data reduction.
2.2.6.3 Atomic Absorption Procedures: Lead and Silver
Both Pb and Ag were measured using a Zeeman graphite furnace atomic
absorption spectrophotometer (GFZAAS). By using a state-of-the-art
Perkin-Elmer3 Zeeman background corrector and Model 5000
1 Virtis Co., Inc., Gardiner, New York.
2 Spex Industries, Inc., Edison, New Jersey.
3 Perkin-Elmer Corporation, Norwalk, Connecticut.
Registered Trademark of the Teflon Company.
26
-------
spectrophotometer, accurate measurements were provided even in such
complex matrices as digested sediments (Riley et al. 1980; 1981). The
detection limits, based on twice the standard deviation of triplicate
samples near the detection limit were as follows: Ag, 0.02 yg/g dry
weight, and Pb, 0.5 yg/g dry weight. These limits were at least ten times
lower than any values that were encountered in Puget Sound sediments.
2.2.6.4 Atomic Absorption Procedures: Mercury
Mercury was measured using the cold-vapor atomic absorption technique
in conjunction with dual cold traps (Bloom and Crecelius 1983). A
Laboratory Data Control1 ultraviolet monitor with 30-cm cell was used to
perform the analysis. With this technique, a detection limit of
0.005 yg/g dry weight was attained. This limit was ten times lower than
any value measured in Puget Sound sediments.
2.2.6.5 Calibration Procedures and Frequency
All atomic absorption calibration was done by the method of standard
additions according to manufacturer's instructions. A calibration curve
was prepared each day and all samples were analyzed under conditions that
gave a response in the linear range. Ten percent of the samples analyzed
were standards in the linear range. Standards were prepared from
commercially available (1000 mg/L) atomic absorption standards.
2.2.6.6 Detection Limits
Detection limits for metals in sediments were in accordance with EPA
guidelines and are presented in Table 4.
LDC/Milton Roy, Sunnyvale, California.
27
-------
TABLE 4. Detection Limits, Blanks, and Sediment
Standards Analyzed by MRL (ppm dry
weight)
Detection
Limits
0.02
0.005
0.02
Blanks
1.0
0.01
0.02
NBS 1646
MRL Value
22
0.074
0.072
Certified
Value
28
0.063
none
Element
Pb
Hg
Ag
2.2.6.7 Precision, Accuracy, and Completeness
The precision and accuracy of the methods were determined in
accordance with EPA guidelines for assessing and reporting quality for
environmental measurements. All other quality control (QC) data were
recorded and maintained in MRL laboratory record books.
2.2.6.8 Standard Materials (Accuracy)
Five percent of the samples analyzed were a standard material to
ensure the accuracy of the method. These materials included National
Bureau of Standards (NBS) estuarine sediment, National Research Council of
Canada (NRCC) estuarine sediment, and NBS air particulate matter.
Measured values deviated from the established value by more than one
standard deviation no more than 10% of the time. When this deviation
occurred, the parameters were reexamined and the standards rerun. If the
deviating value was repeated, in-depth investigation of the equipment and
sample were undertaken.
Note: For lead, the analysis may have consistently reported NBS
standard estuarine sediments 20% to 30% low because of the high
crustal/anthropogenic ratio in these clean sediments; therefore, we also
intercalibrated 5% of the enriched samples collected by nondestructive
x-ray fluorescence spectroscopy. These values were within ±10% of the
atomic absorption measurements.
28
-------
2.2.6.9 Blanks
Ten percent of the samples analyzed were procedural blanks. Blanks
for the three metals (Table 4) were consistently low and were routinely
subtracted from the sample concentrations. These values were always
recorded in MRL laboratory record books.
2.3 PRIORITY POLLUTANT SEDIMENT ANALYSES (PERFORMED BY EPA)
2.3.1 Procedures for Organic Compounds
Samples of thoroughly mixed wet sediments were dosed with surrogate
standards and extracted with acetone using a Soxhlet extractor. The
extract was concentrated, added to organic-free water, and extracted with
methylene chloride. This extraction was performed three times at pH 6 to
8 and three times at pH 2. The combined extracts were dried,
concentrated, and divided into equal portions for analysis of pesticide/
chlorinated hydrocarbon and semivolatile compounds.
2.3.1.1 Acid/Base-Neutrals
The portion to be analyzed using gas chromatography-mass spectrometry
(GC-MS) was further concentrated to 1.0 ml. The sample was analyzed with
a Finnigan1 5100 and Superlncos data system using a DB-5 fused silica
column and splitless injection. The gas chromatograph (GC) was held at
35°C for 2 min during injection and then programmed to 50°C at 5°C per
minute. After that, it was programmed from 50°C to 320°C at 8°C per
minute. Total time for analysis was 45 min. The mass spectrophotometer
scanned the mass range of 35 to 450 amu. The sample constituents were
quantified using the internal standard method. Six internal standards
were used for the entire range of the sample.
Finnigan MAT, San Jose, California.
29
-------
The internal standards were d^-l^-dichlorobenzene, d8-naphthalene,
D10-acenaphthalene, d10-phenanthrene, d12-chrysene, and d12-benzo-a-
pyrene. All except the last compound are listed in the Statement of Work
(SOW) for Organics Analysis printed by the EPA Contract Laboratory
Program. The procedure for using internal standards for quantification is
described in the SOW and also in Method 625 of the Federal Register,
Vol. 49, No. 209, October 26, 1984.
2.3.1.2 Pesticides/PCBs/PCBDs
The solvent for the pesticide, PCB (polychlorinated biphenyl) and
PCBD (polychlorinated butadiene) determinations was changed from methylene
chloride to petroleum ether. The sample volume was reduced to 1.0 ml by
volatilization of the solvent with nitrogen gas, and then placed on a
Florisil column. Two hundred milliliters of pentane were used to elute
PCBs, PCBDs, and DDE. A 200-mL mixture of equal volumes of diethyl ether
and petroleum ether was used to elute the remaining pesticides. The
volume of each eluate was reduced to 1.0 ml.
The EPA Pesticides and Industrial Chemicals Repository at Research
Triangle Park, North Carolina, is the source for all pesticide, PCB and
PCBD standards used in this study.
The PCBs were identified and quantified based on the calculation of
peak height of all major peaks (at least 6 to 8) and comparison to a PCB
standard. When evidence indicated that erroneous data would result from
the interference of extraneous compounds such as DDT analogs, only those
peaks that were free of such interferences were used for quantification.
The Manual of Analytical Methods for the Analysis of Pesticides in
Humans and Environmental Samples (EPA-600/8-80-038), 1980, Section 9 was
used as the reference.
For the PCBD analysis, 5 yL of the pentane fraction were injected
on a 3% OV-225 column at 40°C, isothermally. The detector used for
30
-------
quantification was a Hall 700A Microconductivity Detector. Confirmation
was achieved using a 1% OV-1/2.4% OV-225 column at 40°C isothermally with
a 63Ni electron capture (EC) detector. Chromatography was completed in
15 min.
A qualitative standard containing PCBDs was obtained from NOAA. Peak
identification/assignments were determined by GC-MS using a 3% OV-225
packed column which is the same column used for analysis with the Hall
700A microconductivity detector. The Hall detector was calibrated using
hexachlorobutadiene and equivalent response factors were determined for
trichloro-, tetrachloro-, and pentachlorobutadiene.
For the regular pesticide/PCB analysis, the primary column was a
6-ft x 2-mm packed column that contained 1.5% SP2250/1.95% SP3401 as the
stationary phase. The secondary column contained 4% SE-30/6% OV-210. The
temperature of both columns was 200°C. Chromatography was completed in
30 min. The primary detector used was a 63Ni EC detector, and the
confirming detector was a 3H EC detector. The GC used was a Tracer1
Model 222. Three levels of standards were injected daily before samples
were analyzed. Percent recoveries of o, p'-DDE, the surrogate spike, were
calculated, but the results were not adjusted by these recoveries.
2.3.1.3 Gas Chromatography/Mass Spectrometry Analysis-Volatile
Organic Analysis (VOA)
Approximately 2 g of sediment were added to a 16-mm x 125-mm screw-
top culture tube that had previously been cleaned in a muffle furnace at
410°C. The tube was sealed with a screw cap containing a Teflon liner,
and stored at 4°C until analysis. Just before purging, 5 ml of
organic-free water^ containing the internal and surrogate standards, were
added to the sample in the culture tube. The volatile organic analyses
were spiked with 50 ng of each compound, except for acrylonitrile, which
was spiked with 100 ng. The tube was connected to a Tekmar2 LSC-3 purge
Hall is a registered trademark of Tracer Inc., Austin, Texas.
1 Tracer, Inc., Austin, Texas.
2 Tekmar Company, Cincinnati, Ohio.
31
-------
and trap instrument and sparged onto a trap containing one-half Tenax and
one-half coconut charcoal in series. The volatile compounds were desorbed
for 4 min at 180°C into a DB-5, fused silica, capillary column (1.0-y/m
film thickness, 30-m long). The flow was split in a 1:10 ratio. A loop
of the column at the front portion was immersed in a thick slurry of solid
powdered carbon dioxide and methanol at least 1 min before desorption, and
kept there for the entire desorption period. The GC-MS was initiated at
the same time as desorption. The C02/methanol cold trap was removed
immediately after desorption and the GC oven kept at 30°C for an
additional 6 min. The oven was then programmed to increase from 30°C to
120°C at a rate of 6°C per min and held for 2 min or until the final
compound, 1,4-bromofluorobenzene, had eluted. The analysis was performed
on a Finnigan GC-MS 3200 with an INCOS data system.
2.3.2 Procedures for Metals
The sediment samples were dried at 105°C, pulverized and screened
through No. 20 mesh or equivalent nylon screen. One gram of sample was
digested with a mixture of nitric acid and hydrogen peroxide. The sample
was filtered through a 0.8-ym membrane filter, and the volume was adjusted
to 100 mL.
The metals were determined by atomic absorption spectrophotometry
using a Zeeman flame furnace following EPA methods. Mercury was
determined on a wet sample by the manual cold vapor technique according to
EPA Method 245.5. Wet weight values were transformed into dry weight
values on the basis of percent solids data.
The mean concentrations of Ag, Hg, and Pb in sediments collected in
1983 and 1984 were determined by MRL. The concentrations of these same
metals were also determined by the EPA Region 10 Laboratory and both sets
of results are presented in Appendix B (Tables B-9 to B-16). For the most
part, results from the two laboratories compare well; however, a few
discrepancies exist (e.g., Hg at Fourmile Rock - Elliott Bay dump site
vicinity; Ag in Sinclair Inlet). Unfortunately, these few discrepancies
cannot be resolved because the EPA Laboratory did not analyze standard
reference sediments.
32
-------
2.3.3 Procedures for Solids (Percent)
Samples were dried at 103°C to 105°C according to EPA methods.
2.3.4 Procedures for Phenolics and Cyanides
Phenolics and cyanides were distilled and the distillate analyzed by
automated spectrophotometry following EPA methods.
2.3.5 Detection and Quantisation Limits
The limit of detection of a compound is defined as an analyte signal,
three standard deviations above the peak-to-peak noise level of the
instrument in the region of the expected analyte peak. Any signal not
exceeding this minimum response would be judged as not indicating the
presence of the analyte. The limit of quantisation is defined as the
analyte signal that is 10 standard deviations above the noise level of the
instrument. Between these two limits is a region of detection in which a
compound can be identified, but where the concentration is too low to
allow an accurate estimate of its value.
The reviewer should refer to "Guidelines for Data Acquisition and
Data Quality Evaluation in Environmental Chemistry," Anal. Chem. 52, p.
2241 (1980).
In Appendix B (Tables B-17 to B-26), when the symbol V is used, the
value preceding it is the quantisation limit for the particular substance.
It means that the concentration, if any, falls below the detection limit.
The symbol "m" indicates that the concentration of the substance falls
between its detection limit and its quantisation limit.
The quantisation limits (QL) of the acid/base-neutral compounds are
given as a range. The quantisation limits are not all the same because
the samples had a varying percent solids value. A high percent solids
means a lower quantisation limit and vice versa. Also the ranges are not
all alike because some compounds are more sensitive to detection than
others. Most of the VOA compounds had a similar quantisation limit, so
that an average was given for them. The range of the QL for all but
acrolein and acrylonitrile was 4.1 to 18 ug/kg, dry weight. The
33
-------
pesticides were also affected by the percent solids value, but the
variation was always between 0.3 and 1.0 yg/kg. Therefore, their QL is
reported to the nearest whole number.
The quantitation limits of all target organic compounds are listed in
Table 5. The quantitation limits of the VOA compounds are generally lower
than the acid/base-neutral compounds because the purge and trap procedure
eliminates many matrix problems. Detection limits for metals are reported
in Table 6.
2.3.6 Precision, Accuracy, and Completeness
The accuracy of the methods was determined in general accordance to
the EPA Handbook for Analytical Quality Control in Water and Wastewater
Laboratories (1979). The average recovery and standard deviation (CT)
for many spiked samples was determined, and the control limits were
calculated to be ± 3CT about the mean. The Statement of Work for Organics
Analysis for the EPA Contract Laboratory Program (CLP) (Rev. Jan. 1985)
outlines a detailed QA procedure, but unfortunately,'it was not available
at the time this survey was performed. For the purpose of comparison of
QC limits, CLP-suggested spiking compounds that were used for the QA of
this study will be examined more closely.
2.3.6.1 Organic Parameters
The quality of the Puget Sound sediment data was determined with the
use of quality control and surrogate spiking compounds. Table 7
summarizes the scope of the QC work.
2.3.6.2 VOA Fraction
The surrogate spiking compounds used were 1,2-dichloroethene-d,^
d6-benzene, d8-toluene, and 1,4-bromofluorobenzene. The average recovery
of each surrogate compound is presented in Table 8.
34
-------
TABLE 5. Average Quantitation Limits of Organic Compounds
Analyzed in Puget Sound Sediment Samples
Organochlorine Compounds (ppb)
Aldrln
Chlordane
Dieldrln
4,4'-DDT
4,*'-DDE
4,*'-ODD
Alpha Endosulfan
Beta Endosulfan
Endosulfan Sulfate
Endrin
Endrln Aldehyde
Heptachlor
Heptachlor Epoxide
Alpha BHC
Beta BHC
Lindane
Delta BHC
Toxaphene
1 Hexachlorobenzene 1
1 Polychlorlnated Butadiens 2
1 PCB 1016 20
1 PCB 1221 20
1 PBC 1232 20
1 PCB 1242 20
1 PCB 1248 20
1 PCB 1254 20
30 PCB 1260 20
OJ
in
Acenaphthene 40-200
1,2,4-Trichlorobenzene 80-200
Hexachlorobenzene 400-1000
Hexachloroethane 100-250
bis (2-Chloroethyl)ether 40-200
2-Chloronaphthalene 40-200
1,2-Dlchlorobenzene 40-200
1,3-Di chlorobenzene 40-200
1,4-Dfchlorobenzene 40-200
2,4-Dlnltrotoluene 100-500
2,6-Dlnitrotoluene 100-500
1,2-Dlphenylhydrazine 100-250
(as azo benzene)
Fluoranthene 40-250
Base Neutral Compounds (ppb)
4-Chlorophenylphenylether 100-250
4-Broinophenylphenyl ether 200-500
bi s(2-Chloroi sopropy1)ether 40-200
bis(2-Ch1oroethoxy)methane 40-200
Hexachlorobutadiene 160-400
Hexachlorocylopentadlene 600-1500
Isophorone 40-200
Naphthalene 40-200
Nitrobenzene 40-200
N-Ni trosodiphenylami ne 1000-2500
N-Ni trosodi-n-propy1 ami ne 600-1500
bi s(2-Ethylhexyl)phthalate 100-1 TOO
Butyl benzylphthalate 40-200
Di-n-butylphthalate 40-200
Benzidine 1000-2500
3,3'-Dichlorobenzidine 100-500
Dibenzofuran 40-100
Diethylphthalate 40-200
Dimethylphthalate 40-200
Benzo(a)anthracene 40-100
Benzo(a)pyrene 100-250
Benzo(k)fluoranthene 40-100
and/or Benzo(b)
Chrysene 40-100
Acenaphthylene 40-200
Anthracene 40-200
Benzo(ghi)perylene 200-800
Fluorene 40-200
Phenanthrene 40-100
Dibenzo(a,h)anthracene 200-800
Indeno (1,2,3-c,d)pyrene 200-800
Pyrene 40-100
Benzyl Alcohol 160-400
2-Methylnaphthalene 40-200
*Dry weight basis.
-------
TABLE 5. Average Quantitation Limits of Organic Compounds
Analyzed in Puget Sound Sediment Samples
(Continued)
Acid Compounds (ppb)
2,4,6-Trlchlorophenol 200-500
p-Chloro-m-cresol 100-250
2-Chlorophenol 40-200
2,4-Dlchlorophenol 200-500
2,4-D1methyl phenol
2-N1trophenol
4-Nitrophenol
2,4-Dlnltrophenol
2-Methylphenol
2,4,5-THchlorophenol
100-250
200-500
600-1500
1000-2500
100r250
200-500
4,6-D1n1tro-o-cre»ol 1000-2500
Pentachlorophenol 600-1500
Phenol 40-200
Benzole Acid 600-1500
4-Methylphenol 100-250
Ol
Acrolein 240
Acrylonltrlle 120
Benzene 12
Carbon Tetrachloride 12
Chlorobenzene 12
1,2-D1chloroethane 12
1,1,1-Trichloroethane 12
1,1-D1chloroethane 12
1,1,2-TMchloroethane 12
1,1,2,2-Tetrachloroethane 12
Acetone 12
4-Methyl-2-Pentanone 12
o-Xylene 12
VOA Compounds (ppb)
Chloroethane 12
Chloroform 12
1,1-Dlchloroethylene 12
1,2-trans-Dichloroethylene 12
1,2-Dlchloropropane 12
c1s-1,3-D1chloropropene 12
trans-1,3-D1chloropropene 12
Ethyl benzene 12
Methylene Chloride 12
Methyl Chloride 12
2-Butanone 12
Carbon Disulfide 12
Methyl Bromide 12
Bromoform 12
Bromodichloromethane 12
Trlchlorofluoromethane 12
Dlbromochloromethane 12
Tetrachloroethylene 12
Toluene 12
Trlchloroethylene 12
Vinyl Chloride 12
2-Chloroethyl Vinyl Ether 12
2-Hexanone 12
Styrene 12
-------
TABLE 6. Detection Limits of Metals in Sediments (1.0 g/100 ml)
Element
Antimony
Arsenic
Beryllium
Cadmium
Chromium
Cobalt
Copper
Lead
Mercury
Nickel
Selenium
Silver
Thallium
Zinc
Detection Limit ppm (dry weight)
0.1
0.1
0.02
0.02
0.05
0.05
0.05
0.02
0.001
0.05
0.1
0.01
0.1
0.2
TABLE 7. Quality Control Samples Analyzed
Fraction
Acid/Base-Neutral
VOA
Pesticide/PCB
Samples
Analyzed
48
48
48
Blanks Duplicates Spikes
4
13
4
1
0
1
5
5
3
Duplicate
Spikes
4
5
3
37
-------
TABLE 8. Average Percent Recovery of Surrogate
Compounds of VOA Fraction
1,2-Dichloro- 1,4-Bromo-
Type ethene-d^ d6-Benzene d8-Toluene fluorobenzene
Sample 102.7 ± 13.0 97.1 ± 13.4 100.0 ± 15.5 73.0 ± 15.0
Blanks 96.5 ± 13.6 100.8 ± 10.5 102.8 ± 11.2 95.0 ± 13.4
Matrix Spikes 105.3 ± 22.3 101.8 ± 5.1 99.4 ± 7.0 76.0 ± 11.3
The internal surrogate standards were added to 5 ml of organic-free
water, which was then added to the sediment sample just before sparging in
the gas purge device.
All of the VOA compounds were in the matrix and duplicate matrix
spikes. They were added to the organic-free water that was mixed
with sediment before sparging. Table 9 lists the total recovery for all
compounds as well as the five VOA compounds recommended by CLP, and the
Region 10 QC limits.
Table 9 shows that all but one of the recovery data for all compounds
and the five CLP compounds were within the Region 10 control limits. Only
two compounds were not within CLP control limits, but they were for
different compounds in different samples.
The number of duplicate spikes was insufficient to prepare a
Region 10 control chart for precision. However, the relative percent
difference (RPD) of two recovery values from the CLP could be used to
obtain an approximate measure of precision. The RPD is calculated as
follows:
D1 - D2
RPD = — x 100
(D! + D2)/2
where DI = first spike value
and D2 = second spike value (duplicate).
38
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TABLE 9. Percent Recoveryof VOA Spikes
Sample No.
17008MS
17008MSD
19058MS
1 9058MSD
20057MS
20057MSD
21057MS
21057MSD
22058MS
22058MSD
CLP Recovery
QC Limits
Region 10
Recovery
QC Limits
Average
Recovery
Matrix Spike
92.8
96.5
114.4
105.4
101.7
114.5
122.6
125.5
109.9
132.3
-
*
67-151
CLP-Suggested
1,1-Dichloro-
ethene
69
89
137
107
88
103
139
145
57
82
59-172
13-178
Trichloro-
ethene
95
95
92
96
104
94
121
132
112
121
62-137
70-136
Compound Recovery
Chloro-
benzene
89
87
84
90
80
82
88
95
83
103
60-1 33
50-127
Toluene
138
162
92
97
104
100
116
124
101
122
59-139
48-159
Benzene
107
109
100
104
104
101
125
140
106
122
66-142
53-163
'''CLP does not give control limits for the above parameter.
The precision of the spikes is shown in Table 10.
TABLE 10. Precision of VOA Spikes (RPD)
Sample No.
17008
19058
20057
21057
22058
CLP RPD
QC Limits
1 ,1-Dichloroethene
25
24
16
4
52
22
Trichloroethene
0
4
10
9
8
24
Chlorobenzene
2
7
2
8
21
21
Toluene
16
5
4
7
19
21
Benzene
2
0
3
11
14
21
39
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From Table 10, values for 1,1-dichloroethene seem to be high, but the
other compounds indicate that the precision of the spikes appear to be
valid, and, except for one point, the dichloroethene problem is not too
severe.
The VOA blanks were generally free of halogenated compounds. The
only compounds that appeared regularly were acetone and methylene
chloride. Acetone concentrations were generally 10 yg/kg or less, and the
CH2C12 concentrations were 1 yg/kg or less. Other compounds present were
2-butanone, 2-hexanone, 4-methyl-2-pentanone, toluene, ethyl benzene, and
ortho-xylene. These compounds were not present in more than 40% of the
blanks and concentrations were 20 yg/kg or less.
2.3.6.3 Acid/Base-Neutral Fraction
Table 11 gives the results of the average recoveries and percent
relative standard deviations (% RSD) of the surrogate standards in all of
the samples, blanks, and spikes.
TABLE 11. Average Recovery of Acid/Base-Neutral Surrogate Standards
d5-Phenol Pentafluorophenol d5-Nitrobenzene d10-Pyrene
Mean Percent
Recovery 77.7% 48.9% 63.0% 43.9%
o ± 26.8% ± 28.4% ± 23.2% ± 14.9%
Percent RSD 22% 59% 38% 34%
Table 12 lists the recovery of the acid and base-neutral compounds in
the matrix spikes. The samples were spiked with all 11 acid compounds,
but some of the base-neutral spiking solutions were depleted during the
sampling and analysis period, and could not be replaced. At that time,
the EPA Laboratory was also in the process of switching to new spiking
standards. Ten acid and base-neutral spiking compounds that were
available for the whole year are presented in the table. The average
40
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TABLE 12. Matrix Spike Percent Recovery of the Acid/Base-Neutral Fraction
Region 10 CLP
Matrix Spike
Compound
Average recovery
Acid & B/N cmpds.
Average recovery
Acid compounds
4-Chloro-3-
Methyl phenol
2-Chlorophenol
4-Nitrophenol
Pentachl orophenol
Phenol
1,2,4-Trlchloro-
benzene
1,4-Dlchloro-
benzene
Hexachloro-
butadlene
Nitrobenzene
Di ethyl phthalate
17008Y
137
120
128
99
470
0
102
99
56
46
78
80
1 7008YJ
120
120
118
102
430
15
99
103
73
78
54
58
18050Y
56
66
94
34
0
59
57
41
0
12
22
73
1 8050YJ
56
59
81
8
155
72
31
0
0
0
0
61
1 9059Y
89
97
141
76
119
22
77
99
51
50
74
82
20059Y
59
54
71
78
0
65
88
106
65
49
77
57
20059YJ
74
76
84
87
93
62
85
108
63
47
77
57
22051Y
86
90
82
78
183
39
75
105
60
60
86
79
22051 YJ
87
91
88
81
247
29
72
104
61
62
90
87
Control
Limits
0-180
7-157
0-218
0-171
0-456
0-119
0-192
0-190
0-134
0-141
0-167
0-159
Contr<
Limit!
1
1
26-103
25-102
11-114
17-109
26-90
38-107
28-104
1
1
1
CLP does not give control limits for these parameters or compounds.
-------
recovery of all compounds, and the acid compounds only, in each matrix
spike is presented in the table. The average recoveries for all compounds
are not an accurate indicator of precision, because the number and kinds
of base-neutral compounds varied over the analysis period. The acid
compound recoveries accurately represent the spiking of each matrix spike.
The Region 10 Laboratory control limits were ± 3o about the mean
recovery. The control limits are not symetrical about the mean because,
most of the time, the value of 3o exceeded the mean. Therefore, the
lowest recovery limit used was 0%.
When the Region 10 control limits are used, 10 values out of 90, or
11% of the values, are at, or beyond, the limits, including 8 compounds
that had 0% recovery. When the CLP control limits are applied, 21 out of
the same 90 values, or 23% of the values, are beyond the CLP limits. Two
reasons could explain why more sample values are beyond the CLP limits.
First, the CLP range is narrower than the Region 10 range, indicating that
perhaps a larger number of values were used when calculating the average
or the values used were more precise. Second, the CLP values indicate low
average recoveries. The largest recovery possible for the acid compounds
that still is within the limits is 114%. Some of the spike values were
not unreasonably higher than 100%, but were still out of the CLP range.
The compound that gave the most difficulty was 4-nitrophenol. The
Region 10 control limits for this compound are unrealistically large, as
are some of the listed recoveries. The extraction procedures should be
closely followed to be sure that they are not the cause of the variation.
Matrix spike duplicate 18050YJ had four values each beyond Region 10 and
CLP limits, including four base-neutral values that were zero. Its
duplicate had values that were better, so the problem probably was not due
to matrix effects.
The number of duplicate spike pairs was insufficient to calculate
Region 10 RPD limits for precision. The CLP limits were used as
indicators of precision, and the results obtained are given in Table 13.
The samples that show an RPD of 200 are those in which one of the
duplicate recovery values is zero. Nitrophenol, again, appears
to be a recurring problem, and the spikes of 18050 also appear to be
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unsatisfactory. The zero RPD for 1,4-dichlorobenzene for 18050 is not
meaningful, because the percent recovery for this compound in both
duplicates is zero. A total of 7 RPD values out of 28, or 25%, are
unsatisfactory.
TABLE 13. Precision of Acid/Base-Neutral Spikes (RPD)
Sample No.
Compound
4-Chloro-3-methyl phenol
2-Chlorophenol
4-Nitrophenol
Pentachlorophenol
Phenol
1,2,4-Trichlorobenzene
1,4-Dichlorobenzene
17008
8
3
9
200
3
4
26
18050
8
124
200
20
59
200
0
20059
17
11
200
5
3
2
3
22051
7
4
30
29
4
1
2
CLP RPD
Limits
33
50
50
47
35
23
27
Four acid/base-netural solvent blanks were prepared simultaneously
with the samples. Except for occasional phthalates, the blanks were free
of detectable amounts of target compounds.
2.3.6.4 Pesticide Fraction
The pesticide recovery efficiency was monitored by using a, p'-DDE as
a surrogate standard. Some samples contained high levels of PCBs that
interfered with the quantisation of the pesticide surrogate spike. But
for the 38 samples, spikes and blanks that did have quantifiable a,
p'-DDE, the average recovery was 91.6% ± 15.8%. The CLP suggests
dibutylchlorendate as the surrogate standard, with an acceptable range of
46% to 136%. If a similar range was assumed to be reasonable for a,
p'-DDE, then the average surrogate recovery is satisfactory. In fact, all
individual surrogate recoveries were in this .range.
One of the samples, 21056, was analyzed in duplicate. No detectable
amount of pesticide or PCB was found in either duplicate.
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Three other samples were spiked with known amounts of pesticide
compounds. Unfortunately, two of these spikes had high levels of PCB that
interfered with the recovery quantisation. As a consequence, only one
sample was effectively spiked. For all 16 pesticide compounds, the
percent recoveries of the duplicates were 117% ± 25% and 116% ± 18%. The
CLP recommends six pesticides to monitor spike recovery. The recommended
recovery range of the CLP compounds averages from 35% to 133%. The
average recovery of all 12 values of the duplicates was 116%. Two of the
values, both for 4,4'-DDT, were one percentage point beyond the
recommended upper limit. All of the other individual values were within
their specific range. Precision between duplicate values was good. The
RPD for the six pesticide compounds ranged from 31% to 50%, with an
average of about 43%. None of the RPDs for each pesticide spiking
duplicate exceeded this range of its specific RPD limit. The RPD of each
pesticide was, in fact, always one-half or less of its limit. Although
the Standard Reference Material for "PCBs in sediment" were available,
they were not used, because of an oversight. (They have been used
successfully in the past.)
2.4 BENTHIC INFAUNAL ANALYSES (PERFORMED BY MRL)
2.4.1 General Procedures
Preserved samples (six cores per station, each core 5 cm x 15 cm)
were placed in enamel pans and a quantity of freshwater was added. The
contents were gently agitated. The water and suspended organisms were
then poured through a 1.0-mm mesh screen, and the organisms retained were
transferred to a water-filled petri dish. This "panning" process tended
to break down large clumps of sediment and suspended relatively light and
fragile organisms such as gammarid amphipods and juvenile polychaetes.
This process was usually repeated six times to remove virtually all types
of organisms from the sediment with minimum damage. Sediments remaining
in the pan were then placed in a 1.0-mm mesh sieve and washed with
freshwater, entraining the heavier organisms such as bivalves and
tube-dwelling polychaetes on the screen.
All organisms from a sample were then placed in freshwater in a petri
44
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dish, examined under a dissecting scope, and sorted into major taxa.
Common, easily identified species were enumerated at this time. More
difficult groups were set aside for more detailed examination. All
specimens were transferred to 70% isopropyl alcohol.
Each major taxonomic group was identified to the generic/specific
level if possible, but at the discretion of the taxonomic specialist. If
organisms were damaged or immature, identification was to the lowest
possible taxonomic ranking. Organisms were identified using stereo
(dissecting) and compound microscopes. Identification and enumeration
were always conducted in the "blind"; that is, the specialist conducting
the analyses did not know the origin (bay, station) of the particular
sample being quantified. And, as an additional quality assurance measure,
the taxonomic identifications from 10% of all benthic samples (5 of 48
stations) were verified by a second (independent) specialist, Mr. Jack Q.
Word of Evans-Hamilton, Inc. After being identified all samples were
stored in 70% isopropyl alcohol with approximately 20% glycerol added to
prevent desiccation.
The general taxonomic keys of Kozloff (1976), Smith and Carlton
(1975), Hobson and Banse (1981), and Banse and Hobson (1974) were
used together with specific supplementary publications for precise
identifications (i.e., Austin and Haylock 1973; Holleman 1972; McCauley
and Carey 1967).
2.4.2 Statistical Treatment
Infaunal data were tabulated by abundance (number of individuals per
station) and richness (number of taxa per station). Correlation analyses
(see Section 2.8) were performed with these data to determine
relationships with physical and chemical properties of the sediments.
2.5 FISH AND INVERTEBRATE PATHOLOGY (PERFORMED BY NOAA)
2.5.1 Fish Capture and Necropsy
The target fish species, English sole (Parophrys vetulus) and Dover
sole (Microstomus pacificus) were collected from the RV Harold W. Streeter
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with an otter trawl having a 7.5-m opening, a 10.8-m total length, 3.8-cm
mesh in the body of the net, and a 0.64-cm mesh liner in the cod end.
Individual trawls were for 5 min and covered a distance of approximately
0.2 nautical miles. At each sampling station (Table 1), English sole were
sorted first by gender; then each group was separated into five size
classes. When possible, up to six individuals of each size class, up to a
total of 30 fish, were selected for necropsy. English sole less than
150 mm in total length (0+ year class) were not sampled because this age
group previously has been found to possess no neoplastic lesions and very
few idiopathic lesions of the liver.
Total sample size at each station was determined on the basis of
differences in incidences of neoplasms determined in previous studies of
Puget Sound. Because neoplasms are relatively rare among the important
idiopathic lesions of interest, detecting significant differences in the
incidence of this lesion would require a large sample size. Fish from
significantly contaminated sites, such as the Duwamish River, have been
found to exhibit neoplasms in more than 15% of the population sampled,
while fish from reference areas have been found to have a very low (near
0%) incidence of neoplasia. For a two-way comparison of incidence
(a = 0.05, power = 0.80, two-tailed test), at least 30 fish are required
to detect significant differences between an incidence rate near zero and
incidence rates greater than or equal to 15%. Consequently, a total
sample size of 30 fish per site was selected for necropsy.
Fish to be necropsied were measured for total length (mm), weighed
(g) and assigned an individual field number. The spinal cord was then
severed and otoliths were collected to determine fish age (year-class).
The gender was determined and tissues were excised from the following
organs: liver, kidney, spleen, gall bladder, small intestine, gonad,
heart, and gill. Any grossly visible lesions were noted and recorded and
tissue specimens were collected. Tissues were placed in labeled cassettes
and immersed immediately in Dietrich's fixative (Gray 1954).
46
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2.5.2 Fish Histologlc and Diagnostic Procedures
Tissue specimens were stored in fixative for a minimum of 48 h, then
they were dehydrated and paraffin-infiltrated with an automated tissue
processor, embedded in paraffin and sectioned at a thickness of 5 to 6 ym.
Sections were routinely stained with Mayer's hematoxylin and eosin-
phloxine (Armed Forces Institute of Pathology 1968). All slides were
examined by light microscopy employing a "blind" system, in which the
examining histopathologist had information only on the species, length,
weight, sex, and description of grossly visible lesions, with no knowledge
of the location from which the fish was collected. Diagnoses were then
performed and the diagnostic information was coded on National Ocean Data
Center (NODC) File type 13, and placed on computer tape along with
information on gross pathology and other biological characteristics of
each fish. Data management and lesion incidence analyses were performed
using Minitab version 81.1 (Ryan et al. 1981) and SPSS (Statistical
Program for Social Sciences) versions 8.1 and 9.1 (Nie et al. 1975) on a
Burroughs 7800 computer.
2.5.3 Crab and Shrimp Capture and Necropsy
Crab (Cancer magister and Cancer gracilis) and shrimp (Pandalus
platyceros and Pandalopsis dispar) were collected using methods identical
to those used for target fish species at sampling stations shown in
Table 2. Some crab were also caught with baited crab ringnets set in the
sampling areas. Crab and shrimp to be necropsied were examined for
externally visible anomalies. Crab were measured for carapace width and
shrimp for carapace length (mm).
Some specimens of both crab and shrimp were necropsied immediately
after capture, while others were held in seawater-filled ice chests until
they could be processed. Necropsy of crabs was initiated by clipping off
all walking legs and then carefully separating the body carapace from the
underlying epidermis, enabling the prosector to then remove the internal
organs. Tissues were then excised for examination: antennal gland,
bladder, cardiac and pyloric stomachs, epidermis, esophagus, eye, gill,
47
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gonad, hemopoietic tissue, hepatopancreas, hindgut, midgut, and midgut
ampulla. Crab tissues were fixed in Helly's fixative (Humason 1979).
Shrimp were first injected by syringe with Davidson's fixative
(Humason 1979) directly into the hepatopancreas and body cavity. The
legs, antennae, and abdomen were then cut off and the carapace split open
to expose the internal organs. Whole shrimp were then placed in ,
Davidson's fixative. Tissues taken for examination were: antennal gland,
bladder, cardiac and pyloric stomach, epidermis, esophagus, eye, gill,
gonad, heart, hepatopancreas, hemopoietic tissue, midgut, and nervous
tissue.
2.5.4 Crab and Shrimp Histological and Diagnostic Procedures
Crab and shrimp tissues were fixed for 48 to 72 h and then stored in
70% ethanol until processed in the same manner as described for fish
tissues. Tissues were routinely stained with Harris1 hematoxylin and
eosin-B-phloxine (Armed Forces Institute of Pathology 1968). All tissues
were examined by light microscopy using the same "blind" system as used to
examine fish tissues.
2.5.5 Statistical Treatment
The 6-statistic (Zar 1974; Sokal and Rohlf 1969) was used to test for
differences in lesion incidence between stations within each size class of
English sole. For other fish and invertebrate species, the G-statistic
was used to simply identify differences in lesion incidence between
stations. In cases where the incidence patterns were significantly
heterogeneous (p <0.05), the individual areas or stations contributing to
the heterogeneity were identified by inspection.
2.6 AMPHIPOD BIOASSAY (PERFORMED BY EPA)
2.6.1 Strategy
The acute toxicity of Puget Sound sediments to the marine amphipod,
Rhepoxynius abrom'us, was determined by the general method of Swartz
48
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et al. (1984). This approach involved exposing amphipods to test and
control sediments for 10 days at 15°C, under continuous illumination and
aeration. Sediment toxicity was based on measurements of amphipod
survival, emergence, and reburial.
2.6.2 Test Organisms and Control Sediments
Amphipods and control sediments were collected weekly from West
Beach, Whidbey Island, Washington using a stainless steel benthic
dredge. The amphipods were kept in their natural sediment substrate
(control sediment) and transported on ice to the EPA Region 10 Laboratory,
Manchester, Washington, the day of collection. At the laboratory,
the amphipods were placed in a flow-through seawater system and slowly
acclimated to water temperatures (5°C per day) approaching the test
temperature (15°C). Control sediments were obtained by first wet sieving
West Beach sediments through a polypropylene screen having a mesh size of
1.0 mm. The sediment was then wet sieved through a 0.5-mm mesh,
polyethylene screen into seawater (salinity, 28.0 °/00) and kept at 4°C
until distributed for testing.
2.6.3 Screening Surveys
Using a sediment toxicity screening approach similar to the one
followed by Swartz et al. (1982) in Commencement Bay, test sediments (with
the exception of control sediments) collected during screening surveys
(August 2 - September 18, 1983) were assayed without replication. Control
sediments were assayed using five laboratory replicates. While testing
single replicates limited the statistical treatment of the data generated,
this approach provided the broadest screening coverage achievable within
the time allotted.
Three hundred grams (wet weight) of well-mixed sediment were added to
1-L glass beakers and brought up to 850 ml with filtered, Clam Bay
seawater. Interstitial water salinities associated with each sediment
sample were measured with a refractometer before the sediments were added
to the test beakers. The salinity of the seawater used to overlay the
49
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sediments was adjusted to 28.0 °/00 with distilled-demineralized water
before filtration.
Sediment and seawater preparations were covered with watch glasses
and plastic wrap, then incubated overnight (without amphipods) at 15°C
under continuous aeration and "cool white" fluorescent illumination
(10-60 ft-c). The next day, 20 amphipods were randomly added to each test
beaker and the contents of the beakers brought up to 950 ml. The "seeded"
beakers were then incubated for 10 days as described above.
The pH, dissolved oxygen concentration, and salinity of the
overlaying water were measured for small samples collected from each
sediment preparation just before adding the amphipods and again at the end
of the test. Floating amphipods were counted daily then gently tapped
down with a pipet tip. Amphipods surviving the 10-day exposure period
were counted and their ability to rebury in control sediment determined.
2.6.4 Detailed Surveys
Those test sediments collected during detailed surveys (April 23 -
May 29, 1984) were assayed using five laboratory replicates. Otherwise,
all methods were applied as described in the previous section.
In addition to the routine assay of non-toxic control sediment, the
response of each lot of amphipods to a reference toxicant was also
measured. This approach was used to determine the potential for a "batch"
effect on our assay. The toxicant, sodium pentachlorophenate (PCP), was
included in the test protocol in an effort to detect changes in the
sensitivity of the amphipods during the 6-week sediment testing period.
Sealed ampules of PCP were obtained from the U.S. Environmental Protection
Agency, Environmental Monitoring and Support Laboratory, Cincinnati, Ohio.
(Sodium Pentachlorophenate; Toxicant Concentrate #4; 3210 mg/L; Ampule
Serial Numbers 153, 157, 159, 170, 174, 176; October 15, 1979.) Fresh PCP
concentrations (0.1, 0.2, 0.3, 0.4, and 0.5 mg/L) and a control (0.0 mg/L)
were prepared in seawater just before each assay. The seawater diluent
was drawn from the same batch of seawater used to overlay the sediment
preparations described earlier. The sediment-free test preparations were
assayed in triplicate by first adding 200 mL of test solution to each of
three 250-mL glass beakers. Each beaker was then randomly seeded with 10
50
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amphipods, loosely covered with a plastic petri dish, and incubated at
15°C under dark plastic for four days (96 h). No aeration was provided.
The pH, dissolved oxygen concentration, and salinity of each test
concentration was measured at the beginning and end of the test. Floating
amphipods were counted daily, then gently tapped down with a pipet tip.
2.6.5 Statistical Treatment
2.6.5.1 Screening Surveys
Results of the amphipod bioassay in screening survey sediments were
used in a qualitative reconnaissance sense. Samples were not replicated
which allowed analysis of more stations within bays. This approach was
used to gain a more representative spatial sampling. Because there were
no replicates, certain statistical comparisons could not be made, but
samples (stations) of concern were identified if mortality exceeded the
control range. Swartz et al. (1984) indicated that for R. abronius,
survival of less than 17 of 20 individuals would indicate possible
toxicity.
2.6.5.2 Detailed Surveys
An analysis of variance (ANOVA) was performed to compare mean
survival and reburial in controls with that of treatments (sediment
sources). Before an ANOVA was performed, however, it was necessary to
apply Cochran's Test (HOMOV) (EPA/COE 1977) to determine whether the
variances of the data sets were homogeneous. The homogeneity of the
variances was determined by calculating the C-value, defined as the ratio
of the largest variance to the sum of all variances. In the situation
where variances were found to be heterogeneous, it was necessary to
transform the data. The transformation was performed by obtaining the
arcsin /Y, where X was the datum.
To determine potential differences among treatment means (when ANOVA
F-value exceeded tabulated value), the Student-Newman-Keuls (SNK) Multiple
Range Test (Sokal and Rohlf 1969) was applied. Dunnett's Test (Steel and
51
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Torrie 1960) was performed to compare the control to each treatment mean.
This procedure requires a single difference for judging the significance
of observed differences.
In reference toxicant tests, amphipod survival at the end of the 96-h
exposure period was used to calculate the LC5o for each PCP assay. The
moving average-angle_,technique was used to make these calculations
(Bennett 1952). In addition, log-linear analyses (Fienberg 1977) of the
resulting dose response curves and a chi-square analysis of a 2 x 6
contingency table were conducted to determine the presence or absence of a
batch effect.
2.7 OYSTER LARVAL BIOASSAY (PERFORMED BY MRL)
2.7.1 Strategy
In general, we followed the American Society for Testing and
Materials (ASTM) Standard Procedure, Designation E 724-80 (ASTM 1980).
The technique had to be modified, however, to perform sediment testing.
These modifications pertained mainly to the method of exposing the larvae
to the potentially toxic metals/chemicals associated with test sediments
(ASTM E 724-80 was not designed to test sediments).
2.7.2 Screening Surveys
For the 1983 oyster larval bioassay series, sediments were
collected by MRL staff as described in Section 2.1.1.2. Sediments were
stored (48 to 96 h) at 4°C until used. Ripe oysters, Crassostrea gigas,
were spawned at either MRL or at a local oyster hatchery, and the embryos
were delivered within 2 h of spawning. Sediments were weighed at 0.01,
0.10, 1.0, 10.0, and 100.0 g (each replicated three times) and placed into
glass beakers and the volume in each beaker brought up to 1 L. The
sediments were resuspended by stirring for 5 min, allowed to settle for
2 h, and then innoculated with ~30,000 developing oyster embryos.
Sand-filtered Sequim Bay seawater was used a diluent. Salinity, dissolved
oxygen, temperature, and pH of the seawater and sediment mixtures were
measured at the start and completion of each experiment, and at an interim
52
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point in some cases. Sediments assayed were collected from two stations
at the Founnile Rock - Elliott Bay dump site vicinity, two stations in
Sinclair Inlet, and one station each in Port Gardner - Everett Harbor and
Sequim Bay (see Appendix A for detailed station descriptions). Four
control replicates were run with each sediment sample assayed. Controls
consisted of sand-filtered Sequim Bay seawater without sediment. Sequim
Bay sediment from Station 14 was used in a sediment control. Station 14
is classified as sandy-silt consisting of 58.2% fines (the sum of silt and
clay fractions), and ranked as the cleanest station in Sequim Bay.
Experiments were terminated between 46 and 50 h after introduction of
the oyster larvae. The liquid portion of each experimental beaker was
then siphoned into a flask. From this flask, two 10-mL samples for
scoring were withdrawn and preserved with an equal volume of buffered 8%
formalin. The remaining suspended oyster larvae were screened and
preserved similarly. The pH of the preserved specimens was 7.5. Samples
were scored in a 10-mL counting chamber at 40x and lOOx magnification.
Samples were sheathed with a blind-coded label and not specifically
identified until the counts had been completed. Samples were enumerated
for percent normal and percent abnormal larvae. Normal larvae were those
that were fully shelled. It should be noted that some of those called
normal were slightly bean shaped and undersized, but still fit Woelke's
normal classification (Woelke 1960; 1972).
2.7.3 Detailed Surveys
The 1984 oyster larval bioassays were conducted following the same
general procedures as outlined in the preceding section. In most cases,
however, sediments were collected and processed by EPA Region 10
Laboratory staff, and delivered the following day to the MRL for bioassay.
The only exception was the Sequim Bay sediments, which were collected by
MRL staff. Also, a single dilution of sediment (100.0 g/L) was tested
instead of a range of dilutions.
Sediments assayed were collected from eight stations in each urban
bay and four stations in each baseline bay (see Appendix A for detailed
station descriptions).
53
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2.7.4 Statistical Treatment
2.7.4.1 Screening Surveys
The objective of the oyster larval bioassays in sediments from the
screening surveys was to calibrate our methods and, in particular, test
various modifications of ASTM E 724-80 (ASTM 1980) to adapt the technique
to sediment testing. Therefore, rigorous statistical comparisons were not
conducted, nor were they intended.
2.7.4.2 Detailed Surveys
An analysis of variance (ANOVA) was performed to compare the
proportion of abnormal larvae in controls with the proportion of abnormal
larvae in treatments (sediment sources). Before performing the ANOVA,
however, all data were transformed by obtaining the arcsin /X, where X was
the proportion of abnormal larvae.
Dunnett's Test (Steel and Torrie 1960) was performed to compare the
control to each treatment mean. This procedure requires a single
difference for judging the significance of observed differences.
2.8 GENERAL STATISTICAL PROCEDURES (PERFORMED BY MRL)
2.8.1 Discriminant Analysis
Stepwise discriminant analysis was used to build linear
classification functions to discriminate among samples from bays using
both physical and chemical data from the 1984 detailed survey.
The linear discrimination functions are built by examining each
variable in the observation vector and comparing "between bay" variation
to "within bay" variation. The variable that discriminates the most is
assigned to the classification function and the variance associated with
that variable is removed. The remaining variables are then examined again
for the same purpose. When a group of variables is highly correlated, and
one of the group is chosen to be included in the function, the
discriminating ability of the rest of the group is correspondingly lower.
54
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Thus, a rather large vector of observations may be reduced to relatively
few classification functions based on a reduced number of variables. The
process is described in more detail by Morrison (1976).
The technique has two limitations from the regulatory standpoint.
One, the discriminant function may turn out to be data-set dependent, and
so should be verified with independent data. Two, the variables chosen
are those which discriminate among all bays. Thus, variables that are
uniform across a number of bays (e.g., PCB-1254 in the baseline bays), are
often not powerful discriminators.
The computer program P7M from the Biomedical Computer Program
P-Series developed at the Health Sciences Computing Facility, University
of California at Los Angeles was used for the analysis. The Health
Sciences Computing Facility is sponsored by the National Insitute of
Health Special Research Resources Grant RR-3. The 1982 revision of the
program is available on the VAX® 11-780 computer, located at PNL,
Richland, Washington, which was used for the computation.
2.8.2 Factor Analysis
Factor analysis was also performed on the 1984 detailed survey
physical and chemical data. Factor analysis presupposes that a few
driving forces are behind the values of a number of variables. By
discerning these underlying factors, one may well be able to obtain a less
complicated picture of the data structure.
The final data matrix for the 1984 detailed survey (48 stations)
included the following variables measured by the MRL: sediment grain-size
(average phi); percent water; percent volatiles; concentrations of silver,
mercury, lead, IR, and total organic carbon. The data generated by EPA
included concentrations of PCB-1254, aromatic hydrocarbons, arsenic,
copper, and zinc.
® VAX is a registered trademark of the Digital Equipment Corporation,
Maynard, Massachusetts.
55
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A factor analysis performed on the matrix used the covariance
structure of the data to reduce the matrix to a smaller number of
•
so-called common factors. In the words of Morrison (1976), "Under the
factor model, each response variate will be represented as a linear
function of a small number of unobservable common-factor variates and a
single latent specific variate. The common factors generate the
covariances among the observable responses, while the specific terms
contribute only to the variances of their particular responses." It may
thereby be possible to obtain a less complicated picture of the data
structure that may enhance understanding of the occurrence of the
variables.
The computer program P4M from the Biomedical Computer Program
P-Series (University of California at Los Angeles) was used for the
analysis.
2.8.3 Correlation Analysis
Correlation analyses were performed on the sediment data to determine
the relationships among physical, chemical, and biological variables. The
resulting matrix contains 300 pairwise correlations. We recognize that
with this many correlations, the chance of a spurious correlation existing
is greater than the a = 0.05 level we reported. We have, nonetheless,
reported pairwise a levels simply to assist in making judgments about the
relative strengths of the relationships.
The final data matrix presented in Appendix E includes 25 variables
derived from the 1984 detailed survey (48 stations). These are BAY,
STATION, AGMRL (silver analyzed by MRL), HGMRL (mercury analyzed by
MRL), PBMRL (lead analyzed by MRL), ASEPA (arsenic analyzed by EPA),
CUEPA (copper analyzed by EPA), ZNEPA (zinc analyzed by EPA), DEPTH, PCH20
(percent water), GRAVEL (percent gravel), SAND (percent sand), SILT
(percent silt), CLAY (percent clay), FINES (percent fines), PCB, AH, IRMRL
(IR analyzed by MRL), PCVOL (percent volatiles), TOC, AVGPHI (average
Phi), NEMATOD (nematodes), CAPITELL (Capitella capitata). AMPHIPOD
(amphipod survival), and OYSLAR (percent abnormal oyster larvae).
56
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Computer program BMD PSD, Missing Value Correlation was used for the
analyses. The 1982 revision of the program is available on the PNL
VAX* 11-780 VAX computer, which was used for the computation.
2.8.4 Tolerance Intervals
Tolerance intervals were used to develop summary statistics useful in
determining which of the eight bays currently show signs of degraded
sediment quality (see Section 2.8.5). Tolerance intervals are used when
it is desirable to know whether a single observation falls outside the
bounds of some sample distribution. Because one is not examining
distributions of means, tolerance intervals are much wider than confidence
intervals. Tolerance intervals are established by taking the sample mean,
plus or minus a tolerance factor, times the sample standard deviation.
Tolerance factors for the normal distribution are available from the CRC
Handbook of Tables for Probability and Statistics (Beyer 1976). To attain
a tolerance factor from the table, three items need to be considered:
1. n, the number of observations in the sample distribution
2. P, the percentage of the sample distribution to include
3. a, the probability that a given observation will fall in the given
percentage of the sample distribution.
Accordingly, tolerance intervals are usually presented as "the probability
is a that P percent of the observations of the given sample distribution
fall within the tolerance interval."
2.8.5 Summary Statistics
Summary statistics presented in Section 3.7 were developed as an aid
in determining which of the eight bays currently showed signs of degraded
sediment quality. Important chemical, biological, and physical variables
as well as their critical values were selected to make this
discrimination.
® Registered trademark
57
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Six metals, (Ag, Hg, Pb, As, Cu, and Zn), were selected as variables
representing metals contamination. It was assume.d that all six metals
were of equal importance as indicators of environmental contamination.
The metals concentrations were first standardized to the same scale by
dividing each by the largest concentration of that metal found in the
sample. , The resulting scores were added and the sum divided by six to
ensure that the summary metals score was on a scale of zero to one.
A similar approach was taken with a second set of variables used to
represent levels of organic pollution. Again, it was assumed that each
organic variable was of equal importance as an indicator of environmental
contamination. The variables selected were PCB, AH, and IR. Each score
was standardized to the same scale by dividing by the largest. The
resulting numbers were added and the sum divided by three to ensure that
the summary score was on a scale of zero to one.
The next step was to calculate a critical value for the metals and/or
organics, which if exceeded would signify degraded sediment quality.
Scores from the baseline bays were assumed to be from areas of low metals
and/or organic contamination. The means and standard deviations for these
16 scores and a tolerance interval for each distribution were then
calculated using the methods described in Section 2.8.4
The three biological variables selected were amphipod mortality,
oyster larvae abnormality, and the number of capitellids at each station.
The critical value for amphipod mortality, as established in Section 3.5
was approximately 0.20. This was the level used to identify a station
with higher than expected mortality.
In the case of abnormal oyster larvae, an across-the-board critical
value could not be established. The oyster larval bioassays were not
constant in the abnormality observed in the control conditions. A level
not significantly different from control in one test may well be
significant in another. For this reason, those stations found in
Section 3.6 as showing a greater percentage of abnormal larvae when
compared to control stations' were noted as indicators of degraded sediment
quality.
58
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The number of capitell ids in the benthic infauna sampling were
included as a biological variables because these organisms are classified
as organic enrichment opportunists. No attempt was made to develop a
quantitatively defensible critical value for this variable; however,
finding a quantity of these animals in an infauna sample could indicate a
shift away from the uncontaminated environment. It was assumed that a
finding of six or more of these animals at a station indicated a condition
different from the uncontaminated or baseline condition. This was based
on the finding of no more than one capitellid at any baseline bay station,
and the finding of six or more capitellids at seven urban bay stations.
The physical variable selected was the proportion of fines in the
sediment. There is, of course, no critical value for this variable, but
it was included to further understand the data.
Because flatfish species were not caught in four of the eight bays
(Bellingham Bay, Samish Bay, Dabob Bay, Sequim Bay) because of seasonal
scarcity, the resulting pathology data were not used in the summary
statistical exercise.
59
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Page Intentionally Blank
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3.0 RESULTS
3.1 SEDIMENTARY CHARACTERISTICS (PERFORMED BY MRL)
3.1.1 Screening Surveys
During the summer of 1983, surface sediment samples were collected
at 181 stations in the eight bays (Section 2.1). These sediments were
analyzed for physical and chemical characteristics including grain size,
percent water, percent loss on ignition (percent volatile solids), Ag, Hg,
Pb, and IR (oil and grease). These "range-finding" data and the amphipod
toxicity data were used to select 48 stations, which were resampled in
1984 for additional chemical analyses and biological studies. The results
of the 1983 range-finding data are included in Table 14 and in Appendix B.
3.1.1.1 Grain Size
A variety of sediment types were sampled in each urban bay. The most
common types of sediments were sands and silts. Gravel was usually a
minor component with only four stations containing more than 2Q% gravel.
Sediments collected from greater depths usually contained greater amounts
of clay than sediments collected from shallower waters.
The sediment sampling stations in the baseline bays were selected so
that the sediment types and water depth were similar to those sampled in
the urban bays. The sediment grain size charts prepared by Roberts1 were
used to select sampling stations. The types of sediments collected in
baseline bays were almost identical to those in urban bays. Most of these
sediments were sands or muds and a few stations contained gravel. The
finest grain sediments were from the deeper stations in Case Inlet.
3.1.1.2 Water Content
Water content of sediments collected in urban bays ranged from 15%
to 71%. The lower values were from sandy sediments, and the higher
values were from sediments with high silt and clay contents. The water
1 Personal communication 1979. Original charts at the Department of
Oceanography, University of Washington, Seattle.
61
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TABLE 14. Mean Sediment Characteristics and Contaminant Levels for
48 Stations Sampled During Screening (1<
Surveys (1984) Determined by MRL
Water Volatiles Gravel Sand Silt
Bay
Samish
Dabob
Sequim
Case
Bellingham
Fourmile Rock-
Elliott Bay1
Port Gardner-
Everett Harbor
Sinclair
Overall
Mean
Year
83
84
83
84
83
84
83
84
83
84
83
84
83
84
83
84
83
84
45
55
54
47
53
63
65
65
54
68
49
49
63
62
55
60
55
58
5.0
6.0
6.0
8.0
6.0
10.0
7.0
10.0
7.0
11.0
6.0
8.0
19.0
20.0
8.0
10.0
8.4
10.9
Percent
0.0
0.3
0.5
1.3
0.2
0.2
0.1
2.8
1.4
1.8
1.2
1.1
7.2
4.6
0.7
1.8
1.8
1.9
30
16
27
53
22
22
20
25
13
11
41
46
38
32
18
24
26
29
40
59
45
25
53
44
59
38
51
59
39
31
40
44
54
45
47
44
)83) and Detailed
Clay Ag Hg
30
25
28
21
25
33
21
34
34
28
18
22
16
19
27
29
25
26
0.07
0.11
0.11
0.09
0.15
0.20
0.26
0.33
0.26
0.25
0.60
0.55
0.36
0.30
1.73
2.19
0.54
0.61
0.09
0.10
0.08
0.06
0.08
0.08
0.14
0.13
1.20
1.00
1.70
2.80
0.28
0.24
2.40
2.10
1.00
1.10
Pb
13
15
14
10
12
13
22
26
28
24
165
134
50
40
184
170
76
67
IR
141
92
22
45
48
89
139
91
1052
925
470
304
4687
1852
1350
1219
1289
743
i Dump site vicinity.
-------
content in baseline bays ranged from as low as 19% in sandy sediments to
75% in the finest grained sediments of Case Inlet. Of the 20 stations
sampled in Case Inlet, 15 had water contents greater than 65%. In the
other bays, no more than five stations per bay showed water contents
greater than 65%.
3.1.1.3 Volatile Solids
The volatile solids of urban bays ranged from 1% to 27%; however,
most of the values were in the range of 2% to 8%. Generally, volatile
solids are organic matter such as wood, bark, and marine detritus. The
volatile solids of baseline bays ranged from 0.5% to 8% with the exception
of one station in Case Inlet which showed a value of 11%. High volatile
solids are usually associated with high percent water and low percent
sand.
3.1.1.4 Infrared Spectrophotometer
The infrared spectrophotometer (IR) analysis measures the absorption
of infrared light due to the carbon-hydrogen bond in organic matter
extracted from sediment. The analysis is used as a gross measurement of
oil and grease contamination. The IR spectrophotometer was calibrated
with Prudhoe Bay crude oil, so the results, expressed in ppm dry weight of
sediment, are an indicator of petroleum content. Note, however, that
nonpetroleum sources of hydrocarbon material can also contribute to the
signal.
The IR results for urban bays ranged from 4 to 15,990 ppm. The lower
values were usually from either sandy sediments or sediments collected
away from the industrial waterways. The highest IR values were from
stations located near industrial activities.
The IR concentrations in the baseline bays were generally much lower
than in the urban bays. The IR range was 14 to 246 ppm in the baseline
bays with one value of 409 ppm found in Samish Bay.
63
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3.1.2 Detailed Surveys
During the detailed surveys (spring 1984), 48 stations were revisited
and resampled (see Section 2.1). These sediments were analyzed for
essentially the same physical and chemical characteristics that were
included in the screening surveys, and additionally for EPA priority
pollutants. The results of the 1984 detailed survey analyses are shown in
graphic form in Figures 11 through 14 and in tabular form in Table 14 and
in Appendix E.
3.1.2.1 Grain Size
The sediment grain size data are presented in three different
formats: (1) percent gravel, sand, silt and clay; (2) percent silt plus
clay; and (3) mean phi1 for each bay. The percentage of silt plus clay is
shown in Figure 11 for the 48 stations. The percentage of silt plus clay
ranged from approximately 15% to 98%, however, the mean for each bay was
in the range of 55% to 86% which would be classified as silty mud (50% to
90% silt and clay). The grand mean for the eight bays was 71% silt plus
clay. Mean sand content of the bays ranged from 12% to 44%. The gravel
content was usually not more than a few percent. When the mean grain size
was expressed as mean phi, the range was 4.2 to 5.9 phi units.
3.1.2.2 Water Content
The water content of the sediment averaged 58% for all 48 stations.
The mean for each of the eight bays ranged from 47% to 65%. The water
content of sediments increased with decreasing grain size.
3.1.2.3 Volatile Solids
The volatile solids content is a measure of the weight loss when dry
sediment is ashed in a muffle furnace at 550°C. Most of the volatile
1 The unit phi is the grain size expressed by the negative log to the
base 2 of the mean diameter in millimeters. Phi units increase with
decrease in grain size. Each unit of change, for example, 4.0 to 5.0,
results in a factor of 2 decrease in mean grain size, for example, 0.062
to 0.031 mm.
64
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100
I S ratl ttfSMirattllllllTa* If St^IiSi l«ttf?W»aMI I J « 4 8 ? _« i»l«fHH«l»
SAM SEQ
11. of Silt and Clay In
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SAM SEQ BEUJ«HAM
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12. of Volatiles in
65
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TS.Q-
«»»»«•» «*»«t»»-i n air 3 4 i ? HUBM t*«rirMnt>Mi
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SEQ 8EUJN6HAM
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EVERETT
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FI61JR^J4. IR Results for in
66
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material was organic matter, therefore, TOC content was closely correlated
with volatile solids. The mean volatile solids for each bay ranged from
6.5% to 19.5%. The mean for all of the bays was 10.9%. The volatile
solids contents of the 48 stations are shown in Figure 12. Most stations
were in the range of 5% to 15%. One station in Bellingham and five
stations in Port Gardner - Everett Harbor exceeded 15% because of wood and
tree bark in the sediments, undoubtedly derived from the local forest
products industries.
3.1.2.4 Total Organic Carbon
The pattern of TOC in sediments followed that of volatile solids
(Figure 13). The TOC content in sediments was generally in the range of
2% to 4%. A few stations from Bellingham Bay and Port Gardner - Everett
Harbor contained concentrations of TOC in the range of 5% to 16%.
3.1.2.5 Infrared Spectrophotometer
The results from the IR analyses clearly showed that baseline
sediments contained relatively low concentrations of hydrocarbon compounds
compared to most of the urban sediments (Figure 14). The mean IR values
for baseline bays were between 45 and 92 ppm, while those for urban bays
ranged from 304 ppm for the Fourmile Rock - Elliott Bay dump site vicinity
to 1852 ppm for Port Gardner - Everett Harbor.
3.2 CONTAMINANT LEVELS (PERFORMED BY BOTH EPA AND MRL)
3.2.1 Screening Surveys
The 181 sediment samples collected in 1983 were analyzed by the MRL
for Ag, Hg, and Pb. Results are presented in Table 14 and in Appendix B.
These three heavy metals were selected for the screening survey because
they are usually the metals that show the greatest enrichment in Puget
Sound sediments compared to baseline sediments. Silver enters Puget Sound
in sewage effluents (Romberg et al. 1984); lead enters Puget Sound from
many sources including sewage outfalls, rivers, industrial outfalls and
atmospheric fallout; mercury enters the Sound from industry, sewage, and
67
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rivers. These three metals are usually found enriched together in
sediments that are also contaminated with other metals, petroleum
hydrocarbons, and chlorinated organics.
3.2.1.1 Urban Bays
The concentration of Ag ranged from below the detection limit of
0.014 to 2.96 ppm. Most of the values were in the range of 0.1 to
1.0 ppm. The sediments with the highest concentrations were from Sinclair
Inlet and the lowest concentrations were from Bellingham Bay.
The Hg concentrations ranged from 0.006 to 4.9 ppm. Most values were
in the range of 0.1 to 0.6 ppm. Several values greater than the 1.0 ppm
were measured in Bellingham Bay, at the Fourmile Rock - Elliott Bay dump
site vicinity, and in Sinclair Inlet. Sinclair Inlet was the most
contaminated urban bay. Bellingham Bay was also highly contaminated as a
result of past industrial discharges of Hg (Bothner 1973). The Fourmile
Rock - Elliott Bay dump site vicinity revealed five sediment samples that
contained more than 1 ppm Hg. These same samples also contained the
highest Pb concentrations in the area, suggesting that past dredge spoil
disposal activities could have created several local hot spots. However,
many of the sediments from the dump site vicinity did not contain high
concentrations of metals when compared to other main basin sediments.
This wide range of concentrations indicated that the area surrounding the
Fourmile Rock - Elliott Bay dump site was not uniformly contaminated, and
several hot spots existed, presumably as a result of specific disposal
events.
The Pb concentrations ranged from 3 to 338 ppm. Sinclair Inlet was
the most contaminated, and the dump site vicinity also indicated high
sediment concentrations. Both Port Gardner - Everett Harbor and
Bellingham Bay showed Pb contamination, but showed much less than the
other two urban areas.
3.2.1.2 Baseline Bays
The majority of sediments in the baseline bays were contaminated with
Ag, Hg, and Pb, but to a much lesser degree than the urban bays. The
68
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concentrations of Ag, Hg, and Pb in fine-grained or muddy sediments
deposited in Puget Sound during the 1800s are on the order of 0.04, 0.04,
and 6 ppm, respectively (Romberg et al. 1984). Sandy sediments generally
contain lower concentrations than muds.
The concentrations of Ag in sediments from the baseline bays ranged
from less than 0.014 to 0.4 ppm. Case Inlet was the most contaminated
baseline bay, presumably from particulate transport from the central basin
(Riley et al. 1983). The sources of Ag deposited in Dabob and Sequim Bays
have not been identified. Possible sources include local sewage outfalls
and transport from the main basin.
The Hg concentrations in baseline bays ranged from 0.006 to 0.2 ppm.
Again, Case Inlet was the most contaminated. In Samish Bay, Hg levels
were generally higher than Ag. This was not the case for the other
baseline bays or for the urban bays except Bellingham. The ratio of Ag to
Hg in Case Inlet and Sequim Bay was approximately 2:1, similar to the
ratio in the main basin of Puget Sound. In Bellingham Bay, the ratio was
1:3 because of a local Hg source. Presumably, Samish Bay is influenced by
contaminants in Bellingham, and Case Inlet is influenced by the main basin
of Puget Sound. The sources of contaminants to Dabob Bay and Sequim Bay
are not clear.
The Pb concentrations ranged from 2 to 32 ppm in the baseline bays.
Again, Case Inlet was the most contaminated because of its proximity to
major Pb sources. Atmospheric fallout could be a significant source of Pb
to all the baseline bays. As with the other metals, Pb concentrations in
sediments were related to grain sizes. Within each baseline bay, Pb
concentrations ranged approximately an order of magnitude range in Pb
concentrations. Also, Pb concentrations correlated with water content and
volatiles, and inversely with sand.
3.2.2 Detailed Surveys
The 48 sediments collected in 1984 were analyzed by the EPA
Region 10 Laboratory for priority pollutants, which included 13 metals
69
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and approximately 100 organic compounds. Battelle analyzed the same
samples for Ag, Hg, and Pb. The EPA results are presented in graphic form
in Figures 16, 17, and 19 through 25, and in tabular form in Appendix B.
The Battelle MRL data are reported in Figures 15, 18, and 19, and in
tabular form in Table 14.
3.2.2.1 Urban Bays
Metals. Urban bay sediments were generally significantly
contaminated with Ag, As, Cu, Cd, Hg, Pb, and Zn. Sinclair Inlet
sediments demonstrated the highest average concentrations of these metals.
The other six metals (Sb, Be, Cr, Ni, Se, and Tl) were found at about the
same concentration in all bays, however, a few urban sediments were
enriched in Sb, Cr, and Ni.
The concentrations of Ag generally ranged from 0.1 to 0.5 ppm except
for five stations in the Fourmile Rock - Elliott Bay dump site vicinity.
All eight Sinclair Inlet stations demonstrated concentrations greater than
0.5 ppm (Figure 15).
Urban bays were only slightly contaminated with As, although Sinclair
Inlet showed concentrations in the range of 25 to 67 ppm at three stations
(Figure 16).
Cu concentrations in all urban bays were elevated, with values as
high as 800 ppm recorded in Sinclair Inlet (Figure 17). The mean
concentration in Sinclair Inlet was 305 ppm, which was three times the
concentration in other urban bays.
All stations in Bellingham Bay, at the Fourmile Rock - Elliott Bay
dump site vicinity, and in Sinclair Inlet were elevated in Hg. Port
Gardner - Everett Harbor, however, exhibited relatively low concentrations
of Hg similar to the baseline bays (Figure 18).
The mean concentrations of Pb in all eight bays were elevated
compared to a "baseline" concentration of 6 ppm for fine-grained sediment
(Romberg et al. 1984). All Sinclair Inlet stations and several from the
Fourmile Rock - Elliott Bay dump site vicinity were found to contain more
than 98 ppm Pb (Figure 19).
70
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4.0-
i i rutt s f 9 »s i? «101 si a >? » ,s 5 »,«i
SAM SEQ BEIUNGHAM FOURMILE
SEDIMENT STATIONS
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FIGURE 15. Silver Concentrations in
60-
E
a
a
i
o
so-
40-
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20-
10-
i $ f » t )Ti*Mir«aiiniiir34srii BJBM ••nirionaMi ii4>«rneri«tr»*a
SAM SEQ FCXWM1LE EVERETT
SEDIMENT STATIONS
FIGURE 16. Arsenic Concentrations in
71
-------
1 3
® KM2 W
SAM DA808 SEQ CASE BELUNQHUM FOURMILi
SEDIMENT STATIONS
EVERETT
FIGURE17. Copper Concentrations in Sediments
8-
a
i
I1'
o
4-
2-
.5-
fl
SAM maoB SEQ CASE seujwaHAM FOURMILE EVERETT SINCLAIR
SEDIMENT STATIONS
FIGURE 18. Mercury Concentrations in Sediments
72
-------
SCO
i t tmi ir«Mir«»in«iri4 § r 11 nan tviairtoitaiii
SAM SEQ CASE BEUJNGHAM
FIGURE 19. in
woo-
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SAM SEQ
20. Zinc in
73
-------
3.5-
i - ; :iti- if«w
-------
130
SAM S6Q B6LUNGHAM SINCLAIR
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i *«a»»»BMi ia4i*rnirtMirnii»
SAM SEQ BELUNGHAM
in
75
-------
1280-
1000-
a
a
750-
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m
o
500-
250-
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SEDIMENT STATIONS
25. in
76
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Zinc concentrations in sediments from Bellingham Bay did not appear
to be significantly elevated (Figure 20). However, several stations from
the Four-mile Rock - Elliott Bay dump site vicinity, Port Gardner - Everett
Harbor and all Sinclair Inlet stations were elevated with respect to Zn.
The pattern of Cd enrichment was similar to that of Zn (Figure 21).
Cadmium concentrations in the range of 0.1 to 0.5 ppm are not considered
enriched. As expected, all sediments from Port Gardner - Everett Harbor,
Sinclair Inlet, and several from the Fourmile Rock - Elliott Bay dump site
vicinity showed some contamination with Cd.
Two other metals that also showed little enrichment were Cr and Ni
(Figures 22 and 23). One station in Sinclair Inlet contained 129 ppm Cr
compared to a range of 40 to 80 ppm for Cr in most of the bays. Nickel
concentrations were in the range of 20 to 50 ppm for all bays except
Bellingham Bay where concentrations were in the range of 80 to 120 ppm.
Possibly, these high nickel concentrations were due to the mineralogy of
the Nooksack River drainage because since there was little or no
indication of a local source of pollution.
The quality of the analytical results for Be, Tl, Sb, and Se made it
difficult to ascertain differences among bays. There was no evidence for
contamination of sediments by Be or Tl. Many of the sediments had
undetectable concentrations of these elements. The concentrations of Sb
and Se were often undetectable at 0.1 ppm, however, Sb levels of 2.0 ppm
were detected in two sediments from Sinclair Inlet. Previous analyses for
Sb in Puget Sound sediments showed that Sb was enriched in urban sediments
of Puget Sound and correlated strongly with As (Crecelius et al. 1975).
No attempt was made in the present study to normalize the metals data
with sediment grain size. Most of those stations selected for sampling
contained fine grained sediments. We also chose to apply correlation
analyses to reveal relationships among physical, chemical, and biological
data. These results are presented in Section 3.2.3.2 and discussed in
Section 4.1.1.
Organics. The only organic compounds that were consistently
quantified in the urban bay sediments were the aromatic hydrocarbons and
77
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PCB-1254. Other compounds occasionally quantified in urban bay sediments
included phthalates and PCB-1260. For the purpose of this report, the
aromatic hydrocarbon (AH) data were presented as total aromatic
hydrocarbons for each sediment sample. Total AHs were calculated by
summing each of the individual aromatic compounds that were detected.
Usually between 3 to 10 AH compounds were detected in each urban bay
sediment.
The concentrations of total AH in sediments ranged from below the
detection limit of 60 ppb to a high of 37,000 ppb. Only three sediments
from baseline bays contained detectable AH concentrations (68 to 334 ppb).
Almost all urban bay sediments contained detectable concentrations of AHs,
but the concentrations varied greatly within each bay. Concentrations
greater than 10,000 ppb were present in one or more samples from the
Fourmile Rock - Elliott Bay dump site vicinity, Port Gardner - Everett
Harbor, and from Sinclair Inlet (Figure 24).
The concentration of PCB-1254 varied from less than 20 ppb in the
baseline bays to a mean of 579 ppb in Sinclair Inlet (Figure 25).
Bellingham Bay contained the lowest concentrations with a mean of 42 ppb.
The other urban bays each contained one or more stations with
concentrations greater than 500 ppb.
3.2.2.2 Baseline Bays
Metals. In general, baseline bay sediments contained significantly
less metals. The concentrations of Ag ranged from 0.1 to 0.5 ppm except
for one station in Case Inlet (Figure 15). The concentrations of As in
baseline bays ranged from 1.9 to 8.5 ppm (Figure 16). These are typical
concentrations for As in marine sediments (Crecelius et al. 1975). Copper
concentrations in baseline bays (Figure 17) were in the range of 28 to
74 ppm, which suggests only a slight increase above the 23 ppm measured in
sediments deposited in the 1800s (Romberg et al. 1984). Mercury
concentrations were relatively low in all baseline bays (Figure 18).
However, the mean concentrations for lead in all baseline bays (Figure 19)
were higher than a background concentration of 6 ppm for fine-grained
78
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sediment (Romberg et al. 1984). Neither zinc nor cadmium concentrations
appeared to be elevated in any baseline bay (Figures 21 and 22).
3.2.2.2.2 Organics. Of the baseline bays, only Samish Bay contained
detectable concentrations of phthalates and AHs. (Figure 24).
3.2.3 Statistical Treatment
3.2.3.1 Discriminant Analyses
Results of discriminant analyses are graphically presented in
Figures 26 and 27. The variables which produced the greatest amount of
separation (discrimination) were found to be Ag, IR, and As. The
locations of each bay or station in the respective graphs are plotted in
two-dimensional space by multiplying the concentrations of Ag, IR, and As
for that bay or station by the coefficients shown in Table 15.
Both plots seem to show that the baseline bays do not discriminate
from each other; they are all closely clustered. However, some
discrimination is observed among the urban bays, and particularly for
Sinclair Inlet. Silver is the variable in Sinclair Inlet that makes this
bay so much different than the other bays. Infrared (IR) response is the
variable that tends to make Port Gardner - Everett Harbor different.
TABLE 15. Coefficients for Canonical Variables Resulting from
Discriminant Analyses Performed on Physical and
Chemical Characteristics of Sediments Collected at
48 Stations
Coefficients for Canonical Variables
Variable X?
Ag -2.76219 0.45734 0.80779
IR -0.00014 -0.00129 0.00017
As -0.05677 0.00009 -0.09816
79
-------
312
-1
2 -2
-3
-4
-7
-5
-3
-1
-4 -2 0
CANONICAL VARIABLE 1
FIGURE 26. Discriminant Analysis of 8 Bays Using the Canonical
Variables Ag, IR, and As, where 1 is Samish Bay;
2 is Dabob Bay; 3 is Sequim Bay; 4 1s Case Inlet;
5 is Bellingham Bay; 6 is Fournrile Rock - Elliott
Bay Dump Site Vicinity; 7 is Port Gardner - Everett
Harbor; and 8 is Sinclair Inlet
80
-------
FF 0
F 6
C
A
N
0
N
I
C
A
L
V
A
R
I
A
B
L
E
-1
-2
-3
D4D
G *3*1A2 B
G
E
E EG
E E
E5
E E
7 G
-4
-S
-7
-8
FIGURE 27.
-5 -3 -1
-4 -2
CANONICAL VARIABLE 1
Discriminant Analysis of 48 Stations from 8 Bays Using
the Canonical Variables Ag, IR, and As, where A are
Stations from Samish Bay; B are Stations from Dabob Bay;
C are Stations from Sequim Bay; D are Stations from Case
Inlet; E are Stations from Bellingham Bay; F are Stations
from Fourmile Rock - Elliott Bay Dump Site Vicinity; G
are Stations from Port Gardner - Everett Harbor; and H
are Stations from Sinclair Inlet. Stations Overlapping
are Plotted as an Asterisk (*).
81
-------
3.2.3.2 Correlation and Factor Analyses
As an aid to the interpretation of the relationships among
contaminants and physical properties of the sediments, a correlation
coefficient matrix was constructed and R-mode factor analysis applied to
the 48 sediments collected in 1984. Data on 13 variables from both urban
and baseline bays were included in the factor analysis. These variables
included the six metals (Ag, Hg, Pb, As, Cu, and Zn), PCB-1254, AH, IR,
TOC, volatile solids (PCVOL), grain size (in phi units) and percent water
(PCH20). Correlation coefficients are shown in Table E.2.
Three major relationships were expressed by the correlation
coefficients: 1) the strong correlation between grain size and percent
water, 2) the strong correlations among many metals, and 3) the strong
correlations among organic contaminants, IR and TOC.
The sorted rotated factor loadings for the 13 variables in 48
sediments showed that 83% of the variance in these data was explained by
four factors (Table 16). Factor loadings of less than 0.250 have been
replaced by zero. Factor 1 was characterized by high loadings for TOC,
PCVOL, IR, AHs and smaller loadings of Zn, PCB-1254, and PCH20. This
factor reflected the association of these contaminants in sediments. The
positive correlation coefficients among such contaminants indicated that
these contaminants were bound together in contaminated sediments.
Factor 1 explained 39% of the variance in the data. Factor 2, which
explained 22% of the variance in the data, represented primarily an
organic matter factor characterized by high loadings of As, Cu, Ag, Zn,
and PCB. Factor 3 accounted for 13% of the variance and was associated
with high loadings of Pb and Hg. These heavy metals were highly
correlated, apparently because several sediments at the Fourmile Rock -
Elliott Bay dump site vicinity were highly enriched in Pb and Hg. The
fourth factor, which contributed only 9% of the variance, represented a
grain-size factor reflecting both grain size and water content.
82
-------
TABLE 16. Sorted Rotated Factor Loadings for 13
Sediment Characteristics and Contaminants
for 48 Stations
Factor 1
Factor 2
Factor 3
Factor 4
ASEPA
CUEPA
AGMRL
ZNEPA
PCB-1254
TOC
PCVOL
IRMRL
AH
HGMRL
PBMRL
AVGPHI
PCH20
0.000
0.000
0.000
0.456
0.339
0.960
0.935
0.842
0.647
0.000
0.000
0.000
0.470
0.903
0.893
0.697
0.623
0.607
0.000
0.000
0.000
0.252
0.000
0.392
0.000
0.000
0.000
0.000
0.383
0.398
0.269
0.000
0.000
0.000
0.478
0.938
0.878
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.948
0.832
83
-------
3.3 BENTHIC INFAUNA (PERFORMED BY MRL)
3.3.1 Bathymetry and Sediment Types
To understand the results of the benthic infaunal survey, it is
helpful to understand how the general physical characteristics of the
eight bays varied. Because the eight bays differed in depth and sediment
type, the stations drawn from the bays did not all derive from the same
benthic habitat.
Table 17 shows how sediment types, depth, and other physical-chemical
characteristics varied among the bays. Based on the means for these
physical characteristics, the eight bays were clustered into four groups
(Figures 28, 29, and 30). First, the Four-mile Rock - Elliott Bay dump
site vicinity and Dabob Bay were distinctly deeper (both over 100 m) and
had sediments with higher sand content and lower TOC than those of all the
rest. Second, Samish and Bellingham bays were the most shallow and had
the highest mud content of the eight. Bellingham Bay had.the second
highest average TOC level. Third, Case Inlet, Sinclair Inlet, and Sequim
Bay clustered together with intermediate mean values for depth,
percentages of silt and clay and TOC. These three bays possessed shallow
to moderate depths and sediment types of both sandy-silt and silty-mud.
Fourth, Port Gardner - Everett Harbor was distinct from the other bays in
its considerably higher mean TOC level. Among the bays of shallow to
moderate depth, Port Gardner - Everett Harbor contained the least number
of stations with silty-mud. Figure 30 summarizes the major differences of
depth and sediment characteristics among the eight bays.
3.3.2 Richness and Abundance
Bay means for the number of taxa per station (richness) and the
number of individuals per station (abundance) appear in Table 18. For
several reasons, the values for number of taxa reported here should be
considered a minimum. First, the sampling technique did not obtain
sufficient material from sediments with high clay content to fully
characterize the typically sparse infauna of such sediments. Second, the
sampling techniques could have undersampled the small crustacean component
84
-------
TABLE 17. Summary of the Distribution of Sediment
Bay
Sami sh
Dabob
Sequim
Case
Baseline
Bellingham
Fourmi 1 e Rock-
El liott Bay1
Port Gardner-
Everett Harbor
Sinclair
Urban
Types, Depth, and Other Characteristics
of the Eight Bays (48 Stations)
Number of Stations Silt
by Sediment Type Depth (m) and Clay
Sand
0
1
0
0
1
0
1
0
0
1
Silty
Sand
0
2
0
1
3
0
1
1
1
3
Sandy
Silt
0
0
2
1
3
2
5
6
4
17
Silty
Mud
4
1
2
2
9
6
1
1
3
11
Cl ayey
Mud
0
0
0
0
0
0
0
0
0
0
Mean
19
102
23
29
43
10
166
25
14
54
Range
10-30
88-113
19-26
21-40
10-113
6-17
121-189
10-98
7-18
7-189
Mean {%)
84
46
77
72
87
53
63
74
TOC (<
1.5
1.8
2.2
2.1
4.0
2.0
9.6
2.7
Dump site vicinity.
85
-------
* Four-mile Rock-Elliott Bay
160+
120+
* Dabob
80+
oo W+
* Case
* Port Gardner- * Sequim
Everett Harbor * Sinclair * Samish
* Bellingham
«.0 *f9.0 56.0 63.0 70.0 77.0 8^.0
% Silt and Clay in Sediment
FIGURE 28. Mean Depth Versus Mean Percent Silt and Clay by Bay (48
Stations). The Fourmile Rock - Elliott Bay stations were
in and around the dump site.
-------
10.0+
7.5+
5.0+
* Port Gardner-Everett Harbor
*Bel1ingham
CO
2.5+
* Dabob * Fourmlle Rock-
Elliott Bay
* Sinclair
* Case * Sequim
* Samish
0.0+
M.O W.O 56.0 63.0 70.0 77.0 8*.0
Mean Percent Silt and Clay
FIGURE 29. Mean TOC Versus Mean Percent Silt, and Clay (48
Stations). The Fourmile Rock - Elliott Bay
stations were in and around the dump site.
-------
High
TOC
00
00
* Port Gardner - Everett Harbor
* Bellingham Bay
* Fourmile Rock - Elliott Bay
* Dabob Bay
* Case Inlet
Sequim Bay
* Sinclair Inlet
* Samish Bay
Low
TOC
Deep
and
Sandy
Shallow
and
Muddy
FIGURE 30. Differences Among Bays According to Depth, Sediment Type and TOC (48 Stations).
The Fourmile Rock - Elliott Bay stations were in and around the dump site.
-------
TABLE 18. Mean Values for Number of Taxa and Number
of Individuals per Station from 48 Stations
Within 8 Puget Sound Embayments
Bay
Mean per Station
Baseline
Samish Bay
Dabob Bay
Sequim Bay
Case Inlet
Number
of Taxa
11.3
11.3
13.8
7.8
Number of
Individuals
19.3
18.0
113.5
13.5
All Baseline
11.0
41.0
Urban
Bellingham Bay
Fourmile Rock - Elliott Bay1
Port Gardner - Everett Harbor
Sinclair Inlet
7.8
9.0
7.3
9.9
71.0
17.5
113.6
82.6
All Urban
9.0
71.0
Dump site vicinity.
89
-------
of the infauna. Third, for some taxonomic groups (e.g., the polychaete
family Cirratulidae), more extensive and specialized examination under a
compound microscope is required to completely distinguish genera and
species within the group. Because richness was probably underestimated,
the benthic infauna data will not support the calculation of diversity
indices.
The benthic infauna data are best viewed as supporting their intended
role: another measure for reconnaissance rather than a definitive data
set able to support sophisticated statistical analysis. These data
indicate the numerically dominant taxa and their relative abundance and
provide a qualitative picture of the relative richness of the infauna at
selected sites in the eight bays. These data likely do not represent
conditions in all similar depositional sediments in each bay.
Based on richness and abundance, the eight bays can be classified
into four or perhaps five groups (Figure 31): 1) Sequim Bay was
characterized by the highest mean number of taxa and the second highest
mean abundance; 2) Samish and Dabob Bays were characterized by the second
highest number of taxa and low numbers of individuals; 3) Sinclair Inlet
and Bellingham Bay were characterized by intermediate to low numbers of
taxa and intermediate numbers of individuals; 4) Case Inlet and the
Fourmile Rock - Elliott Bay dump site vicinity were characterized by low
numbers of taxa and numbers of individuals; 5) Port Gardner - Everett
Harbor had the lowest number of taxa and the highest value for number of
individuals.
3.3.3 General Infaunal Patterns
At least 120 taxa of benthic infaunal organisms were present at the
48 stations from the 8 bays. For comparison, a survey by Lie (1968),
including a wider range of sediment types, and in different areas of Puget
Sound, found a total of 157 species from 48 stations. Of the 120 taxa,
52% were polychaetes, 22% mollusks, 15% crustaceans, and 10% other groups.
Following Simenstad et al. (1979), 44% were classified as detritivores,
21% as carnivores, 7% as herbivores, 19% as suspension feeders, and 15% as
undetermined or other.
90
-------
120+
* Port Gardner - Everett Harbor * Sequlm
90+
* Sinclair
* Bellingham
60+
30+
UD
* Fourmile Rock- *Samish & Oabob
* Case Elliott Bay
0+
7.0 8.0 9.0 10.0 11.0 12.0 13.0
Number of Taxa per Station
FIGURE 31. Mean Number of Individuals per Station Versus
Mean Number of Taxa per Station (48 Stations).
The Fourmile Rock - Elliott Bay stations were
in and around the dump site.
-------
From all 48 stations, the total number of individuals found was 2941.
Based on abundance, polychaetes and detritivores were the dominant
taxonomic and trophic groups in all the bays except Port Gardner - Everett
Harbor. In Port Gardner - Everett Harbor, one taxon, nematodes,
contributed the majority (almost 73%) of the number of individuals per
station. Of the mean number of individuals per station, polychaetes were
19% in Port Gardner - Everett Harbor and ranged in the other bays from 47%
(Samish Bay) to 95% (Sequim Bay). Mollusks and suspension feeders
exceeded 20% of the mean number of individuals in Samish Bay, Dabob Bay,
and the Fourmile Rock - Elliott Bay dump site vicinity, the shallow muddy
bay and the two deep sandier bays, respectively. With the exceptions just
noted, groups other than polychaetes contributed little to the number of
individuals.
Over all bays, 10 taxa accounted for 85% of the total number
of individuals; 5 taxa, for 79% (Table 19).
TABLE 19. The Ten Most Abundant Taxa for All Bays
Code
009
001
006
073
024
021
020
057
011
058
No.
Stations
Species Name Present
Cirratul ids
Nematode
Capital la capitata
Axlnopsida serricata
Nephthys cornuta francisca
Lumbri nereis spp. A
Lumbri nereis spp. B
Polydora spp.
Oorvillea moniloceras
Paraprionspio pinnata
28
8
12
22
33
12
4
7
4
11
Percent
Occur
(n/48)
58
17
25
46
69
25
8
15
8
23
No.
Indiv.
1206
753
157
113
103
40
34
34
29
28
Percent
Total
(n/2941)
41.0
25.6
5.3
3.8
3.5
1.4
1.2
1.2
1.0
1.0
Cum.
Percent
41.0
66.6
71.9
75.7
79.2
80.6
81.8
83.0
84.0
85.0
92
-------
The free-living cirratulid polychaetes (Tharyx spp. plus Chaetozone spp.)
occurred at 58% of the 48 stations and accounted for 41% of the total
number of individuals. The second most abundant taxon was a nematode
found only in two bays, Port Gardner - Everett Harbor and Bellingham Bay.
The third most abundant taxon, the capitellid polychaete (Capitella
capitata), was found in four bays and achieved large numbers only in Port
Gardner - Everett Harbor in association with the nematodes. The fourth
most abundant taxon, the mollusk, Axinopsida serricata, occurred in low
numbers in all bays except Samish Bay. The fifth most abundant taxon, the
nephtyid polychaete, Nephtys cornuta fransciscana, was the most ubiquitous
species and occurred in low numbers in all bays at 69% of the 48 stations.
3.3.4 Infaunal Patterns in Each Bay
3.3.4.1 Samish Bay
• The four stations in Samish Bay were at shallow to intermediate
depths (13-30 m) and contained sediments of silty-mud with TOC levels
below 2%. Stations 7 and 20 were more than twice as deep as Stations 1
and 3.
The infauna at all stations (Table 20) were characterized by a wide
range of richness (6 to 15 taxa per station) and by low abundance (9 to 26
individuals per station). At the two shallowest stations, polychaetes
were the most abundant group, but there were no distinctly dominant
species. At the two deepest stations, the sea cucumber, Molpadia
intermedia, was the most or second-most abundant species. These latter
stations were the only ones of all 48 stations where echinoderms were an
important component of the infauna. Samish Bay also had the highest
percentage of mollusks in its faunal list (31%).
3.3.4.2 Dabob Bay
The four stations from Dabob Bay were from depths of 88 to 113 m and
held sediments with the highest sand content found in all 48 stations.
The TOC levels were generally low (<3%). The four Stations 1, 5, 7, and
93
-------
TABLE 20. Comparison of Stations in Bays by Dominant Benthlc Infauna
Bay
SM! in
Seal in
Saalih
Sulth
Dabob
Dabob
Dabob
Dabob
Sequl*
Seoul*
Sequl*
Sequl*
Caie
Caie
Caie
Cete
Balllngha*
Belllnghe*
Belllngha*
Belllngha*
Belllnghe*
Belllngha*
Balllngha*
Belllngha*
Four*ile Rock-
Elliott Bay1
F aural la Rock-
Elliott Bay1
Fouraile Rock-
Elliott Bay1
Station
01
03
07
20
01
OS
07
15
14
17
18
20
01
11
IS
17
03
04
05
07
11
12
23
24
09
10
12
No.
Taxa
11
6
13
IS
17
13
12
3
12
14
10
19
11
10
a
2
10
10
10
1
5
11
9
6
6
7
0
No.
Indlv.
19
9
23
26
34
23
12
3
96
96
153
109
17
19
16
2
66
65
87
1
102
57
14
177
12
11
0
Depth
Percent
Sand
Percent and
(•) Cley Crave!
13
10
24
30
113
88
98
110
24
26
25
19
40
21
28
24
6
12
12
10
7
6
17
11
173
18*
189
20
26
27
26
8 ,
10
24
40
29
35
34
35
35
18
38
47
17
25
25
28
37
28
28
32
24
27
38
19
19
IS
12
BO
76
51
10
36
17
21
16
11
62
17
22
33
16
4
a
2
36
5
3
44
31
11
Percent
TOC
1.32
1.89
1.32
1.39
0.83
1.39
1.14
2.65
2.09
2.3
2.28
.2.23
2.01
1.36
2.42
2.69
12.15
4.83
2.34
3.15
2.07
3.69
2.01
2.09
1.77
2.35
2.14
Hott Abundant Spec let
Nephtyt cornuta £.
Prlonotplo clrrlfera
Nephtyt cornuta f.
Holpadla Interwdla
Axlnoptlda terrlcata
Axlnoptlda terrlcata
—
--
Clrratullda
Clrratullda
Clrratulldi
Clrratulldt
Arlcldea ip.
Arlcldea tp.
Arlcldea ip.
--
Oorvlllea *onllarl|
Hwatode
Clrratulldi
--
Clrratulldi
Clrretulldi
Clrratulldi
Clrratulldt
Axlnoptlda terrlcata
Axlnoptlda terrlcat*
—
No.
Indlv.
4
3
5
8
a
5
60
65
131
72
3
6
4
26
39
68
95
41
5
151
4
3
Percent
Total
Indlv.
21
33
22
31
24
22
63
68
86
66
18
32
25
39
60
78
93
72
36
85
33
27
Second Hoit Abundant
Sternaptlt fottor
A*phlpollt tp.
Holpadla Intermedia
Aclla entrant! i
Axlothella rubrlclncta
Axlothelle rubrlclncta
—
--
Lunbrlnerelt tpp. B
Luebrlnerett tpp. B
Paraprlonotplo pinnate
Luanrlnerelt tpp. B
Slaaabrl battl
Axlothella rubflclncta
Haplotcoloplot elongatut
--
Clrratulldi
Capital la capitate
Naphtyt cornuta t.
—
Haco*a ip.
Eteone paclflca
Clyclnde anelgera
Eteone paclflca
Prlonotplo cerrlfere
Nephtyt cornuta f.
..
No.
Indlv.
4
2
4
4
3
3
13
7
a
10
3
2
3
22
9
a
3
4
2
9
3
2
Percent
Total
Indlv.
21
22
17
IS
9
13
14
7
5
9
18
11
19
33
14
9
3
7
14
5
25
18
Ouap tlte vicinity.
-------
TABLE 20. Comparison of Stations in Bays by Dominant Benthic Infauna (continued)
Percent
Bey
Fouralle Rock-
Elliott Bey1
Fouralle Rock-
Elliott Bey*
Fourxllo Rock-
Elllott Bey1
Fouralle Rock-
Elliott Bey*
Fouralle Rock-
Elliott Bey>
Port Gerdner-
Everett Herbor
to Port Gerdner-
tn Everett Herbor
Port Gerdner-
Everett Herbor
Port Gerdner-
Everett Herbor
Port Gerdner-
Everett Herbor
Port Gerdner-
Everett Herbor
Port Gerdner-
Everett Herbor
Port Gerdner-
Everett Herbor
Slnclelr
Slnclelr
Slnclelr
Slnclelr
Slnclelr
Slnclelr
Slnclelr
Slncltlr
Station
17
20
22
23
24
01
02
03
04
05
06
07
11
06
07
06
14
17
16
19
20
No.
Texe
6
11
11
12
19
5
6
5
5
4
16
11
6
a
12
a
a
9
10
14
9
No.
Indlv.
9
25
19
29
35
130
86
97
169
335
33
30
33
50
202
61
81
73
40
137
18
Depth
JsL.
171
174
179
122
137
17
17
13
14
10
12
17
98
7
6
13
IS
IS
17
16
18
Percent
Cley
21
16
26
11
7
20
21
23
20
22
12
19
18
26
25
38
33
33
32
19
29
Send
end
Crevel
44
ta
36
76
86
37
17
37
*6
33
60
32
31
13
44
10
8
21
22
51
32
Percent
toe
1.92
1.39
1.93
1.11
3.8
6.98
4.17
11
16.42
9.22
*.51
6.67
2.29
.93
.12
.87
.04
.4
.58
.33
.28
Hott Abundent Spec let
Nephtyt cornute £.
Axlnoptlde terrlcete
Axlnoptlde terrlcete
Axlnoptlde terrlcete
Kegecrenelle coluablene
NeMtode
NeMtode
NeMtode
NeMtode
NeMtode
Cepltelle cepltete
Crutteceen ,
Axlnoptlde eerrlcete
Clrretulldt
Clrretulldt
Clrretulldt
Clrretulldt
Clrretulldt
Clrretullde
Clrretulldt
Axlnoptlde terrlcete
f
No.
Indlv.
3
14
7
10
4
109
64
84
122
279
6
12
23
16
142
50
60
60
15
111
4
•ercent
totel
Indlv.
33
56
37
34
11
8*
74
87
72
63
16
to
70
72
70
62
74
62
38
81
22
Second Hott Abundent
Axlnoptlde terrlcete
MecoM ep.
Pereonlt greclllt
Nephtyt cornute £.
Nephtyt cornute £.
Cepltelle cepltete
Cepltelle cepltete
Cepltelle cepltete
Cepltelle cepltete
Cepltelle cepltete
NeMtode
MecoM ep.
HecoM tp.
Nephtye cornute £.
Polydore brechycephele
Luabrlnerelt epp. A
Nephtyt cornute f .
Nephtvt cornute £.
Nephtyt cornute 1_.
Luabrlnerelt tpp. A
Hephtyt cornute f.
No.
Indlv.
2
2
2
6
1
16
17
11
«)
50
5
5
1
S
28
4
7
1
10
9
4
Percent
Totel
Indlv.
22
a
11
21
9
12
20
11
25
IS
IS
17
9
10
14
7
9
4
25
7
22
-------
15 constituted a gradient in sediment types from sand to silty-mud.
The characteristics of the infauna in Dabob Bay (Table 20) changed
along the sand-to-mud continuum. The sandy stations had high richness,
but low to moderate abundances. Moving from stations with the most to the
least sand, the richness and abundance decreased dramatically from 17 taxa
and 34 individuals per station at Station 1, to only 3 species and
3 individuals at Station 15. Station 1 was the third highest of the 48
stations in terms of richness, while Station 15 was one of the four lowest
in terms of both infaunal richness and abundance. Of all 48 stations,
Station 1 had the second highest sand content, while Station 15 had the
second highest clay content.
At the two sandiest stations, the bivalve mollusk, Axinopsida
serricata, was the most abundant species with the polychaete, Axiothella
rubrocincta. second in abundance. The abundances at the other two
stations were too low to distinguish any dominant species.
3.3.4.3 Seguim Bay
The four stations in Sequim Bay were of moderate depth (19 to 26 m).
Two stations had sandy-silt, and two had silty-mud sediments.
Of all of the bays, Sequim Bay demonstrated the highest mean number
of taxa per station. However, the mean number of individuals in Sequim
Bay was second to the highest. The shallowest station had the highest
number of taxa and individuals (Table 20). The stations with clay
contents of 34% to 35% possessed a diverse and abundant fauna.
Species composition varied little among the four stations. The
Cirratulid polychaetes, primarily Tharyx spp., dominated all four
stations. The next most abundant species in order were Lumbrinereis spp.,
Paraprionospio pinnata, and Nephtys cornuta franciscana. Reisch (1959,
1971) listed Prionospio pinnata (now called Paraprionospio pinnata) as
characteristic of healthy bottoms.
96
-------
3.3.4.4 Case Inlet
The four stations from Case Inlet were of moderate depth (21 to 40 m)
with sediments ranging from sandy-silt to silty-mud. In contrast to other
bays, even the sandy-silt sediment had a relatively high clay content
(47%).
Case Inlet showed moderate to low numbers of taxa and consistently
low abundances (Table 20). Station 17, with the highest clay content
(47%) of the 48 stations sampled, was represented by only two species with
a total of two individuals.
Because of the low abundances, a clearly dominant species was not
evident in Case Inlet. The polychaetes, Aricidea spp. and Sigambri bassi,
were slightly more abundant than the other species at Stations 1, 11, and
15, which had lower clay content.
As determined during the screening surveys, Case Inlet had the
highest mud content of all bays. In selecting the 48 stations to revisit
during detailed surveys, our approach for choosing stations in the
baseline bays led to selection of the sandier stations in Case Inlet.
3.3.4.5 Bellingham Bay
Bellingham Bay was the shallowest bay (depth 6 to 17 m) and contained
six stations with silty-mud and two with sandy-silt sediments. Thus, in
addition to Samish Bay, Bellingham Bay exhibited one of the highest mud
contents. Although Bellingham Bay did not show stations with clay
contents above 37%, five stations did have sand and gravel contents of
less than 10%. Accordingly, Bellingham Bay sediments demonstrated the
lowest sand content of all eight bays.
In Bellingham Bay (Table 20), three kinds of stations were
distinguished: 1) those with sparse infauna, 2) those with infauna
dominated by organic enrichment opportunist species, and 3) those with
infauna similar to that of Sequim Bay. Station 7, where only one
individual was observed, was included in the first category. Station 4,
dominated by nematodes and Capitella capitata, and Station 3 with some
97
-------
nematodes and dominated by the polychaete, Dorvlllea monilaris, were
included in the second category. Stations 5, 11, 12, 23, and 24 with
Cirratulid polychaetes in high abundance, but with no nematodes or
Capitellid polychaetes, were included in the last category.
A number of polychaete species have been classified as opportunists,
because they achieved high abundances where the bottom had been disturbed
(dredging, dumping). Of such opportunists, however, only a few of the
species appeared to be associated with successions following organic
enrichment (Pearson and Rosenberg 1978). Pearson and Rosenberg (1978)
reserved the term enrichment opportunist for Capitella capitata,
Streblospio spp. and the dorvilleid polychaetes. Stations 3 and 4 showed
infauna characteristic of organic enrichment, and both stations had
elevated TOC levels. Although Station 3 had a higher TOC level, Station 4
had a greater shift to opportunistic infauna. Station 3 had high TOC, but
it also had a high sand content. At higher levels of TOC, less infaunal
change occurred at those stations with higher sand and less clay content.
At the remaining stations, the infauna were similiar to that of
Sequim Bay. The Cirratulid polychaetes dominated the stations in
Bellingham Bay with TOC levels below 4%. Moll usks were more important in
Bellingham Bay than elsewhere, and this greater importance was probably
related to shallower depths.
3.3.4.6 Fourmile Rock - Elliott Bay
The deepest stations sampled were in the Fourmile Rock - Elliott Bay
dump site vicinity, which revealed the most heterogeneity in sediment type
among its stations. The dump site vicinity also had the highest sand
contents of the urban bays, and only one station possessed silty-mud.
The general characteristics of the infauna (Table 20) varied
considerably. Station 12, with silty-mud, was the most barren of all
48 stations, revealing no observed infauna. In contrast, Station 24, with
sand, showed the highest number of taxa of the 48 stations and moderate
abundance.
98
-------
The mollusk, Axinopsida serricata, was the most abundant species
sampled, and dominated the stations with sandy-silt and silty-sand. A
large number of polychaete species occurred at the Founnile Rock - Elliott
Bay dump site vicinity, but none in high abundance. Unlike the other bays
where polychaetes dominated, a polychaete was the most abundant species at
only one station, No. 17. This polychaete was Nephtys cornuta franciscana
found as a common but not a dominant species in the other bays.
The other bay with relatively deep stations was Dabob Bay, which also
contained sediments with high sand content. At the sand and silty-sand
stations, the Fourmile Rock - Elliott Bay dump site area had a species
composition similar to Dabob Bay, but with lower richness and abundance.
In silty-mud, stations in both bays had the lowest richness and abundance
of species of the 48 stations.
3.3.4.7 Port Gardner - Everett Harbor
The eight stations in Port Gardner - Everett Harbor were of moderate
depth (17 to 60 m) and possessed sediments of sandy-silt with relatively
low clay content. What distinguished Port Gardner - Everett Harbor from
the other bays was its high TOC levels at seven out of the eight stations.
Total organic carbon ranged from 2.23% to 16.42%.
Low richness and high abundance occurred at six of the eight stations
in Port Gardner - Everett Harbor (Table 20). Five stations were dominated
by nematodes and Capitella capitata, the organic enrichment opportunist.
Station 6 had both an elevated TOC and a high sand content. Although the
nematodes and Capitella capitata did occur there, their numbers did not
reach the high levels observed elsewhere in the bay. Swartz et al. (1982)
observed that Capitella capitata did not colonize an area recovering from
dredging until finer sediment had accumulated. At Station 11 with
sandy-silt and TOC <3%, the mollusk, Axinopsida serricata, appeared as the
most abundant species, and the nematodes and Capitella capitata were not
observed.
99
-------
Crustaceans were a more important component of the infauna in this
bay than elsewhere. Except for one station with sandy-silt and low TOC,
mollusks were absent.
Port Gardner - Everett Harbor had the most stations of all eight bays
with the most obvious change in benthic infauna.
3.3.4.8 Sinclair Inlet
Sinclair Inlet was one of the more shallow bays (7 to 18 m) and
contained sediments ranging from silty-sand through sandy-silt to
silty-mud. The infauna (Table 20) were characterized by moderate richness
(8 to 14 taxa per station) and moderate to high abundance (18 to
202 individuals per station). Silty-sand supported the highest richness
and highest abundance.
As in Sequim Bay, Cirratulid polychaetes were the most abundant taxon
at most stations. This taxon generally dominated all stations that were
characterized by silty-mud, sandy-silt, or silty-sand, and achieved their
highest abundance at stations with higher sand contents (Nos. 19 and 7).
Station 20, however, was an exception in that it was dominated by
Axinopsida serricata.
The station with highest IR and metal levels revealed the least rich
and abundant fauna; whereas, the next most contaminated station was one of
the richest in species and abundance. Station 20 in sandy-silt sediment
exhibited the lowest abundance of all infauna including Cirratulids, but
contained a very high IR (3191 ppm) signal. Station 19, with a similarly
high IR (1973), but low clay and high sand content, yielded high richness
and abundance. At stations with high sand content, the effect of an
elevated contaminant on infauna may have been less than at stations with
similiar contaminant levels but high silt and clay content. High clay by
itself appeared to support an infauna of lower numbers of taxa and
individuals.
100
-------
3.3.5 Infaunal Characteristics by Sediment Type
Placing all 48 stations into three major sediment groups 1)
silty-mud, 2) sandy-silt, 3) silty-sand or sand (Table 21), helps us
understand the relationships between infauna and sediment type. The
sediment types were not evenly distributed among the bays or by depth
(Table 17 and Figure 28). For example, Samish Bay contained only
silty-mud while Dabob Bay contained sediments with higher sand contents.
In moving from one end of this continuum to the other, the percentage of
sand in the sediment ranged from as little as 3% to an observed maximum of
86%. In general, the deepest stations had higher sand contents, while the
most shallow stations had higher mud contents.
Comparing all stations within a sediment type regardless of bay
(Table 21) showed that the stations with silty-sand or sand had distinctly
different infaunal characteristics than those with the other two sediment
types. Stations with silty-sand or sand generally had a higher number of
taxa, while stations with silty-mud and sandy-silt possessed a lower
number of taxa. Of all 48 stations, the two stations with the highest
sand content also had the highest and second highest number of taxa.
Stations with silty-sand or sand had the lowest mean abundance, while
those with sandy-silt had the highest abundance. Stations located in
sediments of silty-sand or sand showed consistently low abundances except
for one station in Sinclair Inlet. This station (No. 19) was considerably
more shallow than the rest of the stations in sandy sediments.
The sediment types supported different species compositions. In
silty-mud, the species compositions in all bays except Case Inlet and
Samish Bay were similiar. The Cirratulid polychaetes, primarily Tharyx
spp., were the most frequent dominant (45% of the silty-mud stations) and
most abundant taxon. In silty-mud, nematodes were in high abundance at
only 10% of the stations. In sandy-silt, Cirratulids were not as frequent
a dominant (30% of the stations) as in silty-mud. Nematodes had the
highest abundance followed by Cirratulids. In sandy-silt, the mollusk,
Axinopsida serricata, was dominant at 30% of the stations, but it
101
-------
TABLE 21. Comparison of the Dominant Benthlc Infauna by Sediment Type
Silty Hud
Bay
SMI) >h
Samlsh
SMlih
Samlih
Oabob
Sequin
Sequin
Case
Case
_, Belllnghan
O Bellinghan
ro Bel Unshorn
Bellinoham
Belli njham
Bellinghw
Fournilo Rock-
Elliott Bay1
Port Gardner-
Everett Harbor
Sinclair
Sinclair
Sinclair
TOC >3.8
TOC <3.8
Baseline
Urban
All Bays
Station
01
03
07
20
15
17
20
01
15
0*
05
07
11
23
2*
12
02
06
08
14
N- 2
N-18
N- 9
N-11
N=20
No.
Taxa
11
6
13
15
3
H
19
11
8
10
10
1
5
9
6
0
6
8
8
8
8
8.6
11.1
6.5
8.6
No.
Indlv.
19
9
23
26
3
96
109
17
16
65
87
1
102
14
177
0
86
50
61
81
75.5
49.5
35.3
£5.8
52.1
Depth
_J=L
13
10
24
30
110
2£
19
40
28
12
12
10
7
17
11
189
17
7
13
15
15.0
32.2
33.3
28.2
30. £
Percent
Clay
20
2£
27
26
40
35
35
35
38
25
25
28
37
28
32
38
21
26
38
33
23
31.5
31.3
30.1
30.7
Percent
Sand
and
Gravel
19
19
15
12
10
17
16
11
17
16
4
8
2
5
3
11
17
13
10
8
16.5
11.1
15.1
8.8
11.7
Percent
TOC
1.32
1.89
1.32
1.39
2.65
2.3
2.23
2.01
2.42
4.83
2.34
3.15
2.07
2.01
2.09
2.14
4.17
2.93
2.87
3.04
4.5
2.23
1.95
2.88
2.46
Host Abundant Specie*
Nephtys cornuta t.
Prionosplo cirrifera
Nephtys cornuta f.
Hoi pad I a intermedia
Cirratulids
Cirratulids
Arlcidea sp.
Aricldea sp.
Nematode
Cirratulids
Cirratulids
Cirratulids
Cirratulids
"
Nenatode
Cirratulids
Cirratulids
Cirratulids
No.
Indiv.
4
3
5
8
65
72
3
4
19
68
95
S
151
64
16
SO
60
Percent
Total
Indiv.
21
33
22
31
£8
££
18
25
60
78
93
3£
85
74
72
82
74
Second Host Abundant
Sternapsii fonor
Anphlpolii sp.
Hoi pad I a Intermedia
Aclla castrensli
«
Lumbri nereis spp. 0
Lumbri nereis spp. B
Slgambrt bassl
Haploicoloplos elongatus
Capitella capitata
Nephtys cornuta f.
—
Hacoma sp.
Clycinde aralgera
Eteone paciflca
"
Capitella capitata
Nephtys cornuta (_._
Lunbrl nereis spp. A
Nephtys cornuta f.
1 Dump site vicinity.
No.
Indlv.
4
2
4
4
7
10
3
3
9
17
S
4
7
Percent
Total
Indlv.
21
22
17
15
7
. 9
18
19
14
9
3
14
S
20
10
7
9
-------
TABLE 21.
Sandy Silt
Ray
Sequiei
Sequla
Case
Bellinghar.
UelllnghM.
Fouralle Rock-
Elliott Bay1
Fournlle Rock-
Elliott Bay1
Four-mile Rock-
Elliott Day1
Fourmfle Rock-
Ellfott Day1
Four-mils Rock-
Elliott Bay1
— J Port Gardner-
C3 Everett Harbor
OJ
Port Gardner-
Everett Harbor
Port Gardner-
Everett Harbor
Port Gardner-
Everett Harbor
Port Gardner-
Everett Harbor
Port Gardner-
Everett Harbor
Sinclair
Sinclair
Sinclair
Sinclair
IOC >3.8
IOC <3.8
Baseline
Urban
All Bays
(continued)
Station
10
18
17
03
12
09
10
17
20
22
01
03
00
05
07
11
07
17
18
20
N- 6
N- 3
N-17
N-20
No.
Taxa
12
10
2
10
11
6
7
6
11
11
5
5
5
0
11
6
12
9
10
9
6.7
8.7
8
8.1
8.1
No.
Indlv.
96
153
2
66
57
12
11
9
25
19
130
97
169
335
30
33
202
73
oo
18
137.8
53.6
83.7
78
78.9
Depth
J-L
2 ">
25
2*
6
6
173
m
171
17*
179
17
13
10
10
17
98
8
15
17
18
12.8
7*. 9
24.3
62.2
56.8
Percent
Sand
Percent and
Clay
29
3*
1(7
17
28
24
27
22
18
26
20
23
20
22
20
18
25
33
32
29
20.1
28.1
36.7
23.6
25.8
Gravel
36
21
22
33
36
1*
31
**
08
38
37
37
06
33
32
31
00
21
22
32
36.3
33.6
26.3
35.8
3*.*
Percont
roc
2.09
2.28
2.69
12.15
3.69
1.77
2.35
1.92
1.39
1.93
8.98
11
16.02
9.22
6.67
2.29
2.12
2.*
2.58
3.28
10.7*
2.3*
2.35
5.35
0.90
Moit Abundant Species
Cirratuliiis
Clrratulids
—
Dorvillea nonllaris
Cirratullds
Axinopsida serrlcata
Axinopsida serrlcata
Nephtys cornuta 1±
Axinopsida serrlcata
Axinopsida serrlcata
Mematode
Neaatode
Neaatode
Neaetode
Crustacean
Axinopsida serricata
Cirratullds
Cirratullds
Cirratullds
Axinopsida serrlcata
i
No.
Indlv.
60
131
26
3
3
10
7
109
8*
122
279
12
23
102
60
15
0
"•/ w
Percent
Total
Indlv.
63
86
39
72
33
27
33
56
37
84
87
72
83
00
70
70
82
38
22
** ** • lll%* 1 1 w 1 J ff\*
Second Host Abundant
Lunbrl nereis spp. B
Paraprlonosplo pinnate
—
Clrratulids
Eteone paclfica
Prionosplo clrrlfera
Nephtys cornuta t^
Axinopsida serrlcata
Hacorna sp.
Paraonis greet 1 is
-
Capital la capitate
Cepitelle capitate
Cepltella cap! tata
Cap! tell a capitate
Hacoma sp.
Hacoma sp.
Polydora spp.
Nephtys cornuta f.
Nephtya cornuta t._
Nephtys cornuta f.
No.
Indlv.
13
8
22
3
2
2
2
2
16
11
03
50
5
3
28
3
10
It
Percent
Total
Indlv.
U
5
33
7
25
18
22
8
11
12
11
25
15
17
9
10
25
22
-------
TABLE 21. Comparison of the Dominant BentMc Infauna by Sediment Type
(continued)
Silty Sand and Sand
Bay
Do bob
Dabob
Ca»e
Fouraile Rock-
Elliott Bay1
Port Gardner-
Everett Harbor
Sinclair
Sand
Oabob
Fouraile Rock-
Elliott Bay1
IOC >J.8
TOC <1.8
Baseline
Urban
All Bay*
Station
OS
07
11
21
06
19
01
24
N* 1
N- 7
N- 4
N- 4
N- 6
No.
Taxa
11
12
10
12
16
14
17
19
16
11.9
11
1S.1
14.1
No.
Indlv.
21
12
19
29
13
117
14
IS
13
41.1
22
S8.S
40.1
Depth
(«)
80
98
21
122
12
16
111
117
12
8S.O
80.0
71.8
7S.9
Percent
Clay
10
24
18
11
12
19
8
7
12
11.9
IS
12.1
11.6
Percent
Sand
and
Gravel
76
SI
62
76
60
SI
80
86
60
68.9
67.1
68.1
67.8
Percent
TOC
1.19
2.14
1.16
1.11
4. SI
2.11
0.81
1.8
4. SI
1.8S
1.41
2.94
2.18
Mo»t Abundant Spec let
Axinopslda lerrlcata
•
Arlcldea »p.
Axinopslda fterrfcata
Capltella capltata
Clrratulldi
Axinopslda lerricata
Hegacrenella colunblana
No.
Indlv.
S
6
10
6
111
8
4
Percent
Total
Indiv.
22
32
34
18
81
24
11
Second Host Abundant
Axlothella rubriclncta
—
Axlothella rubriclncta
Nephtys cornuta f.
Nenatode
Lunbrlnerels spp.
Axlothella rubriclncta
Nephtys cornuta f.
*
No.
Indlv.
1
2
6
5
9
1
1
Percent
total
Indiv.
13
11
21
IS
7
9
9
-------
occurred in low numbers. In silty-sand or sand, the mollusk, Axinopsida
serrlcata, was the most frequent dominant (38% of the stations).
The infaunas of Case Inlet and Samish Bay did not follow this general
pattern of change with sediment type. Case Inlet stations consistently
demonstrated the lowest richness and abundance within each sediment type.
The polychaete, Aricidea spp., was the most frequent dominant in both
silty-mud and silty-sand. Of all of the stations in silty-sand, those in
Case Inlet also exhibited the lowest number of taxa. Each station in
Samish Bay revealed a different dominant species and low abundance. In
both Case Inlet and Samish Bay, the most abundant species contributed less
than 33% to the total number of individuals regardless of sediment type.
In the other seven bays, the most abundant species typically contributed
50% to 90% of the total number of individuals at a station.
In all sediment types, high TOC was associated with lower richness
and elevated abundance, and a shift in species composition to organic
enrichment opportunists. Such shifts occurred in all sediment types, but
were observed more often in sandy-silt than in silty-sand and silty-mud.
Table 22 shows how the infauna differed with sediment type without
the influence of elevated TOC. In the table those stations with TOC
levels greater than 4% (the mean TOC level of all 48 stations was 3.8%)
and those stations from Case Inlet and Samish Bay are omitted. After
partitioning-out the influence of high TOC and the distinctly different
infauna of Samish Bay and Case Inlet, a consistent pattern of variation in
infaunal characteristics with sediment type became more evident. Richness
decreased consistently over a continuum from sand through silt to mud.
Abundance was much lower in silty-sand and sand than in the other sediment
types. The more sandy stations would have demonstrated even lower
abundance except for one station from Sinclair Inlet. This station was by
far the most shallow of all stations and had a high sand content.
Silty-mud and sandy-silt stations had equivalent abundances when TOC
levels were partitioned-out.
105
-------
TABLE 22. Comparison of the Dominant Benthlc Infauna by Sediment Type (Stations with
High TOC and Stations from Case Inlet and Samish Bay omitted)
Silty Mud
Bay
Dabob
Sequia
Sequia
Bellinghaa
BelUnjhan
Belllnghaa
Belllnghaa
Belllnghaa
Fouraile Rock-
Elliott Bay1
Sinclair
Sinclair
Sinclair
Baseline
Urban
All Bays
Station
15
17
20
OS
07
11
23
24
12
06
08
14
N- 3
H- 9
H-12
No.
Tana
3
14
19
10
1
5
9
6
0
8
8
8
12
6.1
7.6
No.
Indlv.
3
96
109
87
1
102
14
177
0
50
61
81
69.3
63.7
65.1
Depth
J-L
110
26
19
12
10
7
17
11
189
7
13
15
51.7
31.2
36.3
Percent
Sand
Percent and
Clay
40
35
35
25
28
37
28
32
38
26
38
33
36.7
31.7
32.9
Gravel
10
17
16
4
8
2
5
3
11
13
10
6
14.3
7.1
8.9
Percent
TOC
2.65
2.3
2.23
2.34
3.15
2.07
2.01
2.09
2.14
2.93
2.87
3.04
2.4
2.5
2.5
Host Abundant Species
--
Clrratulldt
Clrratulldi
Clrratulldt
~
Clrratulldi
Clrratulldi
Clrratulldt
Clrratulids
Clrratulids
Clrratulids
Ho.
Indiv.
65
72
68
95
5
151
36
SO
60
Percent
Total
Indlv.
68
66
78
93
36
85
72
82
74
Second Holt Abundant
—
Lumbrinerels tpp. B
Lumbrinerelt tpp. B
Nephtyi cornuta 1_^
—
Hacoaa sp.
Clyclnde aralgera
Eteone paciflea
Nephtyi cornuta f.
Lunbrinerets sop. A
Nephtys cornuta f.
Percent
Ho. Total
Indlv. Indlv.
7
10
3
14
5
10
7
9
Dump site vicinity.
-------
TABLE 22. Comparison of the Dominant Benthic Infauna by Sediment Type (Stations with
Sandy Silt
Bay
Sequin
Sequioi
Belllnghan
Fournlle Rock-
Elliott Bay1
Fouralle Rock-
Elliott Bay1
Fouralle Rock-
Elliott Bay1
Fouralle Rock-
Elliott Bay1
Fournlle Rock-
Elliott Bay1
Port Gardner-
Everett Harbor
Sinclair
Sinclair
Sinclair
Sinclair
Baseline
Urban
All Bays
. High TOC and
Station
14
18
12
09
10
17
20
22
11
07
17
18
20
N- 2
M-11
N-13
No.
Taxa
12
10
11
6
7
6
11
11
6
12
9
10
9
11
8.9
9.2
No.
Indiv.
96
153
57
12
11
9
25
19
33
202
73
40
18
124.5
45.4
57.5
Stations
Depth
J-L
25
6
173
184
171
174
179
98
8
15
17
18
24.5
94. 8
84.0
from Case Inlet and Samish Bay
Percent
Sand
Percent and
Clay
29
34
28
24
27
22
18
26
18
25
33
32
29
31.5
25.6
26.5
Gravel
36
21
3C
44
31
44
48
38
31
44
21
22
32
28.5
35.5
34.5
Percent
TOC
2.09
2.28
3.69
1.77
2.35
1.93
1.39
1.93
2.29
2.12
2.4
2.58
3.28
2.2
2.3
2.3
Most Abundant Species
Clrratulids
Cirratulids
Clrratulids
Axlnopslda serrleata
Axlnopslda serrleata
Nephtys cornuta t._
Axlnopslda serrleata
Axinopsfda serrleata
Axlnopslda serrleata
Clrratulids
Cirratulida
Cirratulids
Axinopsida lerrlcata
No.
Indlv.
60
131
41
4
3
3
14
7
23
142
60
IS
4
Omitted) (Continued)
\
Percent
Total
Indlv.
63
86
72
33
27
33
56
37
70
70
82
38
22
Second Host Abundant
Lumbrlnerels spp. B
Peraprlonospio pinnata
Eteone pad flea
Prlonospio clrrlfera
Nephtys cornuta Jf^
Axinopsida serrleata
Macoma sp.
Paraonls gracllis
Macoma sp.
Polydora spp.
Nephtys cornuta f.
Nephtys cornuta f.
Nephtys cornuta f.
No.
Indlv.
13
8
4
3
2
2
2
2
3
28
3
10
4
Percent
Total
Indlv.
14
5
7
25
18
22
8
11
9
14
4
25
22
-------
TABLE 22. Comparison of the Dominant Benthic Infauna by Sediment Type (Stations with
High TOC and Stations from Case Inlet and Samish Bay Omitted) (Continued)
Silty Sand and Sand
Percent
Bay
Dabob
Dabob
Sand
No. No. Depth Percent and
Station Taxa Indiv. (•) Clay Gravel
05 13 23 88 10 76
07 12 12 98 2* 51
Percent No.
TOC Moat Abundant Specie* Indlv.
1.39 Axlnopslda serrlcata S
2.1*
Percent
Total
Indlv.
Second Most Abundant
22 Axiothella rubrtcincta
Percent
No. Total
Indlv. Indlv.
3 13
O
00
Fourmlle Rock-
Elliott Bay1
Sinclair
Sand
Dabob
23
19
01
12 29 122 11 76
H 137 16 19 51
17 3» 113
Sand
Fourmlle Rock- 2* 19 35 137
Elliott Bay1
.Baseline N- 3 1* 23
Urban N- 3 15 67
All Bays. N- 6 1».S 45
8 80
8G
99.7 H
91.7 12.3
95.7 13.2
69
71
70
1.11 Axlnopslda terrlcata 10 3» Nephtys cornuta 1.
2.33 Clrratulldt
111 81 Lunbrlnerel» »pp. A
0.83 Axlnopslda serrlcata 8 2» Axlothella rubrtcincta
3.8 Heaacrenella sp.
1.5
1.9
11 Mephtys cornuta f._
21
-------
Without the influence of high TOC, species composition in silty-mud
was primarily dominated by Cirratulid polychaetes, primarily Tharyx
mom'laris. In sandy-silt, Cirratulids were the most abundant taxon at
half of the stations, whereas Axinopsida serricata was the most abundant
species in the other half. Within sandy-silt, Cirratulids were dominant
at stations with the highest clay and lowest sand content, whereas
Axinopsida serricata was dominant at stations with higher sand contents.
In silty-sand and sand, Axinopsida serricata was the common dominant.
Cirratulids were the most abundant taxon at the most shallow sandy
station. From Table 22, Nephtys cornuta franciscana was commonly the
second-most abundant species found in urban bays. Its contribution to the
total number of individuals was greater in sandy-silt than in silty-sand
and sand, and smallest in silty-mud.
In summary, along the continuum from shallow muddy stations to deep
sandy stations, the species composition of those stations without high TOC
changed from one dominated heavily by Cirratulid polychaetes in silty-mud
to one dominated by Axinopsida serricata in sand. Along the same
continuum, the number of taxa increased while the number of individuals
decreased.
3.3.6 Statistical Treatment
3.3.6.1 Correlation Analyses
The correlation matrix presented in Appendix E was again used to
interpret relationships among biological indices and physical and chemical
properties of the sediments. As before, data on 25 variables from both
urban and baseline bays were included.
The station abundances of nematodes and Capitata capitella were
positively correlated with water content, grain size, and AH. The
strongest positive correlations (p <0.001) of abundance of opportunists
occurred with percent volatile solids and TOC. The
109
-------
abundances of the opportunist infauna were also strongly, but negatively,
correlated (p <0.001) with amphipod survival (i.e., opportunist abundances
were high at stations where amphipod survival was lowest).
3.4 FISH AND SHELLFISH PATHOLOGY (PERFORMED BY NOAA)
3.4.1 Fish Pathology
A total of 138 English sole (Parophrys vetulus) and 83 Dover sole
(Microstomus pacificus) were collected and examined from 10 stations. A
broad spectrum of pathologic conditions were observed in each species.
These lesions were classified as either infectious (lesions associated
with infectious agents, such as a virus, rickettsia, or a protozoan,
mesozoan, or metazoan parasite) or idiopathic (lesions having no apparent
association with infectious organisms). Because the results of previously
conducted studies (Maiins et al. 1982, 1984; McCain et al. 1982) on
diseases of wild flatfish populations in Puget Sound have indicated that
the liver, kidney, and gill are most often affected with lesions of an
idiopathic nature, the focus of the present study was on idiopathic
conditions of these organs. These lesions may be caused by chemical
contaminants, nutritional imbalances, genetic disorders, microorganisms
(not distinguishable at the light microscope level), trauma, or as yet
undefined environmental factors.
3.4.1.1 English Sole
Liver Lesions. Microscopic lesions of the liver were categorized
into eight broad classifications based on histopathologic features. These
classifications were 1) neoplastic conditions, 2) preneoplastic conditions
(hyperplasia/foci of cellular alteration), 3) specific degenerative
lesions, 4) storage disorders, 5) nonspecific degenerative lesions,
6) vascular disorders, 7) proliferative disorders, and 8) inflammation
(Maiins et al. 1982, 1984). The incidences of these liver lesions in
110
-------
English sole.are shown by area and station in Tables 23 and 24. Hepatic
neoplasms were detected only in sole from Sinclair Inlet and Case Inlet.
The incidence of hepatic neoplasms is not significantly higher than the
overall incidence of neoplasms in English sole from all areas sampled in
this study. Similarly, preneoplastic conditions were found only in
Sinclair and Case inlets, with the incidence of these lesions at Station 6
in Sinclair Inlet being significantly higher than at any of the other
stations. English sole with specific degenerative hepatic lesions were
found only in the Fourmile Rock - Elliott Bay dump site vicinity and
Sinclair Inlet; the incidence was highest at Station 23 in Elliott Bay.
Storage disorders in the liver were generally found to be widely
distributed throughout the sampling sites, with significantly lower
incidences at the baseline stations in Case Inlet and offshore Eliza
Island and significantly higher incidences at the stations in Sinclair
Inlet. The incidences of the other five types of hepatic lesions in
English sole were either generally similar at a number of sampling
stations, or the incidences were very low (Tables 23, 24).
Because the incidence of some liver lesions in English sole has been
found to increase with age, the fish from each station were classified
into five size classes that corresponded approximately to five age
categories ranging from 1+ year class to >5+ year class. Table 24 gives
the number of fish examined and the incidences of liver lesions for each
size class by groups of stations (i.e., all reference sites, all Elliott
Bay sites including West Point, and all Sinclair Inlet sites). One
instance of significantly greater lesion incidence was found among the >5
size class. Fish in this size class from the Sinclair Inlet stations were
found to have more preneoplastic conditions than fish of comparable size
from either the reference group or the Elliott Bay stations. On the other
hand, four instances of significantly lower lesion incidences in
individual size classes were found. In size class 2, significantly lower
incidences of preneoplasms were found among fish from the reference group
when compared to the Elliott Bay and Sinclair Inlet groups. Among the
111
-------
TABLE 23. Incidences (%) of Idiopathic Liver Lesions in English Sole from Stations
Sampled in Puget Sound, March 1984
Lesion Types (%)
I.D.
(NOOC *)
Case 5
(12134)
Eliza Is.
(04015)
Dabob 15
(07026)
2
Fourmile Rock-
Elliott Bay 17
(10061)
o
Fourmile Rock-
Elliott Bay 23 '
(10063)
West Pt.
(10065)
Sinclair 17
& 18
(08009 &
08006)
Sample
Size
30
28
1
5
12
2
30
Neoplasms
3.3
0.0
0.0
0.0
0.0
0.0
0.0
Preneo-
plastic
Lesions
3.3
O.O(L)
0.0
0.0
0.0
0.0
10.0
Specific
Degenera-
tive/
Nee rot ic
0.0
0.0
0.0
0.0
25.0(H)
50.0
0.0
Storage
6.7U)1
7.KL)
0.0
0.0
16.7
0.0
46.7(H)
Non~
Specific
Degenera-
tive/
Necrotic
43.3
7.1
0.0
0.0
16.7
0.0
33.3
Vascular
Disorders
0.0
0.0
0.0
0.0
0.0
0.0
3.3
Prolif-
erative
Disorders
0.0
0.0
0.0
0.0
0.0
0.0
0.0
Inflam-
mations
3.3
O.O(L)
0.0
0.0
0.0
0.0
16.7
Any
Idiopathic
Liver
Lesions
46.7
14.3(L)
0.0
O.O(L)
41.7
50.0
70.0(H)
L indicates stations classified in the low incidence group, and H Indicates stations in the high incidence
2 group. Significance level for G-statistic was p "-0.05.
Dump site vicinity.
-------
TABLE 23. Incidences (%) of Idiopathic Liver Lesions in English Sole from Stations
Sampled in Puget Sound, March 1984 (Continued) *
Las Ion Types (%)
I.D.
(NODC #)
Sinclair 6
(08010)
All Ref
Stations
(Case, Eliza,
Da bob)
All Sinclair
Stations
All Elliott
Stations
Preneo-
Sample plastic
Size Neoplasms Lesions
30 6.7 30.0(H)
59 1.7 1.7(L)
60 3.3 20.0(H)
19 0.0 0.0
Specific
Degenera-
tive/
Nee rot ic Storage
10.0 53.3(H)
O.O(L) 6.8(L)
5.0 50.0(H)
21.1(H) 10.5
Non-
specific
Degenera-
tive/ Vascular
Necrotic Disorders
46.7 6.7
25.4 0.0
40.0(H) 5.0
10.5(L) 0.0
Prolif-
erative I nf lam-
Disorders nations
3.3 20.0
0.0 1.7(L)
1.7 18.3(H)
0.0 0.0
Any
Idiopathic
Liver
Lesions
73.3(H)
30.5(H)
71.7(H)
31.6
-------
TABLE 24. Incidences (%) of Idiopathic Liver Lesions in
English Sole from Reference and Urban Areas by
Size Class '
Number examined
Reference
Elliott Bay
Sinclair Inlet
Percent neoplasms
Reference
Elliott Bay
Sinclair Inlet
Percent "preneoplasms"
Reference
Elliott Bay
Sinclair Inlet
Percent specific
degenerati on/necrosi s
Reference
Elliott Bay
Sinclair Inlet
Percent storage disorders
Reference
Elliott Bay
Sinclair Inlet
Percent nonspecific
necrosis
Reference
Elliott Bay
Sinclair Inlet
Percent vascular disorders
Reference
Elliott Bay
Sinclair Inlet
Size Class1
1
14
0
11
0.0
-
0.0
0.0
-
9.1
0.0
-
0.0
14.3
-
0.0
7.1
-
18.2
0.0
-
0.0
2
34
1
1
0.0
0.0
0.0
O.O(L)2
0.0
0.0
0.0
100.0
0.0
2.9
0.0
100.0
20.6
100.0
0.0
0.0
0.0
0.0
3
4
4
3
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
50.0
0.0
33.3
0.0
0.0
0.0
4
2
7
1
0.0
0.0
0.0
0.0
0.0
0.0
0.0
28.6
0.0
0.0
28.6
0.0
100.0
14.3
0.0
0.0
0.0
0.0
5
5
7
43
20.0
0.0
4.6
20.0
0.0
25.6(H)2
O.O(L)
14.3
7.0
20.0
0.0{L)
51.2
60.0
O.O(L)
48.8
0.0
0.0
7.0
Size class corresponds approximately to age class as follows:
Size class 1 2 3 4 5
„ Age class 1+ 2+ 3+ 4+ >5+
L indicates stations classified in tHe low incidence group, and H indicates stations
in the high incidence group. Significance level for C-statistic was p <0.05.
114
-------
TABLE 24. Incidences (%) of Idiopathic Liver Lesions in
English Sole from Reference and Urban Areas by
Size Class (Continued)
Size Class*
Percent proliterative
disorders
Reference 0.0 0.0 0.0 0.0 0.0
Elliott Bay - 0.0 0.0 0.0 0.0
Sinclair Inlet 0.0 0.0 0.0 0.0 2.3
Percent inflammations
Reference 7.1 0.0 0,0 0.0 0.0
Elliott Bay - 0.0 0.0 0.0 0.0
Sinclair Inlet 9.1 0.0 33.3 0.0 20.9
Any idiopathic liver lesion
Reference 21.4 23.5 50.0 100.0 60.0
Elliott Bay - 100.0 0.0 57.1 14.3(L)2
Sinclair Inlet 36.4 100.0 66.7 0.0 83.7
Size class corresponds approximately to age class as follows:
Size class 12345
Age class 1+ 2+ 3+ 4+ >5+
L indicates stations classified in tHo low incidence group, and H indicates stations
in the high incidence group. Significance level for G-statistic was p <0.05.
115
-------
class of 5+ year fish, significantly lower incidences of lesions were
found among fish from the Elliott Bay stations.
Kidney Lesions. Lesions were primarily observed in the excretory (as
opposed to the hematopoietic) component of the kidney. Lesions were
detected in the glomerular and tubular segment of the of the nephron, and
were categorized as 1) depositional disorders (mesangiosclerosis of the
glomeruli, and hypermembranous glomeruli), 2) necrotic lesions
(mesangiolysis of the glomeruli, tubular epithelial necrosis and tubular
epithelial vacuolization), and 3) proliferative lesions (mesangial
hypercellularity in the glomeruli, proliferation of the visceral/parietal
layers of Bowman's capsule). All three of the above categories were found
in the glomerular segment, whereas necrotic lesions were almost
exclusively found in the tubular segment. Depositional disorders were
found only in sole from Sinclair Inlet, with a higher incidence at
Station 6 than at other Sinclair stations combined (Table 25). Necrotic
and proliferative lesions were widely distributed throughout the areas
sampled, with only the incidence of necrotic lesions in sole from
Station 6 in Sinclair Inlet being higher. A higher incidence of one or
more idiopathic kidney lesions was also observed in sole from this station
and in Sinclair Inlet in general.
Gill Lesions. Three principal types of gill lesions were detected:
proliferative disorders, vascular disorders, and inflammation.
Proliferative lesions were observed in the respiratory epithelium,
filament epithelium, and mucous cells of both epithelial components.
Respiratory epithelial hyperplasia (a proliferative disorder) was often
associated with fusion of adjacent gill lamellae and/or filaments.
Proliferation of the pillar cells forming the internal structural
component of the lamellae was also found. Vascular disorders were
characterized by multiple sites of intralamellar capillary dilations,
microaneurysms or telangiectasis. These occurred in absence of
116
-------
TABLE 25. Incidences (%) of Idiopathic Kidney and Gill Lesions
in English Sole
Kidney Lesions (»)
I.D.
(NOOC *)
Case 5
(12134)
Eliza Is.
(04015)
Oabob 15
(07026)
o
Fourmile Rock-
Elliott Bay 17
(10061)
o
Fourmile Rock-
Elliott Bay 23
(10063)
West Pt.
(10065)
Sinclair 17
& 18
(08009 &
08006)
Sample
Size
30
26
1
5
12
2
30
Oesposltlonal
Disorders
O.O(L)1
0.0
0.0
0.0
0.0
0.0
6.7
One or
Prol iferatlve More
Nee rot ic
6.7
3.9
0.0
20.0
0.0
0.0
13.3
Lesions
20.0
30.0
0.0
20.0
25.0
0.0
23.3
Lesions
26.7
30.8
0.0
40.0
25.0
0.0
40.0
Sample
Size
30
26
1
4
12
2
28
Gill Lesions (%)
Prol Iferatlve Vascular
Lesions
3.3(L)
23.1
0.0
0.0
O.O(L)
0.0
42.8(H)
Disorders
6.7
19.2
0.0
25.0
16.7
0.0
10.7
Inflam-
mation
56.7
61.5
0.0
50.0
41.7
50.0
42.8
One or
More
Lesions
56.7
73.1
0.0
75.0
41.7
50.0
67.8
L indicates stations classified in the low incidence group, and H indicates stations in the high incidence group.
2 Significance level for C-statistic was p ^.OS.
Dump site vicinity.
-------
TABLE 25. Incidences (%) of Idiopathic Kidney and Gill Lesions
in English Sole (Continued)
Kidney Lesions (%)
I.D.
(NODC *)
Sinclair 6
(08010)
All Ref
Stations
(Case, Eliza,
Da bob)
All Sinclair
Inlet
All Elliott
Bay
Sample
Size
30
57
60
19
Desposltlonal
Disorders
25.9(H)
0.0
15.8(H)
0.0
One or
Proliferatlve More
Nee rot ic Lesions Lesions
25.9(H) 33.3 66.7(H)
5.3 24.6 28.1
19.3 28.1 52.6(H)
5.3 21.1 26.3
Sample
Size
30
57
58
18
Gill Lesions (%)
Prollferatlve Vascular
Lesions Disorders
53.3(H) 20.0
12.3 12.3
48.3(H) 15.5
O.O(L) 16.7
One or
Inflam- More
matlon Lesions
40.0 30.0
57.9 63.2
41.4 74.1
44.4 50.0
00
-------
hemorrhage, but the dilated capillaries often contained fibrin
accumulations indicative of a coagulation disorder or a tendency towards
thrombosis, possibly from endothelial injury. Most commonly, however, the
lesion consisted of simple capillary dilation and congestion, possibly
associated with the shock response induced in capture. Inflammation
generally consisted of multifocal sites of lymphocytic and histiocytic
infiltration in the subepithelial regions of the gill lamellae and
filaments. There was generally no accompanying necrosis, and this
condition can be interpreted as a nonspecific chronic response to injury.
Proliferative lesions of the gill were most prevalent in sole from
Sinclair Inlet and near Eliza Island - Bellingham Bay; the incidence of
these lesions was highest at stations in Sinclair Inlet (Table 24). This
condition was not detected in sole from any Elliott Bay site and was found
in only one sole from Case Inlet. Vascular disorders and inflammatory
conditions were found in sole from most of the sampling stations at
generally similar incidences.
3.4.1.2 Dover Sole
Liver Lesions. Liver abnormalities observed in Dover sole were
similar to those described for English sole, except no hepatic neoplasms
were detected in Dover sole. Preneoplastic liver lesions were detected
only in Dover sole from Station 17 in Fourmile Rock - Elliott Bay (6.7%, 2
of 30 fish) (Table 26). Specific hepatocellular degenerative lesions were
found only in sole (10%, 3 of 30 fish) from the station near West Point in
Elliott Bay. The incidences of storage disorders, nonspecific
degenerative lesions, vascular disorders, and proliferative lesions in
sole were generally similar at all of the Elliott Bay stations. The
incidence of inflammatory lesions in Dover sole was significantly higher
at Station 17 (23.3%, 7 of 30 fish), although this condition was also
found at the West Point Station (6.7%, 2 of 30 fish).
119
-------
TABLE 26. Incidences (%) of Idiopathic Liver Lesions in Dover
Sole from Elliott Bay
Lesion Incidence (%) at Each Station
Four-mi le Rock-1
Elliott Bay 17
Lesion Type 10061
N-
Neoplasms
Preneoplasms
Specific
degenerative
Storage disorders
Nonspecific
degenerative
Vascular disorders
Proliferative
Inflammation
Any lesion
30
0.0
6.7(H)2
0.0
13.3
30.0
3.3
3.3
23.3(H)
53.3
Fourmile Rock-
Elliott Bay 23
10063
20
0.0
0.0
0.0
15.0
5.0
0.0
5.0
O.O(L)
25.0
West Pt.
10065
30
0.0
0.0
10.0
10.0
13.3
6.7
0.0
6.7
33.3
Ouwami sh
Head
10064
3
0.0
0.0
0.0
0.0
33.3
0.0
0.0
0.0
33.3
All
Elliott
Bay
83
0.0
2.4
3.6
12.1
18.1
3.6
2.4
10.8
38.6
Dump site vicinity.
L indicates stations classified in the low incidence group, and H indicates stations
in the high incidence group. Significance level for G-statistic was p<0.05.
120
-------
3.4.1.2.1 Kidney Lesions. Very low incidences (3.3%) of the kidney
lesions previously described for English sole were detected in Dover sole
(Table 26). Only Dover sole from the West Point Station had detectable
kidney lesions.
Gill Lesions. Gill lesions previously described for English sole
were also observed in Dover sole. Gill necrosis and proliferation were
found only in Dover sole from the West Point Station, 3.5% (1 of 28 fish)
and 21.4% (6 of 28 fish), respectively (Table 27). The incidences of
vascular disorders and inflammatory lesions in Dover sole were generally
similar at all of the Elliott Bay Stations.
3.4.2 Shellfish Pathology
A total of 148 Cancer magister. 73 Cancer gracilis, 55 Panda!us
platyceros, and 53 Pandalopsis dispar were collected and examined from
14 stations. Histopathological conditions were placed into several
general categories, as follows.
1. vascular disorders: congestion and microaneurysms
2. inflammatory conditions: hemocytic responses, melanized lesions, and
unmelanized granulomata
3. degenerative disorders: cellular degenerative changes, nuclear
atypia, and hydropic degeneration with membrane lyis
4. proliferative conditions as reflected by increased mitotic activity
and
5. parasitic/infectious disorders.
Vascular disorders were typically confined to the gill, and
congestion of the giU filaments by hemocytes and microaneurysms (marked
dilation of gill filaments caused by hemolymph accumulation in the hemal
space) was the most common condition.
121
-------
TABLE 27. Incidences (%} of Idiopathic Kidney and Gill Lesions
in Dover Sole from Elliott Bay
Lesion Incidence (%) at Each Station
Four-mile Rock-*
Elliott Bay 17
Lesion Type 10061
Lesion Type
Kidney
N=
Deposition
Necrotic lesions
Proliferative
Inflammation
One or more
lesions
Gill
N=
Necrotic lesions
Proliferative
Vascular disorders
Inflammation
One or more
lesions
29
0.0
0.0
0.0
0.0
0.0
29
0.0
0.0
31.0
65.5
72.4
Four-mile Rock-*
Elliott Bay 23 West Pt.
10063 10065
19
0.0
0.0
0.0
0.0
0.0
20
0.0
0.0
35.0
75.0
85.0
30
0.0
3.3
0.0
0.0
3.3
28
3.6
21.4
42.9
78.6
85.7
Duwami sh
Head
10064
3
0.0
0.0
0.0
0.0
0.0
3
0.0
0.0
0.0
33.3
33.3
All
Elliott
Bay
81
0.0
3.3
0.0
0.0
3.3
80
1.3
7.5
35.0
71.3
78.8
1 Dump site vicinity.
122
-------
Inflammatory conditions included hemocytic responses, which are
thought to be analogous to the short-term inflammatory response in
vertebrates. These responses typically were represented by hemocyte
infiltration into supportive connective tissues or epithelial layers,
hyalinocytic encapsulation and/or hyalinocytic phagocytosis. Lesions that
probably represent an analog to the chronic inflammatory response in
vertebrates were melanized lesions and unmelanized granulomata, which are
general, nonspecific responses to injury or parasitic/infectious agents
(Bang 1983; Fontaine and Lightner 1975; Sparks 1980).
Lesions within the degenerative category were represented by a
broad spectrum of conditions characteristic of degenerative or dying
cells. These lesions included pyknotic nuclei, nuclear enlargement
and other nuclear atypia, crystalline inclusions, hyalinization, vacuolar
degeneration, and hydropic degeneration in the cytoplasm. Two more
specific lesion types within the degenerative category included a nuclear
change characterized by a clear halo around the nucleus just inside the
nuclear membrane, and hydropic degeneration with membrane lysis, which was
present only in the digestive epithelium of the hepatopancreas. The
latter lesion was characterized by cytoplasmic cloudy swelling,
dissolution of the plasma membrane, decreased clarity of cellular
contents, and epithelial exfoliation from the underlying basement
membrane.
Proliferative conditions were characterized by increased numbers of
mitotic figures per microscopic field in various tissues, particularly in
the hemopoietic tissue. This increased mitotic activity is probably a
compensatory response to cell injury in these tissues or a host response
to an unspecified endogenous or exogenous mitogen. In no case was there
any evidence of neoplastic transformation of the tissues involved.
Parasitic/infectious conditions included nematode and trematode
helminth infections, microsporidian and other unspecified protozoan
infections, fungal infections, suspected bacterial infections, and
nuclear/cytoplasmic inclusion bodies suggestive of viral infections.
123
-------
3.4.2.1 Cancer magister
Cancer magister were caught in three locations: Bellingham Bay,
Samish Bay and Dungeness Spit (Table 2). No microscopic lesions were
detected in the cardiac and pyloric stomachs, or the gonads of
£. magister. Two main categories of inflammatory conditions appeared
consistently in most of the organs of C. magister: melanized lesions and
unmelanized granulomas. The incidences of these two lesion types varied
considerably among the different collecting sites and from station to
station within each sampling area, revealing no clear distributional
pattern. Incidences of melanized lesions in some tissues (hindgut, eye
and bladder, Table 28; see Table 34 for codes) were found to be somewhat
higher (11% to 40%) in animals from Samish Bay and Dungeness Spit compared
to Bellingham Bay (0% to 7.7%). These same lesions also occurred less
frequently in the hindgut of crabs from Stations 11 and 12 and Stations 23
and 24 in Bellingham Bay. Unmelanized granulomas in the antennal gland
were detected only in animals from Samish Bay and Dungeness Spit.
Unmelanized granulomas in the bladder were not detected in crabs from
Dungeness Spit, but were detected at low incidences at the other two
areas.
A variety of distribution patterns were observed with the other
histopathological conditions. One type of degenerative disorder, hydropic
degeneration/membrane lysis of digestive epithelial cells of the
hepatopancreas, was found only in animals from Bellingham and Samish Bays
(Table 29). This lesion was found to occur at significantly greater
frequencies in crab from Stations 4 and 5 in Bellingham Bay and
significantly lower frequencies in crab from Dungeness Spit. The
incidence of nuclear atypia in digestive epithelial cells of the
hepatopancreas fluctuated broadly among the crab from different areas.
The incidence of proliferative conditions in the hemopoeitic tissue of
C. magister was significantly lower in Samish Bay and higher at
Stations 11 and 12 in Bellingham Bay. The incidences of all inflammatory
conditions (Table 29) were either similar or varied too broadly to
determine clear distributional patterns among the sampling areas. For
124
-------
TABLE 28.
Tissue organ,
lesion type
(no. examined)
Ampulla (N)
Mel lesion
Unmel gran
ANG (N)
Mel
Unmel gran
Degen dis
Memo infil
Bladder (N)
Mel lesion
Unmel gran
Viral inf
Epidermis (N)
Mel lesion
Viral inf
Esophagus (N)
Mel lesion
Hemo infil
Eye (N)
Mel lesion
Unmel gran
Hemo infil
Gill (N)
Mel lesion
Unmel gran
Hemo tissue (N)
Prol cond
HEP (N)
Mel lesion
Unmel gran
Hyd deg
Nuc atypia
Viral inf
Hindgut (N)
Mel lesion
Unmel gran
Hemo infil
Midgut(N)
Mel lesion
Unmel gran
Incidences (%) of Individual Histopathological
Conditions in Cancer magister
Station
11-12
(28)
43
0
(28)
3.6
0
7.1
11
(30)
6.7
10
3.3
(30)
3.3
0
(22)
4.5
0
(28)
3.6
3.6
18
(28)
29
11
(24)
54(H)2
(30)
6.7
6.7
33
20
0
(30)
0(L)
0
13
(29)
10
0(L)
Bellingham
Station
4-5
(27)
30
0
(26)
0
0
7.7
15
(29)
3.4
3.4
3.4
(28)
3.6
0
(25)
8.0
12
(27)
7.4
0
22
(23)
26
0
(23)
35
(29)
3.4
6.9
45(H)
24
3.4
(29)
6.9
3.4
0(L)
(27)
11
11
Bay
Station
23-24
(15)
60
0
(13)
0
0
0
7.7
(17)
0
0
0
(17)
0
0
(12)
0
8.3
(13)
7.7
7.7
15
(17)
29
0
(14)
29
(17)
5.9
0
12
18
0
(16)
0(L)
13
13
(17)
12
0(L)
Samish
Station
1
(26)
46
0
(22)
0
4.5
4.5
14
(28)
11
3.6
0
(29)
6.9
3.4
(17)
5.9
0
(25)
12
4.0
24
(27)
56
7.4
(23)
8.7(L)
(28)
7.1
11
11
50
0
(25)
32
12
12
(25)
20
25
Bay
Station
20
(29)
55
3.4
(24)
8.3
4.2
4.2
17
(28)
21
0
0
(28)
11
3.6
(18)
17
0
(25)
24
0
0
(28)
32
7.1
(26)
12(L)
(28)
7.1
7.1
14
32
0
(29)
31
21
0(L)
(26)
7.7
15
Dungeness
Spit
(11)
64
0
(10)
0
10
0
0
(11)
18
0
10
(10)
0
0
(9)
11
0
(10)
20
20
20
(10)
40
20
(9)
33
(10)
10
20
0(L)
20
0
(10)
40
0
0
(11)
18
36(H)
Definitions of codes for tissue and lesion types in Tables 28-33 are given in
2 Table 34.
L indicates stations classified in the low incidence group, and H indicates
stations in the high incidence group. Significance level for G-statistic was
p <0.05.
125
-------
TABLE 29. Incidences (%) of Major Histopathological Categories
Tissue/organ
lesion type
Ampul 1 a „
Inflam concr
ANC _
Inflam cond^
Bl adder
Inflam cond
Epidermis ,
Inflam conrf^
Esophagus „
Inflam cond
Eye
Inflam cond^
Gill
Inflam cond
HEP -
Inflam cond
Degen dis
Hindgut ,
Inflam cond
Midgut 2
Inflam cond
in Cancer
magister
from Bell
and lJungeness Spit
Bellingham Bay
Station
11-12
43
18
17
3.3
4.5
25
39
13
37
13
10
Station
4-5
30
15
6.7
3.6
12
33
35
6.9
59
10(L)
19
Station
23-24
60
7.7
0(L)3
0
8.3
23
29
5.9(L)
18(L)
19
12
Samish Bay
Station
1
46
H
14
6.9
5.9
36
56
18
48(H)
36
Station
20
55
21
21
11
17
24
36
14
36
41
23
Dungeness
Spit
64
10
18
Definitions of codes for tissue and lesion types in Tables 28-33 are given
2 in Table 34.
Inflammatory conditions consisting of melanized lesions, and unmelanized
3 granulomas and hemocytic infiltration.
L indicates stations classified in the low incidence group, and H indicates
stations in the high incidence group. Significance level for G-statistic
. was p <0.05.
Degenerative disorders consisting of hydropic degeneration/membrane lysis
and nuclear atypia.
11
60
60
30
20
40
55(H)
126
-------
example, the incidences of these conditions in the bladder and
hepatopancreas were significantly lower in crabs from Stations 23 and 24
in Bellingham Bay, significantly lower in the hindgut of crab from
Stations 4 and 5 in Bellingham Bay, but significantly higher in both the
hindgut of crab from Station 1 in Samish Bay and in the midgut of crab
from Dungeness Spit.
3.4.2.2 Cancer gracilis
Cancer gracilis were caught in three areas: Case Inlet, Sinclair
Inlet and Bellingham Bay (Table 2). Histopathological lesion categories
were the same as for C_. magister, except that hydropic degenerative/
membrane lysis in the hepatopancreas was not observed in C. gracilis.
Also no lesions were observed in the cardiac stomach, esophagus, gonad or
midgut ampulla of £. gracilis.
The incidence of melanized lesions in all tissues but eye, bladder,
and hindgut were consistently higher in crabs from Case Inlet than in
those from Sinclair Inlet (Table 30). These lesions were significantly
more prevalent in the antennal gland and gill of crab from Case Inlet.
The incidences of unmelanized granulomas in the antennal gland, bladder,
and midgut were significantly higher in crab from Case Inlet. The
incidences of these lesions in epidermis and pyloric stomach were similar
between the two areas; however, they were more frequently found in
hemopoietic tissue and hindgut of crab from Sinclair Inlet. A distinct
distributional pattern among the sampling sites was not observed for
proliferative conditions in the hemopoietic tissue.
The incidence of combined inflammatory conditions of the gill
(Table 31) was significantly higher in crabs from Case Inlet, and these
lesions were significantly less prevalent in the antennal gland and
epidermis in crab from Sinclair Inlet (Stations 17-20). Incidences of
these conditions in other tissues were generally similar amongst the
sampling areas. The small sample (4 individuals) of C_. gracilis captured
in Bellingham Bay precluded any meaningful comparisons in lesion
incidences with the other areas.
127
-------
TABLE 30.
Tissue organ
lesion type^
(no. examined)
ANG (N)
Mel
Unmel gran
Hemo infil
Oegen dis
Bladder (N)
Mel lesion
Unmel gran
Viral inf
Oegen dis
Epidermis (N)
Mel lesion
Unmel gran
Gill (N)
Mel lesion
Unmel gran
Hemo tissue (N)
Mel lesion
Unmel gran
Prol cond
Eye (N)
Mel lesion
Hemo infil
HEP (N)
Mel lesion
Unmel gran
Nuc atypia
Viral inf
Hindgut (N)
Mel lesion
Unmel gran
Midgut(N)
Mel lesion
Unmel gran
PYL STO (N)
Mel lesion
Unmel gran
Parasite Prev
Viruses
Nematodes
Trematodes
Microsporidans
Fungus
Unidentified
Incidences (%) of Individual Histopathological
Conditions in Cancer gracilis
Case Inlet
Station 11
(16)
29(H)2
35(H)
5.9
18
(23)
26
26(H)
22
26
(23)
52
8.7
(22)
41 (H)
8.7
(20)
15
0
10
(22)
0
4.5
(22)
27
23
32
4.5
(17)
5.9
5.9
(18)
17
50(H)
(19)
11
5.3
40
30
5
0
5
5
0
Bellingham Bay
Station 23-24
(4)
0
0
0
0
(4)
0
25
50
50
(4)
0
0
(4)
0
0
(2)
0
0
0
CO
0
0
(4)
0
25
0
25
(3)
33
0
CO
0
75
CO
0
0
50
50
0
0
0
0
25
Sinclair
Station 17-20
(13)
7.7
0(L)
7.7
7.7
(16)
25
6.3
19
19
(16)
6.3
6.3
(1*)
7.1
14
(16)
0
0
13
(12)
0
8.3
(17)
18
5.6
29
12
(17)
5.9
12
(15)
6.7
20
(12)
0
0
29
29
0
0
0
0
0
Inlet
Station 7
(25)
4.0
8.0
8.0
4.0
(27)
11
3.7
19
26
(28)
41
3.6
(25)
4.0(L)
12
(22)
0
5
4.5
(25)
12
0
(28)
11
18
36
0
(26)
3.8
7.7
(25)
12
20(L)
(28)
3.6
3.6
27
19
0
3.8
3.8
0
3.8
Definitions of codes for tissue and lesion types in Tables 28-33 are given in
Table 34.
L indicates stations classified in the low incidence group, and H indicates
stations in the high Incidence group. Significance level for G-statistic
was p <0.05.
128
-------
TABLE 31. Incidences (%) of Major Histopathological Categories
in Cancer gracilis from Case and Sinclair Inlets
Tissue/organ1
lesion type
ANG
Inflam cond2
Bladder
Inflam cond2
Epidermis
Inflam cond2
Gill
Inflam cond2
Memo tissue
Inflam cond2
Eye
Inflam cond2
HEP
Inflam cond2
Hindgut
Inflam cond2
Midgut
Inflam cond2
PYL STO
Inflam cond2
Case Inlet
Station 11
59
48
52
15(H)
15
4.5
32
12
50
11
Sinclair Inlet
Station 17-ZO
7.7UP
25
6.3(L)
0
0
8.3
18
18
27
0
Station 7
20
15
43
5
5
12
32
12
28
7.1
1 Definitions of codes for tissue and lesion types in Tables 28-33 are
given in Table 34.
2 Inflammatory conditions consisting of melanized lesions, and unmelanized
granulomas and hemocytic infiltration.
3 L indicates stations classified in the low incidence group, and H
indicates stations in the high incidence group. Significance level for
G-statistic was p <0.05.
129
-------
3.4.2.3 Pandalopsis dlspar
Panda lopsis dlspar were caught in two areas: Fourmile Rock - Elliott
Bay dump site vicinity and Dabob Bay (Table 2). No pathologic conditions
were observed in the epidermis, pyloric stomach, or hemopoietic, heart,
and nervous tissues. Some pathological conditions were detected only in
shrimp from Dabob Bay, including melanized lesions of the bladder,
hyalinocytic encapsulation of the eye, and gill, and gonads. Parasitic
infections were also primarily found in shrimp from Dabob Bay. Melanized
lesions and hyalinocytic encapsulation in the gill and hepatopancreas of
shrimp from Dabob Bay were particularly severe. Congestion of the gills
was also detected only in shrimp from Dabob Bay. Proliferative
conditions, hemocytic infiltration and hyalinocytic encapsulation
generally accompanied the melanized lesions in the hepatopancreas. These
conditions were probably associated with unidentified parasitic
infections. Degenerative disorders of the antennal gland were found only
in shrimp from the Fourmile Rock - Elliott Bay dump site vicinity.
Inflammatory conditions in all tissues (except eye) (Table 32) were more
prevalent in shrimp from Dabob Bay.
3.4.2.4 Panda!us platyceros
Pandalus platyceros were also only captured in the Fourmile Rock -
Elliott Bay dump site vicinity and Dabob Bay (Table 2). As with
£. dispar, no pathological conditions were observed in the epidermis,
hemopoietic tissue, nervous tissue, pyloric stomach, eye, or gonad of
P_. platyceros (Table 33). Degenerative disorders of the antennal gland
and heart were detected only in shrimp from the dump site vicinity. On
the other hand, this condition was detected only in the cardiac stomach of
shrimp from Dabob Bay. Increased hyalinocytic phagocytosis was detected
only in the gills of the dump site vicinity specimens, whereas
hyalinocytic encapsulation was more prevalent in the gills and heart of
Dabob Bay specimens. Melanized lesions in the hepatopancreas were also
observed only in shrimp from the dump site vicinity, but the incidence of
these lesions was higher in the gills and heart of shrimp from Dabob Bay.
130
-------
TABLE 32. Incidences (%) of Individual Histopathological
Conditions in Shrimp from Elliott and Dabob Bays
Tissue organ
lesion type*
(no. examined)
ANG (N)
Mel
Hyal encap
Para inf
Oegen dis
Bladder (N)
Prol cond
Hyal encap
Mel lesion
Degen dis
Para inf
Hyal phago
CAS (N)
Pro! cond
Degen dis
Mel lesion
Esophagus (N)
Prol cond
Eye (N)
Hyal encap
Mel lesion
Memo infil
Gill (N)
Mel lesion
Hyal encap
Congestion
Hyal phago
Microan
Para inf
Prol cond
Gonad (N)
Mel lesion
Hyal encap
Heart (N)
Degen dis
Hyal encap
Mel lesion
HEP (N)
Prol cond
Hyal encap
Mel lesion
Hyal phago
Memo infil
Para inf
Viral inf
Panda!opsis dispar
Fourmile Rock-
Elliott Bay2 Oabob Bay
Station 17 Station 15
(16)
6.3
13
0
6.7
(18)
0
5.6
0
17
0
0
(23)
4.3
0
0
(22)
4.5
(22)
0
0
9.1
(22)
0
0
0
0
4.5
0
4.5
(22)
9.1
0
(17)
0
0
0
(23)
8.7
13
13
4.2
8.3
8.3
0
(16)
13
13
13
0
(29)
6.7
14
14
10
6.7
3.4
(30)
3.3
0
6.7
(28)
14
(29)
3.4
3.4
3.4
(30)
23(H)3
13
6.7
6.7
13
13
3.3
(29)
14
6.9
(18)
0
0
0
(30)
20
30
27
17
37
27
3.3
Randal us
platyceros
Fourmile Rock-
Elliott Bay2 Dabob Bay
Station 17 Station 5
(13)
0
0
0
15
(21)
4.8
0
0
4.8
0
0
(21)
5.0
0
0
(20)
10
(20)
0
0
0
(21)
4.7
0(L)
14
4.8
23
0
0
(21)
0
0
(20)
9.1
0
0
(22)
15
9.1
9.1
0
4.5
0
0
(24)
0
0
0
0
(31)
3.2
0
0
6.5
0
0
(31)
19
9.7
0
(30)
10
(31)
0
0
0
(30)
23
37
33
0
30
0
3.3
(30)
0
0
(25)
0
0
4,2
(30)
27
13
0
10
10
9.1
0
Definitions of codes for tissue and lesion types in Tables 28-33 are given in
Table 34.
Dump site vicinity.
L indicates stations classified in the low incidence group, and H indicates
stations in the high incidence group. Significance level for G-statistic
was p O.05.
131
-------
TABLE 33. Incidences (%) of Major Histopathological Categories
for Shrimp from Elliott and Dabob Bays
Tissue/organ^
lesion type
ANG
Inflam cond3
Bladder
Inflam cond3
CAS
Inflam cond0
Eye
Inflam cond0
Gill
Inflam cond3
Vas dis5
Gonad
Inflam cond3
Heart
Inflam cond0
HEP
Inflam cond0
Pandalopsis dispar
Fournrile Rock-
El Hott Bay2
Station 17
13
5.6
9.1
O(L)1*
4.5
9.1
17
Oabob Bay
Station 15
19
6.7
3.4
23
20
37
Panda1 us piatyceros
Fourmile Rock-
El Hott Bay2
Station 17
Oabob Bay
Station 5
5.0(L)
43
9.1
31
63
7.4
18
1 Definitions of codes for tissue and lesion types in Tables 28-33 are given in
Table 34.
2 Dump site vicinity.
3 Inflammatory conditions including me1 am"zed lesions, unmelanized granulomas,
hyolinocytic encapsulation, hyalinocytic phagocytosis and hemocytic infiltration.
L indicates stations classified in the low incidence group, and H indicates
stations in the high incidence group. Significance level for G-statistic was
p<0.05. These designations also apply to Tables 3 and 4.
b Vascular disorders including congestion and microaneurysms.
132
-------
TABLE 34. Lesion and Tissue Codes used in Tables 28-33
AN6 = Antennal gland
CAS = Cardiac stomach
Degen dis = Degenerative disorders
Memo infil = Hemocytic infiltration
H = Incidence is significantly higher at the 0.05 confidence level
Memo tissue = Hemopoietic tissue
HEP = Hepatopancreas
Hyal encap = Hyalinocytic encapsulation
Hyal phago = Hyalinocytic phagocytosis
Inflam cond = Inflammatory conditions
L = Incidence is significantly lower at the 0.05 confidence level
Prol cond = Proliferative conditions
Mel lesion = Mel anized lesions
Microan = Microaneurysms
N = Number of tissues examined
Nuc atypia = Nuclear atypia
Para inf = Parasitic infection
Parasite prev = Parasite incidence
Hyd deg = Hydropic degeneration/membrane lysis
PYL STO = Pyloric stomach
Unmel gran = Unmelanized granuloma
Viral inf = Viral infection
133
-------
3.5 AMPHIPOD BIOASSAY (PERFORMED BY EPA)
3.5.1 Screening Surveys
Results of amphipod bioassays on sediment collected in screening
surveys (August 2 to September 18, 1983) are presented in Figures 32
through 39. Tabular data are presented in Appendix C.
Analyses shown in Table 35 indicate that results of the bioassays for
urban bays were not clearly distinct from results for the baseline bays.
Case Inlet, a baseline bay, showed the lowest mean survival (14.0),
followed by another baseline bay, Dabob Bay, and an urban bay, Port
Gardner - Everett Harbor (both with mean survival of 15.1). Sinclair
Inlet, another urban bay, also demonstrated relatively low mean survival
(15.2). Samish Bay, a baseline bay, and Bellingham Bay, a third urban
bay, showed intermediate mean survival (16.0 and 16.4, respectively).
Sequim Bay, the remaining baseline bay, and Fourmile Rock - Elliott Bay
dump site vicinity, the remaining urban area, demonstrated highest mean
survival (16.9 and 17.2, respectively).
While the four urban bays showed both higher levels of metals and IR
than baseline bays (Table 35), correlation analyses performed on the 207
bioassays from 180 stations only detected a weak negative correlation
(p <0.025) between silver and amphipod survival, and IR and amphipod
survival (p <.10) (Tables 36 and 37). However, a significant negative
correlation (p <0.001) also occurred between percent fines and amphipod
survival, and similarly between percent water and amphipod survival. The
tendency of lower survival became more evident when the exposure sediment
was fine-grained, contained a higher percent water, and contained more
silver. Examining the frequency distribution of amphipod survival in
Table 38 further confirmed that fewer survived in sediment with higher
percent fines and percent water.
Despite the finding that lower survival occurred almost exclusively
in fine-grained sediments, fine-grained sediments did not automatically
mean low amphipod survival. There were 87 cases where 17 or more of 20
amphipods survived. Of these 6 or 6.9% exhibited percent fines >90% and
percent water >;65%.
134
-------
BELLINGHAM BAY
Sampling Locations
FIGURE 32. Results of Amphipod Bloassays on Sediments Collected in
Bellingham bay during Screening Surveys (August 2 to
September 18, 1983).
135
-------
EVERETT HARBOR
Sampling Locations
1/2 0
Nautical Miles
1000 0 1000
FIGURE 33. Results of Amphipod Bioassays on Sediment Collected
in Port Gardner - Everett Harbor During Screening
Surveys (August 2 to September 18, 1983)
136
-------
FIGURE 34. Results of Amphipod Bioassays on Sediments Collected
in the Fourmile Rock - Elliott Bay Dump Site Vicinity
During Screening Surveys (August 2 to September 18,
1983)
137
-------
co
00
SINCLAIR INLET
Sampling Locations
i—i—iii—« «
FIGURE 35. Results of Amphipod Bioassays on Sediments Collected in Sinclair
Inlet during Screening Surveys (August 2 to September 18, 1983)
-------
Nautical Miles
SAMISH BAY
Sampling Locations
FIGURE 36. Results of Amphipod Bioassays on Sediments Collected
in Samish Bay During Screening Surveys (August 2 to
September 18, 1983)
139
-------
CASE INLET
Sampling Locations
FIGURE 37. Results of Amphipod Bioassays on Sediments Collected
in Case Inlet During Screening Surveys (August 2 to
September 18, 1983)
140
-------
DABOB BAY
Sampling Locations
FIGURE 38. Results of Amphipod Bioassays on Sediments Collected from Dabob Bay
During Screening Surveys (August 2 to September 18, 1983)
-------
FIGURE 39. Results of Amphipod Bioassays on Sediments Collected
in Sequim Bay During Screening Surveys (August 2 to
September 18, 1983)
142
-------
TABLE 35. Mean Values (± Standard Deviation) for 1983 Amphipod Survival
CO
and Sediment Characteristics for All Bays (180 Stations)
No. of
Bioassays
with 16 or
or Less
Survivors/
No. of Total No.j
Survivors Bioassays
Overall
Case Inlet
Dabob Bay
Samish Bay
Sequim Bay
Bellingham Bay
Fourmile Rock-,,
Elliott Bay
Port Gardner-
Everett Harbor
Sinclair Inlet
West Beach
(Control)
15.7
14.0
15.1
16.0
16.9
16.4
17.2
15.1
15.2
17.3
± 3.4
± 4.3
± 4.0
± 3.4
± 2.7
± 2.7
± 2.2
± 4.3
± 2.7
± 6.4
92/180
12/20
11/20
11/20
7/20
13/26
10/24
12/25
16/25
4/41
Percent Percent
Water in Fines in Ag Hg Pb IR
Sediment Sediment (ppm) (ppm) (ppm) (ppm)
50.1 ± 14.9 65.2 ± 32.5 0.371 ± 0.477 0.429 ± 0.751 40.6 ± 57.0 477.8 ± 1381.3
64.6 ± 9.7 89.0 ± 19.8 0.299 ± 0.098 0.160 ± 0.043 24.9 ± 6.9 154.4 ± 81.6
41.8 ± 20.1 46.7 ± 40.6 0.079 ± 0.075 0.054 ± 0.042 9.5 ± 7.6 29.3 ± 14.5
46.2 ± 11.0 63.4 ± 37.1 0.078 ± 0.038 0.097 ± 0.052 13.2 ± 6.4 87.5 ± 83.5
53.0 ± 11.0 66.8 ± 26.5 0.136 ± 0.052 0.068 ± 0.029 11. 1± 3.6 96.7 ± 50.6
50.7 ± 11.6 79.6 ± 23.4 0.192 ± 0.108 0.675 ± 0.663 24.9 ± 28.7 566.4 ± 440.1
48.4 ± 14.6 61.3 ± 31.0 0.529 ± 0.194 0.771 ±1.078 88.3 ± 87.4 288.2 ± 218.1
49.7 ± 15.2 52.7 ± 26.6 0.261 ± 0.122 0.158 ± 0.127 29.0 ± 20.6 1627.3 ± 3419.3
47.1 ± 15.3 62.6 1 35.4 1.230 ± 0.763 1.183 ± 1.176 105.6 ± 77.9 652.4 ± 568.2
1 There were some stations with replicate bioassays (actually test replicates). These numbers reflect only stations, not total
number of replicated bioassays. Where test replicates existed, averages were used for that station. Thus, while there were
-207 bioassays, these only represent 180 stations.
Dump site vicinity.
-------
TABLE 36. Correlation Matrix for Numbers of Amphipods
Percent H20
Percent Fines
Ag
Hg
Pb
IR
Surviving
from All
No.
Survivors
0 -0.408
nes -0.331
-0.177
-0.059
-0.105
-0.123
(1983) and Sediment
Bays (180 Stations)
Percent
H20
0.827
0.273
0.095
0.140
0.223
Percent
Fines
0.283
0.128
0.104
0.002
Characteristics
Ag
0.574
0.677
0.146
Hg
0.871
0.161
Pb
0.219
TABLE 37.
No. Survivors
Percent H20
Percent Fines
Ag
Hg
Pb
IR
Results of Regression Analyses for Numbers
of Amphipods Surviving (1983) and Sediment
Characteristics from all Bays (180 Stations)
p-level
-0.408
-0.331
-0.177
-0.059
-0.105
-0.123
0.
0.
0.
0.
0.
0.
166
110
031
003
on
015
P
P
P
P
<.001
<.001
<.025
-
-
<.100
144
-------
TABLE 38. Mean Percent H20 and Percent Fines
at Various Survivor Levels
No. Survivors
3
4
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
No. of Cases
1
1
1
1
7
6
2
8
8
13
9
25
22
21
31
31
20
Percent
H20
70.05
66.73
67.10
66.73
62.72
67.19
45.46
58.29
54.56
56.88
55.67
48.77
51.28
48.39
45.12
45.16
41.77
Percent
Fines
72.93
64.28
70.30
64.28
85.56
90.21
81.87
75.66
72.33
79.67
66.14
68.72
68.30
70.48
52.36
57.33
48.47
145
-------
3.5.2 Detailed Surveys
Results of amphipod bioassays collected during detailed surveys
(April 23 to May 29, 1984) are presented in Figures 40 through 47.
Tabular data are presented in Appendix D.
Results of analyses, shown in Tables 39 through 44, demonstrated that
statistically significant (p <0.05) differences among mean amphipod
survival and reburial occurred in all assays (1 through 6). Subsequent
analyses applying Dunnett's procedure demonstrated statistically
significant (p <0.05) differences among the station means as summarized in
Table 45. These latter analyses were performed with survival data and not
reburial data. By inspection, however, reburial results parallel survival
results with only minor exception.
The analyses shown in Table 46 indicated the lack of a significant
batch effect: P(x2 >. 2.21) = 0.81. Each batch of amphipods responded
similarly to the reference toxicant PCP, thus eliminating handling or test
organism condition as a cause of the differences among treatments
observed.
Based on these data, and the data presented in Table 47, the urban
bays were not clearly distinct from baseline bays. Of the urban bays,
Port Gardner - Everett Harbor and Sinclair Inlet exhibited the lowest mean
survival (12.3 and 13.5, respectively). Intermediate survival was
observed in Bellingham Bay, while the highest mean survival of amphipods
in urban bay sediments occurred in the Fourmile Rock - Elliott Bay dump
site vicinity. Of the baseline bays, Case Inlet and Samish Bay showed the
lowest mean survival (13.0 and 13.3, respectively), followed by Dabob Bay
(16.1) and Sequim Bay (17.0).
Examination of the correlation matrix in Table E.2 revealed that
amphipod survival correlated with several physical and chemical
parameters. The trend toward lower survival became more evident when the
sediments were finer grained and contained more water and more organic
contaminants as exemplified by PCB-1254, aromatic hydrocarbons, and IR.
146
-------
Squall cum Cr*.
Waterway .
BelUngham Bay
Station
3.
4.
5.
7.
11.
12.
23.
24.
kMortality
Survival
% Survival
96
83
66
95
83
89
42
89
Whatcom Cr>.
Wate rwav '
Starr RQ
24 J
23.
iellingham
48-44
FIGURE 40. Results of Amphipod Bioassays on Sediments Collected
in Bellingham Bay During Detailed Surveys (April 23
to May 29, 1984)
147
-------
EVERETT HARBOR
Sampling Locations
1/2 0
FIGURE 41. Results of Amphipod Bioassays on Sediments Collected
in Port Gardner - Everett Harbor During Detailed
Surveys (April 23 to May 29, 1984)
148
-------
Samp
MAGNOLIA BL
Four-mile Rock
Lion BAY
ng Locations
1/2
tical Miles
F
47 - 38 N
24.
ELLIOK BAY
Station
Q
Survival
07
47'- 36'N
10.
12.
17.
20.
22.
23.
24.
86
34
92
83
86
38
37
122*-
26 W
122 -
FIGURE 42. Results of Amphipod Bioassays on Sediments Collected
in the Fourmile Rock - Elliott Bay Dump Site Vicinity
During Detailed Surveys (April 23 to May 29, 1984)
149
-------
en
o
Mortality
Survival
SINCLAIR INLET
Sampling Locations
Station X Survival
6. 55
7. 81
8. 62
14. 57
17. 74
18. 68
19. 81
20. 62
7. 8.
Sinclair Inlet
Nautical Miles
0 1000
FIGURE 43. Results of Amphipod Bioassays on Sediments Collected in Sinclair
Inlet During Detailed Surveys (April 23 to May 29, 1984)
-------
Nautical Miles
SAMISH BAY
Sampling Locations
Station I Survival
1. 69
3. 53
7. 75
20. 69
FIGURE 44. Results of Amphipod Bioassays on Sediments Collected
in Samish Bay During Detailed Surveys (April 23 to
May 29, 1984)
151
-------
en
ro
DABOB BAY
Sampling Locations
FIGURE 45. Results of Amphipod Bioassays on Sediments Collected from Dabob
Bay During Detailed Surveys (April 23 to May 29, 1984)
-------
CASE INLET
Sampling Locations
Station X Survival
1. 72
11. 72
15. 57
17. 59
FIGURE 46. Results of Amphipod Bioassays on Sediments Collected
from Case Inlet During Detailed Surveys (April 23 to
May 29, 1984)
153
-------
>"\ J
FIGURE 47. Results of Amphipod Bioassays of Sediments Collected
from Sequim Bay During Detailed Surveys (April 23 to
May 29, 1984)
154
-------
TABLE 39. Summary of Statistical Analyses (HOMOV, ANOVA, SNK, Dunnett's
Test) of Survival and Reburial Data for Rhepgxynius abronius
Exposed to Puget Sound Sediments (Assay No. 1 - Bellingham
Bay [B-l], Samish Bay [B-2])
SURVIVAL
' max * max.05 (9,4)
X2c = 16.15 X2.05(8) = 15.507
C = 0.267 C-nfln ._, = 0.3584
ANOVA; EMS = 2.78889 F
.001(8,36)
6 F.05(8,36)
SNK; B-1 = Bellingham Bay B-2 = Samish Bay
Sample B-2 S-3 B-l S-5 B-2 S-1 B-2 S-20 B-2 S-7 B-l S-4 B-l S-7 B-l S-3 Cont.
- 10.6 13.2 13.8 13.8 15.0 16.6 19.0 19.2 19.8
6 3 5 8 7 2 419
PUNNETTS; * * * * * * N.S N.S Cont.
10.6 13.2 13.8 13.8 15.0 16.6 19.0 19.2 19.8
* Significant p<0.05
REBURIAL
HOMOV: F = 31.5
max
= 18.6
C = 0.269
ANOVA: EMS = 2.600
F
.05<8,36) '
SNK:
Sample B-2 S-3 B-l S-5 B-2 S-1 B-2 S-20 B-2 S-7 B-l S-4 B-l S-7 B-l S-3 Cont.
- 10.4 13.0 13.6 13.8 15.0 16.2 19.0 19.2 19.8
6 3 _ 5 _ 8 _ 7 2419
* * * * * N.S N.S Cont.
13.0 13.6 13.8 15.0 16.2 19.0 19.2 19.8
* Significant p<0.05
155
-------
TABLE 40. Summary of Statistical Analyses (HOMOV, ANOVA, SNK, Dunnett's
Test) of Survival and Reburial Data for Rhepgxynius abronius
Exposed to Puget Sound Sediments (Assay No. 2 - Bellingham
Bay [B-l], Port Gardner - Everett Harbor [B-5])
SURVIVAL
=12'875 F™,ax.05(9,4> - *1'1
X2C = 9.758 X2>05(8) = 15.507
EMS = 4.25556
F =22'735 F.001(8,36, ' 3'7*
§NK: B-l = Bellinghara Bay B-4 = Port Gardner - Everett Harbor
Sample B-l S-23 B-4 S-4 B-4 S-l B-4 S-3 B-4 S-2 B-l S-ll B-l S-24 B-l S-12 Cont.
- 8.4 8.4 8.8 13.4 14.2 16.6 17.8 17.8 19.4
3 8 5 7 6 1 4 2 9_
DUNNETTS; * * * * * N.S N.S N.S Cont.
8.* 8.4 8.8 13.4 14.2 16.6 17.8 17.8 19.4
* Significant p<0.05
REBURIAL
Wtm: F = 14.125
max
X2C = 8.95
ANQVA; EMS = 4.2444
F = 29.2147
SNK;
Sample B-l S-23 B-4 S-4 B-4 S-l B-4 S-3 B-4 S-2 B-l S-ll B-] S-24 B-l S-12 Cont.
~ 5.6 7.8 8.4 12.6 14.0 16.4 17.4 17.8 19.4
_8 5 3 7 6 1 4 2 9_
DWNETT5; * * * * * N.s N>s N>s Cont>
5.6 7.8 8.4 12.6 14.0 16.4 17.4 17.8 J9.4
* Significant p<0.05
156
-------
TABLE 41. Summary of Statistical Analyses (HOMOV, ANOVA, SNK, Dunnett's
Test) of Survival and Reburial Data for Rhepoxynius abronius
Exposed to Puget Sound Sediments (Assay No. 3 - Port Gardner •
Everett Harbor [B-4], Fourmile Rock - Elliott Bay Dump Site
Vicinity [B-5])
SURVIVAL
HOMOV;
F
max
2
X C
=- 32.33
= 12.7665
max. 05(9, 4) = 41.1
2
X „,-,„, = 15.507
ANOVA; EMS =
F = 41.8923
SNK; B-4 Port Gardner - Everett Harbor B-5 Fourmile Rock - Elliott Bay
Sample B-4 S-5 B-4 S-ll B-5 S-12 B-5 S-10 B-5 S-9 B-4 S-6 B-4 S-7 B-5 S-17 Cont.
2.8 14.8 16.8 17.2 17.4 17.8 18.0 18.4 19.4
1 2 3 :4 5 6 7 89
DUNNETTS; * * N.S N.S N.S N.S N.S N.S Cont.
2.8 14.8 16.8 17.2 17.4 17.8 18.0 18.4 19.4
* Significant p<0.05
REBURIAL
HOMOV; F = 27.667
oax
X C = 11.2541
ANOVA; EMS = 2.9556
F = 45.4812
SNK;
Sample B-4 S-5 B-4 S-ll B-5 S-12 B-5 S-10 B-5 S-9 B-4 S-6 B-4 S-7 B-5 S-17 Cont.
- 2.4 14.6 16.8 17.2 17.4 17.8 18.0 18.4 19.4
14 7652389
DUNNETTS; * * N.S N.S N.S N.S N.S N.S Cont.
2.4 14.6 16.8 17.2 17.4 17.8 18.0 18.4 19.4
* Significant p<0.05
157
-------
TABLE 42. Summary of Statistical Analyses (HOMOV, ANOVA, SNK, Dunnett's
Test) of Survival and Reburial Data for Rhepoxynius abronius
Exposed to Puget Sound Sediments (Assay No. 4 - Fourmile Rock
Elliott Bay Dump Site Vicinity [B-5], Sinclair Inlet [B-6])
SURVIVAL
HOMOV;
F = 62.5
max
X^C = 18.4098
ARCS IN /"xc
Fmax.05(9,4)
X .05(8)
C. 05(9,4)
41.1
15.507
0.3584
F - 42.0901
max
= 10.6197
C = 0.267
ANOVA: (on transformed data)
EMS = 66.23021
F = 8'94U F.001(8,36, =3'm
SNK; B-5 Fourmile Rock - Elliott Bay - B-6 Sinclair Inlet
Sample B-6 S-7 B-6 S-14 B-6 S-8 B-6 S-7 B-5 S-20 B-5 S-22 B-5 S-23 B-5 S-24 Cent,
- 11.0 11.4 12.4 16.2 16.6 17.2 17.6 17.4 19.0
- Arcsin
x
*8.0 49.1 52.2 64.9 66.1 68.1 70.4 71.7 78.6
5 _ 8 _ 7 _6 _ ] _ 2 _ 2 _ 4 9
PUNNETTS; * * * N.S N.S N.S N.S N.S Cont.
48.0 49.1 52.2 64.9 66.1 68.1 70.4 71.7 78.6
* Significant p<0.05
_ REBURIAL _
HOMOV; F = "0" in variance
^~~~ max
X2C = 12.33
C = 0.296
ANOVA; EMS = 4.6222
F = 9.4700
SNK;
Sample B-6 S-7 B-6 S-14 B-6 S-8 B-6 S-7 B-5 S-20 B-5 S-22 B-5 S-23 B-5 S-24 Cont
11.0 11.4 12.4 16.2 16.6 17.0 17.2 17.6 19.0
58761 2439
x
DUNNETTS; * * * N.S. N.S N.S N.S N.S Cont.
11.0 11.4 12.4 16.2 16.6 17.0 17.2 17.6 19.0
* Significant p<0.05
158
-------
TABLE 43. Suranary of Statistical Analyses (HOMOV, ANOVA, SNK, Dunnett's
Test) of Survival and Reburial Data for Rhepoxynius abrom'us
Exposed to Puget Sound Sediments (Assay No. 5 - Sinclair Inlet
[B-6], Dabob Bay [B-7])
SURVIVAL
"" max = "0" in variance
X2C = 12.7223 X2 - 15.507
ANOVA; EMS = 4. 57778
C.05(9,4) = 0.3584
F = 7.8871 F nclo ,,t = 3.794
.05(8,36)
SNK; B-6 = Sinclair Inlet B-7 = Dabob Bay
Sample B-6 S-20 B-7 S-15 B-6 S-18 B-6 S-17 B-7 S-7 B-6 S-19 B-7 S-l B-7 S-5 Cont.
- 12.4 12.8 13.6 14.8 14.8 16.2 18.0 18.6 20.0
482 173569
DUNNETTS; * * * * * * N.S N.S Cont.
12.4 12.8 13.6 14.8 H.8 16.2 18.0 18.6 20.0
* Significant p<0.05
REBURIAL
HOMOV: F, = "0" in variance
~~~~~ max
X C = 12.8929
C = 0,216
ANOVAj EMS = 4.4778
F = 7.866
SNK;
Sample B-6 S-20 B-7 S-15 B-6 S-18 B-6 S-17 B-7 S-7 B-6 S-19 B-7 S-l B-7 S-5 Cont.
- 12.4 12.8 13.6 H.8 14.8 15.4 18.0 18.4 20.0
Jt 8 2 1 7 3_ 5 69
DENNETTS; * * * * * * N.S N.S Cont.
12.4 12.8 13.6 14.8 H.8 15.4 18.0 18.4 20.0
* Significant p<0.05
159
-------
TABLE 44. Summary of Statistical Analyses (HOMOV, ANOVA, SNK, Dunnett's
Test) of Survival and Reburial Data for Rhepoxynlus abronius
Exposed to Puget Sound Sediments (Assay No. 6 - Sequim Bay L8-3],
Case Inlet [B-8])
SURVIVAL
HOMOV; F = 7.800 F n , =41.1
max max.05(9,4)
X*C = 5.'2853 X2.05(8) = 15.507
ANOVA: EMS =4.48889 F.001(8,36) = *>™
SNK: B-8 = Case Inlet B-3 = Sequim Bay
Sample B-8 S-15 B-8 S-17 B-8 S-l B-8 S-ll B-3 S-20 B-3 S-14 B-3 S-18 B-3 S-17 Cont.
11.* 11.8 14.4 14.4 14.6 17.6 17.8 18.0 19.0
7 8 56 4 1 3 29
DUNNETTSt * * * * * N.S N.S N.S Cont.
11.* 11.8 14.4 14.4 14.6 17.6 17.8 18.0 19.0
* Significant p<0.05
REBURIAL
HOMOV! F = 6.7
~ max
X2C = 4.3814
ANOVA: EMS = 4.4444
F = 8.8475
SNK;
Sample B-8 S-15 B-8 S-17 B-8 S-l B-8 S-ll B-3 S-20 B-3 S-14 B-3 S-18 B-3 S-17 Cont.
- 11.2 11.8 14.4 14.4 14.4 17.6 17.8 17.8 19.0
7 8*5612 39
DUNNETTS;
* * * * * N.S N.S 'N.S Cont.
11.2 11.8 14.4 14.4 14.4 17.6 17.8 17.8 19.0
* Significant p<0.05
160
-------
TABLE 45. Significance of Amphipod Survival Data by Bay
and Station for Sediments Collected During
Detailed Surveys April 23 to May 29, 1984
Location Significant! Not Significant* Inconclusive3
Bellingham Bay
B-l S-5
B-l S-23
B-l S-3
B-l S-7
B-l S-12
B-l S-24
B-l S-4
B-l S-ll
Samish Bay B-2 S-l
B-2 S-3
B-2 S-7
B-2 S-20
Port Gardner-Everett Harbor B-4 S-l B-4 S-6
B-4 S-2 B-4 S-7
B-4 S-3
B-4 S-4
B-4 S-5
B-4 S-ll
Fourmile Rock-Elliott Bay** B-5 S-9 B-5 S-10
B-5 S-17 B-5 S-12
B-5 S-23 B-5 S-20
B-5 S-24 B-5 S-22
i <16.2 surviving was significant (p <0.05).
2 >17.2 surviving was not significant (p >0.05).
3 Values between 16.2 and 17.2 are classified as inconclusive.
** Dump site vicinity.
161
-------
TABLE 45. Significance of Amphipod Survival Data by Bay
and Station for Sediments Collected During
Detailed Surveys April 23 to May 29, 1984
(Continued)
Location Significant1 Not Significant2 Inconclusive3
Sinclair Inlet B-6 S-6 B-6 S-7
B-6 S-8 B-6 S-19
B-6 S-14
B-6 S-17
B-6 S-18
B-6 S-20
Dabob Bay B-7 S-7 B-7 S-l
B-7 S-15 B-7 S-5
Sequim Bay B-3 S-20 B-3 S-14
B-3 S-17
B-3 S-18
Case Inlet B-8 S-l
B-8 S-ll
B-8 S-15
B-8 S-17
162
-------
TABLE 46. Chi-Square 2x6 Analysis to Test for
Batch Effect in Amphipod Test Populations
#1
132
48
180
2
127
53
180
3
128
52
180
4
129
51
180
5
125
55
180
6
120
60
180
Total
761
319
1080
Batch
Alive
Dead
Total
X2 = 2-21
P (X2 > 2.21) = 0.81
O
The results of the 1984 bioassay series generally reflected the
results of the 1983 bioassay series. Assay of the Fourmile Rock - Elliott
Bay dump site vicinity and Sequim Bay sediments resulted in the highest
survival in both surveys. Port Gardner - Everett Harbor and Case Inlet
consistently demonstrated the lowest survival, indicating good agreement
between the independent series. Actually, the results of the 1984
bioassay series successfully reproduced the results of the 1983 series at
the screening level 63% of the time.
Statistically speaking (see Table 48), with approximately 50% of the
bioassays in each survey resulting in a "hit" (16 or fewer survivors), a
totally random procedure would be expected to yield equal quantities in
four categories: (1) both 1983 and 1984 a "hit"; (2) both 1983 and 1984
not "hits"; (3) 1983 a "hit," 1984 not; and 4) 1983 not, 1984 a "hit." In
fact, the procedure yielded 30 of 48 (62.5%) correct pairs. Comparing the
procedure with random chance yields a chi-square value of 7.17 with
3 degrees of freedom (df) which is significant at the 0.10 level to reject
the hypothesis of no difference. If stations with survival from 15.1 to
16.9 are rejected as being inconclusive, then 35 cases remain (Table 49).
The procedure then matched on 22 of the 35 (62.9%). Again, testing the
procedure against random chance yielded a chi-square value of 6.03 with
3 df, which rejects the null hypothesis of no difference at the 0.25
163
-------
TABLE 47. Mean Values (± Standard Deviation) for 1984 Amphipod Survival
and Sediment Characteristics for
No. Bioassays
with 16 or Less Percent Percent
No. of Survivors/Total Water in Fines in
Survivors No. Bioassays Sediment
Overall
Case Inlet
Dabob Bay
Samish Bay
Sequim Bay
Bellingham Bay
Fourmile Rock-
Elliott Bay1
Port Gardner-
Everett Harbor
Sinclair Inlet
West Beach
(Control )
14.8 ± 3.3
13.0 ± 1.6
16.1 ± 2.7
13.3 ± 1.9
17.0 ± 1.6
16.1 ± 3.6
17.3 ± 3.5
12.3 ± 5.2
13.5 ± 2.0
19.4 ± 4.2
25/48
4/4
2/4
4/4
1/4
2/8
0/8
6/8
6/8
0/30
57.6
64.6
47.1
54.6
63.3
59.6
49.4
61.6
60.5
± 10.1
± 8.7
± 15.8
± 3.9
± 2.8
± 7.5
± 11.4
± 9.7
± 5.3
Sediment
69.5 ± 21.4
72.0 ± 22.5
45.6 ± 32.0
83.6 ± 3.1
77.6 ± 9.3
86.7 ± 13.9
52.7 ± 23.9
63.4 ± 12.4
74.7 ± 15.8
All Bays (48 Stations)
Ag Hg
(ppm)
0.609 ± 0.800
0.329 ± 0.189
0.093 ± 0.090
0.114 ± 0.010
0.203 ± 0.027
0.249 ± 0.058
0.552 ± 0.213
0.299 ± 0.098
2.186 ± 0.797
(ppm)
0.850
0.127
0.064
0.100
0.770
1.025
1.540
0.239
2.113
± 1.410
± 0.024
± 0.027
± 0.007
± 0.010
± 0.446
± 2.810
± 0.109
± 1.050
Pb
(ppm)
66.7 ±
26.3 ±
9.9 ±
14.9 ±
13.3 ±
23.8 ±
134.1 ±
40.1 ±
170.3 ±
90.8
5.0
4.9
0.7
1.3
8.6
155.2
15.8
68.1
IR
(ppm)
743.1 ± 967.2
91.2 ± 32.3
45.1 ± 24.8
91.8 ± 9.1
88.9 ± 22.2
924.9 ± 492.0
304.0 ± 182.8
1852.0 ±lit90j&
1219.3 ± 923.2
1
Dump site vicinity.
-------
TABLE 47. Mean Values (± Standard Deviation) for 1984 Amphipod Survival
and Sediment Characteristics for All Bays (48 Stations)
(Continued)
en
Overal1
Case Inlet
Dabob Bay
Samish Bay
Sequlm Bay
Bellingham Bay
Fourmile Rock-
Elliott Bay
Port Gardner
Everett Harbor
Sinclair Inlet
West Beach (Control)
No. of
Survivors
H.8 ± 3.3
13.0 ± 1.6
16.1 ± 2.7
13.3 ± 1.9
17.0 ± 1.6
16.1 ± 3.6
17.3 ± 3.5
12.3 ± 5.2
13.5 ± 2.0
19.4 ± 4.2
No. Bioassays
with 16 or Less
Survivors/Total
No. Bioassays
25/48
4/4
2/4
4/4
1/4
2/8
0/8
6/8
6/8
0/30
PCB-1254
ppb
315.7 ± 1441.1
Undetected
Undetected
Undetected
Undetected
48.8 ± 26.9
319.0 ± 411.1
316.7 ± 252.9
578.5 ± 364.3
Total Aromatics
ppb
5387.1 ± 6882.8
Undetected
Undetected
1252.0 ± 1325.4
Undetected
2591.9 ± 2384.5
4141.2 ± 5153.4
8594.6 ± 7538.5
8288.3 ± 10279.0
Dump site vicinity.
Mean of detectable values only.
-------
TABLE 48. Comparison of Procedure with Random
Chance, All 48 Stations
1983-1984 1983-1984 1983-hit 1983-not
both hits not hits 1984-not 1984-hit
EPA 19 11 12 6
Random 12 12 12 12
X2 = 7.17 3 df sig. a = 0.10
TABLE 49. Comparison of Procedure with Random Chance,
13 Stations with Inconclusive Results (Mean
Survival Between 15.1 and 16.9) Removed
1983-1984 1983-1984 1983-hit 1983-not
both hits not hits 1984-not 1984-hit
EPA 15 776
Random 8.75 8.75 8.75 8.75
X2 = 6.03 3 df sig. a = 0.25
166
-------
level. The procedure is especially accurate in duplicating the situation
where both 1983 and 1984 results showed "hits" at a given station.
3.6 OYSTER LARVAL BIQASSAY (PERFORMED BY MRL)
3.6.1 Screening Surveys
Results of oyster larval bioassays in selected sediments collected
during screening surveys are presented in Table 50. A summary of the
experimental conditions for each bioassay is found in Appendix D.
Although rigorous statistical comparisons were not possible (nor
intended), oyster larval bioassay of sediments from all urban bay
stations, with the exception of Station No. 6 at the Fourmile Rock -
Elliott Bay dump site vicinity, indicated clear evidence of a
concentration effect. Increased abnormality occurred generally at the
higher test concentrations (10 and 100 g/L). However, the Fourmile Rock -
Elliott Bay dump site Station No. 6 and the only baseline bay station,
Sequim Bay No. 14, showed virtually no differences among concentrations.
3.6.2 Detailed Surveys
Results of oyster larval bioassays on sediments collected in detailed
surveys (April 23 to May 29, 1984) are presented in Tables 51 through 56.
A summary of the experimental conditions for each bioassay is presented in
Appendix D.
Analyses shown in Table 57 demonstrated that statisically significant
differences (p<0.05) from controls for the following stations: Stations
11, 12, 23, and 24 in Bellingham Bay; Stations 6, 7, 14, 19, and 20 in
Sinclair Inlet; and Stations 1 and 7 in Dabob Bay (Tables 58 and 59).
Examination of the correlation matrix in Appendix E, shows that for
all 48 oyster larval bioassays, no significant (p <0.05) relationships
existed between chemical variables and percentages of abnormal larvae.
Examination of the correlation matrix in Appendix E also shows a negative
correlation (-0.29) between percent abnormal oyster larvae and amphipod
167
-------
TABLE 50. Effects of Sediment Concentration on Oyster Larval Development Measured as Percent
Abnormals (Assays performed August 24 to September 24, 1983 on Fresh Puget Sound
Sediments)
00
Sediment
Concentration
(9/L)
Control
0.01
0.10
1.00
10.00
100.00
Test date.
Dump site vicinity.
Sediment control.
Port Gardner-
Everett Harbor
Station. 2
8-24-831
13.3
6.3
7.3
x = B~79
9.5
2.6
8.8
x = 579
10.7
0.0
12.7
x = ~77ff
9.6
2.6
15.3
12.3
14.6
16.7
TT75
44.0
23.0
8.3
7BTT
Fourmi1e Rock-
El liott Bay^
Station,2
8-27-831
21.9
17.1
11.3
x = TC7E
8.9
20.8
27.4
x = TU70
14.0
24.0
32.5
x = 2T75
17.9
38.6
27.1
x = 1775
34.4
26.7
36.5
x = 7775
35.3
35.9
22.7
x = 3TTT
Fourmi 1 e Rock-
En iott Bay2 Sinclair
Station. 6 Station.1
8-27-831
x
x
X
X
X
X
11.1
6.8
5.4
= 777
15.1
28.6
6.4
= T577
7.9
16.6
3.2
= ~577
8.6
0.0
13.3
= -77J
8.3
3.8
16.9
= ~575
9.4
10.7
7.7
= ~577
8-31 -83*
29.9
16.6
16.8
x = 7T7T
14.5
30.2
13.9
x = T?75
24.2
30.2
9.1
x = 7T7T
31.8
21.4
31.0
x = :ZF70"
39.6
30.9
30.9
5 = 3T7ff
25.0
46.5
58.3
x = 4T7?
Sinclair
Station,?
8-31 -831
12.9
10.9
10.3
x = TT7J
14.5
4.4
6.4
x = ~ff74"
11.3
10.6
13.4
x = TT77
17.2
26.1
11.5
x = TC7Z
36.0
18.8
27.3
x = 777J
34.7
39.5
31.3
x = 3T7f
Sequim3
Station, 14
9-22-831
15.4
10.0
10.0
x = TT76
11.6
11.5
7.0
x = TTJTff
21.0
0.0
7.0
x = ~97J
29.6
15.4
20.4
x = 2T78
15.1
11.9
8.8
x = TT75
12.8
19.1
8.8
x = TJ75
-------
TABLE 51. Effects of Puget Sound Sediments on Oyster Larval Development Measured
as Percent Abnormals (Assay No. 1 Performed April 26 to 28, 1984)
Samlsh Bay
10
Station
1
3
7
20
Control
Rep.
A
B
C
D
Total
'A
B
C
D
Total
A
B
C
D
Total
A
B
C
D
Total
A
B
C
D
No.
Scored
16
49
22
113
200
25
57
19
0
101
17
7
73
35
132
42
120
40
13
215
127
175
198
214
No.
Abnormal
0
1
2
3
0
1
0
0
1
0
1
1
1
3
1
1
2
0
4
8
3
17
6
Percent
Abnormal
3.0
0.9
2.2
1.8
BelUnqham Bay
Station
3
4
5
7
Control
B
C
D
Total
A
B
C
D
Total
A
B
C
D
Total
A
B
C
D
Total
E
F
G
H
Total
No.
Scored
118
64
10
66
258
109
36
58
157
360
34
41
109
34
218
77
33
22
74
206
3
3
173
215
1108
No.
Abnormal
2
0
1
0
3
0
0
3
6
9
2
0
1
3
6
0
0
0
1
1
3
2
2
6
47
Percent
Abnormal
1.2
2.5
2.8
0.5
4.21
Percent abnormal of eight control test replicates.
-------
TABLE 52. Effects of Puget Sound Sediments on Oyster Larval Development Measured
as Percent Abnormals (Assay No. 2 Performed May 10 to 12, 1984)
Bellingham Bay
Port Gardner - Everett Harbor
Station
11
12
23
24
Control
No.
Rep. Scored
~~7T
B
C
D
Total
A
B
C
D
Total
A
B
C
D
Total
A
B
C
D
Total
A
B
C
D
49
19
7
42
117
19
45
8
59
131
1
01
01
01
1
165
0
199
12
376
114
178
156
154
No.
Percent
Abnormal Abnormal
4
4
1
2
11
7
9
0
4
20
1
0
0
0
1
35
0
1
3
39
2
2
0
2
9.4
15.2
100.0
10.4
No.
Station Rep. Scored
1 A
B
C
D
2
3
4
Control
Total
A
B
C
D
Total
A
B
C
D
Total
A
B
C
D
Total
E
F
G
H
Total
23
22
33
25
103
7
3
10
0
20
12
7
26
80
125
6
27
85
6
124
276
235
141
128
1382
No.
Abnormal
1
0
1
1
3
0
0
0
0
0
1
0
1
0
2
0
2
0
0
2
2
1
1
0
10
Percent
Abnormal
2.9
0.0
1.6
1.6
0.72
1 Total mortality.
2 Percent abnormal of eight control test replicates,
-------
TABLE 53. Effects of Puget Sound Sediments on Oyster Larval Development Measured
as Percent Abnormal s (Assa,
Port Gardner - Everett Harbor
Station
5
6
7
11
Control
No.
Rep. Scored
B
C
D
Total
A
B
C
D
Total
A
B
C
D
Total
A
B
C
D
Total
A
B
C
D
46
54
54
55
209
211
31
129
247
618
155
36
124
166
481
35
292
126
87
540
430
984
3160
576
No.
Abnormal
22
20
6
6
54
16
1
4
13
34
16
5
5
5
31
7
8
4
5
24
24
103
60
69
Percent
Abnormal
25.8
5.5
6.4
4.4
Fourmlle Rock -
No.
Station Rep. Scored
9 A
B
C
D
10
12
17
Control
Total
A
B
C
D
Total
A
B
C
D
Total
A
B
C
D
Total
E
F
G
H
Total
362
68
69
213
712
5
0
185
66
256
437
217
313
47
1014
42
517
465
213
1237
625
1154
676
614
8219
• Elliott Bay1
No. Percent
Abnormal Abnormal
5
1
10
38
54
0
0
8
3
11
18
5
150
4
177
3
19
327
4
353
23
37
18
14
348
7.6
4.3
17.5
28.5
4.22
1 Dump site vicinity.
2 Percent abnormal of 8 control test replicates.
-------
TABLE 54. Effects of Puget Sound Sediments on Oyster Larval Development Measured
as Percent Abnormals (Assay No. 4 Performed May 17 to 19, 1984)
Four-mile Rock - Elliott Bay1
Sinclair Inlet
ro
Station
20
22
23
24
Control
No.
Rep. Scored
A
B
C
D
Total
A
B
C
D
Total
A
B
C
D
Total
A
B
C
D
Total
A
B
C
D
110
295
269
262
936
162
184
285
393
1024
68
221
25
243
557
139
118
167
239
663
836
753
764
1422
No.
Abnormal
2
3
5
2
12
0
3
0
5
8
4
6
2
5
17
1
6
1
4
12
4
10
9
35
Percent
Abnormal
1.3
0.8
3.1
1.8
No.
Station Rep. Scored
6 A
B
C
D
7
8
14
Control
Total
A
B
C
D
Total
A
B
C
D
Total
A
B
C
D
Total
E
F
G
H
Total
58
114
82
29
283
217
106
97
203
623
219
216
170
219
824
74
62
218
243
597
430
381
395
325
5306
No.
Abnormal
2
5
8
3
18
14
8
10
10
42
7
4
6
12
29
8
8
13
11
40
4
6
6
4
78
Percent
Abnormal
6.4
6.7
3.5
6.7
1.52
1 Dump site vicinity.
2 Percent abnormal of eight control test replicates,
-------
00
TABLE 55. Effects of Puget Sound Sediments on Oyster Larval Development Measured
Dabob Bay
as Percent Abnormal s (Assay
Sinclair Inlet
Station
17
18
19
20
Control
No.
Rep. Scored1
B
C
D
Total
A
B
C
D
Total
A
B
C
D
Total
A
B
C
D
Total
A
B
C
D
200
200
200
200
800
200
200
200
200
800
200
200
200
200
800
200
200
200
200
800
200
200
200
200
No.
Abnormal
23
24
6
16
69
23
8
8
16
55
51
57
23
47
178
30
28
18
19
95
6
0
3
5
Percent
Abnormal
8.6
6.9
22.3
11.9
Station
1
5
7
15
Control
^
B
C
D
Total
A
B
C
D
Total
A
B
C
D
Total
A
B
C
D
Total
E
F
G
H
Total
No.
Scored1
200
200
200
200
800
61
200
200
200
661
200
200
200
200
800
200
200
200
200
800
200
200
200
200
1600
No.
Abnormal
22
36
12
11
81
11
9
9
2
31
24
40
17
7
88
23
15
6
19
63
3
4
6
4
31
Percent
Abnormal
10.1
4.7
11.0
7.9
1.92
1 Rescored, see page 179.
2 Percent abnormal of eight control test replicates.
-------
TABLE 56. Effects of Puget Sound Sediments on Oyster Larval Development Measured
as Percent Abnormals (Assay No. 6 Performed June 4 to 6, 1984)
Sequlm Bay
Case Inlet
Station
141
17
18
20
Control
No.
Rep. Scored
h
B
C
D
Total
A
B
C
D
Total
A
B
C
D
Total
A
B
C
D
Total
A
B
C
D
51
50
124
31
256
43
49
10
89
191
87
45
59
55
246
32
63
67
53
215
255
268
241
251
No.
Abnormal
9
0
3
0
12
1
8
1
7
17
3
12
10
13
38
2
6
5
9
22
12
31
13
13
Percent
Abnormal
4.7
8.9
15.4
10.2
No.
Station Rep. Scored
1 A
B
C
D
11
15
17
Control
Total
A
B
C
D
Total
A
B
C
D
Total
A
B
C
D
Total
E
F
G
H
57
73
71
119
320
10
11
56
52
129
61
93
83
80
317
60
62
28
51
201
261
289
282
245
No.
Abnormal
6
8
16
25
55
1
1
8
7
17
5
13
4
7
29
4
5
2
2
13
15
29
37
20
Percent
Abnormal
17.2
13.2
9.1
6.5
Total 2092
170
8.12
1 Sediment control.
2 Percent abnormal of 8 control test replicates.
-------
TABLE 57. Summary Analysis of Variance for Percent Abnormality
(Arcsin /X) in Each Oyster Larval Bioassay
Source of
Variation
Degrees of
Freedom
Sum of Squares
Mean
Square F-Ratio
Assay No. 1
Mean
Stations
Error
Total (corr)
Mean
Stations
Error
Total (corr)
Mean
Stations
Error
Total (corr)
Mean
Stations
Error
Total (corr)
1
8
30
38
1
8
26
34
1
8
30
38
1
8
31
39
1.323
0.817
2.299
3.116
Assay No. 2
1.538
2.469
0.565
3.034
Assay No. 3
3.304
0.417
1.118
0.536
Assay No. 4
1.226
0.231
0.091
3.321
0.102 1.331
0.077
0.309 14.213
0.022
0.052 1.401
0.037
0.029 9.893
0.003
i Not significant.
2 Significant (p <0.05).
3 Significant (p <0.01).
175
-------
TABLE 57. Summary Analysis of Variance for Percent Abnormality
(Arcsin S%) in Each Oyster Larval Bioassay
(Continued)
Source of
Variation
Mean
Stations
Error
Total (corr)
Mean
Stations
Error
Total (corr)
Degrees of
Freedom
-
1
8
31
39
1
8
31
39
Sum of Squares
Assay No. 5
3.144
0.392
0.235
0.627
Assay No. 6
3.717
0.215
0.325
0.540
Mean
Square
0.049
0.008
0.027
0.010
F-Ratio
6.453
2.562
176
-------
TABLE 58. Comparison of Treatment and Control Abnormality
(Arcsin /X) Using Dunnett's Test
Bay
Station
Mean
Bay
Station
Mean
Bay
Station
Mean
Bay
Station
Mean
Bay
Station
Mean
Bay
Station
Mean
Samish
Assay No. 1
Bellingham
I 3 7 2lT 3^ A 5 7
.153 .044 .169 .118 .113 .107 .161 .029
Assay No. 2
Port Gardner -
Bellingham Everett Harbor
11 12 23 2T 12^^
.3441 .3451 1.571 .3581 .147 .000 .123 .069
Assay No. 3
Port Gardner -
Everett Harbor
Fourmile Rock-
Elliott Bay2
b 6 7 U ^10 12 IT
.524 .217 .271 .263 .266 .141 .354 .399
Assay No. 4
Fourmile Rock-
Elliott Bay2
Sinclair
2(T 22 23 2A 6 7 8^ U
.115 .060 .210 .130 .2611 .2711 .185 .2911
Assay No. 5
Sinclair
Da bob
IT 18 19 20 I 5/15
.2901 .259 .4861 .3501 .3151 .242 .3251 .278
Assay No. 6
1 Significant (p <0.05).
2 Dump site vicinity.
Control
.457
Control
.074
Control
.219
Control
.114
Control
.130
Sequim
14
.147
17
.294
18
.415
20
.317
8
.410
Case
11
.348
15
.299
17
.255
Control
.282
177
-------
TABLE 59. Significance of Oyster Larval Percent Abnormality
Data by Bay and Station for Sediments Collected
During Detailed Surveys April 23 to May 29, 1984
Location Significant1 Not Significant
Samish 1, 3, 7, 20
Dabob 1, 7 5, 15
Sequim 14, 17, 18, 20
Case 8, 11, 15, 17
Bellingham 11, 12, 23, 24 3, 4, 5, 7
Four-mile Rock-Elliott Bay2 9, 10, 12, 17
20, 22, 23, 24
Port Gardner-Everett Harbor 1, 2, 3, 4
5, 6, 7, 11
Sinclair 6, 7, 14 8, 18
17, 19, 20
1 Differed significantly from Control (p <0.05).
2 Dump site vicinity.
178
-------
survival. The trend to higher percentages of abnormal oyster larvae
becomes more evident as amphipod survival decreases.
Salinity, pH, and dissolved oxygen values remained at acceptable
levels in all cultures during the assays. Control cultures also generally
showed low percentages of abnormal larvae: Assay No. 1, 4.2%; Assay
No. 2, 0.72; Assay No. 3, 4.22; Assay No. 4, 1.52; Assay No. 5, 1.9%; and
Assay No. 6, 8.1%. Although the percentages of abnormal larvae exceeded
the 3% abnormal rate suggested by Woelke (1972) as acceptable for controls
without correction in only three of the assays, we chose to correct all of
the data (Appendix E). The ASTM (1980) suggests that if abnormality
exceeds 2% in control larvae, treatment abnormality should be corrected
using Abbott's equation (Finney 1980). It should also be noted that ASTM
(1980) allows use of control abnormalities of up to 10% provided the data
are corrected and weighted mean control abnormality values are reported.
Survival data were not routinely reported. We focused on abnormal
shell development because it is typically more sensitive (ASTM 1980). It
is also a less variable parameter and, hence, is a more reliable endpoint
for use in bioassays.
Assay No. 5 was rescored by again subsampling the remaining screened
larvae from each test container. In this case, the first 200 larvae were
evaluated. This procedure was necessary because many of the subsamples
used for initial scoring contained too few larvae for meaningful
statistical comparisons. It was likely that the larvae were not uniformly
distributed within the test cultures at the time of subsampling. Either
the cultures were not thoroughly mixed, or larvae were "bound" to
particulates that rapidly settled to the bottom of test containers. The
problem was also encountered in a few test containers in Assay Nos. 1, 2,
3, and 6, but was decidedly more pronounced in Assay No. 5.
The inclusion of six oyster larval bioassays on screening survey
sediments served to calibrate our methods and, in particular, to test
various modicications of ASTM E 724-80 (ASTM 1980) to adapt the technique
to sediment testing. First, these data suggested that the test sediments
be added at least to the 10 or 100 g/L concentration level. Various other
179
-------
investigators (Schink et al. 1974; Cummins et al. 1976) have used sediment
concentrations from 0.5 to LO g/L, and different methods for exposure
including rotating the embryos together with test sediments (Cardwell
et al. 1977). Although the method described here used higher sediment
concentrations than others have reported, there appeared to be little or
no physical effect using our method of exposure. Second, these data
suggested that perhaps rigorous use of a sediment control was not
required. The addition of a clean sediment from Sequim Bay (Station 14)
over a range of sediment concentrations of 0.01 to 100 g/L failed to evoke
a response different from the seawater controls. Accordingly, while we
did not elect to include a sediment control in each batch of bioassays
performed during the detailed surveys (see Section 3.5.2), we did provide
for Sequim Bay Station No. 14 to be retested as part of the detailed
surveys. It also seemed inappropriate to use other types of sediment
controls such as a No. 4 washed mortar sand, or a manufactured sand
specifying grain size, percent water, and certain chemical factors.
Although Chapman and Morgan (1983) used a sediment control, they did not
specify source or physical or chemical constituency.
3.7 SUMMARY STATISTICS
Summary scores for chemical, biological, and physical variables used
in the statistical exercise to determine which bays were showing signs of
degraded sediment quality are presented in Table 60.
The mean of the summary metals scores from the 16 baseline stations
was 0.0516 with a standard deviation of 0.0150. The tolerance factor, K,
for this distribution to give a probability of 0.99 that 99% of the
distribution lies within x ± Ks is K = 4.492. The upper limit for this
tolerance interval is 0.1191. A score of 0.120 or greater was considered
unlikely to be attained in an unimpacted environment.
The mean of the summary organics scores from the baseline bays was
0.0126 with a standard deviation of 0.0028. The tolerance factor, K, is
the same, 4.492. The upper tolerance limit of the distribution is 0.0250.
180
-------
A score of 0.026 or greater was considered indicative of an impacted
environment.
The critical level of 0.20 for proportion of dead amphipods was
established in Section 3.5. The stations that were found to be different
from controls in the oyster larvae bioassay were identified in
Section 3.6. All summary scores exceeding their appropriate critical
value are indicated by (1). The column designated "signal" in Table 60
denoted whether a chemical, or biological variable, or both variables
exceeded the critical value.
On the basis of the data presented in Table 60, each of the
48 stations can be classified as exhibiting "signals" in one of four
categories:
1. no biological or chemical signal
2. a biological, but no chemical signal
3. a chemical, but no biological signal
4. both a chemical and a biological signal.
All stations from the urban bays provided at least a chemical signal.
The Fourmile Rock - Elliott Bay dump site vicinity showed only chemical
signals at all stations. The other three urban bays indicated both
chemical and biological signals at all or almost all of their stations.
Port Gardner - Everett Harbor and Sinclair Inlet demonstrated the most
consistent and strongest biological and chemical signals.
Among the baseline bays, all stations in Case Inlet and Samish Bay
showed biological signals (lowered amphipod survival) in the absence of
chemical signals. Sequim Bay provided the greatest proportion of stations
without any signal.
181
-------
TABLE 60. Summary Scores for Chemical, Biological, and
Physical Variables Used to Determine S.igns
of Degraded Sediment Quality
Bay
Oyster
Station Meta 1 s Orgam'cs Amphipods Larvae Capital la Fines
Signal'
Sami sh
Dabob
Sequim
Case
Bel 1 i ngham
Fourmile Rock-
Elliott Bay2
1
3
7
20
1
5
7
15
14
17
18
20
1
11
15
17
3
4
5
7
11
12
23
24
9
10
12
17
20
22
23
24
0.041
0.052
0.042
0.047
0.026
0.029
0.041
0.062
0.047
0.058
0.057
0.054
0.084
0.049
0.072
0.065
0.1711
0.1211
0.096
0.102
0.089
0.072
0.090
0.083
0.2501
0.5311
0.107
0.1621
0.2631
0.117
0.053
0.056
0.016
0.016
0.014
0.014
0.008
0.008
0.009
0.012
0.012
0.012
0.016
0.012
0.017
0.011
0.014
0.012
0.1861
0.2071
0.0761
0.1521
0.0661
0.0721
0.0821
0.0751
0.1461
0.2851
0.0401
0.1621
0.2691
0.2091
0.0681
0.0581
0.3101
0.4701
0.2501
0.3101
0.100
0.070
0.2601
0.3601
0.120
0.100
0.110
0.2701
0.2801
0.2801
0.4301
0.4101
0.040
0.170
0.3401
0.050
0.170
0.110
0.5801
0.110
0.130
0.140
0.160
0.080
0.170
0.140
0.120
0.130
0.000
0.000
0.000
0.000
0.0841
0.029
0.0931
0.061
0.000
0.009
0.079
0.023
0.099
0.055
0.011
0.000
0.000
0.000
0.000
0.000
0.0881
0.1471
1.0001
0.0981
0.035.
0.009
0.139
0.254
0.000
0.000
0.016
0.003
0
0
0
0
0
1
0
0
0
0
0
0
0
0
0
0
0
91
0
0
0
0
1
0
0
1
0
0
0
0
0
1
0.812
0.809
0.849
0.873
0.203
0.237
0.490
0.897
0.639
0.831
0.794
0.839
0.894
0.390
0.813
0.781
0.669
0.836
0.965
0.917
0.981
0.642
0.954
0.974
0.557
0.689
0.886
0.559
0.523
0.624
0.245
0.137
bio
bio
bio
bio
bio
none
bio
bio
none
none
none
bio
bio
bio
bio
bio
chem
bio-chem
bio-chem
chem
bio-chem
bio-chem
bio-chem
bio-chem
chem
chem
chem
chem
chem
chem
chem
chem
1
Higher than critical value.
Dump site vicinity.
Signs of degraded sediment quality based on chemical,
biological, or both a chemical and biological variable
exceeding critical value.
182
-------
TABLE 60. Summary Scores for Chemical, Biological, and
Physical Variables Used to Determine Signs
of Degraded Sediment Quality (Continued)
Bay
Oyster
Station Metals Orgam'cs Amphipods Larvae Capitella Fi nes
Signal'
Port Gardner- 1
Everett Harbor 2
3
4
5
6
7
11
Sinclair 6
7
8
14
17
18
19
20
0.1341
0.087
0.1301
0.2701
0.118
0.060
0.088
0.071
0.3731
0.6701
0.3091
0.3341
0.3261
0.2441
0.4341
0.3441
0.6031
0.2031
0.4461
0.7681
0.2941
0.1081
0.1601
0.0611
0.3871
0.2961
0.2021
0.4581
0.2361
0.1761
0.2651
0.7341
0.5601
0.2901
0.3301
0.5801
0.8601
0.110
0.100
0.2601
0.4501
0.190
0.3801
0.4301
0.2601
0.3201
0.190
0.3801
0.022
0.007
0.009
0.009
0.225
0.014
0.023
0.002
0.0501
0.0531
0.020
0.0531
0.0681
0.051
0.2081
0.1021
161
171
11 1
W1
50 1
61
1
0
0
0
0
0
0
0
0
0
0.629
0.831
0.635
0.538
0.673
0.402
0.675
0.691
0.868
0.555
0.899
0.916
0.782
0.778
0.490
0.686
^—^—*^^^~^— —
bio-chem
bio-chem
bio-chem
bio-chem
bio-chem
bio-chem
chem
bio-chem
bio-chem
bio-chem
bio-chem
bio-chem
bio-chem
bio-chem
bio-chem
bio-chem
Higher than critical value.
Dump site vicinity.
Signs of degraded sediment quality based on chemical,
biological, or both a chemical and biological variable
exceeding critical value.
183
-------
Page Intentionally Blank
-------
4.0 DISCUSSION
4.1 CONTAMINANT LEVELS
4.1.1 Relationship to Physical Properties of Sediments
Examination of the correlation matrix in Table E.2 revealed that
average phi was strongly correlated (p <0.001) with water content.
Also, percent volatile solids correlated (p <0.001) with water content.
An unexpected finding was the lack of significant correlations between
average phi and contaminants, either metals or organics. Previous
geochemical studies in baseline bays and in the main basin of Puget Sound
reported strong correlations between grain size and contaminant
concentration (Crecelius et al. 1975; Dexter et al. 1981; Romberg et al.
1984). The apparent reason for the lack of correlation in our study was
the finding that some coarse-grain sediments from the shallow urban bays
were heavily contaminated with both metals and organics. Coarse-grained
sediments are not usually highly contaminated unless they occur near a
point source of pollution as in the case of some stations in urban bays.
A strong correlation (p <0.05) among many heavy metals in sediments
was observed. In urban bays, heavy metals often have common sources and
are associated with the same suspended particles. Therefore, sediments
are usually contaminated by a group of heavy metals.
The organic chemicals and organic properties of the sediment were
fairly strongly correlated. The correlation between TOC and percent
volatile solids was very high (p <0.001) as was expected because most of
the volatile matter in sediment is organic matter. PCB-1254 was also
highly correlated (p <0.01) with aromatic hydrocarbons. PCBs were highly
correlated (p <0.001) with Ag perhaps because the major source of both
contaminants is sewage. Aromatic hydrocarbons were highly correlated
(p <0.001) with IR, as were PCBs (p <0.01).
185
-------
4.1.2 Comparison of Contaminants in Urban and Baseline Bays
In this study, concentrations of metals, PCB-1254, and aromatic
hydrocarbons in urban bays were clearly different from those in baseline
bays. Urban bay sediments were generally clearly contaminated by Ag, As,
Cu, Cd, Hg, Pb, and Zn. Other metals (Sb, Be, Cr, Ni, Se, Ti) were found
at about the same concentration in all bays, however, a few urban bay
sediments were enriched in Sb, Cr, and Ni. Baseline bays generally
contained lower concentrations of metals.
The only organics that were consistently quantified in urban bays
were the aromatic hydrocarbons and PCB-1254. Other compounds occasionally
quantified in urban bays included phthalates and PCB-1260. By comparison,
baseline bay sediments generally contained less organic contamination. No
PCBs were detected, and only a few stations in one baseline bay, Samish
Bay, were found to contain detectable amounts of phthalates and aromatic
hydrocarbons.
It is tempting to define baseline or "reference" conditions on the
basis of chemical variables. Sequim Bay and Dabob Bay, with relatively
low metals burdens and no detectable organic compounds, appear to be
excellent choices for reference bays. Application of this concept is
potentially useful in a regulatory sense to determine where and when
remedial action is required.
The fact that all bays in Puget Sound interconnect and that
circulation transports contamination from one bay to another, however,
tends to detract from the concept of "reference" bays. Circulation models
developed for Puget Sound predict that gradients of both dissolved and
particulate materials exist between regions of higher and lower
contamination (Ebbesmeyer et al. 1984).
4.1.3 Comparisons to Other Studies
The range of contaminant concentrations in surface sediments of the
urban and baseline bays were compared with mean data from other sediments
in Puget Sound (Table 61), including pre-1900 sediments. The urban
central basin sediments (Area BB in the METRO TPPS Report [Romberg et al.
186
-------
TABLE 61. Comparisons of Concentrations of Contaminants in Puget Sound Sediments
IM» Stwto
».H»» «t «l. 1MB 4 1»
Niln Bwln Rock-
Port CardMf- Surf.c. Elllottftiy
Ey.r.tt Itorbor Swll««itl Crlt.rl*
So. Sound
Pugtt Pr«-t90C
Sound* S«dl««nt2
FouralU FovralU
BolllnghM Rock- Port CardMr- Slnclilr BolllnghM Rock- Port Ctrdrur- SlncUIr Co«encMent
B«y Elliott B« tv«r«tt Hjrbor lnl«t R«na* B
100 110
1C-22
57
IO-»»
00
Hg
(PP.)
Cu
(PP>)
AH
(PP>)
PCB
1.0
110
15.9
1.5
».J
O.J» 2.1
».J J19
O.OC- l.»
0.1}
J7-»9 t*f>
0.15
579 <20 <100
O.M 0.1C 0.91 0.3C 0.07-
0.»7
£8 NO*! 127 199 10-U
1i 2.7 «.» 9.7 0.25-
0.»8
0.5}
1JO 270
J-10 400
O.U
>C
2.2
125
O.OC O.M
0.1}
21-7C 21
0.1- 0.2*
O.B
OO 9.1
1 Intorg «t «1. 19M.
2 M1«y «t «1. 1M>.
3 And«r>on «td Cr«c«llu« 1915.
" Not doUraliMd.
-------
1984]) are representative of fine-grain sediments now accumulating in the
main basin of Puget Sound. The Fourmile Rock - Elliott Bay data have been
used by U.S. EPA Region 10 to set Interim Decision Criteria for open water
disposal at Fourmile Rock in Puget Sound.
Several elements in baseline bays, including Sb, As, Be, Cd, Cr, Ni,
Se, Tl, and Zn, were found at approximately the same concentrations as in
the pre-1900 sediments. This similarity indicated that anthropogenic
activities have not contaminated the baseline bays with these nine
elements. Current baseline concentrations of Sb, Be, Se, and Tl were less
than those measured in pre-1900 sediments probably because analytical
techniques and detection limits were different. Several elements,
including Be, Se, and Tl were not elevated even in urban sediments.
The elements As, Cd, Cr, Ni, and Zn were generally elevated by not
more than a factor of 10 in the urban bays compared to the baseline bays
and baseline sediments deposited in the 1800s (Table 62).
In some urban sediments, metal concentrations were more than 50 times
greater than concentrations in baseline sediments. These metals included
Cu, Pb, Hg, and Ag. The concentrations of these metals were also enriched
in many of the baseline bay sediments. Possible sources of metals
entering baseline bays include transport of contaminants from the main
basin of Puget Sound, local point and nonpoint sources, and atmospheric
fallout. For example, Case Inlet, which was the most contaminated
baseline bay, has been shown to act as a sediment trap of contaminants
transported from the Tacoma area into the southern Sound (Riley et al.
1983).
We selected three metals (Pb, Hg, and Cu) and two classes of organic
compounds (aromatic hydrocompounds and PCBs) to compare contaminants in
sediments of the urban and baseline bays with other bays or regions of
Puget Sound. These five contaminants were chosen because they are
frequently quantified in sediments and are almost always elevated in
sediments from urban areas. Also, these contaminants are on the Fourmile
Rock Site Interim Decision Criteria list. Table 61 lists the mean
concentrations of the five contaminants in the four urban bays and
188
-------
As already noted, disturbed stations with major shifts in dominant
infauna only occurred in urban bays (Bellingham Bay and Port Gardner -
Everett Harbor). These stations accounted for the most dramatic
differences between urban and baseline bays.
After omitting the stations with high TOC levels and comparing the
averages for urban and baseline bays within each sediment category
(Table 22), other differences in infaunal patterns between urban and
baseline bays became evident. The number of taxa (richness) in silty-mud
of the urban bays was half that in the same sediment type of baseline
bays. For stations with sandy-silt, richness was lower in the urban bays
when compared to baseline bays. For the sandy stations, richness was
about the same in both urban and baseline bays. Also, in silty-mud, the
average abundance was the same for urban and baseline bays. In contrast,
for sandy-silt, the average abundance in urban bays was 40% of that in
baseline bays. However, this latter difference was confounded by
differences in depth. The Fourmile Rock - Elliott Bay dump site vicinity
had five deep stations in the sandy-silt group; and these deep sandy-silt
stations showed low abundance. The sandy-silt stations in Sinclair Inlet
were shallower than those in baseline bays and demonstrated intermediate
to high abundance in the same range as those in Sequim Bay, the most
comparable baseline bay. Setting aside the influence of high TOC levels,
distinct species compositions for urban bays were not distinguished. For
stations with silty-mud, the most abundant species in both urban and
baseline bays were the Cirratulid polychaetes, primarily Tharyx spp. For
stations with sandy-silt, Axinopsida serricata became the most abundant
species in some urban bays. However, this shift to Axinopsida serricata
occurred at the deeper stations with sand contents above 30% and was
probably because of habitat differences rather than being from an urban
bay. For silty-sand and sand, the shift from Axinopsida serricata to
Cirratulid polychaetes at one urban station was probably related more to
the shallow depth of the bay rather than to the type of bay.
191
-------
4.2.3 Comparisons With Other Studies
Overall, the results of the benthic infauna analyses confirm the
general findings of other studies of Puget Sound infauna. Lie (1968) had
already shown that substrate type was a dominant factor in determining the
character of benthic infauna in Puget Sound. Our finding of a change in
species composition from mud to sand reinforces Lie's general conclusion.
Similarly, Nichols' (1970) general conclusion that clay content of the
sediment controls polychaete distribution is consistent with our finding
that high clay content was associated with sparse infaunas. The general
conclusions of these two studies strengthen our belief that the
development of sediment quality criteria must entail examination of the
effects of contaminants for different sediment types.
Most infaunal studies in Puget Sound have focused on sewage outfalls.
Consequently, there is little overlap between our stations and those of
other studies. Consistent with our observations is the observation by
others of large abundances of enrichment opportunistic species in areas of
high organic content in Puget Sound. For example, Armstrong et al. (1980)
reports a shift to high abundance of Capitella capitata around a combined
sewer overflow at 9 m depth in Elliott Bay. The Fourmile Rock - Elliott
Bay dump site vicinity stations sampled in our study are much deeper (over
100 m) than those of Armstrong et al. (1980).
The recent study of Swartz et al. (1985) along a gradient of stations
in relation to a California marine sewage outfall provides two major
findings that are similar to those in this study. First, the three
stations closest to the sewage outfall had the highest organic content
(TOC>4.0%) and revealed infaunas dominated by Capitella capitata.
Second, the same three stations where Capitella capitata dominated also
had significantly lower amphipod survival. In addition, these impacted
stations showed lower species richness and abundance. These findings of
Swartz et al. (1985) compare well with those of this study and strengthen
the general expectation that organic enrichment of sediment leads to a
shift toward infauna dominated by particular opportunistic species and to
sediment toxicity for amphipods.
192
-------
In another study, Chapman et al. (1984a, b) applied several types of
multivariate analyses to chemical and biological data from 23 stations in
four urban Puget Sound bodies of water, Elliott Bay (5 stations), the
Duwamish Waterway (3 stations), Commencement Bay (10 stations), and
Sinclair Inlet (2 stations), as well as two nonurban bays, Case
Inlet (1 station) and Samish Bay (2 stations). Their Elliott Bay stations
were shallower (less than 40 m) than ours and located closer to shore in
proximity to sewage outfalls. Factor and cluster analyses applied to
benthic infauna data alone and to infaunal and chemical data combined
showed that Case Inlet and Samish Bay were distinct from the urban bays.
As in our study, the Samish Bay infauna revealed an important echinoderm
component virtually absent from the urban bays and demonstrated a species
composition distinct from the other bays. Based on our results, the
finding that two nonurban bays (Sequim Bay, Dabob Bay) showed infaunas of
the same general species composition as that of the urban bays (Sinclair
Inlet, Bellingham Bay, some stations in Port Gardner - Everett Harbor)
raises the question of whether just two stations from Samish Bay and one
from Case Inlet constituted an adequate and appropriate control or
baseline for multivariate analyses.
Earlier studies (1964-1966) in Bellingham Bay and Everett Harbor have
reported stations with extremely impoverished infauna where sludge from
pulp mill effluent had accumulated (Jones and Stokes Associates 1983).
More recent studies in Everett Harbor found some recovery but the infauna
near the pulp mill outfalls was still dominated by Capitella capitata
(Jones and Stokes Associates 1983). Our results indicate that areas with
organically enriched sediment still exist in Everett Harbor and to a
lesser extent, in Bellingham Bay, and that such areas are still dominated
by organic enrichment opportunists.
193
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4.3 FISH AND SHELLFISH PATHOLOGY
4.3.1 Relationships Between Lesion Incidence and Sediment Chemical
Composition
4.3.1.1 Fish
Hepatic Lesions. Two major lines of evidence suggest that three of
the hepatic lesions in fish (neoplasms, preneoplasms, and specific
degeneration) are causally related to exposures of environmental
contaminants. First, in studies previously conducted in Puget Sound
(Maiins et al. 1982, 1984; McCain et al. 1982), these types of hepatic
lesions were found only, or were significantly more prevalent, in flatfish
from chemically contaminated urban embayments. The urban areas with the
highest lesion incidences were the Duwamish Waterway and the downtown
waterfront in Seattle, the waterways of Commencement Bay, and Everett
Harbor. Second, these types of lesions have been reported to occur in
laboratory animals exposed to toxic and/or carcinogenic chemicals.
Lesions morphologically similar to specific degeneration have been induced
by PCBs in mammals and fish (Koller and Zinkl 1973; Hinton et al. 1978).
Hepatic neoplasms and preneoplasia are inducible in mammals by many
chemicals, including carbazole and derivatives (Tsuda et al. 1982), DDT,
dieldrin, N-acetyl-2-aminofluorene, nitrosamines and phenabarbitone
(Jones and Butler 1975). A variety of elements, including arsenic,
chromium, silver, indium, iron, molybdenum, selenium, tellurium and
thallium also possess hepatotoxic properties in humans and mammals
(Beliles 1975). In fish, heptocellular and cholangiocellular neoplasms
similar to those seen in our study have been produced by exposure
to N-methyl-N'-nigrosoguanidine (Hendricks et al. 1980), and
7,12-dimethylbenz[a]anthracene (Schultz and Schultz 1982).
The other types of hepatic lesions, storage disorders and nonspecific
degenerative lesions, were found in both urban and baseline bays, although
highest incidences were generally found in fish from urban bays (Maiins
et al. 1982, 1984). The remaining three types of hepatic lesions—
194
-------
vascular, proliferative, and inflammatory disorders—were evenly
distributed in both urban and baseline areas. All five types of these
lesions tend to be nonspecific responses to a variety of factors,
including exposure to natural and synthetic toxins and physiological
changes related to reproduction or diet.
Significance of Kidney and Gill Lesions. Relationships between
kidney and gill lesions in fish and chemical contaminants are not as
strong as those observed for hepatic lesions. Necrosis and certain types
of depositional disorders of the kidney have been found in English sole
from both urban and nonurban areas of Puget Sound. However, the highest
incidence of the kidney necrosis was found in the Hylebos and Duwamish
Waterways (Malins et al. 1982), and the depositional disorders were most
prevalent in the Duwamish Waterway and the estuary of the Lake Washington
Ship Canal in Seattle (McCain et al. 1982).
A similar distribution pattern in both urban and nonurban areas was
observed for the proliferative gill conditions in English sole. Malins
et al. (1982) found the highest incidence in the Hylebos Waterway and
Discovery Bay, and McCain et al. (1982) found such conditions only in sole
from the Duwamish Waterway, Lake Washington Ship Canal, and at the mouth
of the Snohomish River in Everett. Proliferative gill lesions have been
found in sole widely distributed throughout Puget Sound, and can be
caused by external parasitism as well as exposure to chemical irritants.
Consequently, their significance in relation to sediment pollutants is
questionable.
4.3.1.2 Shellfish
Vascular Disorders. These conditions were found only in the gills of
the shrimp species, and were more prevalent in shrimp from the baseline
bay, Dabob Bay. In several instances, congestion of the gill filaments
was associated with hyalinocytic phagocytosis and encapsulation. The gill
has a prime function in the elimination of foreign and necrotic material
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(Fontaine and Lightner 1975). The congestion, particularly in association
with phagocytosis and encapsulation, may represent efforts by the affected
shrimp to remove parasites, cellular debris, or foreign materials from the
animal, and, thus, does appear to be a serious condition. The cause of
gill microaneurysms is largely unknown, but may be an artifactual finding
related to pressure changes involved in capturing the animals at depth and
then bringing them too quickly to the surface. These lesions were not
associated with any observable parasite, and they occurred in all sampling
areas.
Inflammatory Conditions. Hyalinocytic infiltration, encapsulation,
and phagocytosis were also detected only in shrimp species, and except for
encapsulation in the antennal gland and phagocytosis in the gill, were
more prevalent in shrimp from Dabob Bay. The function of these processes
in shrimp is generally to isolate and/or remove foreign and necrotic
material, whether caused by parasites, physical debris, or chemical
destruction of cells (Couch 1978; Doughtie and Rao 1983; Fontaine and
Lightner 1975; Johnson 1980). Much of the encapsulation in the
hepatopancreas was associated with a parasitic infection in £. dispar,
which surrounded melanized lesions in the connective tissue between the
digestive tubules. Hemocytic (hyalinocytic) infiltration accompanied
these occasionally severe infestations, and hyalinocytes were observed
phagocytosing small dense particles. Even in severe cases, the digestive
tubules appeared quite normal, except that no B-cells (Blasenzellen) were
present in the digestive epithelium. No discernible parasites could be
detected in association with inflammatory conditions in the hepatopancreas
of £_. platyceros from Dabob Bay.
The incidence of hemocytic infiltration in the antennal gland was
highest in C_. magister from Bellingham Bay; this condition in the eye was
equally prevalent in all three sampling areas. Hemocytic infiltration of
the epithelium of the esophagus and ommatidium of the eye were minor and
did not appear to represent a serious impairment. Infiltration of the
coelomoduct of the antennal gland represents a potentially more serious
disorder. However, the infiltration was minor in all affected animals,
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and the disorder was found in crab (both C_. magister and C. gracilis) from
all areas except Dungeness Spit.
Melanized lesions and unmelanized granulomas can develop for a
variety of reasons. Injury and disease cause the majority of these
responses (Bang 1983; Fontaine and Lightner 1975; Lightner and Redman
1977). Exposure to a variety of chemicals can also cause melam'zation
reactions (Couch 1977; Doughtie and Rao 1983, 1984; Ninrno et al. 1977).
In addition, dietary deficiencies can also cause these lesions (Lightner
et al. 1977; Smith and Taylor 1968). Melanized lesions and unmelanized
granulomas in crab, and melanized lesions in shrimp all tended to be more
prevalent in tissues of animals from baseline bays than in urban bays.
This evidence would seem to preclude environmental pollutants in urban
bays as a probable cause for these lesions. Parasitic infections tended
to be more prevalent, at least in £. dispar and C. gracilis, in baseline
bays. The presence of undetectable (at the light microscope level)
parasites in the other crab and shrimp species may also have been
associated with the higher incidence of melanized lesions and unmelanized
granulomas in reference areas.
Degenerative Disorders. Although the incidence of degenerative
disorders in both crab and shrimp was low, these lesions in the antennal
gland of £. dispar and £. platyceros, and in the heart of £. platyceros,
were the only ones that appeared to be exclusively associated with the
Fourmile Rock - Elliott Bay dump site vicinity. In the heart of
£. platyceros, the degenerative changes were associated with a suspected
parasitic infection.
Hydropic degeneration/membrane lysis occurred only in the
hepatopancreas of C. magister while nuclear atypia occurred in the
hepatopancreas of both crab species. The high concentration of digestive
enzymes present in the epithelial cells of the hepatopancreas can result
in rapid dissolution of the plasma membrane and cause autolytic changes in
the epithelial cells when the crab are subjected to the stress of handling
and necropsy. The fact that hydropic degeneration/membrane lysis occurred
only in C_. magister probably represents a more delicate nature of the
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digestive epithelium of this species. This condition was also much higher
in C. magister from the inner waterways of Bellingham Bay (Stations 11 and
12 and Stations 4 and 5), and was not observed in crab from Dungeness
Spit. The digestive epithelium of other hepatopancreatic tubules did not
appear to be affected by this condition. The gradual decrease in
incidence of this condition from inner areas to outer areas of Bellingham
Bay (Stations 23 and 24) and Samish Bay, and the total absence of this
condition in crab from Dungeness Spit, suggest that perhaps subtle
degenerative changes may affect membrane stability occurring in the
hepatopancreatic epithelial cells of crab from the inner areas of
Bellingham Bay. These changes may predispose the hepatopancreas to
membrane lysis when the animals are exposed to stress.
Nuclear atypia may also be the result of autolytic changes resulting
from stress of capture, handling, or necropsy and appeared in both crab
species. However, the condition was present relatively rarely within the
hepatopancreas and was observed in crab from all sampling areas.
Proliferative Conditions. Proliferative conditions as represented by
an increase in mitotic activity in various tissues can result from a
number of factors: the stage of the molting cycle, feeding activity,
parasitic infection, and environmental or physical stress (Johnson 1980).
No distinct pattern of the incidence of proliferative conditions was
detected in C. gracill's. The significantly higher incidence of these
conditions in the hemopoietic tissue in £. magister from Stations 11 and
12 in Bellingham Bay, compared to the significantly lower incidence of
this lesion in crab from Samish Bay, may represent a reaction to chemical
injury at this site. However, the possible relationship to chemically
contaminated environments is unclear because the incidence of this lesion
in crab from Dungeness Spit was similar to that of two sites in Bellingham
Bay.
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4.3.2 Comparison of Fish and Shellfish Pathology in Urban and Baseline
Bays
Although the original design of this study called for collecting
flatfish from eight embayments in or near Puget Sound, flatfish were not
collected from four of these areas (Bellingham Bay, Sequim Bay, Samish
Bay, and Dabob Bay) because adult English and Dover sole (fish >150 mm in
length) were scarce or absent. Three of these baseline bays (Samish Bay,
Dabob Bay, and Sequim Bay) were intended to serve as "reference" areas.
To partially compensate for this problem, an additional baseline station
near Eliza Island (west of Bellingham Bay, Station No. 04015) was sampled,
and sufficient numbers of adult English sole were collected. Mai ins
et al. (1982) demonstrated that English sole less than two years old had
significantly (p <_0.05) lower incidences of idiopathic liver diseases than
fish over two years old. For example, liver neoplasms were not detected
in males less than two years old or in females less than one year old. No
appropriate baseline station was found for Dover sole. To correct for
this situation, stations outside of, but adjacent to, the Fourmile Rock -
Elliott Bay dump site vicinity were sampled. Of these two stations (one
southeast of West Point [Station No. 10065] and the other near Duwamish
Head [Station No. 10064]), sufficient numbers of adult Dover sole were
captured only at the West Point Station.
This difficulty did not exist for shellfish as suitable numbers of
crab and shrimp were collected from all sampling areas except Sequim Bay.
Cancer magister collected from Dungeness Bay were substituted for Sequim
Bay crab.
4.3.2.1 Urban Bays
Fish. For English sole, Sinclair Inlet had the highest incidences of
liver, kidney, and gill lesions of all the urban bays sampled. Within the
Inlet, the incidence of lesions tended to be higher near the northwestern
portion. For Dover sole, relatively low percentages of occurrence of
serious liver lesions were detected in the Fourmile Rock - Elliott Bay
dump site vicinity near West Point. Although these observations with
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Dover sole provide only suggestive information about possible effects of
environmental contaminants in these two areas, they do demonstrate that
Dover sole may serve as a very useful indicator fish species for
assessment of environmental stresses in the deeper portions of Puget
Sound.
Shellfish. A number of pathological conditions in shellfish were
also identified in the urban bays. The relationships of many of these
conditions to urban-associated contamination are poorly understood at this
time. Nevertheless, two types of lesions were found only in animals from
urban areas, suggesting that they were caused by urban-associated factors
(e.g., chemical contamination). Hydropic degeneration/membrane lysis in
the hepatopancreas of C. magister and degenerative disorders in the
antennal gland of both shrimp species were observed in Bellingham Bay and
in the Fourmile Rock - Elliott Bay dump site vicinity, respectively.
4.3.2.2 Baseline Bays
Fish. Of the four baseline bays selected for this study, only Case
Inlet had sufficient numbers of adult sole to serve as a baseline bay.
The Eliza Island baseline station near Bellingham Bay was added to
partially correct for this situation. Although the incidence of serious
liver, kidney, and gill lesions in English sole from Case Inlet was low
compared to incidences previously detected in sole from highly
contaminated urban areas (e.g., the Duwamish Waterway), these lesions were
not detected in sole from the Eliza Island station. It is currently not
known whether the lesions observed in sole from Case Inlet were induced in
situ or were in fish that migrated there from a contaminated area, such as
Commencement Bay.
Shellfish. Most identified pathological conditions were found to be
more prevalent in the baseline bays for both crab and shrimp species.
These conditions are believed to have been the result of existing or
previous parasitic infections.
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In both shrimp species, proliferative conditions occurred in several
tissues and in general were more prevalent in animals from Dabob Bay. The
most probable cause of this condition would be parasitic infection. For
example, in the hepatopancreas of P_. dispar the higher incidence of
proliferative conditions was accompanied by an identifiable parasitic
infection. However, from the evidence available, proliferative conditions
could not be attributed to any particular causative agent. In the case of
shrimp, the general trend toward greater incidence of proliferative
conditions in baseline areas would seem to rule out pollution in the
etiology of this condition.
Parasitic/Infectious Disorders. Parasitic infections in crab and
shrimp species were detected in both urban and baseline areas. The higher
incidence of parasitic infections in crab and shrimp from baseline areas
does not permit any conclusions as to whether crab and shrimp in urban
area environments are more susceptible to parasitic infections.
4.3.3 Comparisons with Other Studies
4.3.3.1 Fish
Neoplastic and preneoplastic liver lesions in English sole were found
for the first time in Case Inlet. Although the incidence of these lesions
was low (both were 3.3%), previous studies conducted in this embayment
during 1979 and 1980 (a total of 34 fish were examined) did not detect
such lesions. It is not known whether finding these lesions during the
present study reflects a change in the health of English sole in Case
Inlet, or whether a small number of diseased sole were previously present,
but not detected because of the relatively small sample size examined in
past studies (Maiins et al. 1982, 1984).
The absence of detectable serious liver lesions (e.g., neoplasms) in
English sole captured near the mouth of Sinclair Inlet (Stations 17
and 18) is an observation that-agrees with the results of previous
sampling surveys conducted at that location. During eight separate fish
collections performed between 1979 and 1983, no hepatic neoplasms or
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specific degenerative lesions were detected in the 138 English sole that
were examined (Rhodes et al. 1983). A low incidence (1.4%) of hepatic
preneoplasms was found. However, at sampling stations near the upper end
of Sinclair Inlet, in the general area of Station 6, a low incidence of
hepatic neoplasms (0.8%, 1 of 122 fish) and preneoplasms (2.5%, 3 of
122 fish), but no specific degenerative lesions (Maiins et al. 1982) were
found. In the present study, all three types of lesions were found in
English sole from Station 6 at incidences ranging from 6.7% (neoplasms) to
30% (preneoplasms). The results of this study and our previous work
suggest that the incidence of serious hepatic lesions in English sole may
be higher in the upper portion of Sinclair Inlet than the entrance.
Although efforts were made in the field to collect samples of
comparable size, hence age, attaining this goal was not always possible.
Of the English sole collected from the reference sites, 81% were of size
class 1 or 2, which corresponds approximately to the 1+ or 2+ year
classes. This situation was caused primarily by the sole collected at the
Eliza Island site, which displayed little size variation in spite of an
intensive fishing effort. On the other hand, 73% of the English sole
collected in Sinclair Inlet belonged to size class 5, which corresponds
approximately to the 5+ year class. Because the incidence of some
neoplasia has been found to increase with age, care was taken to only
conduct statistical comparisons within individual size (age) classes.
One type of kidney lesion, and depositional disorders, were found
exclusively in Sinclair Inlet. Previous studies in Sinclair Inlet have
identified both these lesions in English sole, but the incidences were not
significantly higher than those at other Puget Sound stations (Maiins
et al. 1982).
Both English sole and Dover sole were examined at the three principal
sampling stations at Fourmile Rock - Elliott Bay dump site vicinity. Even
though only a total of 19 English sole were captured in the bay, a
sufficient number (12) of fish were captured at Station 23 to compare with
previous studies, two sets of sampling stations near Station 23 have
previously been sampled, one station near Magnolia Bluff (NODC No. 10014,
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47° 37' 07" N X 122° 24' 04" W, 15 to 60 m in depth), and a set of
stations referred to as the "Seattle waterfront" stations (Maiins et al.
1984). Station 23 resembled the Magnolia Bluff station in terms of
hepatic neoplasms (none were detected at either station) and preneoplasms
(0% and 1.1%, respectively), but Station 23 was more similar to the
Seattle waterfront in terms of specific degenerative lesions (25.0% and
19.9%, respectively).
Hepatic neoplasms were not detected in Dover sole from Elliott Bay;
however, preneoplastic lesions and specific degenerative lesions of the
liver were found. Preneoplastic lesions were found only in Dover sole
(7.7%, 2 of 30 fish) from Station 17 near the Fourmile Rock - Elliott Bay
dump site, whereas the degenerative lesions were found only in sole (10%,
3 of 30 fish) from the West Point stations. In addition, two lesions of
the gill, necrotic and proliferative conditions, were found only in Dover
sole from this latter station.
Finding important hepatic and gill lesions, which have been shown in
English sole to be pollution-related (Maiins et al. 1984), in Dover sole
from selected stations at the Fourmile Rock - Elliott Bay dump site
vicinity suggests that this species may be adversely affected by
environmental contaminants. However, the low incidence of some of these
lesions, the lack of reference Dover sole, and the lack of information on
the geographical mobility of Dover sole in the sampling area, limit our
ability to further interpret these data.
4.3.3.2 Shellfish
Only two conditions detected in the present study appeared to be
associated with urban bays: hydropic degeneration/membrane lysis in the
hepatopancreas of C. magister and degenerative disorders in the antennal
gland of both shrimp species. These conditions were not associated with
detectable parasitic infections. The incidences of hydropic degenerative/
membrane lysis were highest (33% to 45%) in crab from the inner waterways
of Bellingham Bay, considerably lower (11% to 14%) in Samish Bay, and not
detected at Dungeness Spit.
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The incidence of degenerative disorders in the antennal gland of
shrimp was low, and these disorders were detected only in shrimp from the
Fourmile Rock - Elliott Bay dump site vicinity. These lesions appeared to
be relatively minor, and they would not be expected to seriously impair
function in the affected shrimp.
Pathological conditions have been studied in a variety of Puget Sound
shellfish including crab (Cancer spp.) and shrimp (Crangon alaskensis and
Pandalus spp.) (Maiins et al. 1982). A variety of histopathological
lesions and parasitic infections were identified; however, the
abnormalities were considered to be idiopathic. Necrotic and nodular
lesions had relatively high incidences (in some cases 30% to 80%) in
shrimp and crab collected from urban areas (e.g., Commencement Bay
Waterways, Duwamish Waterway, and the Fourmile Rock - Elliott Bay dump
site vicinity). The incidence of these lesions was generally lower in
organisms collected from nonindustrialized areas.
4.4 AMPHIPOD BIQASSAY
4.4.1 Relationship to Physical and Chemical Properties of Sediments
Correlation analyses illustrated significant relationships between
the amphipod bioassay and various physical and chemical properties of
sediments. Of particular importance were the significant correlations
(p<0.05) of amphipod survival to sediment grain size, percent water,
organic content, and burden of organic compounds.
Amphipod survival was positively correlated with sand content but
negatively correlated with silt and clay content (percent fines) and
organic content (percent volatiles and TOC). Of all the physical and
chemical factors, amphipod survival was the most strongly correlated with
water content of the sediment. Amphipod survival was not correlated with
metals content but was negatively correlated with PCB and AH burdens. Why
sediment burdens of metals (Ag, Hg, Pb) did not correlate with amphipod
survival is not easily explained. Perhaps, the metals existed in
physical-chemical forms that were not generally bioavailable to the
amphipod.
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Although we would like to conclude that lowered amphipod survival
always correlated with high organic loads in the urban bays, we cannot.
It is true that the urban bays demonstrated a higher sediment burden of
organic contaminants, but this did not always result in lowered amphipod
survival (e.g., Fourmile Rock - Elliott Bay dump site vicinity). It is
also true that the baseline bays were found to be relatively clean of most
organics, but this also did not result in higher amphipod survival (e.g.,
Case Inlet, Dabob Bay).
Although direct cause-and-effect relationships between survival and
concentrations of organic compounds cannot be postulated, these data add
credence to the potential for such relationships. Unfortunately, the
state of the art of bioassay development does not allow cause-and-effect
relationships between survival and sediment burden to be rigorously
defined. However, the importance of physical exposure to contaminated
sediments and the potential for ingestion of contaminants associated with
sediments, in particular, appear to be excellent future research
opportunities to elucidate mechanisms of toxic action.
4.4.2 Comparison of Amphipod Survival in Urban and Baseline Bays
Based on amphipod survival data, the urban bays were not always
distinguishable from the baseline bays. While Port Gardner - Everett
Harbor, an urban bay, exhibited the lowest mean survival of 12.3, the
Fourmile Rock - Elliott Bay dump site vicinity, another urban area,
exhibited the highest mean survival of 17.3. Similarly, while Sequim Bay,
a baseline bay, exhibited a mean survival of 17.0, Case Inlet, another
baseline bay, exhibited the very low mean survival of 13.0. Accordingly,
these data would suggest that the baseline or "reference" bay concept is
not as clear-cut as hoped.
The low level of amphipod survival found in Case Inlet deserves
further discussion. Although unexpected, several findings indicate that
the apparent toxicity of Case Inlet sediment is real. First, low mean
survival was detected in two separate surveys (x = 13.6 in the 1983 and
13.0 in 1984). Second, the bioassays were performed by a competent
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laboratory, and no significant "batch" effects were observed. Third, the
control survival for Case Inlet was 19.0, definitely an acceptable level.
Fourth, the physical sediment characteristics in Case Inlet samples were
well within the tolerance limits of the test species, Rhepoxynius
abronius, as determined by Swartz et al. (1984) for percent gravel,
percent sand, percent silt, percent clay, total volatile solids, and
interstitial water salinity. Some low Eh readings were recorded in the
bioassay beakers, but anaerobic conditions probably did not contribute to
the low survival because negative Eh beakers had a higher mean survival
(16.0) than beakers with positive Eh (10.8). Fifth, low amphipod survival
corresponded to low infaunal species richness and abundance in Case Inlet.
However, lowered amphipod survival in Case Inlet may still be the
result of a purely physical response; in this instance, exposure to a
sediment with > 90% silt and clay content. Swartz et al. (1984) presented
particle size limits for individual particle size categories (gravel,
sand, silt, clay). Perhaps the total percentage of fine-grained sediment
(percent silt and clay) should be considered. Applying this convention,
the finest sediment tested by Swartz et al. (1984) was 90% silt-clay.
Many of the Case Inlet stations (4-10) displaying lower survival during
screening surveys had silt-clay percentages ranging from 97 to 99. In
Samish Bay during screening surveys, lowered amphipod survival may also
have resulted from exposure to fine-grained sediments. The three stations
(9, 10, 18) showing the lowest amphipod survival all had silt-clay
contents exceeding 90%. As well, this relationship may explain the
finding of lowered amphipod survival at two stations in Dabob Bay.
It may also be that lowered amphipod survival in both Case Inlet and
Samish Bay resulted from multiple factors: high silt and clay content,
high water content, and slightly elevated metals and organics when
compared to the other baseline bays. Although we have already discussed
the relationship of amphipod survival to percent fines, germane to this
discussion is the observation that 29 of 37 baseline bay stations showing
significantly lowered amphipod survival had sediment water contents
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exceeding 50%. Also, based again on the screening surveys, both Case
Inlet and Samish Bay showed generally higher metals levels and organic
(IR) burdens (Appendix B) when compared with the other baseline bays. The
interpretation is that the level of contamination needed to produce
toxicity in amphipods is somehow less in fine-grained sediments with
higher water contents than that needed to produce the same level of
toxicity in coarse-grained sediments holding less water. Obviously, more
research on how sediment types influence the toxicity of contaminants to
amphipods appears in order and would help address the question of whether
or not sediment quality criteria need to be established for different
sediment types.
4.4.3 Comparisons to Other Studies
The amphipod assay has been applied sparingly in Puget Sound and in
other water bodies. Accordingly, few studies exist which can be
rigorously compared with present findings.
Swartz et al. (1979) conducted sediment bioassays on five different
test organisms including the amphipod, Rhepoxynius abronius, at sites in
the Duwamish Waterway and adjacent sites in Elliott Bay and Puget Sound.
Although f*. abronius experienced significantly higher mortalities at three
of five sediment exposures relative to controls, none of these locations
corresponded to those used in the present studies. Swartz et al. (1982)
also conducted 175 bioassays on Commencement Bay sediments using
R. abronius. They determined that while sediments from central
Commencement Bay, including the two designated disposal sites, were not
acutely toxic, areas of both high and low toxicity were detected in the
waterways and other nearshore areas of Commencement Bay. Chapman et al.
(1982), under NOAA sponsorship, conducted a comprehensive sediment
bioassay program at 97 sites in Elliott Bay, Commencement Bay, Sinclair
Inlet, Port Madison, and Birch Bay. While R. abronius among other test
organisms was used, the test procedures called for exposures to water and
sediment slurries. In contrast to Swartz et al. (1982), and to the
present studies, Chapman et al. (1982) showed essentially no acutely
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lethal conditions at the sites surveyed. The only exception was when
sediments from a site near the Denny Way (Elliott Bay) combined
sewer outflow were assayed. Ott1 also, conducted amphipod bioassays of
Puget Sound sediments. Both R. abronius and Eohaustorius washingtonianus
were exposed directly for 240 h to 17 test sediments in a recirculating
seawater system. Although there also occurred a spatial heterogeneity in
the toxic response within localized areas, sediments from four sites (one
from Elliott Bay, two from the Duwamish Waterway and one from Sinclair
Inlet) resulted in significantly lower survival than the control sediment
stations. More recently, Chapman et al. (1984b) surveyed sediments from
22 stations in Everett Harbor, Bellingham Bay, and Samish Bay using the
amphipod bioassay. Two stations (one each in Bellingham Bay and Everett
Harbor) demonstrated significant acute lethal effects, while the two
Samish Bay reference stations showed no significant effects.
It should also be noted that in the present study, all sediments were
assayed fresh (unfrozen). Some of the previous studies (Chapman et al.,
1982, 1984a) were conducted with frozen sediments which may have altered
both bioavailability and toxicity. Contaminants may have been partitioned
and toxicity seriously underestimated. .This problem has been discussed
recently by Lee and Jones (1982) in relation to dredged material
evaluations. This and other evidence (Stober et al. 1983) suggest that
there is no substitute for performing either chemical or biological
analyses on fresh sediment samples.
4.5 OYSTER LARVAL BIOASSAY
4.5.1 Relationship to Physical and Chemical Properties of Sediments
Although we would like to state that a higher percentage of abnormal
larvae always correlated with high metals IR, TOC, or specific organics in
the urban bays, we cannot. The urban bays did demonstrate a higher
sediment burden of metals and organic chemicals, but this burden did not
always result in a higher percentage of abnormal larvae. The baseline •
1 Personal communication, 1982. Unpublished information from a study with
P.O. Plesha, R.D. Bates, C. Smith, and B.B. McCain, "An Evaluation of an
Amphipod Sediment Bioassay Using Sediment from Puget Sound."
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bays were found to be relatively clean of metals and chemicals, but did
not always have a lower percentage of abnormal larvae. Obviously, both
fine-grained and high-water-content sediments are somehow involved.
However, we cannot deduce that fine-grained and high-water-content
sediments per se produce increased percentages of abnormal larvae.
Rather, several factors likely account for the degree of variance
measured, and grain size and percent water are but two of the factors.
This situation does not rule out the involvement of sediment chemistry in
some way. However, because bays were sometimes tested in different assays
and at different times, more rigorous comparisons are confounded by batch
and time effects.
4.5.2 Comparison of Oyster Larval Abnormalities in Urban and Baseline
Bays
Based on percent abnormality in oyster larvae, the urban bays were
not always distinct from the baseline bays. Although a majority of
stations in Bellingham Bay and Sinclair Inlet exhibited significantly
higher mean percentages of abnormal larvae when compared with controls,
the Fourmile Rock - Elliott Bay dump site vicinity, another urbanized
area, exhibited no such differences. Similarly, although Sequim Bay,
Samish Bay, and Case Inlet, all baseline bays, exhibited no differences
among mean percentages of abnormal larvae when compared to controls, Dabob
Bay, another baseline bay, did exhibit a higher mean percentage of
abnormal oyster larvae when compared to controls. Accordingly, these
data also suggest that the baseline or "reference" bay concept may not
always be as helpful as first believed.
4.5.3 Comparisons to Other Studies
The oyster larval bioassay has not been routinely applied to the
assessment of sediment toxicity. Accordingly, as in the case of the
amphipod bioassay, relatively few studies exist that can be compared with
the present findings.
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Chapman and Morgan (1983) tested sediments collected from 22 stations
in Puget Sound, including five stations in Elliott Bay and two stations in
Sinclair Inlet. Although the station locations at Fourmile Rock - Elliott
Bay and Sinclair Inlet in the two surveys did not match exactly, they
were, nevertheless, in the same general vicinity. The two surveys were in
general agreement that Sinclair Inlet stations are more toxic than
Fourmile Rock - Elliott Bay stations. More recently, Chapman et al.
(1984b) conducted oyster larval bioassays on sediments from 22 stations in
Everett Harbor, Bellingham Bay, and Samish Bay. A total of 19 stations in
the two urban-industrialized bays demonstrated significant abnormalities
and mortalities, while the two reference stations in Samish Bay showed no
significant effects. Again while station locations in the two surveys did
not exactly match, they were, however, in the same general vicinity.
Accordingly, we would expect that the two surveys would be in general
agreement that both Bellingham Bay and Everett Harbor stations were more
toxic than the reference stations in Samish Bay. The fact that we failed
to detect toxicity at Port Gardner - Everett Harbor, particularly in the
East Waterway, is not easily explained. One possible explanation is that
relatively high variability and poor performance in control cultures in
Assay No. 3 did not allow for detection of statistical differences among
treatments.
4.6 SUMMARY STATISTICS
The scope of the present study did not provide for recommendations as
to which remedial actions were required in the most impacted bays, but it
did provide for the identification of those bays that now show signs of
degraded sediment quality (Table 60). This exercise could help determine
which bay to concentrate further research, mitigation, and/or clean-up
efforts. These data, when taken in concert, demonstrated that the urban
bays were significantly more impacted than the baseline bays. These data
also indicated that the greatest impacts were found in Port Gardner -
Everett Harbor, Sinclair Inlet, and Bellingham Bay. The Fourmile Rock -
Elliot Bay dump site vicinity appeared to be the least impacted of the
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urban-industrialized sampling areas. As well, and not surprisingly,
Sequim Bay was the least impacted of baseline bays. However, it was
easily seen from these data that Case Inlet and Samish Bay showed some
indication of degrading sediment quality. All stations in each bay
resulted in lower than mean control amphipod survival. The data for Dabob
Bay also suggest some degree of impact.
Inspection of the data in Table 60 shows that the most adversely
impacted stations in Port Gardner - Everett Harbor were located in the
East Waterway. For Sinclair Inlet, they were close to the Puget Sound
Naval Shipyard. In Bellingham Bay, they were associated with the Whatcom
Creek and I and J Street waterways.
Although the fish and shellfish pathology data were not used in this
summary statistical exercise, (see Section 2.8.5) they would serve to
corroborate our results: English sole caught in Sinclair Inlet
demonstrated the highest incidences of liver, kidney, and gill lesions.
Also, two types of lesions were found only in animals from urban areas.
Hydropic degeneration/membrane lysis in the hepatopancreas of Dungeness
crab and degenerative disorders in the antennal gland of both shrimp
species were observed in Bellingham Bay and in the Fourmile Rock - Elliott
Bay dump site vicinity.
Finding a biological signal (significantly lowered amphipod survival)
in Case Inlet and Samish Bay is not easily explained. The results of
sediment chemistry analyses are not as clear-cut as those for Sinclair
Inlet and Port Gardner - Everett Harbor or Bellingham Bay. The slightly
elevated silver concentrations in both Screening and Detailed Surveys in
Case Inlet could indicate the presence of sewage contamination,
Particulate contaminants could enter Case Inlet through transport
processes from the central basin of Puget Sound, or runoff from septic
tank drain fields, and animal pastures. Certain base-neutral organic
compounds found in Samish Bay during the detailed surveys perhaps suggest
transport of industrial wastes from Bellingham Bay. More disturbing is
that specific locations sampled in Case Inlet and Samish Bay were selected
as generally the cleaner of the stations. Accordingly, the problem in
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these embayments could be much worse. Whatever the explanation, much
additional research must be conducted.
4.7 RELATIONSHIP OF URBAN TO BASELINE (REFERENCE) BAY CONCEPT
It would seem easy to describe baseline or "reference bay" conditions
based on chemical variables. In this study, clear differences occurred in
urban versus baseline bays as to concentrations of metals, PCB-1254, and
aromatic hydrocarbons. Sequim Bay and Dabob Bay, with relatively low
metals and no detectable organic compounds, appear to be excellent choices
for reference bays.
However, because all marine waters of Puget Sound and the Strait of
Juan de Fuca interconnect, contamination from one bay will result in some
degree of contamination to all bays. This fact and the finding of low
levels of chemical contaminants in some of the baseline bays (Samish Bay,
Case Inlet) in this study would suggest that the concept of "reference
bay" is not as clear-cut as first believed, and that perhaps what we have
observed is evidence of a continuum or gradient of contaminants extending
from the urban bays.
On the basis of biological variables, the concept of baseline or
"reference bay" is also less clear, and perhaps is better described as
representing a continuum of responses consistent with a gradient of
contaminants existing between regions of higher and lower contamination in
Puget Sound. The results of both the amphipod and oyster larval bioassays
tend to best support this interpretation. Although Port Gardner-Everett
Harbor, Bellingham Bay, and Sinclair Inlet, all urban bays, exhibited
generally lowered amphipod survival and/or higher oyster larval
abnormality, the Fourmile Rock-Elliott Bay dump site vicinity, another
urban area, exhibited the highest mean amphipod survival and no difference
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in percentage of oyster larval abnormality when compared with controls.
Similarly, Case Inlet, Samish Bay, and Dabob Bay, all baseline bays,
showed either a relatively low amphipod survival or a higher than control
percentage of abnormal oyster larvae at no less than half of the stations
tested. The data could be interpreted as meaning: 1. Port Gardner -
Everett Harbor, Bellingham Bay and Sinclair Inlet are all typically urban
bays that in Puget Sound are sources of chemical contamination. They
occupy one end of the continuum. 2. Sequim Bay is typically a baseline
bay located at the opposite end of the continuum far from urban sources of
contamination, 3. Fourmile Rock - Elliott Bay is intermediate on the
continuum located relatively close to an urban source of contamination -
Elliott Bay. However, clearly the dump site is unique because not one of
the eight stations in the present study indicated degraded sediment
quality when evaluated by either the amphipod or oyster larval bioassay.
4. Case Inlet and Samish Bay are also located intermediate on the
continuum but relatively far from urban sources of contamination and
closer to Sequim Bay conditions. However, both are close enough to
Commencement Bay and Bellingham Bay respectively to still show some
degraded sediments. 5) Dabob Bay is also located intermediate on the
continuum but closest to the Sequim Bay positon, that is, relatively far
from most urban sources of contamination. The finding of some suggestion
of degraded sediment quality in Dabob Bay is perhaps attributed to
circulation of contaminants from the main basin of Puget Sound.
The infaunal data also suggest that the concept of baseline or
reference bay is less clear-cut. In particular, the differences in
physical characteristics (depth, grain size) among urban and baseline bays
greatly influenced indigenous infaunal communities (species richness and
abundance), and necessarily led to comparisons within sediment type.
Actually, the bays differed in their depth, sediment type, etc., with as
much variation within the urban and baseline categories as among all bays.
It was also evident that the influence of sediment chemistry on
infauna was difficult to discern. Against the variation in infaunal
indices attributed to physical factors, the influence of only one chemical
variable, TOC, was consistently observed. In all sediment types, high TOC
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4.0 to 16% was associated with a disturbed infauna dominated by organic
enrichment opportunist species. This shift to enrichment opportunists
occurred at almost all stations in Port Gardner - Everett Harbor and at
several stations in Bellingham Bay. The extent of the shift was less in
sandy sediments when compared with silty and muddy sediments. In
addition, stations with the most impoverished infauna occurred both in
urban and baseline bays and were associated with greater depth and higher
clay content rather than with higher concentrations of contaminants.
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5.0 CONCLUSIONS
5.1 BATHYMETRY AND SEDIMENT CHARACTERISTICS
The eight bays demonstrated different depth and sediment
characteristics. The Fourmile Rock - Elliott Bay dump site vicinity and
Dabob Bay were distinctly deeper (>100 m) and contained sediments which
were, on the average, sandier and had lower total organic carbon (TOC)
contents than all the rest. Samish and Bellingham Bays were the
shallowest and the muddiest of the eight bays. Bellingham Bay showed the
second highest TOC level. Case Inlet, Sinclair Inlet, and Sequim Bay were
of intermediate depth, and had intermediate values for percentage of silt,
clays, and TOC. These three bays exhibited sediment types ranging from
sandy-silt to silty-mud. Port Gardner - Everett Harbor was distinct from
the other bays in its significantly higher mean TOC level.
5.2 CONTAMINANT LEVELS
Urban bay sediments, particularly in Sinclair Inlet and at the
Fourmile Rock - Elliott Bay dump site vicinity, were found to be
substantially contaminated by Ag, As, Cu, Cd, Hg, Pb, and Zn. Other
metals (Sb, Be, Cr, Ni, Se, and Ti) were found at about the same
concentration in all bays; however, a few urban sediments in Bellingham
Bay and Sinclair Inlet were enriched in Sb, Cr, and Ni. Baseline bays
generally contained lower concentrations of metals, many of which were
found at approximately the same concentrations as in pre-1900 sediments.
Urban bay sediments, particularly Sinclair Inlet and Port Gardner -
Everett Harbor, also contained significant quantities of aromatic
hydrocarbons and PCB-1254. Other organic compounds occasionally found in
urban bays included phthalates and PCB-1260. However, no PCBs were
detected in baseline bays, and only a few stations in Samish Bay contained
detectable quantities of phthalates and aromatic hydrocarbons.
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5.3 BENTHIC INFAUNA
It was determined that in all sediment types, TOC levels between 4
and 16% were correlated with a disturbed infauna dominated by organic
enrichment opportunists. This shift to enrichment opportunists, primarily
nematodes and Capitella capitata, occurred at almost all stations in Port
Gardner - Everett Harbor and at several stations in Bellingham Bay. The
extent of this shift was less in sandy sediments when compared to silty or
mud sediments.
After omitting stations with high values of TOC, the numerically
dominant infauna were shown to vary more with sediment type than by bay,
except that Case Inlet and Samish Bay demonstrated distinctly different
infaunas from all other bays. Also, omitting these same stations did not
reveal a distinctly different set of numerically dominant species in urban
bays when compared with baseline bays.
5.4 FISH AND SHELLFISH PATHOLOGY
Because flatfish species were not collected in four of the eight bays
(Bellingham Bay, Samish Bay, Dabob Bay, Sequim Bay) because of seasonal
scarcity, the resulting pathology data were not used in the summary
statistical exercise (Section 2.8.5) to determine signs of degraded
sediment quality. However, these data did serve to corroborate the
results of the approaches used in the summary exercise (chemistry,
infaunal analyses, amphipod and oyster larval bioassay), indicating that
urban bays were generally more impacted than baseline bays.
English sole caught in Sinclair Inlet demonstrated the highest
incidence of liver, kidney, and gill lesions. Also, two types of lesions
in shellfish were only found in animals from urban bays. Hydropic
degeneration/membrane lysis in the hepatopancreas of Dungeness crab and
degenerative disorders in the antennal gland of two shrimp species were
observed in Bellingham Bay and at the Fourmile Rock - Elliott Bay dump
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site vicinity. And, although the incidence of serious liver, kidney, and
gill lesions in English sole from.Case Inlet were low compared to the
incidence detected in sole from other urban bays (Duwamish Waterway),
these lesions were not detected in sole from Eliza Island.
5.5 AMPHIPOD SURVIVAL
Based on the amphipod survival data alone, the urban bays were not
always distinguishable from baseline bays. The Fourmile Rock - Elliott
Bay dump site vicinity, selected as representative of urban bay
conditions, and Sequim Bay, a baseline bay, demonstrated the highest and
second highest mean survivals of 17.3 and 17.0 respectively. Similarly,
Case Inlet, a baseline bay, and Port Gardner - Everett Harbor, an urban
bay, demonstrated the second lowest and lowest mean survivals of only 13.0
and 12.3, respectively.
Correlation analyses illustrated significant relationships between
the amphipod bioassay and various physical and chemical properties of
sediments. Of particular importance were how amphipod survival related to
sediment grain size, percent water, and burden of organic compounds
(PCB-1254, aromatic hydrocarbons, and IR). All such properties were
interpreted as accounting for lower amphipod survival. These analyses
also revealed that lowered amphipod survival occurred in sediments of Port
Gardner - Everett Harbor where an increased abundance of organic
enrichment infaunal species was encountered.
5.6 OYSTER LARVAL BIOASSAY
Results of the oyster larval bioassay generally paralleled those of
amphipod bioassays. The analyses, when taken alone, indicated that the
urban bays were not always distinguished from those of baseline bays.
Although a majority of stations in Bellingham Bay and Sinclair Inlet, both
urban bays, exhibited significantly higher mean percentages of abnormal
larvae when compared with controls, the Fourmile Rock - Elliott Bay dump
site vicinity (also considered representative of urban bay conditions)
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exhibited no differences when compared with controls. Similarly, although
Sequim Bay, Samish Bay, and Case Inlet, all baseline bays, exhibited no
differences among mean percentages of abnormal larvae when compared with
controls, Dabob Bay, another baseline bay, exhibited higher mean
percentages of abnormal larvae at two of its four stations.
Correlation analyses showed a significant relationship between
percent abnormal oyster larvae and amphipod survival. The trend to higher
percentages of abnormal oyster larvae becomes more evident as amphipod
survival decreased.
5.7 SUMMARY STATISTICS
On the basis of the summary exercise which evaluated all data
(chemical and biological), urban bays were significantly more impacted
than baseline bays. The greatest impacts were found in Port Gardner -
Everett Harbor, Sinclair Inlet, and Bellingham Bay. The Fourmile Rock -
Elliott Bay dump site vicinity appeared to be the least impacted of the
urban-industrialized sampling areas. Not surprisingly, Sequim Bay was the
least impacted of the baseline bays. However, Case Inlet and Samish Bay,
both baseline bays, showed some indication of degraded sediment quality.
All sediments in both bays resulted in lower than control amphipod
survival. The amphipod and oyster larvae bioassay data from Dabob Bay
also suggested some degree of impact.
It was also observed that the most adversely impacted stations in
Port Gardner - Everett Harbor were located in the East Waterway. For
Sinclair Inlet, they were located near the Puget Sound Naval Shipyard. In
Bellingham Bay, they were associated with the inner harbor. Case Inlet
demonstrated the second lowest species richness, the lowest infaunal
abundance, and the lowest amphipod survival of all eight bays. Samish Bay
was found to contain phthalates and aromatic hydrocarbons and demonstrated
the third lowest amphipod survival of all eight bays.
Clearly, the strength of this exercise was in the performance of
several analyses, both chemical and biological, on the same sediment
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sample. The development of a broad based (summary) index of degraded
sediment quality also facilitated detection of a wider range of sediment
toxicity that would not have been possible if a single analysis or
bioassay had been applied.
5.8 RELATIONSHIP OF URBAN TO BASELINE (REFERENCE) BAY CONCEPT
On the basis of chemical contaminants alone, the baseline or
"reference bay" concept appears to be valid. Sequim Bay and Dabob Bay,
with relatively low metals burdens and no detectable organic compounds,
seem to be excellent choices for reference bays. Application of this
concept is potentially useful in Puget Sound in a regulatory sense to
determine where and when remedial action is required.
Biological variables, however, do not always clearly distinguish
between what we hypothesized at the outset of the study to be urban bays
and baseline bays. Rather, the biological findings better describe a
continuum of responses consistent with a gradient of contaminants existing
between regions of higher and lower contamination in Puget Sound.
This continuum is perhaps best described as follows: 1) Port Gardner
- Everett Harbor, Bellingham Bay, and Sinclair Inlet occupy one end of the
continuum representing water bodies now showing the most evidence of
degraded sediment quality. 2) Sequim Bay is a baseline bay located at the
opposite end of the continuum. It currently shows the least indication of
degraded sediment quality of all the bays sampled. 3) The Fourmile Rock -
Elliott Bay dump site vicinity is located intermediate on the continuum,
but closer to the position occupied by the urban bays. However, the dump
site is somewhat anomalous because not one of the eight stations sampled
during the detailed surveys showed any signs of degraded sediment quality
when evaluated by either the amphipod or oyster larval bioassay. 4) Case
Inlet and Samish Bay are also located intermediate on the continuum but
closer to the baseline bay position occupied by Sequim Bay.
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Both bays, however, are close enough to Commencement Bay and Bellingham
Bay, respectively, to show some signs of degraded sediments. 5) Dabob Bay
is also located intermediate on the continuum, but closest to the baseline
bay position occupied by Sequim Bay. The finding of degraded sediment
quality based again on the bioassay results is perhaps attributed to
circulation of contaminants from the main basin of Puget Sound.
In summary, the results of our study support the "reference bay"
concept; however, our results also suggest that the process to select
reference bays needs refinement. It is evident that selection should be
based on chemical, biological, and physical data. Also, the 1983
screening survey was particulary useful in providing relevant and recent
data on which to base the selection process. Ideally, a reference bay
should contain little or no chemical contamination, demonstrate few or no
impacts when evaluated by infaunal analyses and sediment bioassays, but
should also possess physical characteristics (depth and grain size) that
closely approximate the urban bay(s) to be examined. Our results perhaps
suggest that "reference stations" may be a more appropriate concept than
"reference bays", particularly when time and funds limit the scope of
studies performed in support of regulatory action.
There is a need to bring together a forum of scientists and
regulators to refine this concept and its intended use. Also, our strategy
of selecting equal numbers of the most contaminated stations within each
urban bay does not address issues concerning the full range of
contaminated sediments within each bay. Clearly, the present study would
have benefitted from the addition of stations both in the urban and
baseline bays. Consequently, our results apply only to the sediments
(stations) sampled in each bay and do not likely represent all similiar
depositional sediments in each bay.
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5.9 USEFULNESS OF THE RECONNAISSANCE SURVEY CONCEPT
The 1983 screening survey of 181 stations from eight bays was
particularly useful in appropriately focusing the 1984 detailed survey.
The strategy of selecting the most contaminated stations from the urban
bays and the least contaminated stations from the baseline bays permitted
limited resources to be directed toward identifying those stations most
likely to reveal degraded sediment quality. Moreover, the screening
survey data allowed the station selection process for the detailed surveys
to be based on data that was recently acquired rather than on historical
data that was collected in support of other goals. The variability
encountered in biological indices subsequently revealed the wisdom of the
selection strategy applied. Another selection strategy would have likely
provided less clear-cut biological findings. Consequently, we have
learned that sampling strategies must be developed specifically for each
study; and if faced with another large sediment survey, we would again
recommend that a preliminary or reconnaissance level effort be undertaken
to determine which sampling strategy best addressed the study goals within
available resources.
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