Puget Sound Estuary Program
SAMPLING AND ANALYSIS DESIGN
FOR DEVELOPMENT OF
EVERETT HARBOR ACTION PROGRAM
DRAFT REPORT
PREPARED BY:
TETRA TECH, INC.
FOR:
U.S. ENVIRONMENTAL PROTECTION AGENCY
REGION X - OFFICE OF PUGET SOUND
SEATTLE, WASHINGTON
JULY, 1986
-------
Draft Report
TC 3991-03
SAMPLING AND ANALYSIS DESIGN
FOR DEVELOPMENT OF EVERETT HARBOR
ACTION PROGRAM
by
Tetra Tech, Inc.
for
U.S. Environmental Protection Agency
Region X - Office of Puget Sound
Seattle, Washington
July, 1986
Tetra Tech, Inc.
11820 Northup Way, Suite 100
Bellevue, Washington 98005
-------
CONTENTS
Page
LIST OF FIGURES iii
LIST OF TABLES iv
INTRODUCTION 1
GENERAL APPROACH 3
STUDY TYPES AND INTEGRATION 3
SPATIAL ANALYSIS 4
REFERENCE AREA 6
CRUISE PROCEDURES 7
STATION LOCATION METHODS 7
SEDIMENT QUALITY SURVEY 9
DATA GAPS 9
GENERAL STUDY DESIGN 11
STATION LOCATIONS 13
SAMPLING METHODS, PROCESSING, AND ANALYSES 17
SEDIMENT BIOASSAYS AND BENTHIC MACROINVERTEBRATE COMMUNITIES 20
DATA GAPS 20
GENERAL STUDY DESIGN 21
STATION LOCATIONS 25
SAMPLING METHODS AND SAMPLE PROCESSING 26
LABORATORY PROCEDURES 27
BIOACCUMULATION AND PATHOLOGY 29
DATA GAPS 30
GENERAL STUDY DESIGN 31
STATION LOCATIONS 36
SAMPLING METHODS 37
SAMPLE PROCESSING 38
DATA MANAGEMENT 40
SUMMARY 41
REFERENCES 42
APPENDIX: MAPS
ii
-------
FIGURES
NOTE: Figure found immediately following page indicated
Number Page
1 Project area: Everett Harbor and the Lower Snohomish River 1
2 General approach to development of Everett Harbor Action Plan 1
3 Components of recommended study design 3
4 Sample processing scheme for pathology and bioaccumulation
component 38
-------
TABLES
NOTE: Table found immediately following page indicated
Number Page
1 Determination of minimum detection levels for elevated
incidence of disease given 10 sample sizes and three back-
ground levels of disease 34
2 Summary of basic study design 41
-------
INTRODUCTION
Under the Puget Sound Estuary Program, the U.S. Environmental Protection
Agency (EPA) and the Washington Department of Ecology in cooperation with
other local, state, and federal agencies are developing an action plan
to solve problems of toxic and microbial contamination in Port Gardner
and the lower Snohomish River. The resulting Everett Harbor Action Plan
will: 1) identify existing problems of toxic chemical contamination, microbial
contamination, and associated biological effects within the study area
boundaries (Figure 1); 2) identify ongoing and historical sources of toxic
chemical pollution; and 3) identify appropriate remedial actions and agency
responsibilities for implementing the foregoing actions. Recommendations
for short-term corrective actions will be presented in the Interim Work
Plan. The sampling and analysis plan presented here represents the study
design for a field investigation to supplement the existing database.
Results of the field studies will be used to develop the final action plan
(Figure 2). Recent historical data and results from the studies described
herein will be integrated into a decision-making process to identify problem
areas and rank them in terms of priority for action. Thus, study areas
defined in Figure 1 will be used in the data evaluation stage, but all
areas will not be sampled as part of this project. Elements of the decision-
making approach and summaries of available data are provided in the Initial
Data Summaries and Problem Identification report (Tetra Tech 1985b). The
conceptual basis of the Everett Harbor Action Program and the sampling
and analysis specifications described herein are similar to those used
previously for the Commencement Bay Superfund Project (Tetra Tech 1985a)
and the Elliott Bay Toxics Action Plan (Tetra Tech 1985c).
Each major section of this sampling and analysis plan describes a
study design component, including study objectives, kinds of samples, variables
to be measured, sampling methods, sample processing, and laboratory analyses.
The Quality Assurance Project Plan (Tetra Tech 1986b) provides details
-------
EVERETT HARBOR AREAS
MUKILTEO
^^^^^••^^ NAUTICAL MILES
KILOMETERS
2 CONTOURS IN FEET
EAST WATERWAY
SOUTH PORT GARDNER
(T) OFFSHORE PORT GARDNER
(7) SNOHOMISH RIVER DELTA
(?) SNOHOMISH RIVER
(£) PORT GARDNER DISPOSAL SITE
@ EBEY SLOUGH
(?) STEAMBOAT SLOUGH
(9) UNION SLOUGH
Figure 1. Everett Harbor project area.
-------
DATA COLLECTION
DATA EVALUATION
PROBLEM-AREA
EVALUATION
POLLUTION SOURCE
EVALUATION
REMEDIAL ACTION
PLAN
DATA
GAPS
FIELD STUDY
DESIGN
I
Figure 2. General approach to development of Everett Harbor
Action Plan.
-------
of sample handling, analytical chemistry, and other Quality Assurance/Quality
Control (QA/QC) procedures.
-------
GENERAL APPROACH
The general objectives of the study design are to:
0 Identify problem areas related to chemical contamination
• Define the spatial extent and chemical/biological characteristics
of problem areas
• Prioritize problem areas for remedial action.
The approach used to develop the study design involved: 1) evaluation
of existing data for each study component (e.g., sediment chemistry, benthic
infauna); 2) mapping of station locations with acceptable historical data;
3) preliminary assessment of problem areas based on spatial and temporal
distribution of pollutant sources, contamination of sediments, and biological
effects; 4) identification of data gaps in terms of spatial and temporal
coverage of existing data; and 5) selection of study components, conceptual
approach, specific variables to be measured, and station locations for
this study plan. Sampling stations for each recommended study component
were positioned to fill gaps in the spatial coverage of previous studies,
to ensure adequate characterization of areas near major pollutant sources,
and to confirm selected problem areas identified in previous studies.
All maps referred to in the text are in Appendix A.
STUDY TYPES AND INTEGRATION
Components of the study design are shown in Figure 3. Each study
component will provide data for a specific environmental indicator (e.g.,
sediment chemistry, benthic infauna) relevant to the problem identification
process. The rationale for the choice of indicators, their interrelationships,
-------
SURVEY OF CONTAMINATION AND
BIOLOGICAL EFFECTS IN
EVERETT HARBOR STUDY AREA
SEDIMENT QUALITY
• Surface Sediment
Chemistry
• Grain-Size
• Subtidal and Intertidal
SEDIMENT BIOASSAYS
• Amphipod Survival
• Subtidal and Intertidal
BENTHIC INFAUNA
• Community Structure
• Subtidal
BIG-ACCUMULATION
• Fish Muscle
• Crab Muscle
• Subtidal
FISH PATHOLOGY
• External
• Liver
• Subtidal
Figure 3. Components of recommended study design.
-------
and action-level criteria that define problem areas are described in the
Initial Data Summaries and Problem Identification report (Tetra Tech 1985b).
Concurrent sampling of related variables can help ensure timely completion
of the project, efficient to use of cruise resources, and collection of
appropriate data. The general timing of the field work is outlined below:
• August — Bioaccumulation
— Pathology
• September/October — Sediment Quality
— Benthic Infauna
— Bioassays
Timing of individual surveys is justified below in the discussion of detailed
study designs. Briefly, the comprehensive benthic survey is scheduled
after the major recruitment period (spring-summer) for many infaunal species.
The timing of the bioaccumulation and pathology study ensures that English
sole will have resided in shallow-water sampling locations for several
months before sampling.
SPATIAL ANALYSIS
Discrimination of spatial patterns in contaminant distributions and
biological responses is a major objective of this project. To facilitate
spatial analysis, the project area has been divided into nine smaller areas
based on geographic features, bathymetry, and locations of major pollutant
sources (Figure 1). The East Waterway was defined as a distinct area.
The Snohomish River and Estuary includes five areas. The remaining portion
of Port Gardner includes three areas: the deep, offshore areas; the southern
shoreline; and the Port Gardner Disposal Site. Area boundaries and major
features are as follows:
-------
1. East Waterway—All of the East Waterway north and east of
a line from the Snohomish River mouth black can buoy "3A"
to the southern-most boundary of the former Weyerhaeuser
pulp mill dock.
2. South Port Gardnei—Shoreline areas (<30 ft deep) from Elliot
Point (Mukilteo) to the southernmost boundary of the former
Weyerhaeuser pulp mill dock
3. Offshore Port Gardner—All deep-water (>30 ft) areas of
Port Gardner exclusive of the Port Gardner Disposal Site
(see No. 6 below).
4. Snohomish River Delta—The area west of a line between the
downstream boundary of Ebey and Smith Islands out to the
30-ft depth contour.
5. Snohomish River—The main navigable river channel downstream
from the Interstate-5 (1-5) bridge to the mouth of the river
(marked by the black can bouy "3A").
6. Port Gardner Disposal Site—The designated disposal site
in South Port Gardner.
7. Ebey Slough—The channel adjacent to the northern boundary
of Ebey Island, west of 1-5 to a line downstream between
Priest Point and the western tip of Ebey Island.
8. Steamboat Slough—The channel between Ebey and Smith Islands,
west of 1-5 to a line between the western tip of Ebey Island
and the northwestern tip of Smith Island.
<
9. Union Slough—The portion of the slough west and north of
1-5.
-------
During the data analysis phase, some of the above areas may be subdivided
based on observed distributions of contamination and effects.
Several approaches to spatial analysis will be used: 1) assessment
of contamination/response at individual stations for detection of "hot
spots" (i.e., relatively localized areas of contamination and biological
effects); 2) gradient analysis; 3) comparisons among areas for input to
the priority ranking procedure; and 4) comparisons of individual stations
and area averages with data from reference sites. Each approach will be
important for assessing the distribution of contamination and biological
effects in the project area. If major sites .of contamination and effects
are found beyond the influence of all known sources, other causes will
be investigated (e.g., historical contamination, unidentified local source,
undefined transport process).
REFERENCE AREA
The ideal reference area exhibits physical characteristics (e.g.,
sediment type, water depth, wave exposure, freshwater influence) that are
similar to those of the study area, but is without significant chemical
contamination. The upper portion of Port Susan will be used as a reference
area for comparison with the Everett Harbor study area. The reference
area is located approximately 20 km northwest of the study area. The physical
characteristics of Port Susan appear to be similar to those of inner Everett
Harbor. Port Susan has muddy to sandy sediments at shallow depths, is
a partially enclosed embayment, and is influenced by freshwater from a
major river, the Stillaguamish. Data on the biota (i.e., fishes, benthos)
of upper Port Susan are not available at present. Limited data on sediment
chemistry indicate that contaminant levels for total aromatic hydrocarbons,
polychlorinated biphenyls (PCBs), chlorinated butadienes, hexachlorobutadienes,
and most metals are low (Mai ins et al. 1982). Sediment concentrations
of mercury are elevated slightly, however. Additional data on sediment
chemistry (e.g., pesticides, acids, bases, volatile solids) are not available.
-------
CRUISE PROCEDURES
Standard methods of operation will be developed as part of the cruise
plan. They incorporate QA/QC procedures consistent with the Quality Assurance
Plan for this project. Special precautions will be taken to prevent contami-
nation of samples during collection and initial processing aboard the survey
vessel. Cleaning of samples, working areas, and instruments before collection
of each sample for chemical analysis is essential. Work areas of the ship
will be arranged to avoid contamination of samples by engine exhaust, oil,
and other interfering substances. Details of QA/QC procedures are provided
in the Quality Assurance Project Plan (Tetra Tech 1986b).
STATION LOCATION METHODS
Because of the many sources and documented spatial heterogeneity of
contamination in the Everett Harbor study area, precise positioning of
sampling stations is essential. The intent of the navigational control
effort is to accurately determine and document the locations of all sampling
stations and transects. In the Snohomish River and inner portion of Everett
Harbor this is complicated because routine electronic navigation equipment
(e.g., microwave units or Loran C) will not function accurately. At the
same time, horizontal distances to fixed shore objects are short, and there
are many fixed points available for referencing station locations. During
the cruises, the available visual reference points in the Snohomish River
and the East Waterway will be photographically recorded (i.e., corners
of buildings and piers, spires, towers, smoke stacks, and other readily
distinguishable, permanent objects). The shipboard photos will be compared
to aerial photographs and USGS quadrangle maps for the area. Objects that
can be recognized clearly on the aerial photograph or map, and hence can
be accurately located, will be selected and numbered as reference points.
The series of shipboard photos, with the reference points identified and
numbered, will constitute the primary station-location tool in the waterways.
In practice, stations will be located by establishing ranges between
two reference points, if possible, and/or by establishing along-channel
-------
and cross-channel distances to shore objects. All station locations will
be documented by photographs taken at the time samples are collected and
by written descriptions of relationships to the reference points. Station
positioning methods will be accurate enough to define locations within
approximately a 10-ft radius. Standardized procedures will be used throughout
the project. In the outer portions of the study area, Loran C wi11 be
used as the primary positioning tool, with line-of-site and photographic
confirmation.
The boat will be anchored at stations whenever possible, and station
locations will be verified just before each sample is taken. The plotted
station locations will be converted to state plane coordinates for entry
into the database.
8
-------
SEDIMENT QUALITY SURVEY
The detailed study design for the sediment quality survey is presented
in this section, following a brief summary of data gaps.
DATA GAPS
Most of the available information on toxic chemical contamination
of the Everett Harbor study area is based on analyses of sediment quality.
Station locations for the accepted data sets obtained from previous studies
are shown in Maps 3 and 4. Only in a few areas are the data sufficient
to indicate whether problems may exist, and even in those areas the sampling
has not been oriented toward developing relationships between high levels
of toxic chemicals and sources. Compared with other areas of Puget Sound,
the total suite of chemicals measured in previous studies in Everett Harbor
has been limited [i.e. PCBs, polynuclear aromatic hydrocarbons (PAH), and
metals only]. Specific data gaps for each of the areas are discussed below.
East Waterway
East Waterway has been sampled extensively (Map 4). A large number
of grab samples and deep cores have adequately characterized the horizontal
and vertical distributions of the selected indicator chemicals used in
the analysis below (i.e., PAH, PCBs, and selected metals). However, information
regarding the concentrations and distributions of organic compounds other
than PAH and PCBs is needed. Of particular interest are those substances
that may be associated specifically with the pulp mills, such as the phenolic
and chlorinated phenolic compounds. Results of recent studies in Commencement
Bay (Tetra Tech 1985a) and other studies suggest that the following substances
of concern may accumulate in sediments as a result of pulp-mill discharges:
-------
• Monochloro- and dichlorodehydroabietic acid
t Dehydroabietic acid
• Isopimaric acid
• 3,4,5- and 4,5,6-trichloro- and tetrachloroguaiacol
• 2,4-dichloro-, 2,4,6-trichloro-, 2,3,4,6-tetrachloro-, and
pentachlorophenol
t 4-methyl phenol
• Chloroform.
Historical sampling in East Waterway also has been aimed primarily
at determining the spatial extent of contamination in that area and has
not addressed the identification of the sources of that contamination.
Additional data are needed to better define the impact of specific sources
on the receiving environment in East Waterway.
South Port Gardner and Offshore Port Gardner
Much of the deeper portions of South Port Gardner have been sampled
adequately for selected indicator chemicals (e.g. PAH, PCBs, and selected
metals), but data regarding the distributions of other toxic substances
are lacking. In addition, the South Port Gardner shoreline, the deepwater
area near the effluent discharge points for the Scott pulp mill and the
City of Everett CSO, and the area near Mukilteo where high levels of PAH
have been observed are characterized poorly at present. All of these areas
are close to known sources, including CSOs, storm drains, and industrial
effluents.
10
-------
Although Offshore Port Gardner has been sampled infrequently (Map
3), the available data and the distance of this area from sources indicate
that high levels of toxic substances probably are not a problem. However,
the slope area bordering the Snohomish River Delta may be a depositional
zone for material transported by the river, and hence may accumulate higher
concentrations of toxic substances than those measured in shallower waters.
Snohomish River, the Sloughs and Delta
Insufficient data are available for all river, slough, and delta areas
(no analyses of priority pollutants in sediments are available from Ebey,
Steamboat, and Union Sloughs). The few samples collected from the lower
Snohomish River and the delta have contained elevated concentrations of
some selected indicator substances (i.e., PAH, PCBs, and selected metals).
In addition, pulp mills and other industries, CSOs, storm drains, and landfill
leachate discharge into these areas. The impact of these sources on the
receiving environment has not been evaluated.
Port Gardner Disposal Site
A limited number of samples has been collected near the disposal site
southwest of East Waterway. Of these, only one had a data set acceptable
for evaluating selected organic indicator chemicals (i.e., PAH and PCBs).
This limited database is insufficient to characterize adequately the disposal-
site area.
GENERAL STUDY DESIGN
The objectives of the sediment quality survey are to:
t Determine the kind and extent of chemical contamination
in intertidal and subtidal sediments
11
-------
• Characterize the physical properties of sediments that are
related to contaminant availability, transport pathways,
and engineering aspects of remedial action (e.g., dredging)
• Relate environmental contamination to specific sources
• Relate the kinds and magnitudes of toxic contamination to
observed biological effects.
Relationships among physical and chemical properties of sediments, toxicity
measured in the sediment bioassays, and benthic infaunal community character-
istics will be examined using the results of this survey and the related
benthos/bioassay studies described later.
The sediment quality survey consists of the sampling of surface sediments
(i.e., 0-2 cm) at 57 stations throughout the Everett Harbor study area
and at 3 stations in the reference area. The variables to be measured include
the following:
Target Chemicals
• Bulk sediment concentrations of priority pollutants, hazardous
substances, and miscellaneous compounds
Ancillary Parameters
a Total organic carbon and nitrogen
• Water-soluble sulfides
• Total solids
a Grain-size distribution.
The chemical analyses for toxic chemicals will include EPA priority
pollutants and hazardous substance list compounds. Volatile EPA priority
pollutant compounds will only be measured at stations close to potential
sources. Miscellaneous compounds to be analysed at stations near pulp
12
-------
mill discharges will include the chemicals of concern listed previously
(see Data Gaps and East Waterway above).
Ancillary variables will be analyzed in every surficial sediment sample.
Concentrations of organic contaminants can then be normalized to total
organic carbon values of each sediment sample to account for varying ratios
of organic to inorganic substances among samples. Nitrogen content data
will be useful for determining the origin of organic material. Data on
sulfide content will indicate the potential toxicity of bottom sediments
because of anoxic conditions. Grain-size distributions will allow stratifi-
cation of station comparisons based on the physical characteristics of
samples, and will thereby help distinguish effects of pollutants on benthic
infaunal communities from the effects of physical influences of the habitat
itself.
In the the sediment quality survey, only the upper 2 cm of each sediment
sample will be collected and analyzed (Tetra Tech 1986d). At undisturbed
sites, the surface sediments are expected to represent the most recent
contaminant profiles. Because the surface sediment layer also is the most
biologically active zone of the sediments, contaminant concentrations in
surface sediments are of most interest for relating contamination to biological
uptake, bioaccumulation, and other associated effects. Future collection
and analysis of sediment cores may be necessary to define the depth of
problem sediments before an area is subjected to sediment remedial action.
STATION LOCATIONS
Fifty-six subtidal and four intertidal sites will be established
as part of the overall sampling program (Map 11). The majority of the
stations will be located at depths of 30 ft where possible. The 30-ft
depth is shallow enough to be within the potential influence of most contam-
inant sources, yet deep enough to be beyond the influence of sunface waves.
Station locations reflect the following objectives of the sampling plan:
to fill data gaps, to confirm suspected toxic "hot-spots" near major sources,
and to verify questionable data.
13
-------
The majority of subtidal stations are located in shallow nearshore
areas for the following reasons:
• An evaluation of recent studies in the Port Gardner/Everett
project area showed a major data gap for the shallow, nearshore
environment
• Most pollutant sources discharge directly to nearshore areas
• The nearshore environment has the highest incidence of human
contact with toxicants entering Port Gardner, and the greatest
probable effects
• Previous studies in Elliott Bay showed that a major source
of contaminants (Denny Way CSO) had the greatest impact
at depths less than 100 ft (Armstrong et al. 1978; Romberg
et al. 1984)
• For many fish and invertebrate species, the nearshore environment
is prime habitat for foraging, reproduction, and juvenile
development (nursery grounds)
• Feasible remedial actions are most effective in the nearshore
environment.
The breakdown of stations by area within the Port Gardner/Everett
project boundaries is listed below:
Subtidal Intertidal
1. East Waterway 15 0
2. South Port Gardner 11 4
3. Offshore Port Gardner 7
4. Snohomish River Delta 3
14
-------
5. Snohomish River 8 0
6. Port Gardner Disposal Site 0
7. Ebey Slough 3 0
8. Steamboat Slough 6 0
9. Union Slough 0 0
Reference area 3 0
TOTAL 56 4
East Waterway
Fifteen subtidal stations are located in East Waterway. Five stations
are located near active CSOs, storm drains, or industrial outfalls. Seven
stations arelocated at varying distances from these sources to evaluate
contamination gradients. Three stations are located near a potential historical
source of toxic contaminants, the former Weyerhaeuser pulp mill. The main
rationale for station placement is to quantify the degree of contamination
and the magnitude of biological effects in relation to potential sources
of contaminants.
South Port Gardner
Eleven subtidal and four intertidal stations are located in this area.
Subtidal stations occur near the Mukilteo sewage treatment plant, the Mukilteo
fuel facility, Japanese Gulch, Powder Mill Gulch, and Pigeon Creek #1 and
#2. The South Port Gardner shoreline is an area with multiple storm drains,
CSOs, and industrial discharges of which little is known. The proposed
station locations represents the major known point sources for this area
and will enable a comparison of the potential impacts of these sources.
Intertidal sites are located shoreward from several of these subtidal stations
to determine contaminant levels in sediments at public access areas and
beaches.
15
-------
Offshore Port Gardner
Seven subtidal stations are located in this area. All of them are
clustered around the Scott deepwater outfall to evaluate potential contamination
and biological effects related to that discharge.
Snohomish River Delta
Three subtidal stations are located on the river delta to provide
information on levels of contamination in sediments at stations removed
from known sources. One station is located near the mouth of Steamboat
and Ebey Sloughs downstream from the Tulalip landfill and the Weyerhaeuser
lagoon discharge. The other two stations are located 1 and 2 km west of
East Waterway and the mouth of the Snohomish River.
Snohomish River
Eight subtidal stations are located within the Snohomish River downstream
of the 1-5 bridge. One station is positioned near a major CSO discharge
into the Everett Marina Basin. Three stations are located off the historical
discharge from the Weyerhaeuser kraft mill. One station is located mid-channel
off the Everett Wastewater Treatment Plant outfall. One station is located
near CSOs 016 and 017 below the 1-5 bridge. Finally, two stations are
located at mid-channel near the mouth of the river. These latter two stations
are located away from known sources and will be used to evaluate the integrated
potential effects from upstream sources of contaminants.
Port Gardner Disposal Site
The disposal site will be evaluated as part of the Puget Sound Dredged
Disposal Analysis (PSDDA) program. If an initial evaluation indicates
that the site is suitable for future disposal operations, then further
sampling and analysis of the site may be specified as part of PSDDA. Therefore,
sampling of the disposal site will not be conducted as part of the Everett
Harbor Action Program.
16
-------
Ebey Slough
Three subtital stations are located in the main channel of Ebey Slough.
All of these stations are positioned to evaluate levels of contamination
and biological effects potentially associated with leachate from the Tulalip
landfill.
Steamboat Slough
Six subtidal stations are located in the main channel of Steamboat
Slough. One station is located near the Tulalip landfill, whereas the
remaining stations are located near the discharge from the Weyerhauser
kraft mill lagoon.
Union Slough
No sampling is proposed in Union Slough. Because there are no known
sources of pollutants in this area, problems are not expected.
SAMPLING METHODS, PROCESSING, AND ANALYSES
Sediment samples will be collected with a modified O.l-m^ van Veen
grab according to protocols recommended under the Puget Sound Estuary Program
(Tetra Tech 1986d). Each station will be located using the navigation
techniques discussed earlier. Before each sample is taken, vessel position
will be visually rechecked (range alignments) and necessary adjustments
will be made. In response to variability of substrates in the study area,
the field supervisor may use a series of grabs at the same station to obtain
an acceptable depth of grab penetration.
If several attempts to sample a station are unsuccessful, another
nearby station meeting similar sampling needs will be selected and documented.
Standardized data (e.g., collection date, time, station location, depth,
and replicate number) will be recorded at each station.
17
-------
Once onboard, the sample will be held in a vertical position by blocks
and the overlying water carefully drained off by an aspirator hooked to
the ship's hose. The subsamples for volatile organic analyses will be
taken first by placing 40-cm3 glass vials (duplicates) at the undisturbed
sediment surface and filling them using a stainless steel spatula. No
air space will remain in the vials. For the remainder of the subsamples,
aliquots will be taken from a composite sample. The upper 2 cm of sediment
away from the edge of the grab will be removed carefully with a stainless
steel spatula, transferred to a clean stainless steel bowl, and homogenized
by stirring with a glass rod. Aliquots will be collected as follows:
• 500 cm3 will be transferred to a precleaned glass jar with
teflon cap liners (for organic chemical analysis)
• 125 cm3 will be transferred to a precleaned glass jar (for
metals analysis)
• 100 cm3 will be transferred to a Whirl-pak bag (for grain-size
analysis)
• 1,500 cm3 will be transferred to precleaned glass jars (for
bioassays)
• 500 cm3 will be transferred to a precleaned glass jar with
teflon cap liner (for archival storage).
Precleaned (solvent-rinsed and muffle-furnaced) stainless steel bowls
will be brought aboard with replicate solvent-washed spatulas to provide
spares in the event of loss. Bowls will be of adequate size for compositing
samples. Between samples, the bowls will be washed with site water to
remove all residual particulates, washed with pesticide-grade methanol
and pesticide-grade dichloromethane, and then covered with aluminum foil.
Spatulas and stirring rods also will be rinsed with site water, rinsed
with solvent, and wrapped in aluminum foil.
18
-------
Intertidal samples will be collected from shore using a stainless
steel "cookie-cutter" and spatula. Otherwise, intertidal and subtidal
sediment samples will be processed and analyzed in similar fashion to the
above.
In the laboratory, analytical chemistry methods will follow procedures
of the Puget Sound Estuary Program and the U.S. EPA 301 (h) program (Tetra
Tech 1986a) or comparable methods (see Quality Assurance Project Plan,
Tetra Tech 1986b).
19
-------
SEDIMENT BIOASSAYS AND BENTHIC MACROINVERTEBRATE COMMUNITIES
The present study design includes analysis of sediment toxicity (bioassays)
and benthic macroinvertebrate communities at 27 of the subtidal sites chosen
for sediment chemistry (Maps 11 and 12). Sediment bioassays are also planned
for the four intertidal chemistry stations. Station locations for acceptable
data sets from previous studies are shown in Maps 5-8 to allow comparisons
with proposed sampling stations shown in Map 12. The detailed study design
for these components is presented following a summary of data gaps.
DATA GAPS
Benthic Infauna
Major data gaps exist for characterizing benthic infaunal community
structure throughout most of the Everett Harbor study area (Maps 5 and 6).
The main exception is the East Waterway where two recent projects sponsored
by the U.S. Navy have provided adequate spatial coverage. The following
sections present the major data gaps for benthic community structure in
the remaining areas:
• No benthic infauna data for Ebey, Steamboat, and Union Sloughs
are available
• Limited data on benthic infauna are available for the Snohomish
River. Only two stations within the river have been sampled
for determination of species composition and abundance of
benthic infauna
• South Port Gardner lacks adequate data on benthic infauna
20
-------
t Benthic infauna of the Port Gardner Disposal Site have not
been characterized adequately.
Sediment Bioassa.ys
Almost all of the available information on sediment toxicity in the
Everett Harbor study area is for East Waterway. Only three stations outside
this area have acceptable data (Maps 7 and 8). The major gaps in the existing
sediment toxicity data are the following:
• For the East Waterway, the amphipod data are adequate, but
the oyster data are for frozen sediments
• The information for Offshore Port Gardner is extremely limited,
being based on three stations near the mouth of the East
Waterway
• Acceptable sediment-bioassay data are not available for
remaining areas.
GENERAL STUDY DESIGN
The benthic ecology study consists of an assessment of benthic infaunal
communities and sediment toxicity. The main objectives of this study are
to:
• Determine the abundance and distribution of benthic macroinvertebrates
• Determine the toxicity of sediments to a representative
sensitive species
t Relate sediment contamination to biological effects (i.e.,
community structure of benthic infauna and bioassay responses)
21
-------
• Rank areas and contaminants with respect to environmental
impacts.
For most areas, amphipod bioassays are recommended. If sediments
with salinities less than 15 ppt are encounted, other tests may be needed.
The amphipod sediment bioassay measures short-term (i.e. 10-day) response
to bioavailable contaminants and provides an index to estimate effects
on sensitive indigenous organisms by integrating physical, chemical, and
biological aspects of environmental contamination. The benthic infaunal
assessment indicates the ultimate, long-term effects of sediment contamination
at the community level. In addition, benthic infaunal communities and
bioassay data in selected nearshore areas will fill major data gaps for
the project area.
Benthic Infauna
The variables recommended for the benthic infaunal study are the following:
• Total abundance
• Abundances of higher taxa, e.g.,
Polychaeta
Mollusca
Crustacea
Amphipoda
• Species abundances
• Species richness
22
-------
t Species dominance
• Species composition/similarity.
Using these parameters, spatial patterns in biological responses to pollution
can be defined and the relative degree of response at each site can be
estimated.
Total abundance (of all species) and abundances of higher taxa (e.g.,
Echinodermata, Amphipoda, Polychaeta, Gastropoda) will be determined to
assess gross disturbances of benthic communities. Comparisons of community
characteristics among study areas based on analyses of higher taxa will
provide input to site ranking for the Decision Criteria. Statistical comparisons
of data from each area or individual station with reference conditions
will establish a quantitative basis for describing the presence, magnitude,
and spatial extent of biological responses to contamination.
Species richness and dominance and the abundances of indicator species
can be used directly to analyze community properties in relation to site
characteristics. Using numerical clustering techniques, the entire data
set on species abundances can be reduced to an interpretabl e form, whereby
groups of stations are identified on the basis of similarities in their
species composition and relative species abundances (Boesch 1977). Multiple
regression, discriminant analysis, and other multivariate techniques may
be used to relate station-group membership (defined by infaunal community
characteristics) to site characteristics, such as grain-size composition,
depth, organic carbon content, and chemical concentrations. Discrimination
among the potential causes of observed alterations in infaunal communities
will address the importance of conventional physical-chemical parameters
relative to contamination levels.
Because of the high degree of spatial variability in benthic community
characteristics, it is necessary to analyze a sufficient number of replicate
samples. In previous studies of Puget Sound and elsewhere, a minimum of
four to six replicate 0.1-m2 van Veen grabs has been recommended (Holme
23
-------
and Mclntyre 1971; Lie 1968). A total of five 0.1-m2 replicate samples
is usually adequate for most impact assessment work. Lie's analysis of
species area curves showed 75-85 percent of the total species at a site
could be found in five replicates (Lie 1968). Fewer replicate samples
were adequate to characterize the composition of the dominant species assemb-
lages.
A power analysis of replicate infaunal samples from outer Elliott
Bay and the Seahurst area was performed using total abundance, total number
of taxa, echinoderm abundance, and amphipod abundance (data from METRO
Seahurst Baseline Study). The power analyses indicate that five replicate
O.l-m2 samples are adequate for statistically detecting differences in
total number of taxa (and possibly total abundance and amphipod abundance)
among sites. At this level of replication, species-level identifications
are desirable for characterizing species richness as well as for performing
numerical classification analyses. Although use of five replicates probably
has limited value for detecting differences in individual species abundances
(or abundances of rare higher taxa) among sites, increasing the number
of replicates further does not increase statistical sensitivity sufficiently
to justify higher costs.
Sediment Bioassays
The amphipod sediment bioassay will be conducted according to Swartz
et al. (1985), as modified by the Puget Sound Estuary Program (Tetra Tech
and E.V.S. Consultants 1986).
The variables to be measured during the sediment toxicity bioassays
are:
0 Acute (i.e., 10-day) mortality of amphipods (Rhepoxynius
abronius)
• Sublethal effects on R. abronius (moribund, emergence)
24
-------
• Temperature
• pH of sediment
0 Salinity (interstitial and overlying seawater)
• Dissolved oxygen.
Measurement of physical and chemical variables during the bioassays provides
QA/QC and ancillary data for interpretation of results.
STATION LOCATIONS
The locations of proposed sampling stations for benthic infauna and
bioassays in Port Gardner and the Snohomish River system are shown on Map 12.
Sediment chemistry (Map 11) will be evaluated at all stations sampled for
benthic infauna and/or sediment bioassays. In the project area, sediment
for toxicity bioassays will be collected at both intertidal and subtidal
sites, whereas benthic infauna will be characterized only at subtidal sites.
Sampling and analysis of benthic infauna at intertidal sites is not recoirmended
for several reasons. First, an intertidal infaunal survey would require
a major effort because of the diversity of substrate types within the study
area. Second, comparisons among project study areas would require a substantial
sampling effort at a reference site to characterize "background" conditions
for a variety of substrate types. Third, relatively few impact assessment
studies have included intertidal infauna in the past. Fourth, knowledge
about individual species responses is limited, interpretation of the results
would be difficult.
Benthic samples will not be collected at the subtidal biological station
at the Scott deepwater outfall because reference data will not be available
for benthic infaunal communities at that depth. The three benthic stations
in the upper Snohomish River will be sampled, but will not be analyzed
unless adequate reference conditions can be found.
25
-------
The rationale for placement of biological stations is related to that
of the sediment quality studies discussed earlier. In particular, most
of the biological stations are positioned near contaminant sources within
the nearshore areas to allow analysis of gradients in sediment toxicity
and benthic community structure.
SAMPLING METHODS AND SAMPLE PROCESSING
Benthic infauna will be sampled and analyzed according to protocols
recommended under the Puget Sound Estuary Program (Tetra Tech 1986g).
A 0.1-m2 modified van Veen grab will be used to collect sediment for chemical
and benthic infaunal analyses and for toxicity bioassays. At each station,
replicate samples will be taken for analysis of benthic infauna, sediment
chemistry, and bioassays. To avoid disturbance and loss of benthic organisms,
samples for benthic infauna analyses will not be obtained by subsampling
grab samples used for other analyses. Eight to ten sequential grabs will
be made, with alternate grabs for benthos and chemistry/bioassays. Aliquots
of the upper 2 cm from all replicate chemistry/bioassay grabs will be composited
to form a single sample per station. Chemical and physical properties
of the sediments and bioassay responses will thus be measured from a single
composite of all replicate samples collected at a given station.
Because the high cost of chemical analyses and bioassays limits the
number of these measurements that can be made for in replicate samples
in the QA/QC program, analyses of replicate grabs is not desirable. It
is preferable to perform bioassays and chemical analyses on composite sediment
samples to characterize average toxicity at a site. After a benthic sample
is collected, the sample will be washed on a 1.0-mm screen. Mesh of this
size has been used in most previous studies of benthic infauna in Puget
Sound. Samples will be transferred to a container and preserved with 10
percent buffered formalin.
Sediment samples obtained for bioassays will be placed in clean jars
following the homogenizing procedure. Samples will be placed immediately
26
-------
in the dark on ice, transported to the laboratory, refrigerated (4° C),
and then assayed within 14 days of collection.
LABORATORY PROCEDURES
Benthic Infauna
After sitting at least 24 h in fixative, infaunal samples will be
washed on a 0.5-mm screen, transferred to glass jars, and covered with
70 percent ethanol. Using a dissecting microscope, organisms will be removed
from the sediment and sorted in to major taxonomic categories (e.g., Polychaeta,
Oligochaeta, Pelecypoda, Gastropoda, Amphipoda, Isopoda). Specimens from
a given sample and taxonomic group will be placed in separate vials. All
benthic organisms will be identified to species, if possible, or to the
lowest taxon practical.
Benthic samples will be analysed sequentially according to salinity
of the habitat in the Snohomish River estuary and sloughs. Samples from
the most saline areas will be analyzed before samples from less saline
environments. Benthic community structure will not be analyzed for samples
taken upstream from stations where the results indicate a freshwater benthic
community.
QA/QC procedures will follow the Quality Assurance Project Plan (Tetra
Tech 1986b). A reference collection of species identified during the study
will be compiled and archived at U.S. EPA Region X.
Sediment Bioassays
Rhepoxynius abronius will be collected from West Beach on Whidbey
Island, transported to the laboratory, and incubated in their native sediments
for a week prior to use in the assay. During this period of acclimation,
temperature and salinity will be changed gradually at rates no greater
than 1° C and 2 ppt per day until the bioassay conditions of 15° C and
27
-------
25 ppt are attained. Five replicate assays will be performed in 1-1 glass
beakers containing 2 cm (weighted) of test sediment.
Prior to initiation of the bioassay, sediment samples will be mixed
within their storage containers, and pore water will be included in the
final assay of bulk sediments. Once sediment samples are placed in the
beakers, 750 ml of filtered 25-ppt seawater (1 ug, nominal filter diameter)
will be layered onto the sediments and the resultant suspended particulate
matter allowed to settle. Twenty amphipods will then be added to each
replicate beaker and the water overlying the sediments agitated by gentle
bubbling with scrubbed (oil-free), water-saturated air. Bioassays will
be conducted under continuous illumination.
Following 10-day exposure to the test sediments, bioassays will be
terminated by sieving beaker contents through a 0.5-mm screen. Numbers
of surviving amphipods will be counted as those capable of discernible
movement (i.e., pleopod streaming) under a light microscope. At this time,
moribund animals will be identified in a separate assay of burial response
(Swartz et al. 1984).
Appropriate positive (clean sediments) and negative (spiked sediments)
controls will be performed in addition to assays of sediment samples from
the study area. Both organic and inorganic contaminants will be used in
separate series of control experiments.
Sediments from the Snohomish River estuary may be subjected to wide
ranges in salinity depending on the river discharge and tidal pumping.
Interstitial salinities between 15 and 24 ppt will be adjusted upward to
25 ppt. The amphipod bioassay will not be conducted on samples having
an interstitial salinity less than 15 ppt.
28
-------
BIOACCUMULATION AND PATHOLOGY
Because of the potential relationship between bioaccumulation of toxic
substances and prevalence of pathological conditions, these aspects of
the study design are discussed together in this section. Tissue concentrations
of target chemicals provide a measure of contamination of biota, whereas
pathological analyses may indicate sublethal responses of organisms to
chronic exposure to toxic chemicals.
English sole (Parophrys vetulus) was selected as the target fish species
for bioaccumulation and pathology analyses for several reasons. First,
this species is abundant and widespread throughout the Everett Harbor study
area, increasing the probability that adequate sample sizes can be obtained
at all study sites. Second, English sole live in close contact with bottom
sediments, prey mainly on small benthic infauna, and exhibit high levels
of tissue contamination and disease in urbanized areas of Puget Sound (e.g.,
Malins et al. 1984; Tetra Tech 1985a,b). It is therefore likely that this
species is being influenced by contamination of bottom sediments. Finally,
because selected bottomfish species are captured and consumed by some recrea-
tional fishermen (McCallum 1985), English sole are indicative of a food-
web pathway through which contaminants can move from sediments to humans.
According to a recent angler survey in areas of the East Waterway and lower
Snohomish River, however, English sole is not eaten by a large number of
recreational anglers in Everett (McCallum 1985).
Dungeness crab (Cancer magister) was chosen as the representative
invertebrate species primarily because it has shown a tendency to accumulate
contaminants in tissues that are consumed by humans. This species is
sufficiently abundant throughout the study area to allow statistical comparisons
among all study sites. Moreover, Dungeness crabs are an important recreational
and commercial fisheries resource in the Everett area (McCallum 1985; Shapiro
and Associates 1985).
29
-------
DATA GAPS
Bioaccumulation
Data on bioaccumulation of toxic chemicals in edible tissues of indigenous
marine organisms from the study area are available only from one study
by Washington Department of Ecology (Cunningham 1982). Concentrations
of priority pollutants were analyzed in dorsal muscle of rock sole (Lepidopsetta
bilineata) and English sole from five stations in three areas: East Waterway,
lower Snohomish River, and a reference site on the southwest side of Gedney
Island (Maps 9 and 10). Data gaps exist for all other study areas. Edible
muscle tissue of Dungeness crab has not been analyzed for toxic contamination
in the project area. Therefore, the lack of data for target bioaccumulation
variables represents a major data gap.
Pathology
All historical fish pathology data were collected from the East Waterway,
the Snohomish River, the Mukilteo-Everett shoreline, and the area immediately
adjacent to the mouth of the Snohomish River. Major data gaps for fish
pathology include the following areas:
• Parts of South Port Gardner
• Offshore Port Gardner
• The Snohomish River Delta
• Parts of the Snohomish River
• The Port Gardner Disposal Site
0 Ebey Slough
30
-------
• Steamboat Slough
• Union Slough.
GENERAL STUDY DESIGN
The primary objectives of this component are to:
• Determine levels of tissue contamination and frequencies
of pathological disorders in representative fish species
in the Everett Harbor study area
t Compare the level of tissue contamination and prevalence
of disorders among areas
• Relate contamination and disease of organisms to sediment
contamination. Emphasis is placed on obtaining data suitable
for statistical analysis. Results of this study will allow
ranking of areas based on degree of tissue contamination
and disease, identification of local "hot spots", and evaluation
of risk to public health from consumption of contaminated
organisms.
The variables to be measured during the bioaccumulation and pathology
study are:
Chemical concentrations in fish and crab muscle
• Pesticides and PCBs
t Mercury
• Total extractable lipid material
31
-------
Pathological abnormalities in fishes and invertebrates
• External abnormalities for all biota (e.g., lesions, epidermal
papillomas, fin erosion, parasites)
• Internal abnormalities for English sole
• Selected liver lesions for English sole, primarily:
- Neoplasms
- Preneoplasms
- Megalocytic hepatosis
- Nuclear pleomorphism
- Hepatocellular regeneration
- Excessive melanin macrophage centers
Ancillary parameters
• Individual English sole
- Length of all English sole
- Weight, sex, and age of those fish subsampled for histo-
pathological analysis
• Individual Dungeness crabs
- Weight
- Width
- Sex
• Species composition (numerical) of trawl samples.
Chemical analysis of edible portions of target species will allow estimation
of potential human health hazard. PCBs and pesticides will be analyzed
because these substances accummulate in muscle tissue of marine organisms
32
-------
and are a potential threat to public health. Metals other than mercury
will not be analyzed in fish muscle samples because previous data (e.g.,
Cunningham 1982) indicate that no problem exists.
Analyses of contaminants in fish livers is not recommended for this
study because:
• Compounds such as PAH that have been implicated as one potential
cause of liver lesions are detected infrequently in liver
samples using standard techniques
t For PCBs, there may be little relationship between contaminant
concentration in the liver and the prevalence of liver lesions
(Tetra Tech 1985a)
• The total mass of a contaminant in the liver is generally
less than the total mass of that contaminant in edible muscle
tissue (Tetra Tech 1985a). Even for human subpopulations
that consume fish livers, the health hazard associated with
liver ingestion is generally less than that associated with
muscle (fillet) ingestion
• Because of the small mass of liver tissue (typically less
than 2 g), compositing of individual samples would be necessary
for chemical analyses
• Detection limits are generally high even when samples are
composited because of the high lipid content of livers.
The liver is singled out for histopathological analyses because it
is the organ most heavily afflicted with pathological disorders (e.g.,
Mai ins et al. 1980, 1982).. To enhance study efficiency, pathological analysis
of livers will be restricted to six types of idiopathic lesions. These
include hepatic neoplasms, preneoplastic nodules, megalocytic hepatosis,
nuclear pleomorphism, hepatocellular regeneration, and excessive melanin
33
-------
macrophage centers. These disorders are wel1-defined lesions that are
likely to be prevalent enough in the study area to ensure adequate statistical
power of the data analyses. Although the causes of these lesions in field-
caught specimens have not been determined, morphologically similar lesions
have been induced in laboratory mammals and fishes by exposure to toxic
chemicals (Maiins et al. 1984).
Pathological and contaminant analyses will be biased toward larger
English sole (i.e., larger than 230 mm total length, or at least 3 years
old) for two reasons. First, larger fish are the ones most likely to be
retained and consumed by recreational fishermen and therefore pose the
greatest threat to public health if their edible tissue is contaminated.
Second, prevalence of several pathological disorders in English sole livers
increases with age (Malins et al. 1982; McCain et al. 1982; Tetra Tech
1985a). Biasing samples toward larger (i.e., older) fish will ensure that
the study focuses on that portion of the English sole population most likely
to show signs of stress (i.e., lesions). If sufficient samples sizes can
be obtained, an upper size limit (e.g., 300 mm) will also be set for fish
used in the pathology study.
Ancillary data (weight, length, sex, and age) will be collected for
those English sole subsampled for histopathological analysis. Weight-length
relationships for each sex can serve as "condition" indices (e.g., for
comparisons among sites). Length of all remaining English sole will also
be measured. Species composition of each catch will be determined and
these data will be used to characterize and compare fish assemblages.
Sample Sizes
To determine the desirable sample sizes for pathological analyses
of English sole livers, 2x2 contingency analysis was conducted on three
sets of data (Table 1). The question asked was: "Given a certain background
level of disease (i.e., 0, 5, and 10 percent) at what point does an increase
in sample size lead to a negligible improvement (i.e., <2.0 percent) in
the ability to statistically discriminate an elevated level of disease?"
34
-------
TABLE 1 . DETERMINATION OF MINIMUM DETECTION LEVELS FOR ELEVATED
INCIDENCE OF DISEASE GIVEN 10 SAMPLE SIZES AND THREE BACKGROUND
LEVELS OF DISEASE*
Sampl e
Size
20
40
60
80
100
120
140
160
180
200
0 Percent
tf> Dc
20.0
10.0
6.7
5.0
4.0
3.3
2.9
2.5
2.2
2.0
10.0
3.3
1.7
1.0
0.7
0.4
0.4
0.3
0.2
Background Level
s of Disease
5 Percent
% D
30.0
20.0
16.7
15.0
13.0
12.5
12.1
11.3
10.6
10.5
10.0
3.3
1.7
2.0
0.5
0.4
0.8
0.7
0.1
10 Percent
% D
40.0
27.5
23.3
21.3
20.0
19.2
18.6
18.1
17.2
17.0
12.5
4.2
2.0
1.3
0.8
0.6
0.5
0.9
0.2
a Comparisons were made using a 2x2 contingency formulation and the chi-
square criterion.
b Minimum level of disease that Is significantly different (P<0.05) from
background levels.
c Difference in minimum detection levels between two consecutive sample
sizes (i.e., improvement of discriminatory ability).
-------
Results showed that for all three background levels, discriminatory improvement
dropped below 2.0 percent when sample size exceeded 60. Based on experience
in Commencement Bay, a minimum sample size of 60 English sole per station
will allow reasonable discriminatory ability after comparisonwise error
rates are adjusted to compensate for multiple comparisons with the reference
area. Sixty fish will therefore be used for pathological analysis at each
trawl station.
For contaminant analysis of edible tissue in English sole, five fish
from each study site will be used. This sample size is a balance between
analytical costs and even representation across all stations. Gahler et
al. (1982) and Tetra Tech (1985a) used the same sample size to compare
contaminant levels in muscles of English sole between Hylebos and City
Waterways in Commencement Bay and reference sites. Tissue levels of PCBs
were relatively high in the waterways and could be discriminated from those
at the reference site (P<0.05, Mann-Whitney U-test). However, levels of
DDT were only slightly elevated in the waterways and could be discriminated
from background levels only at City Waterway. These results suggest that
a sample size of five may be adequate for discriminating large differences
between contaminated and reference sites (e.g., 100-150 percent of the
grand mean among stations), but may be insufficient for discriminating
smaller differences.
For bioaccumulation in Dungeness crabs, a composite of muscle tissue
samples from 5-10 crabs will be analyzed from each transect. The number
of individual samples in the composite will be the same for all transects.
Results will allow risk estimates to be made for human consumption of contam-
inated crabs. To provide an estimate of variance for QA/QC purposes, three
composite samples will be analyzed from the East Waterway. However, because
estimates of variance will not be available for all sites, statistical
comparisons between transects will not be possible.
35
-------
Sampling Times
To maximize sample sizes and thereby enhance the ability to discriminate
spatial patterns of contamination and disease, all sampling will be conducted
during a single season. Sampling efficiency can be maximized by sampling
between July and September. Because larger fish migrate into the nearshore
zone to feed during this period, catch rates of fish larger than 230 mm
reach an annual peak, and fewer trawl samples should be needed to obtain
required sample sizes.
A second reason to sample English sole between July and September
is that fish are rapidly replenishing lipid reserves following winter fasting
and subsequent spawning (review in Roff 1982). Tissue concentrations of
lipophilic contaminants (e.g., chlorinated hydrocarbons) may therefore
reach an annual peak (i.e., worst-case scenario) during this period. Finally,
because most recreational fishing presumably occurs during spring and summer,
determination of contaminant levels in edible tissue during this period
is probably the most meaningful method of assessing risk to public health
from consumption of contaminated organisms. Late August has been selected
as the optimal sampling period.
STATION LOCATIONS
It is recommended that 11 transects be sampled in the Everett Harbor
study area to evaluate bioaccumulation and fish pathology (Map 12). In
addition, a single transect will be located in the Port Susan reference
area. Most of the transects in the Everett Harbor study area are located
near known or suspected sources of contamination. In the East Waterway
and the Snohomish River, transects will run parallel to the longitudinal
axis of each water body. In other parts of the project area, transects
will be positioned along the 30-ft isobath to coincide with the sampling
depth for sediment chemistry, sediment bioassays, and benthic infauna.
Trawl locations are described below with respect to the specific areas
defined earlier (see above, Figure 1).
36
-------
East Waterway—One transect is recommended for East Waterway. Although
historical data exist for this area, new data will allow interstudy comparisons
to be made and interannual variation to be evaluated.
South Port Gardner—Four trawl transects are recommended for South
Port Gardner. One is located directly offshore from the Mukilteo sewage
treatment plant. A second transect is located off the fuel storage tanks
at Mukilteo. A third transect is located offshore from Powder Mill Gulch.
Finally, a fourth transect is located off the former Weyerhauser pulp mill.
Snohomish River Delta—Two transects are recommended for the Snohomish
River Delta. One is located off Mission Beach near the Tulalip sewage
outfall. The other transect is located approximately 1.5 km northwest
of the mouth of the Snohomish River and East Waterway.
Snohomish River—Two transects are recommended for the Snohomish River.
One transect is located near Baker Island, downstream from the discharge
point of Everett's secondary treatment sewage ponds. The second transect
is located near the mouth of the river, off an adjacent industrial area.
Ebey, Steamboat, and Union Sloughs—Two transects are recommended
for the slough system. One transect in Steamboat Slough is located off
the Tulalip landfill. A second transect in Steamboat Slough is located
off the discharge point of Weyerhauser's kraft-mill lagoon system.
SAMPLING METHODS
English sole will be sampled using a 7.6-m (headrope) otter trawl
having a body mesh size of 3.2 cm (stretched) and a cod-end liner mesh
size of 0.8 cm. As this net has been used by other researchers in Puget
Sound (e.g., University of Washington, National Marine Fisheries Services,
Tetra Tech), data collected in the present study will be directly comparable
with results of most past studies. Mearns and Allen (1978) describe the
sampling device and its operation.
37
-------
Trawls will be made at a constant vessel speed of approximately 1.3 m/sec
(2.5 knots) and each transect will extend approximately 400 m (0.25 mi).
Generally, a 5-min haul will cover the required distance, but this may
vary depending upon strength and direction of currents. Transects based
on distance rather than time are recommended to ensure that sampling effort
is standardized. A minimum of one haul will be made at each site. Additional
hauls may be necessary to obtain required sample sizes.
Because trawling in Port Gardner and the Snohomish River may be complicated
by snags and capture of bottom debris, the trawl will include a polypropylene
(i.e., floatable) retrieval line attached to a float at one end and to
the cod end (by shackle) at the other end. This line allows the net to
be pulled in a reverse direction, and generally frees it from snags and
bottom debris without tearing it. Two complete trawl assemblies will be
onboard, including otter boards, bridles, and nets.
Dungeness crabs will be sampled with the trawls and crab pots. Crabs
captured when trawling will be pooled with those from crab pots if required
to obtain the desired number.
SAMPLE PROCESSING
The recommended sample processing scheme is illustrated in Figure 4.
After each trawl sample is brought aboard, the catch will be sorted
into three categories: 1) English sole, 2) Dungeness crabs, and 3) mis-
cellaneous fishes and invertebrates. All organisms will be examined for
grossly visible external abnormalities while being processed.
Five to ten Dungeness crabs will be randomly selected from those caught
at each site, measured (nearest mm, carapace width), weighed (nearest gm,
wet weight), sexed, frozen whole, and stored for later contaminant analysis
of edible tissue. Invertebrates other than Dungeness crabs and fishes
other than English sole will be identified to species and released. If
38
-------
CRAB-POT SAMPLE
TRAWL SAMPLE
CANCRID CRABS
ENGLISH SOLE
COLLECT
ANCILLARY
DATA
MISCELLANEOUS FISHES
AND INVERTEBRATES
SELECT 5 LARGE
FISH AFTER
LIVER REMOVAL
DETERMINE CONTAMINANT
LEVELS IN EDIBLE TISSUE
IN 5 CRABS AND 5 FISH
I
FILLET
SELECT 60 FISH
LARGER THAN
230 MM TL
±
IDENTIFY, COUNT
AND RELEASE
FIX a 1-CM3
SUBSAMPLEin 10%
FORMALIN
I
REMOVE AND
STORE OTOLITHS
EXAMINE FOR
PATHOLOGICAL
DISORDERS
DETERMINE AGES
Figure 4. Sample processing scheme for pathology and bioaccumulation component.
-------
sufficient numbers of crabs are caught in the nets, the crabs in the pots
will be released.
All English sole will be measured (nearest mm, total length). Sixty
fish larger than 230 mm will be selected randomly, and weighed (nearest gm,
wet weight). The body cavity of each individual will then be opened and
the sex will be determined. These fish will then be examined for gross
visible internal abnormalities, and the liver and otoliths (sagitta) of
each specimen removed. If 60 fish cannot be obtained from the initial
trawl sample, additional hauls will be made until the required sample size
is obtained. Otoliths will be stored for later age determination.
After livers and otoliths have been removed, five of the 60 fish larger
than 230 mm will be selected randomly, wrapped in aluminum, and stored
on ice. The whole fish will be returned to the laboratory where fillets
of dorsal muscle will be removed with stainless steel scalpels. The fillets
will be stored frozen in glass jars for later chemical analysis.
From each of the 60 livers, a 1-cm^ subsample will be excised, placed
in 10 percent buffered formalin, and retained for histopathological analysis.
If a liver contains grossly visible abnormalities, the subsample will be
taken at the border between the normal and abnormal tissue and will include
both types of tissue. If no abnormalities are visible, the subsample will
be taken from the center of the liver at its broadest point.
39
-------
DATA MANAGEMENT
All data for the project, including field observations, will be entered
onto pre-formatted data log sheets. The completed sheets will be entered
into the project Data Management System (DMS) in National Oceanographic
Data Center (NODC) formats.
Upon entry of a data set, the scientist who generated the data will
be provided with hard copies of the computer data file, together with basic
data quality analyses (e.g., means, standard deviations, and ranges) of
each parameter. Quality assurance tests for each data set will be conducted
by the appropriate scientist. These outputs will be reviewed to ensure
that accurate data have been entered into the data files.
40
-------
SUMMARY
A summary of spatial coverage and sampling effort for the Everett
Harbor study design is provided in Table 2. The study design has been
developed to provide a comprehensive assessment of contamination and its
effects within the study area. The objective has been to gain as much
information as possible within the constraints of preliminary estimates
of program cost and available funding resources.
41
-------
TABLE 2. SUMMARY OF BASIC STUDY DESIGN
Project Sediment Quality Bioassays3
Subarea Subtidal Intertidal Subtidal Intertidal
East
Waterway
South Port
Gardner
Offshore Port
Gardner
Snohomish
River Delta
Snohomish
River
Ebey Slough
Steamboat
Slough
Port Susanc
Total No.
Stations
Total No.
Samples
15
11
7
3
8
3
6
3
56
56
0 6
4 6
1
0 2
0 5
0 3
0 2
0 3
4 28
4 28
0
4
0
0
0
0
0
4
4
Benthic Fish
Infauna3 Bioaccumulation^ Pathologyb
Subtidal Subtidal Subtidal
6
6
0
2
5
3
2
3
27
135d
1
4
0
2
2
1
2
1
13
80e
1
4
0
2
2
0
2
1
12
720f
a Sediment chemistry will be measured at all stations sampled for benthic infauna and sediment
bioassays.
b Subtidal bioaccumulation and pathology samples will be taken from the same trawls.
c Reference area.
& Five replicate 0.1-m2 van Veen samples per station for benthic infauna.
e Edible muscle tissue from five English sole at each station and a single composite of 5-10
Oungeness crabs at all stations except East Waterway, where 3 composites will be analyzed.
f Sixty English sole livers for histopathological analyses at each station.
-------
REFERENCES
Armstrong, J.W., R.H. Thorn, K.K. Chew et al. 1978. The impact of the Denny
Way combined sewer overflow on the adjacent flora and fauna in Elliott
Bay, Puget Sound, Washington. Municipality of Metropolitan Seattle, Seattle,
WA. 102 pp.
Boesch, D.F. 1977. Application of numerical classification in ecological
investigations of water pollution. EPA-600/3-77-033. U.S. EPA Environmental
Research Laboratory, Corvallis, OR. 125 pp.
Cunningham, D. 10 November 1982. Memo: Assessment of toxic pollutants
in English sole and rock sole in Everett Harbor and Port Gardner. Washington
Department of Ecology, Olympia, WA. 28 pp.
Gahler, A.R., R.L. Arp, and J.M. Cummins. 1982. Chemical contaminants
in edible non-salmonid fish and crabs from Commencement Bay, Washington.
U.S. EPA Environmental Services Division, Seattle, WA. 117 pp.
Holme, N.A., and A.D. Mclntyre. 1971. Methods for the study of marine
benthos. IBP Handbook No. 16. Blackwell Scientific Publications, Oxford,
UK. 334 pp.
Lie, U. 1968. A quantitative study of benthic infauna in Puget Sound,
Washington, U.S.A. in 1963-1964. Fisk. Skr., Sen. Hav. 14:229-556.
Malins, D.C., B.B. McCain, and D.W. Brown. 1980. Chemical contaminants
and biological abnormalities in central and southern Puget Sound. NOAA
Technical Memorandum OMPA-2. National Ocean and Atmospheric Administration,
Boulder, CO. 295 pp.
Malins, D.C., B.B. McCain, D.W. Brown et al. 1982. Chemical contaminants
and abnormalities in fish and invertebrates from Puget Sound. NOAA Technical
Memorandum OMPA-19. National Oceanic and Atmospheric Administration, Boulder,
CO. 168 pp.
Malins, D.C., B.B. McCain, D.W. Brown et al. 1984. Chemical pollutants
in sediments and diseases of bottom-dwelling fish in Puget Sound, Washington.
Environ. Sci. Technol. 18:705-713.
McCain, B.B., M.S. Myers, and U. Varanasi. 1982. Pathology of two species
of flatfish from urban estuaries in Puget Sound. NOAA Northwest and Alaska
Fisheries Center, Seattle, WA. 100 pp.
McCallum, M. 1985. Recreational and subsistence catch and consumption
of seafood from three urban industrial bays of Puget Sound: Port Gardner,
Elliott Bay, and Sinclair Inlet. Washington State Division of Health,
Epidemiology Section, Department of Social and Health Services, Olympia,
Washington. 59 pp.
42
-------
Mearns, A.J., and J.M. Allen. 1978. Use of small otter trawls in coastal
biological surveys. EPA-600/3-78-083. U.S. Environmental Protection Agency,
Con/all is, OR. 33 pp.
Roff, D. 1982. Reproductive strategies in flatfish: a first synthesis.
Can. 0. Fish. Aquat. Sci. 39:1686-1698.
Romberg, G.P., S.P- Pavlou, and E.A. Crecelius. 1984. Presence, distribution,
and fate of toxicants in Puget Sound and Lake Washington. Metro Toxicant
Program Report No. 6A. Toxicant Pretreatment Planning Study Technical
Report Cl. Municipality of Metropolitan Seattle, Seattle, WA. 231 pp.
Shapiro and Associates, Inc. 1985. Draft Environmental Impact Statement
Supplement. Everett Harbor and Snohomish River Navigation Project. Prepared
for U.S. Army Corps of Engineers by Shapiro and Associates, Seattle, WA.
51 pp. + appendices.
Swartz, R.C., W.A. DeBen J.K.P. Jones, J.O. Lamberson, and F.A. Cole.
1985. Phoxocephalid amphipod bioassay for marine sediment toxicity.
pp. 284-307. In: Aquatic Toxicology and Hazard Assessment, Proc. of the
Seventh Annual Symposium. ASTM-STP 854. American Society for Testing
and Materials, Philadelphia, PA.
Tetra Tech. 1985a. Commencement Bay nearshore/tideflats remedial investi-
gation. Tetra Tech, Inc., Bellevue, WA.
Tetra Tech. 1985b. Everett Harbor action plan: initial data summaries
and problem identification. Prepared for U.S. Environmental Protection
Agency, Region X, Office of Puget Sound. Tetra Tech, Inc., Bellevue, WA.
81 pp. + appendix.
Tetra Tech. 1985c. Sampling and analysis design for development of Elliott
Bay toxics action plan. Prepared for U.S. Environmental Protection Agency,
Region X, Office of Puget Sound. Tetra Tech, Inc., Bellevue, WA. 69 pp. +
appendix.
Tetra Tech. 1986a. Analytical methods for U.S. EPA priority pollutants
and 301(h) pesticides in marine and estuarine sediments. Prepared for
Office of Marine and Estuarine Protection, U.S. Environmental Protection
Agency, Washington, DC. Tetra Tech, Inc., Bellevue, WA.
Tetra Tech 1986b. Quality assurance project plan for field investigations
to support development of the Everett Harbor action plan. Prepared for
U.S. Environmental Protection Agency, Region X, Office of Puget Sound.
Tetra Tech, Inc., Bellevue, WA.
Tetra Tech. 1986c. Recommended protocols for conducting fish pathology
studies in Puget Sound. Draft Report. Prepared for U.S. Environmental
Protection Agency, Region X, Office of Puget Sound. Tetra Tech, Inc.,
Bellevue, WA.
43
-------
Tetra Tech. 1986d. Recommended protocols for measuring conventional sediment
variables in Puget Sound. Prepared for U.S. Environmental Protection Agency,
Region X, Office of Puget Sound. Tetra Tech, Inc., Bellevue, WA. 46 pp.
Tetra Tech. 1986e. Recommended protocols for measuring metals in Puget
Sound sediment and tissue samples. Draft Report. Prepared for U.S. Environ-
mental Protection Agency, Region X, Office of Puget Sound. Tetra Tech,
Inc., Bellevue, WA.
Tetra Tech. 1986f. Recommended protocols for measuring organic compounds
in Puget Sound sediment and tissue samples. Draft Report. Prepared for
U.S. Environmental Protection Agency, Region X, Office of Puget Sound.
Tetra Tech, Inc., Bellevue, WA. 55 pp.
Tetra Tech. 1986g. Recommended protocols for sampling and analyzing subtidal
benthic macroinvertebrate assemblages in Puget Sound. Draft Report. Prepared
for U.S. Environmental Protection Agency, Region X, Office of Puget Sound.
Tetra Tech, Inc., Bellevue, WA. 37 pp.
Tetra Tech and E.V.S. Consultants. 1986. Recommended protocols for conducting
laboratory bioassays on Puget Sound sediments. Final Report. Prepared
for U.S. Environmental Protection Agency, Region X, Office of Puget Sound.
Tetra Tech, Inc., Bellevue, WA. 55 pp.
44
-------
EVERETT
•
-------
EVERETT
EAST WATERWAY
SURFACE RUNOFF
CSO
• INDUSTRIAL DISCHARGE - EXISTING
D INDUSTRIAL DISCHARGE HISTORICAL
SCOTT
EVERETT
(WG) WESTERN GEAR
0 250
Contaminant sources and selected industry
locations in East Waterway of Everett Harbor.
-------
1 2
NAUTICAL MILES
KILOMETERS
2 CONTOURS IN FEET
Sediment Chemistry: Sampling stations for selected data
sets in Everett Harbor.
MAPS
• SAMPLING STATION
INTERTIDAL AREAS
-------
EP19-31 ;
• B6-E8 /
EP19-30
30
B6-E14
EAST WATERWAY EP19-29
B8-29 •
B6-E16 EP20-23
EP19-28 •
EVERETT
EP19.26 B6-E18
SAMPLING STATION
250
500
0 250
CONTOURS IN FEET
YARDS
METERS
500
Sediment Chemistry: Sampling stations for selected data
sets in East Waterway of Everett Harbor.
MAP 4
-------
1 2
NAUTICAL MILES
KILOMETERS
2 CONTOURS IN FEET
Benthic Infauna: Sampling stations for selected data
sets in Everett Harbor.
MAPS
• SUBTIDAL SAMPLING STATION
INTERTIDAL AREAS
-------
U5-E2
EAST WATERWAY
• SAMPLING STATION
60
0 250
CONTOURS IN FEET
Benthic Infauna: Sampling stations for selected data
sets in East Waterway of Everett Harbor. MAP 6
-------
1 2
NAUTICAL MILES
KILOMETERS
2 CONTOURS IN FEET
Sediment Bioassay: Sampling stations for selected data
sets in Everett Harbor.
MAP 7
0 OYSTER LARVAE BIOASSAY
4 AMPHIPOD BIOASSAY
SAMPLES WERE COMPOSITED
INTERTIDAL AREAS
-------
30
.-• o *B9'5
U8-E15'4>. CH8-22
U8-E14
EAST- WATERWAY
U8-E16
0 OYSTER LARVAE BIOASSAY
• AMPHIPOD BIOASSAY
SAMPLES WERE COMPOSITED
250
500
YARDS
METERS
0 250
CONTOURS IN FEET
EVERETT
Sediment Bioassays: Sampling stations for selected data
sets in East Waterway of Everett Harbor. MAP 8
-------
EVERETT
FISH TRAWL/PATHOLOGY
BIOACCUMULATION
INTEFITIDAL AREAS
(TRAWL LOCATIONS ARE APPROXIMATE)
Fish Pathology and Fish Bioaccumulation: Sampling
stations for selected data sets in Everett Harbor.
MAP 9
-------
EVERETT
FISH TRAWL/PATHOLOGY
• BIOACCUMULATION
TRAWL LOCATIONS ARE APPROXIMATE
250
500
0 250
CONTOURS IN FEET
YARDS
METERS
500
Fish Pathology and Fish Bioaccumulation: Sampling
stations for selected data sets in East Waterway
of Everett Harbor. MAP 10
-------
NOTE: 3 STATIONS WILL BE SAMPLED
FOR PP + PM IN THE PORT SUSAN
REFERENCE AREA.
Sampling locations for recommended chemical
studies in Everett Harbor.
MAP 11
o PP
• PP-V
® PP + PM
* PP-V+PM
A V + PM
PP - Priority Pollutants
V » Volatile Compounds
PM * Pulpmill Compounds
-------
NOTE: 3 BIOASSAY/BENTHOS STATIONS
AND 1 TRAWL TRANSECTWILL BE
SAMPLED IN THE PORT SUSAN
REFERENCE AREA.
NAUTICAL MILES
CONTOURS IN FEET
® INTERT1DAL BIOASSAY
* SUBTIDAL BIOASSAY
• SUBTIDAL BIOASSAY/BENTHOS
•4 SUBTIDAL PATHOLOGY/BIOACCUMULATION
Sampling locations for recommended biological
studies in Everett Harbor.
------- |