The 1991 State/Federal
Natural Resource Damage Assessment
        and Restoration Plan
    for the Exxon  Valdez Oil Spill

   Volume I: Assessment and Restoration Plan
           Appendices A, B, C
       Mlil

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                                                  April 1991
Dear Reviewer:

This document describes studies proposed to be conducted jointly by
the State of  Alaska and the United States  during  the third year
since the Exxon Valdez oil spill.   The purpose of these studies is
to determine injury to natural resources resulting from that spill.
This  document  also  describes  restoration  planning  activities
proposed for 1991.

The 1991 proposed plan has greatly benefitted by incorporation of
many of the public comments on the  "State/Federal Natural Resources
Damage Assessment Plan and  Restoration  Plan for the Exxon Valdez
Oil Spill, August 1990."  This proposed plan was assembled through
the cooperative efforts of the State of Alaska acting through the
Departments of Fish and Game, Environmental Conservation, Natural
Resources,  and Law,  and the  United States  acting  through the
Federal  Departments  of  Justice,   Agriculture and  Interior,  the
National  Oceanic and Atmospheric Administration,  and  the  U.S.
Environmental Protection Agency.

At this printing an agreement has been  reached between the State
and Federal Trustees, and Exxon, regarding a judicially supervised
settlement of claims.   Ratification of the settlement agreement may
result in modification of plans  and projects currently proposed to
be conducted.

Public comment on this document  will assist  the Trustee Council in
developing future injury assessment and restoration efforts and may
also result in modification of  plans  and projects  proposed to be
conducted  in  1991.    Questions   concerning the   plan  and  its
distribution should be directed  to U.S. Department  of Agriculture,
Forest Service Public Affairs Office (907)   586-8806.
                                           printed on 50% recycled paper

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Comments  should  be received  by  June 3,  1991,  at  the  following
address:

                    Trustee Council
                    P. O. Box 22755
                    Juneau, AK  99802


We appreciate your interest and look  forward to your participation
in this important process.

Sincerely,
Michael A. Barton
Regional Forester
Alaska Region
Forest Service
Department of Agriculture
Charles E. Cole
Attorney General
State of Alaska
Steven Pennoyer
Director
Alaska Region
National Marine Fisheries Service
Carl L. Rosier
Commissioner
Alaska Department of
Fish and Game
John R. Sandor
Commissioner
Alaska Department of
Environmental Conservation
Walter O. Stieglitz
Director
Alaska Region
Fish and Wildlife Service
Department of the Interior

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               VOLUME  I:   THE 1991 STATE/FEDERAL NATURAL RESOURCE
                 DAMAGE ASSESSMENT AND RESTORATION FLAN FOR THE
                  EXXON VALDEZ OIL SPILL  AND APPENDICES A,  B, C
en
en
Q_
                               HEADQUARTERS LIBRARY
                               ENVIRONMENTAL PROTECTION AGENCY
                               WASHINGTON, D.C. 20460

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                        TABLE OF CONTENTS

VOLUME I

INTRODUCTION 	   1

PART I

Injury Determination/Quantification

     Marine Mammal Injury Assessment 	   9
     Terrestrial Mammal Injury Assessment 	  48
     Bird Injury Assessment 	  60
     Fish/Shellfish Injury Assessment 	  87
     Coastal Habitat Injury Assessment 	 175
     Subtidal Injury Assessment 	 186
     Technical Services 	 251
     Archaeological Resources Injury Assessment 	 256

PART II

Peer Reviewers/Chief Scientist 	 259

PART III

Economics 	 260

PART IV

Oil Spill Public Information Support 	 275

PART V

Restoration Planning 	 276

PART VI

Budget 	 283

APPENDICES

A.   Quality Assurance/Quality Control 	 A-l
B.   Histopathology Procedures 	 B-l
C.   Glossary of Terms and Acronyms 	 C-l

VOLUME II

Appendix

D.   Response to Public Comments	 D-l

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                      INTRODUCTION

The  March  24,  1989,  grounding of  the  tanker  Exxon  Valdez  in
Alaska's Prince William Sound caused the largest  oil  spill  in  U.S.
history.   Approximately 11 million gallons  of  North Slope crude
moved through the southwestern portion  of the Sound  and along the
coast of the western Gulf of Alaska (see map,  Fig.  1). The spill
injured  fish,  birds,  mammals,  and a  variety  of  other  forms  of
marine life, habitats, and resources.

The State of Alaska  acting through the  Alaska Departments  of  Fish
and  Game  (ADF&G),   Environmental  Conservation  (ADEC),  and  Law
(Attorney  General),   and  the  United States  acting  through  the
federal  Departments  of  Agriculture  (DOA),  Interior  (DOI)  and
through the National  Oceanic and Atmospheric Administration  (NOAA),
are acting together as Natural Resource Trustees as provided by the
Comprehensive Environmental Response, Compensation,  and Liability
Act  (CERCLA),  the  Clean  Water  Act (CWA),  and  other  state  and
federal authorities.   The  Environmental Protection Agency (EPA)  is
assisting  in  damage  assessment  and is  coordinating the  federal
restoration efforts  with  the State of Alaska.

This plan, which describes the proposed 1991  studies,  continues  or
modifies certain  1989 and 1990 damage  assessment studies.   These
studies  are designed to  determine  the nature and extent  of the
injuries, losses or destruction of resources, and lost uses of the
resources.  These data provide a base for developing  a restoration
plan.

Funds received as the result of litigation or settlement will  be
used to restore,  replace,  or acquire the equivalent of the  injured
natural  resources and services  and to reimburse  agencies  for
relevant costs incurred.   The U.S. Department of Justice and Alaska
Department  of  Law represent  the federal and  state   governments,
respectively, in pursuit  of these claims.

In  1989,   the  Trustees   developed  a  damage  assessment   plan
incorporating 72  studies  in 10 categories.  In  1990, 50  studies
were  undertaken.     The   proposed  1991  damage   assessment plans
incorporates 42 studies in 10 categories.

Damage assessment  is a dynamic  process and it  will continue  to
evolve.  In order to identify studies  that  should be continued,
terminated or new studies that should  be initiated,   the Trustees
considered the extensive public comments on the first two years  of
work and consulted damage assessment investigators,   other agency
scientific  staff,  legal counsel, and  independent outside expert
reviewers.  The studies were evaluated from five perspectives:  (1)
immediate  injury,  (2)  long-term  alteration of  populations,  (3)
sublethal or latent  effects,  (4)  ecosystem-wide  effects,  and (5)
habitat degradation.

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Studies were discontinued  for  a variety of reasons,  such  as the
determination that field work had been completed or that there was
no practicable way to measure injury.  The mere fact that a study
was discontinued does not indicate that the resource was uninjured
by the spill.   Funds are provided to conclude data  analysis and
report preparation for certain studies that are not being continued
in 1991.

The  studies  described  in this plan fall  into  ten  categories:
(1) Marine Mammals,   (2)  Terrestrial Mammals,  (3)  Birds,    (4)
Fish/Shellfish,  (5)  Coastal Habitat,  (6)  Subtidal Habitat,  (7)
Technical  Services   (including  chemistry  and   an   integrated
geographic information system,  complete with mapping)  to support
the resource studies,  (8)  Archaeological  Resources,  (9)  Economic
Studies,   and  (10)  Restoration.   The  cost for  all  activities
described  in  the  1991   State/Federal  Natural  Resource  Damage
Assessment and Restoration Planning for the Exxon Valdez Oil Spill
is approximately $35 million.

Marine Mammal studies include direct  observations of injury (e.g.,
through carcass counts)  as well  as estimates of population effects
based on censuses or pathologic and toxicologic indicators (as is
being undertaken with otters and seals).  In  addition,  the direct
observational data allow for inferences to be made about injuries
to populations.

Terrestrial  mammals  near  the  coast may  have been  exposed  to
hydrocarbons  by breathing  fumes  or eating oiled carcasses  or
vegetation. The studies will determine the  presence of hydrocarbons
in tissues of dead animals and the effects, if any, of oil exposure
on local populations of brown bears and river otters.

The  1991  effort  to  determine injury  to  birds   will   focus  on
seabirds, bald eagles, and waterfowl.  Surveys and censuses, radio
telemetry, and documentation of sublethal and physiological impacts
will  be  used  as  means  to  determine  injury.    The  information
obtained  will  contribute  to  an  understanding  of  mortality,
population changes,  and other  factors  essential  for  the damage
assessment process.  Studies proposed  for  birds  focus on  the
collection of data on survival and reproductive success in relation
to initial and continuing exposure to hydrocarbons and conversion
products.

The Fish/Shellfish studies focus on identifying potential  injury to
the various life stages of fish  and shellfish in areas affected by
the  oil  spill. Species  were selected for  study  based  on their
respective niche  or  overall  importance  within  the  ecosystem,
ability to be sampled, and the existence of an historic data base.

The  Coastal  Habitat study measures  spill-related  changes in the
intertidal and shallow subtidal  zones.  It is designed to document
injury to  resources that rely  on  these habitats,  and  to assess

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damages for the loss of services provided by these habitats.

The  Subtidal  Habitat  studies  determine  the  distribution  and
composition  of  petroleum  hydrocarbons  or  their  environmental
conversion  products  in water,  sediments,  and  living resources.
Information  gathered  on  the  distribution  and  nature  of  the
hydrocarbons and  their conversion products provides  a basis for
documenting exposure and for determining  injury  to resources.  The
combined  results of  the  Coastal Habitat  and Subtidal  Habitat
studies also  form a  basis for  estimating  rates of  recovery of
natural resources and the potential for accelerating recovery.

The Technical  Services  category includes activities that provide
process support or information services to all studies  in the areas
of analytical  chemistry and an integrated geographic information
system, complete with mapping.

Studies on archaeological resources will proceed in two steps:  (1)
inventory, description,  and classification; and  (2) qualitative and
quantitative descriptions and measurements of changes detrimental
to the archaeological resources related to the spill.

The value of lost or injured natural resources, and the goods and
services they  provide humans,  are based  on  results  from economic
studies.    In  this  regard,  damages  forming  the  basis  of  the
Trustees'  claim  against the potentially responsible  parties are
calculated  by  considering  (1)  the  reduction of these  goods and
services,  including intrinsic values, resulting from  the spill, and
(2)  the cost  of restoring these  goods and  services to  their
pre-spill level, replacing them, or acquiring their equivalent.

The restoration planning component describes the strategy and scope
of the restoration process  planned  for the  third  oil  spill year.
Restoration measures will  be implemented as appropriate methods are
identified and funds are available.

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TABLE ONE.  STUDIES AUTHORIZED IN 1989, 1990 AND 1991



X = Initiated or Continued
STUDY
CATEGORY
Marine Mammals
1






Terrestrial





Birds







2
3
4
5
6
7
STUDY TITLE
(MM)
Humpback Whale
Killer Whale
Cetacean Necropsy
Sea Lion
Harbor Seal
Sea Otter Injury
Rehabilitated Sea Otters
Mammals (TM)
1 Sitka Black-Tail Deer
2
3
4
5
6
1
2
3
4
5
6
7
8
Black Bear
River Otter & Mink
Brown Bear
Small Mammals
Mink Reproduction
Beached Bird Survey
Census/Seasonal Distribution
Seabird Colony Surveys
Bald Eagles
Peale's Peregrine Falcons
Marbled Murrelets
Storm Petrels
Black-Legged Kittiwakes
1989
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
1990
X
X

X
X
X
X
X
X
X
X

X
X
X
X
X
X



1991

X

*
X
X
moved to MM 6


X
X

*
X
X
X
X





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TABLE ONE (Con't).  STUDIES AUTHORIZED IN 1989, 1990 AND 1991
STUDY
CATEGORY STUDY TITLE
Birds, continued
9 Pigeon Guillemots
10 Glaucous-winged Gulls
11 Sea Ducks
12 Shorebirds
13 Passerines
14 Exposure North Slope Oil
1989
X
X
X
X
X
X
1990 1991


X X

X

Fish/Shellfish (F/S)
            1  Salmon Spawning Area Injury
            7  Salmon Spawning Area
               Injury, Outside PWS

            8  Egg & Pre-emergent Fry,
               Sampling Outside PWS

            9  Early Marine Salmon
               Injury Outside PWS

           10  Dolly Varden & Sockeye
               Injury, Lower Cook Inlet

           11  Herring Injury

           12  Herring Injury Outside PWS

           13  Clam Injury

           14  Crab Injury

           15  Shrimp Injury
                                   X
2  Eggs/Pre-emergent Fry Sampling  X

3  Coded-wire tagging              X

4  Early Marine Salmon Injury      X

5  Dolly Varden Injury             X

6  Sport Fishing Harvest & Effort  X

                                   X
                                   X


                                   X


                                   X


                                   X

                                   X

                                   X

                                   X

                                   X
X
X
X
X
X
X
X
X
X
X
          X
          X
X  moved to subtidal

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TABLE ONE (Con't).  STUDIES AUTHORIZED IN 1989, 1990 AND  1991
     STUDY
   CATEGORY
STUDY TITLE
1989
1990
1991
Fish/Shellfish, continued
           16  Oyster Injury
           17  Rockfish Injury

           18  Trawl Assessment

           19  Larval Fish Injury

           20  Underwater Observations

           21  Clam Injury Outside PWS

           22  Crab Injury Outside PWS

           23  Rockfish Injury Outside PWS

           24  Dermersal Fish Injury

           25  Scallop Mariculture Injury

           26  Sea Urchin Injury

           27  Sockeye Salmon Overescapement

           28  Run Reconstruction

           29  Life History Modeling

           30  Database Management
Coastal Habitat  (CH)
            1  Intertidal Studies
Air/Water (A/W)
            1  Geographic Extent of
               Oil in Water

            2  Injury to Subtidal Sediments
               and Benthos

            3  Hydrocarbons  in Water

            4  Injury to Deep Water

            5  Injury to Air

            6  Oil Fate and  Toxicity
                                 X

                                 X

                                 X

                                 X

                                 X

                                 X

                                 X

                                 X

                                 X

                                 X

                                 X
                                 X




                                 X



                                 X



                                 X

                                 X

                                 X
            X  moved to subtidal

            X         *
        (combined with F/S 13)

            X

        (combined with F/S 17)

            X  moved to subtidal
                                           X          X

                                           X          X

                                        (combined with F/S 28)

                                           X          X
            X
            X  moved to subtidal


            X  moved to subtidal

         (combined with A/W 2)



            X  moved to subtidal

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TABLE ONE  (Con't).  STUDIES AUTHORIZED IN 1989, 1990 AND 1991

     STUDY
   CATEGORY
STUDY TITLE
1989
1990
1991
Subtidal
            1  Hydrocarbon Exposure, Microbial and
               Meiofaunal Community Effects  (A/W2)

            2  Injury to Benthic Communities  (CH 1 and A/W 2)

            3  Bio-availability and Transport
               of Hydrocarbons (A/W 3)

            4  Sediment Toxicity Bioassays (A/W 6)

            5  Injury to Shrimp (F/S 15)

            6  Injury to Rockfish  (F/S 17)

            7  Injury to Demersal Fish (F/S 24)
Technical Services
            1  Hydrocarbon Analysis

            2  Histopathology

            3  Mapping
Archaeology
            1  Archaeological
Economics
                                 X         X

                                 X         X

                                 X         X



                          Part of Econ 9   X
            1  Commercial Fisheries Losses     X

            2  Fishing Industry Costs          X

            3  Bioeconomic Models              X

            4  Public Land Effects             X

            5  Recreational Losses             X

            6  Subsistence Losses              X

            7  Intrinsic Values                X
                      X


                      X

                      X


                      X

                      X

                      X

                      X



                      X
                      X
                                           X         X

                                       (combined with Econ 1)

                                       (combined with Econ 1)

                                           X

                                           X         X

                                           X         X

                                           X         X

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TABLE ONE (Con't).  STUDIES AUTHORIZED IN 1989, 1990 AND 1991

     STUDY
   CATEGORY   STUDY TITLE	1989	1990	1991

Economics, continued
            8  Research Program Effects        XXX

            9  Archaeological Damage           X
               Quantification

           10  Petroleum Products Price                            X


Restoration Planning                           XXX
*  These studies  are  being funded for the completion  of  data analysis and
final report preparation.

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PART I:  INJURY DETERMINATION/QUANTIFICATION

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                     MARINE  MAMMAL ASSESSMENT

Although the most visible impact of  the EVOS on marine mammals was
the large number  of  dead sea  otters,  other marine mammal species
were potentially injured by the spill,  including Steller sea lions,
harbor seals, killer whales, and humpback whales.

In 1989,  seven  studies were assembled and  implemented  to gather
information  on  injury  to marine  mammals.    Aerial  surveys  for
stranded  cetaceans  were also  conducted.    Additional  data  on
injuries  to   sea  otters  were   gathered  at   the   sea  otter
rehabilitation centers.

In 1990,  most of these studies were continued to further refine the
information documenting injury resulting from the spill.

Three of these studies  will  be continued in 1991 including studies
on killer whales,  harbor  seals  and  sea  otters.   In addition,  the
study on sea lions conducted during 1989 and 1990 will be completed
with final data analysis and report preparation.

In many  cases, the 1989 and 1990 studies have been expanded  and
modified  in response to  knowledge gained  during the  two  years
following the spill, as  well  as,  comments from reviewers and the
public.  The ongoing study on killer whales is intended to provide
information on changes  in killer whale use  of the spill zone,  to
assess long-term impacts, and to corroborate information on injury
to killer whales gathered during the 1989 and 1990 studies.   Data
from  studies  on harbor   seals   will   provide   information  on
toxicological effects  of the  EVOS.   The  sea  otter study  will
continue to look at population effects and possible physiological
and toxicological impacts that could result in  long-term, sublethal
injuries.

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MARINE MAMMAL STUDY NUMBER 2

Study Title:  Assessment of Injuries to Killer Whales in PWS

Lead Agency:  NOAA


                           INTRODUCTION

During the first two years  of  the  killer  whale damage assessment
work, photographs of individual killer whales in PWS were collected
from May to  September  1989 and 1990 to assess the  impact  of the
EVOS on killer  whale  life history and ecology.  In  PWS,  research
vessels traversed over  20,000 nautical miles in search of whales or
while photographing whales,  reflecting 507 days of field research.
This effort represents  the most complete study accomplished to date
on killer whales in PWS.  An unusually high number of killer whales
have been reported missing  from the  PWS area.   The  assessment of
the overall effects of  the EVOS on  killer whale populations in PWS
will be enhanced with photographic evidence that the whales missing
in 1989/1990 are confirmed missing in 1991.

The purpose of  this study  is to  obtain  photographs  of individual
killer whales in PWS from  May  to  late September  1991.  Calves of
the  year will  be  documented.    Photographs  collected will  be
compared to the  Alaskan photographic database for the years 1977 to
1990 to  determine  if  changes  have occurred  in  whale abundance,
seasonal distribution,  continuity of  habitat usage, pod integrity,
and mortality or natality rates.  Results of  this  research will aid
in the determination of the extent  of displacement or reduction in
numbers of killer whales as a result of the EVOS.


                            OBJECTIVES

A.   To count the number and individually identify killer whales
     within PWS.

B.   To test the hypothesis that killer whale distribution within
     PWS and adjacent waters  is  similar  to that  reported for
     previous years (1984-1990).

C.   To test the hypothesis that pre- and post-spill killer whale
     pod structure and integrity have remained constant.

D.   To test the hypothesis that killer whale natality rates within
     PWS have not changed since the  EVOS.

E.   To  test the  hypothesis  that  killer whale  mortality rates
     within PWS have not changed since the EVOS.
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                             METHODS

Personnel  from  the  National  Marine Mammal  Laboratory  (NMML),
Seattle,  Washington  (Alaska  Fisheries  Science  Center,  NMFS/
NOAA/DOC) will  develop  and coordinate all killer  whale research
activities  associated  with the  Exxon Valdez damage  assessment.
Field studies will be conducted by contractors that have recognized
expertise in the study areas of concern.

Shore-based  camps will  be established  in PWS to  conduct photo-
identification  studies  on killer whales  from  small boats  (May
through September 1991). Camp  locations  will  be  similar to those
set up in 1990.  Camps may be moved during the field season based on
whale distribution data collected during the study.  All camps are
fully  self-contained  with necessary items for  camp and vessel
safety.  Camps will be resupplied with food and essentials at least
twice a month by a vessel chartered specifically for this reason.
Each camp is staffed by at least two biologists and one small boat.
Camp personnel will communicate among themselves via marine radios.
For consistency in  data collection, key  personnel  remain in the
field throughout the 5-month period.

Weather permitting,  field personnel will spend an average of 8 to
10 hours  per day conducting  boat surveys searching  for whales.
Effort must be  comparable to the 1989 and 1990 seasons.   Specific
areas,  known for whale  concentrations,   are  investigated first.
However, if reports of whales are received from other sources (e.g,
sighting network described below),  those areas are examined.   If
whales are not located  in "known"  areas and opportunistic sighting
reports are  not available, a  general search  pattern is  developed
and  implemented.   Travel  routes typically  taken  by whales  are
surveyed.  When whales are sighted, researchers stop further search
efforts and  approach  the whales  to collect  photo-identification
information.   When  whales  are encountered, researchers select a
vessel  course and speed to  approximate the animals7 course and
speed to facilitate optimal photographic positioning.

To obtain a  high-guality  photograph,  an approach within  30-60
meters is required.  Photographs  are taken of the left side of the
whale's dorsal  fin and saddle patch.  Any high-performance camera
system  can  be  used  to  collect  the  data.    Motor   drives   (5
frames/sec)  and 300 mm fixed lenses are optimal.   The camera
shutter speed is set  to  I/1000th  second,  or the  highest  speed
possible.  The film type should allow for a high shutter speed and
good depth of field.  For this project black and white ASA  400 film
is used and developed  at  ASA 1600.   The camera should  be held
steady  and  be  supported by  a shoulder  brace  if possible.   All
exposed  film during this  study  will be  developed by  the same
photographic  laboratory.   Film will  be  processed  throughout the
season to allow field personnel to obtain necessary feedback within
                                11

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two weeks of encounters.  Proper labelling of exposed film includes
date, roll number, photographer's initials, location, species code,
and ASA setting.  A new roll  of  film is used for each encounter.

Daily effort logs are maintained  each  day which will  permit 1)
quantification  of the  amount of  time  searching  for  whales vs
photographing whales,   2)  quantification of search  effort under
different weather conditions, 3)  daily vessel trackline, and 4) an
estimation of number of vessels/aircraft encountered in the study
area.

To  increase the  sighting  effort within  PWS  to ensure that  all
whales are  being  seen  and  photographed,  a marine mammal sighting
network will be organized  throughout the  PWS  area.   This  network
will record  all opportunistic sightings  of whales collected from
Alaskan  State  Ferries  and private  aircraft  and  boats.    Whale
sightings  are  reported directly to  the whale  research vessels.
Field teams  respond  by searching out the area  where  whales were
reported in order to collect photographic data.

All exposed film of killer whales collected during the 1991 field
season  will be  analyzed  for individual  identification.    Each
negative  (or  print  as  needed)  is placed  under  a  dissection
microscope for identification purposes and notes and sketches are
made.   Sub-standard photographs  (not showing  enough  detail or
improper angle/side)  are discarded, thus reducing the probability
of  mis-matching photographs.   Photographs are then grouped by
individuals.  Each identified whale  is  then visually compared to
the historical photographic database available.  Once an individual
whale is properly  identified, it is relatively  easy to identify the
pod to which it belongs.  When all photographs are properly entered
and evaluated, it is then possible to determine 1) if all members
of  the  pod  were  present,  and  2)  if pod  structure/integrity is
similar  to previous years.   Missing animals are noted.    It is
imperative  that  1991   studies  be  done  to  verify  the  missing
individuals described in 1990.  The stability of  resident pods over
time is  such that if  an  individual  is  listed  as missing  for at
least one year, that missing whale is considered dead.

To avoid biases in data interpretation,  it is important that the
amount of effort in searching for and photographing whales in  1991
is at least equal  to  (but not less than) that completed in previous
years.  For a large pod (>12  animals), the likelihood of obtaining
photographs  of all  individuals is  increased  as  the  number of
encounters is increased.  Some individuals, and certain pods, are
more likely to approach vessels,  making photographic documentation
easier,  while others remain considerably distant, making for more
difficult  conditions.    Whale behavior  also  plays  a  role when
attempting  to  obtain photographs of individuals.  If  the  pod is
resting  (typically  grouped  together),   it is  easier  to  obtain
photographs of all whales versus when the pod is  travelling  (spread
out  through  an  area).    Researchers  with   prior  killer  whale

                                12

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experience in  a  particular area, who are  capable of recognizing
individuals,  will also enhance the likelihood of accounting for all
whales within a pod.

Calves of  the  year will  be noted and their  mothers identified.
Natality (number of calves per adult female) will be calculated for
each pod for each  year  and comparisons made between resident and
transient  groups  using  descriptive statistics.   Mortality rates
through  1990  will  also  be  calculated  for  resident  groups.
Mortality for transient  pods will be calculated when necessary data
are available.

General location of whales will be recorded each time photographs
are taken, allowing comparisons of pod distributions among years.
Changes in normal distribution patterns will be reported.
                           BIBLIOGRAPHY

The following killer whale articles  are pertinent to the studies
being conducted in Alaska.

Anon. 1982.   Report on the workshop  on identity,  structure, and
     vital rates  of killer whale populations.   Rept.  Int.  Whal.
     Commn, 32: 617-631.

Balcomb, K. C.  1978.  Orca Survey 1977.  Final report of a field
     photographic  study  conducted  by   the  Moclips  Cetological
     Society  in  collaboration  with  the U.  S.  National  Marine
     Fisheries  Service  on killer whales  (Orcinus  orca)  in Puget
     Sound.  Unpub.  Report to  the Marine Mammal Division, National
     Marine Fisheries Service, Seattle,  Washington, 10 pages.

Bigg, M. A.   1982.   An  assessment  of killer whale (Orcinus orca)
     stocks off Vancouver Island,  British Columbia.   Rept.  Int.
     Whal. Commn., 32: 655-666.

Braham,  H. W. and M. E. Dahlheim.  1982.  Killer whales in Alaska
     documented in  the  Platforms of  Opportunity  Program.   Rept.
     Int. Whal. Commn. 32: 643-646.

Calambokidis, J. , J. Peard, G.  H.  Steiger,  J.  C.  Cubbage,  and R.
     L.  DeLong.   1984.   Chemical  contaminants  in marine mammals
     from  Washington State.   Natl.   Oceanic  Atmospheric Admin.,
     Tech. Memo, NOS QMS,  6: 1-167.

Ellis,  G.    1987.   Killer whales  of  Prince  William  Sound and
     Southeast    Alaska.        A   catalogue    of   individuals
     photoidentified,    1976-1986.        Sea   World    Research
     Institute/Hubbs Marine Research Center, Technical Report No.
     87-200.   April 1987.
                                13

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Fowler, C. W.  1984.  Density dependence in cetacean populations.
     In "Reproduction in Whales, Dolphins, and Porpoises".  Eds. W.
     F. Perrin, R.  L.  Brownell,  and D. P. DeMaster.   Rept. Int.
     Whal. Commn., Spec. Issue 6: 373-380.

Hall, J. D.   1981.  Aspects  of the natural history of cetaceans of
     Prince William Sound.   Ph.D.  Dissertation.   University of
     California - Santa Cruz.  148 pp.

Heyning,  J.   E.   and M.  E.  Dahlheim.    1988.    Orcinus  orca.
     Mammalian Species Account,  No.  304, pp.  1-9, 4 figs.

Leatherwood,   S.,  K.  C. Balcomb,  C.  O.  Matkin,  and  G.  Ellis.
     1984.   Killer  whales  (Orcinus orca) of southern  Alaska -
     results  of  field research  1984  preliminary report.   Hubbs
     Sea World Research Institute Tech. Report No. 84-175, 59 pp.

Leatherwood,   S.,  A.  Bowles,  E. Krygier,  J.  D.  Hall,  and  S.
     Ignell.    1985.  Killer whales (Orcinus orca)  in Southeast
     Alaska,  Prince William  Sound, and Shelikof Strait;  A review of
     available information.   Rept. Int.  Whal. Commn., SC/35/SM 7.,
     10 pp.

Perrin, W. F.  and  S.  B. Reilly.   1984.  Reproductive parameters
     of dolphins  and  small  whales of  the family delphinidae.  In
     "Reproduction in Whales, Dolphins,  and Porpoises".  Eds.  W. F.
     Perrin,  R. L. Brownell, and D.  P.  DeMaster.  Rept.  Int. Whal.
     Commn.,  Spec. Issue 6:  97-134.

von  Ziegesar,  0., G. Ellis, C.  Matkin, and  B.  Goodwin.   1986.
     Repeated  sightings of  identifiable killer  whales  (Orcinus
     orca) in Prince William Sound, Alaska  1977-1983.   Cetus, Vol.
     6, No.  2, 5 pp.


                             BUDGET

Salaries                     $ 48.0
Travel                          8.0
Contracts                     110.0
Supplies                       10.0
Equipment                      10.0

Total                         186.0
                                14

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MARINE MAMMAL STUDY NUMBER 5

Study Title:   Assessment of  Injury  to Harbor Seals  in  PWS,  GOA
               and Adjacent Areas

Lead Agency:   ADF&G

Cooperating Agency:   NOAA


                          INTRODUCTION

Harbor  seals  (Phoca  vitulina  richardsi)  are  one  of the  most
abundant  species  of marine  mammals  in PWS.   They  are  resident
throughout the year, occurring primarily in the coastal zone where
they feed and haul out to rest,  bear  and care for their young, and
molt  (Hoover 1988) .   Some  of the largest  haulouts  in  PWS,  and
waters  adjacent to  these  haulouts,  were  directly  impacted  by
substantial  amounts of oil  during the  EVOS.   Oil that moved into
the  GOA  impacted  harbor seal  habitat at  least as  far  to  the
southwest as Tugidak  Island.   The impacts of the  EVOS on harbor
seals are of particular concern since trend surveys indicate that
the  number  of  harbor seals  in  PWS declined by 40%  from  1984  to
1988, and similar  declines  have  been noted  in other parts of the
northern GOA (Pitcher 1989).

During the EVOS, harbor seals were exposed to oil  both in the water
and on land. In the early weeks of the  spill they swam through oil
and inhaled aromatic hydrocarbons as they breathed at the air/water
interface.  On haulouts in oiled areas, seals crawled through and
rested on oiled rocks and algae throughout the spring and summer.
Pups were born on  haulouts in May and June, when some  of the sites
still had oil  on  them, resulting in pups becoming  oiled.   Also,
many pups nursed  on oiled  mothers.   At haulouts  throughout the
oiled areas, seals  were exposed to greatly increased human activity
in the form of air and boat traffic and cleanup activities.

Following the EVOS, field observations  were made of seals in oiled
and unoiled  areas  of PWS.   Carcasses of 47  seals were necropsied
and sampled; 19 were found dead or died in captivity, and 28 were
collected specifically for sampling.  Preliminary histopathological
and toxicological analyses are almost complete.

In 1989  and 1990,  aerial surveys  were conducted during  June  to
count the number of harbor seal pups and non-pups on  25 oiled and
unoiled haulouts  in PWS.   Results from the two years have been
compared  to determine  whether  the  number  of pups/non-pups  is
similar  in  oiled  and  unoiled areas and  whether  the proportion
changed from 1989 to 1990.  Aerial surveys were also  conducted at
the same 25 haulouts during the fall  molt.   Results  of fall 1989
                                15

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and 1990 surveys have been compared to results of surveys flown in
1984 and 1988 to determine whether trends in numbers are similar in
oiled and unoiled areas.

This   project   proposes   to   complete  histopathological   and
toxicological analyses of harbor seal tissues  and to provide counts
of harbor  seals on  haulouts  in oiled  and unoiled parts  of  PWS
during pupping and molting in 1991.   Data from this third year of
aerial surveys  following the spill will be used to evaluate whether
1990 data were  indicative of a normal year and whether changes that
occurred  in  the  distribution  and  abundance  of  harbor  seals
following the EVOS coincided with the presence or absence of oil in
the area or on  haulouts.   Toxicological analyses  of  tissues from
oiled  seals  will  allow an assessment  of  how hydrocarbons  were
assimilated by seals and how contaminant levels changed with time;
analysis of tissues from control seals will provide baseline data
for comparison with  results from  seals  collected  in  oiled areas.
Final analysis  and interpretation of histopathology  slides  will
provide thorough documentation of  toxic  damage to tissues.  Survey
and laboratory data, in combination with historical data for PWS,
will be used to evaluate whether the EVOS caused a reduction in pup
productivity at oiled  sites,  and whether changes in  abundance
during the fall molt were due to the EVOS.  This information will
be used to  make recommendations  regarding restoration of lost use,
populations, or habitat where injury is identified.


                           OBJECTIVES

A.   Test the hypothesis that harbor seals found dead in the area
     affected by the EVOS died due to oil toxicity.

B.   Test the hypothesis that seals  exposed  to  oil from the EVOS
     assimilated   hydrocarbons   to  the  extent  that   harmful
     pathological conditions resulted.

C.   Test the  hypothesis  that pup production was  lower  in oiled
     than in unoiled areas, or  than  in  years not  affected by the
     EVOS.

D.   Test the  hypothesis  that the number of harbor  seals on the
     trend count  route during  pupping  and molting  decreased in
     oiled areas of PWS as compared to unoiled areas.
                             METHODS

In 1991, aerial surveys will  be  conducted during pupping in June
and molting in September along a previously established trend count
route (Calkins and Pitcher 1984; Pitcher 1986, 1989)  that covers 25
haulout sites  and includes  6 sites impacted  by  the EVOS (Agnes,
Little Smith,  Big Smith,  Seal, and Green islands,  and Applegate

                                16

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Rocks), 16 unoiled  sites,  and  3  intermediate sites that were not
physically oiled but were adjacent to oiled areas.  Visual counts
will be made of seals at each site and photographs taken of large
groups for later verification.

During June,  separate  counts will be made  of  pups and non-pups.
Pupping surveys are needed  in  1991  since there are no historical
data available  from PWS during the pupping  season with which to
compare the 1989 results, and a single year of post-spill data from
1990 is not enough  to establish what is normal in a non-oil-spill
year.

Surveys during the molt in  1991 are necessary to determine whether
observed changes in the number of seals on oiled sites between 1988
and 1990 persist.

All  statistical tests  for  significance will  use alpha  = 0.05.
Statistical testing is not appropriate for  all  objectives.   The
assessment of cause of death of animals  found in areas impacted by
the EVOS  (Objective A)  will require  expert evaluation of limited
and varying toxicology and histopathology data sets.

Toxicological results for each seal collected will  be entered into
a  database  along  with  information on  date and  location  of
collection; presence of oil in the area;  degree of  external oiling
of  the seal;  and  age,  sex,  size,  and  reproductive  condition.
Hydrocarbon levels in the tissues will be tabularized.  Differences
between groups will be  tested where possible using ANOVA  (Neter and
Wasserman 1974).

Types of pathology  detected will  be  listed for each specimen and
will be  grouped  into  tables by  sex,  age,  collection  location,
and/or degree of oiling. Incidence of pathology will be expressed
as the percentage of the total  number of  animals in the group that
exhibited a particular type of anomaly.   Incidence of pathology
will be  evaluated  in  light of   toxicological results  for  each
specimen.

Harbor  seal   surveys  must  be  conducted  within  biological  time
windows imposed by the pupping and molting  periods.  While results
of previous harbor seal trend counts  have indicated that it is
desirable to  obtain 7-10 counts  during  a  survey period  (Pitcher
1986, 1989) ,  the actual number of counts is frequently limited by
the number of  days suitable  for flying.   During  pupping, the survey
window cannot be extended to accommodate sample size needs since,
as pups grow and are weaned,  they  become  increasingly difficult to
differentiate  from the air.   Similarly,  during  the molt  it is
necessary to confine surveys to the period when  maximum numbers are
hauled out.

Aerial surveys of harbor seals do  not estimate  the  total number of
seals present since they do not account for seals that are in the

                               17

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water or  seals hauled  out  at locations not  on the  trend count
route.  Surveys provide  indices of abundance based on the number of
hauled out seals counted.   Interpretation  of  trend count surveys
relies on  the assumption that counts  of harbor seals  on select
haulout sites  are  valid linear  indices  of local  abundance.   We
assume that within a given biological window,  such as the pupping
or molting period,  haulout behavior remains the same from one year
to the next,  and counts can thus be compared.   Standardization of
procedures minimizes  the affects of variables  such  as  tide and
weather that could influence the  number  of seals hauled out on a
given day.

The trend count route includes haulputs  impacted by  the EVOS, as
well as haulouts that are  north, east, and south  of  the primary
areas impacted by oil.  There is an adequate sample of both oiled
and unoiled areas.

Data from  1991 pupping surveys  will  be used in a retrospective
analysis comparing counts of non-pup seals in oiled  and unoiled
sites between years  (1989-91)   and  using  the   same  statistical
techniques employed for fall molting surveys (Frost 1990).

In order  to  compare  pup production at  oiled and unoiled sites
(Objective C) , a  one-way  analysis  of co-variance  (Neter  and
Wasserman  1974)  will be performed on the square roots  of the
trimeans  (Hoagliri  et  al. 1985)  of  pup counts,   using the square
roots of non-pup trimean counts as the  covariate.  The square root
transformation will be used to correct for non-constant variation
of the count data  (Snedecor and  Cochran 1980).   Linear contrasts
(Neter and Wasserman  1974), where the average number of pups is
adjusted to  a common number of non-pups,  will  be used  to test
working hypotheses.

Data collected during the molt in 1984, 1988,  1989, and 1990 will
be used for comparisons with data collected in  1991.   A repeated
measures ANOVA (Winer  1971) will  be  performed on  the trimean
(Hoaglin et al. 1985) of the site count data  in order to examine
trends in  abundance  at  oiled versus unoiled  sites.   The trimean
statistic will  be  used  as a measure of  central  tendency because
sets  of  counts at  a  single  location sometimes show  bimodal
distributions or include extreme variations. This analysis assumes
random  samples,   constant  variance,   and  normality   of  the
differences.   If necessary,  transformations (Snedecor and Cochran
1980) will be used  to  ensure constant variance and  normality.  The
test assumes  that the mean proportion of the population hauled out
on the  trend  count  route  is  constant  over  years.   Hypotheses
addressing Objective  D  will be  tested  using orthogonal contrasts
derived from the ANOVA.
                                18

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                           BIBLIOGRAPHY
Calkins, D.,  and K.  Pitcher.
     southern Alaska: 1983-84.
     16pp.
    1984.   Pinniped investigations  in
    Unpubl. Rep.  ADF&G,  Anchorage, AK.
Frost, K. J.  1990.  Marine Mammals Study Number 5: Assessment of
     injury to harbor  seals  in Prince William Sound,  Alaska, and
     adjacent  areas.    State-Federal  Natural  Resource  Damage
     Assessment  for  April 1989-December  1990.    Unpubl.  Prelim.
     Status Rep. ADF&G Fairbanks, AK.  22pp.

Hoaglin, D. C.,  F. Mosteller,  and J.  W.  Tukey.   1985.  Exploring
     data tables, trends,  and shapes.  John Wiley & Sons. New York,
     N.Y. 527 pp.

Hoover, A. A.  1988.   Pacific  harbor seal.   Pages 125-157 in; J.
     W. Lentfer  (ed).  Selected Marine Mammals of Alaska: Species
     Accounts with Research and Management Recommendations.  U. S.
     Marine Mammal Commission, Washington, D. C.

Neter, J.,  and  W.  Wasserman.   1974.   Applied linear statistical
     models.  Irwin,  Inc., Homewood, IL.  842 pp.

Pitcher, K. W.  1986.  Harbor seal trend count surveys in southern
     Alaska, 1984.  Unpubl. Rep. ADF&G, Anchorage, AK. 10pp.

Pitcher, K. W.  1989.  Harbor seal trend count surveys in southern
     Alaska, 1988.  Final Rep.  Contract MM4465852-1 to U.S. Marine
     Mammal Commission, Washington, D.C. 15pp.

Snedecor, G. W.  and  W. G. Cochran.   1980.   Statistical methods.
     Iowa State University Press, Ames, 10.  507 pp.

Winer, B. J.  1971.   Statistical principle in experimental design.
     2nd Ed.  McGraw-Hill, New York, N. Y. 907 pp.
Salaries
Travel
Contracts
Supplies
Equipment

Total
  BUDGET

 $54.6
    8.3
   28.5
    2.8
	Q

 $  94.2
                                19

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MARINE MAMMAL STUDY NUMBER 6A

Study Title:   Boat Surveys  to  Determine Sea Otter  Abundance in
               PWS Following the EVOS

Lead Agency:   FWS


                           INTRODUCTION

In the first year  following  the  EVOS, hundreds  of sea otters are
known to  have died  as a  result of contamination  by oil.   The
capacity of the population to recover to pre-spill  levels is not
known.  This  study will assess  the  impacts of the  oil  spill on
Alaska sea otter populations through surveys  of wild populations
living in oiled and unoiled areas.
                           OBJECTIVES

A.   To  test the  hypothesis that  sea otter  densities are  not
     significantly different between oiled and unoiled areas.

B.   To  test the  hypothesis that  sea otter  densities are  not
     significantly different  between pre- and post-spill  surveys in
     oiled and unoiled areas.

C.   To estimate the magnitude of any change between pre- and post-
     spill sea otter population estimates in PWS.

D.   To estimate post-spill population size of sea otters in PWS.

E.   To estimate winter 1991 offshore  densities  of  sea  otters in
     oiled and unoiled areas to  estimate  otter density  values at
     the time of the oil spill in March 1989.


                             METHODS

An original  boat-based survey of  PWS  consisted of a complete sea
otter  census of 718  shoreline  transects  totalling 4,062  km of
shoreline (Irons et al. 1988).   This initial survey was conducted
using  a  single  vessel  over a period of two field seasons  (June,
July,  and  August  of   1984  and  1985) .    A  random   sample  of
approximately 25 percent  of  the transects  was  surveyed in June,
July,  and August  of  1989.   In addition,  offshore areas were
surveyed in  July and August  1989.   These  same transects, .plus an
additional 25  shoreline transects, were  again  sampled in June,
July, and August of 1990.  A slightly reduced sample of shoreline
and  offshore transects  were surveyed  in  March 1990.    Surveys
proposed  for 1991  include  replication  of the  March   and July
surveys.

                                20

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To insure that project design and standard operating procedures are
followed,  (1) all crew members will read and discuss the observer
guidelines handbook,  (2)  all crew members will  partake in trial
surveys prior to actual surveys, (3) one person on each boat will
have  responsibility for  maintaining  consistent  data  collection
procedures,  (4)  standardized  forms  will  be used  during  data
collection,  and (5) data  forms will be checked by the project
leaders at the end of each day to insure the integrity of the data.

Post-stratification of shoreline and offshore transects by presence
or absence of oil has been based on data collected by the Coastal
Habitat Study,  the  Air/Water  Studies,  and the Technical Services
Study Number 3.

Prior to the start of  each survey, transect and environmental data
are collected and recorded on a standard data sheet.  Transect data
consist of observer  names, transect number, date,  and start time of
transect.    Environmental data  include  air temperature,  water
temperature,  sea state,  wind  direction and velocity,  weather,
presence  of  ice on transect, and  tidal state.    In  addition,  an
overall  observation condition  is  recorded,  and notes  on  human
activity  and presence  of  oil  within  the  transect  are  taken.
Surveys   are  postponed   or   aborted   in  unsuitable  conditions
(visibility less than 100 m, or wave heights greater than 2 ft).

Shoreline  transects from Irons et  al.   (1988) are surveyed  at a
speed of  5-10  knots from  100 m offshore.   Distance to shore is
periodically checked  using a rangefinder.   One  observer surveys
from the  shoreline  to the boat, while  a second  observer surveys
from the  boat  seaward  an additional  100  m.  The survey  window
extends approximately  100 m ahead  of,  and 100 m above the boat
while travelling.   Sightings  of marine  mammals,  birds,  and boats
within this window are recorded on the standard data sheet as being
within the "inside"  strip  (0-100 m)  or  the "outside" strip (100-200
m) .  In addition  to species,  strip, and quantity, information is
collected  on the  disposition of the sighting  (object was  in the
water, in the air, on land, or following the boat).  Deviation from
the transect due to rocks, ice,  or other obstructions is noted in
the comments section of the data sheet.

Offshore  transect lines are  oriented  along north/south axes, and
steered   by   a   combination  of  compass   heading   and  LORAN-C
interpolator.  Boat speed for offshore surveys is slightly faster
than for  shoreline  surveys,  ranging from  15-25  knots,  dependent
upon sighting  conditions.   Transect  and environmental  data are
collected  as  in shoreline  surveys.     The  sampling  window  is
essentially the same as well, with observers sampling a strip
100 m in width on each  side of the boat,  and  forward approximately
100 m.   By definition,  shoreline surveys sample  the 200 m strip
adjacent  to shore.  For the  purposes  of this study,  the offshore
environment is  therefore  defined  as any area greater  than  200 m
from shore.   Objects  further than 200 m from shore  are recorded

                                21

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within the  "offshore"  strip on the  data sheet.   Where offshore
transect lines  intersect land, objects  sighted within 200  m of
shore are recorded within the "nearshore" strip.
                          DATA ANALYSIS

Statistical  assumptions  pertinent  to these  analyses have  been
outlined in the previous study plans.  Data collected during 1989
and 1990 suggest that these assumptions are being met.

Abundance estimates will be calculated independently for shoreline,
coastal and pelagic environments using ratio estimator techniques
according to Cochran  (1977).  Estimates calculated from third-year
surveys will be compared to earlier estimates for the determination
of injury to the  sea  otter population within  PWS.  Differences in
otter densities  will  be tested  using two  sample t-tests and/or
ANOVA, dependent upon post-stratification of oil condition.
                           BIBLIOGRAPHY

Cochran, W.G. 1977.  Sampling techniques. John Wiley and Sons, Inc.
     New York, New York. 428pp.

Irons, D.B., D.R.  Nysewander,  and J.L. Trapp. 1988. Prince William
     Sound sea otter distribution in relation to population growth
     and habitat  type.  U.S.  Fish  and Wildlife  Service.  Unpubl.
     report. 3Ipp.


                              BUDGET

The costs of this study are included in the budget for Bird Study
Number 2 and totals  $220.0.   The budget  breakout is not repeated
here.
                                22

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MARINE MAMMAL STUDY NUMBER 6B

Study Title:   Intersection Model of Sea Otter Mortality

Lead Agency:   FWS


                           INTRODUCTION

Following the release and subsequent movement of oil from the EVOS,
live and dead oiled sea otters were observed within PWS and along
the KP.  Oiled sea  otter  carcasses  were retrieved and live oiled
otters were  captured for transport to  rehabilitation centers in
Valdez,  Seward,  and  Homer.    The  number  of  dead oiled  otters
retrieved may include some otters that were dead before the spill.
It is likely that additional otters became oiled and died and their
carcasses were not  recovered, while others  may have  become oiled
and survived.

Three approaches are currently under investigation to  estimate the
number of sea otter mortalities that resulted from acute exposure
to oil.  One method estimates the number of unrecovered carcasses
based  on the probability  of carcass recovery.   Another  method
compares estimates  of sea otter  abundance  before and  after the
spill.  The third approach uses an analytical model to relate oil
exposure to  subsequent  mortality of sea otters.  The purpose of
this study is to develop such a model for application along the KP.
This model  may  be  extended for  application  throughout  the spill
zone to provide an estimate of the total acute mortality.

This  approach  involves:     1)   estimating  the  abundance  and
distribution  of  sea  otters  in  near-shore and  off-shore  habitat
along the KP at the time of the spill,  2) estimating the level of
exposure of each otter, 3) estimating the degree of oiling received
by otters at each exposure level, and 4) estimating the mortality
rate associated with each degree of oiling.   Sea otter oiling and
population  data  along  with  the  oil distribution  data will be
integrated by the model to provide an estimate of the total spill
induced mortality for this area.


                           OBJECTIVES

To develop  an analytical  model  capable  of estimating rates of
exposure of  sea  otters to oil,  degree  of  oiling, and mortality
along the KP following the EVOS.
                                23

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                             METHODS

Oil Distribution

A hind-cast computer model developed by NOAA (on-scene spill model,
OSSM) will be used to simulate the distribution of oil particles as
they traveled through PWS and along the KP.  The  OSSM model traces
the  movement  of  10,000 particles  (Lagrangian elements,  each
representing about 1,100 gallons of oil) from their origin at Bligh
reef, at three hour intervals.  Under this model,  about 1,250 (12%)
of the oil particles moved out of PWS and along the KP.

Sea Otter Abundance

The abundance and distribution of sea otters in near-shore and off-
shore habitat along the KP at the time the oil passed through will
be estimated based on  the spring  1989 helicopter survey that was
conducted during the spill response.  The location of each observed
otter was recorded during the survey on  large scale maps.   These
locations and numbers of otters will be used as an estimate of the
distribution and abundance of otters at the time of the spill.

Exposure to Oil

In order to measure exposure, an  exposure region will be defined
for each otter or  group of otters, as a circle with radius 1.4 km
centered at the otter's location during the survey. Any portion of
this circle that overlaps land will be excluded from the exposure
region.   The 1.4  km radius  represents the average  distance sea
otters were observed to move between successive radio relocations
recorded between 18 and 36 hours apart in California (Rails et al.
1988).  The Rails et al.  (1988) data include movements of adult and
sub-adult male and female sea otters.

The number of gallons per day times the number of  days that oil was
within an  exposure region divided by the  area  of  the exposure
region (gallon*days/km2)  will be used as a measure of the exposure
of that location to oil.  The proportion of  the observed otters at
each  location will be used  to  estimate  the proportion  of  the
population with that location's level of exposure.

Study Areas

Data for relating exposure levels to oiling and mortality of otters
were collected within two areas of PWS.  The first of these areas
was Herring  Bay on the north end  of Knight  Island  where heavy
oiling was observed to persist over time,  all otters were oiled,
the degree of oiling was heavy and  mortality rates were high.  The
second  site comprised  the  northeast  third  of  Prince of Wales
Passage, including Iktua Bay  between Evans and Bainbridge Islands.
This area was  lightly  oiled  along most of  the shoreline  and oil


                               24

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appeared to  pass through in  a  short time. Most  sea  otters were
either non-oiled or lightly oiled and mortality was relatively low.

Mortality rate calculations exclude pups born  in captivity, otters
with an undetermined  oiling  status and otters exhibiting obvious
non-oil related  pathology  (eg.,  paralysis  or  blindness).   Bodkin
and Weltz (1990) describe a pattern of declining degree of oiling
and resultant mortality as the time interval between exposure and
capture increased  in PWS.    This pattern  led to  diminishing sea
otter capture efforts in PWS on April 21, 1989, and a shift in the
effort to the KP where initial oiling occurred on or about April 1,
1989.

Sea Otter Capture

During the first 3 weeks of April  1989, otters in Herring Bay and
Prince of Wales  Pass  were  captured with dip-nets  and tangle-nets
(Bodkin and  Weltz  1990).   Each otter was  classified  into  1 of 4
categories based on the quantity of oil observed on its pelage at
the time of capture. The degree of oiling  categories were defined
as follows:  heavy =  complete or nearly complete  coverage of the
pelage with visible oil,  moderate = partial oiling of about 25-50%
of the pelage with visible oil, light = oil not easily visible or
detectable, or a small proportion  (<10%) of the pelage containing
visible oil, and none = oil not visually or tactically evident on
the pelage.

Relating Mortality to Degree of Oiling

Oiled otters were transported to rehabilitation centers, where they
were cleaned and held.   Mortality rates  for each of  the oiling
categories following capture and holding were  recorded.  Mortality
was  considered  spill related  if  it  occurred within 30  days  of
capture. Mortality usually occurred within 5 (65%) days of arrival
(mean =7.1 days; range 0 to 34 days) at a rehabilitation center.

Mortality rates used in the model are based on the mortality rates
observed  in  the  rehabilitation  centers  and  on  a  study  of
experimentally oiled captive sea otters  (Kooyman and Costa 1979).

Relating Degree of Oiling to Exposure

The relationship between the degree of oiling and the exposure will
be estimated by calculating the exposure in gallon*days using the
OSSM and relating that to oiling which occurred in  the study areas.
Values defining high exposure,  moderate exposure,  and low exposure
will be defined for each area.

The proportion of the estimated total near-shore and off-shore KP
sea otter population in high, moderate, and  low exposure categories
will be determined  based on their estimated exposure values and the
scale developed for the study areas.  The  total mortality will be

                                25

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estimated by taking each  of  the  products  of the total population
estimate, the exposure level  proportion, a corresponding degree of
oiling  proportion and  its  associated mortality  rate,  and then
summing over the  degree of oiling  categories.   Overall mortality
rates for each  exposure category will be estimated  based on the
mortality rate and the size of each portion of the population.  The
total mortality for the KP otters will be estimated as the sum of
the totals for the three exposure categories  for  the near-shore and
off-shore habitats.
                          DATA ANALYSIS

A  point estimate  of sea  otter mortality  resulting  from  acute
exposure to oil along the KP will be obtained as described in the
Methods section.
                           BIBLIOGRAPHY

Bodkin, J.L. and F. Weltz. 1990.  A summary and evaluation of sea
     otter capture operations in response to the Exxon Valdez oil
     spill,  Prince William  Sound  Alaska.   In  K.  Bayha  and J.
     Kormendy,  eds.,   Proceedings  of  the  Sea  Otter  Symposium,
     Anchorage, Alaska. April 17-19, 1990.  In press.

Kooyman,  G.L.  and D.P.   Costa.  1979.    Effects  of  oiling on
     temperature   regulation  in  sea   otters.     Report,  Outer
     Continental Shelf Environmental Assessment Program, N.O.A.A.
     Contract No.  03-7-022-35130.  25pp.

Rails, K., T. Eagle, and D.B. Siniff.  1988.  Movement patterns and
     spatial use of California  sea  otters.  In D.B.  Siniff and K.
     Rails, eds., Population status of California sea otters.   U.S.
     Fish  and Wildlife  Service,    Minerals  Management  Service,
     Contract No.  14-12-001-30033.   368pp.


                              BUDGET

Salaries                      $70.0

Total                         $70.0
                                26

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MARINE MAMMAL STUDY NUMBER 6C

Study Title:   Radiotelemetry Studies on Sea Otters in PWS

Lead Agency:   FWS

                           INTRODUCTION

On  March  24,  1989,  over  11  million gallons  of crude  oil  were
spilled in  PWS due to  the EVOS.   Thousands of sea  otters  were
potentially  affected.   Exposure of  sea otters  to  components of
crude oil may have caused acute illness and mortality  or chronic
illness which may  cause  population  damage  either due to eventual
mortality, reduced production or both.

Within months  of  the  spill, research was  initiated to determine
both the acute and the  chronic  consequences of exposure to crude
oil from the EVOS  on sea otters  that  were not treated and remained
in the affected habitat,  as well as  on otters that were treated at
otter rehabilitation  centers  following exposure.   From the  wild
population,  100 adult and  64  dependent  sea otters were captured,
examined,  instrumented with radio-transmitters,  and monitored in
PWS beginning in October  1989 to the present. Additionally, of the
large number of sea otters that were captured  and brought  into
otter rehabilitation  centers,  45 were  radio-instrumented during
June 1989, released in  eastern  PWS  during  July,  and continuously
monitored until the present.   The goal of this research effort was
to provide data on the survival,  reproduction, and behavior of the
sea otters following release from these centers,  and by doing so,
to  gain insights  into both the damage done  to the  PWS sea otter
population and in the efficacy of the "rehabilitation" strategy.

The  studies  proposed herein  represent  a  continuation of  the
research  effort briefly  described  above.    These  studies  were
designed to  permit comparisons  of  certain  characteristics of the
sea otters in the  oil  spill zone not  only with those of sea otters
from  eastern  PWS, but  also  to  information about sea  otters
throughout PWS available from previous studies dating back to the
mid-1970's.  This  approach provides  both a coincident baseline for
the data  gathered  on  sea otters in  the spill zone  and  a way to
address the  question  whether  the   spill  may have directly  or
indirectly caused  damage over a larger geographic  area than has
usually been assumed.   Additionally, it provides a way to gauge
what is normal for this  population,  and in so doing, establishes
both a measure and a goal for recovery efforts.

In addition to the general goals described above, the information
gathered during these studies will provide information crucial to
formulating  restoration policy  for  sea  otters throughout the oil
spill zone,  including  information on habitat utilization, and more
specifically, identification of  critical habitats, recolonization
                                27

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rates, predicting and monitoring population growth rates during the
recovery  phase,  and   the  formulation  of  future  response  and
restoration policies for sea otters throughout their range.


                            OBJECTIVES

Weanlings

A.   To test the hypothesis that weanling survival at various age
     intervals is not different between oiled and unoiled areas.

B.   To document the movements  of weanling sea otters with respect
     to areas in PWS that have been affected by the EVOS.

Adult Females

A.   To test the  hypothesis that pup survival  pre-weaning is not
     different between oiled and unoiled areas.

B.   To test  the hypothesis  that  survival  of adult female sea
     otters is not different in oiled and unoiled areas.

C.   To test the hypothesis that pupping rates of adult female sea
     otters are not different between oiled and unoiled areas.

D.   To document  the  movements of  adult  female sea  otters with
     respect to areas  in PWS  that  have been affected by the oil
     spill.

Otters from Rehabilitation Centers

A.   To test  the  hypothesis  that  survival of  sea  otters that
     underwent  oiling,  cleaning,  treatment,  and release  is not
     different from that  of sea otters that were not affected by
     the EVOS.

B.   To test the hypothesis that reproductive rates of female sea
     otters that underwent oiling,  cleaning and treatment does not
     differ significantly from that of female sea otters that were
     not affected by the EVOS.

C.   To document the movements of sea otters from treatment centers
     relative to impacted habitats  in western PWS and the KP.
                             METHODS

No additional capture or examination of  sea otters is proposed for
this study.  Capture, instrumentation, and biological sampling of
study otters has  been well described in the  1989  and 1990 study
plans.

                                28

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Radio-instrumented  sea  otters will be monitored  by observers in
aircraft and  skiffs.   Aircraft and skiffs will  be equipped with
right-and left-mounted Yagi antennas and programmable, scanning FM
receivers.  Aircraft  will  be flown at variable heights depending
upon whether  observers  are attempting to  locate radio signals or
make visual observations on individual sea otters.   An attempt will
be made to find and visually examine each otter at least biweekly
until 30  September, 1991.    After  that date, we  will  locate and
determine  the  status  of  each  otter once  per  month until  15
February,  1992.    Data will  be  recorded  directly  on  xeroxed
topographical maps  and  on  data sheets for later  data  entry into
computers.

Information on  presence or absence of oil  will  come  from data
collected in the Coastal Habitat Study, Subtidal Studies, Technical
Services Study Number 3, and response data sets.
                          DATA ANALYSIS

A.  Tests

It is assumed that control animals, from unoiled portions of PWS,
are healthy and relatively uncontaminated,  and that their survival
is representative of that of wild populations.  It is also assumed
that  sea  otters captured  in  the treated  areas have  been either
directly or indirectly exposed to the spilled oil.

B.  Analytical Methods

Survival analyses will be conducted using the Kaplan-Meier product
limit estimator  (Kaplan  and Meier  1958, White and  Garrott 1990)
programmed  in  a simple  Lotus 123  spreadsheet and  plotted using
Lotus Freelance  graphics software.   Significance  of differences
between control and treatment groups will be tested following the
procedure described by Cox and Oakes  (1984).

Reproductive data will be compared between treatment and control
groups using contingency  tables and tests of independence.  Two-way
contingency tables will be used except when interactions among age,
sex, treatment type  or  location are of interest.   In that case,
three-way  or multi-way  contingency  tables  based  on log-linear
models will be used  (Sokal and Rohlf 1981).
                           BIBLIOGRAPHY

Cox, D. R. and D. Oakes.  1984.  Analysis of survival data.
     Chapman & Hall, New York.  201pp.

Kaplan, E. L. and P. Meier.  1958.  Nonparametric estimation from
     incomplete observations.  J. Am. Stat. Assoc.  53:457-481.

                                29

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Sokal, R. R. and F. J. Rohlf.  Biometry.  Second Edition.  W. H.
     Freeman & Co., San Francisco, CA.  859pp.

White, G. C. and R. A. Garrott.  1990.  Analysis  of wildlife radio-
     tracking data.  Academic Press.  New York.  383pp.


                              BUDGET

Salaries                    $  146.0
Travel                          14.0
Contractual                    149.0
Commodities                 	41. 0

Total                       $  350.0
                                30

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MARINE MAMMAL STUDY NUMBER 6D

Study Title:   Sea otter  prey selection and  foraging success in
               Western PWS

Lead Agency:   FWS


                           INTRODUCTION

Sea  otters  commonly  prey   on   a  variety  of  benthic  marine
invertebrates that inhabit coastal waters ranging in depth from the
intertidal to approximately  20 fathoms (Kenyon 1969).  Principal
prey species identified in PWS in the past  include crab, clam, and
mussel  (Calkins  1978; Garshelis  1986;  Johnson 1987).   Damages to
the  nearshore  benthic  community  resulting  from  the  EVOS  may
influence the recovery of affected sea otter populations.  Probable
mechanisms of influence include (1) decreased food availability and
(2) consumption of prey contaminated by hydrocarbons.

Sea otters require a relatively high amount of energy to maintain
their body  temperature  in cold  North Pacific waters  (Costa and
Kooyman 1984) .  Juvenile and  adult sea  otters  consume between 20-
30% of their body weight  per day (Kenyon 1969).   In western PWS,
sea otters spend approximately 50% of a 24  hr period foraging, and
during  the  winter  months   (November-April)  foraging  activity
increases (Garshelis 1986).

To evaluate hydrocarbon contamination in PWS, certain shellfish and
coastal sediments have been systematically sampled  in portions of
the  sea  otter range  by the  Coastal Habitat  and   Fish/Shellfish
damage assessment studies.   Additional taxa  of  shellfish  of sea
otter prey will be collected as needed.

There are at least two functional responses to a contaminated prey
base.   Prey selection  may  continue as  prior  to   contamination,
resulting  in ingestion   of  hydrocarbons  by  sea   otters.    The
consumption of contaminated prey may increase the metabolic demands
on the sea otters' energy budget,  which in turn may retard recovery
of  the  population.   Alternatively,  sea  otters  may reduce  or
eliminate, through prey  selection,  contaminated prey from their
diet.  If sea otter populations  are  limited by food resources in
PWS, as suggested by Johnson (1987), a decline in abundance or a
lack of recovery  of  the  sea otter population may  result.   These
injuries may occur over a  time scale longer than previous damage
assessment studies considered.

The purpose of this study  is to  describe the species  composition
and relative frequency of occurrence of prey selected by sea otters
in three locations in western PWS, following the EVOS.  The results
of this study will  quantify  the  extent  to which  sea otters are
foraging on contaminated prey in these areas and allow evaluation

                               31

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of the need for the collection of additional sea otter prey for
hydrocarbon analysis.  Additionally, this  study  may provide data
necessary to quantify the site specific exposure rate of sea otters
to dietary hydrocarbons.


                           OBJECTIVES

A.   To describe prey species  and the relative frequency that each
     prey species is consumed  by sea otters in 3 areas affected by
     the EVOS.

B.   To collect tissue samples of key sea otter prey (indicated by
     frequency of occurrence > .10)  for toxicological analysis if
     not currently  sampled by coastal  habitat  and fish\shellfish
     studies.

C.   To determine  foraging  success  rates in each of  three study
     areas.

D.   To compare prey species  and foraging  success rates from the
     Green Island area to historic data from the same region.

E.   To estimate mean size and determine approximate caloric value
     per prey item.
                             METHODS

Sea otter prey  will be determined at three  sites  within western
PWS.  Study  sites will be near Green Island, Herring Bay, and Drier
Bay (the latter two on Knight Island).   Study sites were selected
based on  several  criteria:   (1)  the location of  intertidal and
subtidal  sampling  sites  for  sediments  and  tissues,   (2)  the
locations from  which  sea  otter  tissue  samples  were  collected
following the spill, (3)  the capture  location of radio telemetered
sea otters,  and (4) the relative degree of oiling at each site as
quantitatively evaluated by an oil exposure model developed by the
NOAA  (OSSOM) .   In  general, the  Herring  Bay site  exhibited the
heaviest degree and persistence of oiling, the  Green Island area
had a patchy distribution of heavy shoreline oiling, and the Drier
Bay site exhibited  intermediate oiling.   The benthic contours of
the Knight  Island  sites are  similar to one another;   the Green
Island area has a more extensive shallow water area.  Hydrocarbon
contamination is assumed to be relatively  uniform within the study
sites, and  levels  of hydrocarbons observed  in sampled prey to be
representative of those prey species being consumed by sea otters.

The  primary method of  data  collection  will be  observational.
Observations will be made with  the aid of high resolution Questar
telescopes and 10X  binoculars.   Data recorded will include sex, age
class of  focal animal  (adult  or  juvenile) ,  number of  prey and

                                32

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relative prey size (A: < 3 cm,  B:  > 3  to  <  6 cm  , C: > 6 cm to < 9
cm, D: >  9  to < 12 cm and E:  > 12  cm) ,  dive  time,  surface time,
success rate and prey item to lowest taxon.  Repeated dives will be
recorded for a focal  animal until a maximum  of  50 identifiable prey
items are observed per  individual  or  until the  animal is lost or
discontinues  foraging.    Radio-implanted sea  otters  from  damage
assessment  studies will be used as focal  animals  when feasible.
Focal animal selection, when more than one otter is feeding at an
observation  site,  will  be random.     A  minimum  sample  of  500
identifiable  prey  items  will  be  recorded at each of  the three
selected geographic areas.  An attempt will be made to distribute
foraging  observations  from all vantage points  within each study
area.  Compiled foraging  data will be compared to species sampled
by the Coastal Habitat and Fish\Shellfish studies.  If an observed
prey species constitutes  more than 10% overall of the sea otter's
prey at any  site and has not  been sampled  in  supporting studies,
samples will be collected.  Sea otter prey will be collected from
forage areas with the aid of SCUBA, and hydrocarbon levels of the
collected prey will be determined by standard analytical laboratory
procedures.    Sampling  protocols  for  identified prey will  be
determined  as  necessary, depending on species, but  will  follow
accepted methodologies.

Data  from  radio-marked  animals   which  are  of  known  age  and
reproductive status will be collected  as  a  priority.  However, the
majority  of observations  will likely be  collected  on unmarked
animals.   Marked and  unmarked  animals  will  be  distinguished in the
data set.  Adult animals  will be categorized as male,  independent
female, or  female  with  a pup.   Juveniles  will be  identified as
small dark-headed otters estimated to be  less  than 24  months of
age.  Dependent otters will be classified as such.

Data will be collected only during daylight hours, during as many
tidal cycles  as  possible.  Tidal state will  be recorded for all
observations.

Information regarding the species,  their density (when available),
number of species, sample location, and  results of toxicological
analysis of tissue  of marine invertebrates  identified  as sea otter
prey species within the foraging study sites will be required from
the Coastal Habitat and Fish\Shellfish damage assessment studies.
                          DATA ANALYSIS

Initial  analysis will  consist of  listing prey  by  species  and
determining the  frequency of  occurrence  for  each prey  type,  by
site.  Mean success rates, dive times and surface intervals will be
estimated by site and prey type.  Differences between  sites will be
tested with ANOVA.   Prey  selection and foraging  success  can be
compared to historic data collected  at Green Island (Johnson 1987)
as comparable techniques will  be used to gather data.   ANOVA or

                                33

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chi-square contingency table analyses, as appropriate, will be used
to  detect differences  among  dive  times,  success  rates,  mean
Kcal/unit effort, relative frequency of  prey  items,  among areas,
and/or between times.  A significance level of  .05 will be used for
all tests.
                          BIBLIOGRAPHY

Calkins, D.  G.  1978. Feeding behavior and major prey species of the
     sea otter, Enhydra lutris,  in Montague strait, Prince William
     Sound,  Alaska. Fish. Bull. 76(1):125-131.

Costa,  D. P.  and G.  L.  Kooyman.  1984.  Contribution  of specific
     dynamic action to heat balance and thermoregulation in the sea
     otter,  Enhydra lutris.  Physiol. Zool. 57(2):199-203.

Garshelis, D.  L. ,  J.  A. Garshelis  and  A. T. Kimker.   1986. Sea
     otter  time budgets and  prey relationships  in  Alaska.   J.
     Wildlife Manage.  50(4):637-647.

Johnson, A.  M.  1987.  Sea otters of  Prince William Sound, Alaska.
     Unpublished Report, U.S.  Fish  and  Wildlife  Service,  Alaska
     Fish and Wildlife Research Center, Anchorage, AK.

Kenyon, K.  W.  1969. The sea otter  in the eastern Pacific Ocean.
     North Amer. Fauna 68.  352 pp.
                              BUDGET

Salaries                      $48.0
Travel                          5.3
Commodities                     8.0
Equipment                       8.9

Total                         $70.2
                                34

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MARINE MAMMAL STUDY 6E

Study Title:   Sea  Otter Mortality  in PWS  Following  the  Exxon
               Valdez Oil Spill

Lead Agency:   FWS


                           INTRODUCTION

Much of the initial work to assess damages to sea otters caused by
the  EVOS  focused  on  sea  otter  mortality as  a result  of  acute
exposure to oil following the spill.  Additional studies have been
directed at identifying possible longterm effects due to acute or
chronic exposure to hydrocarbons  in  the environment.   Systematic
surveys for  beach cast  marine mammals and  sea birds  have been
identified as valuable  for describing  patterns of mortality over
time   (Bodkin   and  Jameson,   in  press).     Changes   in  the
characteristics of mortality (i.e.,  carcass  recovery  rates, age-
class and sex composition of  dying animals) from pre- to post-spill
time periods may be indicative of  groups of animals compromised by
exposure to oil or hydrocarbon residues in the environment.

Kenyon (1969) and Johnson  (1987) documented patterns of mortality
for  sea   otter  populations  within areas  at  various  stages  of
reoccupation.  Findings indicate extremely low  levels of mortality
for  prime age otters  in habitat  recently  occupied.    Levels  of
mortality  for  young of  the year  and  old animals  increase with
length of occupation of  an area.  Recovery rates of prime age beach
cast carcasses  remains  low,  regardless of  length  of occupancy.
These  studies  are  based  on  information gained from  carcasses
collected  on beaches,   and while  they do not  provide  mortality
rates,  they  do  provide  an  age  class distribution  and  carcass
recovery  rates  that can  be  used  to evaluate  annual  changes and
regional differences in mortality characteristics.

The Green Island area,  in southwestern  PWS, has a long established
sea  otter population  and  is within the  oil  spill zone.   Green
Island was the site of  much of Johnson's  work, which provided 10
years of  baseline mortality data for that area,  as well as 10 years
of mortality data  for the more recently established northeastern
portion (Port Gravina) of the Sound which was not directly affected
by oil.

One year  of post-spill data has already been collected on mortality
patterns  in oiled  areas  (Green,  Knight, Naked  and Perry islands)
and a control area  in the eastern Sound (Port Gravina).  Continuing
the beach surveys will provide additional information on post-spill
characteristics of mortality and  the  persistence of changes that
may  be  occurring  relative  to  pre-spill  mortality  patterns.
Additionally, fresh carcasses  will be  collected for necropsy and
samples taken for histopathology and toxicology studies.

                                35

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                            OBJECTIVES

The overall objective of this study is to conduct beach surveys in
three areas of PWS and collect sea otter  carcasses to determine (1)
if mortality patterns (age class and sex distributions, and rates
of carcass deposition) are similar to previous years, and  (2) post-
spill trends  in mortality.   Specific hypotheses to be tested for
each area are:

A.   The proportion  of  prime age carcasses found  in  1991  is not
     significantly different  from proportions found  in  previous
     beach surveys in PWS.

B.   The  proportion  of  female carcasses  found  in  1991  is  not
     significantly different  from proportions found  in  previous
     beach surveys in PWS.

C.   Post-spill levels of carcass deposition  (number of carcasses
     per linear kilometer of beach surveyed) are not significantly
     different from pre-spill levels of mortality in PWS.


                             METHODS

Beaches will be surveyed in three areas:   1)  Green Island in south-
western PWS,  2) Knight  and Naked Islands in  western  PWS,  and 3)
Port Gravina  in northeastern  PWS.   These  beaches  include those
surveyed pre-spill by Johnson  (1987). Control beaches will include
those in the  Hell's  Hole, Olsen  Bay  area  of Port  Gravina.   These
beaches will  be walked  once in the  spring,  after  the snow melts
from the  supratidal  zone but  before summer revegetation occurs,
which may hide old carcasses washed high on the beach.

Skulls will be taken  from carcasses and  a tooth extracted for aging
(Garshelis 1984).  Fresh carcasses will be collected and necropsied
as  soon  as  possible.    Tissue   samples  will be  collected  for
toxicology  and histopathology.   Badly  decomposed carcasses  or
partial remains may  have no  evidence  indicating  the  sex  of the
individual.  In these cases, if a canine is present  and the carcass
is that  of an  adult,  sex may be determined by  canine  diameter
(Lensink 1962, Johnson 1987).

All teeth  will be sectioned  and prepared  according  to  standard
procedures.    Readings   of  age  will be  done by  two  qualified
individuals.   Necropsies will be performed by personnel  at the
University  of  Alaska-Fairbanks,  Institute  of  Arctic  Biology,
whenever feasible.  Samples taken for histopathology will be sent
to the Armed Forces Institute of Pathology.   Tissue  samples will be
taken for toxicological  analysis according to protocols established
by the Analytical Chemistry Working  Group.
                                36

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                          DATA ANALYSIS

Three variables will be analyzed:  1) the proportion of prime age
carcasses, 2) the proportion of female carcasses, and 3) the rate
of carcass deposition (carcasses per kilometer of beach).  Analysis
of each of these three variables will be run separately.

The proportion of prime  age  carcasses will be the most sensitive
indicator of abnormal change  in  mortality.   This variable is not
influenced by many of the confounding variables associated with the
other two, and a significant change in this parameter is the most
meaningful biologically.  Prime age in this study refers to those
age groups with uniformly high survival rates as measured by pre-
spill data,  and,  based  on Johnson  (1987) ,  is  defined  as  those
animals between 2 and 8 years old for the western PWS and 2 to 10
years old for the eastern PWS.

Changes  in the proportion of female carcasses  recovered  could
reflect changes in the proportions of males and females  in the area
due to immigration/emigration or initially high mortality of one
group  at  the  time  of  the   spill.    Changes  may also  reflect
differential levels of continuing mortality between sexes due to
unequal levels of susceptibility to hydrocarbon toxins or unequal
levels  of exposure  to  toxins  because  of  spacial  segregation.
Proportions (age-classes and sex)  will be tested with a Chi-square
contingency table (Zar,  1984).  Initially, data collected in 1991
will  be  compared to  1990 data  for  each  area.   If  significant
differences are not found,  data from 1990 and 1991 will be combined
and compared to pre-spill data (1974-1984).

The number of carcasses  recovered for a given year is variable and
may be  influenced by a  number  of  variables  (e.g., weather and
current  patterns,  yearly  changes   in  otter  distribution  and
abundance) .   For  example,  from 2  to 34  carcasses  were  found
annually  on  Green  Island area  beaches between 1974   and  1984.
However, examining rates  of carcass deposition may be of some value
for describing  patterns  of mortality over time.   For the Green
Island and Port Gravina areas, a t-test using years as replicates
will be used to compare  rates of carcass  deposition on transects
surveyed in 1990-91  to comparable transects  surveyed in 1974-84.


                           BIBLIOGRAPHY

Bodkin,  J.L.  and  R.J. Jameson.   In  press.    Patterns  of marine
     mammal  and  seabird mortality  as  indicated by  beach-cast
     carcasses along the coast of central California (1980-1986).
     Cdn. Jnl.  Zoology.

Garshelis, D.  L.  1983.    Age estimation of  living  otters.  J.
     Wildlife Manage. 48 (2):456-463.
                                37

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Johnson, A. M.  1987.  Sea  otters  of Prince William Sound, Alaska.
     Unpublished Report,  U.S.  Fish and Wildlife  Service, Alaska
     Fish and Wildlife Research Center, Anchorage, AK.

Kenyon, K. W.  1969.  The sea otter  in  the eastern Pacific Ocean.
     North Amer. Fauna 68. 352 pp.

Lensink, C.  J.  1962.   The history and  status of  sea  otters in
     Alaska.   Ph.D. Thesis, Purdue Univ. 188 pp.

Zar, J. H.  1984.  Biostatistical analysis, 2nd Edition.  Prentice
     Hall,  Inc., Eaglewood Cliffs, N.J.
                              BUDGET

Salaries                      $19.7
Travel                          3.3
Contractual                     5.0
Commodities                     8.8
Equipment                       3.0

Total                         $39.8
                                38

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MARINE MAMMAL STUDY NUMBER 6F

Study Title:   Bioindicators of Damage to Sea Otters From Exposure
               to Oil

Lead Agency:   FWS
                           INTRODUCTION

During  1989  and  1990,  damage assessment  studies on  sea otters
included research on populations living in oiled areas in western
PWS.   Sea  otters in eastern PWS have  served  as  a control group.
Assays  on blood components,  sperm  and  testicular   cells,  and
hydrocarbon levels  in  tissue samples have all been  evaluated as
bioindicators of injury to the sea otters.

Adult  female and juvenile  sea otters with radio  transmitters are
being  monitored  in  PWS as part  of the NRDA  studies.   By summer
1991,  they will  have been monitored  for over a year.   Data are
being  collected  on  survival and  reproductive  rates,  and  on
movements.  Previous blood data,  collected at the time of capture
and  instrumentation,  are  available.    It  is  anticipated  that
monitoring of these animals will  continue through 1991.  Recapture
and collection of a second blood sample as well as a urine sample
from these sea  otters  would provide  the opportunity  for further
physiological  and  toxicological  monitoring  of  these  animals.
Samples from the instrumented sea otters  would  be  of particular
interest because of the opportunity to relate  results to the known
history and continuing observations on the animals.

Many of the adult females  are expected to have  dependent pups in
the summer of 1991.  Capture and examination of  these pups would
provide an  opportunity to further investigate the  incidence of
physical abnormalities observed in 1990 captures.

Eastern portions of PWS were  not  directly oiled, and otters living
there  have generally been  considered a  valid control  for otters
found  in western PWS.   However,  given the critical  importance of
establishing reliable  baseline  values  for  Alaskan sea  otters,
capture  efforts   on sea  otters   in  a  second control area  are
necessary.

                           OBJECTIVES

The overall objective of this study is to evaluate bioindicators of
sea otters exposed to oil from the  EVOS.   Specific objectives are:

A.   To collect  blood  samples  from sea otters in western PWS and
     southeast Alaska.   Samples from western PWS will be compared
     to those from southeast  Alaska.   In western PWS, instrumented
     sea otters will be targeted because of their known history.

                               39

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B.   To  relate  blood analyses  on  sea  otters   in  western  PWS
     (instrumented otters)  with outcome  (survival and reproductive
     rates)  and  to compare  blood samples  collected in  1991  to
     previous samples collected on the same otters.

C.   To measure pre-weaning growth rates of  sea otter pups born in
     1991 in western PWS.

D.   To conduct physical examinations of  all  sea otters captured
     and  sedated,  for  evaluation of   health  and  detection  of
     developmental abnormalities.


                             METHODS

Capture activities will  be conducted in  June and July  1991  in
western PWS and southeast Alaska.

In  PWS,  adult  female sea  otters were instrumented with  radio
transmitters in the fall of 1989  and spring  of 1990, and sea otter
pups were  instrumented  in  the fall of  1990.   Blood  samples were
collected at the time  of capture.   Since  instrumentation, they have
been monitored to measure survival and reproduction rates  (for the
adult females). Due to the advantage of  obtaining blood samples on
individuals  of known history,  instrumented  sea  otters in  the
western Sound will be  targeted for sample  collection in the summer
of  1991.   An attempt will be  made to capture up to  30  of  these
otters.  If sufficient numbers of instrumented  sea otters cannot be
recaptured, additional non-instrumented  sea otters from western PWS
will be  captured  and sampled.   The  sea otters  will  be sedated,
blood collected by jugular venipuncture and, when possible,  urine
samples will also be collected.

Locations  of the  instrumented otters will  be  known  from ongoing
radio tracking efforts.   Capture  methods will include divers using
Wilson traps so that specific individuals  can be targeted.  Tangle
nets and dip nets  will be used as a supplementary  capture method as
needed.

Most of the adult females will be accompanied by a dependent pup,
which  will  also  be  captured  and  physically  examined  by  a
veterinarian  experienced in  handling  and  treating  sea otters.
Approximately  60  days after  the  initial  capture,  pups  will  be
recaptured, and weights and lengths again  taken to estimate growth
rates.    Previous studies  (Monnett,  unpublished data)  provide
information on pre-spill growth rates for comparison.

In southeast Alaska (Sitka control area), sea otters will  be caught
using tangle nets.  Adult  otters of  either sex will be targeted.
Animals will be sedated, physical examinations will  be done, and
blood and urine collected.

                                40

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Approximately 4 cc of whole blood will be put in a chemically clean
jar and  frozen for toxicology analysis.  Whole blood  (in a EDTA
tube) and serum will  be air-expressed to a qualified laboratory for
analysis  of  complete blood counts and  blood  chemistries.   Fresh
blood smears will be  made at the time of collection.  Urine will be
collected by expression of the  bladder and  analyzed  in the field
with  reagent  strips,  and for  specific  gravities and  sediment
levels.

Procedures for drugging the sea otters and collecting blood samples
will be as outlined in previous  study plans  (MM 6, 1989 and 1990).

Capture and handling techniques  will  be similar to procedures used
in previous  studies  in  Alaska  and  California.    For  veterinary
panels, blood samples will be sent to the same laboratory used in
1989 and 1990 NRDA studies; a subset of samples will be sent to a
second  laboratory,  located in  Alaska, for  comparison purposes.
Toxicology  assays will  be done  by  the same  laboratory  as  in
previous years, following established protocols from 1989 and 1990
studies.

Information  on  locations  of  instrumented  sea   otters  will  be
obtained  from  ongoing  telemetry  studies   on  these  otters.    A
clinical pathologist will be required for a interpretation of the
blood  results.    Mapped  data  on  shorelines  and  offshore  areas
affected by oil  will be  available for correlation with sea otter
capture locations and blood results.
                          DATA ANALYSIS

Blood values  (veterinary panels  and toxicology)  from southeast
Alaska  (control area)  and  western  PWS  will  be compared  in an
exploratory data analysis,  using  t-tests  to test for differences
among the two areas.   All variables will be  examined for normality
and  homogeneity  of   variance  and   transformed  as  appropriate.
Toxicology  values of blood  samples from  western  PWS will be
compared to values for the samples  collected  in 1989-90 using a
paired t-test.  Additionally, for samples from western PWS, blood
values will be related to the history and outcome  of the individual
sea otter.   For  example,  values of sea otters that survive through
the end of 1991 will be  compared to those of otters that die with
chi-square contingency table analysis.  These types of comparisons
will also  be  used to relate  outcomes to  specific locations  (and
degree of oiling thereof) where the otters have been residing.
                                41

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                              BUDGET
Salaries
Travel
Contracts
Commodities
Equipment

Total                      $ 88.4
                               42

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MARINE MAMMAL STUDY 66

Study Title:   Assessment of Pathological Processes and Mechanisms
               of Toxicity  in  Sea  Otters that Died Following the
               EVOS

Lead Agency:   FWS
                           INTRODUCTION

Following the  EVOS,  a massive effort  was  undertaken to capture,
clean,  and medically treat  sea otters  exposed  to crude  oil.
Following   the  spill,   329   sea   otters  were   brought   into
rehabilitation centers in Valdez and Seward. Approximately half of
these  animals  died  during rehabilitation,  a few were sent  to
aquaria, and the remainder were released to  the natural environment
in August,  1989.   Approximately  18  million dollars were spent by
Exxon to  rehabilitate affected otters.  Studies  on released sea
otters  are  providing  evidence  that a high percentage  of  these
animals may have  died following release (Monnett  et al.,  1990).
There  is  concern  that capture  and  rehabilitation may  not  be an
effective alternative for preservation  of the sea otter population
following exposure to crude oil.

The subset of animals  that died in captivity should  provide crucial
information  regarding  mechanisms  of   toxicity  associated  with
exposure to crude  oil and pathological  processes that caused death
following contamination  with  this toxic substance.   Analysis of
data  from  these  animals  will  provide  critical  information  to
determine  if  rehabilitation  is  a  useful alternative for  the
preservation of sea otter populations  exposed to crude oil.

Although numbers of recovered  carcasses were highest in the months
immediately following the oil spill, efforts to recover sea otter
carcasses from PWS have continued through 1990 and  are planned for
1991. Recovered carcasses may provide valuable clues to the factors
involved in the death of these animals. Work conducted under this
study will continue efforts that have been ongoing since the spill.
                            OBJECTIVES

A.   To determine the efficacy of sea otter medical treatment and
     rehabilitation as a viable method for the restoration of the
     Alaskan sea otter population following exposure to crude oil.

B.   To evaluate chronic effects of residual oil in the environment
     through examination of  sea  otter  carcasses recovered in the
     oil spill zone in 1991.   Work conducted under this study will
     continue efforts that have been ongoing since the spill.
                                43

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                           METHODS

A.  Sea Otters from Rehabilitation Centers

     In  the six  months  following  the  EVOS,  pathologists  from
     Environmental Protection Agency and the Armed Forces Institute
     of  Pathology were  on  site  and performed  complete  gross
     necropsies on all  sea  otters that  died at  rehabilitation
     centers.   Histopathology  of samples  collected from  these
     animals  will  be  integrated  with  the  clinical  records,
     hematology,   clinical   chemistries,   and   chemical  residue
     analyses.   The specific objectives of this study are:

     •    to describe the gross  anatomical and histopathological
          lesions  in  sea otters that  died  at  rehabilitation
          centers;

     •    to develop  a  model to  describe  the toxic  effects and
          pathological processes  that  caused death  in sea otters
          following exposure to crude oil;  and

     •    to test whether the necropsy,  histopathology, toxicology,
          and hematology results are statistically related to the
          geographic location of capture, severity of oiling, date
          of  exposure,   duration  of  exposure, or  the  changing
          composition of oil.

B.  Recovered Sea Otter Carcasses

     In  1991,  carcass  recovery efforts will be continued.   Ages
     will be determined for  recovered carcasses.   Necropsies of
     these  carcasses,  with  sampling  for   histopathology  and
     toxicology, will  further our understanding  of pathological
     processes associated with long-term exposure to residual oil
     in the environment.  In addition,  1991  studies of sea otter
     foraging  behaviour  will  determine  prey  composition  and
     hydrocarbon levels  of  prey  for  sea otters in western PWS,
     which can be related to body hydrocarbon levels.


                              BUDGET

Salaries                      $   0.0
Travel                          20.0
Contractual Analysis            22.0
Administrative Support            5.0
Equipment                       14.0

Total                         $ 61.0
                                44

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MARINE MAMMAL 6H

Study Title:   Sea  Otter Damage  Assessment  Studies:    Database
               Management and Data Analysis

Lead Agency:   FWS
                           INTRODUCTION

Two years  of oil spill response  efforts and NRDA   studies have
produced large amounts of data on sea otters affected by oil.  To
date,  most  of these data  have  had only a  preliminary analysis.
NRDA studies on sea otters, with the full or part-time involvement
of over 10 scientists,  will continue to generate new data in 1991.
                            OBJECTIVES

The objectives of the work outlined in this proposal are:

A.   To provide database  support,  including  data entry,  editing,
     and record management, for ongoing sea otter studies.

B.   To support statistical analyses and write-up of data generated
     in previous and ongoing sea otter studies.


                             METHODS

The objectives  of this proposal  will be  met  by support  of one
scientist, one database manager, and one biotechnician.  All three
individuals will be full time.  The majority of their time will be
spent in Anchorage working on data; however, a portion of the time
of all three will  be spent in the field assisting with 1991 damage
assessment  studies,  as needed.   Studies  or data  sets  requiring
support and analysis are listed below.


                          DATA ANALYSIS

1.   Morgue/carcass recovery:  Almost  900 carcasses were recovered
     within 6 months  of the oil spill, and  recovery  efforts are
     still  continuing.    Carcassses  are  maintained  in  frozen
     storage.  Necropsies  have been done on  most animals and, as
     feasible,  samples collected  for  histological analysis and
     toxicology.   Additionally,   teeth  have  been collected and
     submitted for aging  and reproductive  tracts of  females have
     been examined.   Identification numbers  are now  being cross-
     checked and  all  data compiled in one database.   Biological
     samples  collected are  stored or  shipped,  as required for
     analyses.

                               45

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2.   Sea otters from rehabilitation centers:  Following the spill,
     329 sea otters were captured in the oil spill zone and placed
     in otter  rehabilitation  centers  in  Seward and Valdez.   One
     hundred and seventeen of those otters died in the centers.  In
     addition, sea otters that were not considered able to survive
     in the wild following the rehabilitation process were sent to
     aquaria,  and  their health is being  monitored.   Records  on
     condition and health,  medical  treatments, blood collections,
     and behavior were kept for all otters.   A thorough study is
     ongoing to  evaluate pathological processes contributing  to
     death following exposure to oil,  and evaluating the success of
     the rehabilitation  effort  (see  MM 6H) .   Clinical  data  are
     currently being coded  by veterinarians who worked  with  the
     otters on a  daily basis, and this  information will be combined
     with  histopathology,   clinical  pathology,  necropsy,   and
     toxicology information.  Portions of this  database are not yet
     in digital  format,  and thus support  is required  to organize
     and  maintain  all  records  on   the   sea  otters  from  the
     rehabilitation centers, and to provide data as required to the
     cooperating pathologists involved in  this study.

3.   Blood data:   Since the  oil spill,  blood samples  have  been
     collected  on  approximately  200  sea  otters  in  PWS  (not
     including sea  otters  that had  blood  samples  drawn at  the
     rehabilitation centers).   Additional blood samples  will  be
     collected in the summer of  1991.  Analyses of these data will
     include  relationships  between  blood   panels   (CBC's   and
     chemistries) and toxicology (hydrocarbon  levels),  geographic
     locations,  and reproductive and  survival information on the
     otters.

4.   Toxicology data:  Tissue  samples from carcasses (depending on
     condition)  have been  collected  for analysis of  hydrocarbon
     levels.   Additionally,  blood  and   fat   samples  have  been
     collected from live animals caught in PWS since  the fall of
     1989.    Several thousand samples are now in frozen storage
     pending analysis  of hydrocarbon  levels.   Results  have  been
     received for approximately 150 tissue  samples;  250 more are
     currently being tested.  Analysis of the  toxicology data set
     will require input from a biostatistician and toxicologist as
     well as direction from the scientists who have been involved
     in the  studies to date.   Prioritization will  be  done  on
     additional samples to submit for analysis. Relationships with
     other   information  available   on   the   animals   will   be
     investigated.

5.   Survey data:  In 1989,  helicopter surveys were done on the KP
     and the KAP to determine  sea otter abundance and distribution
     prior to the arrival of oil (April and May),  and again after
     the oil had  affected  these areas  (August and  September).
     Analysis of these data will be undertaken.


                                46

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                              BUDGET

Salaries                      $107.6
Travel                          11.3
Contract                        10.0
Commodities                      2.5

Total                         $131.4
                               47

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               TERRESTRIAL MAMMAL  INJURY ASSESSMENT

Terrestrial mammals are an important part of the ecosystem in the
area affected by the EVOS.  A wide variety of species are present,
many of which use intertidal habitats that were heavily impacted by
oil.  They are important to humans for recreational viewing, sport
and subsistence hunting, and commercial and subsistence trapping.

In the  1989  damage  assessment plan,  14  species  were selected for
study  from  a  total  of 19  species  that  were  identified  as
potentially being impacted by the  oil spill.   In  1990, studies were
continued  for four species:  deer,  mink,  river otter,  and brown
bear.  A literature review on the  importance of intertidal habitat
use by   black bear was also done.   During the  coming  year,  work
will continue on river otter and brown bear only.

River otter  work will  continue to  examine  lethal  and sublethal
injury within the  oiled and unoiled study  areas established last
year.    This  includes  examination  of  animals found  dead  and
assessment of oil impacts on populations, food habits and habitat
use.   In addition, several aspects of sublethal injury  will be
investigated  on a  broad scale by  expansion  of  data  collection to
oiled and  unoiled  areas of  PWS that are outside  the established
study areas.

Brown bear investigations  will be  limited  to  monitoring female
bears  that  were  radio-collared  during 1989  and  1990.    Any
mortalities  will  be  noted   and  the  cause of  death  will  be
investigated.
                                48

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TERRESTRIAL MAMMAL STUDY NUMBER 3

Study Title:   Assessment  of the  effect  of  the  EVOS on  River
               Otters in PWS

Lead Agency:   ADF&G


                           INTRODUCTION

River  otter  (Lutra  canadensis)  populations  in  PWS  rely  on
intertidal and subtidal environments for  food.  Studies of similar
coastal populations in southeastern Alaska documented that marine
fishes,  crabs,   and  other  invertebrates  dominated  food  habits
(Larsen  1983,  Woolington 1984).   Because critical  habitats for
otters were heavily contaminated by oil,  otter populations are at
risk by  direct  contact with  oil or by  environmental  changes to
other  components of their habitats  in  response  to oil.   Data
regarding population density  prior  to the oil spill  are lacking,
but otters were  probably abundant.   The  goal  of  this study is to
determine  if the VOS  had  measurable  effects   on  river  otter
populations.  The approach is  (1) to examine carcasses to determine
direct effects  of oil,  (2)  compare pre-  and  post-spill  dietary
information  from scats,  (3)  continue  comparison of  population
density and various biological  aspects between oiled and control
study areas, and  (4)  relate biological aspects of river otters in
different areas  of PWS  to the  degree  of oil contamination and
environmental impacts  identified for these  areas in  other oil-
impact studies.

This study will employ extensive sampling of river otters through
live-capture techniques throughout PWS.   Work already accomplished
in the two intensive study areas (Esther Passage control area and
Herring Bay/Lewis Bay oiled-area) has provided data on body mass-
length  relationships  and  blood  values  for  otters.   Extensive
sampling will provide data on these relationships  in all components
of the otter population.  Additionally, the study will relate this
data to  varying  levels  of  oil contamination and environmental
impacts,  by sampling  study  sites  established  by  other  impact
studies (e.g., intertidal invertebrates and fish).  A  larger sample
size of otters than can be  obtained  from  the intensive study areas
is necessary to  identify  population  level impacts  for river otters
in PWS.

Continued work in the intensive study areas will monitor changes in
population levels, activity patterns, and home range size of the
previously  documented  otter  populations.   These  data will be
related to 1990  data to  identify trends  that  may be important to
proper interpretation of data from  the  extensive sampling effort
and for long-term trends.   This work will continue to use radio-
telemetry,   radioisotope   labeling   of   feces,   home   range
determinations,   and activity patterns to  provide parallel data.

                                49

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Additionally, animals will be live-captured and released throughout
the summer to provide comparable blood and body measurements with
the extensive program.


                            OBJECTIVES

Direct Effects

At.   Determine cause of death for river otters recovered from oiled
     areas via necropsy and histopathological procedures.

A2.   Test (a = 0.05)  for  higher hydrocarbon levels in river otters
     in oiled versus unoiled areas.

A3.   Determine  sub-lethal  effects  of  exposure  to  oil  on river
     otters.

Population Change.

Bj.   Estimate population sizes  of river otters with 10% of the true
     value 95%  of  the time, on representative  oiled and control
     study areas using mark-recapture methods, and test  (a = 0.05)
     for lower population levels in oiled versus control areas.

B2.   Estimate the rate of fecal deposition for river otters within
     10% of the true value 95%  of the time. This rate will be used
     as an index to  population size to test  (a  = 0.05)  for lower
     rate of deposition in oiled versus control study areas.

B3.   Test   (a  = 0.05) for lower survivorship of river  otters in
     oiled versus control study areas.

Food Habits

B4.   Test (o = 0.05)  for differences in food habits of river otters
     before and after the oil  spill on the oiled study area.

Bs.   Test (a = 0.05)  for differences in food habits of river otters
     on oiled and control study areas.

Habitat Use

B6.   Test  (a  =  0.05)  for   differences  in  activity  patterns
     (foraging) of  river otters between  oiled  and  control study
     areas.

B7.   Use home range  size and use patterns to test (a =  0.05)  for
     differences in habitat  selection in river otters between oiled
     and control study areas.
                                50

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                             METHODS

Methods used in 1990 will be continued in 1991.  Trapping areas for
the extensive live-capture program will be selected to provide data
from  differing  levels  of  oil contamination  and  to allow  the
greatest  use of site-specific  data from other  appropriate oil-
impact  studies.   The intensive study  areas will  be  utilized to
provide  continued   data  on trends  in  otter  populations  and  to
provide continuity  for  interpretation  of data  from the extensive
program.

The following are methods for collecting data by objective.

Direct Effects

Aj.   Necropsy  and   histopathology  procedures  will be  performed
     according to standard protocols.

A2.   Hydrocarbon protocols are established.  No additional animals
     will be collected  but hydrocarbon  and histological samples
     will  be  taken  from   all suitable carcasses  that  become
     available.

A3.   River otters will  be  live captured at  latrine sites in both
     study areas and at  pre-selected areas of PWS.  The techniques
     will be the same as used in 1990.  The modified Hancock live
     traps  and drugging boxes to  hold  otters,  as described  by
     Melquist  and   Hornocker   (1979),  will  be  used.      Weather
     permitting, traps will be monitored morning and evenings, and
     traps will be  equipped with a trap transmitter  that signals
     a sprung trap.   Otters will be  held only as long as necessary
     to obtain body measurements,  draw  a blood sample, and extract
     a  premolar for  age determination.    Animals  will then  be
     released at their original capture site.

     Standard procedures will be used to collect and process blood
     in the field.   Obtaining blood  values and morphometrical data
     from  the  same animals  should increase  the  power  of  our
     analysis  and   allow  a  more  complete  understanding  of  the
     relationship between these values and  their relationship to
     oil contamination.

Population Change

B!. .  In May, river  otters will be  captured in both study areas for
     this objective.  These  animals will be surgically implanted
     with  a  standard  implantable   transmitter  encapsulated  in
     biologically inert  materials, and with radioactive isotopes by
     a licensed veterinarian.  Techniques for implantation of radio
     transmitters will  be  those utilized in 1990  and originally
     described by Woolington (1979). Animals will be held only as


                                51

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     long as necessary to complete the marking process and recover
     from surgery.  Animals will then be released at their original
     capture site.

     The  radioisotope   implants  will  provide   the  basis  for
     estimating the population  density  in the oiled and control
     study areas using a mark-recapture method.   Marking of feces
     will occur as the polylactic acid (PLA) tablets containing the
     isotope are absorbed by the body and the long-lasting tracer
     released (Crabtree et al. 1989).  Feces will be recovered from
     latrine sites to provide both early and late summer population
     estimates.    This  mark-recapture  technique  was  employed
     successfully in 1990.

     A closed population model, employing the  radio transmitters to
     determine exactly how many  marked animals are resident in the
     study area while scats are being sampled, will be used.  Mark-
     recapture models for closed populations are well  established.

B2.   Data to assess the rates of fecal deposition as  a  means of
     estimating river  otter  population  size  will continue  to be
     gathered.   These  data will  be used to assess population trend
     and habitat use patterns in the intensive study areas.

B3.   Estimates of survival will depend on data obtained from otters
     instrumented with radio transmitters.  Each transmitter is
     equipped with a  "mortality mode" so the fate  of individual
     study animals can be determined.   Data collection  for this
     objective will coincide with data collected for objectives Bj,
     B6,  and  B7.

Food Habits

B4  and B5.    Food habits of river otters  will be  described from
     prey remains in their feces.  Such procedures have been used
     successfully in past studies of these  species  (Gilbert and
     Nancekivell 1982).  A large sample of scats has  been gathered
     but those scats  gathered  for  objective  B2  and  B3  will be
     preserved  and  used  if  additional food habit  analyses are
     necessary.  Laboratory analysis of prey remains in feces of
     river otters will follow procedures outlined  by  Bowyer et al.
     (1983).

     Because of  differential  digestibility of prey  and variable
     rates of passage  through the gut, volumetric  measures of prey
     remains in mustelid feces are meaningless.  Consequently, the
     analysis will be confined to the occurrence of  prey items in
     latrines and will  be expressed in terms of percent of latrines
     with food items,  and percent of total food items (Bowyer et
     al. 1983).   To  ensure  that subsamples from a latrine are
     representative of that site,  all feces from that site will be


                                52

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     mixed  and a series  of subsamples  (about  the volume  of an
     individual  scat)   will be  drawn  and analyzed  separately.
     Sampling  will continue until  the function  between number of
     prey  items  and number  of  samples becomes asymptotic.   All
     latrines  included  in  the analysis,  however,  will contain at
     least five scats per sampling period.

     Because  sample  variance is unknown,  it  is  not  possible to
     specify the total number of samples necessary  to describe food
     habits  adequately  at this   time.     However,   monitoring
     reduction in variation of  the mean was addressed in 1990 by
     increasing sample size (of  latrines)  for important food items
     to ensure that  all proportions are estimated within 0.05 of
     their true value 95%  of the time (Kershaw  1964:29).  In the
     control area, 113  latrines are established and  in the oiled
     area there are  131 sample  sites.   Additionally,  a sample of
     scats excess to the food habit studies will be submitted for
     hydrocarbon analys i s.

HABITAT USE

B6.   Otter  activity  will  be monitored  by recording  the apparent
     activity  pattern  when the radio  signal  is  first  picked up
     during  telemetry  relocations.   In  1991,  emphasis will be
     placed  on obtaining  visual  observation  of  otters  in  the
     intensive study areas  to  obtain parallel data  on foraging
     areas and durations.

B7.   Habitat  data for  description of  the two  study  areas  was
     completed in 1990.  Data on home range and habitat selection
     of individuals will be  collected daily, weather permitting, by
     monitoring  telemetered animals.    Radio  tracking will  be
     conducted from a small boat, and the  entire coastline of both
     study  areas will  be  surveyed.    Because  river  otters  are
     distributed  immediately along coastal areas (Larsen  1983),
     telemetry  "fixes"  will   be   made  over   relatively  short
     distances, and multiple "legs" can be used in triangulation.
     Consequently, error polygons  should be small and biases from
     animal  movements  during  triangulation  will  be  minimal.
     Starting time of telemetry surveys will be randomized each day
     to help minimize any  bias  from diel activities  of otters on
     estimates of home range size and habitat selection.  Further,
     aerial  telemetry  may  be  conducted  if needed to determine
     locations of individuals  that cannot be  located  by boat.
     Telemetry transmitters will  be  equipped  with  a mortality
     signal that will allow the speedy recovery of dead animals.
     The recovery of isotope-labeled scats from  latrine sites will
     also confirm individual home  ranges  determined  by VHF radio
     telemetry.

Methods for analyzing data  are detailed below for each objective.
                                53

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Direct Effects

Aj.   A cause of death will be assigned to each river otter carcass
     based  upon  necropsy   report  and  lab  analysis  of  tissue
     specimens.   Hydrocarbon  levels  will be  presented  for  all
     usable samples.

A2.   A one-tailed  Z test for proportions  (Snedecor  and Cochran,
     1980) will be used to test this hypothesis.

A3.   Blood samples and standard body measurements were taken from
     otters live captured in 1990.   Differences in selected blood
     values of otters from the  oiled and nonoiled study areas will
     be tested  with multi-response permutation  procedures using
     "Blossom"  statistical  software   (Biondini   et  al.  1988,
     Zimmerman  et  al.  1985).     Regression lines  of length-mass
     relationships  will  be compared  according  to Neter  et  al.
     (1985).

POPULATION CHANGE

Bt.   Analysis for  river  otters will follow methods  described by
     Seber  (1982:;  120-121)  for  sampling a  closed population with
     replacement.  Population size and 95% confidence  intervals for
     both control and oil affected areas will be estimated.  A one-
     tailed Z statistic will be used to determine if the population
     density is lower  in the oiled area versus the control area.
     This test assumes that the population estimates are normally
     distributed and have equal variance  (Seber 1982: p 121-123).

B2.   Differences  in rates   of  scat  deposition between  oiled  and
     control study  areas will  be tested  (a =  0.05) with a single
     factor covariance analysis model (Neter  et  al.  1985: 848) .
     The response, variable will be rate of  scat deposition and the
     covariate will be the number of latrine sites  (to control for
     any differences in population size between study areas).  Main
     effects will include oiling and months of study.   Since a one-
     tailed hypothesis is being tested with  regard to the oiling
     main effect, the critical region for this section of the ANOVA
     table will be one-tailed.   If variances are not homogeneous,
     either a ranked ANOVA procedure will be employed or the data
     will  be  transformed  to  obtain  homogeneous  variance  or
     normality.

B3.   Estimation and  analysis of survival  distributions for radio
     marked individuals  will follow procedures of Pollock et al.
     (1989).  This method controls for censored observations due to
     transmitter  failure,   animals  leaving the  study  area,  and
     individual  animals  living longer  than  the  study  period.
     Depending  upon the  structure  of data, we will  use either  a


                                54

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     parametric likelihood function or nonparametric Kaplan-Meier
     procedure  coupled  with   log-likelihood   test  to  examine
     differences (a = 0.05)  in  survivorship  (by  sex and age class)
     of  individuals  inhabiting  the  two   study  areas.    Model
     assumptions include a random sample of  animals, that survival
     times  are  independent  for  different  animals,  and  that
     censoring mechanisms  are  random (Pollock  et al.  1989).   An
     additional year of sampling may be necessary to  obtain a
     sample size large enough to make valid comparisons between the
     oiled and unoiled areas.

FOOD HABITS

B4  and  B5.   Statistical  analysis will include only  food  items
     that compose at  least  10%  of the diet.  Comparisons of food
     habits, pre- and post-spill, between oiled  and control areas,
     and among months will be made with the Quade test, including
     multiple comparisons of food items (Conover 1980:296-299).

HABITAT USE

B6.   It is hypothesized that if availability of forage species in
     the subtidal zone were reduced due to oil,  otters would spend
     more time foraging to obtain a diet equivalent to that in the
     control area.  Additionally, changes in the density of otters
     in the  two study  areas  could  influence  activity patterns.
     Simultaneous  reductions  in  otter  populations  and  forage
     species  could  result  in  change  in  individual  activity
     patterns.

     Differences in activity of river otters (stratified by sex and
     age  class)  between oiled  and  unoiled study areas will  be
     tested (a = 0.05) with a two-tailed Mann-Whitney test (Conover
     1980: 216).

B7.   The procedures of Swihart  and Slade (1985a,b) will be used to
     correct for auto-correlation among home range locations and to
     determine  the  time  interval  to achieve independence  of
     observations.  An adequate number of relocations to assess the
     seasonal home  range of an  individual  will be  determined by
     obtaining an asymptotic relationship between home range size
     and increasing  number  of  relocations.   Once the proper time
     interval and sample size have been determined,  the method of
     Dixon and Chapman  (1980) will be used to calculate 25%, 50%,
     75% and 95% isoclines of home range use.

     Isoclines of home range use will be overlayed on detailed maps
     of coastal habitats. The 95% use isocline will be employed to
     determine  the  habitats available  for  a  particular animal.
     Proportional weighing  by  25%, 50% and 75% isoclines within
     each habitat will  determine  use.   Thus,  habitat use  and
                                55

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     availability will allow a determination of habitat selection
     for each telemetered individual.  Testing for differences in
     habitat selection (rather than use) between oiled and control
     areas is  essential  because a difference  in  habitat  use may
     occur as  a  result of differential availability of habitats
     independent of  effects  of oiling.   A knowledge  of  habitat
     selection by river otters is  essential for extrapolating from
     our study areas to  effects on habitat  oiled in other areas.
     Consequently, habitat selection  (by sex) will be inferred from
     a significant difference  (P  < 0.05)  in use and availability
     matrices   compared    simultaneously   with   Hotelling's   T2
     statistic;  a  posteriori  comparisons  of  individual  habitat
     types will  be accomplished using Bonferroni multiple tests
     (Johnson  and  Wichern 1988:188).  Similarly,  comparisons of
     habitat selection in oiled and control areas will be made with
     a multivariate  analysis  of  variance  (MANOVA),  again using
     Bonferroni multiple contrasts.
                           BIBLIOGRAPHY

Biondini, M.E., P.W. Mielke, Jr., and E.F. Redente.  1988.  Use of
     a roller press to obtain cuticular impressions    of   guard
     hairs on acetate strips.  J. Mammal. 64:531- 532.

Bowyer, R.T., S.A.  McKenna and M.E.  Shea.  1983.  Seasonal changes
     in coyote food habits as determined  by fecal analysis.  Amer.
     Midland Nat. 109:266-273.

Conover, W.J.   1980.   Practical nonparametric  statistics.   John
     Wiley & Sons,  New York, 493pp.

Crabtree, R.L.,  F.G. Burton,  T.R.  Garland,  D.A. Cataldo and W.H.
     Rickard.  (in  Review)  Slow-release  radioisotope  implants as
     individual markers for carnivores.  J. Wildl. Manage.

Dixon, K.R.  and J.A. Chapman. 1980. Harmonic mean measure of animal
     activity.  Ecology 61:1040-1044.

Gilbert, F.F.  and  E.G.  Nancekivell.  1982.   Food  habits of mink
     (Mustela vision)  and otter  (Lutra canadensis)  in northeastern
     Alberta.  Can. J. Zool. 60:1282-1288.

Johnson,  R.A.  and  D.W.  Wichern.    1988.    Applied multivariate
     statistical analysis.  Prentice Hall, New Jersey,  606pp.

Kershaw, K.K.   1964.   Quantitative  and  dynamic ecology.  Edward
     Arnold, London, 1983pp.
                                56

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Larsen, D.N.  1983.  Habitats, movements, and foods of river otters
     in coastal southeastern Alaska.  Unpubl. M.S.  Thesis, Univ. of
     Alaska Fairbanks, 149pp.

Melquist, W.E. and M.G. Hornocker.  1979.  Methods and techniques
     for  studying and censusing  river otter populations.   Tech
     Report 78, Forest, Wildl. and Range Exper. Station, University
     of Idaho, Moscow, Idaho, 17pp.

Neter, J.,  W.  Wasserman and M.H.  Kutner. 1985.   Applied linear
     statistical  methods.   Richard D.  Irwin,  Homewood,  Illnois,
     1127pp.

Pollock, K.H.,  S.R. Winterstein and M.J. Conroy.  1989.  Estimation
     and  analysis  of  survival   distributions  for  radio-tagged
     animals.  Biometrics 45:99-109.

Seber, G.A.F. 1982.  The estimation of  animal  abundance and related
     parameters. Macmillan,  New York.

Snedecor, G. W.,  and  W.  G.  Cochran.   1980.  Statistical methods,
     7th ed. Iowa State University Press, Ames Iowa, 507pp.

Swihart, R.K. and N.A. Slade.  1985a.   Testing for independence of
     observations in animal  movements.  Ecology 66:1176-1184.

Swihart,  R.K.  and N.A.  Slade.   1985b.   Influence  of  sampling
     interval on estimates of home range size.  J. Wildl. Manage.
     49:1019-1025.

Woolington, J.D.  1984.  Habitat  use and movements of river otters
     at Kelp Bay,  Baranof Island, Alaska.   Unpubl.  M.S. Thesis,
     Univ. of Alaska Fairbanks, 147pp.

Zimmerman, G.M., H. Goetz, and P.W. Mielke, Jr.   1985.  Use of an
     improved  statistical  method for  group  comparisons  to study
     effects of prairie fire.  Ecology  66:606-611.


                              BUDGET

Personnel                    $ 122.1
Travel                         19.6
Contract                      165.9
Supplies                       39.2
Equipment                      30.5

TOTAL                        $ 377.3
                                57

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TERRESTRIAL MAMMAL STUDY NUMBER 4

Study Title:   Assessment of EVOS on Brown Bear Populations on the
               AP

Lead Agency:   ADF&G

Cooperating Agencies:  DOI, NFS, FWS


                           INTRODUCTION

Brown bears reside  along a section of shoreline  on  the southern
edge of the AP that was impacted by the EVOS.  Brown bears may be
exposed to oil by eating tar balls, grooming oiled fur, consuming
oiled carcasses, and as top level consumers, through accumulation
of toxins in  the  food  chain.   Bears in the  area  reproduce on an
average of  every four  to five  years  and may  live  25  years or
longer.  Effects of oil exposure may be immediate, or more likely
would occur over longer periods of time.  The effects of short term
exposure to high concentrations of petroleum hydrocarbons may not
become evident for many years.

Aerial surveys and radio-telemetry were used during 1989 and 1990
to  study  population density,  female mortality  and exposure to
hydrocarbons in an oiled area within Katmai National Park, and in
an unoiled  area  near Black Lake.  In 1991,  the  study  will focus
only on the continuation of  radio-telemetry to obtain additional
mortality information.


                           OBJECTIVES

A.   Test  the hypothesis  that  the  survival  (excluding hunting
     mortality) of female brown bears near oiled areas of  the coast
     of Katmai National Park are lower than  in other coastal brown
     bear populations that were not exposed to oil.

B.   Determine  the  cause  of  death of  dead brown bears located
     during monitoring  flights  in Katmai National Park.  Obtain
     tissues for hydrocarbon analysis if suitable to determine if
     death  can be  attributed  to the  physiological  effects of
     ingesting hydrocarbons.

                             METHODS

A maximum of  34  previously  radio-collared  brown bears will be
located during monitoring flights between den emergence  (May) and
den entrance (October).  Monitoring will be  conducted 3 to 4 times
per month during critical periods and twice per month during mid-
summer.   The  presence  or absence of dependent offspring will be
noted when possible.

                                58

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The  radio  transmitters  fitted  to females  were equipped  with a
mortality indicating mode.   When the animal  is motionless for a
predetermined period (usually 6  hours)  the  signal  transmits at a
slower (or in some cases, faster) interval.   When movement occurs
(as when the animal was resting but not dead), the signal returns
to normal from mortality mode.   During monitoring flights, bears
whose radios transmit on mortality mode will  be visually located to
determine if they are dead.  If visual location from the air is not
possible, a ground search will be conducted.  Survival rates will
be calculated using the Kaplan-Meier technique.

If accessible,  dead bears will be necropsied  to determine the cause
of death and suitable tissues will be collected for hydrocarbon and
histological analysis.
                              BUDGET

Salaries                   $  41.5
Travel                         4.7
Services                      28.1
commodities                    0.7
Equipment                      i.o

Total                      $  76.0
                                59

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                      BIRD  INJURY ASSESSMENT

The  EVOS  resulted in the  death of  a  large number  of  migratory
birds, especially  seabirds,  waterfowl,  and bald eagles.   In the
months  following  the  spill  it became  apparent  that  the  vast
populations of numerous bird  species that  inhabit  or utilize the
spill  zone remained  at risk to  direct  mortality,  as well  as
sublethal, long-term injuries.

.Fourteen studies were developed and conducted during  1989 and 1990
to document injury to migratory birds.  It was recognized early in
the process that it was  not possible  to study all the bird species
potentially affected by  the oil spill nor the full scope of effects
to any species.  Therefore, efforts were concentrated on studying
key species or groups of species where injury was most evident and
could be determined in a cost-effective manner.

Five  of  these studies  will  be  continued  in  1991.   Studies  on
peregrine falcons and passerines were not continued because it was
determined that all data pertinent to assessing injuries had been
gathered.

Continuing studies have been expanded and/or modified in response
to comments from reviewers and  the public.  The eagle study will
provide  information  on losses  to  breeding populations,  chronic
injury,  and  carcass recovery  rate.   The seabird colony  and
waterfowl surveys  will  provide  a means  to  compare  pre-  and post-
spill  populations  as  well  as  determine  recovery  rates  and
mechanisms  for  impacted species.  The  seabird colony  work will
emphasize documentation of injury to  murre  colonies.  The sea duck
study will  provide important  information on sublethal effects of
the spill on various species of ducks that feed in the intertidal
and  subtidal  habitats  affected  by the  spill.    Finally,  an
additional effort will be made in 1991 to more completely catalogue
and more  efficiently  store the  numerous bird  carcasses  that were
collected during  the spill response.   This will  facilitate the
future distribution  of  these  birds to  interested universities or
museums.
                                60

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BIRD STUDY NUMBER 1

Study Title:   Further Examination of Bird Carcasses  from the EVOS

Lead Agency:   FWS


                           INTRODUCTION

Following the  EVOS,  thousands of dead  birds  were recovered from
beaches  and nearshore  waters by  clean-up crews and  stored  in
freezer vans.  It  is  important that these birds are put to their
best scientific use.   Interest in  obtaining these birds has been
expressed by various museums and universities for use  in scientific
research and education.

Given the  difficulties that  field workers faced in identifying
large numbers  of heavily  oiled birds and managing the storage of
the  carcasses during  and after  the  field  operations,   it  is
necessary to re-examine and organize the many birds presently being
stored in the  freezer vans.  The storage system for the carcasses
will  be reorganized  for  quick  and  easy  retrieval  of specific
carcasses in the future.   The re-examination of the unidentified
birds, partial carcasses,  and refinement of some identifications
from a broad to a  more specific  category will serve to provide a
better   basis   for   future   disbursement   of   the  carcasses.
Additionally, data important to other studies will also be gathered
from carcasses as they are examined.


                            OBJECTIVES

A.   Re-examine carcasses  for  the  refinement  of bird numbers and
     refine identification from a broad to a more specific  level.

B.   Classify carcasses according to the amount and distribution of
     oil on the plumage.

C.   Reorganize the storage system for the birds to allow for quick
     and easy  retrieval of specific birds.

D.   Update log sheets with the best available information.

E.   Gather data that are of value to other bird  studies.
                             METHODS

Initially, the 9,000 carcasses in the Seward and Homer freezer vans
will  be examined,  followed by  the remaining carcasses  in  the
Kodiak/Alaska freezer vans and the Valdez freezer van (about 23,000
carcasses).

                                61

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The carcasses  in  the Seward and Homer  vans are stored  in totes
(4' x 4' x 3')  that will be  lifted by fork-lift  from the freezer
van, placed  in a pickup truck  and  transported to  the warehouse
facility,  where  they  will  be  thawed  and  inspected.   Bags  of
carcasses in the other three  vans are not stored in totes, but are
simply piled on  the floor.  These vans will be thawed  to allow
removal of the bags.

At the warehouse, totes of carcasses from the Seward and Homer vans
will be thawed  and the contents removed.  The following information
will be updated on log sheets for each carcass:

     (1)  taxa (to species level where possible);
     (2)  state of decomposition;
     (3)  proportion of plumage oiled;
     (4)  distribution of oil on plumage; and
     (5)  completeness of  specimen  material  (some  carcasses are
          represented by only a sternum or a wing).

In  some  cases, data  on  age  class and  other parameters will  be
gathered to assist other bird studies.   The bags of carcasses will
be repackaged,  as necessary,  and will retain their original number
and data  sheet.   After  examination,  birds will be individually
bagged (when possible),  returned to the  freezer van, refrozen in a
compact mass,  organized  and  stored  so  that  specific  bags  can be
quickly retrieved.   By this process, the inventory of the contents
of each bag in the Seward and Homer vans will be updated.

The  Kodiak/Alaska  Peninsula vans and  the  Valdez  van will  be
examined following the Seward and Homer vans.  Because of the way
the carcasses are stored, it will be necessary to thaw these vans
entirely to remove  the bags.  Additionally,  it  is  probable that
there are  too  many  birds in the Kodiak/Alaska Peninsula vans to
store on shelves.   It may be  necessary to store a portion of these
in the Seward and Homer vans if space is unavailable.


                          DATA ANALYSIS

Data collected during the  process of  carcass examination will be
recorded  on standard forms,  photocopied,  and entered  into  a
computer database for analysis.  Most analyses will focus  on number
of carcasses, species, and degree of oiling.

At the end of  the study, a report will  be prepared.  This report
will  provide  a  complete  and  comprehensive  description  of all
carcass   material   and   the   complete  results   of  analyses.
Additionally, photocopies of  all data sheets will be  provided as an
Appendix to the report.
                                62

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                           BIBLIOGRAPHY

Piatt, J. F., C. J. Lensink, W. Butler, M. Kendziorek, and D. R.
     Nysewander.  1990.  Immediate impact of the Exxon Valdez oil
     spill on marine birds.  Auk 107:  386-397.

Sanger, G.A.  1989.  Seabird surveys between Kachemak Bay and
     southern Kodiak Island, September - October 1989.  Unpublished
     report, U.S. Fish and Wildlife Service, Anchorage, Alaska.


                              BUDGET

Personnel                    $105.0
Travel and Other Costs         50.0
Contractual                   158.0

TOTAL                        $313.0
                                63

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BIRD STUDY NUMBER 2

Study Title:   Surveys to Determine Distribution and Abundance of
               Migratory Birds in PWS and the Northern GOA

Lead Agency:   FWS


                           INTRODUCTION

This study is a continuation of a similar study undertaken in 1989
and  1990  to  examine  whether the  EVOS  caused  a decline  in  the
distribution  and  abundance  of  waterbirds  in  the  waters  and
shorelines affected by the spill, including PWS, Kodiak Island and
the  northern  portion  of  Shelikof  Strait.   These  waters support
abundant waterfowl  and seabird  populations  throughout  the year
(Dwyer et al.  1976, Forsell  and Gould 1981,  Hogan and Murk 1982,
Irons et al. nd.,  Nishimoto and Rice  1987).  Potential injuries to
waterbirds from exposure to the EVOS include, but are not limited
to, death,  changes in  behavior, and decreased productivity.  Using
surveys by  small  boats, this project will  collect information on
the summer and winter distribution and abundance of waterbirds in
PWS.  These post-spill data will  be compared  to data collected,
using similar  methods,  in pre-spill  surveys  to determine whether
the  oil  spill   affected  and   continues  to  affect  waterbird
distribution and abundance in 1991.

This  proposal  describes  the boat  survey  work  that  will  be
accomplished in the third year of this study.  (The aerial survey
portion of  Bird Study No.  2  has  been discontinued.)   PWS will be
surveyed in March  and July  1991.   This field  effort will  be
conducted  in concert  with the Marine  Mammal  Study  No.  6 (Sea
Otters).  Surveys will not be conducted on Naked Island in Prince
William Sound, on the  southern Kenai  Peninsula or on Kodiak Island
waters in 1991.
                            OBJECTIVES

A.   To determine  distribution and estimate  abundance  (with 95%
     confidence limits) of waterbirds in PWS.

B.   To test the hypothesis that estimates of waterbird relative
     abundances, using new  and comparable  historic data,  are not
     significantly lower (a =  0.05) in oiled than non-oiled areas
     in PWS.
                                64

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C.   To  estimate  the long- and  short-term  trends  of populations
     that were determined in previous objectives to be reduced by
     the oil spill.


                             METHODS

A.   Boat Survey Sampling Methods

Damage Assessment Surveys.  Surveys will  be  conducted jointly with
the sea otter survey component of Marine Mammal Study No. 6 using
three 25-foot boats each manned with an operator and two observers.
Observers will record all  birds and sea otters within 100  m on each
side of the boat within  survey transects,  and whether the  animal is
in the  water,  on  land,  or in the  air.   The  survey  window will
extend approximately 40-50 m ahead of  and 100 m above the moving
boat,  but  will  be  extended  for  animals that  exhibited  strong
avoidance behavior when the boat  was  more than 50 m  away (e.g.
scoters, murrelets, harlequin ducks, harbor seals).  Surveys will
be conducted only when seas are less than 2 feet. Date and time of
survey,  and  environmental variables including  wind velocity  and
direction,   air   and water   temperature,  weather,  observation
conditions,  sea   state,  tide,  presence  of  oil on  water  or  on
shoreline,  and presence  of human activity will also  be recorded for
each transect.

A  stratified random  sampling design  using  shoreline,  coastal/
pelagic  and pelagic strata will be used  to meet Objectives A-C.
Surveys will be conducted in March  and  July  1991.   Fewer transects
will  be sampled  in March than  in July because  winter weather
conditions make it difficult to complete a longer  survey.

The  shoreline  stratum  was divided  into 742  transects used  in
surveys  by  Irons  et  al.  (1988, nd)  (see  Pre-Oil Spill Surveys
below).  For the  March 1991 survey,  the same 100 randomly selected
transects  (covering approximately  13% of the  shoreline)  used in
March 1990  will be surveyed.  The July 1991 survey will include the
same 212 transects (covering approximately  30% of  the shoreline)
sampled in June,  July and August 1990 surveys.  These include 187
transects  randomly  selected  to  be surveyed  in  1989,  plus  25
additional  transects randomly  selected   from  the   population  of
transects surveyed by Irons et al.  (1988, nd) in 1984.

The shoreline stratum includes all water within 200  m of shoreline.
Transects will be surveyed by travelling 100 m offshore, parallel
to the coast,  at  5-10 knots.  One observer will record all animals
seen between the  coast and the boat while  the other  will record all
animals between 100-200 m offshore.

Pelagic  and coastal/pelagic  strata consist  of plots  of  water
delineated by 5-minute intervals (latitude and longitude) on NOAA

                                65

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charts.   Forty-six  of  206  coastal/pelagic  plots and  25  of  86
pelagic plots randomly selected to be  surveyed  in June,  July and
August 1989 and 1990 will be  surveyed  in  July 1991.   The same 86
pelagic  plots  previously used  in all  surveys  and  the  same  29
coastal/pelagic plots used in March 1990 will be surveyed in March
1991.  Plots exclude any  water within 200  m of the coast.  The two
strata differ  in  that coastal/pelagic plots intersect  more than
approximately 1 nm  (nautical  mile) of  shoreline,  whereas pelagic
plots intersect less than 1  nm of  shoreline.  For plots that are 5
minutes wide (east to west),  two  north-south transect lines located
1 minute inside the east  and  west boundaries of the  plot will be
surveyed.   For plots that  are  less  than 5  minutes  wide  due  to
intersection with land, either one or  two transect lines will be
surveyed, depending on plot size.   In cases  where  a plot would be
very small, it was  combined with  an adjacent plot,  so  that some
plots contain three transect lines.

Transects in pelagic and coastal/pelagic plots will be steered by
a combination  of  compass heading  and LORAN-C coordinates.   Boat
velocity for pelagic and  coastal/pelagic plots will be higher than
for  shoreline  surveys,  ranging  from  15-20  knots, depending  on
observation conditions.

Pre-Oil Spill Surveys.  Two major survey  efforts  by  the FWS were
made prior to 1989.   Original  data from these efforts were located
for this study for pre- and post-spill comparisons.

The  first  effort was  a   series of  4  boat  surveys  conducted  in
March/April 1972,  July 1972, March 1973 and  August 1973 (Dwyer et
al. 1975).  These surveys randomly selected  approximately 13% of
transects in pelagic  and shoreline  strata in 1972,  and randomly
selected transects within subgroups of these strata in 1973  ("open
water" and  "coastal"  subgroups  within the  pelagic  stratum and
"outer exposed beaches",  "inner exposed beaches"  and "inner bays
and fjords" within  the shoreline stratum).   Observation methods
were comparable to those  used in  Damage Assessment Surveys, with
transect width 100 m on either side of the boat,  except that small
bays were included in the shoreline stratum,  and were surveyed in
their entirety as  part of shoreline transects.  Although individual
transects used in these surveys were different  from those used in
Damage Assessment Surveys,  methods  were  similar  and population
estimates can be compared.

During July and August of 1984 and 1985,  a complete survey  of the
PWS shoreline was conducted, using observation methods similar to
those used  for Damage Assessment Surveys (Irons,  Nysewander and
Trapp nd).The  shoreline  was divided into 742 transects.    (These
transects were subsequently  sampled  for the  shoreline portion of
Damage Assessment Surveys).   The western half  of the  Sound was
surveyed in 1984, and the eastern half was  surveyed  in 1985.  No
surveys of pelagic strata were attempted  in either 1984 or  1985.


                                66

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B.   Quality Assurance and Control Plans

To ensure that project design and procedures are followed, 1) all
crew members  will partake in  training surveys prior  to initial
surveys,  2)  one  person  on  each  boat will  be responsible for
maintaining consistent data collection  procedures, 3) standardized
forms will be used during data collection, and 4)  data forms will
be checked at the end  of  each  day to ensure the integrity of the
data.

C.   Information Required From Other Investigators

Shoreline and pelagic boat-based surveys in PWS will be conducted
in conjunction with sea otter  surveys  outlined in Marine Mammals
Study No.  6.   Field  data collection,  computer data  entry, and
quality control will  be performed by  biologists  and technicians
from both the Marine Mammal Project and the Marine and Coastal Bird
Project.

Post-stratification of  shoreline and  pelagic  transects  based on
presence or absence of oil will  be based  on data  compiled by the
Coastal Habitat Study, the Air/Water (Subtidal)  Studies, and the
Technical Services Study No.  3.  Oiling information was collected
by the Alaska Department  of  Environmental Conservation (ADEC) in
early summer  1989  (ADEC Summer  1989 Shoreline Assessment Data),
fall 1989 [ADEC Fall  1989  Shoreline Assessment  Data  ("Fall Walk-a-
thon")]  and spring  1990  [Multi-agency  Spring  1990  Survey ("SSAT
Survey")].  These 3 datasets will be used together to compile the
maximum extent of shoreline oiling.  The area of water covered by
oil was estimated  from a map  based  on ADEC aerial observations,
from a shoreline oiling map and from a NOAA HAZMAT hindcast model
of the movement of spilled oil  (J.A. Gait and D.L. Payton, National
Oceanic  and  Atmospheric Administration,   Hazardous  Materials
Response Branch, Seattle, WA) .   The  shoreline oiling dataset and
our estimated  area  of oil on  the water were  automated  onto FWS
Geographic Information  System (CIS)  using  Arclnfo  software, and
were used to produce datasets  describing  the extent of oiling in
each transect.
                          DATA ANALYSIS

Population estimates and variances  (Objective  A).   Estimates for
oiled and  non-oiled areas of PWS  (as defined by  "oil  on water"
datasets, above), as well as estimates for the entire Sound, will
be produced by adding estimates generated for each stratum within
a survey.  For the shoreline  stratum,  these will be computed using
a ratio  estimator  as follows (Sheaffer,  Mendenhall and Ott 1986:
131) :
                                67

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 Population estimate:  fy = rtx


 Variance:  V(tv)  = (f )2? (r)  = f 2 (^;
 Bound on the error or estimation (EE) :  EE = 2\Jv (ty)
 where    ty - population estimate for the shoreline stratum
           y

          r =
          YI = number of birds counted on the shoreline transect
          X( = area of the shoreline transect in km2
          rx = total  area of all shoreline transects in km2
      V (t ) = estimated variance of t
       V (r) = estimated variance of r
          N = total  number of shoreline transects
          n = number of sampled shoreline transects
          p. = mean area of all shoreline transects
The formulas will be the same for pelagic strata except that 1) Y;
will be estimated as  the  density of animals counted in transects
multiplied by  the  area of the  block sampled, and  2)  the finite
population correction  (fpc=(N-n)/N) will not be included.

Using  ratio  estimators  is appropriate  if  the  number  of birds
counted is positively correlated with transect length.  The extent
of  such  a correlation will  be determined.   Simple  totals  and
variances will be calculated  if  the correlation between counts  and
transect length is poor.

Statistical tests   (Objective B).   To examine whether oiled  and
non-oiled populations  changed in the same  way between the Irons
1984 shoreline survey  and surveys conducted after the spill,  the
change in population size  in oiled shoreline areas compared to non-
oiled  shoreline  areas  will   be computed  as follows  (after  log
transformation) for transects surveyed within a given  month  (July
or August):
                                68

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 Change in population = [(^-^2)^ - (Ri~R2^
 in oiled compared

 to non-oiled area
 variance of change - [ V*r(f •   ' f^>  +  v*r(T-
                             •  oiled               non-oiled


 where    f j  = estimated population total in 1984
          f2  = estimated population total in 1989 (or 1990)
          yl  = 1984 counts transects surveyed in 1984 and 1989 (or 1990)
          y2  = 1989 (or 1990) counts .from transects surveyed
                   in 1984 and  1989 (or 1990)
          w  = counts transects surveyed in 1984 only
          z  = counts transects surveyed in 1989 (or 1990) only
          x  = transect area for y (x) ,vf (x')  and z (x' )
               x+x
                       i       i   (s2     + R2 s2   — 2J? s      }
 Tr A >. /«   * \ _ vl r t    ""•      -^ \   \counts    1   tarea   &**\  \counts,area '
 Vox I T | ~ T^J — A  II —	— — — )	
     *A    /'      LX/»%^»^\    XT'                  /*5
                                 S     + -R 2S 2area
                                           V 4- V*  2

                                          ( *  *'  )
                       (n, + n2) (nj + n3)
                            (n, + n2)    (n, + n3)
The western half of PWS was  surveyed in 1984, and the eastern half
in 1985.  Transects  surveyed in 1985 were  not combined with those
sampled in 1984 because few  transects affected by oil were sampled
in 1985; this  meant  that variation due to  year surveyed could not
be distinguished from variation due to  oiling.  A  separate test
using  1985  data  could not  be  conducted  because  there  were not

                                 69

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enough  transects  sampled  in  oiled  areas  in  1985  to  perform
statistical tests.

The above formulas allow the use of transects that were sampled in
either pre- or post-spill surveys, as well as transects that were
sampled  in  both survey  periods.   T-values and their associated
probabilities can be derived from them.  If possible,  both oiling
definitions (shoreline and "oil on water") will be applied.

Two-sample  t-tests  will be performed  on datasets  consisting of
population estimates from each survey in a given month  prior to and
after the spill.  For example, population  estimates for all strata
combined  for  the  month of March 1972,  1973  and  1991 will  be
compared.

Post-stratification  of  PWS  into habitats for various species is
currently underway  using previously collected data on shoreline
types,   bathymetry   data  and   examination  of  each  species7
distribution.   Such stratification may  make statistical tests more
sensitive to  spill-related  population changes.    All  statistical
treatments may be revised after such stratification.

Maps  indicating  distribution  and  abundance  of  birds  will  be
produced for each survey to illustrate  differences between surveys
and oiled and non-oiled  areas.  Graphs of bird  abundance will be
produced and updated with each survey to show population trends and
differences.  Bird  density  and abundance estimates will also be
presented in tabular form.


                          BIBLIOGRAPHY

Dwyer, T.J., P.  Isleib,  D.A. Davenport,  and J.L.  Haddock.   1975.
     Marine bird populations in Prince  William Sound Alaska.  U.S.
     Fish and Wildlife Service, Anchorage, Alaska.   Unpublished
     Report, 21 pages.

Forsell, D.J., and P.J. Gould.   1981.  Distribution and abundance
     of marine  birds and mammals  wintering in  the Kodiak area of
     Alaska.  U.S. Fish and Wildlife  Service, Office of Biological
     Services, Washington,  D.C.  FWS/OBS-81/13.   81 pages.

Hogan, M.E., and J. Murk.  1982.  Seasonal distribution of marine
     birds in Prince William Sound, based on aerial surveys, 1971.
     U.S.   Fish  and  Wildlife   Service,   Anchorage,   Alaska.
     Unpublished Report.

Irons,  D.B.,  D.R.  Nysewander,  and  J.L.  Trapp.   1988.   Prince
     William Sound sea otter distribution.  U.S. Fish  and Wildlife
     Service,  Anchorage, Alaska.  Unpublished Report,   31 pages.
                                70

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	, 	  &  	.   nd.   Prince William  Sound waterbird
     distributions  in  relation to habitat  type.   U.S.  Fish and
     Wildlife Service, Anchorage, Alaska.  24 pages.

Nishimoto, M. ,  and  B.  Rice.   1987.  A  re-survey  of seabirds and
     marine mammals along the south coast of the Kenai Peninsula,
     Alaska during  the summer of  1986.   U.S.  Fish  and Wildlife
     Service, Alaska  Maritime National Wildlife  Refuge,  Homer,
     Alaska.  Unpublished Report, 79 pages.

Sheaffer, R.L.,  W. Mendenhall and L. Ott.  1986.  Elementary survey
     sampling.     Third  edition.     PWS   Publishers,   Boston,
     Massachusetts.
                              BUDGET

Personnel                     $141.0
Travel                          10.0
Contractua1                     30.0
Supplies                        33.0
Equipment                        6.0

Total                         $220.0
                                71

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BIRD STUDY NUMBER 3

Study Title:   Population Surveys of Seabird Nesting Colonies in
               PWS, the Outside Coast of  the KP,  Barren Islands,
               and Other Nearby Colonies,  with Emphasis on Changes
               of Numbers and Reproduction of Murres

Lead Agency:   FWS
                           INTRODUCTION

The 1989  EVOS  prompted resurvey of  seabird colonies in  PWS and
other areas westward  along the spill trajectory.  Most  of these
colonies were censused at least two and up to six different years
out of the previous 17 years prior to the  oil  spill.   Murres and
kittiwakes  on  one  nearby  colony  site,  Middleton Island,  were
censused  11  of  the  17 years  before the  spill.   Cliff-nesting
species such as  the black-legged kittiwake and common murre were
the primary emphasis of the 1989-90 censuses. Timing of egg laying
and productivity (numbers  of fledgling chicks) were also noted for
these species.    In 1990 the  major  effort  was placed  on replicate
counts of murres.  Semidi Islands and Middleton Island monitoring
continued as the main control sites for murres.

There are  approximately  320 seabird colonies,   not  including the
Semidi Islands,  that  occur within the  area affected by  the oil
spill.  Before the spill  they  contained about  1,121,500 breeding
seabirds  of  which 319,130 were  murres  (FWS, Catalog  of Alaskan
Seabird Colonies—Computer Archives 1986).   The  Semidi Islands
contained  an additional  1,133,000  murres  of  both species  (FWS
computer archives 1986).   Diving seabirds  are  known  to be easily
impacted by oil spills (King and Sanger,  1979).   In addition, these
species are long-lived and have low reproductive rates, thus making
any mortality of  adults a critical factor in these species' ability
to recover from  loss.

This study will continue this year to look at changes in numbers of
adult murres  at  the  breeding colonies  selected:    (1)  Chiswell
Islands,  (2) Barren Islands, (3)  Puale Bay/Cape Unalishagvak, and
(4) Semidi Islands.   Productivity  and  phenology will be measured
from  land-based   plots  in  the Semidis  and compared with  that
recorded similarly at  the  Puale Bay colony  to develop estimates of
productivity and phenology at the other colonies where land-based
plots are not feasible.

                            OBJECTIVES

A.   Determine whether the numbers  of selected  species of breeding
     colonial  seabirds  within  the  oiled  area have  decreased
     compared to numbers previously censused at these  sites.  Non-
     oiled nesting colonies will be surveyed as  a control.

                               72

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B.   Compare reproductive  chronology  and productivity for murres
     and kittiwakes  at colony  sites  within the oiled  area with
     those  found  at  nearby  colonies  in  the  GOA not  affected
     directly by the EVOS.


                           METHODOLOGY

This study will continue to look primarily at changes in numbers of
breeding adult murres at the previously mentioned sites.  In some
areas  there will  be  a secondary  emphasis on  counts  of  other
selected  species  such  as  black-legged  kittiwakes,  cormorant
species, and parakeet  auklets  if weather,  logistics,  timing,  and
geography allow.  Total counts are not feasible at large colonies
like the Semidi and Barren Islands and hence previously established
plots will be used of certain subcolonies.

Specifically,  the two strategies  used  in  1989  and  1990  will
continue to  be utilized:   (1) counts of adult  seabirds on plots
from  land-based observation points;  (2) counts  from boat-based
observation vantage points where land-based observations are not
possible.  If  plots or subdivisions are not possible, then total
counts or photography from boats will be the sole option.  Aerial
photography will not work at this time because the murre colonies
were highly  asynchronous,  and will not  stay on  the colony.   The
above strategies, in combination with the widespread distribution
and number of colonies to be examined, determined that the sample
plan would have two basic applications for 1991:

     (1)  A combination of  total counts and establishment/review of
          plots counted from boats will  occur at  colony sites like
          the  Barren   Islands  and  Chiswell  Islands  because  the
          colonies are much larger, in very exposed waters, have a
          poor history of censusing, and require counts from boats.
          Sample plots  were  established in 1989 and  1990 on the
          basis of accessibility and visibility.

     (2)  Land-based plots will be continued at the Semidi Islands
          because these colonies are  too large for total counts.
          Land plots are feasible and have  been used  for over 10
          years.   Sample  plots  were  previously selected  on  the
          basis of accessibility.

The  AP  murre  colonies  have  required  a  combination  of  both
applications in  the  past  and will continue to  do  so since some
portions of the colonies are visible from land,  but most aspects of
the colony required boat counts.

Colonies will be recensused using the  standard FWS methodology for
either  land-based  or boat-based  counts of  seabirds  (Byrd 1989;
Hatch and Hatch  1988  and 1989;  Irons et al.  1987;  Nishimoto and
Rice 1987) .  This  will vary depending  on the  topography of each

                                73

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area.  At least three replicate counts will be conducted, between
1000 and  1700  hours,  of colonies  or  plots after eggs  are laid.
These three replicate  counts will be on three separate days.  Plots
and  photographs  (using  6x7  cm format  cameras)  will   again  be
utilized for establishment of correction factors of total counts,
comparisons with past  plots,  and for evaluation of future recovery
or change.   Survey units  are subcolonies for cliff  nesters  and
islands for other species.

During boat censuses,  seas  must be less than 3  feet and rain should
not  be more than a  light drizzle.   At  least three  observers
including skiff  operator make the counts by  binoculars  from  the
boat.  Each observer counts each section of the cliff at least two
times and all counts  are compared  to  see  if sections  of the plot
were missed (differences  in  counts by two observers  cannot  be
greater than 5%) and need more replicate counts.

Nesting phenology and  reproductive  performance on land-based plots
will  be determined  by  viewing nests  at  regular  intervals  of
approximately  3  days.    Nest  sites  will  be numbered on  plot
photographs and/or drawings and then followed throughout the field
season.  Attendance of  adults, nest starts, and the  presence  or
absence  of  eggs  or  chicks   are  recorded.    For murres,  it  is
frequently not possible to see the contents of a site because the
birds remain motionless for long periods of time.  Thus distinctive
behavior (e.g.  wings held over the  back so that tips do not cross,
tail down, back slightly humped) is used to indicate that a murre
is incubating an  egg.  Because it is  possible to  misinterpret a
bird's posture,  we will use the convention that a site has to have
a bird in "incubating  posture" on at least three consecutive checks
to consider the site as having an egg.  In a similar fashion, wing
mantling  will  be  used  to  indicate that  a murre  has  a  chick.
However,  only   one  sighting  of wing  mantling  is  necessary  to
consider a murre to have  a  chick or to  be in a "brooding posture."
The  conventions  of  murre monitoring used by  the Alaska Maritime
National Wildlife Refuge will be used to resolve any questions of
interpretation.

Phenology and productivity data cannot be gathered as intensively
at areas where murre colonies  can only be reached and observed from
boats.   Instead,  phenology will be determined  indirectly  by the
change  in degree of  murre  attendance  at the cliffs  since murre
attendance is highly  variable on a daily  basis  before egg laying
and becomes more consistent after that.  As  in 1990, some portions
of the  rugged  islands will be  climbed occasionally  whenever sea
conditions permit a landing and portions of murre colonies will be
scanned for eggs or  chicks.   Productivity will be  evaluated  by
number  of chicks  present on  plots or  subcolonies  near fledgling
times.
                                74

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                          DATA ANALYSIS

The standard procedures and assumptions used by the FWS on colonies
in the Alaska Maritime National  Wildlife  Refuge are described by
Carton 1988  and Byrd 1989.   Several  key assumptions are:   (1)
plots, by  necessity,  are not random  and selection is  based on
accessibility; hence this study  makes  the assumption that counts
within plots are representative of the way the counts varied on the
entire colony;   (2) counts of plots or entire colonies from boats
are very difficult  for large colonies and replications of counts by
several observers on the same day and different days  illustrate the
need to refine the  accuracy and the variation recorded.  This means
that even counts of entire colonies are considered a  form of index,
but this study assumes that changes  in  these  indices represent the
changes occurring  in the  colony;   (3) counts  are unlikely  to be
normally distributed and are  more likely to be  skewed and clumped.
This type of data requires either very large sample sizes, or the
use of a non-parametric test, or the data needs to be transformed
logarithmically and then tested by the appropriate parametric test.
This transformation normalizes the data and is required for valid
application of statistical tests  on  small  sample  sizes (Fowler and
Cohen 1986, D. Robson pers.  comm.).

The standard  FWS procedures  mentioned prefer to compare trends
between years using numerous replicate counts where all plots are
censused  each  count  day  and  these  counts  are   replicated  on
successive days. The average of daily counts on the  Semidi Islands
will be used to  calculate a confidence interval for the estimate as
was done on the  Semidi Islands data in the past (Hatch and Hatch
1988; Hatch and Hatch 1989; Dragoo and Bain 1990).   At other sites
where there are fewer replicate counts, the procedure used in the
past, which was  usually an average of the available counts, will be
followed.

Data for 1991 will  be treated similarly to  1990 data  using standard
t-tests on logarithmically transformed data for all colonies except
the Barrens where  an  analysis of variance  for the  comparison of
change in  murre numbers  (also log  transformed)  was used for the
Barrens versus the Semidis between  1979 and post-oiling years.


                            BIBLIOGRAPHY

Byrd, G.V.  1989.  Seabirds in the Pribilof Islands, Alaska: trends
     and monitoring methods.   M.S.  Thesis, Univ. of Idaho,  Moscow,
     Idaho, 96pp.

Dragoo, D.E. and B.K.  Bain.   1990.  Changes in colony size,  and
     reproductive success of seabirds at the Semidi  Islands,  Alaska,
     1977-1990.    U.S.  Fish   and Wildlife  Service,  Homer,   Alaska,
     Unpublished Report.


                                 75

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Fowler, J. and L., Cohen.  1986.  Statistics for ornithologists.
     British  Trust  for  Ornithology,   BTO  Guide  No.   22,   Tring,
     Hertforre.  175 pages.

Carton, E.O.  1988.  A statistical evaluation of seabird monitoring
     programs at three sites on the Alaska Maritime National Wildlife
     Refuge.  Univ.  of Idaho, Moscow, Idaho.  Unpublished Report from
     contract with the refuge, 15pp.

Hatch, S.A. and M.A. Hatch.  1988.   Colony attendance and population
     monitoring  of  black-legged kittiwakes  on  the Semidi Islands,
     Alaska.  Condor 90:613-620.

Hatch, S.A. and M.A. Hatch.  1989.  Attendance patterns of common and
     thick-billed  murres  at  breeding  sites:     implications  for
     monitoring.  J. of Wildlife Management. 53(2):483-493.

Irons, D.B., D.R. Nysewander, and J.L. Trapp.  1987.  Changes in colony
     size  and reproductive  success of  black-legged kittiwakes  in
     Prince  William  Sound,  Alaska,  1972-1986.     U.  S.  Fish  and
     Wildlife Service, Anchorage, Alaska.  Unpublished Report. 37pp.

King, J.G. and G.A.  Sanger.  1979. Oil vulnerability index for marine
     oriented  birds.   Pp. 227-239  in  Bartonek and Nettleship eds.
     Conservation of marine birds of northern North America.   U.  S.
     Fish and Wildlife Service, Washington D.C.  319pp.

Nishimoto, M. and B. Rice.  1987.  A re-survey of seabirds and marine
     mammals  along  the south coast  of  the Kenai Peninsula,  Alaska
     during  the  summer of 1986.  U. S.  Fish and Wildlife Service,
     Alaska  Maritime  National  Wildlife  Refuge,   Homer,   Alaska.
     Unpublished Report. 79 pages.


                               BUDGET

Personnel                     $124.4
Logistics                      140.8
Equipment                       18.0
Miscellaneous
Supplies/Services               35.3
Travel/Per Diem                 17.0
Contractual                    194.5

Total                         $530.0
                                 76

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BIRD STUDY NUMBER 4

Study Title:   Assessing the Effects of the EVOS on Bald Eagles

Lead Agency:   FWS


                            INTRODUCTION

The  area  affected  by  the  EVOS provides  year-round  habitat  for
approximately  5000  adult bald  eagles  and seasonal  habitat for  an
estimated additional 2500 immature bald eagles.  An unknown number of
bald eagles from breeding areas in southcentral Alaska may also winter
in the spill area.

Bald eagles are closely associated with intertidal habitats that were
heavily  impacted  by the  EVOS.   Nearly  all  nests in the  spill area
occur within 100 meters of the beach where eagles commonly forage in
intertidal habitats on fish and marine invertebrates.   Eagles that
breed elsewhere,  but  spend  winters  in  the spill area, also  use  the
impacted intertidal habitats for foraging.

This  study  is  a  continuation  of  work  designed to  document  the
magnitude and duration of impacts to bald eagles caused by the  EVOS.
Estimates for the number  of  eagles occupying the spill area after  the
spill will be compared with historical data to identify changes in  the
population.  Nestling and adult bald eagles from oiled and non-oiled
areas will be monitored to estimate  survival rates,  distribution  and
exposure to oiled  areas, and determine causes of mortality.  Estimates
of acute mortality will be improved  through assessment of the number
of dead  birds  found  in  relation to the  number  of birds that were
killed, but never found.   Blood samples will be  collected to monitor
the health of eagles within the spill area.

Because  eagles mature slowly and  are  long-lived,  impacts to  the
population may not  be readily  apparent.   Furthermore,  the long-term
impacts of oil contamination on bald eagles are unknown.


                             OBJECTIVES

A.   Estimate numbers of resident bald  eagles such that the estimate
     is  within 10% of the  actual  size 95%  of  the time;  determine
     whether changes  in  population size  have  occurred in  the oil-
     impacted areas since 1982 and test whether  the change in number
     of  eagles  in oil-impacted  areas  is  different than  changes in
     non-oiled areas.

B.   To test the hypothesis  that survival  rates are the same for bald
     eagles in oiled and non-oiled areas.
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C.   Determine  the proportion  of  eagles that  die  on  beachfront
     relative to the  number  that  die in areas away from  the  beach-
     front .

D.   Determine the toxic  and sublethal effects of oiling on  eagles
     and eggs.


                              METHODS

Population Surveys (Objective A) .  Surveys of randomly selected plots
will be conducted from Malaspina Glacier to Cape Elizabeth  in early
May, following methodology discussed in Hodges et al.  (1984).   All
shorelines in each selected plot will be flown at an altitude of about
200  feet  and  an  airspeed of 90 to 100 knots  using  fixed-winged
aircraft.    Eagles  will  be  classified  as  either  white-headed  or
immature.   "White-headed" eagles will include  sexually mature  adults
and near-adults that have predominantly white heads.  This survey will
not directly  estimate the number of immatures,  therefore,  we  will
assume that ability to detect all age  classes  is equal for  birds in
flight, and a  ratio of  adults to immatures observed  flying will be
used to estimate the number of immatures.

Survival Studies (Objectives B).  During the winter,  food resources
for bald eagles are at the lowest availability of the year and  eagles
are presumably under the greatest nutritional  stress.   Mortality due
to inadequate food will most  likely  occur during the  winter period.
Furthermore, some, contaminants  stored in fat tissues  are mobilized
during periods of nutritional stress.  To estimate survival rates, 135
eagles (64 adults and  71 nestlings from oil and non-oiled areas)  were
tagged with radio transmitters.  Bi-weekly aerial flights will be made
to  relocate the transmitters using  standard  telemetry  techniques
(Gilmer et al.  1981) and to document eagle numbers,  distribution, and
mortality within the study area.  Dead  eagles  will be retrieved and
necropsied to determine the cause of death.  Survival rates will be
estimated using the  Kaplan-Meier (1958)  procedure (Pollock  et al.
1989).  Survival functions will be tested for significant differences
between eagles marked in oiled and in unoiled  areas,  and between age
classes.  Long-term monitoring will allow calculation of seasonal and
annual survival rates and a  better  interpretation of  the long-term
effects of oil  contamination  on  bald eagle  populations through
population modelling.

Carcass Recovery Study (Objective C) .  Data from the telemetered birds
in the survival study will also provide information on the number of
birds that die on the beachfront relative to the number that  die in
wooded areas where they are unlikely to be found.   This will provide
an index to estimate the total number of eagles killed by the EVOS in
1989 relative to the number of eagle carcasses recovered during 1989.
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Toxic and Sublethal Effects of Oiling (Objective D) .  All eagles found
dead will be  collected and necropsied  to substantiate the  cause  of
death and look for  signs of oil contamination.  Tissue samples from
the  collected specimens will  be  analyzed  for contaminants.   All
histopathology work will be accomplished through the FWS  National
Wildlife Health Laboratory.  All samples collected in the field will
be properly labelled and chain of custody procedures followed.

Blood  samples from birds  which  are caught  and  released  will  be
collected and analyzed to determine concentrations of hydrocarbons and
other contaminants associated with oil  contamination.  Approximately
equal numbers of bald  eagles will  be sampled  from oil and non-oiled
areas.   Blood  samples will  also be  analyzed for  standard  blood
chemistry profiles, which will help identify  sublethal impacts.  Blood
chemistry of eagles will be compared between  oiled areas and non-oiled
areas,  and  tested  (2-sample  t-test,   a =  0.05)  for  significant
differences.  Blood chemistry results will  also be interpreted by a
veterinary clinical pathologist.


                           DATA ANALYSIS

Population  surveys  (Objective A) .   Analytical methods   and  tests:
Surveys will be conducted using a random plot design,  as discussed  in
Hodges et al.  (1984) .  This survey technique will allow estimation  of
the  changes  in  numbers of  adult eagles  and occupied  nests  when
compared with the previous  surveys  of PWS in 1982 and 1989, trying  to
obtain a confidence interval  of  ±  10%.   It will be assumed that  no
major changes in habitat quality or quantity that may affect the
breeding population have occurred  since 1982, other than the  EVOS.
The following hypotheses will be tested (2-sample  t-test  or  analysis
of variance,  a = 0.05):  (1) that the number of adult  bald eagles  in
the entire  survey area in 1989,  1990  and  1991 is the same as the
number of adult bald eagles in 1982; (2)  that the number of adult bald
eagles within the oil-impacted area is  the same for 1982,  1989, 1990
and 1991; and (3) that the change in numbers of adult bald eagles  in
the oiled areas  is the same as the  change  in numbers in non-oiled
areas among and between years.

A parametric two-sample t-test (Steel and Torrie,  1960) will be used
which does not require equal variances  to test the above  hypotheses.
Analysis  of  variance  will  be   used   for  multiple comparisons.
Assumptions necessary  for  valid application  of  the  t-test  will  be
checked  (e.g., test for normality).   If assumptions are violated,
either an appropriate transformation or an equivalent non-parametric
test will be used.

Survival Studies  (Objective B) .   Analytical Methods and  Tests:   It
will be assumed that all eagles in the study  area have an equal chance
of being captured and that all transmitters  have a negligible effect
on the  eagles behavior  and do  not influence the bird's chance  of
survival.  Survival data will be analyzed using the methods of Kaplan

                                 79

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and  Meier  (1958)  which  accommodate  infrequent  visitation  (i.e.,
relocations)  of  birds, and  censusing of  lost  birds.   This is  an
appropriate method because it is expected that eagles will move  from
the  study area where  they cannot be relocated during every  survey.
Furthermore,  the  Kaplan-Meier  method  does not  assume constant
survivorship during the period of observation.

A Z-test (Bart and Robson,  1982) will be used to  test for significant
differences in survival rates between eagles marked in oiled areas and
eagles  marked   in unoiled   areas.     This  Z-test  requires   the
transformation of  the  survival rate and  standard  error  to normalize
its  distribution  and allow  use  of  a   Z  statistic  to  test   for
differences in survival rates.  The potential exposure of individual
radio-marTced  eagles  in  oiled  areas based  on  frequent, accurate
relocations  will  be  substantiated  allowing  a  more   appropriate
classification  of  eagles  into  treatment  groups   based  on   the
proportional  amount of time  they were located  in oiled  or  unoiled
areas.

Toxic and  Sublethal Effects  of  Oiling  (Objective  D) .    Analytical
Methods and Tests:   Blood samples  will be  collected  from  eagles
captured in PWS  and will  be  tested  for  significant  differences  in
levels of contaminants and blood characteristics between bald eagles
from oiled  and non-oiled  areas using a 2-sample t-test  (a = 0.05).
Assumptions necessary  for valid application of the t-test  will  be
checked  (e.g.,  for normality).    If  assumptions are violated,  an
appropriate transformation or an equivalent non-parametric test  will
be  used.     Information  on   blood  characteristics  will also  be
interpreted by a  veterinary clinical pathologist to access impacts on
bird health.

The spring population surveys will be conducted between April and  May,
1991.  The  radio-marked  eagles will be monitored bi-weekly  between
February and June  1991.   Dead  eagles will  be collected  as available
between February and June  1991.  Blood will be  sampled  between  late
August and October 1991.


                           BIBLIOGRAPHY

Bart, J. and  D.S.  Robson.  1982.  Estimating survivorship when the
     subjects are visited periodically.  Ecology 63:1078-1090.

Gilmer,  D.S., L.M.  Cowardin, R.L. Duvall, L.M. Mechlin, C.W. Shaiffer
     and V.B. Kuechle.  1981.  Procedures for the  use of aircraft in
     wildlife biotelemetry studies.  U.S.  Fish  and  Wildlife  Service
     Resource Publication 140.  19 p.

Hodges,  J.I., J.G. King and R. Davies.   1984.   Bald eagles breeding
     population survey of  coastal British  Columbia.   J.  of Wildlife
     Management.    48:993-998.


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Kaplan,  E.L.  and P. Meier.   1958.   Non-parametric  estimation  from
     incomplete  observations.    Journal   of   American  Statistics
     Association 53:457-481.

Pollock, K.H., S.R. Winterstein, C.M. Bunck, and P.D. Curtis.   1989.
     Survival analysis  in telemetry studies:   the  staggered  entry
     design.  J. of Wildlife Management 53:7-15.

Steel, R.G.D. and J.H. Torrie.   1960.  Principals  and procedures in
     statistics.  McGraw-Hill, New York.   481 p.


                               BUDGET

Salaries                      $ 83.0
Travel                          17.0
Contracts                      137.0
Commodities                     14.0
Equipment                        4.0

Total                         $255.0
                                 81

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BIRD STUDY NUMBER 11

Study Title:   Injury Assessment of Hydrocarbon Uptake by  Sea  Ducks
               in PWS

Lead Agency:   FWS

Cooperating Agency: ADF&G


                          INTRODUCTION

This  study  will  focus  on  the effects  of  petroleum  hydrocarbon
ingestion by harlequin  ducks (Histrionicus histrionicus),  Barrow's
goldeneyes   (Bucephala  islandica),  common   goldeneyes   (Bucephala
clangula), black scoters  (Oidemia  nigra),   surf scoters  (Melanitta
perspicillata), and white-winged scoters (Melanitta deglandi)  in PWS
as a result of the  EVOS.  PWS is a major wintering area for  these sea
duck species (Isleib and Kessel,  1973) .   It  is  also an  important
migration area for sea ducks in spring and  fall,  and a breeding site
for  resident  harlequin ducks  during  the  summer  (Hogan,  1980).
Harlequin ducks in particular, because of their resident status and
intertidal foraging habits, are considered substantially at risk to
effects of the EVOS (King and Sanger,  1979).  Goldeneyes and scoters,
although migratory, are also at risk because  of their intertidal and
subtidal foraging habits.

The six sea duck species included in this study are heavily  dependent
on intertidal and subtidal marine invertebrates (Vermeer  and Bourne,
1982).  Harlequins consume a wide variety of intertidal clams, snails,
small blue  mussels,  and limpets  (Koehle,  Rothe and Dirksen,  1982;
Dzinbal and Jarvis,  1982). Surf scoters and goldeneyes utilize  larger
blue mussels (Mytilussp.) obtained  by diving.   Bivalves, particularly
blue mussels (Mytilussp.) , and small clams (Macomasp.) , are well known
for their ability to concentrate pollutants at high levels (Shaw et
al,  1976) .   The crude oil  spilled from the  EVOS may  injure  marine
invertebrates that  support  sea  ducks throughout the  year  (Stekoll,
Clement, and Shaw,  1980).  Hydrocarbons may bioaccumulate in the food
chain and result in uptake of petroleum hydrocarbons by sea ducks over
a long  period (Dzinbal  and  Jarvis, 1982;  Sanger and  Jones, 1982).
This study is designed  to determine levels of petroleum  hydrocarbon
ingestion by sea ducks and document resultant physiological and life
history effects (Gay, Belisle and Patton, 1980; Hall and Coon,  1988).
A predictive model  may be constructed for harlequin duck reproductive
losses  based upon  physiological effects of petroleum contamination
resulting from the EVOS.  Pre-oil  spill baseline data are  available
on petroleum contaminant  levels in harlequin  ducks  tissue from PWS
(Irons, FWS, pers.  comm.).
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                             OBJECTIVES

A.   Develop a data base describing food habits of the six species of
     sea ducks in PWS.

B.   Obtain data  from other NRDA  studies on petroleum  hydrocarbon
     levels in marine invertebrates,  particularly blue mussels,  from
     the  PWS  area;  relate  these data  to the  levels of  petroleum
     hydrocarbons found by chemical analysis of invertebrates in gut
     samples from sea ducks  collected in oil spill and control areas;
     and  test  the hypothesis  (at  a  = 0.05)  that the incidence  of
     petroleum hydrocarbons in gut samples from  collected  sea ducks
     is higher in the oil spill areas than in the control areas.

C.   Estimate by  chemical  analysis  petroleum hydrocarbon  levels  in
     collected sea  duck tissues and body fluids within 10%  of  the
     actual value 95% of the time.

D.   Test  the hypothesis   (at  a  = 0.05)  that the  incidence  of
     petroleum hydrocarbons in tissues  of collected  sea  ducks  is
     significantly higher in 1989-91 in the oil  spill  areas  than in
     the control area.

E.   From   evidence  of  histopathology,   estimate  the   ingested
     petroleum hydrocarbon effects on morbidity and mortality of sea
     ducks.   This information may  be  related  to other studies  to
     identify  changes  in   abundance and distribution  within   the
     affected areas.

F.   Test the hypothesis that productivity of harlequin ducks in the
     oil  spill area of PWS is the  same  as productivity in  control
     areas of PWS.
                               METHODS

This study compares levels of petroleum hydrocarbons in tissues of six
species of ducks collected in four study areas.  The areas exposed to
petroleum are western PWS and southwestern Kodiak Island.  The control
sites are southeastern PWS and southeastern Alaska  (north of Juneau).
Tissues were collected for evidence of both histopathological changes
and   chemical   contamination.      Analysis   of  chemical    and
histopathological samples from these ducks continues in 1991.

Female harlequin  ducks are  secretive  and nests difficult  to  find.
Therefore,  females  will be  mist-netted  and radio-tagged  at  stream
mouths in  oiled and unoiled areas  of PWS in spring  1991  and  radio-
tracked along streams to locate nesting sites.  Clutch size, hatching
success, and brood size (a productivity index)  will be  obtained from
sample nest sites in oiled and unoiled areas.
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ANOVA (Snedecor and Cochran, 1980) will be used to test the hypothesis
that  prevalence  of  petroleum  hydrocarbons  in  gut  samples  from
collected sea  ducks  is higher  in the oil spill  areas than  in  the
control areas.

Cumulative logit loglinear models (William and Grizzle, 1972; Agresti,
1984) will be used to model the  incidence of petroleum hydrocarbons
using area collected and species as explanatory variables.  Hypotheses
concerning differences  by area in incidence of petroleum hydrocarbons
will  be  tested with a conditional likelihood  ratio  statistic  for
nested models  (Agresti, 1984).  A  Bonferroni (Snedecor and Cochran,
1980) Z-statistic (Agresti, 1984) will be used to determine the nature
of the differences among areas if the main effect is significant.

Exposure of sea ducks to hydrocarbon contaminated prey may result in
physiological effects,  such as changes in the amount of  body fat.  Sea
ducks were weighed and fat tracts  photographed.   Fat  deposition  was
classified by  condition as:  excellent,  good,  fair,  poor,  or none.
Adipose tracts  scored  were:  throat, flank,  subcutaneous,  heart  and
mesenteric.   Loglinear models (Agresti,  1984) will be  used to model
the distribution of physiological  classification  (fat  tract scores)
by area and  species.   A conditional likelihood ratio  statistic  for
nested models will be used to test the hypothesis that physiological
classification  is  independent  of  area.   If  area and  physiological
classifications  are dependent,  a Bonferroni  (Snedecor  and Cockran,
1980)  Z-statistic  (Agresti,  1984)  will  be   used   to  determine
differences among areas while controlling for physiological effect.

Tissues  were  collected for  either  chemical   analysis  (presence,
absence, or degree of petroleum residue)  or histopathology.  Results
are  being compared to unexposed  specimens  from  "clean"  (unexposed
control)  areas.   Choice of materials  and tissues,  handling,  and
discussion  of  results  are according  to published  guidelines  for
interpreting residues of petroleum hydrocarbons  in wildlife tissues
(Hall and Coon, 1988).


                            BIBLIOGRAPHY

Agresti, A.   1984.  Analysis of ordinal categorical data.  John Wiley
     & Sons, New York.   287 pp.

Dzinbal, K.A.  and  R.L. Jarvis.    1982.   Coastal  feeding  ecology of
     harlequin ducks in Prince William Sound, Alaska,  during summer.
     pp. 6 - 10 in  Marine birds: their feeding ecology and commercial
     fisheries  relationships.   Nettleship, D.A., G.A.  Sanger,  and
     P.P. Springer, eds.  Proc. Pacific Seabird Group Symp., Seattle,
     WA., 6-8 Jan. 1982.  Can. Wildl. Serv. Spec. Publ.

Hall,  R.J.,  and  N.C.   Coon.     1988.    Interpreting  residues  of
     petroleum  hydrocarbons  in  wildlife tissues.   U.S. Fish  and
     Wildl.  Serv., Biol. Rep. 88(15).  8 pp.

                                 84

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Hogan, M.E.  1980.   Seasonal habitat  use  of Port Valdez, Alaska  by
     marine birds.  Unpublished administrative report.  U.S.  Fish and
     Wildl. Serv., Anchorage, Ak.  25 pp.

Isleib, M.E. and B. Kessel.  1973.  Birds  of the  North Gulf Coast -
     Prince William Sound Region, Alaska.   Biol.  Pap.   Univ.  Alaska
     14.  149 pp.

King, J.G. and G.A. Sanger.  1979. Oil vulnerability index for marine
     oriented  birds.    pp.  227-239    in  J.C.  Bartonek  and  D.N.
     Nettleship  (eds.).   Conservation  of  marine  birds  in  northern
     North America. U.S.  Fish  and Wildl. Serv., Wildl. Res.  Rep. 11.
     Washington, D.C.

Koehl, P.S., T.C. Rothe,  and D.V. Derksen.   1982.  Winter food habits
     of Barrow's goldeneyes in  southeast Alaska,  pp. 1 - 5 in Marine
     birds:   their  feeding   ecology  and  commercial  fisheries
     relationships.    Nettleship,  D.  N.,  G.A.  Sanger,  and  P.F.
     Springer, eds. Proc. Pacific Seabird Group Symp., Seattle,  WA.,
     6-8 Jan. 1982.  Can. Wildl. Serv.  Spec. Publ.

Sanger, G.A. and R.D. Jones, Jr.  1982.  Winter feeding ecology and
     trophic relationships of  oldsquaws and white-winged scoters  on
     Kachemak Bay, Alaska,  pp. 20-28 in Marine birds:  their feeding
     ecology  and commercial fisheries  relationships.    Nettleship,
     D.N.,  G.A.  Sanger,  and  P.F.  Springer,  eds.    Proc.   Pacific
     Seabird Group Symp., Seattle, WA., 6-8 Jan.  1982.   Can.  Wildl.
     Serv. Spec. Publ.

Shaw,  D.G.,  A.J. Paul, L.M. Cheek,  and H.M.  Feder.    1976.  Macoma
     balthica: an  indicator  of oil  pollution.   Mar. Poll.  Bull.   7
     (2): 29-31.

Snedecor, G.W. and W. G. Cochran.  1980.   Statistical  methods.  Iowa
     State University Press.  Ames, Iowa.   507 pp.

Stekoll,  M.S.,   L.E.   Clement, and  D.G.   Shaw.   1980.  Sublethal
     effects of  chronic  oil exposure on the intertidal  clam Macoma
     balthica.  Mar. Biol.  57: 51-60.

Vermeer, K. and N. Bourne.   1982.  The white-winged  scoter diet in
     British Columbia:  resource partitioning with other scoters,  pp.
     30 -38  in  Marine birds:  their  feeding ecology and commercial
     fisheries relationships.   Nettleship, D.A.,  G.A.  Sanger,  and
     P.F. Springer, eds.  Proc.  Pacific Seabird Group Symp.,  Seattle,
     WA., 6-8 Jan. 1982.   Can.  Wildl. Serv. Spec. Publ.

Williams, O.D.  and J.E.   Grizzle.   1972.   Analysis of  contingency
     tables having ordered response categories. Jour. Am. Stat. Assn.
     Vol. 67: 55-63.
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                               BUDGET

Salaries                      $ 87.9
Travel                          30.0
Contracts                       40.0
Supplies                        12.0
Equipment                        9.0

Total                         $178.9
                                86

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                 FISH/SHELLFISH INJURY ASSESSMENT

The grounding of the tanker Exxon Valdez discharged crude oil into
one  of  the richest marine fisheries  communities of  the  United
States.  Although oil contamination was most severe within PWS, the
oil spread into large portions of the Gulf of Alaska (GOA),  Lower
Cook Inlet  (LCI), Shelikof  Strait,  and  other North Pacific  Ocean
waters off the coasts of Kodiak and the Alaska Peninsula.  The fish
and  shellfish populations inhabiting these  marine  and estuarine
waters form integral parts of a vast and complex ecosystem,  which
also  includes  various  other  invertebrate  species,   birds,  and
mammals  (including humans).

For example, the various life history stages of Pacific  herring are
important  forage species  for various  piscivorous fishes  (e.g.
Pacific salmon, halibut, etc.), birds (gulls, cormorants,  eagles,
loons,   etc.),   mammals   (sea   lions,   seals,   whales,   etc.),
invertebrates (crabs),  and are used  for subsistence and commercial
purposes.   Outmigrating smolts of  Pacific  salmon  are important
seasonal prey items for a  variety  of predatory  fish  and  marine
birds.  Maturing salmon in the high seas and adult  salmon returning
to  inland  waters,  are the major portion  of the diet of  marine
mammals such  as  sea lions,  seals,  and killer whales.   Salmon are
also the  summer mainstay for  eagles and many  species of  gulls.
Spawning adults in the streams constitute almost 100% of the summer
diet for bear and some river  otter  and are a very important link
between the marine and terrestrial ecosystems.  Salmon carcasses in
streams, estuaries,  and lakes are a crucial source  of nutrients for
planktonic communities and benthic organisms, which represent the
bottom rungs  of the  food chain for a wide variety of animals.

Various fish and shellfish species are also important components of
subsistence,  commercial, and sport fishery harvests.  Communities
such  as  Tatitlek,   Chenega Bay,  and  English Bay  depend  upon
subsistence fisheries  in  PWS and LCI for the  very  existence of
their  residents.    The  ex-vessel  value of  commercial fish  and
shellfish catches within PWS and other affected areas was estimated
to be $1.3 billion.   The largest recreational fisheries in Alaska
for salmon, halibut, and  rockfish center  in Homer  and Seward; a
total of 300,000 angler days was recorded from these areas in 1987.
Finally,  many non-consumptive  users of  fish and wildlife  also
utilize the waters affected by the oil spill.  Injury to fish and
shellfish  populations  and  resulting  alterations to  ecological
communities would certainly diminish the value of  the area to this
group of people.

Bioassays prior to EVOS  using crude  oil  from Prudhoe Bay and other
areas have shown  that exposure  to  concentrations  as low as a few
parts per billion in seawater  will  cause loss of  limbs in Tanner
crab, immediate death of eggs and larvae of herring, and death of
Dungeness crab and various shrimp  species.   To assess the type and
extent of injury  to marine  fish and shellfish communities by the

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EVOS, a  series  of Fish/Shellfish (F/S) studies was  developed by
investigators from various  State and Federal agencies.   Species
were selected  for study  based  on their  value  as indicators of
injury,  their role as key species within  the  ecosystem,  or their
direct importance to  man as components of subsistence, commercial,
or sport harvests.

Comparisons  of  the  abundance of  larvae,  juveniles,  or  adults
between  oiled  and  unoiled  waters  were  chosen  as  the  basic
experimental units.   In some studies,  oiled and  unoiled  waters
pertain to different  geographic areas; in other studies these terms
relate to  the same area  or populations before and after the oil
spill;  in  the  remaining studies  these terms refer  to  different
areas and populations before and after the  spill.  Contamination of
individual fish and shellfish is determined by analysis of  tissue
samples,  bile samples, or testing for induction of specific enzymes
associated  with  hydrocarbon  exposure.    Injuries  to   fish  and
shellfish populations resulting from the oil spill may be expressed
as lethal  (e.g.,  mortality to specific life history  stages)  or
sublethal  (e.g., decreased growth, reproduction potential,  etc.).
Such injuries to populations could cause losses in harvests and use
of these species by man,  and result in undesirable alterations of
natural communities.

Project  proposals were  reviewed and  modified through  comments
provided  by  State and  Federal  agency  staff  members, State and
Federal  attorneys,   various  experts retained by  the State  and
Federal governments,  and many corporate and private  individuals.
Based on these comments and results from 1989 and 1990 studies, a
number of changes  were made for the 1991 fisheries program.  Salmon
studies F/S 1,  2,  3,  4, 27,  28 and 30 were continued another year.
That portion of F/S  3 relating  to  tagging of hatchery  and wild
stock  salmon was recommended  to  be  accomplished  through  the
restoration  program  while  tag  recovery  from  adult salmon  and
analysis would  be continued within this damage assessment F/S 3
project.    Salmon  studies F/S  7 and  8 were funded  as  necessary to
conclude these projects in 1991.  Dolly Varden and cutthroat trout
study F/S  5, herring  study F/S  11  and clam study  F/S 13 were
approved for continuation in 1991.  The injury to shrimp  study F/S
15,  injury to rockfish study F/S 17 and  injury to demersal fish
study F/S  24 became  subtidal  studies ST 5,  6, and 7  respectively
and were recommended for  continuation  in  1991.   Trawl assessment
study F/S 18 was funded only as necessary to conclude this project
in 1991.   The crab study  outside  PWS  (F/S 22)  was not approved for
continuation in 1991.
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FISH/SHELLFISH STUDY NUMBER 1

Study Title:   Injury to Salmon Spawning Areas in PWS

Lead Agency:   ADF&G


                           INTRODUCTION

The recent annual production of wild stock pink salmon in PWS has
ranged from 10 to 15 million fish.  Chum salmon returns have ranged
from 0.8 to 1.5  million  fish.   Much of the spawning for pink and
chum salmon occurs in intertidal areas (up to 75% in some years).
Intertidal spawning  areas  are  susceptible to marine contaminants
and  the  March  24,   1989,   EVOS   may  adversely  affect  spawner
distribution and success  in PWS.  To detect injury to pink and chum
salmon  stocks,  intertidal  contamination  will be  documented and
correlated with  trends in  adult  returns.   Return  estimates are
based  on accurate  appraisals   of  catch  and escapements.    This
project  is  designed to document oil contamination  of intertidal
spawning  habitat;  provide  accurate  estimates  of  wild  stock
escapements; and provide estimates of intertidal and  upstream areas
available for spawning. F/S Study  3 provides  estimates of the wild
stock component of the commercial  catch.   Results  from F/S Study 3
and this study will  be combined to estimate total return of wild
stocks.  F/S  Study  2  estimates eggs and fry per square meter and
egg to fry survival  by tide  zone in a subset of the streams in this
study.  Egg and fry density  and survival data from F/S Study 2 will
be combined with spawner density data by tide zone from this study
and  historic  average  fecundity  data  to  estimate  total  egg
deposition and egg to fry survival by tide zone in  138 streams.

The ADF&G has  performed spawning ground surveys of the major salmon
spawning streams  in  PWS  since  the  late 1950's.   An aerial survey
program provides weekly estimates of fish numbers in 218 spawning
streams.  A  ground  survey  program has  provided  corresponding
estimates of fish numbers on a  subset of approximately 116 streams
during the peak of spawning.  During 1987 and 1988, funding for the
ground survey program was  severely curtailed and only 58 streams
were walked.  F/S Study 1 includes  a thorough and extensive ground
escapement  survey program  on  salmon spawning  streams  for  which
there are past ground survey data and includes additional oiled and
unoiled  streams  in western  PWS.  The study  also  includes  ground
surveys  of  salmon  streams to  document  the  presence  of oil  in
intertidal spawning habitat and the presence or absence of oil in
the tissues  of adult  salmon returns,  and  from fry outmigration
during and subsequent to the EVOS.

A total of 411 streams were surveyed in 1989 for the presence of
oil in intertidal spawning  areas and 138 streams from among the 218
in the  historic  aerial survey  program were  included  in a ground
census of pink and chum salmon escapements.  In 1990  the oil survey

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was limited to 138 streams in the escapement censusing portion of
the  project.     Mussel   samples  for  hydrocarbon  analysis  were
collected in  the intertidal portions of  the 138 streams  in the
ground censusing program in both 1989 and 1990.  The total area of
intertidal  spawning  habitat was  estimated for  each  of the 138
streams and the area of upstream spawning habitat was estimated for
100 of the 138 streams.   Total pink salmon spawning escapement at
four  streams   was  estimated  through weirs  in  1990  and  stream
residence time (stream life) estimates were made for pink salmon in
22 streams.  Tissue samples for hydrocarbon analysis were collected
from spawning adult pink salmon  in  12 oiled and 10 unoiled streams
in the ground survey program.

Based on results of the  1989  and 1990 studies, the program in 1991
will emphasize more detailed and intensive data collection on fewer
streams. Weirs will be installed on seven streams; the four streams
weired in 1990 and  three additional streams. The six streams in the
wild stock tagging portion of F/S Study  3 will be among the weired
systems  and adults  will be  sampled for  coded-wire  tags   (CWT)
applied during the 1990  field season.   Ground  surveys and stream
life  studies  will  be  continued  at  each  weired  stream  and
approximately 20 additional streams.  Oil surveys as well as mussel
and adult  salmon tissue  sampling  will  continue on all  surveyed
streams in 1991.

Results of this study will provide accurate estimates  of pink and
chum  salmon  escapement  to each  stream surveyed; will  correlate
escapement estimates  based on aerial counts  with weir and ground
counts  to estimate  past and  current   year  escapements for 218
streams included in the ADF&G aerial survey program; will provide
estimates of post oil spill distribution of spawning within stream
zones and among streams; will estimate total available intertidal
and upstream   spawning  habitat for  each  stream;  will  estimate
average stream life for pink and chum salmon in PWS; will provide
coded-wire  tag  data  for  F/S Study  3;  will  document  physical
presence  or absence  of  oil  in intertidal  salmon spawning and
rearing  habitat  and  presence or  absence of oil  in tissues of
mussels and salmon that rear or live there; and will provide an
atlas of aerial photographs and detailed maps of important spawning
sites.
                            OBJECTIVES

A.   Determine the presence or absence of oil on intertidal habitat
     used by  spawning salmon through visual observation,  aerial
     photography, and hydrocarbon analysis of tissue samples from
     intertidal mussels at stream mouths.

B.   Document the physical extent of oil  distribution on intertidal
     spawning areas.


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C.   Document the presence or absence of hydrocarbons from the EVOS
     in  the tissues  of  adult  salmon  originating  from the  fry
     outmigrations  in  1989  and  subsequent  years  in oiled  and
     unoiled areas.

D.   Estimate the  number of spawning salmon,  by  species,  within
     standardized intertidal and upstream zones for 27 streams in
     PWS.

E.   Enumerate the total intertidal and upstream escapement of pink
     and chum salmon through weirs installed on seven streams that
     are  representative  of streams  in  the  aerial and  ground
     escapement survey programs.

F.   Estimate the  accuracy  of  aerial  counts for the  218  aerial
     index streams by comparison of paired ground and aerial counts
     from the same streams on the same or adjacent  survey dates and
     by  comparison  of aerial,  ground,  and weir  counts  on  seven
     streams.

G.   Estimate average  stream  life of pink and  chum salmon  in at
     least 27 streams in PWS using a variety of techniques.

H.   Estimate pink  and  chum salmon escapements from 1961 through
     1988  for the  218  aerial  index streams  using  the  average
     observed error in the  aerial survey method  and stream life
     data from 1989, 1990, and  1991.

I.   Estimate  the  stream  area  available  for  spawning  within
     standardized intertidal and upstream zones for the 138 streams
     surveyed.

J.   Produce a catalog of aerial photographs and detailed maps of
     spawner  distribution for  the  more important  pink  and chum
     salmon streams of PWS for use in designing sampling transects
     in the egg deposition and preemergent fry studies.

K.   Enumerate adult returns to  streams  where coded-wire tags were
     applied to wild pink salmon stocks  and assist in the spawning
     ground sampling for tag recovery.

                             METHODS

This project  is  an integral part of the  study of impacts  of the
EVOS on  Pacific  salmon populations in  PWS.   Streams examined by
this  project are  a  subset  of  the anadromous  salmon  streams
monitored  by the  ongoing  ADF&G  aerial  survey  program.    Two
additional F/S studies in PWS, pink and  chum salmon  egg deposition
and preemergent  fry studies (F/S Study 2) and salmon coded-wire
tagging  studies  (F/S  Study 3) ,  will  rely on  information  about
salmon spawning and distribution data and coded-wire tag recovery
data obtained from this project.

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Streams to be surveyed will  be selected according to the following
criteria:

     1.   Stream is included in the ADF&G aerial survey program.
     2.   Stream  is included  in the  pink and  chum  salmon  egg
          deposition and pre-emergent fry project (F/S Study 2).
     3.   Stream is included in the  CWT project for wild stocks of
          pink salmon  (F/S Study 3).
     4.   Stream has been included in stream life studies conducted
          by this project in 1989 and 1990.
     5.   Stream  was   enumerated  in prior  spawning ground  foot
          survey programs.
     6.   Stream is representative of the early,  middle, and late
          run pink and chum salmon stocks in PWS.
     7.   Stream is representative of the spatial distribution of
          pink and chum salmon  stocks  in PWS  and  include streams
          from oiled and unoiled areas.

Maps of all  streams in the program prepared from aerial photographs
prior to the 1989 field season were modified and corrected during
the three survey  circuits  in 1989 and 1990 and will  be used and
updated during the 1991 field season.

A pre-season survey to mark tide zones will be conducted in June,
prior to the return of the pink and chum salmon.   The location of
tide levels 1.8, 2.4,   3.0,  and 3.7m above mean low water will be
measured from sea level using a surveyors's level and stadia rod.
Sea level at each site will be  referenced to  mean low water with
site specific, computer generated tide tables which predict tides
at five minute intervals.  Tide  zone  boundaries will be delineated
with color  coded  steel stakes.   The linear length  of  the stream
within  each intertidal zone will  be measured with  a surveyor's
chain  or  range  finder.  The linear length of the  stream in the
upstream  zone  will be measured similarly  on short  streams  and
estimated from  accurately  scaled aerial photos on  long streams.
The average stream width will be determined from systematic width
measurements taken  in  each  zone.  The number of  measurements  in
each zone will depend  on the length of the zone.  Each measurement
will be recorded  at the appropriate location on  the stream maps
prepared in 1989 and 1990.

Crews marking,  measuring,  and mapping tide zones will also conduct
foot surveys of the intertidal stream bed and adjacent beaches  to
document, map, and classify oiling. A composite sample of mussels
will be collected at  the  mouth  of  each  stream  for hydrocarbon
analyses.   Results  of  the  analyses will  be used  to document oil
impact  that the stream sustained.   Each sample  will  consist  of
enough  mussels  to provide  10  grams of  tissue  (approximately  30
mussels) for analysis.  The mussels will be collected from 0-2 m
above mean low water in the immediate vicinity of each stream mouth
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and above water to avoid contamination by hydrocarbons on the water
surface.    The samples  will  be  stored  separately  in  properly
cleaned, glass jars with teflon lined lids.

Weirs for total escapement enumeration will be installed on seven
streams in  1991.  These  seven streams include the four weired in
1990 as well as the six streams in the coded-wire tagging project
for wild  stocks  of  pink salmon (F/S  Study  3) .  The  weirs  will be
installed at or as near as possible to the 1.8 m tide  level or the
lower level of intertidal spawning.  Weir crews will record daily
passage  through  the  weir and perform  daily  ground  surveys  of
intertidal and upstream portions of the weired systems as well as
the 20 other pink and chum salmon  spawning streams.  Live and dead
pink and chum salmon will be  enumerated in standardized intertidal
and upstream zones  in each stream.  During each stream survey the
following data will be recorded:

     1.   anadromous  stream number and name (if available);
     2.   latitude and longitude of the stream mouth;
     3.   date and time  (24 hour military time);
     4.   tide stage;
     5.   observer names;
     6.   counts  of live and dead salmon by species and tide zone
          (0.0-1.8 m,  1.8-2.4 m,  2.4-3.0 m, and  3.0-3.7  m above
          mean low water and upstream); and
     7.   weather and comments on visibility,  lighting, and other
          survey  conditions.

All data will  be  recorded on pre-printed mylar data sheets which
will  overlay  a map  of the  stream.   Maps  will be  improved and
modified during the  survey to show spawner distribution within each
zone and the upstream limit of spawning.   Particular attention will
be  given  to  spawner density  and distribution observations for
streams which are also sampled during F/S Study 2.

Counts of live and dead salmon will be made for the five tide zones
(the  intertidal  zones <  1.8  m,  1.8-2.4  m, 2.4-3.0  m,  3.0-3.7 m
above mean  low water and the upstream zone) from the 1.8 m tide
level to the limit of upstream spawning on all  138  streams.    Tide
stage will be monitored continuously and survey times and direction
will be adjusted accordingly.   If the tide stage is at or below the
1.8 m level  the stream walk will begin at the mouth  of the stream
and progress upstream. The mouth or downstream  limit of the stream
will be defined as  the point where a clearly  recognizable stream
channel disappears or is submerged by salt water.  Fish seen below
the downstream limit  will  be  included as an estimate of fish off
the stream  mouth and noted   as a comment  on  the data form.  If
portions of the stream above  the  1.8 m  tide level are submerged,
the crew  will proceed  to the upstream  limit of the walk,  walk
downstream,   and  coincide the  end  of  the  walk with the  time
predicted for  the tide to be  at  or below the 1.8 m  level.   The
upstream  limit  of  a walk will be  determined  by  the presence of

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natural barriers to fish passage (i.e.  waterfalls),  by the end of
the stream, or  by  the upstream limit of spawning.   The upstream
limit of spawning will be marked on  U.S. Geological Survey color
aerial photos of each stream following each survey.

Crew members will walk together but independently count live fish
in each  intertidal  zone on moderate size  streams with  a single
channel.    Crew members  will  individually  enter their  count  on
mechanical hand tallies.   A maximum of three replicate counts may
be made for each zone at the request of either observer. Upstream
counts  in  a  single  channel  will  be  similarly  conducted  at
convenient stopping points (i.e., log jams or other clear counting
delineators).   For  large braided or branched  streams,  each crew
member will count  separate  channels or upstream forks.  To avoid
confusion with  counts  of live  fish,  counts of dead  fish will  be
recorded on the return leg of the stream walk.   Only fish that have
died since the previous count will  be tallied as dead in the daily
surveys.   To prevent  duplicate counts  between  surveys,  tails and
tags of all dead pink and chum salmon observed will be removed.  To
avoid  perpetuating  counting  biases  within  a  counting  crew,
personnel will be rotated daily.  When possible, crew members will
not be assigned to the same streams on succeeding days.

Tests for  variability among observers  and among counting crews
(observer pairs) will be conducted on all streams on a minimum of
three separate  occasions.   During each test,  all observers will
estimate numbers of live and dead pink salmon by  zone  and will
record their counts independently.   Counts  will be compared after
all test  streams have been surveyed.    Three  crews  of randomly
paired observers  will also  replicate  counts  on 10  streams and
results among observed pairs will be compared.

All streams in the  daily  foot survey  program will be included in a
stream life study.   Stream  life studies will  be modeled in part
after previous  studies in PWS  (Helle et al.  1964; McCurdy 1984).
Fish will be captured  at the stream  mouths with beach seines and
tagged with individually numbered  Peterson disk tags color coded
for day of  capture.  Tagging will be conducted at weekly intervals.
During each tagging episode  120 fish per stream will  be tagged.  If
fewer than 120  fish are available,  all  fish captured  will  be
tagged. At weired streams,  tagged  fish  will be enumerated by tag
color as they pass  through  the weir.   Live and dead fish bearing
tags will  be  enumerated  separately by color code and tag number
during daily counts of live and dead pink and chum salmon.

Stream life will be  estimated using three  methods.   The first
estimate is the mean difference between date of tag recovery from
dead fish and the tagging date. A  separate estimate will be made
for each tag lot at each stream to examine changes in stream life
through time.   The  second  estimate will  be based  on  daily and
cumulative weir counts and  daily carcass counts.   Daily weir and
carcass counts will be used to estimate total fish days. Total fish

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days will then  be  divided  by the cumulative weir count to obtain
mean stream life.   The third method will be based on the difference
in days between peak live  count and the peak dead count from the
ground surveys.

The 22  streams  where  adult salmon  tissue  samples were taken for
hydrocarbon analysis in 1990 will be sampled again in  1991.  Twenty
males and 20 females will be captured at the weir on each stream.
The fish will be iced  and  flown immediately to the Cordova ADF&G
laboratory where tissues will be excised, labeled, catalogued, and
preserved.

Changes in  numbers and  distribution  of salmon  escapements  as a
result  of  the  EVOS will  be examined  by  dividing  streams  into
categories   based   on   levels   of   hydrocarbon  contamination.
Categorical  data  analysis techniques  such as  log  linear models
using chi-square statistics will be used to compare differences in
spawning  among  streams  and tide  zones.   Count  and  spawner
distribution data  will  also  be compared  with  historical stream
survey data and related  to the level of hydrocarbon impact.
                           BIBLIOGRAPHY

Helle, J.H., R.S. Williamson, and J.E. Baily.  1964. Intertidal
     ecology and life history of pink salmon- at Olsen Creek, Prince
     William  Sound,   Alaska.    U.S.  Fish  and Wildlife  Service,
     Fisheries No. 483.  Washington D.C.

McCurdy, M.L. 1984.   Eshamy District pink salmon streamlife study,
     1984.   Alaska  Department  of Fish  and  Game, Division  of
     Commercial Fisheries.   Prince William Sound Data Report No.
     94-18. Cordova
                              BUDGET

Salaries                      $119.0
Travel                           2.0
Contractua1                    116.0
Commodities                     31.0
Equipment                       20.0

Total                         $288.0
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FISH/SHELLFISH STUDY NUMBER 2

Study Title:  Injury to Salmon Eggs and Preemergent Fry in PWS

Lead Agency: ADF&G


                         INTRODUCTION

Much of the spawning for pink and  chum  salmon  (up to 75% in some
years)  occurs  in  intertidal  areas.  Moles,  Babcock,  and  Rice
(1987)  have shown  the adverse  effects  of  oil  on pink  salmon
alevins, particularly  in  salt water.   The EVOS  in  PWS occurred
immediately prior  to  emergence of  pink  and  chum salmon  from
stream and intertidal spawning areas.   These areas may have been
severely impacted by the oil spill.

This  study,   along  with  F/S Studies   1,  3,   and  4,   support  a
comprehensive  and  integrated determination  of   injury  to  PWS
salmon  stocks.   Results  will  include documentation  of oil  in
intertidal  salmon  spawning  habitat,   pre-spill  and  post-spill
estimates of  total adult  returns  of  wild and hatchery  stocks,
wild stock spawning success, wild  stock egg to  fry survival, and
early marine  survival  of  wild and hatchery stocks.   Information
on the extent and persistence of oil in the  intertidal zone will
be supplemented by  Coastal  Habitat Study 1.   The results  of F/S
Studies  1  through 4 will  be used by   Economic Uses Study  1  to
determine the extent of injury to the PWS salmon resource.

The ADF&G has sampled pink  and chum  salmon preemergent fry since
the  1960's  in order to predict the magnitude  of  future  salmon
returns.   The  fry  sampling  program has  operated at  a  reduced
level  since 1985.   The  oil spill  has the  potential to  cause
mortality to  the  critical egg and fry life  stages, and  thus an
increased  and  more   comprehensive   fry  sampling   program  is
necessary.    This  project  is  designed  to  meet this need  by
assessing the  effect of  the oil  spill on egg and fry of wild
stock pink and chum salmon.


                            OBJECTIVES

A.   Estimate the density, by tide zone, of preemergent fry in 48
     streams,  and  eggs in 31 streams  using  numbers of live and
     dead eggs and fry.

B.   Estimate egg mortality and overwinter survival  of pink and
     chum  salmon  eggs  in  both  oiled  and  unoiled  (control)
     streams.
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C.   Document hydrocarbon contamination  in  preemergent fry using
     tissue  hydrocarbon analysis, and  eggs and  preemergent fry
     using mixed-function oxidase (MFO)  analysis.

D.   Assess  any   loss   in   adult production   from  changes  in
     overwinter survival using the  results of F/S  Studies 1,  2,
     3, and 4.
                             METHODS

There  are  approximately 900  anadromous  fish  streams  in  PWS.
Preemergent  fry   sampling   from  some  of   these   streams  has
historically provided an abundance  index  for pink salmon that is
used  to forecast  future returns.    In recent  years,   25  index
systems  considered  representative   of  pink  and  chum  salmon
producing  streams  in PWS  have  been  sampled.    Prior  to  1985,
sampling had been performed on as many as 45 streams.  This study
is  designed to compare  rates of  mortality and  abundance  among
areas  with various  levels of  oil  impacts and  with data  from
sampling prior to the oil spill.

Sampling will  consist of egg deposition surveys  performed from
late  September  to  mid-October  and  preemergent  fry  sampling
conducted  from mid-March to mid-April.   Preliminary sampling was
performed on two occasions during the spring of 1989 in an effort
to  assess  fry abundance  prior  to  and immediately  after  oil
impact.   On the  first  occasion all 25  streams in  the ongoing
ADF&G  preemergent  index  program  were  sampled  along  with  14
additional  streams.   During the second event (approximately two
weeks  after the oil spill),   14 of  the  streams were  resampled
(representing both oiled and unoiled areas), and an additional 16
streams  were  surveyed   to  assess  their potential  as  egg  and
preemergent study streams.  During  September and October of 1989
and  1990  egg  sampling  was  conducted  on 31  of these  streams.
Preemergent fry  sampling was completed on  48 streams  from mid-
March to early May in 1990.

Spring  fry sampling  in  1991 will  be  conducted on  48  streams.
These  will  include  the   25   streams   in   the  ongoing  ADF&G
preemergent  index  program plus  23  additional  streams.    The
additional streams are located in central and southwest PWS where
most the oiling occurred.   New  study streams were selected using
the following criteria:

     1.   Adult salmon returns  were expected to  be great enough
          to indicate  a  high probability of success in egg and
          fry sampling.

     2.   Egg and fry sampling had been done in past years.
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     3.   Streams with low to no oil impact, i.e., controls, were
          selected in  the  immediate vicinity of  high  oil impact
          streams to help account for possible variability in egg
          and  fry   survival  due  to   different  environmental
          conditions.

Most of the streams with suspected or obvious oil impact were not
sampled prior  to the EVOS.  The  30 streams in  low  impact areas
include 27  with  a history of  sampling;  six suspected  of having
received some  impact  including four with a  history  of sampling;
and 12 streams with  oil visibly present in  the  intertidal zone,
including five with a history of sampling.

As in  1989  and 1990,  egg sampling will be  conducted in the fall
on 31  of  the  48 streams sampled  for  preemergent fry.   Streams
included in the  fry sampling program,  but not  in the egg program
are traditional  fry  sampling streams located on  the eastern and
northern  shore  of  PWS.   These  streams  are outside  the  area
studied for oil  impact  effects.   The 13  streams in  low impact
areas  left  in the  egg sampling  program  include  four  with  a
history of sampling.   Streams suspected of having some oil impact
and streams that had visible impact are included in both  the egg
and fry sampling programs.

Sampling methods are identical for  the  preemergent fry  and egg
sampling and are modeled after procedures described by Pirtle and
McCurdy  (1977).    On  each  sample  stream,  four  zones,  three
intertidal and one above tidal  influence, will  be identified and
marked during preemergent fry sampling.  The zones are 1.8-2.4 m,
2.4-3.0 m,  3.0-3.7 m  above mean low water,  and upstream of tidal
influence.   Separate linear transects 30.5  m in  length  will be
established for egg and preemergent fry samples in each zone  (one
transect for each type dig in each zone).  The transects will run
diagonally  across the  river  with  the  downstream  end  located
against one bank and  the upstream  end  against the opposite bank.
Overlapping  of  transects  will  be  minimized  to  control  the
influence  of  fall egg  sampling  on perceived  abundance  of fry
during spring  sampling.   Fourteen 0.3 m2,  circular  digs  (56 per
stream) will be  systematically made along each  transect  using a
high pressure  hose to flush  eggs and fry from the gravel.   Eggs
and fry will be caught in a  specially designed net.

Numbers of  live  and  dead fry  by  species,  as well  as  numbers of
live and dead  eggs by species,  will be recorded  from each  0.3  m2
dig.    Additional  information  such  as date,  time,  zone,  and a
subjective estimate of  overall  percent absorption of the fry egg
sacs in the sample will  also be noted.

Preemergent pink salmon  fry  will be collected from the intertidal
channels of streams.   Fry samples will be analyzed for the
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presence  of hydrocarbons  characteristic  of  those  found  in oil
from the Exxon Valdez.

Fry sampled for  hydrocarbon analysis will be collected from the
intertidal  stream bed at a level approximately 2.5  m above mean
low water.  Samples will  be collected when  the tide  is below that
level to avoid contamination from any surface oil film.  A shovel
or clam rake will be  used  to  dislodge the  fry from  the gravel. A
stainless  steel  strainer, pre-rinsed in  dimethylchloride  and
dried, will be used to  catch fry as they are swept downstream.
Captured  fry  will  be placed  in  jars with teflon lined lids and
frozen.

Fry from  each  tide zone  will  also be collected for  MFO analysis.
These  samples  will be  selected  randomly from the  digs  in each
transect.   Fry collected  for  MFO analysis will be  preserved in
buffered formalin solution in glass jars.

Numbers  of  live and dead  preemergent  fry  and  eggs will  be
summarized  by date,  stream,   level  of hydrocarbon  impact,  and
stream zone.   A  mixed effects analysis of variance  will  be used
to test  for differences in egg to  fry mortality due  to oiling
using the  31  streams  sampled for both  eggs  and preemergent fry.
Hydrocarbon results and  degree of oiling as  visually assessed by
the mapping portion  of the  assessment  of   intertidal spawning
areas will  be used to  post-stratify streams.  Degree  of oiling
and height  in the tidal zone will be treated  as fixed effects.
Height  in  the tidal  zone  is  nested  within  stream,  a  random
effect.   Analysis  of  covariance will be  used if an  ordinal
measure of  hydrocarbon  impact can be  obtained from  the analysis
of mussel tissue collected during F/S Study No. 1.

Power  of  the  test  was  estimated for  the  analysis  of variance
using data  from the 1975 and 1976 egg and preemergent fry samples
in PWS.   These data  indicated the ability to detect an increase
of  15% in  egg  to  fry mortality   (e.g.   10%  mortality  to  25%
mortality) at  a = 0.05, 95% of the time.

Specific statistics to be estimated are:

     1.   number  of  dead  and viable  eggs per  square meter by
          salmon species, stream, and stream zone;

     2.   number of dead and  live  fry per  square  meter by salmon
          species, stream,  and stream zone; and

     3.   egg  to fry  survival  by salmon  species,   stream,  and
          stream zone.
                                99

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                          BIBLIOGRAPHY

Moles, A.,  M.M.  Babcock, and  S.D.  Rice.  1987.   Effects  of oil
     exposure  on  pink  salmon,  O.  gorbuscha,   alevins  in  a
     simulated   intertidal   environment.     Marine  Environment
     Research, 21:49-58.

Pirtle,  R.B.   and  M.L.  McCurdy.  1977.    Prince  William  Sound
     general  districts  1976  pink  and  chum  salmon  aerial  and
     ground  escapement  surveys  and  consequent brood  year  egg
     deposition  and  preemergent fry index  programs.    Alaska
     Department  of   Fish   and  Game,   Division   of   Commercial
     Fisheries, Technical Data Report 9,  Juneau.

                             BUDGET

Salaries                   $  82.0
Travel                         4.0
Contractual                  144.0
Commodities                   10.0
Equipment                     19.0

Total                      $ 259.0
                               100

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FISH/SHELLFISH STUDY NUMBER 3

Study Title:  Salmon Coded-Wire Tag Studies In PWS

Lead Agency:  ADF&G


                        INTRODUCTION

Two  questions must  be  answered  to  measure  a  loss  in  salmon
production  due  to  EVOS:     1)  which  stocks  were  exposed  to
contaminated  waters  and 2)  to what  extent did  exposure  reduce
survival and production (catch plus escapement)?   This study will
contribute to estimates of survival and production for hatchery and
wild  stocks  in  oiled  and  unoiled  areas  by  quantifying  fry
outmigration, the wild and hatchery stock components of the catch,
and the hatchery escapements.

Wild stock returns of pink salmon in PWS have ranged from 10 to 15
million fish in recent  years.  Chum salmon returns have ranged from
0.8 to 1.5 million.  Additionally,  returns of pink salmon to four
PWS hatcheries now average more than 20 million fish and hatchery
chum salmon returns in excess of 1.4 million fish are expected.

Catch and  escapement data  for wild pink salmon  in  PWS  have been
collected  since  1961.   Hatchery production became  a significant
part of  the total salmon  return  in  1985.     Consequently,  pink
salmon fry tagging was initiated at three area hatcheries in 1986
to estimate  hatchery contributions to  the 1987  catch.   Similar
estimates were made for  a fourth facility  in  1987 and  1988.   F/S
Study 3 estimated catch and  survival rates of pink salmon released
from these four  PWS hatcheries  based  on tags  applied in 1988 and
1989 and recovered in  the commercial,  cost recovery and hatchery
brood stocks  in  1989  and 1990.  Tags were  also  applied to chum,
sockeye, coho, and chinook salmon released from PWS area hatcheries
and to smolts from two wild stocks of sockeye salmon in 1989.   A
similar multi-species tagging program  was conducted again in 1990;
however, tags were also applied to  smolts from one additional wild
stock of sockeye  salmon and  fry from six wild stocks of pink salmon
including  three  from oiled  areas  and three  from unoiled  areas.
Tagging in  1991  is being transitioned from damage  assessment  to
restoration.

Pink salmon tag recoveries are expected from all four hatcheries in
1991.  Recoveries are expected for chum salmon released from Main
Bay Hatchery  in  1986,  Main Bay and Solomon  Gulch  Hatcheries  in
1987, and  Solomon Gulch in 1989.  Tagged sockeye salmon will  be
recovered  from Main  Bay Hatchery releases in  1988  and  1989, and
releases  of coho  salmon from  Wallace H.  Noeremberg  (WHN)  and
Solomon Gulch Hatcheries in 1990.
                               101

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                             OBJECTIVES

A.   Estimate catch,  escapement,  and survival rates of pink, chum,
     sockeye,  coho,  and  chinook  salmon  released  from  five
     hatcheries in PWS.  Outmigrating  smolt and returning adults
     from these facilities are potentially exposed to  oil in the
     environment.

B.   Estimate catch of the combined wild stocks of pink salmon in
     PWS and using escapement  data  from F/S  Study 1,  estimate
     differences in relative  survival rates between pre- and post-
     spill brood years.

C.   Estimate survival rates of wild pink salmon from three streams
     with contaminated estuaries and  three with  uncontaminated
     estuaries.

D.   Estimate survival rates  of wild stocks of sockeye salmon, two
     from oiled areas, one from an unoiled area.
                               METHODS

Under a separate proposed restoration project, a subsample of fry
or smolt  from all hatcheries releasing  salmon into PWS  will be
tagged with  a  coded-wire  tag  (Appendix  A) .   Wild  pink  fry and
sockeye salmon  smolt  from  both oiled and non-oiled  areas of the
Sound will also be tagged  (Appendix B) .  Tags  will  be  applied at
rates  that  insure  sufficient numbers can  be  recovered  in the
commercial fishery, hatchery cost recovery harvests, and hatchery
brood  stock   collections  to  allow  researchers  to   estimate the
contribution  of each tag  release group  by  district,   week, and
processor stratum.

Four hatcheries released 615 million pink  salmon fry  in 1990. Each
of 32 release  groups were tagged at a rate  of approximately one tag
per 580 fish  released  (1 in  580). The tag rate was  held constant
across release  groups to  prevent confusion of  differential tag
mortality with variation in survival between release  groups  (Peltz
and Geiger,  1988; Geiger and Sharr,  1989).

In 1989,  chum salmon were tagged at the rate of approximately one
tag per 60 fish  released at the Solomon Gulch Hatchery near Valdez.
Tagging of Solomon Gulch chum salmon continued at the same  level in
1990 and the WHN hatchery release of 20.6 million chum salmon fry
was also tagged at a rate of approximately one tag per 480 fish.

Wild pink salmon were tagged from six  stocks examined in F/S  Study
2  in  1990;  three  from oil  contaminated  streams and  three from
uncontaminated streams.  Inclined plane traps were used to capture
fry as they  emerged.  Trapped fry were  manually  enumerated  in  1990.
Manual enumeration  will   continue  in  1991  but electronic fry

                               102

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counters will also be tested. A portion of the daily outmigration
were anesthetized and tagged.  The anesthesia and associated trauma
required the tagged  fish to  be  held separately from the untagged
fish, until they appeared to  have recovered fully from the effects
of tagging. The extent to which the survival  and  behavior of the
tagged fish can be extrapolated to other groups of salmon will be
assessed at the  time of recovery.  Approximately  40,000  fry were
tagged for each stock at tagging rates  ranging from  1 in 4 to l in
17 fish released.

Smolt in the 2.6 million fish release  of sockeye  salmon from the
Main Bay Hatchery were tagged at a rate of 1 in 21 in 1990.

Recovery  samples  are  stratified   by  district,  processor,  and
discrete time  segments  (Cochran  1977;  Peltz and  Geiger 1988).
Fifteen percent of the pink  salmon  catch and  a minimum of 20% of
other salmon  species  catches will  be scanned for fish  with  a
missing adipose fin in each time and area specific stratum. Catch
sampling will  be  done  in   four  fish  processing  facilities  in
Cordova, one facility  in Seward, and three  facilities  in Valdez.
When feasible,  sampling will  occur at facilities in Kodiak, Kenai,
Anchorage,   and Whittier and on large floating  processors.  All
deliveries by fish  tenders to these facilities  will be monitored by
radio and  by daily contact with processing plant dispatchers to
ensure the deliveries being sampled are district specific.

In  addition to  catch  sampling  at the processing  facilities,
approximately  15%  of the fish  in  the hatchery terminal  harvest
areas will  be  scanned for  missing  adipose fins.  There  will be a
brood stock tag recovery effort  at each of  the  three hatchery
facilities where tags were initially applied.  A minimum of 50% of
the daily brood stock requirements of each facility will be scanned
for fish with missing adipose fins.

The recovery of tags  from wild  stocks  of sockeye  and pink salmon
will coincide with recoveries of hatchery stocks in the commercial
catch, terminal harvest, and brood stock sampling programs.  Tags
will  also  be recovered  in  the escapements  of each tagged wild
stock.  At  each of these streams, crews will  enumerate the daily
escapement  through a  weir.    As  escapement  passes through the
sockeye salmon weirs, a portion will be scanned daily for missing
adipose fins.  At  pink salmon weirs,  daily foot  surveys  will be
conducted to enumerate fresh  carcasses  and the surveyors will scan
them for missing adipose fins.  Carcasses enumerated each day will
be marked daily to prevent duplicate counting on subsequent days.

In  the  catch,  terminal harvest,   brood stock,  and wild stock
escapement surveys, the total number of fish scanned and the total
number of  fish with missing  adipose fins will be  recorded. The
heads will  be  removed from fish with  missing adipose  fins. Each
head will be tagged  with uniquely numbered  strap  tags. Recovered
heads will  be  assembled  and pre-processed  in the  Cordova area

                               103

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office. Heads will then be sent to the ADF&G FRED Division Coded-
Wire Tag Laboratory in Juneau for decoding and data posting.

Coded-wire tag sampling forms will be  checked by the tag lab for
accuracy and completeness.  Sampling and biological data will first
be entered onto the laboratory's database. The heads will then be
processed by removing and decoding the tags, and entering the tag
code  and the  code  assigned in  the  recovery  survey  into  the
database. Samples will  be  processed within five  working days of
receipt.

The first step in the coded-wire tag analysis will be to estimate
the harvest of salmon from each tag lot in units of adult salmon.
For hatchery stocks,  a modification of  the methods described in an
ADF&G technical report by  Clark and  Bernard (1987)  will be used.
The  specific   methods,   estimators,  and   confidence  interval
estimators  are described  in  ADF&G  technical  reports for  two
previous studies of  pink  salmon in PWS  (Peltz  and  Geiger 1988),
(Geiger  and Sharr  1989).  Additional  references  on  methods  of
tagging pink salmon in PWS can be found in Peltz  and Miller (1988) .
In the case of  wild stocks, the methods, estimators, and necessary
assumptions are described by Geiger (1988) .

The contribution of a tag lot, to a fishery stratum, is estimated
by multiplying the number  of tags  recovered  in the  structured
recovery survey, by  the inverse  of  the proportion  of the catch
sampled  (the  inverse sampling  rate)  and  the  inverse of  the
proportion of the tag  lot that was actually tagged (the  inverse tag
rate). The escapement (brood  stock)  of each tag lot is estimated
using  methods   unique to  the  particular  situation.   After  the
contribution to each fishery is  estimated  by tag lot,  marine
survival is estimated by summing the estimated harvest of the tag
lot in each  fishery,  and the estimated escapement (brood stock),
and dividing by the estimated number of fish represented by the tag
code.

Total  catches  stratified by  week,  district, and  processor were
obtained from  summaries of  fish sales  receipts  (fish tickets)
issued to each fisherman.  The total hatchery contribution to the
commercial and  hatchery  cost recovery harvest  is the  sum of the
estimates of contributions in all week,  district,  and processor
strata:

             C,   = ^  X,j  ( N, /  Sj )  p;1

where:       Ct   = catch of group  t fish,
             X,;  = number of group t tags  recovered in  ith strata,
             N;   = number of  fish  caught  in ith  strata,
             Sj   = number of  fish  sampled in ith strata,
             pt   = proportion  of group  t  tagged.
                               104

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For sampled strata, we used a variance approximation which ignores
covariance between release groups  (Geiger 1988):

            V  (Ct)   =  SjXJN/feip, )2[1 - (Nrffejp,)-1].

The average tag recovery rate  for all  processors in  a week and
district will be used to estimate hatchery contribution in catches
delivered to processors not sampled for that  district and week.
Variances associated with unsampled strata will not be  calculated.
                            BIBLIOGRAPHY

Clark,  J.E.  and  D.R.  Bernard.  1987.   A  compound multivariate
     binomial   hypergeometric   distribution  describing   coded
     microwire  tag recovery  from  commercial  salmon  catches in
     southeastern  Alaska.   Alaska  Department  of  Fish and Game,
     Division of Commercial Fisheries, Informational Leaflet  261.

Cochran, W. G. 1977.  Sampling Techniques, 3rd ed. John Wiley and
     Sons, New York, New York.

Geiger, H.J.  1988.  Parametric bootstrap confidence intervals for
     estimates of fisheries contribution in salmon marking studies.
     Proceedings  of the international  symposium and  educational
     workshop on fish-marking techniques.  University of Washington
     Press, Seattle. In press.

Geiger, H.J. and  S. Sharr.  1989.  A tag study of pink  salmon  from
     the  Solomon  Gulch Hatchery  in  the  Prince  William Sound
     fishery, 1988. Alaska  Department of Fish and Game,  Division of
     Commercial Fisheries.  In press.

Peltz,  L.  and  H.J.  Geiger.  1988.  A study  of  the  effect of
     hatcheries on the 1987 pink salmon fishery  in Prince  William
     Sound, Alaska. Alaska  Department of Fish and Game,  Division of
     Commercial Fisheries.  In press.

Peltz, L. and J.  Miller. 1988.  Performance of half-length coded-
     wire  tags  in  a  pink  salmon  hatchery  marking  program.
     Proceedings  of the international  symposium and  educational
     workshop on fish-marking techniques.  University of Washington
     Press, Seattle. In press.
                               105

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                              BUDGET

Salaries                    -$  558.0
Travel                          18.0
Contracts                      442.0
Supplies                        39.0
Equipment                       18.0

Total                       $1,075.0
                               106

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Appendix A.  Coded-wire  tagging goals  for hatchery
              releases of salmon in PWS,  1991.
Hatchery
Armin F. Koernig
Cannery Creek
Solomon Gulch
Ually Norenburg
GRAND TOTAL
Solomon Gulch
Ually Norenburg
GRAND TOTAL
Solomon Gulch
Ually Norenburg
Whit tier
Cordova
GRAND TOTAL
Main Bay
GRAND TOTAL
U. Noerenburg
Cordova
GRAND TOTAL
Species
Pink
Pink
Pink
Pink
Pink
Chum
Chum
Chum
Coho
Coho
Coho
Coho
Coho
Sockeye
Sockeye
King
King
King
Projected
Release
116,000,000
140,000,000
140,000,000
225,000,000
621,000,000
1,600,000
78,000,000
79,600,000
1,000,000
20,000
2,300,000
100,000
50,000
3,470,000
3,575,000
3,575,000
600,000
60,000
660,000
Valid
Tag
Goal
193,000
234,000
233,000
375,000
1,035,000
20,000
156,000
176,000
30,000
10,000
73,500
10,000
10,000
133,500
125,000
125,000
30,000
10,000
40,000
Total
Release
Number /Harked
Tags to Ratio Number of
Order Goal Tag Codes
218,000
261,000
252,000
422,000
1,153,000
20,000
173,000
193,000
30,000
10,000
73,500
20,000
10,000
143,500
125,000
125,000
30,000
10,000
40,000
600
600
600
600
600
80
500
450
33
2
40
10
5
26
29
29
20
6
17
16
14
10
18
58
2
4
6
2
1
2
1
1
7
8
8
1
1
2
Tag
Length
Half
Half
Half
Half
Half
Half
Half
Half
Full
Full
Full
Full
Full
Full
Full
Full
Full
Full
Full
GRAND TOTAL     All   708,305,000  1,509,500 1,654,500     470    81   Both
                                    107

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Appendix B.  Coded-wire  tagging  goals  for wild stocks
                     of  salmon in PWS,  1991.
System
                  Projected
Treatment  Species  Outmigration
                           Total
                   Valid   Release
                    Tag    /Marked Number of   Tag
                    Goal     Ratio  Tag Codes  Length
Upper Herring B.
Hayden Ck.
Loomis Ck.
Cathead Ck.
O'Brien Ck.
Totemoff Ck.
Oiled
Oiled
Oiled
Clean
Clean
Clean
Pink
Pink
Pink
Pink
Pink
Pink
210,000
360,000
210,000
150,000
300,000
720,000
40,500
40,500
40,500
40,500
40,500
40,500
5
9
5
5
7
18
3
3
3
3
3
3
Half
Half
Half
Half
Half
Half
GRAND TOTAL
  All
Pink
1,950,000    243,000
18
Half
Coghill
Eshamy
Jackpot
Clean
Oiled
Oiled
Sockeye
Sockeye
Sockeye
600,000
600,000
600,000
27,000
27,000
27,000
22
22
22
2
2
2
Half
Full
Full
GRAND TOTAL
  All    Sockeye    1,800,000    81,000
                                                     22
                                           Both
GRAND TOTAL
  All      All      3,750,000    323,000     30
                                                             24
                                           Both
                                           108

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FISH/SHELLFISH STUDY NUMBER 4

Study Title:   Early Marine Salmon Injury Assessment in PWS

Lead Agencies:  ADF&G, NMFS


                            INTRODUCTION

Recruitment to  adult salmon  populations  appears to  be strongly
affected  by mortality  during the  early  marine  period,  because
mortality at this time is typically very high  (Parker 1968; Ricker
1976;  Hartt 1980;  Bax  1983) .  During  this period,  slow-growing
individuals sustain a higher mortality because they are vulnerable
to  predators  for  a  longer  time  than fast-growing  individuals
(Parker  1971;  Healey  1982;  West  and  Larkin  1987).  In  the
laboratory, sublethal hydrocarbon exposure has been shown to cause
reduced  growth  of  juvenile salmon  (Rice  et  al.  1975;  Schwartz
1985).   Thus,  in the  wild,  sublethal hydrocarbon  exposure  is
expected  to cause  reduced growth  resulting  in  increased  size-
selective predation.

Oil  contamination may also cause  reduced  survival  by decreasing
prey populations or  disrupting migration patterns. Oil  can be toxic
to littoral and pelagic macroinvertebrates (Caldwell et al.  1977;
Gundlach et al.  1983).  Hydrocarbon exposure  can damage olfactory
lamellar surfaces (Babcock 1985)  and cause an avoidance reaction
(Rice 1973) .

During  the  past   decade,   five  salmon   hatcheries   have  been
established within PWS.  These facilities,  operated by private non-
profit corporations, produced approximately  535 million juvenile
salmon in 1989. Approximately one million of these fish were marked
with a coded-wire tag  (CWT).   Recoveries  of  these marked fish in
PWS has played a major role in our assessment  of  the impact of the
oil spill.

In 1991,  the impact  assessment will be conducted by ADF&G and NMFS.
Studies conducted by ADF&G will focus on the impact of the oil on
fry  growth,  fry migratory behavior, and  fry-to-adult survival.
Studies conducted by NMFS will focus on fry abundance, growth, and
behavior and oil  contamination in the fish and  their prey. Also, an
experiment will be conducted to determine the  effects of ingestion
of whole oil on the growth and survival of pink salmon fry.
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                            GOALS

A.   Determine  the effects  of oil  contamination on  abundance,
     distribution, growth,  feeding  habits,  and behavior  of pink
     salmon fry during their early marine residency.

B.   Describe  the apparent  effect  of oil  contamination  on  the
     migration patterns of pink salmon fry in western PWS.

C.   Quantify  hydrocarbon  contamination  in  tissues of  juvenile
     salmon collected in oiled and unoiled areas.

D.   Determine the relationship between pink salmon fry growth and
     fry-to-adult survival.

E.   Determine if  hydrocarbon contamination affected the abundance
     of primary prey species of pink salmon fry.

F.   Determine the effects of ingestion of whole oil on survival
     and growth of pink salmon fry.


PART I:  Impacts of Oil Spill on Migratory Behavior and Growth

Lead Agency:  ADF&G

Further studies are needed to determine whether oil contamination
caused  reduced  growth  and survival  of  juvenile  pink  salmon
migrating  into heavily-oiled  areas  near Armin F. Koenig (AFK)
Hatchery in PWS.    This effort will  involve  (1)   estimating  fry
growth when the fish were near the areas where they were released
and recaptured, (2)  examining the effects of other  factors that may
have caused growth differences in  oiled and  unoiled  areas,  (3)
acquiring additional measures of the level of oil exposure of fry
in  oiled and  unoiled  areas,   (4)  quantifying the  relationship
between fry growth  and  fry-to-adult  survival,  and (5)  collecting
additional data on  fry  growth  and migration  in oiled and unoiled
areas of western  PWS. F/S  Study No.  4  will focus  on pink salmon,
because evidence  of injury  to  this  species has been collected in
previous years.

Otolith microstructure analysis will be used to estimate the short-
term growth of CWT fry. The  locations where the CWT  fry were
released and recaptured are known. The  growth of CWT fry when they
were near these locations  will be estimated  by measuring otolith
growth between increments  that are formed each day  (Volk et al.
1984).   This  approach  will enable  a  relatively clear  logical
association between oil contamination,  environmental conditions,
and fry growth in specific  areas.
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An association between low fry growth and oil contamination is not
sufficient  evidence  of  injury.  Water temperature  (Martin 1966;
Kepshire 1976), prey density, and prey species composition (Ivlev
1961; Parsons and LeBrasseur 1973)  strongly affect the feeding and
growth rates of pink salmon fry.  High densities of chum salmon fry
may cause declines of  epibenthic prey populations (Healey 1979);
however, it is not clear whether  this is true for pink salmon that
feed  more  on  pelagic  zooplankton  (Cooney  et  al.  1981).  A
quantitative examination of  these  factors  is  needed to determine
whether oil exposure  caused reduced fry growth  in 1989. Theoretical
and empirical techniques will  be used to  address  this problem. A
bioenergetics model will be used to estimate the relative effects
of water temperature, prey  density, and prey species composition on
fry growth given the conditions in 1989. The feeding rate of pink
salmon  fry  is  strongly  affected  by  prey  size   (Parsons  and
LeBrasseur 1973). Additional measurements of prey size composition
will be  made  on samples of  fry  collected  from  oiled and unoiled
areas  in 1989.  Multiple  regression analysis  will  be used  to
estimate relationships between environmental conditions and CWT fry
growth. Residuals analysis and other  diagnostic tests will be used
to determine whether the growth of fry in oiled areas  was different
than expected given the environmental conditions in  1989.

The amount of mixed-function  oxidase  (MFO)  activity in fish tissues
is a measure  of hydrocarbon exposure (Kloepper-Sams and Stegeman
1989).  MFO  analyses  will be  conducted on  selected  samples  of
untagged fry  to establish the degree of oil  exposure  of fish in
oiled and unoiled areas.

The scientific literature and experience at hatcheries suggest that
pink salmon fry growth is related to  fry-to-adult survival (Parker
1968; Parker  1971;  Ricker 1976;  Hartt  1980;  Bax  1983; Nichelson
1986; Taylor et al.  1987); however,  no quantitative relationship
between these variables  exists for PWS  pink  salmon.  A regression
equation relating mean fry growth to the fry-to-adult survival of
pink salmon  from specific tag lots will be estimated using data
from the 1988  and 1989 broods. Data  from the 1990  brood will be
incorporated in the regression after  the adult return in 1992. The
regression equation will be used to estimate the survival of fish
in oiled and unoiled areas.
                            OBJECTIVES

(Letters refer to goals described above)

A-l. Estimate pink salmon fry growth in oiled  and unoiled areas of
     western PWS in 1991.

A-2. Complete an  otolith microstructure analysis  on  all  CWT fry
     collected  in  1989,  1990,  and  1991.  Use  the  analysis to
     estimate  fry  growth  during  the  two  week  time  periods

                               111

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     immediately after the fish were released and immediately prior
     to recapture.  Estimate  the 95% confidence intervals  on all
     growth estimates.

A-3. Determine the amount of MFO  activity  in  selected samples of
     fry collected in  1989,  1990,  and  1991. Use the results from
     this  analysis  in  conjunction   with data   on  beach  oil
     contamination to group samples in an analysis of variance.

A-4. Conduct an analysis of variance on fry growth during the two
     week  time period  immediately after  release  using  otolith
     growth estimates from fry collected in 1989,  1990, and 1991.
     If significant differences  (p=0.05) in fry growth are found
     among tag lots or years, a multiple comparison of means test
     will be performed.

A-5. Conduct  a repeated  measures  analysis of variance on  fry
     growth during the two week  time  period immediately prior to
     recapture using otolith growth estimates from  fry collected in
     1989, 1990, and 1991.  If significant differences  (p=0.05) in
     fry  growth  are   found  among  areas  or  years,   a  multiple
     comparison of means test will be performed.

A-6. Conduct a multiple regression analysis to  estimate  the effects
     of oil  exposure  and environmental conditions  on  fry growth
     during the  two week time period  immediately after release.
     Conduct  residuals analysis  and   other  diagnostic tests to
     determine  whether the  growth of fry  in oiled areas  was
     significantly different (p=0.05)  from  the expected value give
     the environmental conditions in 1989.

A-7. Conduct a multiple regression analysis to  estimate  the effects
     of oil  exposure  and environmental conditions  on  fry growth
     during  the  two  week  time  period  immediately  prior  to
     recapture.  Conduct  residuals analysis and other diagnostic
     tests to  determine whether the growth of fry in oiled areas
     was significantly different  (p=0.05) from the expected value
     given the environmental conditions in 1989.

A-8. Test  for differences  (p=0.05)  in prey  composition between
     oiled and unoiled areas using chi-square analysis.

A-9. Test  for differences  (p=0.05) in  stomach  content  weights
     between  oiled and  unoiled  areas  using repeated  measures
     analysis of variance.

A-lO.Use a bioenergetics model to estimate  the relative effects of
     water temperature, prey density,  and prey composition on fry
     growth in 1989.

B-l. Describe CWT fry migration patterns in western PWS in 1991.


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B-2. Qualitatively  compare CWT  fry  migration patterns  in 1989,
     1990, and 1991.

D-l. Conduct a linear regression analysis to estimate  (p=0.05) the
     relationship  between mean  fry  growth and  the fry-to-adult
     survival of  pink salmon from specific tag  lots  released in
     1989 and 1990.
                             METHODS

Field Studies:

Pink salmon  fry will be  collected using beach  and  purse seines
deployed from a  6 m long  aluminum skiff.  Sampling will begin the
first week of May and extend to the end  of June.  A 40 m long beach
seine and 70 m long purse seine will be used to  capture the fish.
Methods used to  isolate, handle, and preserve CWT fry in 1989 and
1990 will be employed again in 1991 (Raymond and  Wertheimer 1990).
Samples (n=100)  of untagged fry will be retained from sites where
CWT  fry  are recovered. These samples  will  be  preserved  in 70%
ethanol for later otolith analysis.

Coded-wire tags  will be  extracted and  interrogated as  they are
recovered in the field. This  will enable  specific tag lots to be
targeted. Methods  developed by  the ADF&G F.R.E.D.  Division Tag
Laboratory for extracting  and interrogating coded-wire tags will be
employed. Damage to the  fishes' head will be kept to a minimum when
dissecting coded-wire tags. The  remains of  the head  and the body
will be placed in a pre-weighed vial and frozen.  The vials will be
weighed later on shore when accuracies of .01 g  can be obtained.

The  following criteria  (listed  in  order of  priority)  will  be
employed in making sampling decisions in the field:

     1)   Recover a minimum of 30 tagged fish from each tag lot.
     2)   Recover  fish from  each tag  lot  in  at  least  three
          different areas  during a single sampling period. Sampling
          sites  where fry were collected  in 1989 will be receive
          priority  (Raymond et al. 1990).
     3)   Recover fish  from each  tag  lot during at  least three
          different sampling periods.

Approximately 60 tag codes will be used in 1991.  Therefore, it will
not be possible to meet each of the sampling objectives  for each of
the tag lots. To circumvent  this problem,  tag lots from the same
hatchery with similar fry  size and time  of release characteristics
will be treated  as  a group. Sampling criteria  will  initially be
applied to these groups then  to individual  lots if time permits.
Tag lots or groups  having  characteristics similar to important tag
lots in the  1989 database will receive priority (Raymond et al.
1990).

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Water temperature at 1 m depth will be measured at all sample sites
using a YSI temperature meter. Temperature measurements will also
be  made  at  stations  100  m apart  along  1 km  long  transects
perpendicular  and parallel to  shore near important  fry nursery
areas  (Raymond  et al.  1990) .  A range finder  and compass will be
used to  estimate  the position of each station. At  each station,
measurements will be made at 1 m intervals from the surface to 10
m depth using a YSI meter.  The YSI meter will be calibrated weekly
with a mercury thermometer. Mercury thermometers will be calibrated
in an ice bath at the beginning  and end of the season. Temperature
transects will be run after an extended period of calm weather and
after a storm to determine the effect of wind mixing on temperature
variability.

Samples of fry  (n=60) will  be collected  from each  tag lot at the
Wally Noerenberg and AFK hatcheries  immediately before the fry are
released  to  estimate the mean  and  variance  of fry  body weight.
These samples will be placed in 10% formalin and later weighed to
an  accuracy  of  .01  g  in  the  laboratory.   At both hatcheries,
samples of CWT  (n=30) and  untagged  fry  (n=30) will  be taken from
each of two netpens at the same time. These samples will be used to
determine if  the mean and variance of fry  body weight are different
between CWT and untagged  fry in  the same netpen. Each sample taken
from the netpens will be made  from at least three subsamples taken
at various places in the pen.

Laboratory Studies:

Otolith microstructure  analysis  will be used to estimate fry growth
during the two week time  periods  immediately after  release and
prior  to  recapture.  Thin  sections  of sagittal otoliths  will be
prepared using methods developed by Volk et. al.  (1984). A computer
image   analysis  system   will   be   used  to  measure   otolith
microstructures. The number of increments and the diameter of-the
marine zone will be measured along at least two radius lines in the
posterodorsal  quadrant  of  each  otolith.  The  mean  of  these
measurements will be used in subsequent analyses.

Measurements of prey composition and stomach content weights will
be taken  from  16  additional samples  of untagged fry collected in
oiled  and unoiled  areas  where  important CWT  fry  samples  were
obtained in 1989.  Thirty stomachs will be  examined from each sample
of untagged  fry.  Prey  items in the  following categories will be
enumerated:  large calanoid copepods (>2.5  mm),  small  calanoid
copepods  (<2.5  mm), harpacticoid  copepods,  and other.  The prey
biomass  in  each  category  will be  estimated by multiplying the
number of individuals in each category by the mean dry weight of
the individuals in that category (Raymond,  unpublished data). Fish
used for stomach analysis will be weighed to an accuracy of  .01 g
before dissection.
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Data Analysis:

Otolith increment formation and  growth  may provide a more direct
assessment of effects of environmental conditions and oil exposure
on fish somatic growth over time.  Otolith growth analysis assumes
that  otolith increment  formation  is related  to  time,  and that
otolith growth  is related to fish  growth (Campana  and Neilson
1985). Linear regression analyses will be conducted first to ensure
that increment formation is functionally related to time and that
otolith  growth  is   related  to  fish  somatic growth.  If  these
relationships are significant, differences in increment formation
and otolith growth among tag lots will be tested using an analysis
of  covariance   (Neter  et  al.  1990).  This  analysis  examines
differences in both mean response and slope among the tag lots. The
analyses will use data  from all  CWT  fry collected in 1989,  1990,
and 1991.  Data from tag  lots with similar means and slopes (p=0.05)
will  be combined and regression equations  developed  to estimate
growth of CWT fry over two week time periods.  Ninety-five percent
confidence  bands will  be calculated  for  all growth  estimates
obtained from otoliths.

Analysis of  variance will be  used to  test the hypothesis  of no
difference (p=0.05)  in fry  otolith growth between oiled and unoiled
areas.  Analyses of  fry growth  over  the  two week time  period
immediately after release will focus  on differences  among tag lots
and oil exposure. Fry released from  AFK Hatchery  in 1989 entered
oiled water while those released from other hatcheries and in other
years entered unoiled water. Repeated measures  analysis of variance
(Winer 1971) will test  differences in  growth  during the two week
time  period  immediately prior  to  recapture. Variables in  the
analysis include tag lot,  treatment  (oil,  non-oil), time period,
and year. MFO analyses  and other data  will be used to categorize
oiled and unoiled areas. Repeated measures  analysis  of variance is
necessary because fry are recovered from the same sample sites over
time.  Significant differences  in  fry  otolith   growth  will  be
examined further with a multiple  comparison  of means  test  (Zar
1974) . Growth estimates from otoliths for all  CWT fry collected in
1989, 1990, and 1991 will  be used in this analysis.

A multiple regression  analysis  will  be  performed  to  determine
effects of oil  exposure and   release  conditions  on  fry otolith
growth during the two week time period immediately after release.
Data from tag lots released in  1989, 1990, and  1991 will be used in
the  analysis.  The effects of size  of release,  size of  fry at
release,   timing  of   release,   zooplankton  abundance,   water
temperature,  and oil  exposure on  fry  growth will  be  examined.
Examination  of  residuals and   other diagnostic tests  will assess
adequacy of the fit  of the  model  and any violation of assumptions.
Fry from AFK Hatchery in 1989  were released into oiled areas while
all other fry were released into unoiled areas. Influence of data
from AFK Hatchery in 1989  on  regression parameter estimates will
also be investigated  using  dummy variables (Draper  and Smith 1981) .

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A  bioenergetics  model  (Kimmerer  et  al.  1991) will  be  used  to
evaluate the relative effects of prey density, water temperature,
and fry density on fry growth. Model parameters will be taken from
studies on  pink salmon; however,  when model parameters  are not
available for pink salmon,  parameters  from other salmonids will be
used. The effect of parameter uncertainty will be investigated by
producing  model  growth  estimates for  the  probable  range  of
parameter values.  A range of growth estimates will then be produced
for the probable range of water temperature,  prey density, and prey
composition encountered by pink salmon fry in oiled  and unoiled
areas of PWS in 1989.

A  linear  regression  analysis  (Zar  1974)  will  be conducted  to
estimate (p=0.05)  the relationship between mean fry growth and the
fry-to-adult survival of fish from specific tag lots. Data from tag
lots released in 1989,  1990, and 1991 will be used in the analysis.
The  regression  equation  will   be   used   to  examine  possible
differences  between  estimated  and predicted survival  of fry in
oiled and unoiled areas in 1989.

Prey  composition   in  the   diet  in  1989  will  be examined  using
separate chi-squared  tests on the proportion  of  stomach content
weights in each of  four prey categories. The analysis will test for
differences (p=0.05)  in the proportion of stomach content weights
in each prey category between  oiled and unoiled areas. Analysis of
covariance will be used to test for differences (p=0.05)  in stomach
content weights between oiled and unoiled areas.  Variables in the
analysis will  include treatment  (oil, non-oil)  and  time-of-day,
with fish weight as a covariate.  Stomach weight will be examined
to determine if a transformation of the data is needed.

Part II. Impact of the Oil Spill on Juvenile Pink and Chum Salmon
         and their Prey in Critical Nearshore Habitats

Lead Agency:  NMFS


                           INTRODUCTION

Preliminary results from F/S Study No.  4 have documented  effects of
the  EVOS   on  juvenile   pink  salmon,  including  exposure  and
hydrocarbon tissue burden, reduced  growth  in oiled  areas,  and
changes  in  migratory  behavior   (Cooney   1990;   Raymond  1990;
Wertheimer et al.  1990) .  The hydrocarbon profiles of juvenile pink
salmon contaminated in 1989 indicate ingestion of whole oil was the
primary route  of  contamination.   Hydrocarbons  dissolved in the
water column following  the spill  were low  or undetectable (Short
1990), and thus were  unlikely  to have  been a significant source of
contamination, while  sheen and mousse were  common  in nearshore
waters of western PWS in 1989.
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Zooplankton  and  epibenthic crustaceans  are  the primary  prey of
juvenile  pink and  chum  salmon fry  during their  initial marine
residency (Kaczynski et al. 1973; Cooney et al.  1981; Wertheimer et
al.  1990).    Oil  could  be  ingested  either  directly as  small
particles, or indirectly via contaminated prey.   Oil particles from
0.1 - 1.0 mm diameter were observed as deep as 80 m following the
wreck of the tanker Arrow in Chedabucto Bay (Forrester 1971).  In
that  spill,  Conover  (1971)  found  that  zooplankton  ingested oil
particles and estimated that  20% of the oil spilled was sedimented
to the bottom as zooplankton feces.  Epibenthic crustaceans, such
as harpacticoid copepods,  may also  bioaccumulate hydrocarbons from
contaminated sediments.

Proposed  research  for continuation  in  1991  is divided  into two
phases.   The  first  is  to  complete the analysis  of the  data
collected for  juvenile salmon  in  1989  and 1990 on  exposure and
contamination by hydrocarbons; distribution, abundance,  and habitat
utilization; size and growth; feeding habits; and prey abundance.
Results  and  conclusions  regarding  extent  and effects  of  oil
contamination to juvenile salmon are preliminary and tentative at
this time because of incomplete processing and analyses  of  1990 and
some 1989 data.  The objectives of this  phase  of the project are
essentially reiterations of the objectives previously defined for
the NMFS component of F/S Study No. 4.

The second phase will examine the effects of ingestion of whole oil
on juvenile pink salmon under controlled conditions. Most research
on the  effects on  hydrocarbon exposure  to  juvenile  salmon has
focused on exposure  to water-soluble fraction  (Rice  et al.  1975;
Rice et al.  1984) or prey  contaminated with water-soluble fraction
(Schwartz 1985).   There  is little information  on  the  effects of
whole oil exposure  to pink and chum salmon. Laboratory  data on the
toxicity of whole oil is needed to  link evidence of ingestion with
observed or  speculated effects  in  pink  salmon.   Such information
also will  be valuable in  assessing  the potential for injury to
other  fish  species  utilizing  the nearshore  habitats and  food
resources exploited  by juvenile pink salmon  during their initial
marine residency.


                           OBJECTIVES

(Letters refer to goals described above)

Section 1; Completion of 1989/1990 Analysis

A-l.  Compare   the  abundance of  juvenile pink  and chum salmon
     between oiled and non-oiled areas in 1989 and 1990.

A-2.  To compare distribution and habitat utilization by  juvenile
     salmon between 1989 and 1990.
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A-3. Compare sizes and growth rates of juvenile salmon between
     oiled and non-oiled areas in 1989 and 1990.

A-4. Quantify the feeding habits of juvenile pink and chum salmon
     in terms of  fullness,  frequency of  occurrence,  biomass,  and
     Index  of Relative  Importance,  and  to compare  oiled  and
     non-oiled areas in 1990 and between 1989 and 1990.

C-l. Compare  tissue contamination  of  juvenile pink  salmon  in
     relation to the degree of environmental contamination in the
     area of capture in 1989 and 1990.

C-2. Compare MFO  induction in juvenile  pink and chum  salmon  in
     relation to the degree of environmental contamination in the
     area of capture in 1989 and 1990.

E-l. Compare  the  abundance  of epibenthic  and   zooplankton  prey
     species of juvenile salmon between oiled and unoiled areas.

E-2. Compare the abundance of epibenthic prey species of juvenile
     salmon  in  relation to  the  degree of  contamination in
     sediments of beaches in contaminated embayments in 1990.

Section 2.  Effects of oil inaestion.

F.   Determine the effects  of oil ingestion on juvenile pink salmon
     in terms of degree of contamination (hydrocarbon tissue burden
     and MFO induction), survival, and growth (measured by weight
     gain, otolith increment, and RNA/DNA ratio).
                            METHODS

Phase 1; Completion of 1989/1990 Analysis

1. Sample Processing

Sample series that are incompletely processed include hydrocarbon
samples  of  pink  salmon  tissue,  sediments,  mussels,  and water;
otolith increment analysis from pink salmon juveniles; epibenthic
pump samples  from lightly oiled  and heavily oiled  transects in
Herring Bay; and  MFO  analysis  of  pink and chum salmon juveniles.
The  hydrocarbon  samples  have  been  released  to the analytical
laboratories through Technical Services 1, and should be complete
by spring of 1991.  The otolith samples are being processed by the
Washington Department of  Fish,  and are scheduled for completion in
February,  1991.    The  epibenthic  samples  are  contracted  for
completion by August,  1991.  An additional contract for completing
the appropriate MFO samples will be let in March, 1991.
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2. Data Analysis

The  univariate approach  to analysis  of variance  (ANOVA)  of a
repeated  measures design  (Frane  1980)  will  be used  to analyze
temperature, salinity, hydrocarbon contamination data, systematic
catch  data,  pelagic  zooplankton,  and  epibenthic  sled  and pump
collections.  The factors  in the environmental data ANOVA  are time,
oil, bay/corridor, and  location, with  location nested in oil and
bay/corridor.   Three replicate  observations  of temperature and
salinity were taken for each cell.   The same design will apply to
the hydrocarbon data.   In the  systematic catch data,  the factors
considered are time, oil, bay/corridor,  location, and habitat, with
location nested in oil and bay/corridor.

A  second  analytical  approach  to  test the   hypothesis  of  no
difference in abundance of  juvenile  pink and chum  salmon between
oiled  and unoiled  locations will  be   to  use  the  nonparametric
Wilcoxon paired-ranks test (Daniel 1978) .  Differences  in abundance
between matched cells of  the a priori  pairs of oiled and unoiled
locations will be compared;  56 such  comparisons are possible for
each species.  For pink salmon, differences in abundance will also
be tested separately in bays and corridors.

Based on Box-Cox  diagnostic plots (Dixon et al.  1988),  the biomass
of zooplankton  and  epibenthos will  be transformed prior  to the
ANOVA procedure by natural  logarithms  (In)  in order to normalize
distribution  and maximize variance  homogeneity.     For  pelagic
zooplankton, the factors considered in the ANOVA will be time, oil,
bay/corridor,  and  location,  with  location  nested  in  oil  and
bay/corridor.   For  the systematic epibenthic  sled  samples,  the
factors considered will be time,  oil, bay/corridor, location, and
habitat, with  location  nested  in oil and bay/corridor.   For the
tidal transect epibenthic  sled sampling, the factors are time, oil,
location, habitat, and  tide  level, with location nested  in oil.
The  number  of  species  or  species groups  of zooplankton  and
epibenthic  crustaceans will be  used  as  a  simple  measure  of
diversity (Pielou 1975), and also compared using ANOVA.

Abundance, percent gravid  females,  and percent  total harpacticoids
for primary prey species of juvenile  salmon  will be compared using
ANOVA for epibenthic pump samples  from heavily oiled and lightly
oiled beaches  within contaminated embayments.   Data  from each
embayment  sampled with  the  epibenthic  pump  will   be  analyzed
separately, with the transects  sampled nested within contamination
level.    When hydrocarbon  sediment  data  are  available,    the
abundance of the prey will be examined  as a  function of the amount
of oil actually found in the sediment samples.

Size and growth of juvenile  salmon will be examined by comparing
mean  sizes,   apparent  growth  rates,   and  the   weight/length
relationship between oiled and unoiled areas.   Mean sizes of pink
salmon will  be also analyzed  using  ANOVA  and the nonparametric

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Wilcoxon paired-ranks test.  The nonparametric approach will test
only the null hypothesis that there was no difference between fish
size in oiled and unoiled locations.

Apparent growth rates (change in size over time) will be calculated
for each habitat  type within  a location using the  regression of
natural logarithm weight over time. Analysis of covariance will be
used to  determine if fish can  be pooled over habitats  within a
sampling  locations.    Comparisons  between  oiled  and  unoiled
locations will  then be  made using  ANOVA,  where  sufficient data
exists.  The weight/length relationship will be used to compare the
condition of juvenile  pink  and  chum salmon  between oiled  and
unoiled areas, as recommended by Cone (1989).

For each prey category, dry weight,  dry weight  as a  percent of
total prey weight  in a stomach,  standardized dry weight  (dry weight
as a  percentage of fish dry weight) ,  numbers,  and numbers as a
percent of total numbers in a stomach will be calculated for each
fish.  Weight of stomach contents will  also be calculated as a
percent  of   total  weight  for  each  fish.    Index  of  relative
importance  (IRI, where IRI = % frequency of occurrence x (%number
+ %weight))   will  be calculated for each habitat type  by oil and
bay/corridor.   Minimum  variance  clustering of  standardized dry
weights will be used to identify associations among habitats, bays
and corridors, and oiled and unoiled areas.  Wilcoxon signed-rank
test will be used  to compare diet parameters between paired sets in
oiled and unoiled areas.

Phase 2: Oil ingestion experiment

Pink salmon  (Oncorhynchus gorbuscha)  fry  will be obtained from the
Auke Creek Hatchery after emergence.   Fry will be reared in 800 1
cylindrical tanks receiving 20 1m"1 single-pass filtered seawater.
Fry will be  grown to a mean size of approximately 0.6 g on BioDiet
starter feed then switched to 1 mm pellets.   At this time they will
be  randomly  allocated  into  three  oiled  treatments  groups,  a
dichloromethane control,  and untreated  controls,  and placed in
rectangular  (30  x  41  x 53 cm)  tanks receiving 1  1m"1 seawater.
There will  be 3 replicate  tanks per  treatment, for  a total of 18
tanks.   Initial numbers in each tank will  be 1000  fry.   Feeding
rates will be updated weekly, based on the estimated fry biomass in
each  tank.     Food  will  be  delivered  by  automatic  feeders,
supplemented  by hand  feeding.    Lighting  will  be natural,  and
temperature  will  be  ambient  seawater:   tanks  will  be located
outdoors.

A preliminary experiment will  start after the  fry begin feeding to
determine palatability of oiled food and how the oil behaves when
the food is  added to seawater.   Fry size  will be approximately 0.3
g, and the experiment will last one week.  Observations will
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include feeding behavior,  mortality,  and slick characteristics (if
any).   Contamination levels of food in the preliminary test will
be 0.1, 1, and 10% oil.

Food for the treatment groups will be contaminated with Prudhoe Bay
crude oil.  Food pellets plus oil dissolved in dichloromethane will
be placed in glass flasks, then rotovaped to remove the dichloro-
methane.   Samples  will be contaminated with 0.5,  1.0,  and 2.0%,
perhaps up to 10%  oil  by  weight,  depending on  the outcome of the
preliminary experiment.  Food for the dichloromethane controls will
be similarly treated, except no oil will be added:  other control
food will not be treated.   Food will be thawed shortly before use
as  needed  to  minimize  possible  evaporative  hydrocarbon  loss.
Contaminated food  will be analyzed  periodically for  hydrocarbon
levels.

Lethal and sublethal effects  of contamination  will be evaluated.
Mortality will be routinely monitored; dead fish will be removed at
least daily.  Sublethal effects will be measured as  growth in terms
of changes in mean  length  and weight and in terms  of otolith growth
and the ratio of ribonucleic  acid (RNA)  to deoxyribonucleic acid
(DNA).   Otolith increment widths and RNA/DNA ratios  are growth
processes that may be  more sensitive over  short time spans than
total somatic growth (Volk et al. 1984; Barron and Adelman 1984).
Formalin preserved fry  tissues will be examined histologically for
mixed  function  oxidase (MFO) induction.    Tissues examined will
include gills, anterior intestine/cecal epithelium, kidney, liver,
heart, vertebral cord,  and skeletal muscle.  Condition factor will
be calculated.

Before distribution  to the experimental  tanks,  110 fry  will be
subsampled randomly to  establish baseline characteristics at the
beginning of the experiment.  Subsequent subsamples of  110 fry will
be collected weekly from  each replicate.   To  avoid oiled food in
the hydrocarbon  analysis, 60  fry will be collected  before first
feeding in the morning to ensure that food and fecal material has
been voided from the gut.   These fry  will  be frozen immediately in
hydrocarbon  free  jars  with  Teflon  lids  for  later  hydrocarbon
analysis.   The  remaining 50 fry  will   collected in the  early
afternoon (circa 1:00 pm), narcotized in MS-222,  measured, blotted
dry, and weighed.  Twenty of these fish will be  randomly selected
for MFO analysis (n=10) or for possible histological/pathological
examination  (n=10)  and  placed  in   10%  buffered formaldehyde.
Stomachs will be excised from the other 30 fry in  the length-weight
sample, and weighed to determine fullness as a percentage of body
weight.   Fifteen  of these  fry  from  each sample will  also be
randomly selected,  white muscle will be removed from just posterior
to the dorsal fin  and  frozen  for RNA/DNA analysis, and the heads
removed  and stored  in   95%  reagent-grade ethanol  for  otolith
analysis.  Each sample will be labeled with a code identifying
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treatment, replicate, and fish number.  Samples from week 0, 1, 3,
6  will  be  processed for hydrocarbons,  otolith  increments,  and
RNA/DNA; samples from week 2, 4, 5 will be held in reserve.

Concentrations of hydrocarbons in preserved fry will be analyzed by
GC-MS and GC-FID using standard protocols established by Technical
Services 1.  Fry will be  thawed and viscera will be dissected from
the body.   Viscera and carcasses  will  be  analyzed separately for
hydrocarbon content.   MFO samples will be processed  by contract
with Woods Hole Oceanographic Institute.

Sagittal otoliths will be used for analysis of  otolith size and
increments.   Using  the  method described  by  Winter  (1985),  the
sagittal otoliths will be removed from each of the preserved pink
salmon  heads  by removing  the lower  jaw  and  gill  rakers  and
extracting the sagittal otoliths (visible through the clear wall of
the neurocranium) with no. 5 fine-tipped forceps.  The medial side
of the right otolith from each of the fish will be attached to an
acetate sheet  and imbedded  in casting  resin  (Schultz  and Taylor,
1987).  The otolith within the resin pellet will be thin-sectioned
via a diamond cut-off saw to expose the  plane containing the focus.
The thin section of the otolith will then be lapped and polished to
remove  excess  resin and  extraneous  scratches and cutting marks
(Neilson and Geen 1981; Schultz and Taylor 1987).  The section of
otolith will either  be viewed directly under  a transmitted-light
compound  microscope  or  the  image  from  the  microscope will  be
transferred to an image enhancement and analysis system for viewing
and analysis.   A standard axis between the  saltwater transition
check  and  the  edge  of  the  otolith  will  be  measured  in  the
posterodorsal  quadrant and  the number of rings  bisected by this
axis will  be  counted (Wilson and Larkin 1982; Volk et  al. 1984;
Deegan and Thompson 1987).  Incremental increase in the size of the
otolith along the standard axis, the number of increments and their
respective widths will be used as  parameters to test for treatment
effects.

The measurement of RNA and DNA will follow the methods described by
Bentle et al.  (1981) .  White muscle will be macerated with protease
and incubated at 37°  C with ethidium bromide for  1 hr.  The sample
will then be placed in a cuvette in a thermal-jacketed holder and
analyzed for fluorescence intensity in  a fluorometer.  RNAase will
then be added  to the sample,  the  sample will  be incubated for 45
minutes and then re-evaluated for fluorescence.  DNAase will then
be added to the  sample,  the sample incubated  for 30  minutes, and
again re-evaluated for fluorescence.  The fluorescence intensities
will be compared to  standard curves  for RNA  and DNA to determine
content of the nucleic acids.
                               122

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                           BIBLIOGRAPHY

Babcock, M.  M.  1985.  Morphology of olfactory  epithelium of pink
     salmon, Oncorhynchus gorbuscha, and changes following exposure
     to benzene: a scanning electron microscope study, p. 259-267,
     In J.  S. Gray and M. E.  Christiansen  (eds), Marine biology of
     polar regions and stress  on marine organisms.   John Wiley &
     Sons.

Bailey, J.E.,  B.L.  Wing,  and  C.R.  Mattson.   1975.   Zooplankton
     abundance and feeding habits of  fry  of pink salmon and chum
     salmon  in  Traitor's Cove, Alaska,  with  speculations  on the
     carrying capacity of  the area.   Fish. Bull. 73:946-961.

Barren, M. G.,  and  I. R.  Adelman.   1984.  Nucleic  acid,  protein
     content, and growth  of larval fish sublethally exposed to
     various toxicants.  Can.  J. Fish.  Aquat.  Sci.  41:   141-
     150.

Bax, N.J.  1983.  Early  marine mortality of marked juvenile chum
     salmon released  into  Hood Canal, Puget Sound, Washington, in
     1980.   Can. J. Fish.  Aquat. Sci. 40:426-435.

Bentle, L.  A., S. Dutta,  and J. Metcoff.  1981.  The sequential
     enzymatic  determination  of  DNA and RNA.   Analytical
     Biochemistry 116: 5-16.

Caldwell, R. S., E. M. Caldarone, and M. H. Mallon. 1977. Effects
     of a seawater-soluble  fraction  of Cook Inlet crude oil and its
     major aromatic components on larval  stages of the  Dungeness
     crab,  Cancer magister Dana.  p. 210-220 In D. A. Wolfe  (ed),
     Fate  and   effects  of   petroleum   hydrocarbons  in  marine
     ecosystems and organisms.  Pergamon Press, Oxford.

Campana, S.E. and J.D. Neilson.   1985.   Microstructure of fish
     otoliths.   Can. J.  Fish. Aquat. Sci.  42: 1014-1032.

Cone,  R.  S.  1989. The need to  reconsider the use  of   condition
     indices in  fisheries  science.   Trans.  Amer.  Fish.   Soc.
     118:510-514.

Conover, R.  J.  1971.   Some relations  between zooplankton and Bunker
     C oil  in Chedabucto Bay following the wreck of the tanker
     Arrow.  J. Fish. Res.  Board Canada 28: 1327-1330.

Cooney,  R.   T.  1990.   UAF  component.    NRDA  Status Report,
     Fish/Shellfish Project 4.

Cooney, R.T., D. Urquhart, and D. Barnard.   1981.  The  behavior,
     feeding biology and  growth of hatchery-released pink and chum
     salmon fry in Prince William Sound, Alaska.  Alaska  Sea  Grant
     Report 81-5. 114 pp.

                               123

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Daniel, W.  W. 1978.  Applied nonparametric  statistics.  Houghton
     Mifflin Co., Boston. 510 pp.

Deegan, L.  A.  and B. A. Thompson.   1987.   Growth  rate  and life
     history   events   of  young-of-the-year  gulf   menhaden  as
     determined from otoliths.  Trans. Amer. Fish. Soc. 116: 663-
     667.

Dixon, W.  J.,  P.  Sampson,  and P. Mundle.  1988.  One- and two-way
     analysis of variance with data screening, p 187-208, In
     W.J.  Dixon (ed),  BMDP Statistical software manual. Univ.
     Calif.  Press, Berkeley.

Draper, N.R. and H. Smith.    1981.   Applied Regression Analysis.
     2nd Ed., John Wiley and Sons, New York.

Forrester,  W.  D.  1971.  Distribution  of suspended  oil particles
     following the grounding of the tanker Arrow.  J. Mar. Res.
     29: 151-170.

Frane, J. 1980.  The  univariate  approach to  repeated  measures -
     foundation,  advantages, and caveats. BMDP Tech. Rep. 69.
     34p.

Godin, J.-G.J.  1981.  Daily  patterns  of feeding  behavior,  daily
     rations,  and diets  of  juvenile  pink  salmon   (Oncorhynchus
     gorbuscha) in two  marine bays of British  Columbia.  Can.  J.
     Fish. Aquat. Sci. 38:10-15.

Gundlach,  E. R.,  P. D. Boehm, M. Marchand, R. M. Atlas,  D. M. Ward,
     and D.  A. Wolfe.  1983.   The  fate  of Amoco Cadiz oil.  Science
     221:  122-129.

Hartt, A.C.   1980.  Juvenile salmonids in the oceanic ecosystem—
     the critical first summer, p. 25-57, In W.J. McNeil and D.C.
     Himsworth, eds., Salmonid ecosystems of the North Pacific.
     Oreg. State Univ. Press.

Healey, M.C.   1979.    Detritus and juvenile salmon production in
     the  Nanaimo Estuary:  I. Production   and  feeding  rates  of
     juvenile chum salmon (Oncorhynchus keta).  J. Fish. Res. Board
     Can.   36: 488-496.

Healey, M.  C.   1982.   Timing and  relative intensity  of size-
     selective mortality of juvenile chum salmon during early  sea
     life.  Can.  J. Fish. Aquat.  Sci. 39:952-957.

Ivlev, V.S.    1961.  Experimental ecology of the  feeding of fishes.
     New Haven, (trans, by D. Scott), Yale  University Press.
                               124

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Kaczynski,  V.  W.,  R.  J.  Feller,  and  C. Clayton.  1973.  Trophic
     analysis of juvenile pink and chum salmon in Puget Sound.
     J. Fish. Res. Board Can. 30: 1003-1008.

Kepshire,  B.M.     1976.     Bioenergetics  and  survival  of chum
     (Oncorhynchus keta) and pink  (O. gorbuscha) salmon in heated
     seawater.   Ph.D. Dissertation, Oregon State University.

Kimmerer, W., J. Grebmeier,  B.  Kelly, D.  Roseneau, A. Springer, M.
     Willette.    1991.    Conceptual model for the ecosystem of
     Kasegaluk   Lagoon,   Alaska.        Outer   Continental  Shelf
     Environmental Assessment Program Report  (in preparation).

Kloepper-Sams,  P.J.  and J.J.  Stegeman.     1989.     The temporal
     relationships  between  P450E  protein  content,   catalytic
     activity and mRNA levels in the teleost Fundulus heteroclitus
     following  treatment  with  B-naphthoflavone.    Arch. Biochem.
     and Biophys. 268: 525-535.

Martin,  J.W.     1966.   Early  sea  life  of  pink salmon.   Alaska
     Department of Fish and Game Informational Leaflet 87: 111-125.

Neilson,  J.  D.  and  G.  H.  Geen.   1981.   Method  for  preparing
     otoliths  for  microstructure  examination.   Progressive
     Fish Culturist 43(2): 90-92.

Neter,  J., W. Wasserman,  and M.H.  Kutner.   1990.   Applied Linear
     Statistical  Models:  regression,  analysis  of  variance  and
     experimental  designs,   3rd  ed.    Richard D.   Irwin,  Inc.,
     Homewood,  Illinois.

Nichelson, T.E. 1986.   Influences  of upwelling, ocean temperature,
     and  smolt  abundance  on   marine   survival  of  coho  salmon
     (Oncorhynelas kisutch)  in  the Oregon production area.  Can. J.
     Fish. Aquat. Sci. 43:527-535.

Parker, R.R.  1968.  Marine mortality schedules of pink salmon of
     the Bella  Coola  River,  central British  Columbia.   J.  Fish.
     Res. Board Can.  25:757-794.

Parker,  R.R.    1971.    Size selective  predation  among juvenile
     salmonid fishes in a British Columbia  inlet.   J.  Fish. Res.
     Board Can. 28:1503-1510.

Parsons, T.R. and R.J. LeBrasseur.    1973.    The availability of
     food to different trophic  levels in the marine food  chain.  In
     J.H.  Steele (ed.),   Marine Food Chains.   Oliver  and  Boyd,
     Edinburgh.

Pielou, E. C.  1975. Ecological diversity.  John Wiley & Sons, New
     York. 165 p.
                               125

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Raymond, J.,  A. Wertheimer,  and R.T.  Cooney.   1990.  Early marine
     salmon  injury assessment in  Prince William Sound:  draft
     preliminary status report. Alaska Department of Fish and Game,
     Anchorage, Alaska.

Rice, S.D.  1973.   Toxicity and avoidance tests with Prudhoe Bay
     oil and pink salmon fry.  p.  667-670.  In Proceedings of the
     joint conference  on prevention and  control  of  oil  spills.
     American Petroleum Institute, Washington, D. C.

Rice, S. D.,  D. A. Moles, J. F. Karinen, S. Korn, M. G. Carls, C.
     C. Brodersen, J. A. Gharrett,  and  M.  M.  Babcock. 1984.
     Effects  of petroleum  hydrocarbons on Alaskan aquatic
     organisms. NOAA Tech.  Mem. NMFS F/NWC-67. 128 p.

Rice, S. D.,  D. A. Moles, and  J.  W.  Short.   1975.   The effect of
     Prudhoe Bay crude  oil on survival and growth of eggs, alevins,
     and fry  of pink salmon, Oncorhynchus gorbuscha. p. 503-507,In
     1975 Conference on  prevention  and  control  of oil pollution.
     American Petroleum Institute, Washington, D. C.

Ricker, W.E.   1976. Review  of  the growth rate of and mortality of
     Pacific salmon in salt water, and non-catch mortality caused
     by fishing.  J.  Fish.  Res. Board Can. 33:1483-1524.

Schultz, D.  L.  and R.  S.  Taylor.   1987.   Preparation  of small
     otoliths for microscopic examination.  N.  Am. J. of Fish.
     Mgt. 7:  309-311.

Schwartz, J.  P.   1985.  Effects  of  oil-contaminated  prey on the
     feeding  and growth rate  of pink  salmon  fry,  Oncorhynchus
     gorbuscha.    p.  459-476,   In Vernberg,  F. John,  Frederick
     Thurberg,  Anthony  Calabrese,  and  Winona   Vernberg   (eds.),
     Pollution  and Physiology of  Marine Organisms.   U.  South
     Carolina Press.   Columbia, S.C.  545 pp.

Short, J. 1990.  NRDA Status Report, Air/Water 3.

Taylor,  S.  G.,  J. H.  Landingham,   D.  G. Mortensen,  and  A.  C.
     Wertheimer.   1987.   Pink salmon early life history in Auke
     Bay:  Residence,  growth,  diet  and  survival.  p.  273-318, In
     APPRISE  Annual  Report-1986.    Vol.  I:    Technical  Report.
     School of Fisheries, University of Alaska,  Juneau.

Volk, E.G., R.C.  Wissmar, C.A. Simenstad,  and D.M. Eggers. 1984.
     Relationship between otolith microstructure and the growth of
     juvenile chum salmon (Oncorhynchus  keta)  under different prey
     rations. Can. J.  Fish.  Aquat. Sci.  41:126-133.

Wertheimer,  A.  C.,  A.  G.  Celewycz, and M.  Carls.   1990.  NMFS
     component.  NRDA Status Report,  Fish/Shellfish Project 4.


                               126

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West, C.J. and P.A.  Larkin.    1987.    Evidence of size-selective
     mortality of juvenile sockeye salmon (Oncorhynchus nerka) in
     Babine Lake, Brititsh Columbia.   Can. J.  Fish.  Aquat. Sci.
     44: 712-721.

Wilson,  K.  H. and  P. A.  Larkin.    1982.   Relationship between
     thickness of  daily growth  increments in  sagittae and
     change in body weight of sockeye  salmon  (Oncorhynchus
     nerka) fry.  Can. J. Fish, and Aquat. Sci. 39: 1335-1339.

Winer, B. J. 1971.  Statistical principles in experimental design.
     McGraw-Hill, New York. 907 pp.

Winter,  Brian.   1985.   A method for  the efficient  removal of
     juvenile salmon  otoliths.   California  Fish and Game 71
     (1): 63-64.

Zar, J. H.  1974.  Biostatistical analysis.  Prentice-Hall, Inc.,
     Englewood Cliffs, NJ.
Salaries
Travel
Contractual
Supplies
Equipment

Total
   ADF&G

$  37.5
    2.1
   76.1
   16.5
    4.2

$ 136.4
  BUDGET

   NOAA

$  65.0
   10.0
   40.0
   27.0
   30.0

$ 172.0
                                        TOTAL
$ 102.5
   12.1
  116.1
   43.5
   34.2
$ 308.4
                               127

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FISH/SHELLFISH STUDY NUMBER 5

Study Title:   Injury to Dolly Varden Char and Cutthroat Trout
               In PWS

Lead Agency:   ADF&G


                        INTRODUCTION

The goal of  this  study is to compare the  survival  and growth of
populations of Dolly Varden char (char)  and cutthroat trout (trout)
differentially affected by the oil spill  in PWS.  This will be the
third  year  of this  project.    Trout  and  char  are  estuarine
anadromous  species  that  inhabit  PWS   (Morrow  1980).    Unlike
anadromous Pacific  salmon,  trout and char utilize  nearshore and
estuarine areas for  feeding.  Their marine migrations are not as
extensive as those  of Pacific  salmon  (Morrow  1980).   Some of the
most important stocks of these species inhabit  areas that have been
severely impacted by direct  contact with oil  including Green and
Montague Islands and Eshamy Bay (Mills 1988).  Since these species
commonly Iiv6 to age 8 (Morrow 1980) , the  potential exists for both
short-term and long-term effects  from exposure to  oil.   Study of
these species  is  crucial in that  they represent  finfish species
that inhabit the most oil-affected areas throughout most of their
lives.

The experimental design  for  this  program is based  upon the model
developed by Armstrong (1970, 1974, 1984) and Armstrong and Morrow
(1980) to explain the migratory behavior of anadromous  char.   This
model identifies two patterns of life history,  fish spawned in lake
systems and  fish  spawned in non-lake systems.  For  both groups,
juvenile char remain in freshwater residence in their natal stream
for up  to four years.   During  their  last spring  of freshwater
residence, they smolt to sea.   During  late summer  or early fall,
fish that were spawned in lake systems return to their natal stream
to overwinter  in  the freshwater  lake.   During the  spring,  they
again emigrate into marine  waters and annually return  to their
natal lake system  during late summer or early fall  to spawn and
overwinter.  Fish that were spawned in non-lake systems exhibit a
more complex migration.   Upon smelting, juvenile char search for a
lake system  to overwinter.    These fish then behave  in the same
manner as do fish  that originate in a lake  system except that they
return to  their natal stream  to  spawn and then return to their
selected lake system to overwinter.

The migratory  habits of  anadromous cutthroat  trout are less well
understood than those of  anadromous  char in Alaska  although it
appears that they exhibit similar migratory habits to char (Jones
1982).   Trout, however,  spawn in the spring as opposed to fall for
char.
                               128

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It is hypothesized  that  two detrimental impacts on these species
could result  from the presence of large  amounts  of crude oil in
marine waters:  (1)  reduced  survival;  and (2)  reduced growth.  To
test whether there was a measurable impact, three stocks of trout
and char  that overwinter in watersheds  that  issue  into a marine
environment which has been  directly  exposed to oil (oiled group)
and two stocks of trout and char that overwinter in watersheds that
issue in unoiled  areas (control group) were selected for study.

Significant changes  in stock abundance,  composition,  or dynamics
from the initial emigration  of stocks within the treatment group as
compared to stocks from the control group is assumed to be due to
contact with the oiled marine waters.  Evidence from  the literature
indicates that marine migrations can range up to 116  kilometers for
char (Armstrong 1974) and 80 kilometers for trout (Jones 1982).
Armstrong's  model  of  migratory  behavior  provides  the  basic
framework  for this  study.    First,  each of  the  study  streams
represents a  stock  of fish that annually homes to  that specific
overwintering  stream.  Second,  since overwinter residency occurs
entirely  in  freshwater,  fish  sampled  during  the 1989  spring
emigration had not  yet encountered oiled waters.  Given this, the
first sample from each stream (the emigration during  1989) provides
the baseline data for stocks in control and treatment.
                            OBJECTIVES

A.   Test if  there is no difference  in  annual  survival rates of
     char and cutthroat trout  between oiled and  control groups
     during 1989-91  and 1990-91  (the test will be  done  given a
     level of significance of alpha = 0.05).

B.   Test if there is no difference in annual growth  rates of char
     and cutthroat trout between  oiled and control groups during
     1989-91  and  1990-91 (the test will  be done given a level of
     significance of alpha = 0.05).


                             METHODS

Trout and char were  still  in  freshwater  residence at the time of
the spill,  and the opportunity existed to sample these fish during
their 1989 emigration prior to any potential exposure to an oiled
marine environment.  Data collected during 1989 became the baseline
for each system.   Therefore,  in  addition to  comparisons between
treatment and control,  comparisons are  also  possible  for each
stream within oiled  and control groups between subsequent years7
data and the 1989 baseline.

Each study stream consists  of  a  freshwater lake-river system that:
(1) is a tributary to  marine  waters that were  either impacted by
large  quantities of  oil  (oiled)  or  received  virtually  no oil

                               129

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(control); and (2)  contains stocks of anadromous trout and char.

A weir will  be installed and completely block each study stream
prior to the initiation  of the 1990 spring emigration.   A smolt
weir for sockeye salmon will operate at the outlet of Eshamy Lake
as part of F/S 3.  Sampling for char and trout will be conducted in
conjunction with this project.

During the spring sampling,  weirs  will  be used to count and sample
the emigration of trout and char from study streams.  Weirs will be
installed approximately 0.5 km upstream from the saltwater terminus
of the streams.  The weirs  will be operated  by  a two-person crew
from mid-April to  early-July.    Downstream  live  traps will  be
installed.

All fish  captured  in the trap will be examined for  presence or
absence of tags,  tag scars, and adipose fins.  Each fish containing
a tag from 1989, a tag  scar, or missing its adipose  fin will be
considered one recapture  event.  Recaptured fish with missing tags
will be retagged.   Fish  with no visible tag scar  and containing
their adipose fin (not tagged in 1989)  will also be tagged.  Each
fish captured will  be identified,  counted, and  measured (tip-of-
snout to fork-of-tail to the nearest mm) .  Scale  smears will be
collected from  the preferred area from all  cutthroat  trout and
placed individually on acetate slides in  coin envelopes.   Date,
species,   sex   (if  identifiable  from   external   maturation
characteristics) ,  and  length  will be recorded  for each  fish.
Recapture events will be recorded separately  for fish containing
tags and fish with missing tags.   Tag numbers will be recorded for
each recapture and  each fish tagged.

All fish found dead impinged on the weir or in the live box will be
examined for presence of tags and  adipose fins,  identified, and
measured as outlined above.   Sex and maturity will be determined by
internal  examination,  and  sagittal  otoliths will  be collected.
Date,  species,   sex,  length, maturity,  and  tag  number will  be
recorded.  Fish containing tags, tag scars  or missing adipose fins
will be recorded as recaptures.

Estimates of annual survival will be computed for each study site
through analysis of tag  returns.   If  all  emigrating  fish can be
examined for marks, the  estimates of  annual survival  (S)  can be
simply computed as:

S = m2/Ri

where:

     m2 = number of fish  recovered in year y+l
     Rj  = number of fish  tagged in year y.
                               130

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If the weir holds, the hypothesis of equal survival between oiled
and  control  sites  will  be  tested  using  contrast  within  a
multinomial analysis of variance (Woodward et  al.  1990).  Char and
trout  of  different   sizes   suffer  different  mortality  rates
(Armstrong 1974;  Sumner  1953)  so the size structure of different
populations will  be examined  and controlled  in  the  analysis if
necessary.

Jolly-Seber three-sample method  (Seber  1982)  will be used in the
event that each emigrating fish  cannot  be examined at the weirs.
Buckland's program RECAP  (1980) will  be used  to  generate  the
estimates and variances.  The  95% confidence intervals around the
survival  estimates will  be  compared to  tests  for  significant
differences between oiled and  control sites.

Annual individual  growth will  be calculated from the tag data as
the difference between length at  time of release and length at time
of recovery.   At each site, a box plot will be constructed for the
growth values, and observations more than  1.5 interquartiles away
from the box edge will be considered recording errors and not used
in the analysis.  An analysis of  variance will be  used to test for
significant differences  in  growth between fish  from control and
oiled groups.  Variation due to  differences  in years and initial
length can be controlled through the use of a block and covariate
in  the  linear  model  if necessary.   The power  to detect  a 5%
difference in the  growth rate  of fish from treatment and control
areas is estimated to be 90%.

The assumptions of analysis of variance are:

     1.  random sample,

     2.  normal distribution,  and

     3.  homogeneity of variance.

The assumption of  normality  will be tested using Kolomogorov's D
statistic.    If  the  data is  not  normally  distributed then  a
logarithmic or a rank transformation will be necessary.

The  homogeneity of  variance  assumption  will be tested with  a
Bartlett's test.    Again,  if the   assumption  is  not  valid,  a
transformation will be used.
                         BIBLIOGRAPHY

Armstrong, R.H.  1970.  Age, food, and migration of Dolly Varden
     smolts  in southeastern  Alaska.  J.  Fish.  Res.   Board Can.
     27:991-1004.
                               131

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        1974.  Migration of anadromous Dolly Varden (Salvelinus
     malma)  in southeastern  Alaska.    J.  Fish  Res.   Board Can.
     31:435-444.

	.  1984.  Migration of anadromous Dolly Varden char in
     southeastern Alaska - a manager's nightmare,  p.  559-570, In
     L. Johnson and B.L.  Burns (eds.), Biology of the Arctic char.
     Proceedings of  the  International Symposium on Arctic Char,
     Winnipeg,  Manitoba,  May,  1981.    Univ.   Manitoba  Press,
     Winnipeg.

Armstrong, R.H. and J.E.  Morrow.   1980.  The Dolly Varden char. p.
     99-104,  In E.K.  Balon (ed.), Chars: salmonid fishes  of the
     genus  Salvelinus.  Dr. W. Junk  b.v.,  Publisher.  The  Hague,
     Netherlands.

Buckland, S.T.  1980.  A modified analysis of the Jolly-Seber
     capture-recapture model.  Biometrics 36: 419-435.

Clutter, R. and L.  Whitesel.   1956.  Collection and interpretation
     of  sockeye salmon  scales.   International Pacific  Salmon
     Fisheries Commission, Bulletin 9.  159 pp.

Jones, D.E.   1982.   Development of techniques for enhancement and
     management of cutthroat  trout  in  southeast Alaska.  Alaska
     Department of Fish  and  Game.  Annual  Report  of  Progress,
     Project AFS-42, 23(AFS-42-10-B): np.

Mills, M.J.   1988.   Alaska statewide sport fisheries harvest
     report.   Alaska Department  of  Fish and Game, Fishery Data
     Series No. 2.   142 pp.

Morrow, J. E.  1980.  The freshwater fishes of Alaska.  Alaska
     Northwest Publishing Company, Anchorage, Alaska.   248 pp.

Seber, G. A. F. 1982.  Estimation of animal abundance and related
     parameters. 2nd edition, Griffin & Company, London. 655 pp.

Sumner, F.H.  1953.  Migrations of salmonids in Sand Creek, Oregon.
     Trans. Am. Fish. Soc. 82:  139-150.

Woodward, J.A., D.G. Bonett,  and M.L.  Brecht.  1990.  Introduction
     to  linear models and experimental  design.  Harcourt Brace
     Jovanovich Inc., San Diego,  California.  62 pp.
                               132

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                              BUDGET

Salaries                   $  230.9
Travel                         10.4
Contracts                      55.8
Supplies                       28.0
Equipment                  	0.0

Total                      $  325.1
                               133

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FISH/SHELLFISH STUDY NUMBER 11

Study Title:  Injury to PWS Herring

Lead Agency:  ADF&G


                           INTRODUCTION

The oil spill in PWS coincided with the annual migration of Pacific
herring (Clupea harengus pallasi)  to near-shore spawning areas. In
1989, a significant portion of the spawning area in PWS was located
within  areas contaminated by  oil.  Additionally, adult  spawning
herring and newly hatched juveniles traversed areas impacted by oil
and beach cleaning activities.

It was hypothesized that the oil spill  would adversely impact adult
fish through direct mortality, food shortages,  slowed growth, and
a possible reduction in fecundity. In addition,  herring eggs have
been   shown   to   be  particularly  susceptible  to  hydrocarbon
contamination due to the affinity of hydrocarbon compounds for yolk
sac material.  Impacts on egg mortality, egg hatching success, and
percent viable hatch have the  capacity to reduce the abundance and
availability of herring.  Adult and juvenile herring, as  well as
herring eggs, often form  an important item in the  diet  of marine
fishes  (e.g. salmon and halibut),  mammals  (e.g.  sea lions, seals,
and whales), and birds  (e.g.  cormorants,  ducks,  puffins,  gulls).
Herring also support  an important commercial  fishery within PWS,
worth  approximately  12  million  dollars  in 1988  and 9  million
dollars in 1990.

The goal of this project is to determine  whether the  EVOS will have
a measurable impact on populations of  herring in PWS. Accurate and
precise estimates of population abundance, age structure,  weight,
and length composition data are needed to accomplish this goal. In
addition,  the  direct  effects  of  oil contamination  on  spawning
success and egg and larval survival will be determined.

                            OBJECTIVES

A.   Expand the normal sampling of the herring population in PWS to
     increase the  precision of herring abundance, age composition,
     weight, sex ratio,  and fecundity estimates.  Specifically we
     intend to:

     1.   Estimate the biomass of  the  spawning stock of herring in
          PWS during 1991 such that the estimate is within  ± 25% of
          the true value 95% of the time.
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     2.   Estimate  the  age,  weight,  length,  and  sex  (AWLS)
          composition of herring in PWS during 1991 such that age
          composition estimates  are within  ±  10% of  their  true
          values 95% of the time.

B.   Document the occurrence of herring spawn in oiled and unoiled
     areas,  validating  the  sites  with  quantified  oil  level
     information   obtained  from   shoreline  survey  maps   and
     hydrocarbon analyses of 1989,  1990, and  1991 herring eggs and
     mussel tissue.

C.   Estimate  hydrocarbon  contamination  of,  and  physiological
     impacts on, adult herring by analyzing tissue samples.

     Test the hypothesis that the level of hydrocarbons in herring
     tissues is not related  to the level  of  oil contamination of
     the area from which the herring were sampled. The experiment
     is designed to detect a  difference of 1.6 standard deviations
     in hydrocarbon content with the probability of making a type
     I and type II error of 0.05 and 0.1,  respectively.

D.   Estimate the presence and type of damage to tissues and vital
     organs of herring sampled from oiled and unoiled areas.

     Test the hypothesis that the level of hydrocarbons in herring
     eggs is not related to the level of oil contamination of the
     area from which the  herring were sampled.  The experiment is
     designed to detect a difference of 1.6 standard deviations in
     hydrocarbon content with  the  probability  of  making  a type I
     and type II error of 0.05 and 0.1, respectively.

E.   Estimate the proportion of dead herring  eggs  in oiled and un-
     oiled areas from a subsample of study  sites that were utilized
     in the 1989 and 1990 egg mortalities study, expanding the data
     base and providing sample sites for sample collection of live
     and preserved eggs.

     Test the hypothesis that the proportion of dead herring eggs
     is not related to the level of oil contamination of the area
     from which the herring were sampled.

F.   Estimate the  hatching success, viable  hatch,  occurrence of
     abnormal larvae,  and collect embryonic and larval tissue for
     sublethal testing including cytogenetics,  MFO analysis,  and
     histopathological analyses  by  collecting herring  eggs  and
     rearing them in field and under laboratory observation.

     Test the hypothesis that hatching success, viable hatch,  and
     occurrence of abnormal larvae  are not related to the level of
     oil contamination  of the area  from  which the  herring  were
     sampled.
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G.   Estimate the  number (proportion) of  eggs removed  from the
     spawning areas (due to wave action or predation)  between the
     time of egg deposition (spawning) and the time of hatching.


                             METHODS

This project will be conducted  in  three  parts:  (1)  herring spawn
deposition estimation,  (2) herring age, weight,  length, growth, and
fecundity estimation,  and  (3)  herring egg survival and  egg loss
estimation.

Herring Spawn Deposition Estimation

The management  of  the  PWS  herring stock  is  based on a harvest
policy established by the Alaska Board of  Fisheries which specifies
a maximum  20% exploitation  rate for the combined  harvest of all
herring  fisheries.  The  allowable  harvest is  based  on  biomass
estimates established  the previous  year  modified  by the  expected
growth and survival over the year. While aerial surveys were used
to estimate  biomass  from 1973-87,  spawn deposition  surveys were
performed in 1983 (Jackson and Randall 1983) and 1984 (Jackson and
Randall 1984) , and were the primary biomass  estimate  starting in
1988 (Biggs and  Funk 1988). Aerial  surveys are easier to perform
than spawn deposition surveys, but aerial survey biomass estimates
are not as reliable because of  the varying visibility of herring
schools from  the air  and the  unknown residence time  of herring
schools  on  the spawning  grounds.   In  addition,  estimates  of
precision are not available for aerial survey biomass estimates.
The ADF&G continues to  conduct an annual  aerial survey of spawning
biomass  to  provide in season  indicators  of  run  timing  and
distribution of  spawning  activity.  This information  is  used for
planning the spawn deposition survey.

This project  represents an augmented program to assess  the PWS
herring stock's  response to the EVOS. The original  goal of the 1989
herring  spawn deposition  survey  was to  estimate the  spawning
biomass with a precision such that  the biomass estimate  would be
within ± 25%  of  the true biomass estimate 95%  of  the time under
optimal survey conditions. Fishery  managers  determined that this
level of precision was acceptable for estimating exploitation rates
and forecasting  future abundance.  If weather or  other  logistic
problems hampered  the   spawn deposition survey sampling effort,
fishery managers were  willing to tolerate  reduced precision. The
EVOS introduced  a potentially new and unknown  level of mortality on
herring stocks.   The accuracy and precision of estimates  of stock
abundance need to be assured from both oiled and unoiled areas (as
reflected  in  objectives Al  and  A2) .  The opportunity  to  estimate
herring biomass  with spawn deposition surveys  is  only available
during a relatively narrow two  week window.  After the oil spill,
the number of divers involved in the survey was increased to assure
that even  if weather problems  restricted  the available  sampling

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time, sufficient numbers of transects could still be performed. The
number  of transects  was also  increased to provide  a  level  of
precision such that the biomass estimate would be within ± 25% of
the true biomass 95% of the time.

The aerial survey project will provide a map indicating the general
location  of herring  spawning  areas.  Transects  will be  placed
perpendicular to the shoreline at locations  selected randomly from
the shoreline maps  of  spawning areas.  Divers will swim along the
transects  and  systematically  place   0.1  m2  quadrants  at  5  m
intervals. Divers will  estimate  the total  number  of eggs in each
quadrant. All  egg-containing vegetation will  be removed  from a
subset of the quadrants for later enumeration of the  number of eggs
in a laboratory procedure. These enumerated egg  counts will be used
to  correct  bias  in diver-estimated egg counts and  estimate the
precision of the  diver  estimates. The survey design is described in
detail by Biggs and Funk (1988),  and follows  closely the two-stage
sampling design of  similar surveys in British Columbia (Schwiegert
et al.  1985),  and  in  southeast Alaska (Blankenbeckler and Larson
1982, 1987). The surveys use random sampling  at  the  first stage
(transects), and systematic sampling at the second  stage (quadrants
within  transects).  Random sampling  in the  second  stage  is not
feasible because of underwater logistical constraints (Schwiegert
et al.  1985).  In addition  to the two-stage design,  the survey is
stratified by  five  areas within PWS  (southeast, northeast, North
Shore, Naked Island, and Montague Island) because of the geographic
separation of these areas  and  the potential for herring in these
areas to be discrete stocks.

Mean egg densities  along each transect will be combined to estimate
an overall average  egg  density. The observed widths of the spawning
bed  along each  of  the  transects  will be  used to estimate the
average spawning bed width. The average width, average density, and
total spawning bed  shoreline  length will be used to estimate the
total  number  of eggs  deposited  in   each  of   five area  strata
established within PWS. Using the average fecundity and sex ratio
derived from the AWLS sampling portion of this project, the total
number of eggs deposited will be converted into population numbers
and biomass. Based  on  the  variances obtained during the 1989 and
1990  surveys,  160   transects  will be  needed to  insure  that the
estimated biomass would have a 95% chance  of being within 25% of
the true biomass.   (161 and  160 transects  were conducted in 1989
and 1990 and the resulting biomass estimates had a 95% chance of
being within 19% and 23%) .

Sampling Procedure:

The general  locations  of spawning activity  will  be derived from
visible milt observed in the  water column during scheduled aerial
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surveys. This information will be compiled and summarized on maps
showing spawning locations and the number of days on which milt was
observed.

Using this information,  skiff surveys will be conducted in season,
by  members of  the  spawn deposition  dive  team,  to verify  the
accuracy of spawning area maps derived from aerial  survey data.
Diving where herring have spawned is not recommended for at least
5  days  after  spawning   activity  has  ceased   because  of  water
visibility problems caused by milt and because large numbers of sea
lions are usually present.

The shoreline  area containing herring spawn on the map, verified by
skiff survey,  will be divided into the smallest segments resolvable
on the scale of  the map (0.1 mile). A total of 160 of the shoreline
segments will be proportionally allocated to each of  five major
areas  (southeast,  northeast,   North   Shore,  Naked  Island,  and
Montague Island) based upon  the number of miles of  spawn in each
major  area.  For  example,   if  the   northeast  area  contains
approximately 25% of the  spawn in all five areas, then 25% or 40 of
the 160 transects will be placed in the northeast area. Transects
will  be selected  at  random from  all  of  the spawn-containing
shoreline segments within each area. Each transect will be assigned
a number and its location drawn on waterproof field maps that can
be taken out in  the dive skiff. The dive team leader will determine
the exact transect location within the  randomly  selected shoreline
segment by identifying a  shoreline feature (tree,  rock,  cliff,
etc.) located  above the high tide line as the dive skiff approaches
the  shore, but before  bottom  profiles,  bottom vegetation,  or
herring spawn are visible from the skiff.

A  0.1  m2 quadrant  constructed of PVC  pipe  will be  used  for  the
sampling frame.  A depth gauge and compass will be fastened to the
quadrant. Data will be recorded on pre-printed single matte mylar
forms  attached to   PVC  clipboards,   using  a  large  weighted
carpenter's pencil attached  to the clipboard.  Normally  the dive
team leader will make egg density estimates and record data while
the assistant diver sets and follows the compass course, measures
distances, and  carries and places the quadrant.

Sampling along  the transects will occur in the following manner:

     1.   A compass  course perpendicular to the shoreline at the
          transect location will be set on the compass attached to
          the sampling quadrant.

     2.   The first quadrant will be placed within the first 5 m of
          spawn by tossing the quadrant.

     3.   The lead diver will estimate and record  the number of
          eggs  in the quadrant.  The  number of  eggs is normally


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          recorded  in units of  thousands.  The  vegetation type,
          percent cover, substrate, and depth are also recorded.

     4.   The assistant diver will measure four complete 1 m hand-
          spans offshore, along the compass course. Halfway through
          the fifth hand-span, the assistant diver will gently toss
          the quadrant ahead approximately one-half meter and allow
          it to  come  to rest.  The lead diver then  makes another
          estimate at the new quadrant location.

     5.   This process continues every 5 m until the apparent end
          of the spawn is found.  Divers will verify the end of the
          spawn by swimming at least  an additional  20  m  past the
          end of the spawn,  unless a steep  drop-off is encountered.

Data codes have been developed for the vegetation  types and species
that are  encountered  in PWS.  If more than one is present  in the
quadrant  sampled, the  three most common are  recorded on  the data
forms. Percent  cover  is a  simple estimate  of the  percentage of
plant cover that exists within the quadrant sampled  (e.g., if half
the area  is covered, the cover is 50%).

Approximately every fifth quadrant will be used as a special diver
calibration sample.  Both divers will estimate the  number of eggs in
the quadrant  in  a manner such that neither can see  the other's
estimate.  Divers  will  attempt  to  remove  all  egg-containing
vegetation and scrape  eggs off rock substrate, placing the material
in numbered mesh  bags.  A sample size goal of 80 calibration samples
per diver was established,  including 20 in each of four vegetation
categories (eelgrass,  fucus, large brown kelp, hair kelp), based on
1988,  1989,  and  1990  survey results. Calibration  samples  should
also  be  spread  over  a wide  range of egg  densities. The spawn
deposition  project  leader  will  track   the  number  of  samples
collected by each diver by  vegetation group and density to ensure
that sufficient  calibration samples are  taken in  each category.
Upon completing a dive shift, calibration sample material will be
removed from the numbered mesh bags  and placed  in Nalgene Ziploc
bags. Gilson's solution will be poured over the sample so that all
material  is  completely immersed. A  label will  be made  for each
sample  (preferably  in pencil on mylar)   containing  the  transect
number, both diver's estimates, date,  and  vegetation type. Five or
6 calibration  sample  bags  can  be stored in  a  5 gallon plastic
bucket. Samples should not  be stacked over one another to prevent
spilling and mixing. Procedures for the enumeration of the number
of eggs  in  each  calibration sample are described,  including the
formulas used to prepare Gilson's solution and the other chemicals
used for sample processing.
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Data Analysis:

Biomass Estimation

The 1991 spawn deposition  survey  will  conform with the 1988-1990
spawn deposition surveys in PWS (Biggs  and Funk 1988). The overall
biomass estimator is:
         (T •  B')
     B = 	,                                              (1)
          (1 ~ R)

where,

     B  = estimated spawning biomass in tonnes,
     T  = estimated total number of eggs (billions) deposited in an
          area,
     B' = estimated tonnes of  spawning  biomass required to produce
          one billion eggs, and
     R  = estimated proportion of  eggs  disappearing from the study
          area from the time of spawning to the time of the survey
          (egg loss).

The estimates  for T  and B'  are  derived from  separate sampling
programs and are  thus independent.  Ignoring the unknown variability
in R,  the estimated variance  for the  product  of the independent
random variables T and B7, conditioned on R is:

                 [T2Var(B')  + B'2Var(T) - Var (T)-Var(B')]
     Var(BJR)  = 	,      (2)
                                  (1-R)2

where,

     Var(B') = an unbiased estimate of the variance of B', and
     Var(T)   = an unbiased estimate of the variance of T
               (Goodman 1960).

The total number of eggs deposited in an area is estimated from a
two-stage sampling  program with  random sampling at  the primary
stage,  followed  by systematic  sampling at the  secondary stage,
using a sampling design similar to that described by Schwiegert et
al. (1985).  In computing variances based on the  systematic second
stage samples it is assumed that  eggs are randomly distributed in
spawning beds with respect to  the 0.1 m2  sampling unit. While this
assumption was not  examined,  in  practice the  variance component
contributed by the second sampling stage was much smaller than that
contributed by the first stage, so that violations of this
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assumption would have  little effect on the overall variance.  The
total number of eggs (T), in billions, in an area is estimated as:

     T = N •  y •  icr6 ,                                         (3)

where,

     N      = L/v'o.l =  the total number  of  possible transects,
     L      = the shoreline length of the spawn-containing stratum
              in meters,
     Vo.l   = 0.3162 m  =  width of  transect strip,
     y      = average estimated total number  of  eggs  (thousands)
              per transect,  and
     10"6    = conversion from thousands to  billions of eggs.

The average total number of eggs per transect strip (in thousands)
is estimated as the mean of the total eggs (in thousands)  for each
transect strip using:

              Z  y;
          Y= —	,                                           (4)
                 n

where,
     Yi = average quadrant egg count in transect i (in thousands of
          eggs) ,
     i = transect  number,
     MI = Wj/v/0.1 = number of possible quadrants in transect i,
     W; = transect  length in meters,  and
     n = number of transects actually sampled.

The average quadrant  egg count within ,a transect, ~y;,  is  computed
as:
          y, - -^ - f                                         (5)
                  nij

where ,

     j   = quadrant number within transect i,
     m;   = number of quadrants actually sampled in transect i,  and
     Yij   = adjusted diver-estimated egg count  (in  thousands of
           eggs)  from the diver calibration  model  for quadrant j
           in transect i.

The variance of  T is similar to that given  by  Cochran (1963)  for
three stage sampling with primary units of equal size, although in


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this  case  the expression  is modified because  the primary  units
(transects)  do  not  contain equal  numbers  of  secondary  units
(quadrants), and the variance term for the third  stage comes from
the general linear model used in the diver calibration samples:


Var(T) = N2(10^)2[	— •  Sl2 + ^—^	  • s22 + —^-2-  s32],   (6)
                  n              Snij              Sitij


where,
           n         .
           2 (^. - ^)
     Sj2 = —	 = variance among transects,
               n-1


          n   , "Ji  *V'J ~ Yi)
     s2  = S Mj s 	 = variance among quadrants,


      ,   n ni;
     s3z = S s Var(yjj) =  sum of  the variances of the
          1=13~1             individual predicted quadrant egg counts
                           from the diver calibration  model,
           n
     fl   -  	 =  proportion of possible transects sampled, and
           N
     f2  = 	 = proportion  of  quadrants  sampled within transects
           Mj    (same for all transects)  .

Diver Calibration:

Diver observations  of vegetation species will be aggregated into
four vegetation  categories based  on structural  and  phylogenetic
similarities of plants in the quadrant: eelgrass, fucus, hair kelp,
and  large  brown  kelp.  Diver estimates   of   egg  numbers  are
approximately proportional to  laboratory-enumerated counts,  but
systematic biases in  the diver estimates can be  accounted for by
vegetation type and density (Biggs  and Funk 1988). Individual diver
effects  were not significant  in  the  1988  and  1989 survey,  but
potential differences among individual divers will be examined. The
basic  form  of  models  used  to  account  for  biases  in  diver
observations is:

                    a   Dj   Vk     Bjk   e
          Yijk  =  e •  e   • e  •  Xijk  •  e   ,                       (7)
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where,

     a  = a constant,
     Dj  = parameters representing the effect of j* diver,
     Vk  = parameters representing the effect of the k*
          vegetation type,
     Bjk = parameters controlling the functional  form of the
           relationship between the diver estimate and laboratory-
           enumerated egg count  for diver j in vegetation type k,
     Yjjk = the 1th laboratory egg  count in the vegetation-diver
          stratum jk,
     Xijk = the i* diver estimate  in vegetation-diver stratum jk,
          and
     e  = a normally distributed random variable with mean 0 and
          variance a2.

A multiplicative-effect model is  chosen because relative estimation
errors are expected to change with egg density. The distribution of
laboratory-enumerated egg  counts for a  given diver estimate was
positively skewed  in the  1988  and 1989  surveys  (Biggs  and Funk
1988, Biggs in press),  so that the  logarithmic transformation used
to estimate the parameters of the multiplicative-effect model also
stabilized the  variance  and corrected  the  skewness of  the egg
density estimates.  After  a  logarithmic  transformation,  model 7
becomes:

             l°ge(Yijk)  =  a + Dj + Vk + jyioge(Xijk)  +  e         (8)

where, 6jk = the  slope of the  relationship between  the logarithm of
the diver estimate and the logarithm of the laboratory-enumerated
egg count.

In  logarithmic  form, the  model comprises  a linear  analysis of
covariance problem with two factor effects (vegetation and diver)
and one covariate  (diver-estimated egg number). The SAS Institute
Inc.  (1987) procedure  for general linear models  will  be used to
obtain least squares estimates of parameters and evaluate variance
components.  In  addition  to  the   two   factor  effects  and  one
covariate, terms for diver-vegetation group interactions, density-
vegetation group interactions and density-diver interactions will
be considered in the analysis of covariance. Three-way and higher
level  interaction  effects  will not be  considered because the
objective  is  to derive  a simple  model  with a  relatively small
number of parameters. Backward stepwise procedures will be used to
determine  subsets  of the  six effects  that explain .the maximum
amount of  variability  in  the data  with the smallest  number of
parameters. During the backward  stepwise procedures, effects will
be included or eliminated from the model based on the probability
level of F ratios for partial sums of squares.
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Translation  of  the predicted values  from the logarithmic  model,
equation (8), back to the original scale,  equation (7),  requires a
correction  for  bias.  The bias  in the expected value  of  Yijk  is
exp(%a2) when the true variance of Yijk, a2,  is known.  Laurent  (1963)
gives an exact expression for the bias correction that incorporates
additional terms when a2  is estimated  from a sample.  For the diver
calibration  data,  the biases  in  estimating a2 from  a sample were
less than 0.05%  (Biggs and Funk  1988),  so expected  values  for  Yijk
are estimated from:

                      a   DJ   Vk   Bj,,   %s2
          E(Yijk)  =  e e   e   Xijk   e   ,                    (9)

where, s =  the  mean squared error from the general  linear  model.
The variance of individual predicted  Yijk is estimated from:

                       (2Yijk + a2)     a2
          Var(Yijk) =  [e         ] • [e  -  1]  .                  (10)

Although the above  expression  is  appropriate  when a  is  known
(Laurent 1963), s  is  assumed  to  be an unbiased  estimate of a for
the  1990 study since  only  a  small bias  was  introduced  into
estimates of the mean when s was used to  estimate a  in  past years
(Biggs and Funk 1988).

              Spawning Biomass per Billion Eggs  (B')

Catch sampling programs will be used  to estimate the relationship
between spawning biomass  and egg deposition. The tonnes of spawning
biomass required to produce one billion eggs (B') will be estimated
as:

                      W  • S
                 B' = —=— •  103  ,                           (11)
                       F(Wf)
where,
     W     = estimated average weight  in  grams  of  all herring
             (male and female) in the  spawning  population in an
             area,
     S     = estimated ratio of total  spawning  numbers (male and
     _       female) to female spawning numbers,
     F(Wf)  =  estimated fecundity at the average weight of females
             in the spawning population in an area,  in numbers of
             eggs, and
                                 10"6      grams  to tonnes
     103   =  conversion factor = 	 = 	.
                                 10"9      eggs to billions
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Estimates of  average  weight,  sex ratios, and fecundities  are  not
independent.  The variance of B'  is approximately:


     Var(B')  =  (103)2  {  [S/F(Wf) ]2 Var(W)

                      +  [W/F(Wf) ]2 Var(S)

                      +  [WS/F(Wf)2]2 Var(F(Wf))

                      +  2COV(W,S)  [S/F(Wf)] [W/F(Wf)]


                      -  2Cov[W,F(Wf)]  [S/F(Wf)] [WS/F(Wf)2]


                      -  2Cov[S,F(Wf)]  [W/F(Wf)J [WS/F(Wf)2] }.   (12)


The covariance terms containing S, Cov(W,S) and Cov[S,F(Wf) ], will
not be  included in the  estimate for  1990. These terms were  not
included in the estimate of Var(B') in  1988, 1989, and 1990  because
S was estimated from  either the  same pooled AWL  samples  or from a
single  AWL  sample. However,  Cov(W,S)  and Cov[S,F(Wf)] probably
contribute  a  small amount  to  Var(B') since the term  involving
Cov[W,F(Wf)] was very small in 1988, 1989, and 1990.

Correction  for Egg Loss:

The only  component needed for the  biomass estimate that has  not
been estimated within  the present study is egg loss (the proportion
of  eggs disappearing from spawning   areas  between the time  of
spawning  and  the time  of  surveys) .  Before the  extensive use  of
SCUBA diving  to survey  herring  egg deposition,  estimates of  egg
loss were  considered  to be  relatively high.  Montgomery  (1958)
estimated that  egg loss was 25  to  40% for southeast  Alaska,  and
Blankenbeckler and Larson  (1987) used similar estimates in their
early egg deposition surveys in southeast Alaska.  However,  Haegele
et  al.  (1981),  conducting  diving  surveys in  British  Columbia,
argued that egg loss was only about 10%. They based this assumption
on the  fact that most  spawn  was deposited  in  the subtidal zone
where egg loss, primarily  due  to  predation and  wave  loss,  was
probably  less than  had been  observed  in the  intertidal  zone.
Presently,  egg  loss  is assumed  to be 10%  in  British  Columbia,
southeast Alaska  and  PWS since  the timing of  diving surveys  in
relation to spawning  has been standardized among these  areas  (W.
Blankenbeckler,  ADF&G,   Ketchican,  pers.  comm.; Biggs  and Funk
1988).
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Herring Age, Weight, Length, Growth and Fecundity Estimation

Mean Weight and Sex Ratio:

Mean weight and sex  ratio will be  estimated from  AWLS  samples
collected  from  the  commercial catch  and  ADF&G  test  fishing
conducted before or after commercial openings.  AWLS samples will be
collected  from the spawning  population  in  each  of the  spawn
deposition summary areas (southeast, Valdez Arm, North Shore, Naked
Island,  and Montague  Island). The  approximate  timing of  peak
herring  spawning  in each  summary  area will  be  determined  from
aerial survey  sightings of  milt and  herring  schools. All herring
AWLS samples taken  during the  time of peak spawning in each area
will be pooled  to obtain estimates of mean  weight  and  sex ratio for
each summary area.  Mean weights and sex ratios for all of PWS will
be estimated as the average  of the estimates from  each of the areas
weighing by the spawn deposition biomass estimate in each area.

The estimated sex ratio, S,  is expressed as the ratio  of the number
of herring of   both sexes  in  the  AWL samples to the  number  of
females. The binomial  distribution  will be used  to  estimate the
proportion of females, p,  in samples, where S = 1/p. The variance
of S is then given by:

                   S2(S-1)
          Var(S) = 	,                               (13)
                      n

where,  n is the number of herring in the AWL sample.

Commercial  and test fishing  catches will be sampled  for  AWLS,
fecundity, and  roe  maturity  information. These data will be used to
estimate spawning  biomass and  spawn  deposition,  forecast herring
returns,  and  evaluate  effects of   the  oil   spill   on  survival.
Information on  fecundity, mean weight of females,  and  sex ratio are
also  important  components of the  spawn   deposition  biomass
estimator. AWLS sampling will  be intensified  in  1991 to increase
the precision  of biomass  estimates  and,  therefore,  enhance the
possibility of detecting oil spill impacts upon herring stocks.

Sampling will begin as soon as  concentrations  of herring appear in
nearshore areas that can be  sampled with purse seine gear.  Efforts
will be made to sample major concentrations of herring throughout
PWS at  periodic intervals throughout the spawning  period. The major
objective  of this  portion of the study will  be  to  determine the
age,  sex, and size composition  of all major herring concentrations
in the  general areas  including southeast area,  northeast  area,
North Shore area, Naked Island, and Montague Island. Results of the
aerial survey program will be used to direct test fishing efforts
within each area.
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Each week  during the sampling period, early  April through early
May, six to eight samples of herring will  be collected through test
fishing or  from  the  commercial  catch.  A  sample of 403 herring is
needed to  simultaneously  estimate the proportion  of  at  age of a
multinomial population  such that  95%  of the  time the estimated
proportions will be within ± 10% of the true proportions (Thompson
1987). Therefore, efforts will be made to  obtain samples consisting
of  approximately  450  herring  to  allow for  the  occurrence  of
unreadable  scales  (usually less than 5%  of the  sample).  Herring
samples will be flown from the fishing grounds each day to Cordova
for processing. Augmentation of the standard AWL sampling program
will  be needed  to  collect sufficient  samples  for  hydrocarbon
analyses, fecundity  estimates,  and oocyte loss measurements.  All
AWL data will  be collected using  personnel and  funding  from the
standard  (i.e.  non-oil  spill  related)  AWL  sampling  program
conducted by ADF&G within PWS.

The following data will be collected for each herring sampled:

     1. sex (determined by examination of gonads);
     2. standard length (in mm);
     3. weight (in grams);
     4. age (determined by examination of scales);
     5. capture information  (date  of capture,  fishing district,
        subdistrict, local name for the location,  fishing vessel
        name,  gear type);
     6. herring number on data form; and
     7. data form number.

Fecundity:

Additionally,  a subsample of herring will be collected to estimate
fecundity.   The  average fecundity at the  average female weight
(F(Wf)) from expression (11) is a component of  the spawn deposition
survey biomass estimator. The spawn deposition survey attempts to
estimate spawning  biomass  so that the  95% confidence interval is
within ± 25% of the actual  biomass estimate. If fecundity sampling
is to contribute no more than 1% to the confidence  interval width,
a sample of 85 females of  exactly the average  weight of females in
the spawning population is needed. Since average female weight is
unknown at the time of sampling,  more herring  must  be sampled over
a  range  of sizes.   Based  on  the precision of  1989  fecundity
sampling, a sample size of 130 herring would be needed to provide
the  desired  level  of  precision.    An   additional 100  samples
clustered around the mean size of females in 1991 will be taken to
compare with the past year's data. The mean weight  of a female in
the fecundity sample in 1990 was 131 grams. The predicted average
weight for  the population in  1990 is 155 grams, which translates to
an average predicted length of 230 to 240  mm.  Therefore, sampling
clustered about the 220 mm to 240 mm length classes is desirable.
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Effects of  the oil spill on  fecundity will also be  examined by
testing  for  differences  in   fecundity  among  five  areas:  (l)
southeast shore including Simpson and Sheep Bays, Port Gravina, and
Port Fidalgo; (2) northeast shore including Valdez Arm and Tatitlek
Narrows;  (3)  North  Shore;  (4)  Naked Island;  and  (5)  Montague
Island. While  extensive  mortality of adult herring  from  the oil
spill  has not  been documented,  it  is  possible that  sublethal
stresses could result in reduced fecundity.

Herring fecundity samples will be collected concurrently with AWL
samples. To accomplish this,  at  least five individual test purse
samples will  be  subsampled.   Females within  these purse  seine
samples will be randomly  selected within 10 mm length classes until
stratum goals are reached.  The roe sacs from each selected female
herring will be removed and placed in a Ziploc bag labeled with the
AWL number corresponding  to that female. Each individually packaged
roe sample will then be placed in a larger plastic bag labeled with
the sample date and location.  Standard laboratory procedures have
been developed to process fecundity samples.

Samples for  hydrocarbon analyses will also be obtained from herring
collected at each of the  four  locations (Naked Island, Galena Bay,
Cedar Bay, and Stockdale Harbor):

     1. three gut samples for hydrocarbons;
     2. three viscera samples for hydrocarbons;
     3. three muscle samples for hydrocarbons;  and
     4. three gonad samples for hydrocarbons.

General observations on the prevalence of nematodes,  liver and gall
bladder condition, and fullness of gut will also be made for each
herring collected  for hydrocarbon analyses.   Standard  protocol,
including sample  sizes  and  collection  strata,  for  collecting
herring eggs for hydrocarbon analyses will be followed.

In addition  to the  500 ovaries collected for fecundity analysis, 50
ovaries will  be collected and  preserved in a  buffered formalin
solution for oocyte loss measurements. An additional 25 preserved
ovaries will be obtained from Sitka Sound, southeast Alaska, for
use as a control.  Atretic eggs and histopathological damage in the
sac roe of the adult herring  will be recorded  during oocyte loss
observations.

A linear relationship was  found  between  fecundity and weight for
herring samples collected in 1988, 1989, and 1990 (Biggs and Funk
1988).  In 1991,  the fecundity-weight relationship  will again be
examined using data pooled  across all areas. Average fecundity for
each area  will be estimated from the  fecundity-weight relationship
using  the  average female  weight  from  each  area.  The  average
fecundity  for  each area  will_ then  be  applied  to  the  spawn
deposition biomass estimator (F(Wf) in expression (11) .  The variance


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of  estimated  average fecundities will be  approximated using the
variance  of  predicted means  from  the  fecundity-weight  linear
regression  (Draper and Smith 1981) :
             =s,[_  +  _  +         «— ,]•                 (14)
               2
where,

      s2  = residual mean square from the fecundity-weight
      _     linear regression,
      Wf  = average weight of female fish in the spawning
      	    population,
      WF  = average weight of females in the fecundity sample,
      W;   = weight of individual females in the fecundity sample,
      n   = total number of females in the fecundity sample,  and
      q   = total number of females in the AWL sample.

General Linear Model  (GLM)  extensions of linear ANOVA techniques
will be  used to test for  year and  area effects  in growth and
fecundity.

Egg Survival Study:

Oil  contamination  of herring  spawning  sites  and  exposure  of
spawning  herring to  oil may  cause  mortality to  herring eggs,
decrease  hatching success,  reduce  larval viability,  and impair
larval growth. The  major objective of  this  portion of the study
will  be  to  measure  immediate,  easily observable  mortality  of
herring eggs in  a subsample of the sites  used in 1989.  In 1991,
nine sites  will  be used  to conduct the  egg loss study, collect
hydrocarbon samples, collect live eggs  for the laboratory portion
of the study, and gather samples for sublethal impact testing.

Three study transects will be re-established in each of three areas
used during 1989  and  1990 (assuming those areas receive spawn in
1991): Naked Island, Fairmont Bay,  and Rocky Bay  on  north Montague
Island. The ratio of live to dead  eggs will be determined along
each transect from subsamples of 100 eggs. Dead eggs turn  an opaque
white color and are easily identified  with low  power magnification
under a binocular microscope. Mussel  tissue samples will also be
collected for hydrocarbon analysis.

Divers will establish the location of mean lower  low water (MLLW)
at the start of each dive.  Each dive team will attempt to sample
three transects each day.  Each  transect will be  sampled  every two
days until most herring eggs have hatched  (about  20 May). A total


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of twelve to  sixteen  dives  will  be made along each transect over
the course of egg development.

The location  of each transect will be marked. Divers  will work
along transects by following a compass  course set perpendicular to
shore. During the first dive,  five sample stations  at the +1, 0, -
5, -15,  and  -30  foot depths will be marked underwater with weighted
floats  anchored by a spike.  Station depths,   corrected  for tide
stage, will  be determined using diver's depth gauges. Three samples
of vegetation containing  at least 100 eggs will be  collected at
each depth along the transect whenever possible.

The following data will be recorded the first time each transect is
sampled:

     1.    transect number;
     2.    site description  (location, exposure, plant community);
     3.    number  of  depth  strata from which  herring  eggs were
          obtained; and,
     4.    original treatment  category  (high,  medium, low,  or no
          oil-impact).

The following data will be recorded every  time each transect is
sampled:

     1.    transect number and location;
     2.    date;
     3.    dive time;
     4.    treatment level;
     5.    air and water temperature;
     6.    maximum depth; and,
     7.    number of live, dead,  and other eggs per sample.

Herring  eggs  and  mussels  will  be  collected   at  each  site  for
hydrocarbon analysis on the first day. Three samples each of eggs
and mussels  (six per transect) will be collected from each sampling
location, including the three control sites  in  Sitka Sound,  at the
lowest tide stage at which mussels occur (usually about 5 ft below
MLLW).  Collection  methods  will  follow  established  protocol,
including chain of custody forms.

During one of  the sampling trips to each transect, herring eggs and
associated  vegetation  will  be  collected  for  the  laboratory
incubation project. Herring eggs  will  be  collected at nine sites
within PWS and three sites within Sitka Sound.  At each site, three
samples  of  vegetation  containing at  least  300  eggs  will  be
collected at three depths (MLLW,  -5 ft, and -15).

Herring eggs will  also  be  collected and preserved  in a phosphate
buffered  formalin  solution,  using  seawater,  for  biochemical
analysis. Results of these analyses may help determine the  extent
of oil exposure from determination of sublethal effects.

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Finally, herring egg samples will be collected from each of the 12
study sites for cytogenetic analysis. Ten egg patches consisting of
approximately 1000 eggs each (5 ml) will be preserved in a buffered
formalin  solution  from each  study site  (i.e.  a total  of  120
samples) . A subsample  of  eggs will be taken from each  sample  jar
and  analyzed  for mitotic  aberrations  in the  embryonic and yolk
cells.  Detailed methodology will be provided by the lab contracted
to perform the service.

Egg  survival  data  will  be summarized  by  level  of  hydrocarbon
impact, transect, depth, date of sample collection,  and proportion
of live eggs.  Several different analyses will be conducted to test
for differences in egg survival due to the level or amount of oil.
The  first  analysis  will  be  a   nested   mixed  factor   ANOVA
incorporating all possible  factors and  interaction  effects like:
Yijkl = u + AJ + Bj(Ai) + Ck + D, + ACfc + ADU + CDU + ACD^ + eijkl,      (15)

where ,

     Yijkl =  the  arc sin transformed  proportion  of live  eggs,
     u  =  grand  mean,
     A;  =  oil  impact  level (treatment;  fixed  effect) ,
     Bj  =  transect (random effect;  nested within treatment) ,
     Ck  =  depth  (fixed  effect) ,
     D,  =  time interval  (days) between spawning and sample
            collection (random effect) ,
     AClk + ADa + CDU  + ACDju = interaction terms, and
     eijkl =  error  terms,  which, after arcsine transformation are
            assumed to be  normally distributed  with mean  0 and
            variance a2 .

The second analysis will  be an analysis  of  covariance  (ANCOVA)
where  both  treatment  (A;)  and  time  (D])  will  be   treated  as
covariates. Treatment and  depth will be treated as fixed  effects,
while transect (nested within treatments)  and  time will be treated
as random  effects. This model will describe  the decrease in the
proportion of  live eggs over  time, using time as a covariate, and
will reduce the number of parameters that must be estimated for the
model.

Egg Loss Study:

Egg loss  is the  only component  of the  spawn deposition biomass
estimator that has not been measured. In the past, a 10%  egg  loss
factor was applied to  all transect data to adjust the total spawned
biomass  estimate.  In  1990  a  preliminary  egg  loss  study  was
conducted in conjunction with the egg survival  study to determine
whether the 10% egg loss factor is appropriate for use at PWS study
locations. The egg loss study will be continued in 1991.
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The same three transects used  in  each  of  three areas for the egg
survival study will be used  in the  egg loss study:  Naked Island,
Fairmont Bay, and Rocky Bay on north Montague Island.  Egg loss will
be estimated by observing changes in egg density over  time at these
locations.

To avoid sampler bias  in selecting samples a marked leadline, 20 m
or less  in  length,  will  be used to select  samples.  The leadline
will be placed parallel to shore and to the left of each transect
station. Egg density estimates will be taken within 0.1 m2 sample
quadrants using the same procedures described for spawn deposition
diver transects.  For each transect, five egg density estimates will
be made  at  each  of five  depths  (+1,  0,  -5,-15,-30  ft  depths).
Divers making egg density  estimates  for the egg  loss study will be
calibrated in a similar manner used for divers assisting in spawn
deposition  surveys. One  egg  count  calibration  sample will  be
collected at each  transect and  at  each depth  level.  For  the
calibration sample,  all herring eggs and vegetation will be removed
from  a  0.1  m2   sample   quadrant.  Counts  of  eggs  within  the
calibration sample will be made in the  laboratory at a later time.
Egg density estimates  and  egg counts will be conducted every other
day from the time  of spawning in  each area  until  the  time  of
hatching  (a  period of approximately   20-25  days).  It  should  be
possible to obtain egg density estimates and egg counts for about
eight  days  during  the  study.  This would result  in a  total  of
approximately 1,800 egg density estimates (three areas; 3 transects
per area; five depths  per  transect;  five egg density estimates per
depth; eight days) and 540 egg counts (three areas; three transects
per area; five depths  per  transect;  one egg count per depth; eight
days)  for the season.

Egg loss data will be  summarized by area, transect, depth, date of
sample collection, and estimated egg density.  Egg density estimates
will be adjusted for observer (diver) biases, following procedures
set forth for diver calibration in the spawn  deposition survey,
prior to analyses.  The change in egg  density  over time for each
transect and depth will be examined.

Egg Incubation Experiment:

A much smaller laboratory egg incubation experiment will be carried
out by a private consultant  contracted by ADF&G.  This experiment
will estimate the survival of herring  eggs and  larvae collected in
PWS in  1991. The preliminary  results  of the   1989  and  1990 egg
incubation experiment can be found in McGurk et al.  (1990).

The objective of  the 1991 experiment is to  replicate the experiment
done in  1989  and 1990 but on  a much  smaller scale.  The eggs and
larvae will  be reared under the same  conditions  as  they were in
1989  and  1990.   The   eggs  and  herring  collected  during  this
experiment will  be sent  to  another   independent  contractor for
sublethal testing.  The results from  the sublethal  testing will

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allow us to compare sublethal effects in 1989, 1990,  and 1991 under
the same laboratory conditions.

Oil Exposure Study  (Dose-Response):

The major  addition to the 1991 herring study is an oil exposure
study that will measure the effects of oil exposure on herring eggs
and larvae.
                           BIBLIOGRAPHY

Biggs, E.D.,  and F.  Funk. 1988.  Pacific  herring spawning ground
     surveys  for  Prince  William  Sound,  1988,  with  historic
     overview.   Regional   Information  Report   2C88-07,   Alaska
     Department of Fish and Game, Anchorage, 73 p.

Blankenbeckler, W.D. and R. Larson. 1982. Pacific herring  (Clupea
     harengus pallasi)  spawning ground research  in southeastern
     Alaska,  1978, 1979,  and  1980.  Alaska Department of Fish and
     Game Technical Report No. 69. 51 p.

Blankenbeckler, W.D. and R. Larson. 1987. Pacific herring  (Clupea
     harengus pallasi) harvest statistics,  hydroacoustical surveys,
     age, weight, and length analysis,  and spawning ground  surveys
     for southeastern Alaska,  1980-1983. Alaska Department  of Fish
     and Game Technical Data Report No. 202. 121 p.

Cochran, W.G. 1963. Sampling techniques. John Wiley and Sons, New
     York.

Draper, N.R. and H.  Smith. 1981.  Applied regression analysis. John
     Wiley and Sons, New York.

Goodman,  L.A.  1960.  On the  exact variance  of  products.   J.  of
     the Amer. Stat. Assoc. 55:708-713.

Haegele,   C.W.,   R.D.  Humphreys,   and  A.S.   Hourston.   1981.
     Distribution of eggs by depth and vegetation type in  Pacific
     herring  (Clupea  harengus  pallasi)  spawnings  in  southern
     British Columbia. Can. J. of Fish. Aquat. Sci. 38:381-386.

Jackson,  M.   and  R.C.  Randall.   1983.  Herring  spawn  deposition
     surveys  in Prince  William Sound,  1983.  Alaska Department of
     Fish and Game, Prince William Sound Data Report No. 83-6. 15p.

Jackson,  M.   and  R.C.  Randall.   1984.  Herring  spawn  deposition
     surveys, Prince  William  Sound, 1984.   Alaska Department of
     Fish and Game, Prince William Sound Data Report 84-16. 15 p.
                               153

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Laurent, A.G.  1963.  Lognormal  distribution and  the  translation
     method: description and estimation problems. J. of the Amer.
     Stat.  Assoc. 58:231-235.

McGurk, M.,  D.  Warbuton,  T.  Parker, and M. Litke.  1990. Early life
     history of Pacific herring:  1989 Prince William Sound herring
     egg incubation experiment.   Final Report  prepared for NOAA,
     National   Ocean    Service/OMA/OAD.    Triton   Enviromental
     Consultants Ltd.,  Richmond, B.C.,  Canada.  73 p.

Montgomery,  D.T.  1958.  Herring spawning surveys  in southeastern
     Alaska. United States  Fish and Wildlife  Service,  Bureau of
     Commercial Fisheries,  Marine Fisheries Investigations Field
     Operations Report. 22 p.

SAS Institute  Inc.  1987. SAS/STAT Guide for personal computers,
     version 6 edition. SAS Institute,  Gary, North Carolina.

Schweigert,  J.F., C.W. Haegele,  and M.  Stocker.  1985. Optimizing
     sampling design for herring spawn surveys on  the Strait of
     Georgia,  B.C. Can. J. of Fish, and Aquat.  Sci.  42:1806-1814.

Thompson,  S.K.  1987.  Sample   size  for  estimating  multinomial
     proportions. The American Statistician 41:42-46.
                              BUDGET

Salaries                    $ 238.5
Travel                          5.5
Contracts                     299.3
Supplies                        9.7
Equipment                       5.0

Total                       $ 558.0
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FISH/SHELLFISH STUDY NUMBER 13

Study Title:  Effects of Hydrocarbons on Bivalves

Lead Agency:  ADF&G


                           INTRODUCTION

Bivalve mollusks  are an important  component of the  food chain,
existing as prey for bear and sea otters, and support subsistence
and sport fisheries in PWS.  Because they are relatively sedentary
and  occupy   nearshore  areas,   bivalves  may   be  particularly
susceptible  to  contamination   by   oil.     Bivalves  metabolize
hydrocarbons  at  a   slow   rate   and  are   therefore  likely  to
bioaccumulate hydrocarbons.   It is hypothesized  that increased
hydrocarbons in nearshore  sediments could affect  bivalves  for a
long period of time by increasing mortality, decreasing growth, or
causing sublethal injuries.  The effects of oil on the growth and
survival of littleneck  clams  (Protothaca staminea) in particular
and other bivalves in general have been well documented (Anderson
et al.  1982; Anderson et al. 1983; Augenfeld et al.  1980; Dow 1975;
Dow 1978; Keck et al. 1978).

This study  is  a  continuation of work which  was conducted during
1989 and 1990.  During  1991 field work  will be  conducted only in
PWS.  Clam aging,  data  entry and analysis from 1989 and 1990 will
continue.


                            OBJECTIVES

A.   Test if the level of hydrocarbons in bivalves and in sediments
     is not related to  the level of oil contamination of a beach.

B.   Document the presence and type  of damage to tissues and vital
     organs of bivalves sampled from beaches such that differences
     of ± 5%  can  be determined  between  impact  levels  95% of the
     time.

C.   Test if the  growth rate  of littleneck clams  is  the same at
     beaches of no oil  impact, intermediate or high levels of oil
     impact.

D.   Identify potential alternative methods  and  strategies  for
     restoration of lost use,  populations,  or habitat where injury
     is identified.
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                             METHODS

The  1991  field portion of  this study will  be conducted  by the
ADF&G.   Field work  will  be limited  to  a reciprocal  transplant
involving  littleneck  clams.    A  similar  transplant  study  was
conducted in  1990.   During April 1991, clams will  be  tagged and
transplanted between the same oiled and unoiled sites utilized in
1990.   These  sites are located  in  the vicinity of  bear  and sea
otter habitat.

Six study sites for littleneck clams in PWS representing two levels
of oil  contamination (no  contamination and  intermediate  or high
contamination) will be sampled.

For each sample site,  the  following site description information
will  be  recorded:     site  orientation  (N-NW  etc.),   latitude,
longitude, low tide height,  temperature and salinity  of the water,
weather and wave action.    Temperature and salinity of the water
will be measured at a distance of approximately 5 m offshore from
the sampled beach at the daily low slack tide.

To quantify oil  impacts  on  clam  growth  and  to discount  site
effects,  littleneck clams  will be transplanted  from oiled  to
unoiled areas and from unoiled to oiled areas. Three  oiled beaches
and three unoiled beaches were chosen for this purpose.  Criteria
for selecting paired  oiled/unoiled beaches, to the extent possible,
will include similarity in profile,  drainage and length-frequency
distribution of bivalves.

Two tidal heights will be utilized,  each of  which has an adequate
number of specimens at  paired beaches.  Clams will be transplanted
to the same  tidal height from which they originated.   At each tidal
height, three stations will  be established  creating  triplicate
sampling stations at each height.   Each  location will  consist of
three adjacent clearly marked  0.25  m2 plots.   One  plot  will be
marked, but will  not  be  disturbed until clams are sampled for
growth.  Another plot will  be  dug to  a depth of 0.3 m  and all of
the removed clams and sediment will be replaced in the plot.  Clams
from this plot will have a  small notch  filed  into the ventral edge
of the valves to mark the time of disturbance.  All  clams will be
removed from the third plot which will be dug to a depth of 0.3 m
and the transplanted clams will be placed in this plot along with
the original sediment.

Clams to be  transplanted will be obtained by digging  a trench along
the prescribed tidal height of  the donor beach  until  150 clams
between 15  mm and  35 mm in length  have  been collected.   Fifteen
millimeters  is considered  to  be   the smallest  size  which can
effectively be tagged.   Clams less than 35 mm  are selected to
narrow the range  of ages for which differences in growth are being
determined  and because  the maximum growth rate appears  to occur
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within this  size range.  A  sample of 50 specimens  from each of
three plots will provide 150 samples from each tidal height at each
beach  and 450  clams  for each  tidal height  and  level  of  beach
impact.  Sample size for growth  is  based on the difference between
mean shell height  for  age i  and age i+1  clams,  variance in shell
height for  age i+1 clams, probability of making  a  type I  error
equal to  0.01  and  probability of making  a type  II error equal to
0.05 (Netter and Wasserman 1985).  The sample size was determined
after comparing data for mean shell height and  variance in shell
height taken from Paul and Feder (1973) and Nickerson  (1977) .  The
sample size  for detecting  between  impact level  differences  in
growth at age  of clams  in the  size range of 15 mm  to  35  mm was
estimated at 133 clams  from the Paul and Feder data and at 85 clams
from the Nickerson  data for each impact level.  The  higher estimate
was rounded  up to 150  clams  by including the  next  smaller size
group (age 5-6).  The purpose of 3 sites for each  impact level is
to provide replicates at each impact level.

Transplanted clams will be identified by marking each clam with a
numbered Floy tag secured with a quick-drying adhesive. All marked
clams will have a  small notch filed into the ventral edge of the
valves to mark the  time of transplantation.  Individual clams will
be  measured  at  the beginning  and  end  of the  experiment.   In
September of 1991,  near the end of the growing season, clams will
be removed from each of the plots described above and  analyzed for
growth.  Wet and dry weights of clams will also be recorded so that
clam condition can be  compared in  terms  of  a weight  to  length
ratio.  Hydrocarbon and histopathology samples will be taken during
the experiment.

A total of six sediment samples will be  collected from each site
for hydrocarbon  analysis.   The  triplicate  sediment  samples from
each tide height will be composite  samples which will  be  collected
by scooping one tablespoon of sediment to a depth of 2 to  3 cm from
each of  the  nine sample quadrates  at  a  tide height.   The small
subsamples of  sediment  taken  from  each  sampling quadrate will
provide  a representative  mixture  of sediment composition  and
contamination along the tide height.

Two hydrocarbon tissue samples will be obtained from each sampling
station.   Each hydrocarbon sample  will  be composed  of  10  to 20
clams.  Specimens with  a shell length of 2 - 5  cm will  be  collected
from the donor beach concurrent with the collection of clams for
tagging   to   form   a    hydrocarbon  sample  at   the   time   of
transplantation.  During transplantation  10 to 20 additional clams
will be collected  from the donor beach  for placement with tagged
clams in  quadrate  "A"  at each sample station.   These clams will
comprise the hydrocarbon sample during fall recovery.

Combined tissue samples from each sampling station will provide a
representative  mixture  of  bivalve   tissue   composition  and
contamination across the site.   The desired size of each  composite

                               157

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tissue sample is 15 grams.  The number of bivalves to provide this
sample from each transect was estimated based on the average size
of individuals of each species.

Collection  of  specimens  for  necropsy  will  begin  after  all
hydrocarbon samples have been taken.  Total sample size is 20 live
or  moribund specimens  taken at random  from  each beach  site.
Noticeable  numbers of  moribund  animals will  be documented  and
sampled separately.

To  address  Objective A (hydrocarbons  in  sediments and  bivalve
tissues) ,  an ANOVA will  be used  to  test  for  differences  in
hydrocarbon  content in  sediment between  sites.  Differences  in
sediment hydrocarbon content  will verify that control sites (areas
of no oil impact) are in fact "controls".   These differences will
also permit post-stratification of sample sites according to level
of  impact.   An  analysis of  variance  will  be  performed on  the
hydrocarbon content of  clam  samples among sites.  The  results of
this test will be related to the level of sediment impact.

Objective  B  will  be  met   through  ANOVA  contingent  upon  the
processing of necropsy samples.  These samples will be processed if
hydrocarbon analysis is positive.

To provide baseline (pre-impact)  information on variance in growth
at age among sites, an  analysis  of variance  on  growth  parameters
from  clams  taken  during  1989 between  areas  will  be  conducted.
Growth parameters  will  be determined for various growth  curves,
such as Gompertz, von Bertalanffy, or polynomial  equations.  Growth
parameters will be presented for the most appropriate growth models
only.  A similar ANOVA will be conducted on growth parameters from
clams taken during 1990  between areas.   Those beach sites which are
resampled in 1990 will be subjected to an analysis of variance on
growth parameters obtained from fitting  algorithms for clam growth
after  impact (1990 and  beyond)  and will  be compared  to growth
parameters  for  clam growth prior to impact  (approximately  1979-
1989) to resolve impact of oil contamination on growth (Objective
C).  Graphics will be used to display differences in growth among
areas over time, including growth curves (size at age)  and growth
increment at age by year for each beach.

To address  Objective  D, all  data  will be analyzed to determine
degree of damage to stocks.  Appropriate suggestions will be made
for  restoration  or  mitigation  measures.     This  may  include
restrictions on  human usage  to  reduce exposure  to carcinogenic
levels of hydrocarbons or to protect threatened clam populations.
Other  actions  may  include the need  for continued  monitoring of
stocks.
                               158

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                           BIBLIOGRAPHY

Anderson, J.W., R.G. Riley, S.L. Kiesser, B.L. Thomas, and G.W.
     Fellingham.  1983.    Natural  weathering  of  oil in  marine
     sediments: tissue contamination and growth of the littleneck
     clam, Prototheca staminea.  Can. J. of Fish, and Aquat. Sci. 40
     (Suppl. 2):70-77.

Anderson, J.W., J.R.  Vanderhorst,  S.L. Kiesser, M.L. Fleishmann,
     and G.W.  Fellingham. 1982. Recommended methods for testing the
     fate and effects of dispersed oil  in marine sediments.  In Tom
     E. Allen  (ed.),  Oil  spill chemical  dispersants:   research,
     experience,  and  recommendations.    ASTM  Special  Technical
     Publication 840.  Philadelphia, Pa. p. 224-238.

Augenfeld, J.M., J.W. Anderson, D.L. Woodruff, and J.L. Webster.
     1980. Effects of Prudhoe Bay crude oil-contaminated sediments
     on  Protothaca  staminea  (Mollusca:Pelecypoda):  hydrocarbon
     content,  condition index,  free  amino  acid level.  Marine
     Environmental Research. 4(1980-81):135-143.

Dow, R.L. 1975. Reduced growth and survival of clams transplanted
     to an oil spill site. Marine Pollution Bulletin. 6(8):124-125.

Dow, R.L.  1978.  Size-selective mortalities of clams in an oil spill
     site.  Mar. Poll. Bull. 9(2):45-48.

Keck,  R.T.,  R.C.  Heess,   J.  Wehmiller,  and  D.  Maurer.  1978.
     Sublethal effects of  the water-soluble fraction of nigerian
     Crude Oil  on the Juvenile Hard  Clams,  Mercenaria  (Linne).
     Environmental Pollution. 15:109-119.

Neter,  J., W. Wasserman, and M. Kutner. 1985.  Applied Linear
     Statistical Models. Richard D. Irwin, Homewood Illinois.

Nickerson, R.B. 1977. A Study of the littleneck clam Prototheca
     staminea  (Conrad)  and the butter clam,  Saxidomus giganteus
     (Deshayes) in a habitat permitting coexistence, Prince William
     Sound, Alaska.  Proceedings of the  National Shellfisheries
     Association. 67:85-102.

Paul, A.J. and H.M. Feder.  1973.   Growth, recruitment, and
     distribution of  the littleneck clam,  Protothaca staminea in
     Galena  Bay,   Prince   William  Sound,  Alaska.    Fish.  Bull.
     71(3):665-677.
                               159

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                              BUDGET

Salaries                    $  88.0
Travel                          5.0
Contracts                      50.0
Supplies                        2.0
Equipment                       2.0

Total                       $ 147.0
                               160

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FISH/SHELLFISH STUDY NUMBER 27

Study Title:  Sockeye Salmon Overescapement

Lead Agency:  ADF&G


                           INTRODUCTION

Commercial  fishing  for  sockeye salmon in  1989,  was curtailed in
upper Cook Inlet (CI) ,  the outer Chignik districts,  and the Kodiak
areas due to presence of  oil  in the fishing areas from the EVOS.
As a result, the number of sockeye salmon entering four important
sockeye producing systems (Kenai/Skilak,  Chignik/Black,  Red,  and
Frazer Lakes)  and two  less  important lake  systems (Akalura  and
Afognak or Litnik lakes) greatly exceeded levels that are thought
to be most  productive.    Sockeye salmon  spawn in lake-associated
river systems.  Adult salmon serve an extremely important role in
the  ecosystem,  providing  food  for  marine  mammals,  terrestrial
mammals,  and birds.  Additionally, carcass decomposition serves to
charge freshwater lake systems with  important nutrients.  Juvenile
salmon which rear in  lakes for one or two  years  serve  as a food
source for  a variety of fish,  birds and  mammals.   Sockeye salmon
are also an important subsistence,  sport, and commercial species.
The ex-vessel value of the commercial catch of sockeye from these
lake systems has averaged about $42  million per year since 1979,
with the 1988 catch  worth $115 million.  Sockeye salmon returns to
the Kenai River  system support some  of  the largest recreational
fisheries in the State.

Overly large  spawning  escapements  may result in  poor returns by
producing more rearing  juvenile sockeye  than can be supported by
the nursery lake's productivity (Kyle et  al. 1988).  In general,
when rearing  fish  abundance greatly  exceeds  the  lake's carrying
capacity, prey resources are altered by changes in species and size
composition (Mills and Schiavone 1982; Koenings and Burkett 1987;
Kyle et al. 1988) with  concomitant  effects on all trophic levels
(Carpenter et al. 1985).  Because of such changes,  juvenile sockeye
growth is reduced, mortality increases, larger percentages holdover
for  another year  of rearing,  and  the  poor  quality  of  smolts
increases marine mortality.   Where escapements are two to three
times normal levels, the resulting high juvenile  densities crop the
prey resources to the extent that more than one year  is required to
return to normal productivity.  Rearing juveniles from subsequent
brood-years suffer from both  the poor quality of forage and from
the increased competition for  food by  holdover juveniles (Townsend
1989).   This  is the   brood-year  interaction underlying  cyclic
variation in the year class strength of anadromous fish.

This  project  will  examine the effects  of large  1989  spawning
escapements on the  resulting  progeny for a select  subset of the
above mentioned sockeye  nursery lakes.  Three impacted lake systems

                                161

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where the 1989 escapements were more than twice the desired levels
(Kenai/Skilak in Upper CI; Red and Akalura lakes on Kodiak Island)
were selected.  Upper Station Lake which is near the two impacted
lakes on  Kodiak did not  receive a  large escapement and  will be
examined as a control.

This study  is necessary  to obtain a  more timely  assessment of
impact,  as adult sockeye produced from the 1989 escapement will not
return until the 1994/1995 season.  Further, total return data are
not available  for individual Kodiak  sockeye  systems due  to the
complex mixed-stock  nature of the  commercial  fisheries  and the
inability to estimate stock-specific catches.


                           OBJECTIVES

A.   Estimate the number,  age, and size of sockeye salmon juveniles
     rearing in selected freshwater systems.

B.   Estimate the number,  age, and size  of  sockeye salmon smolts
     migrating from selected freshwater systems.

C.   Determine effects of large escapements resulting from fishery
     closures  caused by  the EVOS  on  the rearing capacity  of
     selected nursery lakes through:

     1.    analysis of age and growth of juveniles and smolts
     2.    examination of  nursery area nutrient budgets and plankton
          populations.


                             METHODS

Numbers of  adult sockeye  salmon  that entered selected spawning
systems outside PWS prior to and during 1989  have been estimated at
weir stations or by sonar.  This information was collected during
projects routinely conducted by the ADF&G as part of their resource
management  program.    Optimal escapement levels,  which  on the
average should produce maximum sustained  yield, have been based on
either past relationships between spawners and returning progeny or
the extent of available spawning and rearing habitat.  The baseline
program will continue at  each site,  including  but not limited to
estimates of adult sockeye escapement  and collection of scales for
age analysis.

For each of  the 4 lake systems identified,  the response (abundance,
growth,  and freshwater age) of  rearing  juveniles  from the 1989
escapement will be studied through its  likely period of freshwater
residence, early summer 1990 to spring 1992.

The total number of juvenile sockeye in each lake will be estimated
through hydroacoustic  surveys  conducted during the summer  (late

                               162

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June)  and fall  (September-October)  of 1990, 1991,  and possibly
1992.   Age and  size  information as well  as diet  items  will be
obtained from samples of juvenile sockeye collected from concurrent
mid-water  trawl  netting  surveys.    Survey transect  designs for
hydroacoustic sampling and  tow-netting have been established for
Kenai  and  Skilak  lakes  (Tarbox  and  King  1989) ,   and will  be
developed for each additional lake in the study.   The basic survey
design  will be  a  stratified  random sample where  each  lake is
subdivided  into  areas  and survey transects  randomly selected in
each  area.    Such programs,  funded through other  studies,  are
already in  place for  Tustumena and Afognak  lakes.   Depending on
densities of rearing juvenile sockeye,  estimates of  fish densities
will be made  for each transect either by  echo  integration or by
echo counting.  Total fish population estimates will be computed,
by  summing  transect  populations,  along  with  95%  confidence
intervals (Kyle 1989).

Freshwater growth and age of sockeye  salmon rearing  juveniles from
all  study  systems  will  be  determined  from  scale  and  otolith
measurements made either by direct visual analysis of scales or on
an Optical  Pattern  Recognition system.  In  cases where data are
available (e.g., Kenai and  Skilak Lakes),  growth of progeny from
the 1989 spawning escapements will be compared with growth (size)
of progeny  produced from  spawnings  within  these systems during
prior years.

The  total number of  smolt  migrating  from  each  system  will be
estimated  with  a  mark-recapture  study during  1990,  1991,  and
possibly  1992  using  inclined plane traps  after Kyle (1983), and
Tarbox and King  (1989) .  Smolt will be captured in traps, sampled
for  age and  size  information,  marked with Bismark Brown  Y (a
biological dye) ,  and transported upstream of the traps and released
for subsequent recapture  (Rawson  1984).   Periodic retesting will
determine the capture efficiency of the traps under  changing river
conditions during the spring.  Total population estimates (with 95%
confidence  intervals)  will be  made  using catch  efficiencies.
Weekly  number weighted smolt size and  age information  will be
calculated using a computer spreadsheet developed by Rawson  (per.
comm.  1985) .    Size and  ages of  sockeye  smolts from  the  1989
spawning escapements will be compared with smolt  information from
spawnings within these systems during prior years.   Finally, smolt
programs consistent to those for the study lakes are  planned, under
separate funding, for Tustumena and Afognak Lakes.

Limnological studies will monitor the response of  the  lakes to the
high  juvenile rearing  densities  and  to  estimate   the carrying
capacity parameters of euphotic volume, nutrient budgets  (carcass
enrichment), and zooplankton biomass,  body-sizes, and population
shifts.  Approximately six limnology surveys will be conducted at
two stations, during 1990,  1991,  and possibly  1992, to determine
zooplankton species abundance and body-sizes, nutrient chemistry,
and phytoplankton abundance for Kenai/Skilak,  Red, Akalura, and

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Upper  Station  lakes.    Carrying-capacity  parameters  exist  for
Afognak and Tustumena lakes based on ongoing studies by ADF&G FRED
and Commercial Fisheries Divisions.

In cases where seasonal data are available  (e.g., Akalura, Kenai,
and  Skilak lakes),  limnological  parameters  taken during residence
of  the juveniles  from  the 1989  spawning  escapements   will  be
compared to parameters within these systems  during prior years.

The holistic  approach proposed here  involves  several  evaluation
procedures to assess the effects  of sockeye  salmon overescapement.

First, freshwater  production from the  1989 escapements  will  be
assessed in Kenai/Skilak,  Red, Akalura,  and Upper  Station lakes.
This will be  accomplished  through analysis  of growth,  freshwater
survival (in particular overwinter survival), and freshwater age of
sockeye smolt  populations.   Any anomalies  will  be  determined by
analysis  of  freshwater  growth  recorded  on  archived  scales,
historical  freshwater age  composition,  and  modelled  freshwater
survivals; and from results of previous studies as well as  the 1991
smolt  characteristics  from  each of  the study  systems.    Also,
planktonic  food  sources will  be assessed  through  estimation  of
abundance of zooplankton prey biomass and numbers of species.

Second, future sockeye salmon production  from the 1989 parent year
and   subsequent   parent  years   will  be  estimated  based  on
spawner/recruit   relationships    incorporating    a   brood-year
interaction  term.     Losses of   adult   sockeye  production  from
subsequent parent years may result from negative effects of progeny
of  the 1989  escapement  on the lake's  carrying  capacity.  The
spawner/recruit  relationships  will be estimated from  historical
stock  specific  return  data (where  available),  and  generalized
spawner/recruit  data  scaled to  the carrying  capacity  parameters
(i.e., euphotic  volume and  zooplankton  biomass) of the  nursery
lakes where stock specific return data  are  not available  (Geiger
and Koenings 1991).

Third, experimental and empirical sockeye life history/production
models (Koenings and  Burkett 1987; Koenings et al.  1989)  will be
used  to  compare salmon production by  life-stage  at  escapement
levels consistent with management goals to the 1989 escapements.
                           BIBLIOGRAPHY

Carpenter,  S.  R.,  J.  F.  Kitchell,  and  J. R.  Hodgson.   1985.
     Cascading   trophic   interactions  and   lake  productivity.
     BioScience 35:634-639.

Geiger, H.  J.,  and J.  P. Koenings.  1991.   Escapement goals for
     sockeye salmon with informative prior probabilities based on
     habitat considerations.  J. of Fish. Res. (in press).

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Koenings,  J.  P.,   and   R.   D.   Burkett.     1987.    Population
     characteristics of sockeye salmon (Oncorhynchus nerka) smolts
     relative to temperature regimes, euphotic volume,  fry density,
     and forage  base within  Alaskan Lakes, p.  216-234.  In H. D.
     Smith,  L.  Margolis,  and C. C.  Wood  (eds.)  Sockeye salmon
     (Oncorhynchus nerka) population biology and future management.
     Can. Spec. Publ. Fish. Aquat. Sci. 96.

Koenings, J. P., J. E. Edmundson, G. B. Kyle,  and J. M. Edmundson.
     1987.   Limnology field  and laboratory  manual:  methods for
     assessing aquatic production.  Alaska Department of Fish and
     Game, FRED Division Report  Series No. 71:212 p.

Koenings, J. P., R.  D. Burkett,  M. Haddix,  G. B.  Kyle,  and D. L.
     Barto.  1989.   Experimental manipulation  of lakes for sockeye
     salmon  (Oncorhynchus  nerka) rehabilitation and enhancement.
     Alaska  Department of Fish and  Game,  FRED  Division Report
     Series No. 96:18p.

Kyle, G. B.  1983.   Crescent Lake sockeye salmon smolt enumeration
     and sampling,  1982.   Alaska Department of Fish and Game, FRED
     Division Report Series No.  17:24 p.

Kyle,  G.  B.  1989.    Summary  of acoustically-derived population
     estimates  and  distributions  of  juvenile  sockeye  salmon
     (Oncorhynchus  nerka)  in 17 nursery  lakes  of  southcentral
     Alaska.  Alaska Department of Fish and  Game, FRED Division
     Report Series No. (In review).

Kyle, G. B., J.  P.  Koenings,  and B. M.  Barrett.  1988.   Density-
     dependent,  trophic  level responses to an introduced run of
     sockeye salmon (Oncorhynchus nerka)  at  Frazer  Lake, Kodiak
     Island, Alaska.  Can. J.  of Fish, and Aquat. Sci. 45:856-867.

Mills,   E.  L.,  and A.  Schiavone, Jr.   1982.   Evaluation  of fish
     communities   through  trophic   assessment  of  zooplankton
     populations and measures  of lake productivity.  N. Amer. J. of
     Fish. Mgt. 2:14-27.

Rawson,  Kit.    1984.   An estimate of  the size  of  a  migrating
     population  of  juvenile  salmon  using  an   index  of  trap
     efficiency obtained by dye marking. Alaska Department of Fish
     and Game, FRED Division Report Series No.  28:23 p.

Tarbox, K.E., and B.E. King.   1989.  An estimate of juvenile fish
     densities in Skilak  and Kenai Lakes, Alaska through the use of
     dual beam hydroacoustic techniques in 1989. Alaska Department
     of  Fish   and  Game,  Commercial  Fish   Division  Regional
     Information Report No. 2S90-1.

Townsend, C.R.  1989.  Population cycles in freshwater fish. J. of
     Fish Bio. 35  (Supplement A):125-131.

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                              BUDGET

Personnel Services           $189.7
Travel                         11.2
Contractual                   101.4
Supplies                       29.6
Equipment                       2.4

Total                        $334.3
                               166

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FISH/SHELLFISH STUDY NUMBER 28

Study Title:   Salmon   Oil    Spill    Injury   Model   and   Run
               Reconstruction

Lead Agency:   ADF&G
                           INTRODUCTION

This study integrates results obtained  from Fish/Shellfish Studies
1-10 to  determine damages  to wild Pacific  salmon (Oncorhynchus
spp.) resources  exposed  to crude oil  from the  EVOS which spread
through  portions of  PWS,  CI, Kodiak,   and  Chignik.   Damages to
Pacific  salmon populations  in these  areas  would  have  profound
impacts  on both  aquatic  and terrestrial ecosystems since Pacific
salmon are an important food source for many fish, bird, and mammal
species and cycle significant amounts of nutrients from marine to
estuarine, freshwater, and  terrestrial environments.    Also, the
economies and culture of many communities in this portion of Alaska
rely heavily on harvesting Pacific salmon in commercial, sport, and
subsistence fisheries.

Two  different procedures  may be  used in this study to assess
damages to wild Pacific salmon populations resulting from crude oil
contamination. The first, based on reconstructing salmon runs will
use  total  adult  returns  (harvests and spawning  escapements)  to
determine  stock  specific  returns  and production  to oiled and
unoiled  areas.   The  second,  based on  life history modeling, will
use  spawning  escapements and subsequent estimates  of survival at
various  life  history stages  to  project future adult  returns to
oiled and unoiled areas.   Both approaches will  use data from F/S
studies  1-10,   as   well   as  information  from  the  scientific
literature, to set parameter values in computational models.
                            OBJECTIVES

                        Run Reconstruction

A.   Develop  a  computational  framework  for  estimating  stock
     specific abundance over time in the eight commercial fishing
     districts in PWS.

B.   Analyze the historical data to  develop estimates of the model
     parameters,   including    estimates    of   hatchery   stock
     contributions.

C.   Reconstruct the 1990 and 1991 PWS  pink salmon run and develop
     estimates of salmon  production (number  of adult returns per
     spawner) for oiled and unoiled areas.
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                      Life History Modeling

A.   Develop a  computational framework  to account for  specific
     effects of oiling on species, stock, and life history stages
     of wild Pacific salmon (Oncorhynchus spp.) populations in PWS,
     CI, Kodiak, and the Chignik areas.

B.   Estimate "status quo" (i.e. in absence of oil contamination)
     values  for all  parameters  implicit  in  the  computational
     framework.

C.   Estimate the "oil impact" values for all parameters implicit
     in the computational framework.

D.   Develop estimates of salmon injury by comparing simulations of
     future Pacific salmon production using "status quo" and "oil
     impact" model parameter values.


                             METHODS

                       Run Reconstruction

This   portion   of  the   study   will   develop   techniques   for
reconstructing  stock  specific pink  salmon abundance  by  fishing
district in PWS.  The  study will consist  of three activities, data
synthesis, model development, and parameter estimation.

Data Synthesis.   Historical catch, effort, escapement, and tagging
data will be synthesized and an RBASE data base management system
developed to provide  easy access  to this data.   Details  of this
data are as follows:

A.   Catch data will be summarized by species, district, daily or
     biweekly time periods,  separated into hatchery and wild stock
     components for the years 1960 to 1991. Hatchery contributions
     from  1987  - 1991 will  be based on CWT tagging.   Hatchery
     contributions prior  to  1986  will be based  on assumption of
     equal  exploitation  rate  within  and  relative  escapement
     magnitudes by district.

B.   Effort  data will  be  summarized by district  on daily  or
     biweekly time periods for the years 1960 to 1991.

C.   Timing  curves  describing the  entry of escapement  into the
     stream will be estimated by stream within district for years
     1960  -  1991.     The  parameters of  the timing  curve  will be
     estimated by fitting the stream  life model of live fish in the
     stream to the escapement counts expanded to areas not counted
     by aerial survey. The  stream specific  expansion  factors will
     be based on comparison of on-ground counts to aerial counts.

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D.   Extensive  and   comprehensive   tagging  studies  have  been
     conducted  in  PWS since 1957.   A database management system
     will be developed to summarize those data.  A database will be
     used to estimate parameters of stock specific migration models
     (see model development section below).

                        Model Development

The model below is developed in  full  generality.  In estimating
model parameters it  may  be  necessary to simplify the model.  The
following definitions and relationships apply:

indices:

     a    =    fishing district,  (eight districts)
     s    =    stock,  (eight wild stocks, four hatcheries)
     t    =    time

     N,M  =    abundance of stock s in district a
     XM   =    number of stock s entering PWS
     Y,'t   =    number of stock s entering the spawning stream
     CM   =    catch of fish in district a

     p'ij   =    transition  probability  that a  fish  of  stock  s
               having left district i migrates to district j

     eM   =    probability stock s enters PWS through district a

     TM   =    residence time of stock s in district a

     q    =    catchability coefficient

     EM   =    fishing effort in district a


Movement of fish into and out of the district is  as follows:

                              immigration
             entry-
                                                     -catch
                        emigration & escapement

     N,at  =    entry  -  catch  -  emigration  +  immigration


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Entry is the number of fish entering the district from outside PWS
and is given by:
Catch is the number of fish removed by the  fishery and is known:



     C=a°E°yN
      *,t      "    M    " Mit
The catch can be apportioned to stock specific catch (  C'M ) by the
relative stock specific abundance:
     C'.
£   N.,.,.
stocks
Emigration is  the number of fish  migrating from the  district to
other districts or to the bay of the  spawning stream and is given
by:
             stocks
                                NM>t
Immigration is the number of fish migrating into the district from
other fishing districts and  is given  by:


           Z        Z  (  i  / TM )  NM>t  p\,t

        stocks   districts


Escapement is a component of emigration and  is given by:


          ( 1 / TM  )   N.t.>t   ( 1   -   I   P'.,t  )
                                170

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A  normal  probability distribution  timing function  f(*)  will  be
assumed for both the entry  (x,t) and for escapement  (y,t) :

     x,t   =    ffx-.XifM'i)


     yi>t   =    f(yaolla\fn\)


Where  x",, onlf p,\  are the total run,  standard deviation,  and mean
of the timing function for entry,  respectively; and y°°,, a'2,  M*2 are
the total run, standard deviation, and mean of the timing  function
for  escapement,  respectively.   Note that  the escapement  timing
function will be estimated directly from the  escapement data.

                        Fitting the Model

The  migration parameters  (  p'^,  e,a,  TM  )  will  be estimated  by
analysis of historical tagging data.  The method of estimation will
be based on Hilborn (1990) .  Both the forward  and  backward methods
of run reconstruction  (Schnute and Sibert 1983; Starr  and Hilborn
1988) with the forward method parameters of the model  (q, x00,,  a\,
H\ )  will  be estimated by  fitting  the model ( q S Nas )  to catch per
unit effort  (Ca / Ea) .   With the backward method, the  escapements
are lagged back to the districts based on a migration model derived
from the tagging data.
                            DISCUSSION

The life  history and run  reconstruction models will  accommodate
harvest in  existing mixed  stocks fisheries and  will enable  the
comparison of  alternative  commercial fisheries harvest  policies.
This  will  facilitate the  evaluation  of  fisheries  restoration
strategies that attempt to rebuild damaged stocks by reducing catch
in fisheries that exploit stocks damaged and stocks not damaged by
the oil spill.


                           BIBLIOGRAPHY

Hilborn, R.  1990. Determination of fish movement patterns from tag
     recoveries using maximum likelihood estimates.  Can. J.  Fish.
     Aquat. Sci. 47:635-643.

Schnute, J and J. Sibert.   1983.  The salmon terminal  fishery: a
     practical, comprehensive timing model.  Can.  J.  Fish.  Aquat.
     Sci.  40:835-853.

Starr, P.  and R. Hilborn.   1988.  Reconstruction  of harvest rates
     and  stock   contribution  in   gauntlet  salmon   fisheries:


                                171

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     Application to British Columbia and Washington sockeye salmon
     (Oncorhynchus nerka).   Can.  J.  Fish. Aquat.  Sci.   45:2216-
     2279.
                             BUDGET

Personnel                     $ 58.9
Travel                           5.2
Contractua1                    100.0
Supplies                         i.o
Equipment                       IQ.Q

Total                         $175.1
                               172

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FISH/SHELLFISH STUDY NUMBER 30

Study Title:   Data Base Management

Lead Agency:   ADF&G


                           INTRODUCTION

Large quantities of data are being analyzed in order to demonstrate
the extent  of injury  to  natural resources  due  to oiling.   The
purpose of this study is to make original data readily available in
electronic  form  to agency and non-agency personnel so  that data
analyses  can be  conducted in  an efficient and cost  effective
manner.  The data to be placed under the  database management system
(DBMS) will be drawn from two categories:

     1.   historical  data necessary  to  the interpretation  and
          implementation of the results of NRDA studies,
     2.   data resulting from NRDA studies.


                           OBJECTIVES

A.   To construct  a cost  effective DBMS to  readily  retrieve and
     order  data  from original  selected data in  electronic form
     according to user  specified criteria of time,  space, and other
     variables.     The DBMS  should  be  constructed  to meet  the
     following criteria,  in order of priority:

          1.   completeness of contents
          2.   speed of retrieval
          3.   ease  of  use  in  assembling  primary  data  into
               datasets for further analysis by other software.

     Furthermore,  the  DBMS will take advantage of  existing DBMS
     applications currently available in the ADF&G.

B.   To develop the structural facilities for individuals to access
     data  that is  physically located  at different  sites.   To
     accomplish this, a Local Area Network (LAN)  facility must be
     developed in  the  Cordova and Anchorage ADF&G offices,  along
     with a system for linking these with existing LANs in Juneau
     and Kodiak.   Note that Objective  B,  although  necessary for
     this project,  will  be  met  by  a   concurrent and  separately
     funded  "statewide database system" project  currently  being
     implemented by ADF&G using non-oil spill related funding.
                               173

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                             METHODS

A relational database management application will be developed.  It
will be based in standard structured analysis and structured design
methodologies.   Development will employ the industry standard SQL
language for relational databases.  The system will be accessible
by authorized IBM-compatible personal computers.  It will be made
available through  a linked  system of  LANs covering  offices in
Kodiak, Anchorage,  Cordova and Juneau.   The  end-user interface
software allowing non-programmer access to the database information
will be developed in Windows and made available to individuals.

The scope of data involves commercial species from PWS, Kodiak, CI,
and  Chignik  areas.     Specific   discussions   with  assessment
researchers  have prioritized  the  type  of observations  to  be
incorporated.  They are, in order of priority:

     1.   Commercial fisheries  catch  and  effort  data by  area,
          species,  and gear type.
     2.   Salmon escapement data,  including aerial survey counts,
          stream counts, weir counts, and sonar counts.
     3.   NRDA project data of global interest.
     4.   Preemergent and egg density counts.
     5.   Biological data  including age  composition,  size,  sex,
          growth, and stock composition.
     6.   Groundfish and shellfish survey data.

This project will make use of an ADF&G statewide database network
infrastructure being  separately  developed  with State  of  Alaska
general funds.   This project will not develop the network.
                              BUDGET

Personnel Services          $149.5
Travel                         5.4
Contractual                    7.8
Supplies                       2.6
Equipment                     10.5

Total                       $175.8
                               174

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               COASTAL HABITAT  -  INTERTIDAL  STUDIES

More than 1000 miles of coastal shoreline received light-to-heavy
oiling  from the  EVOS.   Assessment  of  injuries to  intertidal
resources and their rates of recovery require consideration of the
various categories of coastal morphology, the degree of oiling, the
specific   biotic    assemblages   affected,   and  their   trophic
interactions.  Assessment of clean-up effects is another component
of the injury assessment.

These  coastal  shorelines are  used by  many organisms which are
important to people, including  fish, shellfish, birds and mammals.
These  shorelines   are also  used  for  human  activities  such  as
recreation, fishing,  mining, and for  documenting past activities
through  invaluable  archaeological resources.    The  intertidal
studies are designed  to  estimate the  effects  of the spill and
associated clean-up  activities in  terms of:  (1)  the  abundance of
intertidal organisms and the  corresponding health of the ecosystem;
(2)   contamination  of   these  same   resources   by  oil;   (3)
quantification  of  injury from PWS to  the  KAP; and  (4)  natural
recovery of these resources.

These studies document the potential pathways of oil spilled in the
coastal environment as it moves through  the  food chain.  Thus, the
studies will provide data for determining  ecological  effects as
well  as  other  supporting data  for  determining  and  quantifying
injury to fish,  shellfish, mammals, and birds that provide services
directly to humans.  In addition,  these  studies serve as the basis
for  estimating rates of natural  recovery,  and the  need  and
potential for assisting natural recovery of the resources through
restoration.

Lastly, clean-up procedures  may not only reduce the adverse effects
of oil, but may also induce injury to intertidal resources.   The
assessment  of  clean-up effects by these  studies  is  an important
component of the overall injury assessment.
                               175

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COASTAL HABITAT INTERTIDAL STUDY NUMBER 1A

Study Title:  Comprehensive Assessment of Injury to Coastal
              Habitats

Lead Agency:  USFS


                           INTRODUCTION

The purpose of the Coastal Habitat Injury Assessment is to document
and  quantify  injuries  to biological   resources  found  in  the
intertidal zone throughout the  shoreline areas  affected by EVOS.
Field work in the supratidal zone was  concluded in  1990 and will
not be conducted in 1991, while the subtidal portion was integrated
into the formation of a 1991 suite of studies.

Study sites were selected and ground-truthed during Phase I.  Phase
II is an intensive evaluation of the study sites to determine the
extent of injury to natural resources.  The objective of this study
is to  estimate the  effects of  various  degrees  of oiling  on the
quantity (abundance and biomass), quality (reproductive condition
and growth  rate),  and  composition  (diversity  and proportion of
standing stock)  of key  species  in the  critical  trophic levels of
coastal communities.   These data are expected to provide evidence
of injury to the overall health  and productivity of these critical
coastal habitats, and  provide  information necessary to the more
species-specific  studies  on  the effects of  the oil spill  on
affected mammals,  birds and fish that use these habitats.
                             PHASE I

Selection and ground truthing of study sites were concluded during
1990.  No further Phase I work will be conducted during 1991.


                             PHASE  II

Injury Determination

Coastal habitats are unique areas of high productivity supporting
a diverse  array  of organisms, including  many  commercially and
ecologically important species.  These  habitats are particularly
vulnerable to oil spill impacts because  of the grounding of oil in
the  intertidal  zone,  the  persistence  of  oil  in  intertidal
sediments,  and the effects of associated clean-up activities.

Oil may affect coastal organisms directly by coating or ingestion,
with  toxic effects  leading  to  death  or reproductive  failure.
Indirectly, oiling may cause decreased productivity, accumulation
of toxic effects through the food chain, and loss of microhabitat

                               176

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such as  algae beds.   Assessment  of injuries to  coastal habitat
resources  and   determination  of  rates  of   recovery  require
consideration of the  various  coastal  geomorphologic  types,  the
degree  of  oiling,  the  affected  habitat,   and  their  trophic
interactions.   Ninety-seven  study  sites  comprised  of  59  sites
retained from 1989 and  38 sites added in 1990 were  selected for the
intertidal component  of  the Coastal  Habitat  Injury  Assessment
(CHIA).  These study sites are representative  of  the broad range of
coastal habitat  types including exposed rocky shores, fine textured
beaches,  coarse  textured  beaches,  sheltered  rocky   shores  and
sheltered estuarine  shores,  oiling  characteristics,  and clean-up
techniques found in the spill area.

Control sites were carefully paired with  oiled sites  to closely
match physical and biological characteristics while maintaining a
statistically valid site selection  strategy.   The current site
selection scheme will strengthen the ability of  the CHIA to detect
injuries while maintaining the ability to extrapolate these results
to the universe of other oiled shorelines.  From the original set
of 97 sites chosen in 1989-90,  a total  of 57 sites will be studied
in 1991.

Coastal intertidal animals may use  multiple habitats, necessitating
a  coordinated study of  the  effects of  oiling over  the  entire
intertidal  habitat.   The  complexity  of this  system  requires
expertise in  many disciplines.   Therefore,  an  interdisciplinary
team with the appropriate expertise,  including  plant  and systems
ecology,  marine  biology,   and statistical  analysis,  has  been
established.

The first year of field studies was  completed on November 1, 1989.
In 1990, field studies were conducted from approximately May 1 to
September 30.  In 1991, a May 1 to July 31 reduced field sampling
schedule is proposed.  Processing of samples and data analysis is
being conducted  to determine the variance and magnitude of changes
between unoiled and moderately and heavily oiled sites.
                            OBJECTIVES

A.   Estimate  the  quantity  (abundance  and dry  weight biomass),
     quality   (reproductive  condition   and   growth  rate),   and
     composition (diversity  and proportion of standing  crop)  of
     critical  trophic  levels  (and  subsequent impact  on trophic
     interactions)  in moderately and heavily oiled sites relative
     to unoiled sites.

B.   Estimate   hydrocarbon  concentrations   in   sediments   and
     biological samples.

C.   Establish the response of  these parameters to varying degrees
     of oiling and subsequent clean-up procedures.

                               177

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D.   Extrapolate impact results to the entire spill-affected area.

E.   Estimate the  rate of recovery  of  the habitats  studied and
     their potential for restoration.

F.   Provide  linkages  to other  studies  by  demonstrating  the
     relationships between oil, trophic level impacts, and higher
     organisms.


                             METHODS

Vertical transects will  be established  at 57 of  the  study sites
selected in Phase I.  Work will be conducted along  transects in the
intertidal zone.  For this study,  the intertidal extends from the
"0" tide  mark to  Mean Higher  High  Water (MHHW) .   Work  in the
supratidal zone was concluded in 1990.   Work in the subtidal zone
is being  conducted within the  context  of the  subtidal studies.
Community composition, cover,  and standing crop by trophic level
will  be estimated.   Key  species   (dominant  producers  and  food
sources) will be determined  and studied according to  the methods
listed below,  to estimate the quantity, quality,  and composition at
each trophic  level,  and to collect  samples  for determination of
hydrocarbon  contamination.     Using a  geographic  information
approach,  the impact  (by habitat type and  degree of oiling)  over
the entire area  affected by  the oil  spill  will  be integrated and
field-verified.

Specific methods for each component  of the  study were developed as
follows:

     Coastal
          1.  Initial Site Survey
          2.  Locating Transects
          3.  Sample Identification and Chain-of-Custody

     Intertidal
          Invertebrates
               1.  Locating 1 Quadrats
               2.  Swath Surveys
               3.  Reproductive Condition
               4.  Growth and Survivorship
               5.  Hydrocarbon Sampling Procedures
               6.  Experimental Work
               7.  General Laboratory Sorting Procedures
               8.  Subsampling of Intertidal Samples
               9.  Processing of Histological Samples
                               178

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          Fish
               1.  Locating Transects
               2.  Locating Quadrats
               3.  Sampling Quadrats
               4.  Minnow Trap Sampling
               5.  Sample Storage and Identification
               6.  Fish for Hydrocarbon Analysis


          Plants
               1.  Introduction
               2.  Study Plan
                    a.   Stratified Sampling
                    b.   Site Experiments at Selected
                         Habitats
                    c.   Field Experiments

Analysis of  samples  obtained in 1990 is  still  underway and will
continue as  additional  1991  samples  are collected.   Samples from
1991  will  be  processed as  rapidly  as  possible after  they are
returned from  the field.   The  reduced sampling scheme  in 1991
should allow for complete sorting  of 1990 and 1991 field samples
before commencement of any further field work.  The data from all
of the component studies are being entered into a computer database
management system.  This system is widely used,  and has good data
security features.   Use of  this database  system  will therefore
maximize both internal integration  and availability of the data to
related damage assessment projects.
                           BIBLIOGRAPHY

AOAC. 1980. Official Methods of Analysis  of the A.O.A.C., 13th ed.
     Chipperfield, P.N.J. 1953.  Observations on the breeding and
     settlement of Mytilus edulis (L.)  in British waters.  J. Mar.
     Biol. Ass. U.K. 32:449-476.

Johnson, R.D. and H.L.  Bergman.   1984.  Use of histopathology in
     aquatic toxicology:  A Critique. Pp. 19-36.  In V.W. Cairns,
     P.V.  Hodson  and J.O.  Nriagu,  eds., Containment  Effects on
     Fisheries,  John Wiley and Sons.

Ropes, J.W.  1968.   Reproductive cycle of the  surf clam, Spisula
     solidissima, in offshore New Jersey.  Biol. Bull. 135:349-365.

Seed,   R.   1969.      The   ecology  of  Mytilus   edulis   L.
     (Lamellibranchiata).  I.  Breeding and Settlement.  Oecologia.
     3:277-350.

Sheehan, D.C. and B.B.  Hrapchak.   1980.  Theory  and Practice of
     Histopathology. 2nd Ed. C.V. Mosby Co.
                               179

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Tietge, J.E., R.D. Johnson and H.L. Bergman.  1988.  Morphometric
     changes in gill secondary lamellae of brook trout (Salvelinus
     fontinalis)  after  long-term exposure to acid and aluminum.
     Can. J. Fish Aquat. Sci. 45: 1643-1648.

Tranter, D.J. 1958.   Reproduction in Australian pearl oysters. II.
     Pinctada albina  (Lamarck):   gametogenesis.   Aust.  J. Marc.
     Freshwtr. Res.  9: 144-158.

Wilson, B.R. and E.P.  Hodgkin.  1967.   A comparative account of the
     reproductive cycle of 5 species of marine mussels (Bivalvia:
     Mytilidae) in the vicinity of Freemantle, W. Australia.  Aust.
     J. Mar. Freshwtr. Res. 18: 175-203.
                              BUDGET

Services                  $
Travel
Contractual                 5,100.0*
Commodities
Equipment                   	

Total                      $5,100.0

*University of Alaska
                               180

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COASTAL HABITAT INTERTIDAL STUDY IB

Study Title:   Pre-spill   and   post-spill   concentrations   of
               hydrocarbons in sediments and mussels at intertidal
               sites within PWS and the Gulf of Alaska

Lead Agency:   NOAA


                           INTRODUCTION

Damage  assessment  of  the  oil  spill   in  PWS  and GOA  requires
information on hydrocarbon contamination levels  in water, sediment
and biota prior to the  spill  (baseline)  and  at various times after
the spill occurred, to  determine the potential impact and duration
of  impact.    Hydrocarbon baseline information  is available  for
several sites in PWS prior to oil transport  and  for the first four
years of  oil  shipment.  The  intertidal baseline  for hydrocarbon
levels in mussels,  sediment, water, and fish were established at 10
sites from 1977 to 1981. Ten additional sites were established in
the path  of the  spill in 1989.   All  sites  are located  on  low
energy, low gradient beaches,  often associated with eel grass. All
sites have adjacent bands of mussels (Mytilus trossulus).

Because of the potential persistence of hydrocarbons in sediments
in temperate and  subarctic intertidal  and subtidal environments,
sampling  will  be  continued  to document depuration  and recovery
rates.  Concentrations of  the full range of individual aliphatic
and aromatic hydrocarbons in sediments  and mussels from intertidal
sites will be reported.   Abundance of  mussels  and other epifauna
along  sediment  and mussel  transects  will be photographically
recorded during each sampling period.   These data will provide a
basis for estimating temporal and spatial impact to other biota of
the nearshore environment and support other NRDA studies of fish,
birds, and mammals.
                            OBJECTIVES

A.   To sample and estimate hydrocarbon concentrations in mussels
     and  sediments  from  20  sites within   10%  of  the  actual
     concentration   95%   of   the   time,   when   total   aromatic
     concentrations  are  greater than 200  ng/g dry wt.   We will
     compare these with 1989-90 data.

B.   To  test the  hypothesis  that  hydrocarbon contamination  of
     sediments and mussels is the same for  the pre-spill and post-
     spill period.

C.   To  document  changes   in  abundance   and  distribution  of
     intertidal  epifauna   and  test  the   hypothesis   that  no
     differences occur at oiled and unoiled sites.

                               181

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                             METHODS

Ten  intertidal  sites  in  PWS and  Port Valdez  were sampled  for
sediments, mussels, water, and fish annually from 1977  to 1981 to
establish a baseline against which  future  changes in hydrocarbon
concentrations can be  compared.  Sites  were initially  sampled in
spring, summer and  fall to determine if short-term changes occurred
during the warm season.  These sites were resampled in March 1989
immediately before several of them were impacted by the EVOS.

Immediately after the spill, and in some cases prior to the arrival
of oil, ten  additional sites were  established  to sample beaches
within the trajectory of the oil path.  Four of these sites were on
the KP  and  the  remaining six were  in  PWS.  Sediment  and mussel
samples were taken.  Photo-documentation was initiated along mussel
and sediment transects at each site.  These sites were re-sampled
several times during the summers of 1989 and 1990 to document the
appearance of and  changes in hydrocarbon  contamination  from  the
EVOS.   In 1991, only  the 16  sites in PWS will be sampled  and
sampling frequency will be reduced to once or twice during the warm
season.

Sediments;  Transect lines thirty meters (m)  in  length are located
parallel to the  water  line  at the -0.75 m  to +0.75  m  tide level
(depending on specific site).   Sediment samples are collected in
triplicate at each site.  Each sample consists of a composite of 10
cores  (dia 3.2 cm  x depth 1.25  cm)  taken  at random along the 30-
meter  transect.    Composite  sediment  samples  are  placed  in
chemically clean 4  oz.  jars, placed in an ice chest with artificial
ice  and  transported.    These  are  frozen  within  2-3  hours  of
collection.  One blank sample is taken at each site.

Mussels;  Transects for mussel collections  are located parallel to
the water line, usually immediately above the sediment transects at
approximately the +1 m tide level.   Triplicate mussel samples are
collected and each  sample  contains approximately 30 2-5 cm. mussels
(enough to produce >10 gms tissue)   taken at random along the 30-
meter transect.   Samples in 16 oz.  jars  are  cooled, transported and
frozen in the  same  manner  as the sediment samples.  All samples are
handled and stored according to established protocols to maintain
quality assurance and  control at all times.

Photo-Documentation;  Close-range views  of  the strata, macroflora,
and epifauna  are photographed.   Photos are  taken every  4 or  8 m
along the sediment transect  and every 2 or 4 m along  the mussel
transect line beginning at one meter.  Macrophyte  cover as well as
epifaunal occurrence  and density  are  recorded  from photographs
taken  of  625  cm2 quadrants  placed along the sediment  and mussel
transect lines.   A  grid of 100 random dots projected on each slide
                               182

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is  used to  estimate  occurrence and  percentage of  surface area
covered by macrophytes and epifauna.   Macrophytes and epifauna are
identified to species where possible.
                          DATA ANALYSIS

Random  sample and  subsample  collection prior  to  the  analysis
procedure  will ensure  that hydrocarbons  present in  the  sample
represent the average concentration at each site.  "Hot spots" of
hydrocarbon  concentration over  the 30  meter transects will be
cancelled out by this procedure.   Selected triplicate samples will
be  analyzed,  the mean  concentrations  and deviations  from these
means  determined,  and  appropriate statistical  tests  applied.
Digital tables of individual hydrocarbons will be reported.

Macrophyte and epifauna occurrence and cover will be analyzed using
one way ANOVA or paired comparisons  (oiled vs  unoiled where strata
are  similar).    They  will  be  tested  at  the  .05  level  of
significance.
                           BIBLIOGRAPHY

Connell, Joseph  H.   1970.  A predator-prey  system in the marine
     intertidal region.  1.  Balanus glandula  and several predatory
     species of Thais.  Ecol. Monog. 40:49-78.

Gundlach,  Erich  R.,  Paul D. Boehm,  Michel  Marchand,  Ronald M.
     Atlas, David M. Ward, and Douglas Wolfe.  1983.  The fate of
     Amoco Cadiz oil.  Science 221:122-129.

Karinen, John F., L.  Scott Ramos, Patty G.  Prohaska, and William D.
     MacLeod, Jr.  In Preparation.  Hydrocarbon distribution in the
     marine environment  of Port  Valdez and Prince William Sound,
     Alaska.

Krahn, M.M.,  C.A. Wigren,  R.W. Pearce, L.K. Moore,  R.G. Bogar, W.D.
     MacLeod,  Jr.,  S.Chan,   and  D.W.  Brown.  1988.    Standard
     analytical  procedures  of   the  NOAA  National  Analytical
     Facility,  1988.  New HPLC  cleanup  and  revised  extraction
     procedures   for  organic  contaminants.     NOAA  Technical
     Memorandum NMFS F/NWC-153.  52pp.

Warner,  J.  S.   1976.    Determination of  aliphatic and aromatic
     hydrocarbons in marine organisms.  Anal. Chem. 48:578-583.
                               183

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                              BUDGET

Labor                    $    31.0
Travel                        13.0
Contracts:  Helicopter        22.0
Supplies                       2.0
Equipment                      o.o

Total                    $    68.0
                               184

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Figure 1.   IntertI da I  baseline sampling sites.
         A  = historical sitesH  = established In 1989.
                                 185

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               SUBTIDAL RESOURCES INJURY ASSESSMENT

The  subtidal  regions of  PWS and  the GOA  represent  a vast  and
complex ecosystem.  The oil from the EVOS is known to have reached
portions of this ecosystem.  This subset of the NRDA studies have
the objectives of documenting the geographical extent, persistence,
and  toxicity  of the EVOS  oil  in  this environment  and examining
effects  of oil on  select  marine  organisms.   As the  natural
resources and  their  habitats in  the  subtidal  region  are  closely
related, the  studies on  them have  been placed together in a  new
Subtidal category for the 1991 NRDA study planning process.  This
category  of  studies  includes  the  former  Air/Water  studies,
including studies of benthic infaunal communities,  and studies of
various species of demersal fish  and shellfish.

Water Resources

Monitoring of the concentrations  of petroleum hydrocarbons in the
water column of PWS and portions of  the GOA began immediately after
the EVOS.  This monitoring was most critical during the first few
weeks  following  the  spill  when  the  dissolution  of  soluble
components was most rapid and the likelihood of toxic exposure was
highest.  As dilution of the EVOS  oil in the water column continued
below the levels  that can be detected using direct measurements,
the  strategy  for  long-term documentation  of  the  locations  and
concentrations  of  hydrocarbons  available  to  marine  organisms
shifted to the use of alternate means  of detection.  This involved
the study of bioaccumulators and measurements of the  settling rates
of oil contaminated sediments settling out from the water column.
Subtidal Study No. 3  is dedicated to carrying out this monitoring.

Marine water  quality is  protected under state and  federal water
quality standards which  include classifications  for such  uses as
growth and propagation of  fish and wildlife,  aquaculture, and human
uses such as recreation.   Moreover, State of Alaska water quality
standards for petroleum hydrocarbons establish criteria for water
habitats.

Sediment Resources

A portion of the EVOS oil reached the marine sediments in PWS and
in portions  of the GOA.    The  extent of this  contamination,  its
persistence and toxicity, and its  direct  effect on  the  benthic
communities living in contact with sediments are studied by three
of  the studies  in this  category.    Subtidal Study  No.   1 will
investigate the occurrence, persistence, and chemical composition
of petroleum hydrocarbons in marine  sediments.  Subtidal Study No.
2 will document the effects of EVOS oil in marine  sediments  on deep
and shallow water benthic communities.  Subtidal Study No. 4 will
investigate  the  fate  of  EVOS oil and  determine  its long-term
toxicity.


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These studies will document the injury level to a large ecosystem
which contains a  large  number of organisms that are  in the food
chain of many higher trophic level animals that are the subject of
other NRDA studies.

Demersal Fish and Shellfish Resources

Subtidal studies 5, 6, and 7 have the goal of documenting exposure
to EVOS oil and injury for a number of demersal fish and shellfish
resources.  These studies combine elements of 1990 Fish/Shellfish
studies 15, 17, 18, and 24.  The large number of demersal species
potentially  affected by  the  EVOS  and  the vast  extent of  the
available  habitat that  they occupy has  resulted  in  these 1991
studies being primarily  focused on representative species in areas
of  PWS  where  the potential  for injury  is believed  to be  the
greatest.

The  demersal  fish/shellfish  resources  of  PWS and  the GOA,  in
addition to being utilized  by commercial,  sport,  and subsistence
fishermen, are a  key food source for  other fish,  marine mammals,
river otters, and for various species of birds.
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SUBTIDAL STUDY NUMBER 1

Study Title:   Hydrocarbon  Exposure,  Microbial  and  Meiofaunal
               Community Effects

Lead Agency:   NOAA, DEC


                           INTRODUCTION

A substantial proportion of the approximately 11 million gallons of
Prudhoe  Bay  crude  oil  released  into  the  marine  environment
following the grounding of the tanker Exxon Valdez became stranded
on the shoreline of PWS  and northeastern GOA.  Some of the oil that
entered the water  (the original crude oil derived from the spill,
oil leaching  from contaminated shorelines, and/or  oil  dispersed
into  the water  by shoreline  cleanup  activities)  reached  the
subtidal region as  a result  of  physical  and biological  processes
(Boehm et al.  1987).    The proportion  of the original  volume of
crude oil spilled from the Exxon Valdez that has reached subtidal
sediments in PWS remains to be determined.

NOAA

A primary objective of  the present study  is to synthesize the data
on hydrocarbon contamination of subtidal sediments collected by all
NRDA studies.  This will allow an estimate of the amount of crude
oil that contaminated subtidal  sediments  in  PWS and GOA and define
the geographic and bathymetric extent  of subtidal  hydrocarbon
contamination.  Sampling of subtidal sediments in PWS will continue
on a reduced scale in order to resolve the dynamics of hydrocarbon
contamination  of  subtidal  sediments  influenced  by  additional
contamination  resulting  from  1990  cleanup  activities and  the
persistence of  petroleum  hydrocarbons  in previously contaminated
sediments.

DEC

The DEC portion of this study will conduct microbiological assays
to measure the response of microbial  populations  to the  EVOS.  The
intertidal  and  subtidal sediments  for this portion  of  the study
will be collected at the same sites where the NOAA sediment samples
are taken.

Assessment of microbial  populations is important  since the ultimate
fate of spilled oil depends on the ability of microorganisms to use
it as a source of carbon and  energy  (Leahy and Colwell 1990).  The
microbial hydrocarbon oxidation potential  assays are designed to
measure microbial activity under optimized environmental conditions
and independent of "in situ" hydrocarbon  concentrations.   Thus,
they are an indicator of the microbial communities' acclimation to
particular  hydrocarbon  fractions,   implying  exposure  to  these

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petroleum  components "in  situ."    The observation  of microbial
communities acclimated to hydrocarbon oxidation in intertidal and
subtidal  sediments  only  implies  exposure  to  hydrocarbons  in
general.    Definitive characterization  of  the  hydrocarbons  as
originating from the Exxon Valdez  will  depend on detailed chemical
analysis  of the  sediment  samples  collected in  parallel  to the
microbiological samples.

The  sediment  sampling will  be coordinated  closely  with benthic
infaunal  studies  (Subtidal Study  No.  2) .  The  benthic study will
examine the effects of the oil spill on infaunal communities below
a depth of  20  m.   The sampling for this  study will be conducted
from  the  same vessel  simultaneously  (June/July  1991)  as  the
deepwater sediment  sampling.   The  second  study will examine the
effects of the  oil on infaunal communities  associated with eelgrass
and Laminaria beds.  Sediment and microbiological samples will be
collected at the same sites where infauna of the eelgrass community
will be taken.   The benthic infaunal studies will be described in
detail  in a separate  plan.    The sediment  and benthic infaunal
studies were combined in the Air/Water Study No.  2 in 1990.
                            OBJECTIVES

A.   Synthesize the  analytical results on  the  concentrations of
     petroleum hydrocarbons in subtidal marine sediments collected
     under  this study  and all  other NRDA studies  under  which
     sediments have been collected.

B.   Determine occurrence,  persistence, and chemical composition of
     petroleum  hydrocarbons  in  all   subtidal  marine  sediments
     analyzed to date.

C.   Provide  marine  sediment  data to generate in  mass  balance
     calculations on the fate of oil in the marine environment.

D.   Enumerate  populations of hydrocarbon-oxidizing  microbes in
     intertidal  and  subtidal  sediments  collected at  oiled  and
     unoiled sites within PWS.

E.   Assess the maximum potential for "in situ" biodegradation of
     selected hydrocarbon substrates in subtidal sediments at oiled
     and unoiled sites within PWS.
                             METHODS

NOAA

Sediments will be sampled  at  20  sites in PWS (10 reference sites
and 10  contaminated sites).   Fourteen sites will  be  sampled in
June/July.    Sediment  sampling   will  be  coordinated  with  the

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microbiological and  deep  benthos projects at  these 14 sites.
Nine sites will be sampled in May and September.

Three  samples,  each  a  composite of  eight subsamples  collected
randomly along 30 m transects  laid parallel to the shoreline, will
be taken at each intertidal site.  Samples will  be collected at low
tide or by divers.  Intertidal collections will  be made at a single
tidal height in the range  of +1  to -1 m relative to mean lower low
water  (MLLW) depending on the distribution of fine sediments.

Subtidal sediment collections  will be made at 6  m below MLLW in May
and  September and  at  3,  6,  20,   40  and  100 m   in  June/July.
Collections at 3, 6  and 20 m  will be made by divers on transects
laid along the appropriate isobath and sampled  in the same way as
described  above  for the  intertidal  transects.    The  eelgrass
community project will sample sediments,  infauna  and epifauna in
the same depth range at  six  of the PWS  sites.   Samples  taken at
depths below  20 m will  be collected with  a  Smith-Mclntyre grab.
Three grabs will be taken at each depth.   Four  subsamples will be
removed  at randomly selected  points within  each grab.    The
subsamples  will  be combined  to form one  sample  per grab.   The
samples will be taken at the  same sites  as the benthos (see deep
benthos  sampling in  the  Subtidal  Study  No.  2  plan),  however
sediments  will  not be taken  from the same  grab as the  benthos
samples  because  the  volume  needed  for  sediment  hydrocarbon
analysis.

DEC

Sediment samples for  the microbiological work will be obtained from
sediment chemistry samples taken during the June  cruise.   Samples
will be taken at all  14  sites  and at all depths where the sediment
chemistry samples are taken.   The samples taken by divers at the 3
m, 6 m and 20 m depths will be generated by placing approximately
1 kg of surface sediment in sterile whirlpack bags,  and sealed at
the sampling depth.   The 40 m and 100 m  samples  will  be obtained by
composite subsampling into a sterile whirlpack bag of the surface
sediment  contained  in  the   sampling  device.    The  intertidal
microbiological  samples  are  composites   of  eight  subsamples
collected at random intervals  along a 30 m transect parallel to the
shoreline  in  the  low intertidal zone.    All microbiology samples
will  be collected  as triplicate  composites  from   the  transect
sampled.

Care  will  be  taken   to  avoid  contamination   of  samples  by the
sampling  personnel  and  cross-contamination  between  different
sediment samples.   Sampling apparatus will be thoroughly rinsed
with  water  between   samples  and  disinfected with  alcohol  or
alternate disinfectant.   Samples obtained from  the deepwater grabs
will be collected from the center of the core to avoid surface
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contamination incidental to sample handling.  All microbiological
samples will  be placed in  coolers  for transport  to  the support
vessel for processing within three hours of collection.

Hydrocarbon  biodegradation  potential  associated  with  sediment
microbes will  be assayed by adding  radiolabelled  aliphatic  (14C-
hexadecane) and aromatic (14C-phenanthrene) substrates to sediment
samples.   Each  substrate will  be monitored for biodegradation by
the evolution  of  radio-C02 from  the  samples after  two incubation
periods.

A total of 20  gm  of  sediment from each sample will be needed for
this  assay.    Each  sediment  sample  assayed  for  hydrocarbon
biodegradation  will  first  be  mixed  1:10 with  sterile  seawater
augmented  with  mineral nutrients   (Difco marine  Bushnell-Haas
broth) .   Ten ml  aliquots  of the  resulting slurry will  then be
placed  in  sterile 40  ml incubation  vials fitted  with  silicone
septa.  The substrate of interest will be  added at  a 10 ppm  (ug/ml
slurry) concentration by injection via syringe through the septa.
The substrates will then be added in an acetone carrier (Baur and
Capone  1988).   Two  replicate  vials  for  each substrate/sediment
sample/incubation time  combination will be prepared with a "time
zero"  killed  control  also  prepared  for  each  substrate  and
triplicate set.  AH  vials will be placed on a rotary shaker  for 24
hours and then incubated at ambient temperatures for the duration
of the incubation period.

Following incubation of the sample for the appropriate period (or
initially in the case of the controls), substrate biodegradation in
the sample vials will be halted  by the addition  of 1 ml ION NaOH
through the  septum.   This will  result in a pH  greater  than 13,
killing the culture  of  degraders  and  sequestering  any  evolved C02
in the form of carbonates in solution.  The extent of hydrocarbon
degradation will  be  monitored  by measuring the radio-CO2 evolved
from  each  vial  (Foght et  al.   1988) .    After  transport to the
analytical facility  at  the  University of  Alaska,  the sample vial
contents will be purged of radio-C02 and the effluent gas will be
passed  first  through  an  organic vapor  trap  and then  through
phenethylamine  scintillation cocktail  to  trap  the  evolved C02
(Fedorak et  al.  1982).  The mean of each  set  of  biodegradation
samples for  each substrate, concentration  and  incubation period
will  be  compared the "time  zero" killed controls  to  assess for
losses due to volatization in transit or  any  possible  abiotic CO2
evolution.   The extent of biodegradation will be  expressed as a
percentage of the total radiocarbon  activity added to the sample
after correction for abiotic losses.

In addition to the biooxidation potential assay,  all microbiology
samples will  be  analyzed  using  the  Sheen Screen  Most  Probable
Number  technique  for  the  presence  of  surfactant  producing,
hydrocarbon-degrading microorganisms  (Brown and Braddock 1990).


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While  no  technique  to  enumerate  specific metabolic  types  of
microorganisms  in  marine systems  is  absolute,  the  Sheen Screen
technique  provides  consistent results  that are  appropriate  for
relative comparisons among stations and depths.


                          DATA ANALYSIS

NOAA

Synthesis of Sediment Analyses

Sediment samples  collected  for 12  studies  included in  the NRDA
process have been catalogued in the damage assessment database of
Technical  Services  Study No.  1  and some  of those  samples were
submitted for analyses.  The principal goal of the present proposal
will be  to synthesize  the results  from  the sediment hydrocarbon
analysis as they become available from Technical  Services Study No.
1.   Mapping  of the geographic and bathymetric  distribution  of
hydrocarbon contamination of sediments  in PWS and the northeastern
GOA will be carried out in coordination with the  DNR.  The combined
sediment data will also be used to test specific hypotheses about
the distribution of Exxon Valdez  oil  in sediments throughout the
study area.

Statistical Analysis

In general, for sediment analyses the null hypothesis states that
the concentration of petroleum hydrocarbons at particular depths or
the distribution  of petroleum hydrocarbons with depth  at oiled
sites does not differ from that at reference sites.  All data will
be  tested  for  heteroscedasticity  with   Bartlett's   test  or
equivalent.   Data will be  reported as means and 95%  confidence
intervals  calculated according to a standard formula  (Sokal and
Rohlf 1981).  Parametric statistics (Model I analysis of variance
with site  and depth as fixed factors  and Scheffe's a  posteriori
test)   will  be  used  to test for differences  in  hydrocarbon
concentrations between sites and depths if underlying assumptions
of the parametric procedures are met (with data transformation if
required),   otherwise nonparametric tests (eg.  the Kruskal-Wallis
test)  will  be employed.  Key petroleum weathering and source ratios
will be calculated (Boehm et al.   1987).
Data on microbial activity levels  and hydrocarbon degrader numbers
will be subjected to non-parametric analyses (e.g. Mann-Whitney U
test) to  demonstrate any significant  statistical  differences in
microbial community responses at oiled and reference sites.
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                           BIBLIOGRAPHY

Bauer,  J.E.  and  D.G.  Capone.    1988   Effects  of co-occurring
     aromatic hydrocarbons  on degradation of polycyclic aromatic
     hydrocarbons  in  marine  sediment slurries.    Appl.    Env.
     Microbiol.  54:1644-1655.

Boehm,  P.  D.,  M.  S.  Steinhauer,  D.  R.  Green,  B.  Fowler,  B.
     Humphrey, D.  L.  Fiest and W.  J.  Cretney.  1987.  Comparative
     fate  of  chemically  dispersed  and  beached  crude   oil  in
     subtidal sediments of the arctic nearshore.   Arctic 40, supp.
     1: 133-148.

Brown,  E.J.  and  J.F.  Braddock.     1990.    Scheen  Screen,  a
     miniaturized  most-probable-number method for enumeration of
     oil-degrading microorganisms.  Appl.  Env. Microbiol. 56(12):

Fedorak, P.M., J.M. Foght and D.W.S. Westlake.  1982. A method for
     monitoring mineralization of 14C-labeled  compounds in  aqueous
     samples.  Water  Res.   16:1285-1290.

Foght, J.M., D.L.  Gutnick  and D.W.S.  Westlake.   1989.   Effect of
     Emulsan  on  biodegradation of  crude  oil by pure and mixed
     bacterial cultures.  Appl. Env. Microbial. 55:36-42.

Gundlach, E.  R.,  P. D. Boehm,  M. Marchand, R.  M.  Atlas,  D. M. Ward,
     D. A. Wolfe.   1983.   The fate of Amoco Cadiz oil.    Science
     221:122-130.

Leahy, J.G.  and R.R.  Colwell.   1990.   Microbial degradation of
     hydrocarbons  in  the environment.  Microbial. Rev. 54(3):SOS-
     SIS.

Sokal, R. R.  and F. J. Rohlf. 1981.  Biometry.  W. H. Freeman and
     Company, San  Francisco.  859 pp.
Salaries
Travel
Contracts
Supplies
Equipment
Vessel

Total
NOAA

$123.0
  18.0
  20.0
   6.0
   8.0
 120.0

$295.0
 BUDGET

  DEC

 $28.0
   3.5
 107.5
   0.8
   0.0
   0.0

$139.8
 Totals

$151.0
  21.5
 127.5
   6.8
   8.0
 120.0

$434.8
                               193

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SUBTIDAL STUDY NUMBER 2

Study Title:   Injury to Benthic Communities

Lead Agency:   ADF&G

Cooperating Agencies:   DEC and NOAA


                           INTRODUCTION

Benthic  organisms   (both  meiofauna  and  infaunal  macrofauna)
associated with subtidal sediments generally represent good in situ
monitors for measuring effects of  oil fluxing to the bottom (for
example see Cabioch et al.  1978; Kineman et al. 1980; and Sanders
et al. 1980).   These organisms typically remain close  to or at the
site  of larval  settlement,  and,   consequently,  represent  good
monitoring organisms.  The  composition of  the marine benthic fauna
has been  successfully used  at  various  locations throughout the
industrial world as a basis for measuring  effects of pollutants on
the bottom  (e.g., see  Pearson 1975; Cabioch et al.  1978; Pearson
and Rosenberg  1978; Gray  and  Mirza 1979;  Sanders  et  al.  1980;
Kineman et  al. 1980; Gray  and Pearson 1982; Warwick 1986;  Boesch
and Rabalais 1987;  Warwick  et al. 1987; and  Gray 1989), and should
prove useful for assessing biological effects of the EVOS in PWS.

Subsequent to the crude oil  spill  from the  EVOS,  it was expected
that a  certain proportion  of oil in the  water column (either the
original  crude  oil derived  from  the  spill,  oil   leached  from
contaminated shorelines, and/or oil dispersed into receiving waters
via shoreline  remediation  procedures)  would reach  the  bottom by
physical and  biological processes.   Benthic data  collected  in
polluted waters elsewhere indicate  that changes in species number,
abundance,   biomass,  and diversity  occur  if  sizable  quantities of
oil flux to the  bottom.  Changes in composition of  benthic fauna
can have serious trophic implications since many subtidal benthic
invertebrates  are   important food  resources  for  bottom-feeding
species such as  pandalid shrimps,  crabs,  bottomfishes,  sea ducks
and sea otters  (see review in Feder and  Jewett 1981, 1987; Hogan
and Irons 1988; McRoy 1988).  Further,  the  larvae of most benthic
organisms in PWS move  into the  water  column (March  through June)
and are utilized as  food  by large  zooplankters and larval and
juvenile stages of pelagic fishes,  small  salmon fry,  and herring.
Thus,  damage to the benthic system by  hydrocarbon  contamination
could affect  feeding  interactions of important species  on the
bottom as well as in the water column.

Shallow (<20 m) subtidal studies were initiated in PWS in the fall
of 1989 and continued during the summer of  1990 under the Coastal
Habitat Study.   Deep (>20 m)  benthos studies were initiated in PWS
in July 1990 under Air/Water Study  2 (Injury to Deep Water  [>20 m]
Benthic Infaunal Resources  from Petroleum Hydrocarbons) .  Six of the

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deep benthos sites sampled in 1990 were adjacent to eelgrass sites
sampled by the shallow benthic program.

Sampling (for at least five years) of subtidal benthic populations
should be continued as a method for assessing possible effects of
oil on benthic  communities as related to  redistribution  of oil-
laden sediments from adjacent contaminated onshore sites.  Oil that
initially coated  sediments onshore  may  eventually be transported
offshore,  thereby  contributing to long-term  effects  on  deep
subtidal benthic  fauna.  Examples of  such effects  were  observed
following the Amoco Cadiz  crude oil spill  of 1978,  in the Bay of
Morlaix off the Brittany  coast of France  (Cabioch  et al. 1978) and
following the Florida No. 2 fuel oil spill  of 1969 in Buzzards Bay
near West Falmouth, Massachusetts (Sanders et al. 1980).
                            OBJECTIVES

Shallow Benthos

A.   Determine the temporal and spatial effects  of the EVOS on the
     infaunal   invertebrate  communities  within   selected   PWS
     embayments  where  eelgrass  (Zostera)  and  the brown  algae
     (Laminaria) dominate.

Deep Benthos

A.   Determine  if  changes occurred in  the benthos  following the
     EVOS by comparing  taxon  (primarily determined  at the family
     level: see Methods) richness and diversity, general abundance
     and biomass, and trophic composition of benthic biota living
     on similar substrata at stations at depths of approximately 40
     and 100 m below eelgrass beds in oiled and unoiled bays.

B.   Determine  if  changes occurred in  the benthos,  as estimated
     temporally, by comparing taxon (see objective above) richness
     and diversity, general abundance and biomass, and the trophic
     composition of  benthic biota  at  stations within  oiled and
     unoiled bays on an annual basis for at least five years.

C.   If changes are detected  in the  infaunal  components  of the
     benthic system, determine  how much time  is required for the
     benthos to recover to a relatively stable assemblage of taxa.

D.   If  changes   are   detected  in the  infauna,   examine  the
     relationship  between  the  accumulation  and  retention  of
     hydrocarbons in sediments and the  effect  on the benthic biota
     (this will be accomplished in conjunction with the subtidal
     project assessing   hydrocarbon levels  in sediments  at the
     sampled stations).
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                             METHODS

Shallow Benthos

General

Shallow subtidal  sampling efforts  will concentrate  on  infaunal
invertebrate communities in eelgrass and Laminaria habitats within
bays in PWS.  These habitats were also sampled in 1990. They were
chosen based on their relative ecological importance, the history
of prior damage, and on  their proportion  of  total  habitat in the
oiled area.  Six of the sites within the eelgrass habitat were in
common with the Deep Benthos sites. All studies will be conducted
at oiled sites  (selected  at random when possible) and control sites
that are matched to the oiled sites with regard to geomorphology,
degree of  freshwater input, substrate type, and general circulation
and wave exposure regimes.

The shallow subtidal sampling for 1991 will occur in concert with
the rockfish studies to be conducted by ADF&G (Subtidal Study 6).
Both studies will utilize the same divers on the same platform to
sample the  shallow waters in western PWS.  Some of  the  sampling
sites for the two studies are in common.

Stratified Sampling - Rationale

A stratified sampling design, modified from the design used in our
1990 survey, will be employed in order to obtain estimates of basic
population   parameters    (density   and  biomass)   for   infaunal
invertebrates.    These  estimates will  be used  to  indicate  the
effects of  the  EVOS  on this community  by comparing  density (and
other parameters)  at oiled vs.  control sites.  The data will also
be used in support  of  other studies  (e.g., otters and birds) since
the animals within  the subtidal habitats are major food sources for
these other species.

Strata to be sampled

In the 1990 sampling,  the shallow subtidal communities within PWS
was stratified  into three major habitat types  based  on the dominant
plants within the  habitat: Nereocystis beds, eelgrass  beds,  and
Laminaria beds  (areas where either Laminaria saccharina or Agarum
cribrosum dominate).  For the Laminaria habitat (the most widely
distributed), we further stratify  into 3  oceanographic regions:
islands, mainland,  and  outer sound and into three physiographic
types:   bays,  points,   and  runs  (straight  shore  line).   This
stratification  scheme resulted  in 9  potential  strata within the
Laminaria habitat,  1 within the Nereocystis habitat, and 1 within
the eelgrass habitat, for a total of  11 potential  strata in all.
Another  strata,  silled  fjords,  was  added in   1990  based  on
preliminary finding from  our 1989 survey.


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In 1990, we  sampled 5 of the 12 potential  strata:   Nereocystis,
eelgrass, Laminaria in island bays, Laminaria on island points and
silled fjords.  In  1991, we  will sample only  in the eelgrass and
Laminaria bay habitats.

Selection of sites within strata

Sites to be sampled in 1991 are  a subset of those visited in 1990.
These were  selected based on the  summer 1989  oil maps  and the
September 1989  "walkathon" data.   Areas that were  moderately to
heavily oiled in both  surveys will be  used  as  oiled sites.  From
these oiled areas for  each strata  (i.e.,  island bays or eelgrass
beds), a section of shore  line was selected to be  sampled.   The
selection of  the  sampling locations was based on  the following
hierarchy for order  of  preference:  sites for which there were pre-
spill biological data, sites previously  sampled  in NMFS  or DEC
hydrocarbon surveys, sites sampled by  Coastal  Habitat intertidal
crews,  randomly selected  sites within the  habitat,  and sites
sampled in the deep benthos study.

Control sites were  selected  that were  unoiled  in  both the summer
oil  survey and  the  "walkathon."    Controls  were  matched  with
selected oiled sites with regard to aspect, proximity to sources of
freshwater input,  slope, wave exposure, and water circulation.  A
matched site will be selected randomly if more than one exists.

Initial site selections were  made based on  oiling maps and input
from scientists familiar with habitats  within PWS, as well  as from
fishermen familiar  with PWS.   Final  selections  were made  in a
reconnaissance survey conducted in April, 1990.

A total  of 3  to  5  oiled  sites and  3 to  5  control  sites  were
selected from  each habitat.   Three of the oiled/control pairs
within the eelgrass habitat  are also  sites for the Deep Benthos
Component.  In 1990, shallow and deep benthic sampling occurred at
the following oil/control paired sites: Bay of Isles (O)/Drier Bay
(C) ;  Herring  Bay   (O)/Lower  Herring  Bay  (C) ;  and  Sleepy  Bay
(0)/Moose Lips Bay  (C).

Data Analysis

All taxonomic  identifications for  the  1991 sampling  period will
only be taken to the  family  level  to accelerate processing time.
Data analysis will be coordinated with analyses performed under the
Deep Benthos component.

The  general  form of  analysis for  all data gathered will  be a
comparison of  oiled vs.  control  sites  using  t-tests  or nested
analyses of  variance.    In  studies where more than one  site is
sampled, sites  will be the  primary  sampling unit,  with various
degrees of subsampling within a site.
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Deep Benthos

Sampling

The sampling  plan for the project  calls for collection  of five
replicate  samples at  each  of  two  stations within seven  bays
identified as oil-exposed sites  and two stations within seven bays
determined  to have  been uncontaminated (control)  sites.    All
stations sampled will be at approximate depths of 40 and 100 m on
a  transect extending  below  seagrass  beds  within  each  of  the
identified bays.   Shallow subtidal stations on the transects for at
least eight of the bays will be sampled for biota for the Shallow
Benthic Studies.   A total of 28 deep stations x 5 replicates will
be collected on a single  cruise in  July  1991 in conjunction with
microbiological and  hydrocarbon sampling  projects that  will  be
underway from  the same ship platform.    Shallow subtidal  benthos
(<20 m)  will be sampled at approximately  the  same time period from
a different ship platform, a circumstance necessitated by the need
for a special ship-diving platform.   Deep benthic samples at oil-
exposed and unexposed sites will be collected on bottoms that are
as physically similar as possible.  The seven oil-exposed sites to
be sampled for deep benthos are Northwest  Bay,  Disk Island, Herring
Bay, Bay of Isles, Snug Harbor, Sleepy Bay, and Chenega.  The seven
unexposed  (control)  sites to be sampled for deep benthos are West
Bay, Rocky Bay, Zaikof  Bay,  MacLeod  Harbor,  Mooselips  Bay,  Lower
Herring Bay, and Drier Bay.

Deep benthic biological samples at stations at approximately 40 and
100 m will be  collected with a  0.1  m2van Veen grab weighted with
31.7 kg of lead to facilitate penetration.  Five replicate samples
will be taken  at  all stations.   Material from  each grab  will be
washed  on nested  1.0  and  0.5 mm  stainless  steel screens  and
preserved in 10% formalin-seawater solution buffered with hexamine.

Analysis and Processing Data

Organisms that will be collected by grab and subsequently used in
analyses  include  infaunal   macrofauna,   slow-moving  macrofaunal
surface  dwellers, and  small  sessile  epifauna.    Highly motile
epifauna such  as  large  gastropods,  shrimps,  crabs, and sea stars
(except  the infaunal  sea star, Ctenodiscus crispatus)  are  not
adequately  collected by grab and  will  not  be  analyzed.   Since
0.5 mm  mesh   fractions  were   collected   and   sorted,   larger
representatives of the meiofauna that are retained  quantitatively
by this screen will be analyzed.  Thus,  the following organisms are
included  in  the analyses:   nematodes,   tardigrades,  ostracods,
harpacticoid   copepods,  tanaids   and   cumaceans.     Although
Foraminifera were common at  some stations, most  specimens examined
were dead  at  the time of collection.  Additionally, the sorting
time necessary to sort  samples  required  that (up to 60 hours per
0.5 mm replicate) this group  be deleted  from the analyses.  Thus,


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Foraminifera are not included in the analyses;  however, all samples
containing large numbers of Foraminifera are archived.

All  organisms  will  be  identified  primarily  to  Family  or  an
appropriate  higher   taxonomic   level.     Generic  and  specific
determinations for organism will be made whenever these categories
are known and will  be recorded on the data sheets.  The decision to
use higher taxonomic categories is expected to increase the speed
of  processing  samples.    Earlier  analyses  of  benthic  samples
obtained  at  study  sites  shortly  after  the EVOS  indicated  that
species diversity  was  typically  high.   It was estimated that the
time  necessary  to  determine taxa  to generic and  species  levels
would result in a multifold increase  in hours  spent  in sorting and
taxonomic identifications.  Additionally, a recent paper by Warwick
(1988) and other papers (Rosenberg  1972; Heip et al.  1988) indicate
that better resolution of multivariate and other data emerges when
higher taxonomic levels are used.  However, availability of generic
and specific names for common  organisms  allows  an examination of
station data in more detail if any of these taxa are particularly
abundant at  a  site.   All  individuals are counted and weighed by
taxonomic group.   Approximate carbon values  for  all wet-weights
will be calculated.

All data will be recorded on data sheets,  entered on magnetic tape
and processed with the VAX computer at  the University  of  Alaska
Fairbanks. Previously written programs at the  University of Alaska
for comparison of rank abundance and biomass will be  applied to the
PWS  data.    A diversity program  will  also  be  used to  examine
differences and similarities between stations.

Numerical Analysis

Station groups  and taxon assemblages for  each year,  and  for the
combined data collected on  subsequent cruises in  future years, will
be identified using the technique of hierarchical cluster analysis.
Principal  coordinate  analysis  will  be  used as an  aid in  the
interpretation of the cluster  analysis of the  data and to identify
the misclassification of stations by cluster analysis. Use of both
of these multivariate techniques will make  it possible to examine
similarities (or dissimilarities)  between groups of stations, and
should be useful when comparing oiled vs unoiled bays.

A Kruskal-Wallis and a multiple comparison test for significance
will be used to test for  differences in  the  total abundance and
biomass between stations sampled each year  and in multi-year data
sets.  These tests will be made on  the  abundance and biomass of
selected, dominant taxa  at stations between  years.  Taxa  will be
chosen from  the rank abundance and  biomass  printouts  for  each
station;  taxa  selected will generally be those  commonly present
within bays  being  compared.   However,  taxa  that  are  common at
stations within unoiled bays,  but rare  or missing  at  stations
within oiled bays,  will also  be  tested.   Analysis  of  variance

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(ANOVA) will  also be used  to  test differences in  abundance and
biomass between dominant taxa for stations at similar depths within
unoiled and oiled bays.

Various measures  of diversity will  be calculated,  and compared
between stations at similar depths within unoiled and oiled bays.
The indices to be calculated and presented are: Shannon Diversity
(measures  total   diversity),   Simpson   Dominance   (useful  for
identifying  dominance  by  one or    a  few  taxa  at a  station) ,
Evenness,  and Species Richness.

The K-dominance curves  (Warwick  1986) that relate  abundance and
biomass data  will  be  used in an attempt  to assess  the  effect of
hydrocarbons on benthic organisms in  oiled bays.  This is a recent
technique  designed to  detect pollution-induced  disturbance  on
marine  benthic communities.    However,  there are  problems  of
interpretation  of  the  output of  this  technique  that must  be
considered before environmentally-related conclusions can be drawn
(Gray 1989; Beukema, In press). Distributions of geometric classes
of abundance of species will also be calculated (Gray and Pearson
1982).  Assessment  of the distribution of taxa in these abundance
classes is  often  useful to identify indicator species  within a
disturbed area.

Methodologies, rationale, and  problems  with the  use of diversity
indices, K-dominance  curves,  and geometric abundance  classes as
measures of pollution-induced disturbance are discussed in Bayne et
al. (1988), Gray et al. (1988) and Appendix C.


                          BIBLIOGRAPHY

Bayne, B.L.,  K.R.  Clarke, and J.S. Gray.  1988.  Biological Effects
     of Pollutants.  Results of a Practical  Workshop.  Mar.  Ecol.
     Prog.  Ser.  46.   MEPS  Special Book version.  Inter Research,
     Federal Republic of Germany.  278pp.

Bartlett,  M.S.  1936.  The  square root transformation in analysis
     of  variance.    Journal  of  the  Royal Statistical  Society
     Supplement 3:  68-78.

Beukema,  J.J.    An  evaluation  of  Warwick's  abundance/biomass
     comparison (ABC)  method applied to macrozoobenthic communities
     living on  tidal  flats  in the Dutch  Wadden Sea.   Mar. Ecol.
     Prog. Ser.  In press.

Boesch, D.F.   1973.   Classification and community  structure  of
     macrobenthos of the Hampton  Roads area, Virginia.  Mar. Biol.
     21:226-244.
                               200

-------
Boesch,  D.F.  and  N.N.  Rabalais.  1987.  Long-term  environmental
     effects of offshore oil and gas development. Elsevier Applied
     Science, London and New York, 708pp.

Brillouin, L.   1962.   Science and information  theory.   Academic
     Press, New York, 169pp.

Cabioch,  L.,  J.C.  Dauvin,  and F.  Gentil.   1978.    Preliminary
     observations on pollution of the  sea  bed and disturbance of
     sublittoral communities in Northern Brittany by oil from the
     Amoco Cadiz. Mar. Pollut. Bull.  9:303-307.

Clifford,  H.T.,  and W.  Stephenson.    1975.   An  introduction to
     Numerical Classification. Academic  Press,  New York, 229pp.

Day, J.  H.,  J.G.  Field and  M.P.  Montgomery.   1971.  The  use of
     numerical methods to determine the distribution  of the benthic
     fauna across  the  continental shelf  off  North  Carolina.   J.
     Animal Ecol. 40:93-123.

Dunn,  O.J.    1964.    Multiple  comparisons  using  rank  sums.
     Technometrics 6:241-252.

Fager,  F.W.   1972.   Diversity:    a  sampling  study.   Am.   Nat.
     106:293-310.

Feder, H.M. and S.C. Jewett.   1981.   Feeding interactions in the
     eastern Bering  Sea with emphasis on the benthos.   In D. W.
     Hood and J. A.  Calder  (eds.), The Eastern Bering Sea Shelf:
     Oceanography and Resources.  U.S. Dept.  Commerce 2: 1229-1261.

Feder, H.M. and S.C.  Jewett.  1987.  The subtidal benthos.  In D.W.
     Hood and S.T.  Zimmerman (eds.),  The  Gulf  of Alaska.  Physical
     Environment and Biological Resources,  Ocean Assessment Div.,
     Alaska Office, U.S. Minerals Management Service,  Alaska OCS
     Region, MMS 86-0095, U.S. Govt.  Printing Office, Washington,
     D.C., pp 347-396.

Feder, H.M. and G.E.M. Matheke.   1980.  Distribution, abundance,
     community structure and trophic relationships of the benthic
     infauna of the northeastern Gulf of Alaska.  Inst. Mar. Sci.
     Report R78-8, Univ. Alaska,  Fairbanks, 211pp.

Feder,  H.M.,  G.J.  Mueller,  M.H.  Dick  and  D.B. Hawkins.   1973.
     Preliminary benthos survey, pp  305-386.   In  D.W. Hood, W.E.
     Shiels and E.J. Kelley  (eds.), Environmental Studies in Port
     Valdez.   Inst.  Mar.  Sci. Occas.  Pub.  No. 3,  Univ. Alaska,
     Fairbanks, Alaska 495 pp.

Field,  J.G.   1969.   The use  of  numerical methods  to determine
     benthic distribution patterns  from dredgings  in False Bay.
     Trans. Roy. Soc. S. Africa 39:183-200.

                               201

-------
Field, J.G.  1971.  A  numerical  analysis  of  changes in the soft-
     bottom fauna along a transect across  False Bay, South Africa.
     J. Exp. Mar. Biol. Ecol. 7:215-253.

Field, J.  G.,  and  G.  MacFarlane.    1968.   Numerical  methods in
     marine ecology.   I.  A  quantitative "similarity"  analysis of
     rocky shore samples in False Bay, South Africa. Zool. Africa
     3:119-253.

Gower, J.C.   1967.   Multivariate analysis  and  multidimensional
     geometry.   Statistician 17:13-28.

Gower, J.C.    1969.   A  survey  of   numerical  methods useful in
     taxonomy.   Acarologia 11:357-375.

Gray, J.S.  1989.   Effects of environmental stress in species rich
     assemblages.  Biol. J.  Linnean Soc. 37:19-32.

Gray, J.S. and  F.B. Mirza.  1979.  A  possible method for detecting
     pollution-induced disturbance on marine benthic communities.
     Mar. Pollut. Bull. 10:142-146.

Gray,  J.S.  and  T.H.   Pearson.    1982.    Objective selection of
     sensitive species  indicative of pollution-induced change in
     benthic communities. 1. Comparative  methodology.  Mar. Ecol.
     Prog. Ser. 9:111-119.

Gray, J.S., M.  Aschan,  Mr. R. Carr,  K.R. Clarke,  R.H.  Green, T.H.
     Pearson, R. Rosenberg,  and R.M.  Warwick.  1988.  Analysis of
     community   attributes   of   the  benthic   macrofauna   of
     Frierfjord/Langesundfjord and in a  mesocosm experiment.  Mar.
     Ecol. Prog. Ser.  46:151-165.

Heip, C.R., M.  Warwick, M.R.  Carr, P.M.J.  Herman, R. Huys, N. Smol
     and K. VanHolsbeke.  1988.  Analysis of community attributes
     of the benthic meiofauna of Frierfjord/Langesundfjord.  Mar.
     Ecol. Prog. Ser.  46:171-180.

Hoberg, M.K. 1986.  A numerical analysis of the benthic infauna of
     three  bays  in Prince William Sound, Alaska.  M.A.  Thesis,
     Humboldt State University, Arcata,  CA 153pp.

Hogan, M.E. and D.B. Irons.   1988.  Waterbirds and marine mammals.
     In D. G. Shaw and  M.  J.  Hameedi:  (eds.) Environmental Studies
     in Port Valdez, Alaska.  Springer-Verlag, Berlin: 225-242.

Hurlbert,  S.H.   1971.   The nonconcept of species diversity:   a
     critique and alternative parameters.  Ecology 52:577-586.

Kineman,  J.J.,  R.  Elmgren and  S.  Hansson.  1980.   The Tsesis Oil
     Spill.  U.S.  Dept. of  Commerce, Office  of  Marine Pollution
     Assessment, NOAA,  Boulder, CO,  296pp.

                               202

-------
Kruskal, W.H. W.A. Wallis.   1952.   Use of ranks in one criterion
     variance analysis.  J. Amer. Stat. Assoc. 47:583-621.

Lance,  G.N.,  W.T.  Williams.     1966.    Computer  programs  for
     hierarchical    polythetic    classification    ("similarity
     analyses").  Comput. J. 9:60-64.

Loya,  Y.    1972.   Community structure and species  diversity of
     hermatypic corals at Eilat,  Red Sea.  Mar. Biol. 13:100-123.

Margalef,  R.    1958.    Information  theory  in ecology.   General
     Systems 3:36-71.

McRoy, C.P.  1988.  Natural and anthropogenic disturbances at the
     ecosystem  level.  In  D.G.  Shaw  and  M.J.  Hameedi  (eds.).
     Environmental Studies  in  Port Valdez,  Alaska.   Springer-
     Verlag, Berlin:  329-344.

Mueller-Dombois, D. and H.  Ellenberg.  1974.  Aims and Methods of
     Vegetation Ecology Wiley,  New York, 547pp.

Nybakken, J.  1978.  Abundance, diversity  and temporal variability
     in a California intertidal nudibranch assemblage. Mar. Biol.
     45:129-146.

Odum, E.P.  1975.   Ecology.  Holt, Rinehart  and Winston, New York,
     244pp.

Pearson, T.H.  1975.   Benthic ecology of Loch Linnhe and Loch Eil,
     a sea loch system on the west coast of  Scotland.  IV. Changes
     in the benthic fauna  attributable to organic enrichment.  J.
     Exp. Mar.Biol. Ecol.  20:1-41.

Pearson, T.H. and R.  Rosenberg.   1978.  Macrobenthic succession in
     relation to  organic enrichment and  pollution  of  the marine
     environment.  Oceanogr. Mar. Biol. Ann. Rev. 16:229-311.

Peet, R.K.  1974.   The measurement of species diversity.  Ann. Rev.
     Ecol. Syst. 5:285-307.

Pielou, E.G.  1966a.   Species-diversity and pattern-diversity in
     the study of ecological succession.  J. Theor. Biol. 10:370-
     383.

Pielou, E.G.   1966b.   The  measurement of diversity in different
     types of biological collections.  J.  Theor. Biol. 13:131-144.

Pielou, E.G.  1977.  Mathematical Ecology.   Wiley, New York, 285pp.

Rosenbert, R.   1972.   Benthic  faunal recovery in a Swedish fjord
     following the closure  of a sulphite pulp mill.  Oikos 23:92-
     108.

                               203

-------
Sager, P.  and A.C.  Hasler.  1969.  Species diversity in lacustrine
     phytoplankton.  I. The components of  the  index of diversity
     from Shannon's formula.   Am. Nat. 102:243-282.

Sanders, H.L., J.F. Grassle,  G.R. Hampson,  L.S. Morse, S. Garner-
     Price, and C.C. Jones.  1980.  Anatomy  of an oil spill:  long-
     term effects from the grounding of the barge Florida off West
     Falmouth, Mass.   J. Mar.  Res. 38:265-380.

Shannon, C.E. and  W.  Weaver.   1963.   The  mathematical theory of
     communication.  Univ.  Illinois Press,  Urbana,  177pp.

Siegel, S.   1956.  Nonparametric statistics  for  the behavioral
     sciences.  McGraw-Hill,  London,  312pp.

Simpson,  E.H.    1949.    The  measurement  of diversity.    Nature
     163:688.

Sokal, R.R.  and F.J.  Rohlf.   1969. Biometry.   W.H.  Freeman,  San
     Francisco, California,  776pp.

Stephenson, W. and W.T.  Williams.  1971. A  study of the benthos of
     soft  bottoms.   Sek Harbour,  New Guinea, using  numerical
     analysis.  Aust.  J. Mar.  Freshwater Res.  22:11-34.

Stoker, S.  1978.  Benthic invertebrate macrofauna of the eastern
     continental   shelf  of   the  Bering/Chukchi   Seas.     Ph.D.
     Dissertation, Inst. Mar.  Sci., Univ. Alaska Fairbanks, 259pp.

Warwick, R.M.  1986.   A new method for detecting pollution effects
     on marine macrobenthic  communities.  Mar.  Biol. 92:557-562.

Warwick,  R.M.    1988.   Analysis  of  community  attributes  of  the
     macrobenthos of Frierfjord/Langesundfjord  at taxonomic levels
     higher than species.  Mar. Ecol. Prog. Ser. 46:167-170.

Warwick, R.M.,  T.H. Pearson and  Ruswahyuni.  1987.  Detection of
     pollution effects on marine  macrobenthos:  Further evaluation
     of the species abundance/biomass method.  Mar. Biol. 95:193-
     200.

Zar, J.H.  1974.  Biostatistical Analysis.  Prentice-Hall, Englewood
     Cliffs, New Jersey, 620pp.
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                              BUDGET

Salaries                    $467.7
Travel                        19.6
Contracts                     90.8
Supplies                      11.9
Equipment                      2.5
Total                       $592.5
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SUBTIDAL STUDY NUMBER 3

Study Title:  Bio-availability and Transport of Hydrocarbons

Lead Agencies:  NOAA, DEC


                           INTRODUCTION

This study  will continue  to  assess  the geographic  and temporal
distribution of dissolved and particulate hydrocarbons in the water
column resulting  from the EVOS.   Caged mussels will be  used to
determine the bio-availability of suspended hydrocarbons.  Sediment
traps provide a measure of suspended  load as  storms  and clean up
activities expose remaining shoreline oil deposits to weathering.

Analysis of caged mussels  at impacted  sites will compare levels of
petroleum hydrocarbons with levels in  mussels at unimpacted sites.
Levels of  hydrocarbons in mussel tissue  will demonstrate  that
hydrocarbons  are  biologically  available to  biota  in  nearshore
waters.

In 1991,  NOAA/NMFS will continue caged mussel deployments.  Field
efforts will be reduced by placing mussels at ten sites in PWS for
two one month exposures,  in addition  to collection of indigenous
mussels at transplant sites.

In 1991, NOAA/NMFS will also begin the  synthesis and interpretation
of hydrocarbon  contamination  data for mussels  and seawater from
seven NRDA projects.   This synthesis  will provide information on
hydrocarbon exposure  over  a broad geographical  area  and temporal
duration.

DEC conducted two retrieval cruises in 1990 for the original set of
five sediment traps.  Ten  additional traps deployed in August 1990
will be retrieved in March 1991  after  winter storms and before the
spring plankton bloom.  Work  in 1991 will concentrate all fifteen
traps at five sites to allow more  intensive monitoring.  The traps
will be  retrieved  again   in  June after the  plankton  bloom and
removed in September before winter storms.


                           OBJECTIVES

A.   Evaluate trends in ambient water quality using  bioaccumulators
     Mytilus  trossulus  as surrogates for  chemical measurements.
     Estimate  concentrations  of  petroleum derived  hydrocarbons
     accumulated  by mussels transplanted for  1  or 2 months along
     the oil spill trajectory  such that  the  estimate  is within 25%
     of the actual concentrations 95% of the time.
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B.   Synthesize  all water  and  mussel  hydrocarbon  data in  the
     Technical  Services 1  database to  provide a  comprehensive
     geographic  and  temporal  picture   of  trends  in  petroleum
     hydrocarbon concentrations in the near shore water column.

C.   Determine  if  sediments settling out of the water  column in
     nearshore subtidal environments contain adsorbed hydrocarbons.

D.   Decipher subtidal oiled sediment transport mechanisms through
     analysis of benthic sediments  and  stratigraphic analysis of
     bottom cores.
                             METHODS

NOAA/NMFS

Experimental Design

Prior to a new deployment cruise, bay mussels, Mytilus trossulus,
will be collected from a hydrocarbon free site on Admiralty Island
in southeast Alaska.  The mussels will be transported to Auke Bay
Lab and  held in living  stream  tanks that have been  rinsed with
dichloromethane and flushed with ambient unfiltered seawater at the
rate of 2 liters per minute  at least overnight.  Since mussel size
may influence hydrocarbon uptake (Bayne et al. 1981), only mussels
with shell length of 45-50 mm will be selected for deployment. At
least 30 animals from each collection will be sampled to determine
the population's base hydrocarbon level and condition.

Mussels will be kept aboard  the deployment  vessel in coolers and
the blue ice changed daily for up to 6  days.  A mussel "cage" is a
nylon mesh diver collecting bag.  For deployment,  20 mussels will
be placed on a rigid perforated polypropylene sheet fitted into the
bottom of each cage.  Assuming some mortality during exposure, this
number was selected to provide at least triplicate  samples of 10 g
of tissue for hydrocarbon  analysis.   On site,  a cage will be
attached to an anchored mooring  line at 1 m, 5 m, and 25 m depths.
The 2 shallower cage depths  were selected to correspond to water
column depths sampled by this study in  the first 6 weeks after the
spill; mussels  at  the third  depth  will be exposed to  the water
column about 10 m above  the  bottom at  low tide.   Mussels will be
exposed  for  approximately 30 days.   At  the conclusion  of each
deployment cruise another baseline mussel sample will be taken.

Details  of  deployment,  exposed  mussel  collection,  and  sample
handling are provided in Air/Water 3 Study Plans 1989 and 1990.

Sampling 1991

Mussels will be  deployed at ten 1990 sites within PWS.  Eight sites
were in the spill trajectory and subject to maximum original oiling

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as  indicated by  preliminary  analysis  of  water column  samples
(Air/Water 3), sediment pore water  samples  (Air/Water 4),  and by
DEC  Shoreline  Impact Composite  Maps.    All sites  coincide  with
Air/Water 2 sites and five coincide with Air/Water 3 sediment trap
locations.  There are two reference sites. Deployment  in 1991 will
indicate changes in hydrocarbon concentrations at these sites since
deployed mussels were last collected in September 1990.  Additional
mussels may  be  collected  in 1991 at specific sites in  PWS where
hydrocarbon data is needed.

Data Synthesis

The geographic  and temporal extent  of  water and mussel  samples
collected, and of those  submitted for hydrocarbon analysis will be
determined.  Samples that  have  not yet been selected for analysis,
but that may be needed to  provide a more complete documentation of
overall exposure  levels, will  be identified.   Additional  mussels
may be collected in 1991 at specific sites in PWS where hydrocarbon
data is needed.

Data Analysis

Analysis  of variance   (ANOVA)  will  be  used  to   determine  the
statistical  differences of hydrocarbons found  in  samples.  ANOVA
will also  be used to examine differences among water and mussel
samples in the data synthesis process.

Draft  graphic  presentations of  the data synthesis of all  NRDA
mussel samples will  be  prepared at Auke Bay  Lab with Munmap and
Autocad.  Final maps will  be prepared by Technical Services No. 3.

DEC

Experimental Design

The sediment trap design incorporates guidelines  developed from
previous  sediment trap  work  with  open-ocean  moored traps  and
laboratory flume studies (Woods Hole 1989).  The  original design of
the traps was only intended to capture sediments in the nearshore
subtidal  habitat  to   show presence   or  absence  of  adsorbed
hydrocarbons, without quantification of  flux rates.   This  is a
result of  presence of the  traps being deployed in the complex,
multidirectional, oscillatory current and wave environment of PWS
making control of variables difficult.  The sedimentary processes
occurring in the  area of  a trap may be difficult or exorbitantly
expensive to monitor.

Theoretically, estimation of trapping efficiencies in the field may
be determined by use of three parameters:

     (1)  Reynolds Number,  a  function  of current  speed  and the
          ratio of fluid viscosity to fluid density,

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      (2)  aspect ratio  (A) of height (H) to diameter (D), and
      (3)  the ratio of  flow speed to particle fall velocities.

In  short,  the  direction and  velocity of  any  currents  and the
geometry  of the  trap  (aspect ratio  and  axial  symmetry)  will
determine  if the  trap disrupts  the flow  field and  results in
turbulent eddies within and around the trap that will change any
naturally occurring  sedimentation patterns.  The  spacing of the
traps  determine  whether  they  affect  each  other's  trapping
efficiencies.

Based  on the  lack of  data regarding  currents,  the  traps  were
designed so that the aspect ratio, symmetry, and spacing would be
adequate for a variety  of conditions.  The trapping cylinders are
constructed  of Schedule  40,  high  chemical resistance  PVC,  (6"
inside diameter and 48" tall).   A  baffle of  0.5" square grid, 0.5"
deep  fits flush with the top of  each  trap.   These cylinders are
mounted on  a 20"  x 20" square  base, with rebar  extending 24" on
which the cylinders  are clamped.   Each trap suite contains three
cylinders.  Design considerations follow the  Woods Hole report
(1989) including:

      (1)  a cylindrical geometry for axial symmetry which promotes
          trapping efficiency;

      (2)  an 8:1  aspect ratio  to  minimize eddies,  reduce in-trap
          flow, and allow for a tranquil layer within the trap for
          current velocities to 20  cm/sec  (0.39  knots).   (In the
          sheltered bays  where  most traps are deployed, currents
          are probably  within this range);

      (3)  a base in a triangular configuration that is oriented to
          wave-induced shore-normal currents.  Cylinders are spaced
          at 18" centers and aligned to reduce chances any cylinder
          would be downstream of  another; and

      (4)  leveling  after deployment to maintain  orientation to
          currents and  the water  column.

Sampling 1991

The 1991 sampling plan will locate  traps  along  a transect to the
shore at three different depths.   Fifteen sediment traps will be
deployed at five sites in 1991.  At  each site, divers will place a
suite of traps at 10,  15 and 20 meters below MLLW.

The sediment traps are designed and located to collect sediments
settling from  the water column at  single points throughout PWS.
Coordination with other studies provides for result extrapolation
both spatially and temporally.  The trap  sites  have been matched
with sites used by Coastal Habitat previous DEC subtidal sampling,
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Subtidal 2, and NOAA caged mussels.  Sediment chemistry data will
thus be available over time and from a larger area.

Knowledge  is  derived of current  directions  and velocity  at the
sediment trap sites from qualitative observations of sedimentation
structures and drift patterns by  the  field team.   Particle size,
settling velocities,  and  current measurements will  aid  in the
differentiation of  bed-load movement  versus  resuspension  (Visher
1969; Middleton 1976),  delineation of erosional and depositional
events (Sundborg 1956), as well as allowing  calculations of trap
efficiency.   Differentiating  between  new  sediment input  to the
subtidal and cycling of  previously deposited sediments will give a
better understanding of localized  transport processes.  Due to the
great distances fine sediment particles can travel before settling
out of the water column (in a current flow of lOcm/sec,  a 0.06mm
silt particle may  travel  as  far as 10 km before  settling at 100
m.),  coordination  with Subtidal  No.  1 deepwater  sampling  is
emphasized.

Data Analysis

Particulate samples from the sediment traps  will  be screened for
hydrocarbon content by ultraviolet fluorescence spectrophotometry
after  methylene  chloride extraction of  samples.    UVF  is  a
semiquantitative method of analysis for hydrocarbons (ASTM 1982).
Samples showing significant quantities of  petroleum hydrocarbons
will be  further analyzed  for  polynuclear aromatic  hydrocarbons
(PAH)  and  total   petroleum  hydrocarbons   (TPH)   according  to
procedures established by Technical Services  Study No.  1.

Particle size analysis will be performed by sieving the sample in
a stacked set of Wentworth grade sieves to 62 urn.  Analysis of the
silt-clay fraction will  be obtained by pipette analysis.  Sediments
will  be  inspected for  composition,   and  cores for  sedimentary
structures.
                           BIBLIOGRAPHY

ASTM D-3650-78.  Standard test method for comparison of waterborne
     petroleum oils by fluorescence analysis.

Bassin,  N.J.,  and  T.  Ichiye,  1977.  Flocculation behaviour of
     sediment and oil emulsions. J. Sedim. Petrol. 47(2): 671-677

Bayne,  B.L.,  K.R.  Clarke and M.N.  Moore.   1981.  Some practical
     considerations  in  the measurement  of  pollution  effects on
     bivalve molluscs and some possible ecological consequences.
     Aquatic Toxicolgy 1:159-174.
                               210

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Blount, A., 1978.  Two years after the Metula oil spill, Strait of
     Magellan, Chile - oil interaction with coastal environments.
     Tech. Rept. No.  16-CRD,  Coastal Research Division, Dept. of
     Geology, Univ. of South Carolina, Columbia, S.C., 214 p.

Boehm, P.O., M.S. Steinhauer, D.R. Green, B. Fowler, B. Humphrey,
     D.L.  Fiest,  W.J.  Cretney.  1987.    Comparative  fate  of
     chemically  dispersed  and  beached  crude oil  in  subtidal
     sediments of the arctic nearshore.  Arctic 40 (1): 133-148.

Conover, R. J.  1971.  Some relations between zooplankton and Bunker
     C  oil in Chedabucto Bay  following the wreck of  the tanker
     Arrow. J. Fish. Res. Bd., Can. 28: 1327-1330.

Gundlach, E.R., C.H. Ruby, L.G. Ward, A.E. Blount,  I.A.  Fischer and
     R.J.  Stein. 1978.   Some guidelines for oil-spill control in
     coastal  environments  (based  on  field studies of four oil-
     spills) In Proc.  of  1977 ASTM sympos. on chem. dispersants for
     the control of oil spills.   Amer. Soc. Testing and Materials,
     Philadelphia, Penn.  32p.

Gundlach,  E.R.,  P.D.  Boehm, M. Marchand,  R.M.  Atlas,  D.M. Ward,
     D.A. Wolfe. 1983.  The fate of Amoco Cadiz oil.  Science 221:
     122-129.

Middleton,  G.V.  1976.   Hydraulic  interpretation  of  sand size
     distribution.  J. Geol. 84: 405-26.
Sundborg,  A.  1956.    The river  Klaralven:
     processes.  Geogr. Ann. 38: 127-316.
                          a  study of  fluvial
Visher,  G.S.  1969.   Grain  size distributions  and depositional
     processes.   J. Sed. Petrol. 39: 1074-1106.

Woods  Hole  Oceanographic   Institution  1989.     Sediment  trap
     technology  and sampling.   U.S.  Global  Ocean  Flux Planning
     Report Number  10, August, 1989.
Salaries
Travel
Contractual
Supplies
Vessel

TOTAL
NOAA

110.0
 21.0
  0.0
 19.0
  0.0
 BUDGET

  DEC

$69.0
 10.3
 11.5
  2.2
103.2
150.0   $196.2
 Totals

$179.0
  31.3
  11.5
  21.2
 103.2

$346.2
                               211

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SUBTIDAL STUDY NUMBER 4

Study Title:   Fate and  Toxicity of  Spilled  Oil From  the Exxon
               Valdez

Lead Agency:   NOAA


                           INTRODUCTION

Overview and Relation to other Studies

This study  is  designed:   a)  to assess the  toxicity  of weathered
Exxon Valdez  oil and its  degradation products to selected test
organisms;  and   b)  to integrate the  results  from  selected other
projects, both within  and outside the NRDA,   into an overall budget
for the distribution, transport, transformation, and persistence of
spilled oil in Alaskan coastal  environments.   The study is very
closely coordinated with Subtidal Study No. 1 for  its field work
and toxicity studies,  and will require close interaction with all
of the present  and  past Air/Water studies,   the Coastal Habitat
studies,  and with related spill  response studies for completion of
the spilled oil budget.

Toxicity of Prudhoe Bay Crude Oil and its Products  of Weathering

Very limited information  is available on the significance of either
the polar constituents of crude oil or the intermediate oxidation
products of petroleum  hydrocarbons  (whether  from photooxidation or
biodegradation) in terms  of their potential  for bioaccumulation and
toxicity to resource  organisms  in the marine  environment.   Since
these  compounds  have  undergone   preliminary  oxidation  and
(sometimes) conjugation,  they are more  polar than  their  parent
hydrocarbons, and will  as  a result generally be more subject to
excretion  or  depuration,  less  subject  to  bioaccumulation,  more
susceptible   to   further    oxidation   (or  biodegradation   if
accumulated), and more susceptible to dilution  and dispersion in
the water  column.   A  detailed review of the  literature on these
topics was included  as part of the study plan for this project last
year.   Under this  project very  limited studies were initiated
during 1990 to  determine whether such polar  constituents  pose a
significant risk of toxicity  or mutagenicity to  Alaskan  marine
organisms as a result of the EVOS.

Acute Toxicity of Ambient Spilled Oil to Marine Organisms

Last year's study plan provided a review of the very considerable
body of literature that exists on the toxicity  of Alaskan crude oil
to  Arctic  and  subarctic  marine organisms.   The  data base is
probably  adequate for  assessing the relative  sensitivities of
different marine species to exposure and for estimating the range
of   potential  responses (at the organism  level) that may result

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from  a particular level  of exposure  in  the environment.   Very
little of this prior research on toxicity was directed, however, at
the  specific contribution  of  either hydrocarbon  metabolites or
other  oxidation products  of  oil  that may be  produced  by  the
processes of biological or chemical weathering in the environment.

Much of the early work  in this area focused on the acute toxicities
(generally 96-hour exposures) of water-soluble fractions (WSF) of
fresh Cook Inlet crude oil and Prudhoe Bay crude oil to a variety
of  species  and life  stages of  commercially or  recreationally
important Alaskan marine organisms.  Data on the acute toxicities
of crude oil to marine organisms of interest have been summarized
by Brodersen et al. (1977), Craddock (1977), Moles et al.  (1979),
Rice  et al.  (1976,  1977,  1979, 1984),  and National  Academy of
Sciences   (1985).      Rice  et al.  (1981)  demonstrated  that  the
compositions  of the water-soluble fractions of  Cook  Inlet  and
Prudhoe Bay crude oils  were very similar both to one another and to
that  of the  discharge from  the  ballast  treatment  facility at
Valdez.

Sublethal  effects  of oil   exposure   have  also  been  studied
extensively, through the use of long-term exposures (e.g., up to 40
days) to  WSF of Alaskan  crude  oil,  or of  prolonged  exposure to
oiled  food or  oiled sediments.     Earlier work   (which focused
primarily on temperate organisms and crude  oils from sources other
than Alaska) was summarized  by  Anderson (1977),   Johnson  (1977),
and   Patten  (1977).   During the  late 1970's and  early 1980's,
increased  attention was given  to arctic and subarctic organisms,
especially relative to Alaskan and  Canadian oils, and some of this
more recent  work has  been reviewed by Rice  et al.  (1984) ,  Rice
(1985), Wolfe  (1985),   National  Academy of Sciences (1985),   and
Karinen (1988).

In conjunction with Subtidal Study No. 1, work was undertaken under
this  project  in  1990  to  test  the ambient  toxicity  of  marine
sediments  from  PWS and the  nearby  GOA  to  two bioassay organisms:
the marine amphipod  Ampelisca abdita  and  the  oyster Crassostrea
gigas.   Although  results of  this work have  not  been analyzed
completely, preliminary results indicate that sediments from oiled
sites in PWS were significantly more toxic  to the bioassay species
than were sediments from unoiled or lightly oiled reference  sites.

Fate of Spilled Oil: Budgets and "Mass Balance"

An accurate and complete mass balance is difficult to assemble for
a  major oilspill  in  the  marine  environment.    The  quality of
estimates  of the  quantities and  locations of  oil  affected by
different processes of  transport or transformation have varied from
spill to spill, depending on the local circumstances of the spill
and  the  level of  effort  devoted  to any particular  process.
Selected observations at past spills have been summarized by Mackay
(1981), Gundlach et al. (1983), Jordan and Payne (1980), National

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Academy of Sciences (1985),  and Wolfe  (1985, 1987) .    Information
especially pertinent for summarizing the fate of oil from the EVOS
has been and is  still  being gathered  by the Interagency Response
Team and the DEC, and by certain projects  under the NRDA Program:
especially  Coastal  Habitats  Studies  1&2,   A/W  Studies  1-5,
Fish/Shellfish  Study 24  and Technical Services Study  1.   Oil
weathering models  (Payne 1983,  1984)  and  transport/fate  models
(Gait and Torgrimson 1979; Spaulding et al. 1983), constructed to
predict the  distribution and  fate of spilled oil,  should also
provide valuable insight and assistance in preparation of a budget
for the oil spilled by the Exxon Valdez.


                            OBJECTIVES

A.   Document the toxicity  of  contaminated sediments  and related
     environmental samples to selected marine biota

B.   At selected sites,  document and quantify the  occurrence of
     oxidized derivatives of Exxon  Valdez  oil;  and determine the
     extent to  which the  observed toxicity of  oil-contaminated
     environmental  samples  may  be  attributable  to  oxidation
     products of petroleum.

C.   Construct a summary budget or "mass balance" summarizing the
     fate of the spilled oil.
                             METHODS

A.   Toxicity of Oil-Contaminated Sediments And Other Environmental
     Samples

A boat-based survey of surficial sediment toxicity was carried out
in 1989 under A/W Study No. 4,  at all stations sampled during June
to August, 1989 (Leg II) .  The toxicity bioassay  used in that study
was  the standard  Microtox assay,   in  which  a  composite  of  the
replicate  sediment  samples obtained  at  each   depth  from  each
sampling  site is  analyzed for  sediment  toxicity  based  on  the
inhibition of bioluminescence in Photobacterium phosphoreum  (15-min
Microtox assay).  Organic extracts of the sediments were prepared
and assayed for toxicity by the methods of Schiewe et al (1985).
The Microtox  assay is rapid,  simple,  inexpensive,  and sensitive;
and the bioassay results have correlated well in  other studies with
the results of other  standard bioassays  that use fish, amphipods
or bivalve larvae as  test organisms (Chang 1981,  Williams et al.
1986,  Giesy  et  al.  1988).   Results  of  the  1989  survey  also
correlated with UV fluorescence analyses  of  oil in  the sediment
samples.

Under A/W 6,  toxicity  tests were  performed  in  1990  on sediment
samples taken at selected  sites sampled by A/W Study No.  2 from the

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NOAA  ship  Davidson.      Two  specific  tests,  both  following
well-established protocols, were used:  a sediment elutriate test
using larval  oysters,  and a whole sediment  test using Ampelisca
abdita.     Crassostrea is  a standard  bioassay species  used  to
represent  intertidal and  subtidal  bivalve species  whose larval
recruitment is  vulnerable to interruption by  toxic  oil residues
remaining  in  intertidal  sediments.    Ampelisca  inhabits  soft
nearshore  sediments  that  are  possible  sinks  for  petroleum.
Subtidal ampeliscid amphipods exhibited considerable sensitivity to
oil  in  the aftermath  of the Amoco Cadiz spill  (Cabioch  et al.
1982).  Use of these two species was intended to provide a direct
measure  of the  toxicity of  the  residual oil to actual marine
species.  Preliminary test results from the 1990 samples indicated
that sediments from oiled sites were  more toxic to both bioassay
organisms  than  were sediments from unoiled  reference  sites, and
both bioassays  are proposed to be repeated  in 1991  to determine
whether  the toxicity has persisted and how its  levels  may have
changed.

Detailed methods for both of the proposed tests  have been described
previously:   for the oyster larvae bioassay  (Chapman  and Morgan
1983; Chapman and Becker 1986);  and  for the Ampelisca test (Long,
Buchman et al. 1989; Scott and Redmond 1990).

Sediment  samples will  be collected  during  one  or  more  of the
sampling cruises described  under Subtidal Study No.  1.  Sampling
sites have been  selected to represent the more  heavily oiled areas
and a set  of  unoiled (or very lightly oiled)  reference sites for
comparison.  At each of 15 of the sites,  eight one-liter samples
of surficial sediments (top 5 cm) will be  collected (2 each at the
intertidal, 6-meter, 20-meter, and 100-meter depths)  for toxicity
testing  with  Crassostrea and Ampelisca.   These  samples will  be
stored  at  0-4°  C,  and  offloaded  from  the  vessel  at  regular
intervals  for shipment  to  a testing  laboratory to be selected
through  a  competitive contracting  procedure.   Bioassays will  be
initiated within 10 days of the collection of the samples.

B.  Oxidation Products of Petroleum

Two contracts were  initiated under this study  (A/W 6)  in 1990 to
determine the  presence and significance of polar oxidation products
of petroleum in the marine environment of PWS.

At two heavily oiled sites and one lightly or unoiled site in PWS,
special  samples were  taken  by  a team  of researchers from Science
Applications    International    Corporation,     to    assess   the
concentrations and  compositions of  petroleum oxidation products,
and their toxicity,  in intertidal sediments and  interstitial water.
Large quantities of sediments and interstitial  water were required
to support  the  necessary development  of  suitable techniques for
bulk fractionation of samples for chemical  characterization and
quantification of the polar metabolites and for toxicity testing.

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The intertidal sediments and interstitial water were extracted with
methylene chloride,  and  the extracts were subjected  to toxicity
testing with a suite of bioassays, including the standard Microtox
bioassay (Schiewe et al. 1985), the Ames mutagenicity test  (Ames et
al. 1975), the  SOS  Chromotest  assay for genotoxicity (Quillardet
and Hofnung 1985, Quillardet et al.  1985), and the Mytilus larval
toxicity bioassay.  The extracts were then fractionated to separate
polar from non-polar constituents,  and  the toxicity  of  the polar
fractions will  be compared with  the better known toxicities of
aromatic fractions and reference compounds.   Those fractions that
demonstrate  significant  toxicity will  be analyzed  by GC-MS to
identify the composition of polar constituents.

A second contract was  let to Bermuda Biological  Station  for the
analysis of selected mussel (Mytilus trossulus) tissue samples for
polar oxidation products  to ascertain whether these compounds were
present in, and bioaccumulated from, the oiled PWS environment.

At the time of this plan,  results were not available from either of
these contracts.  During 1991, the  contractors'  reports  will be
received and  evaluated, and final recommendations will be developed
on  how to assess  the probable toxicity  of polar  constituents
arising  from  the   EVOS.    These   studies   may   lead   also  to
recommendations for  analyses of  polar  constituents  to supplement
the   traditional   hydrocarbon   analyses  being   performed   on
environmental samples  taken by  other projects within the overall
NRDA.

C.  Budget for Fate of Spilled Oil

This task is primarily a  synthesis  function.   Information on the
distribution and  fates of  Exxon  Valdez  oil needs  to  be  assembled
from a  number of sources,  interpreted  in the light  of existing
information and models, and presented in a way that will support a
region-wide assessment of the potential effects of the spill.

During  1990,  a  small  Steering  Group  of spill  experts met to
identify the compartments and processes  that should be included in
the FATES budget.   The Steering Group  identified the  following
compartments for initial  analysis and inclusion in the budget:  1.
Water Surface (floating oil),   2. Intertidal Zone (stranded oil),
3.  Water  Column (dissolved and accommodated oil),   4. Subtidal
Sediments (sunken and settled oil, or oil otherwise transported to
bottom sediments),   5. Atmosphere  (evaporated oil).   The actual
masses of oil in these  different  compartments are quite different,
and because  of  transfers among  compartments as the  spill was
transported through  and out of  PWS, the pertinent time  and space
scales  are also  quite different.  As   a result,  very  different
estimation methods  have  been  used  (by  different  people)  for the
various compartments.  The Steering  Group concluded therefore  that
information for these  five compartments would best be synthesized
separately, with appropriate effort  to reconcile both the separate

                               216

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compartmental estimates as well as any  estimates of fluxes between
compartments  .    For  each compartment  and its  associated major
fluxes,  the Steering  Group  identified  and  discussed  important
sources of  data,  historical  information,  and modeling expertise,
and suggested preliminary courses of action, as summarized in the
report of the steering group meeting.

Potential sources of  data, historical  information,  and modeling
expertise were identified for:

1.   Floating oil  (distribution in Time & Space)
2.   Evaporation and atmospheric dispersion
3.   Photooxidation in the atmosphere
4.   Mousse formation
5.   Beaching of oil & mousse (T&S)
6.   Water column accommodation (T&S)
7.   Photooxidation in water column, in
     slicks and on beaches
8.   Biodegradation in water column
9.   Transport to subtidal sediments
10.  Biodegradation in sediments

Representatives of  the above noted  activities,  along with other
recognized experts on oil weathering and fates, will be consulted
for recommendations on  appropriate approaches to synthesis,  and for
their  judgments  on the  suitability  and  adequacy   of  existing
information for development of the FATES model.  Timely progress on
the  FATES budget  will depend  on the  availability  of  suitable
information from other sources and projects.   Chemical data, i.e.,
from TS No. 1, will be of utmost  importance to the completion of
this project.  Where existing information is found to be deficient,
means will be explored for gathering  of improved information.  The
reliability of all estimates will  be  assessed  and qualified in the
final analysis.

To the extent practical,  lead individuals will be designated for
coordination and completion of the synthesis related to each of the
identified compartments,  especially  where those compartments and
processes are  included  explicitly  in  the NRDA.    For  example,
initial assessment  of the hydrocarbon  levels  and weathering in
intertidal and subtidal sediments  will be  conducted under Subtidal
No. 1  (A/W  2), while the  assessment  of  water column  data will be
done under  Subtidal No.  3  (A/W  3) .   Effort should be  made to
identify all sources of relevant data and information for each of
the individual  compartments in the Fates budget.  The synthesis for
each of the compartments should include estimates of the rates of
transport  and  transformation processes   ongoing within  and/or
between compartments, including such processes as mousse formation,
Photooxidation,    biodegradation,    evaporation,    dissolution,
accommodation,   chemical  weathering  and  compositional  change,
"bleeding" of sheen, adsorption-sedimentation,  sinking, down-slope
transport  of  oiled  sediments,   etc.     Following  this  initial

                               217

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synthesis at the compartmenta1 level, the results will be brought
together at a summary  review and workshop to examine, explain, and
eliminate inconsistencies  among  data sets; and  to  encourage and
promote development of a single,  complete, and accurate consensus
synthesis product for all components together.  Every effort should
be made  in  advance of  the workshop to compare  and  reconcile the
independent  estimates of  inter-compartmental  transfers,  however,
these will be scrutinized  in  detail  at the workshop itself.   The
final synthesis will include detailed assessments of the quality of
the data and information,  including analytical confidence limits,
sampling adequacy  in time and space, and  model  reliability.   As
part of  this analysis,  the reliability of all  estimates  will be
assessed and qualified.    Efforts  will  also  be made to  present
estimates of oil distribution  in a form amenable to comparison with
existing  information  on  toxicity  to  facilitate any  subsequent
assessments of the potential effects on biological resources.

D.  Quality Assurance and Control

All   samples   will  be   taken   with    careful  adherence   to
Chain-of-Custody requirements.  All of the intertidal and subtidal
sediment samples analyzed  under this  study will be retained in the
custody of the laboratories performing the analyses, as called for
in  the  guidelines provided  by  the Technical Services  No.  1
Analytical Committee.  The detailed  protocols for   collection of
intertidal  and  subtidal   sediment  samples   are given  in  past
proposals for Air/Water Study No. 2.


                           BIBLIOGRAPHY

Ames,  B.N.,   J.McCann,  and  E.  Yamasaki.    1975.    Methods  for
     detecting carcinogens and mutagens with the  Salmonella
     mammalian microsome mutagenicity test. Mutation Research 31:
     347-364.

Anderson, J.W.  1977.  Responses to sublethal levels of petroleum
     hydrocarbons:   Are  they sensitive  indicators  and  do  they
     correlate with tissue contaminants?  PP.  95-114.   In:  D.A.
     Wolfe  (ed.)   Fate  and effects  of petroleum hydrocarbons in
     marine organisms and ecosystems.  Pergamon Press,  New York.

Brodersen, C.C., S.D. Rice, J.W. Short, T.A. Mecklenburg, and J.F.
     Karinen.  Sensitivity of larval and adult Alaskan shrimp and
     crabs to acute exposures  of the water-soluble fraction of Cook
     Inlet crude  oil.   PP  575-578.   In:   Proceedings,  1977 oil
     spill  conference  (Prevention,  Behavior,  Control,  Cleanup),
     American Petroleum Institute Publication No. 4284, Washington,
     D.C.
                               218

-------
Cabioch,  L.,  J.-C.  Dauvin,  C.  Retiere,  V.  Rivain,  and  D.
     Archambault. 1982.  Les  effets  des hydrocarbures de 1'Amoco
     Cadiz sur les peuplements benthiques des Bales de Morlaix et
     de  Lannion  d'Avril  1978 a  Mars  1981.   Pp.  205-228.    In
     Ecological study  of the Amoco Cadiz oil  spill.   Rpt of the
     NOAA-CNEXO joint scientific commission.  U.S. Dept. Commerce,
     NOAA.  Washington, D.C.

Chang,  J.C.,  P.B.  Taylor,  and F.R.  Leach.    1981.   Use  of the
     Microtox  assay  system  for  environmental  samples.    Bull.
     Environ. Contam. and Tox.  26: 150-155.

Chapman,  P.M.,  and S. Becker.  1986.    Recommended  protocols for
     conducting laboratory bioassays on Puget Sound sediments.  In;
     Final  Report    TC-3991-04.    U.S.   Environmental Protection
     Agency Region 10.  Seattle,  Washington.  55 pp.

Chapman, P.M., and   J.D. Morgan.   1983.  Sediment bioassays with
     oyster larvae.  Bull.  Environ. Contam. and Tox. 31:438-444.

Craddock, D.R.  1977.  Acute toxic effects of petroleum on arctic
     and  subarctic marine  organisms.   Pp 1-93.   In  D.C. Malins
     (ed.)   Effects  of petroleum on  arctic  and  subarctic marine
     environments and  organisms.   Vol.   II,   Biological Effects.
     Academic Press, New York.

Gait, J.A. and G M.  Torgrimson.  1979.  An on-scene spill model for
     pollutant trajectory simulations,  pp.  343-366.  In Proceedings
     of  the  workshop on physical  behavior of oil  in the marine
     environment. May  7-9,  1979, Princeton, NJ.,  Dept.  of Civil
     Engineering, Princeton Univ.

Giesy, J.P., R.L. Graney, J.L. Newsted,  C.J. Rosiu,  A. Benda, R.G.
     Kreis,  Jr.,  and  F.J.  Horvath.   1988.   Comparison  of three
     sediment  bioassay methods  using  Detroit  River sediments.
     Environ. Toxicol. & Chem. 7: 483-498.

Gundlach, E.R., P.O.  Boehm, M. Marchand,  R.M. Atlas,  D.M. Ward, and
     D.A. Wolfe.   1983.  The fate of Amoco Cadiz oil.  Science 221:
     122-129.

Johnson,  F.G.   1977.  Sublethal biological  effects of petroleum
     hydrocarbon exposures:   bacteria,  algae,  and  invertebrates.
     Pp  271-318.   In D.C.  Malins  (ed.)   Effects of petroleum on
     arctic and subarctic marine rnvironments and organisms. Vol.
     II, Biological Effects.  Academic Press, NY.

Jordan,  R.R.,  and  J.R.  Payne.  1980.   Fate  and  weathering  of
     petroleum  spills in  the marine environment:  A literature
     review  and  synopsis.    Ann Arbor  Science  Publishers.   Ann
     Arbor, Michigan.  174 pp.
                               219

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Karinen, J.F.  1988.  Sublethal effects of petroleum on biota.  Pp.
     294-328.  In D.G.  Shaw and M.J. Hameedi  (eds.)  Environmental
     studies  in  Port Valdez, Alaska:    A basis  for  management.
     Lecture  notes  on  coastal  and estuarine  studies,  Vol.  24.
     Springer-Verlag,  Berlin.

Lech,  J.J.  and  J.R.   Bend.  1980.    The  relationship  between
     biotransformation  and the  toxicity  and fate  of  xenobiotic
     chemicals in fish.  Environ.  Health Perspectives 35:115.

Long, E.R., M.F.  Buchman et al.  1989.  An evaluation of candidate
     measures  of  biological effects for the National  Status and
     Trends  Program.   NOAA Tech  Memo.  NOS  OMA  45.  NOAA Ocean
     Assessments Division,  Seattle,  WA.  106 pp. + appendices.

Mackay, D.  1981.   Fate and behaviour of  oil  spills. Pp. 7-27.  In
     J.B. Sprague, J.H.  Vandermeulen,  and P.G. Wells  (eds.)  Oil
     dispersants   in   Canadian    seas-research   appraisal   and
     recommendations.  Environment Canada,  Toronto.

Moles, A., S.D. Rice, and S. Korn.  1979.  Sensitivity of Alaskan
     freshwater and anadromous fishes to  Prudhoe Bay crude oil and
     benzene.  Trans. Am. Fish.  Soc. 108(4):  408-414.

National Academy  of  Sciences.  1985.   Oil in the  sea.   Inputs,
     fates, and effects.  National Academy Press,  Washington, D.C.

Patten,  B.C.   1977.    Sublethal  biological  effects of  petroleum
     hydrocarbon exposures:   fish.  Pp.  319-335.   In D.C. Malins
      (ed.)   Effects  of petroleum on arctic  and  subarctic marine
     environments and  organisms.   Vol.  II,   Biological Effects.
     Academic Press,  New York.

Payne, J.R.  1983.  Oil-weathering computer program for multivariate
     analysis of petroleum weathering in the marine environment-sub
     arctic.    Report  submitted   to  NOAA/OCSEAP,  Contract  No.
     NA80RAC00018.   Science Applications  Inc., La Jolla,  CA.  83
     pp.

Payne, J.R.  1984.   Multivariate  analysis of petroleum weathering
     in  the  marine  environment-sub arctic.    Vol. I  Technical
     Results.  Vol.  II.  Appendices.  Final Report submitted to
     NOAA/OCSEAP, Contract No. NA80RAC00018.  Science Applications
     Inc., La Jolla,  CA.  Multiple pagination.

Quillardet,  P.,  and  M.  Hofnung.  1985.    The SOS-Chromotest,   a
     colorimetric  bacterial  assay for  genotoxins:  Procedures.
     Mutation Research 147: 65-78.
                               220

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Quillardet, P., 0. Huisman, R.  D'Ari,  and M.  Hofnung.  1985.  The
     SOS-Chromotest, a colorimetric bacterial assay for genotoxins:
     Validation study with 83  compounds.   Mutation Research 147:
     79-95.

Rice, S.D.  1985.  Effects of oil on fish.  Pp. 157-182.  In F.R.
     Engelhardt   (ed.).     Petroleum   effects   in  the  arctic
     environment.  Elsevier Applied  Science Publishers,  New York.

Rice, S.D., J.W. Short,  and J.F.  Karinen.   1976.   Toxicity of Cook
     Inlet crude oil and No.  2 fuel oil to several Alaskan marine
     fishes and invertebrates.  Pp.  394-406.  In  Sources, effects
     &  sinks  of hydrocarbons  in the  aquatic environment.   The
     American Inst. of Biological Sciences, Washington, D.C.

Rice, S.D.,  J.W.  Short, and  J.F.  Karinen.   1977.   A  review of
     comparative oil toxicity and comparative animal sensitivity.
     Pp. 78-94.  In D.A. Wolfe  (ed.)   Fate and  effects of petroleum
     hydrocarbons  in  marine  organisms and  ecosystems.   Pergamon
     Press,  New York.

Rice, S.D.,  A.  Moles,  T.L.  Taylor,  and J.F.  Karinen.  1979.
     Sensitivity of 39 Alaskan marine species to  Cook Inlet crude
     oil and No. 2 fuel oil.   Pp.  549-443.  In Proceedings, 1979
     oil spill  conference (prevention, behavior, control, cleanup).
     American Petroleum Institute Publication No. 4308, Washington,
     D.C.

Rice, S.D.,   S. Korn,  C.C.  Brodersen,   S.A. Lindsay,  and S.A.
     Andrews. 1981.  Toxicity of ballast-water treatment effluent
     to marine organisms at Port Valdez,  Alaska.  Pp.  55-61.  In
     Proceedings, 1981 oil  spill conference (prevention, behavior,
     control, cleanup).   American Petroleum Institute Publication
     No. 4334,   Washington, D.C.

Rice, S.D.,  A.  Moles,  J.F.  Karinen,  S.  Korn, M.G.  Karls, C.C.
     Brodersen, J.A.  Gharrett,  and M.M. Babcock.  1984.  Effects of
     petroleum  hydrocarbons  on  Alaskan  aguatic  organisms:  A
     comprehensive review of  all oil-effects  research on Alaskan
     fish and invertebrates conducted by the Auke Bay Laboratory,
     1970-1981.    U.S.   Dept.  Commerce,     NOAA  Tech.   Memo.
     NMFS/NWC-67.  128 pp.

Scott,  J.K.,  and  M.S.  Redmond.     1990.   The effects  of  a
     contaminated dredged material on laboratory populations of the
     tubiculous amphipod Ampelisca  abdita.   In  U.M.  Cowgill and
     L.R. Williams (eds.) Aquatic toxicology and hazard assessment,
     Vol.  12.    ASTM  STP 1027.    Am.  Soc.  Testing  Materials,
     Philadelphia,  PA.
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Shiewe, M.H.,  E.G.  Hawk, D.I. Actor, and M.M. Krahn.   1985.  Use of
     a  bacterial  bioluminescence  assay  to  assess  toxicity  of
     contaminated marine  sediments.     Can.  J.  Fish,  and Aquat.
     Sci.  42:  1244-1248.

Spaulding, M.L., S.B.  Saila,  E.  Lorda, H. Walker, E. Anderson, and
     J.C.  Swanson.   1983.   Oil-spill fishery  impact  assessment
     model: Application  to  selected  Georges Bank fish species.
     Estuar. Coastal Shelf Sci.  16:511-541.

Williams, L.G., P.M. Chapman and T.C. Ginn.  1986.   A comparative
     evaluation  of  marine  sediment  toxicity  using  bacterial
     luminescence,  oyster embryo and amphipod sediment bioassays.
     Mar. Environ.  Res. 19:225-249.

Wolfe,  D.A.    1985.   Fossil  fuels:  Transportation  and  marine
     pollution.  Chapter 2.   Pp. 45-93.  In Iver W.Duedall, Dana R.
     Kester, P. Kilho Park and Bostwick H. Ketchum (eds.), Wastes
     in the ocean,  volume 4.  Energy  wastes in the  ocean.   John
     Wiley & Sons,  New York.

Wolfe,  Douglas A.    1987.     Interactions  of  spilled oil  with
     suspended  materials  and sediments  in aquatic systems.   Pp.
     299-316.  In K.L.  Dickson, A.W. Maki, and W.A. Brungs (eds.),
     Fate  and  effects of  sediment-bound  chemicals  in  aquatic
     systems.    Proc.  of  the 6th   Pellston  Workshop,  Aug 12-17,
     1984.  Florissant, Colorado.  Pergamon Press, Oxford, England.


                              BUDGET

Salaries                   $  24.0
Travel/shipping               11.0
Contracts                     85.0
Supplies                       5.0

Total                        $125.0
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SUBTIDAL STUDY NUMBER 5

Study Title:   Injury to PWS Spot Shrimp

Lead Agency:   ADF&G


                           INTRODUCTION

This project will  continue to determine possible  damage  to spot
shrimp, Pandalus platyceros, due to the  EVOS.   Spot shrimp are a
representative  species   of   the  deepwater   nearshore  benthic
ecosystem, serving as a food source for  a  variety  of fish.  They
are a commercially important species and also support subsistence
and personal use fisheries in PWS.  This project is a continuation
of F/S Study No. 15 which was conducted during 1990-91.

Spot shrimp are known to be sensitive to oil contamination in both
the larval and adult phase, and the effects of oil on spot shrimp
in particular and shrimp in general are well documented (Anderson
et al. 1981; Brodersen et al.  1977;  Brodersen 1987; Mecklenburg,
Rice and  Karinen  1977; Sanborn  and  Malins 1980;  Stickle  et al.
1987;  Vanderhorst   1976).     To  determine   the  . impacts  that
hydrocarbons from the spill may  have had on spot shrimp,  samples
will again be collected from the three  oiled and three non-oiled
sites  in western  PWS  which  had  been  surveyed  in  1990.    An
additional site will be used in  1991 to  increase the sample size
for  fecundity  and  modal  analysis  in the  oiled area.  The data
collected from  the  samples will be analyzed  to determine tissue
hydrocarbon levels  and tissue damage.  The collected data will also
be tested to  confirm or  reject  the  hypothesis that  there is no
significant difference in  hydrocarbon levels between the oiled and
non-oiled areas.  Relative abundance, in terms of  catch per unit
effort, at each study site and changes in relative abundance over
time will be tested  to determine possible relationships with the
level of oiling. A comparison with historical records will also be
made.   The  size composition  of  the stock  at each  site  will be
estimated and,  dependent  upon recruitment  to the  fishing gear,
analyzed to determine whether the 1989 year class suffered a high
mortality rate in areas of high oil impact relative to other year
classes in non-oiled areas.   Spot shrimp fecundity will  also be
determined  and tested  for  significant  interannual  differences
between oiled and non-oiled sites.
                            OBJECTIVES

     Estimate the  relative abundance by  weight and  sex  of spot
     shrimp and the  relative  abundance  by weight of  incidentally
     caught pink and coonstripe shrimp in oiled and unoiled areas
     and compare these values to those obtained during surveys in
     1989 and 1990.

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B.   Compare size and age frequencies of  spot  shrimp (by sex and
     depth stratum)  between sites using mixture modal analysis.

C.   Estimate  fecundity,  egg  mortality,  and  other  sublethal
     effects  between oiled  and  unoiled  areas over  time,  and
     determine whether those effects result in adverse changes in
     reproductive viability.

D.   Analyze tissue and  egg  samples  for presence of hydrocarbons
     and compare differences  between  oiled and unoiled sites. Test
     the hypothesis that the level of hydrocarbons is not related
     to the level of oil contamination present at a site.

E.   Document injury  to  tissues and compare differences between
     oiled and unoiled sites if warranted by results from tissue
     hydrocarbon analysis.

F.   Provide information  on stock status, hydrocarbon concentration
     and other  indicators  of stock condition  for  restoration of
     damages  and management of  the  spot shrimp  resource  for
     subsistence, personal and commercial user groups.


                             METHODS

This project uses commercial spot shrimp pots  of  a standardized
size to  catch spot  shrimp  in oiled and  unoiled areas.   Shrimp
specimens will be analyzed  for Prudhoe Bay crude  oil  levels and
necropsied to determine  if damage has  occurred to tissues  as a
result of oil contamination.   Only one sampling period will occur
during the winter of 1991-92.  The sampling period will take place
in early November (1991)  following the fall molt and egg extrusion.
Relative abundance estimates of spot shrimp will be made using a
stratified pot  deployment based on  depth and  location.   Size
distribution, species composition, and reproductive data will also
be collected.  Previous spot shrimp research in PWS is documented
by Kimker  and Donaldson (1987), Donaldson  (1989),  Donaldson and
Trowbridge (1989), and Kruse and Murphy (1989).

This project will be carried  out in two general  areas.  One will be
an area of  little apparent impact, the northwestern portion of PWS.
This area  includes  Unakwik  Inlet,  the  site  of previous  ADF&G
research on abundance and growth of spot shrimp.  The second area
will be central  and  southwestern PWS, an area of generally high oil
impact.  This area includes Green Island where ADF&G test fishing
occurred in  1981.   Within each of these  two areas,  fishing will
take place at three  sites.  In the northwestern  sound, test fishing
will occur in Unakwik Inlet,  Port Wells, and Culross Passage.  In
the central and  southwestern sound,  test  fishing will take place
near Herring Bay, Chenega Island,  and Green Island.  An additional
oiled  site will  be  located at Elrington  Passage to increase the
sample  size  for   mixture   modal   analysis   in  1991.     Shrimp

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distribution in these areas has been established by surveying the
commercial fleet.

Fishing will take place at  seven  sites  -  four in oiled areas and
three in unoiled  areas.   Each site will  be  stratified by depth.
Stratum 1 will  be shallow waters - 20 to 70  fathoms.   Stratum 2
will be deep waters - 70 to 120 fathoms.  Based on past research,
spot shrimp are not abundant below those depth ranges.  Because of
the  difficulty of  placing  the  gear at  precise  depths,   it  is
impractical to divide the depth into more  than two strata.  Strata
span 50 fathoms in depth or approximately 65 to 85 fathoms in width
along the bottom  at slopes  of 75 to 100  percent.   Fishing a 100
fathom string will  span the  width of  each strata and allow for a
complete placement of gear over the strata.

Eleven pots spaced 10 fathoms apart will be fished on a long line
so that each string of pots  is 100 fathoms  long.   One 100 fathom
string of gear constitutes a sampling station.  Two stations will
be fished in each stratum at each site for a total of 22 pots per
stratum per site,  or 44 pots per site.  Forty-four pots  is the most
that can be  fished  in a day while  collecting all  of the various
samples  and data.    If  necessary, pots  will  be  redeployed  an
additional day at each site  and  at each depth until a minimum of
500 shrimp are  captured  per depth stratum.    A  total of 264 pots
will be fished during each time period.

Water temperature, salinity,  and dissolved oxygen concentration by
depth will be recorded using a CTD, transferred from the CTD to a
micro-computer and stored on diskette.  CTD  casts  will be at one
station in the deep stratum every day.  The CTD will be lowered at
a rate of 60 meters per minute.   Because  of  the configuration of
the CTD, only readings from the downcast will be used.

Total weight of catch, sub-sample weight, and the weight of each
species in a sub-sample will be  recorded  for each pot on a paper
form at the time the pot  is  retrieved.  The total weight of shrimp
per pot will be determined by weighing the contents  of  each pot on
an electronic scale.  Spot shrimp that are removed as hydrocarbon
samples will be accounted for in the total weight by adding weight
representative  of the number  and size of  shrimp  removed.   The
average number of shrimp per kilogram will be determined. If less
than 500 shrimp are estimated to be contained in all of the pots,
all of the shrimp will be  sampled.  If  the  pots are estimated to
contain more than 500 shrimp,  a  constant  proportion by weight of
each pot will be sampled for a total sample of 500 shrimp.

Each sub-sample will be sorted by species.   Weight and number of
animals will be recorded for each species.  Only spot  shrimp will
be retained for further data collection.  All spot shrimp in the
sub-sample will be measured for carapace length  to the  nearest 0.1
millimeter using  a  digital  caliper and  sex will be determined as
juvenile, male, transitional, or female.  For female spot shrimp,

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egg color  and stage  of development  (eyed  or uneyed);  relative
clutch  size;  presence  of  breeding  dress and  egg parasites  or
parasitic  externa will be  noted.    Each  female  retained  for
fecundity analysis will be identified with a code number to allow
cross reference of fecundity and other data.

Specimens for necropsy  analysis will  be taken  after  the catch is
weighed and processed.   Twenty shrimp from  a single station in each
stratum will be  selected randomly to make up  a  necropsy sample.
Necropsy samples  will  be labeled with the date,  station number,
latitude and  longitude,  sample number,  project leader's  name,
species, and agency.

To prevent  contamination, specimens for hydrocarbon testing will be
taken from the pot immediately after removal from water and before
contents are weighed.   Three spot  shrimp  will  form one composite
sample.  Each composite will be taken from a different pot.   Two
replicates of  the composite will be taken randomly from one station
in the  stratum and the third replicate will come  from the other
station.  Three  samples per site per depth stratum  result in 12
samples per depth stratum  (four sites  X  three  samples)  for the
oiled  area,  and  nine  samples   (3  sites x 3  samples)   per depth
stratum in the unoiled area.  Twenty four  samples (12 samples x 2
depth strata)  will be  taken in  the oiled area and  18  samples (9
samples x 2 depth strata)  in the unoiled area.  This  will allow
hypothesis testing to detect differences in hydrocarbon levels of
1.2 standard deviations with the probability of a type I  or type II
error being 0.05 and 0.10,  respectively.

The number of specimens for one hydrocarbon analysis is dependent
on the size of the specimens collected.  Tissue  volume based on the
average  size  of  the  species  was estimated  and  the   number  of
specimens needed  to provide 15  gm  of  tissue was calculated to be
three spot shrimp. It is estimated that three hydrocarbon samples
from each treatment level are needed for  detecting contamination
between levels.

Twenty five egg-bearing females will be taken at random from each
station to  estimate  fecundity and egg mortality.  A total of 28
stations will yield a total  sample  size  of 700 females.  Specimens
from each station will  be individually  labeled.   Each sample bag
will be labeled with project leader's name,  species name, "eggs",
date,  station, and agency name.

Fecundity  will  be determined  by  removing  the  eggs  from the
pleopods, drying  each  egg  mass  to a  constant  weight,  weighing a
sub-sample of a known number of  eggs,  and expanding the sub-sample
weight to the  weight of  the entire clutch.  Carapace length will be
taken for each specimen at the time of subsampling and assigning a
fecundity number.
                               226

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A minimum number of five shrimp from each station will be sampled
for fecundity  which will allow an adequate  sample  (30 per depth
strata per oil impact level) to test for differences in fecundity
between depth strata and oil impact level.

Objective A will be addressed by estimating the average catch per
pot by weight, sex,  and  species.   ANOVA will be used to test for
significant differences in each of  these categories between strata
(depth), sites,  and oiled  versus  unoiled areas.  To  define the
relationship between hydrocarbon levels and  changes in relative
abundance, statistics for analysis  of covariance  or an appropriate
multivariate technique will be calculated to contrast differences
in hydrocarbon content and relative abundance in  oiled and unoiled
areas.  Changes  in average catch per pot over  time  will also be
analyzed  between different  depth  strata,  sites,  and  oiled and
unoiled areas.

A  size frequency  distribution will be made by sex  to address
Objective  B.    The  hypothesis that   there is  no  significant
difference between strata,  and oil impact levels  for size frequency
distribution will  by tested using quantile-quantile plots,  chi-
square tests or other appropriate methods.  A t-test or a similar
non-parametric test will be used to test for similarity in means.

To meet Objective  C,  the relationship  between size and fecundity
will be examined.   The percentage  of  spot shrimp females bearing
eggs; the stage of spot shrimp  egg  development (color and presence
or absence of eyes); the  percentage of  spot  shrimp egg fouling and
egg mortality; the fecundity by size; and the relative clutch size
will be determined for each station.  Chi-square  tests will be used
to test for differences in  strata,  sites and levels in data which
involve percentages  and  proportions.  Differences between strata,
sites, and impact levels  for fecundity  and relative size of clutch
will be tested for using analysis of variance.

To address Objectives D and E, the average levels of oil present in
spot  shrimp  tissue  by  strata  and  site  will  be  estimated.
Significant differences in hydrocarbon concentrations  between oiled
and unoiled sites  will  be  tested by  analysis of  variance.  To
further define the impact of hydrocarbon levels on the stock, the
percentage of  animals with abnormal tissues in  oiled and unoiled
areas will be  determined.   A chi-square test will be utilized to
test  for  significant differences  in  percentage of  animals  with
abnormal tissues between strata, sites, and  impact levels.
                           BIBLIOGRAPHY

Anderson,  J.W.,  S.L.  Kiesser,  R.M. Bean,  R.G.  Riley,  and B.L.
     Thomas. 1981.  Toxicity of chemically dispersed  oil to shrimp
     exposed to constant and decreasing concentrations in a flowing
     system.  In 1981 Oil spill conference  (prevention, behavior,

                               227

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     control,  cleanup),   Proceedings.  Washington  D.C.  American
     Petroleum Institute, pp. 69-75.

Brodersen, C.C.,  S.D.  Rice, J.W. Short, T.A. Mecklenburg, and J.F.
     Karinen.   1977.    Sensitivity of  larval  and adult  Alaskan
     shrimp  and  crabs to acute exposures  of the  water-soluble
     fraction  of  Cook   Inlet  crude  oil.   In  1977  Oil  spill
     conference    (prevention,    behavior,   control,   cleanup),
     Proceedings.  Washington,  D.C. American Petroleum Institute.
     pp. 575-578.

Brodersen,  C.C.  1987. Rapid narcosis and  delayed  mortality  in
     larvae  of king crabs and  kelp shrimp exposed to  the water-
     soluble fraction  of  crude oil.   Mar. Environ. Res. 22:233-239.

Donaldson, W. 1989. Synopsis of the Montague Strait  experimental
     harvest area 1985 -  1988.  Alaska Department  of Fish and Game,
     Division of  Commercial Fisheries, Regional Information Report
     No. 2C89-04.  21 pp.

Donaldson,  W.  and C. Trowbridge.  1989.    Effects  of  rigid mesh
     panels on escapement of spot shrimp  (Pandalus platyceros) from
     pot gear.   Alaska  Department  of Fish and Game,  Division of
     Commercial Fisheries, Regional Information Report No. 2C89-05.
     22 pp.

Kimker, A.  and W.  Donaldson. 1987. Summary  of 1986  streamer tag
     application  and  overview of  the tagging  project for spot
     shrimp in Prince  William Sound.  Alaska Department  of Fish and
     Game, Division of Commercial  Fisheries,  Prince William Sound
     Management Area Data Report 1987-07.

Kruse,  G.  and   P.  Murphy.  1989.   Summary  of   statewide  shrimp
     workshop held in    Anchorage during  October  24-26,  1988.
     Alaska  Department of Fish and Game,  Division  of Commercial
     Fisheries,  Regional  Information Report No.  5J89.

Mecklenburg, T.A., S.D.  Rice, and J.F. Karinen.  1977. Molting and
     survival  of  king   crab   (Paralithodes  camtschatica)  and
     coonstripe shrimp (Pandalus hypsinotus) larvae exposed to Cook
     Inlet crude oil water-soluble fraction.  In D.A.  Wolfe  (ed.).
     Fate   and   effects   of  petroleum   hydrocarbons  in  marine
     ecosystems  and organisms. Pergamon Press, New York,  NY. pp.
     221-228.

Sanborn,   H.R.   and   D.C.  Malins.   1980.   The   disposition  of
     aromatichydrocarbons   in  adult  spot  shrimp   (Pandalus
     platyceros)  and the  formation of metabolites  of naphthalene in
     adult and larval spot shrimp.  Xenobiotica.   10(3):193-200.
                               228

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Stickle, W.B.,  M.A.  Kapper, T.C.  Shirley,  M.G.  Carls,  and S.D.
     Rice. 1987.   Bioenergetics  and tolerance of the pink shrimp
     (Pandalus borealis) during  long-term exposure to the water-
     soluble fraction and oiled sediment from Cook Inlet crude oil.
     In  W.B.  Vernberg,  A.  Calabrese,  F.P.  Thurberg,  and  F.J.
     Vernberg (eds.). Pollution physiology of  estuarine organisms.
     Belle W.  Baruch  Libr. Mar.  Sci.  17,   Univ.  S. C.  Press,
     Columbia, pp. 87-106.

Vanderhorst,  J.R., C.I. Gibson, and L.J. Moore. 1976. Toxicity of
     No.  2  fuel  oil  to  coonstripe  shrimp.  Mar.   Poll.  Bull.
     7(6):106-108.
                              BUDGET

Personnel Services          $ 35.0
Travel                         1.5
Contractual                   12.0
Supplies                       1.5
Equipment                      0.0

Total                       $ 50.0
                               229

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SUBTIDAL STUDY NUMBER 6

Study Title:   Injury to Demersal Rockfish and Shallow
               Reef Habitats in PWS and Along the Lower KP

Lead Agency:   ADF&G


                          INTRODUCTION

In  light  of  the  findings  of  potential  impacts  on  rockfish
populations continued study of demersal  rockfish populations and
shallow reef habitats is warranted for 1991.  Unlike many species
of  marine  fish,  demersal  rockfish  complexes  are  relatively
sedentary,  residing near rocky  reefs and  boulder  fields.   The
potential impact of the oil spill on various nearshore assemblages
is dependent upon location  of various rockpiles.   The potential
uptake of various contaminants  will be related to the level of oil
contamination  and food web characteristics  of  these reefs.   Of
primary importance are questions  of transport of oil to subsurface
habitats  and  the  potential  for  residual persistence  of  this
contamination.   Khan (1987) reports that  crude oil can contaminate
sediments and persist for long  periods of time in the environment.


Under these conditions, the petroleum  hydrocarbons can  exert a
broad range of effects on animals, from impaired feeding, growth,
reproduction, and changes in behavior; to tissue and organ damage,
damage to blood  cells,  changes in  enzyme activity and changes in
parasite densities  (Khan 1986; Khan 1987; Kiceniuk and Khan 1986;
Rice 1985; Wennekens et al.  1975; Malins et al.  1977; Rice et al.
1977; Gundlach et al. 1983;  Hose et al. 1987; Spies et al. 1982).
These possible affects are especially critical to demersal rockfish
since they  are  long-lived, recruitment  is  low,  and the potential
for long-term stock decline due to chronic exposure to crude oil is
high.  Continuation of this study will  help determine long term
histopathological effects on the  fish and will quantify the extent
to which hydrocarbons persist in the environment.

Only limited baseline data are available for rockfish populations
in PWS and  along the lower Kenai Peninsula (LKP).   Rockfish were
studied as part of a study of nearshore fish assemblages during the
years 1977-1979 in PWS (Rosenthal 1980) and Morrison studied select
reefs along the LKP during 1980 through 1984. These investigations
provided descriptions  of  selected rockfish populations including
estimates of species and prey composition,  density, length and age
composition.
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                            OBJECTIVES

A.   Determine the presence or absence of hydrocarbons in demersal
     rockfish, benthic suspension feeders, and sediments from two
     control and two treatment sites in PWS and two control and two
     treatment sites along the LKP.

B.   Determine  the  physiological  effects  resulting  from  oil
     contamination  through histopathological  examination  of six
     organs,  enzyme activity, and the  examination  of developing
     embryos.

C.   Determine the  feasibility of using otolith microstructure to
     evaluate depressed growth as a result of oil contamination.
                             METHODS

Eight sites (four oiled and four control)  in PWS and along the LKP
will  be  sampled   in   1991.     Demersal  species  of  rockfish,
unconsolidated benthic sediments and sessile  suspension feeders
will  be  collected  at  each  sample  location  for  analysis  of
hydrocarbons.  From the results of these  analyses the mechanism of
hydrocarbon uptake  in  demersal rockfish and the  extent to which
hydrocarbons persist in reef ecosystems may be determined.   The
effects of sublethal hydrocarbon contamination in demersal rockfish
will be  determined through histopathological examination  of six
organs; evaluation  of  enzyme activity;  examination of developing
embryos; and examination of otolith microstructure.  Results will
be compared between oiled and control sites.

Sample sites  will  be  the  same  as  those established in  1990.  A
systematic sampling design will be used to identify sampling sites
within each reef.  Transects will be established at discrete depths
by deploying an anchor  line along specific contours of the reef and
each  end  will  be  marked  by  anchored flag  pole  assemblies.
Coordinates, length, depth, and orientation of the  transect will be
recorded.   The actual  number of sample  sites will  depend on the
length of the transect  and the orientation of  the reef in the ocean
currents.   Sampling will be  conducted during late July and early
August which is the time frame that consistent with 1990 sampling
and also the time  frame that Rosenthal  (1980)  identified as near
the peak  abundance of  rockfish  in nearshore areas.   Collection
methods  for  finfish,   sediment,  and sessile  invertebrates  are
outlined below.

Fifteen  adult  demersal  rockfish  (target  primarily  yelloweye
rockfish Sebastes ruberrimus) will be collected at  each sample site
using  hook  and  line  jigging  techniques. Baited  lures  will  be
lowered to  the substrate and raised enough to allow for adequate
jigging action.  When  a fish is on the  line  it will be retrieved
slowly in order to allow the air bladder  to equilibrate and prevent

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extrusion of the stomach and regurgitation of its contents.  Where
hook and line techniques do not yield results,  divers will verify
the presence or  absence  of  demersal  rockfish assemblages,  and if
present, collect them using spear guns.  Stomach contents will be
collected to  determine composition of  the  prey species  and for
analysis of  hydrocarbons.  Species identification of adult rockfish
will be  accomplished using the  methods of Kramer  and O'Connell
(1988)  and Hart  (1973).

Fifty juvenile demersal rockfish will be collected using variable
mesh, monofilament gillnets set  in the  shallow  areas of the reef
and in  intertidal  zones adjacent to the reefs.   Given estimated
proportions of 0.6  and 0.2 respectively,  sample size was determined
(Zar 1984)  to be  50,  where a =.05.    Species  identification of
juvenile rockfish species will be accomplished using the methods of
Matarese et al.  (1989).

Nine sediment  samples  will be collected at each sample  site by
divers outfitted with  SCUBA equipment prior to  the  collection of
air-lift samples outlined  above.   Each sample  will  be collected
from  the upper two  centimeters of   substrate  and  stored  in
hydrocarbon-free  four   ounce  jars.    Each jar will  be  filled
approximately one-third full.   Excess water will be poured off at
the surface and the sample will be frozen.  Three sediment samples
will be collected at each reef.

Three samples of sessile  filter feeders will be collected from each
reef by divers  outfitted with SCUBA  equipment.   Each sample will
consist of pieces of two or three sessile filter feeders.  Enough
samples will be collected to at least half fill a 4 oz. hydrocarbon
sampling jar.

Samples collected  will  be handled differently  depending upon the
data required and type  of analysis being conducted.  The following
sections explain each type  of preparation that will be  used.  Most
samples collected  will  be  used  for only  one   type  of analysis;
however, each  rockfish captured will  be used   or prepared  for a
variety of purposes.  Rockfish will be processed in the following
specific order:    1)  rockfish will  be measured  to  the nearest
millimeter  (fork  length)  and  weighed  to   the  nearest  gram for
calculation of  condition factor; 2)  tissue will be sampled for
hydrocarbon analysis and histopathological evaluation according to
procedures outlined in proceeding sections; and, 3)  otoliths will
be removed for later age determination.

Length (fork length), to the nearest millimeter,  and weight, to the
nearest  gram,  will  be  used  to  calculate  a relative condition
factor.   Condition factors will  be calculated for  all rockfish
captured.

Ten of the 15 rockfish  (Rice 1990) collected at each reef will be
prepared for hydrocarbon analysis.  All samples will be collected

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from live fish.  Bile samples will be collected first by removing
the whole gall  bladder  and emptying the bile  into  0.5 oz. amber
sampling jars.  Ten  grams  each of stomach,  pyloric caeca, liver,
and muscle  tissue will  be collected from  each rockfish.   Each
tissue type will be stored in separate 4 oz. sampling jars.

Fifteen  live demersal  rockfish,  including  the ten  sampled  for
hydrocarbons, will be collected at each reef  for histopathological
analysis  and processed under  the  guidelines  outlined   by  the
Histopathology  Technical Group  (Meyers 1989).   One centimeter
sections of  tissue  will be removed  from  the  following  organs:
liver, spleen,  kidney,  gills,  gonads, and eyes.   All developing
embryos  will be collected  and preserved  in a  neutral  buffered
formalin solution.

Sagittal otolith pairs will be collected from 50  juvenile yelloweye
rockfish  (measuring  less  than  200  mm)  from  each  reef.   Age
validation  studies  involving  daily  growth  increments,   such  as
Boehlert and Yoklavich  (1987),  typically  utilize  otoliths  from
juveniles  because  growth  is   deposited  more   rapidly,   and
physiological checks and daily  growth  increments are more visible.
Upon collection, otoliths will  be rinsed and stored dry in pairs in
coin envelopes.

Juvenile  otoliths will be  prepared  for  examination  following
methods outlined by Boehlert and Yoklavich (1987).  Otoliths will
be viewed, under transmitted light with a  compound  microscope at
400X  magnification.     Presence  and  location  of  hyaline zones
comprising annuli, daily growth increments,  and checks resulting
from physiological factors including a reduction in growth will be
examined.  The  feasibility of distinguishing  differences in the
type of  zones will be explored by measuring the width of growth
zones deposited over consecutive periods of time (days and years).
Where physiological checks are clearly discernible from annuli, the
presence of  checks  will be determined with respect  to  annuli.
Checks deposited within the growth zone of the previous year will
be noted. The proportion of otoliths containing checks within this
growth zone will be compared between control  and treatment groups.


                          DATA ANALYSIS

Data analysis will consist primarily of the  comparison of results
between control and treatment groups for each of the following:

LeCren's relative  condition factor  (KJ (Anderson  and Gutreuter
1983)  will be calculated for each adult and  juvenile rockfish.  The
mean condition factor for adult and juvenile rockfish for each reef
will be  calculated and  differences  between control and treatment
groups will be tested using ANOVA.
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Rockfish tissues,  sessile  filter feeders, and  sediments  will be
analyzed  for  the  presence  of  hydrocarbons.    Proportions  of
contaminated samples  in each  category  will be  compared  between
control and treatment groups.

For  each  species  the  proportion of  treatment  sites  containing
contaminated samples will be compared to the proportion of control
sites with contaminated samples using a two-sampled z-test from Zar
(1984).

Tissues will be  examined for histopathological  abnormalities and
enzyme  activity  by a  qualified  laboratory.   The  proportion of
samples showing evidence of histopathological abnormalities will be
compared between control and treatment groups for each tissue type
using the z-test from Zar (1984).

Otoliths  from  juvenile demersal rockfish  will  be examined  as
described  in  the  methods  section.     Proportion  of  otoliths
containing checks  between  the last two annuli will  be compared
between control  and treatment groups using the  z-test  from Zar
(1984).  Age composition and mean length-at-age will be calculated
for each species of rockfish.
                           BIBLIOGRAPHY

Anderson, R.O.,  and  S.J.  Gutreuter.   1983.   Length,  weight, and
     associated structural indices.  Chapter 15 In L.A. Neilson and
     D.L. Johnson eds., Fisheries  techniques,  American Fisheries
     Society, Bethesda, Maryland.

Boehlert, G.W. and M.M. Yoklavich.  1987.  Daily growth increments
     in otoliths of juvenile black rockfish, Sebastes melanops: An
     evaluation of autoradiography as a new method of validation.
     Fish. Bull. 85  (4): 826-832.

Chess,  J.R.   1978.     An  airlift  sampling  device for  in  situ
     collecting of biota from rocky substrate.  Mar. Tech. Soc. J.
     12:20-23.

Gundlach, E.R.,  P.D.  Boehm, M. Marchand,  R.M.  Atlas, D.M. Ward, and
     D.A. Wolfe.    1983.    The  fate  of Amoco Cadiz  oil.   Sci.
     221:122-129.

Hart, J.L. 1973.  Pacific  fishes of  Canada.   Bulletin 180,  Fish.
     Res. Board of Can. Ottawa, Ontario, Canada,  pp. 388-453.

Hose, J.E.,  J.N Cross, S.G. Smith and  D.  Deihl.  1987.   Elevated
     circulating   erythrocyte   micronuclei   in   fishes   from
     contaminated sites in California.  Mar. Environ. Res. 22:167-
     176.
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Khan  R.A.    1986.    Effects  of  chronic  exposure to  petroleum
     hydrocarbons  on two  species  of marine  fish  infected  with
     hemoprotozoan, Trypanosoma muranensis.  Can.  J.  Zool. 65:2703-
     2709.

Khan R.A.  1987.   Crude  oil and  parasites in fish.  Parasitology
     Today 3:99-102.

Kiceniuk  J.W.   and  R.A.  Khan.    1986.     Effects  of  petroleum
     hydrocarbons on Atlantic  cod,  Gadus morhua, following chronic
     exposure.   Can. J. Zool.  65:490-494.

Kramer, D.E.  and V.M.  O'Connell.  1988. Guide to northeast Pacific
     rockfishes genera Sebastes  and  Sebastolobus.   University of
     Alaska Marine Advisory Bulletin No.  25.

Malins, D.C.,  E.H.  Gruger,  Jr.,  H.O. Hodgins, N.L.  Karrick,  and
     D.D.  Weber.     1977.     Sublethal   effects   of  petroleum
     hydrocarbons and trace metals, including biotransformations,
     as  reflected  by  morphological,  chemical,  physiological,
     pathological, and behavioral indices.  DCS Energy Assessment
     Program.  Seattle, Washington.

Manen, C. A., Chairperson.  1989.   State/federal damage assessment
     plan. Analytical  Chemistry  Group, National  Marine Fisheries
     Service, Auke Bay, Alaska.

Matarese A.C.,  A.W.  Kendall Jr., D.M.  Blood, and B.V. Vinter. 1989.
     Laboratory guide to early life  history stages of northeast
     Pacific fishes.   NOAA Tech. Rep. NMFS 80.  National Oceanic
     and  Atmospheric  Adm.,  National Marine  Fisheries  Service.
     Seattle, Washington 98115.   625 pp.

Meyers, T.  R.,  Chairperson.  1989.  State/federal damage assessment
     plan.   Histopathology Technical  Group, Alaska Department of
     Fish  and  Game,  Fisheries  Rehabilitation,  Enhancement,  and
     Development Division, Juneau,  Alaska.

Rice, S.D., J.W. Short, and J.P.  Karinen.   1977.   Comparative oil
     toxicity and comparative animal  sensitivity.  In D.A. Wolfe,
     ed.,  Fate and  effects of  petroleum  hydrocarbons  in marine
     organisms and, ecosystems,  proceedings,  Pergamon Press, New
     York.  Pp. 78-94.

Rice, S.D.   1985.  Effects of oil on  fish.  Pp. 157-182.  In F.R.
     Engelhardt ed.  Petroleum effect in  the  arctic environment.
     Pp. 157-182.  Elsevier Applied Science Publishers, London.

Rosenthal, R.J.,  V.  Moran-O'Connell and  M.   C.  Murphy.   1988.
     Feeding ecology  of  ten species  of  rockfishes  (Scorpaenidae)
     from the Gulf of Alaska.   Calif. Fish  and Game 74:16-37.
                               235

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Rosenthal,  R.J.     1980.   Shallow   water  fish  assemblages  in
     northeastern  Gulf of  Alaska:   habitat evaluation,  species
     composition,  abundance,   spatial  distribution  and  trophic
     interaction.  Prepared for NOAA, OCSEAP Program. 84 pp.

Rubin, J.  1988.   A  review  of  petroleum  toxicity and fate in the
     marine environment, with implications for the development of
     a  penalty  table  for  spilled   oil.    Institute for  Marine
     Studies,  University of Washington.   Seattle, Washington.

Spies, R.B., J.S.  Felton,  and L. Dillard.   1982.  Hepatic mixed-
     function oxidases  in California flatfishes are increased in
     contaminated  environments  by oil and  PCB  ingestion.   Mar.
     Biol. 70:117-127.

Wennekens, M.  P., L.  B.  Flagg, L.  Trasky,  D.  C. Burbank,  R.
     Rosenthal,  and  F.  F.  Wright.   1975.   Anatomy and potential
     costs of an oil  spill upon Kachemak Bay. Alaska Department of
     Fish and Game, Habitat Protection Section.  Anchorage, Alaska.

Zar, J.H.  1984.   Biostatistical analysis.   Prentice Hall, Inc.,
     Edgewood Cliffs, New Jersey.
                              BUDGET

Personnel                $   40.9
Travel                        2.7
Services                     63.6
Supplies                     11.8
Equipment                     1.0

Total                    $120.0
                               236

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SUBTIDAL STUDY NUMBER 7

Study Title:   Assessment   of   Oil  Spill  Impacts   on  Fishery
               Resources:  Measurement  of Hydrocarbons  and Their
               Metabolites, and Their Effects

Lead Agency:   NOAA
                         INTRODUCTION

Because petroleum  and its components may cause  severe injury to
fishery resources,  monitoring of the  nearshore fisheries resources
of  PWS.   Such monitoring will include measurement  of petroleum
exposure and short-term effects,  as was done in the summer and fall
of  1989  and the  summer  of  1990.    This  study will  continue to
encompass a  selected  assessment of  long-term biological effects,
including    measurements    of   reproductive   dysfunction   and
histopathological lesions of liver, gill, kidney,  and gonad, as was
done in the summer of 1990 (Varanasi  et  al. 1990,  1991).  However,
the scope of the 1991 study is reduced substantially compared to
studies done in 1989 and 1990, in that the primary study area will
be  limited  to PWS,  and  fewer  species will  be   examined.   This
narrowing of focus  reflects findings of the previous two years, and
is aimed at  continuing only those portions of the  study which are
most likely to assist in documentation  of  injury.   This study will
also include the  measurement of petroleum exposure  and possible
effects in pollock from PWS and the Shelikof Strait.

Certain petroleum  components  [e.g.  AHs]  may  cause reproductive
toxicity and teratogenicity in  rodents  (Shum et  al.  1979; Gulyas
and Mattison 1979; Mattison  and Nightingale 1980).   Similarly,
reproductive impairment has been noted  in  benthic  fish residing in
contaminated areas of San  Francisco Bay (Spies and Rice 1988) and
southern California (Cross and Hose 1988).  Moreover, English sole
from areas of  Puget Sound having high sediment concentrations of
AHs showed inhibited ovarian maturation (Johnson et al. 1988), and
fish from these areas that did mature often failed to spawn after
hormonal treatment to induce  spawning (Casillas et al. 1991).  In
general, reproductive impairment (including reduced plasma levels
of  the  sex  steroid,  estradiol)  was  found in English  sole which
showed  evidence  of  exposure to aromatic compounds.   Moreover,
laboratory studies have shown that plasma levels  of estradiol are
reduced  in  gravid  female   English  sole  exposed  to  chemical
contaminants extracted  from urban sediments  (Stein et al. 1991).
More importantly,  our preliminary laboratory studies  have shown
that exposure to   Prudhoe Bay crude  oil reduced plasma levels of
estradiol in gravid female rock sole.  The continued assessment of
possible reproductive dysfunction in animals from impacted areas
will be very important in  determining biological  damage to living
marine resources as a  result of the EVOS. Histological examination
of ovaries of selected  species  will  be performed to determine if

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ovarian maturation is being affected in animals from oil-impacted
areas.   Fecundity and levels  of  plasma estradiol in  these same
animals  will  be  determined.    Combined  with  measurements  of
petroleum exposure (e.g.  metabolites in  bile and enzyme activities
in liver,  these studies will  allow estimation of the  degree of
reproductive dysfunction which may  be  occurring  in  oil-exposed
fish.

Exposure of animals to crude oil may also result in changes at the
tissue and  cellular  levels  (National Academy  of  Sciences 1985).
Examples of such changes after exposure of fish to oil-contaminated
sediments  include liver  hypertrophy and fatty  liver  in winter
flounder (Payne et al. 1988) and the occurrence of hepatocellular
lipid vacuolization in English sole  (McCain et al. 1978).  Certain
AHs  (e.g.,  benzo[a]pyrene)  are known carcinogens  in  rodents and
fish  (Lutz  1979; Bailey et  al.  1989),  and studies with several
bottomfish  species show that, of  the  xenobiotic chemicals  in
sediments, AHs are most strongly associated with high prevalences
of liver lesions, including neoplasms (Myers et al. 1987; Varanasi
et al. 1987; Baumann 1989).  Generally,  histopathological lesions
of the types noted above  do not  become manifest until at least
several months  after  exposure.   However,  by the  summer of 1991,
fish in and  around oil  impacted sites will  have potentially been
exposed  to  petroleum  components   for    more  than  two  years.
Moreover,  there  are  some  published  data  which  suggest  that
histopathological changes have occurred in some fish species as a
result of exposure to oil  spilled from  the  Exxon  Valdez  (Khan et
al. 1990).

Preliminary  studies  in  1990 suggested that  pollock were being
exposed to petroleum both inside and outside PWS.   This study has
been  expanded  to  cover  assessment of  exposure  and  possible
associated biological effects in pollock,  both inside and outside
PWS.

Briefly,  this study will continue to measure  exposure to oil and
oil components in the biota of PWS and other areas affected by the
oil spill,  by determining levels of hydrocarbon metabolites in bile
and by measuring hepatic AHH activities.  Additionally, the study
will measure a range of biological effects,  especially  indicators
of reproductive dysfunction and histopathological effects.  Only by
employing  such  a  broad spectrum of state-of-the art  chemical,
biochemical and biological methods will analytical  data be  obtained
to document the  degree of exposure and resultant biological effects
of  petroleum   hydrocarbons  on   economically  and  ecologically
important  fish  species.   This information  for important Alaskan
fish species will be incorporated into models for use in  estimating
oil spill impacts on fishery resources.
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                            OBJECTIVES

A.   To  sample selected  fish species  (e.g.  pollock,  yellowfin
     sole, rock  sole,  flathead  sole,  Pacific cod)  from several
     sites inside and outside PWS, with emphasis on  sites inside
     PWS.  Site selection is primarily based on data from the last
     two years of sampling and analyses.  Representative sediment
     samples will also be taken from each benthic sampling site for
     subsequent chemical analysis.

B.   To  estimate  the  exposure  to  petroleum  hydrocarbons  by
     measuring levels of  hydrocarbon metabolites in bile  of the
     above species from oiled and unoiled habitats such to detect
     significant differences in bile concentrations with a = 0.05.
     Additionally, stomach contents of fish showing high levels of
     hydrocarbon  metabolites  in   bile  will  be  analyzed  for
     hydrocarbons,  such  to  detect significant  differences  in
     concentrations with a = 0.05.

C.   To  estimate  the  induction  of  hepatic  aryl  hydrocarbon
     hydroxylase  activity  or  increased  levels  of  cytochrome
     P-450IA1 in the above species from  oiled  and unoiled habitats
     such to detect  statistical  differences  in  levels  of effects
     with a = 0.05.

D.   To estimate the prevalence of pathological conditions in the
     above species from oiled and unoiled habitats such to detect
     statistical differences in levels of effects with a = 0.05.

E.   To  estimate  the levels  of  plasma  estradiol, the  degree of
     ovarian maturation, and fecundity in adult females of two of
     the above species  (yellowfin sole and pollock) from oiled and
     unoiled habitats  such  to detect  statistically significant
     differences with a = 0.05.

F.   To estimate  temporal  changes  in the parameters described in
     Objectives B&C, by comparing data obtained  in  1991 to data
     obtained in 1989 and  1990.  In order to assess either recovery
     or increased damage of habitats from the  oil  spill, trends in
     these parameters  must be statistically  significant at  a =
     0.05.

G.   Using the  above data, as appropriate,  construct  simulation
     models similar to those of Schaaf et. al.  (1987)  for important
     Alaskan fish species for use in estimating oil spill impacts
     on fishery resources.  These models  will incorporate pre-spill
     information  from the fisheries literature on  mortality and
     fecundity   together   with    information  on   reproductive
     impairment, pathological conditions, and biochemical effects
     in fish exposed to petroleum hydrocarbons as a result of the
     spill.
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                             METHODS

A.   General Strategy and Approach

Samples of benthic fish (yellowfin sole, rock sole, flathead sole,
and to a  lesser extent,  Pacific  cod)  will  be  collected from five
sites during 1991, from  mid-May  to  mid-June.   Sites proposed for
sampling  are Olsen Bay,  Rocky Bay,  Snug Harbor,  Sleepy Bay,  and
Squirrel  Bay.     As  feasible,  the  sample  locations  will  be
coordinated with  Subtidal  Study  1.   The selection  of  species is
based  primarily  on  results  obtained  in  1990   and  1989  under
Fish/Shellfish Study 24,  and to a  lesser  extent, Fish/Shellfish
Study 18.  Surficial sediment  samples  for  establishing levels of
petroleum hydrocarbon residues will be  collected  at these sites,
with analyses projected to be done under Subtidal Study 1.  Pollock
will be collected in March, 1990, at several sites inside PWS and
in the Shelikof Strait.  Because of the schooling nature of this
species,   and the  dependence  on assistance  from other federal and
state  groups for use  of  sampling  platforms, sites   cannot  be
predetermined.   Efforts will be made to sample sites representing
a spatial gradient away from the spill's occurrence and path.

Petroleum exposure of fish will primarily be assessed by measuring
(a)  concentrations of metabolites of aromatic petroleum compounds
in  bile,  and  (b)  AHH  activities  in  liver.    These  types  of
measurements are necessary because petroleum hydrocarbons in fish
are rapidly  metabolized  to compounds  that  are not  detectable by
routine chemical analyses.   AHH activity in fish is due primarily
to a single cytochrome,  P-450IA1  (Varanasi  et al.  1986; Buhler and
Williams  1989).   Measurement of hepatic AHH activity will provide
a very sensitive  indicator  of  contaminant exposure of  sampled
animals  (Collier  and  Varanasi 1987; Collier  and  Varanasi 1991).
Moreover, the induction  of AHH activity indicates  not  only that
contaminant exposure has  occurred, but also that biological changes
have  occurred  as a result  of  the  exposure.    In addition  to
measuring  AHH  activity,  cytochrome  P-450IA1 will be directly
quantitated  in   selected   liver   or   tissue  samples   by  an
immunochemical method  recently  developed  at  the  University  of
Bergen (Collier  et al. 1989; Gokseyr 1991).   Direct quantitation of
cytochrome P-450IA1 has  the advantage  of  using  archived samples
frozen at non-cryogenic temperatures  (>  -80°  C) .   Thus future
comparisons may be made between data collected  in  this program and
data from other  sample collection programs,  if samples from the
other  programs   are  subjected  to   the   same   immunochemical
quantitation techniques.

Other biological  effects in fish will  be  estimated by examining
selected  species  for  pathological conditions and  by  assessing
reproductive   impairment  in   suitably   mature    female  fish.
Pathological conditions will include grossly visible abnormalities
(e.g., fin  erosion)  and other lesions  diagnosed  by histological
procedures   (e.g.,   gill    necrosis,    liver   cell    necrosis).

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Reproductive  capacity   will   be  estimated  by   examining  the
developmental stages of ovaries and by measuring plasma levels of
certain reproductive hormones  (Johnson et al. 1988) ,  in addition to
measuring fecundity (Cross and Hose 1988).  The two primary species
for  assessing reproductive   impairment  are  yellowfin  sole  and
pollock.  It  is  anticipated that,  during the respective sampling
periods  (May/June  and March)  ,  these two  species  will  be  at an
appropriate stage in their reproductive cycle for such assessments.
Laboratory studies will also be  conducted to determine the effects
of known doses of oil and oil  components on reproductive processes
in these or related species.

Samples of sediment, and selected stomach contents of fish  (whose
bile had evidence of oil exposure) will be analyzed  (sediment under
Subtidal  Study  1)   for  hydrocarbons  by  recently  developed,
scientifically  sound  and cost-effective  analytical  procedures
involving    high-performance    liquid    chromatography,    gas
chromatography and mass spectroscopy (Krahn et al., 1988).

Environmental  damage  will be   assessed  using  statistical  and
simulation  models,  which  will  be  developed as  part  of  these
proposed studies, as well as from other investigations with related
fish species.  The bile and tissue chemistry data will be used to
establish relationships  between biological  damage  and  estimated
exposures to petroleum hydrocarbons.

B.   Sampling Methods

Sampling activities will be  conducted  at  several  sites  in PWS,
including  unoiled   sites   in  Rocky  Bay   and   Olsen   Bay  and
petroleum-exposed sites  in Snug Harbor, Sleepy Bay and Squirrel
Bay/Fox Farm.  Sample collection will be performed from a charter
vessel for the three flatfish species and cod, at water depths of
approximately 0 to  100 meters.  At each site,  sediment samples will
be collected  with  a  box corer, VanVeen or  Smith-Mclntyre grab.
Sediments will be stored at -  20* C.  The coordinates and depths of
each  station  will  be recorded.   For  pollock,  samples  will be
collected at sites within the oil spill area.

Fish will be collected  with  a  bottom trawl, long-line  gear, or
midwater trawl.   Bottom trawls  will be performed  with  an otter
trawl (7.5m opening,  10.8 m total  length,  3.8 cm-mesh in the body
of the net,  and  0.64  cm-mesh  in the liner  of the cod end).  Tows
will  be of   5  to  15  minutes   duration.    In order  to  reduce
contamination of the catch by free oil, trawling will avoid areas
of surface films or slicks.   If a  net is fouled by subsurface or
bottom oil,  it will  be replaced  (or cleaned, if possible) and a new
area  for  trawling  will  be selected.   Other fish  sampling gear
appropriate to the  species and  conditions  will  also be deployed.
Individuals of  selected target  fish species will  be  sorted and
examined for externally visible  lesions; up to 30 fish of selected
species will  be measured,  weighed,  and necropsied; and tissue

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samples   will   be   excised   and  preserved   in   fixative  for
histopathological examination or frozen for chemical analyses.

C.  Laboratory Analyses

1.   Bile Metabolite Assay (analyses done under Technical Services
     1)

Samples  of  bile  will   be   injected   directly  into  a  liquid
chromatograph and a gradient elution conducted using a Perkin-Elmer
HC-ODS with a gradient of 100% water (containing  5/LtL acetic acid/L)
to  100%  methanol   (Krahn  et  al.  1984,  1986  a,  b,   c) .    Two
fluorescence detectors are used in series.  The excitation/emission
wavelengths  of  one  detector  are  set  to  290/335  nm,  where
metabolites  of  naphthalene  (NPH)  fluoresce.  Excitation/emission
wavelengths  of  the  other detector are  set to  260/380  nm,  where
metabolites of phenanthrene (PHN) fluoresce.  The total integrated
area for each detector is then converted (normalized) to units of
either NPH or PHN that would be necessary to give that integrated
area.

2. Liver Aryl Hydrocarbon Hydroxylase (AHH) Activity and Cytochrome
   P-450IA1 Analysis

Hepatic microsomes are prepared essentially as described by Collier
et al.  (1986) and microsomal protein is measured by the method of
Lowry et al.  (1951), using bovine  serum albumin as the standard.
AHH activity is assayed by  a  modification  of the  method of Van
Cantfort et al.  (1977) as described by Collier et al.  (1986) , using
14C-labeled benzo[a]pyrene  as the primary substrate.   All enzyme
assays will be run under conditions in which the  reaction rates are
in the linear range for both time and protein.  Cytochrome P-450IA1
will  be measured  by  an ELISA  utilizing  rabbit  antibodies  to
cytochrome P-450c isolated from Atlantic cod (Goksoyr 1991).

3.  Histopathology

Histopathological procedures to  be followed are described in the
report  from the  Histopathology Technical  Group  for  Oil  Spill
Assessment Studies in Prince William Sound, Alaska.  Briefly, the
procedures will involve the following: (a)  tissues  preserved  in the
field will be routinely embedded  in paraffin and sectioned at five
microns (Preece 1972);  and (b)  paraffin  sections will be routinely
stained  with Mayer's  hematoxylin  and  eosin,  and for  further
characterization of specific lesions, additional sections will be
stained using  standard special  staining methods  (Thompson 1966;
Preece 1972; Armed Forces  Institute of Pathology 1968).  All  slides
will be examined microscopically without knowledge of where the
fish were captured.   Hepatic  lesions will be classified according
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to the  previously described diagnostic criteria of  Myers  et al.
(1987).  Ovarian lesions will be classified as described in Johnson
et al.  (1988).

4.  Reproductive Indicators

Reproductive activity will be assessed by  examining the ovaries of
the sampled  fish histologically to  determine  their developmental
stage,  and  for  the presence  of ovarian  lesions  that  would  be
indicative of  oocyte resorption (Johnson et  al.  1988).   other
parameters associated with  reproductive   activity  will also  be
measured, including fecundity  (Bagenal and Braum  1971),  plasma
vitellogenin  (Gamst and  Try  1980;  DeVlaming et  al.  1984)  and
estradiol (Sower and Schreck 1982) levels,  and gonadosomatic index
(ovary wt/gutted body wt x 100).   Relationships  between ovarian
maturation,  fecundity, plasma estradiol, plasma vitellogenin, and
petroleum hydrocarbon exposure will then be evaluated.

D.  Quality Assurance and Control Plans

1.  Bile Analytes

Quality assurance procedures for bile analyses  will  include NPH and
PHN calibration standards and  the  calibration standard will  be
analyzed after  every  6 samples and  the RSD will  be reported.  In
addition, one  blank sample  and one  reference material (control
material) will be analyzed daily.  The concentrations of analytes
should be within 2 SD of  the established concentrations in control
material.   Replicate analyses  will be performed  on 10%  of the
samples, if a sufficient amount  exists.

2.  AHH Activity and Cytochrome  P-450IA1

Quality assurance procedures for AHH measurements include duplicate
zero-time and boiled  enzyme  blanks  for each set  of assays.  Each
sample will  be  run in duplicate and  those samples  showing > 20%
absolute  difference  between duplicates  and   >10  units   (pmoles
benzo[a]pyrene metabolized/mg microsomal protein/minute) difference
between  duplicates  will be  repeated.    ELISAs  for cytochrome
P-450IA1  will  be  run  in  triplicate,  and  if  the  resulting
coefficient of variation  (CV)  is > 10%, the outlying replicate will
be omitted from  the calculations. If the CV still exceeds 10%, the
analysis of that sample will be  repeated.

3.  Histopathology

Pathologists  on  this  project  will  use  consistent,  standard
diagnostic criteria  to be strictly adhered to by  those who will
also be examining slides in this project.   These criteria will be
established  using  color  photographs  of  external   lesions  and
standard reference slides containing tissues with the major lesion
types expected in the study.  Unusual or atypical lesions will be

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referred to  specialists for confirmation.   The accuracy  of the
histopathologic diagnosis also will be assured by consulting with
and sending sections of tissues with representative lesion types to
the Registry of Tumors in Lower Animals, National Museum of Natural
History at the Smithsonian Institution in Washington, D.C.

4.  Reproductive Indicators

Quality  assurance  for  the  measurement  of  plasma  estradiol and
vitellogenin include analysis of standards to confirm linearity and
calibrate  the assays.    Blank  analyses will  be   conducted  to
eliminate  matrix  effects.     Analyses  of  pooled  plasma  from
vitellogenic  female English  sole and winter flounder containing
known  levels of estradiol  and vitellogenin  will  also be done.
Duplicate analyses of each sample  to  evaluate performance  of the
assays will also be conducted.  These quality checks  are run daily
with each set of samples.  Fecundity measurements will be done in
triplicate on each individual.
                          DATA ANALYSIS

A.  Statistical Tests

The relative concentrations of  contaminants  in sediment and fish
tissues at the  study sites will be compared statistically using the
Kruskal-Wallis test (ANOVA by  ranks;  see Sokal  and Rohlf 1981, Zar
1984). Where significant differences among chemical concentrations
are  found,  the  a-value  will be understood to  be  <  0.05.   To
determine whether the prevalence of histopathological  effects noted
in each  of the  fish  species  is statistically uniform  among the
sites, the G test for  heterogeneity (Sokal and  Rohlf  1981) will be
performed.

B. Analytical Methods

Where possible, non-parametric statistical tests will be employed
to  avoid assumptions  that the data  are  normally  distributed.
Non-parametric tests give highly reliable results.  The principal
non-parametric  tests  that   will   be  used   are  Spearman  rank
correlation, which has  about  91% of the power of product-moment
correlation when the parametric assumptions are met (Zar  1984) , and
the heterogeneity-G statistic.  Spearman rank correlation will be
used for estimating uptake and metabolism of petroleum hydrocarbons
from oiled  and unoiled habitats when  an independent  measure of
contamination  (e.g.,  levels of AHs in sediment) is available.

The heterogeneity-G statistic (Sokal and Rohlf 1981) will be used
to study prevalence of pathological conditions at oiled and unoiled
habitats.  In addition,  logistic regression (appropriate where the
outcome variable is binomial)  will be used to  model the prevalences
of pathological conditions in relation to contamination.

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The Kruskal-Wallis test  (a  non-parametric  form of ANOVA)  will be
used for supporting statistical analyses of variation in sediment
PAH  levels at  sites  sampled.    If  the  null  hypothesis of  no
differences among sites is rejected at a = 0.05, a non-parametric
multiple comparison test (Dunn 1964; Hollander  and Wolfe 1973; Zar
1984) will be used to  determine  differences  between sites at a =
0.05.  Principal components  analysis  and  LOWESS (Chambers et al.
1983) will also be employed for this purpose; both are methods of
exploratory  data analysis  rather  than inferential  statistical
methods.  Cohen  (1977) will be used for computations of statistical
power.

C. Products

Status reports  will  contain information on  the distribution and
concentrations of petroleum hydrocarbons and their metabolites in
fish tissues and in  sediments  obtained  from  sites in Alaska; the
hepatic activities of AHH and levels of cytochrome P-450IA1 in fish
from sites in Alaska;   and the distribution  and  prevalence of
histopathological  disorders  and  reproductive  impairment  among
selected  species  from those  sites.    Chemistry  data  will  be
submitted in the form of data tables and  distribution maps, and all
data will be stored in computerized data  management programs.  Fish
pathology data will be reported in the form of distribution maps,
tables describing  disease  frequencies of  each species examined,
photographs of gross and microscopic properties of abnormalities,
figures  representing  various  types  of  biological data  (e.g.,
length-weight,  age-weight)  and  discussions   of  the  relative
importance of the types of abnormalities found.   Comparisons of the
characteristics of these abnormalities  will  be made with similar
conditions previously reported in other  marine  areas of the world.
The data management formats were designed in cooperation with the
National Oceanographic Data Center (NODC),  and  are compatible with
the NODC data storage  systems.   In addition, articles describing
the results of  these studies will be published in peer-reviewed
scientific journals.


                           BIBLIOGRAPHY

Armed Forces Institute of Pathology.  1968.  Manual of histologic
     staining methods.  Third Edition (L.G.  Luna, ed.) McGraw-Hill,
     New York, 258 p.

Bagenal, T.B. and E. Braum.   1971.   Eggs and early life history.
     P. 165-198, In W.E.  Ricker,  ed. Methods  for the assessment of
     fish  production  in  fresh  waters.   International  biology
     programme  handbook  3.    Blackwell  Sci.  Pub.    Oxford and
     Edinborough, England.

Bailey  G.S.,  D.E.  Goeger,  and  J.D.  Hendricks.  1989.   Factors
     influencing experimental  carcinogenesis  in laboratory fish

                               245

-------
     models.    P.  253-268,  In  U.   Varanasi  ed.  Metabolism  of
     polycyclic aromatic hydrocarbons in the aquatic environment,
     CRC Uniscience Series, CRC Press, Inc., Boca Raton, FL.

Baumann, P.  C.  1989.  PAH, metabolites, and neoplasia in feral fish
     populations.  P. 269-290,  In U.  Varanasi.  ed.  Metabolism of
     polycyclic aromatic hydrocarbons in the aquatic environment,
     CRC Uniscience Series, CRC Press, Inc., Boca Raton, FL.

Buhler,  D.R.   and  D.E.  Williams.   1989.    Enzymes  involved  in
     metabolism  of  PAH  by  fishes  and  other  aquatic  animals:
     oxidative enzymes  (or Phase I  enzymes).   P. 151-184,  In U.
     Varanasi, ed.  Metabolism of polycyclic aromatic hydrocarbons
     in the aquatic environment. CRC Press, Inc., Boca Raton, FL.

Casillas, E.,  D. Misitano, L.J. Johnson, L.D. Rhodes, T.K. Collier,
     J.E. Stein, B.B. McCain,  and U. Varanasi.  1991. Inducibility
     of spawning and  reproductive success of  female English sole
     (Parophrys  vetulus)  from  urban and nonurban  areas.   Mar.
     Environ.  Res.  (in press).

Chambers, J.  M., W.   S.  Cleveland, B.  Kleiner, and P.  A. Tukey.
     1983.  Graphical methods  for data analysis.   Belmont,  CA.
     Wadsworth International Group.   395 p.

Cohen,   Jacob.    1977.     Statistical  power  analysis  for  the
     behaviorial sciences.  Academic Press,  New York.  474 pp.

Collier, T.K.,  J.E.  Stein, R.J. Wallace  and U. Varanasi.  1986.
     Xenobiotic  metabolizing  enzymes in  spawning  English  sole
     (Parophrys  vetulus)  exposed to  organic-solvent  extracts of
     sediments  from  contaminated  and  reference areas.    Comp.
     Biochem.  and Physiol. 84C:291-298.

Collier, T.K.  and  U. Varanasi.  1987. Biochemical  indicators of
     contaminant exposure  in  flatfish  from Puget Sound,  Wa.  P.
     1544-1549.  In Proceedings Oceans '87 IEEE, Washington, B.C.

Collier,  T.K.  and  U.   Varanasi.  1991.  Hepatic  activities  of
     xenobiotic  metabolizing   enzymes  and  biliary  levels  of
     xenobiotics in  English sole (Parophrys vetulus)  exposed to
     environmental contaminants. Arch.  Environ.  Contam. Toxicol.
     (in press).

Collier,  T.K.,  B.-T.   L.  Eberhart,  and  A.   Goks0yr.  1989.
     Immunochemical quantitation of cytochrome P450  IA1  in benthic
     fish from coastal U.S. waters. Proc. Pac. NW Assoc. Toxicol.
     6:9. (Abstract).

Cross,  J.N. and J.  Hose.  1988.   Evidence for impaired reproduction
     in white  croaker (Genyonemus lineatus) from  contaminated areas
     off Southern California.   Mar. Environ. Res.  24:185-188.

                                246

-------
DeVlaming, V.,  R.  Fitzgerald,  G. Delahunty, J.  J.  Cech,  Jr., K.
     Selman, and M.  Barkley.  1984.  Dynamics of oocyte development
     and related changes in serum estradiol 17-/3, yolk precursor,
     and  lipid  levels  in  the  teleostean  fish,  (Leptocottus
     armatus).Comp. Biochem. Physiol. 77A:599-610.

Dunn,  O.  J.    1964.    Multiple  contrasts  using  rank  sums.
     Technometrics  6:241-252.

Gamst, 0.  and  K. Try,   1980.    Determination  of serum-phosphate
     without deproteinization by ultraviolet spectrophotometry of
     the phosphomolybdic acid complex.  Scand. J.  Clin. Lab Invest.
     40:483-486.

Gokseyr,  A. 1991. An ELISA for monitoring induction of cytochrome
     P-450IA1  in  fish liver samples.   Sci. Total  Environ.   (in
     press).

Gulyas,  B.J.   and  D.R.  Mattison.  1979.   Degeneration of  mouse
     oocytes  in response  to  polycyclic aromatic  hydrocarbons.
     Anat. Rec.  193:863-869.

Hollander, M.,  and D. A. Wolfe.  1973.  Nonparametric statistical
     methods.  New York:  John Wiley.  503 p.

Johnson,   L.J.,  E.   Casillas,  T.K.  Collier,  B.B. McCain,  and U.
     Varanasi.  1988. Contaminant effects on  ovarian development in
     English sole (Parophrys vetulus) from Puget Sound, Washington.
     Can. J. Fish. Aquat Sci. 45:2133-2146.

Khan, R.  A. 1990.  Parasitism in marine fish  after chronic exposure
     to petroleum hydrocarbons in the laboratory and to the Exxon
     Valdez oil spill. Bull.  Environ. Contam. Toxicol. 44:759-763.

Krahn, M.M., M.S.  Myers,  D.G.  Burrows  and  D.C.  Malins.   1984.
     Determination of metabolites of  xenobiotics in bile of  fish
     from polluted waterways.  Xenobiotica.   14:633-646.

Krahn, M.M.,  L.J.   Kittle,  Jr.  and W.D. MacLeod,  Jr.    1986a.
     Evidence  for  oil  spilled  into  the Columbia  River.    Mar.
     Environ. Res.  20:291-298.

Krahn, M.M., L.D. Rhodes, M.S. Myers, L.K. Moore,  W.D. MacLeod, Jr.
     and D.C. Malins.  1986b.   Associations  between metabolites of
     aromatic compounds in bile and occurrence of hepatic lesions
     in  English  sole  (Parophrys  vetulus)  from  Puget  Sound,
     Washington.  Arch. Environ. Contam. Toxicol. 15:61-67.

Krahn, M.M., L.K.  Moore,  and W.D.  MacLeod,  Jr.  1986c.   Standard
     analytical  procedures  of  the   NOAA   National  Analytical
     Facility,   1986:  Metabolites of  aromatic compounds  in  fish
     bile.  Technical memorandum NMFS/F/NWC-102,  25 pp.  (Available

                               247

-------
     from the National Technical  Information  Service of the U.S.
     Department of Commerce,  5285  Port Royal Road, Springfield, VA
     22161).

Krahn, M.M.,  C.A. Wigren, R.W. Pierce, L.K.  Moore, R.G. Bogar, W.D.
     MacLeod, Jr.,  S.-L.  Chan,  and D.W.  Brown. 1988.  Standard
     analytical  procedures  of   the NOAA  National  Analytical
     Facility,  1988:  New  HPLC  cleanup and  revised  extraction
     procedures for  organic  contaminants.   Technical  Memorandum
     NMFS/F/NWC-153, 52 pp. (Available from the National Technical
     Information Service of the U.S. Department of Commerce, 5285
     Port Royal Road, Springfield, VA 22161).

Lowry, O.K.,  N.J.  Rosebrough,  A.L. Farr and  R.J. Randall.  1951.
     Protein measurement with the Folin phenol  reagent, J.  Biol.
     Chem. 193:265-275.

Lutz, W.K.  1979.  In vivo covalent binding  of  organic chemicals to
     DNA  as  a quantitative indicator in the  process of chemical
     carcinogenisis.  Mutat.  Res.   65:289-356.

MacLeod,  W.D.,  Jr.,  D.W.  Brown,  A.J. Friedman,  D.G.  Burrows, O.
     Maynes,  R.W.  Pearce,  C.A. Wigren, and  R.G.  Bogar.   1985.
     Standard analytical procedures of the NOAA National Analytical
     Facility, 1985-1986:   Extractable toxic organic compounds, 2nd
     Ed.    NOAA  Technical  Memorandum   NMFS  F/NWC-92,  121  pp.
     (Available from the  National  Technical Information  Service of
     the  U.S.  Department  of  Commerce,   5285  Port  Royal  Rd.,
     Springfield,  VA  22161;  PB86-147873).

Mattison, D.R.  and M.S. Nightingale.  1980.   The biochemical and
     genetic  characteristics of  murine  ovarian  aryl hydrocarbon
     (benzo[a]pyrene) hydroxylase  activity  and its relationship to
     primordal   oocyte   destruction   by   polycyclic   aromatic
     hydrocarbons. Toxicol. Appl.  Pharmacol.   56:399-408.

McCain, B.B., H.O. Hodgins, W.D. Gronlund, J.W. Hawkes, D.W. Brown,
     M.S. Myers, and  J.H.  Vandermeulen. 1978. Bioavailability of
     crude oil from experimentally oiled sediments to English sole
     (Parophrys vetulus) and pathological  consequences. J.  Fish.
     Res. Board Can. 35:657-664.

Myers,  M.S.,  L.D.  Rhodes  and B.B.  McCain.    1987.    Pathologic
     anatomy  and  patterns  of occurrence  of hepatic   neoplasms,
     putative preneoplastic  lesions  and other idiopathic hepatic
     conditions in  English  sole  (Parophrys   vetulus)  from  Puget
     Sound, Washington,  U.S.A.  J. Natl. Cancer Inst. 78:333-363.

National Academy of Sciences. 1985.  Oil in the  sea;  Inputs, fates
     and effects.  National  Academic Press, Washington, D. C. 601pp.
                               248

-------
Preece,  A.    1972.   A  manual for  histologic technicians.   3rd
     edition.  Little, Brown and Co., Boston, 428 pp.

Schaaf, W.E., D.S.  Peters,  D.S.  Vaughan,  L. Coston-Clements, and
     C.W. Krouse.  1987.   Fish population responses to chronic and
     acute pollution:  The  influence of life history strategies.
     Estuaries 10:  267-275.

Shum, S., N.M. Jensen and D.W.  Nebert.  1979.   The murine Ah locus:
     in utero toxicity and  teratogenisis  associated with genetic
     differences   in   benzo[a]pyrene   metabolism.   Teratology
     20:365-376.

Sokal, R.  and F. Rohlf.   1981.    Biometry.   (Second Ed.)   W.H.
     Freeman and Co., San Francisco, CA, 859 pp.

Sower, S.  A., and  C.  B. Schreck.    1982.    Steroid  and thyroid
     hormones during sexual maturation of coho salmon  (Oncorhynchus
     kisutch)  in seawater  or freshwater.   Gen.  Comp  Endocrin.
     47:42-53.

Spies,  R.B.  and   D.W.   Rice,   Jr.   1988.   Effects  of  organic
     contaminants   on   reproduction   of   the   starry   flounder
     (Platichthys stellatus)  in San Francisco Bay. II. Reproductive
     success of fish captured in San Francisco Bay and spawned in
     the laboratory. Mar. Biol. 98:191-200.

Stein, J.E., T. Horn, H.R. Sanborn, and U.  Varanasi. 1991. Effects
     of  exposure  to  a  contaminated-sediment  extract  on  the
     metabolism and  disposition  of 17/S-estradiol  in English sole
     (Parophrys vetulus). Comp. Biochem. Physiol. (in press).

Van Cantfort, J., J De Graeve, and J.E. Gielen.  1977. Radioactive
     assay for aryl hydrocarbon hydroxylase.  Improved method and
     biological   importance.   Biochem.  Biophys.   Res.   Commun.
     79:505-511.

Varanasi,  T.K.  Collier,  D.E.  Williams and D.R.  Buhler.  1986.
     Hepatic  cytochrome  P-450  isozymes  and  aryl  hydrocarbon
     hydroxylase in English sole (Parophrys vetulus).  Biochem.
     Pharmacol.  35:2967-2971.

Varanasi, U.,  D.W.  Brown, S-L. Chan,  J.T.  Landahl,  B.B. McCain,
     M.S. Myers, M.H. Schiewe, J.E.  Stein,  and  Douglas D. Weber.
     1987.   Etiology of  tumors  in  bottom-dwelling  marine  fish.
     Final   Report   to   the  National  Cancer   Institute   under
     Interagency Agreement YO1 CP 40507.

Varanasi,  U.,  S-L.  Chan, R.C.  Clark, Jr.,  T.K.   Collier,  W.D.
     Gronlund, M.M.  Krahn, J.T. Landahl, and J.E. Stein.  1990. Oil
     Spill   Progress  Report.  Shellfish   and  groundfish   trawl
     assessment outside Prince William Sound.  30 p.

                               249

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Varanasi,  U.,  S-L.  Chan,  R.C.  Clark,  Jr.,  T.K.  Collier,  W.D.
     Gronlund, J.L. Hagen,  L.L.  Johnson, M.M. Krahn, J.T. Landahl,
     and M.S. Myers.  1991.   Oil Spill Progress Report. Shellfish
     and groundfish trawl assessment outside Prince William Sound.
     49 p.

Zar,  J.H.    1984.    Biostatistical analysis.    Prentice-Hall,
     Eaglewood Cliffs, NJ,  620 pp.
                              BUDGET

                           NOAA  ADF&G     TOTAL

Salaries                $122.7  $20.8    $143.5
Travel                    10.5    3.5      14.0
Supplies                  18.9    8.7      27.6
Equipment (disposable)     7.9    7.0      14.9
Vessel support            75.0   40.0     115.0

Total                   $235.0  $80.0    $315.0
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                        TECHNICAL SERVICES

The hydrocarbon  analysis and mapping projects  described  in this
section are designed to provide high quality technical services to
studies described in other portions of the NRDA plan.  Hydrocarbon
analytical  services   includes  the   generation,   archival,  and
retrieval  of  all  chemical  analytical  data.   Mapping  includes
implementing and managing a geographic information system to record
and process data collected by NRDA studies.

Appropriate information on exposure of the resource to hydrocarbon
residues  from the  spill is required to determine  and  quantify
injury.   Detailed  information on the distribution  and  evolving
chemical composition of  the  spilled oil  through time,  in concert
with analyses  of  petroleum hydrocarbons or their  metabolites  in the
tissues of organisms will provide essential  information  to other
NRDA  studies  to  demonstrate  the  relationship  of  injury  to
hydrocarbon exposure.

Samples of water, sediments and tissues for chemical analysis are
being collected by individual studies throughout the entire region
impacted by the  EVOS.   Selected samples are being  analyzed by a
team of participating laboratories in accordance  with a centralized
QA/QC program (Appendix A) which will help ensure that all data are
of known, defensible, and verifiable quality and comparability.

The mapping project continues to develop the  damage  assessment
geographic information system.   The primary data layers have been
collected and verified and large scale production and transmittal
of map products has begun.  Specific data analyses and map product
will  continue to  be  generated   to  support the analytical  and
interpretive needs of NRDA studies.

Although the processing of histopathology samples and information
is  no longer  being supported  by a  separate   technical  service
program,   samples,  analyses, and data continue to  be  generated
within  the  context  of  specific  NRDA  studies.    Oil-induced
histopathological  data  are required by  many  of  the  studies
described  under   Fish/Shellfish,  Birds,  Marine  Mammals,  and
Terrestrial Mammals.   This information continues to be  gathered
under strict quality assurance guidelines  (Appendix B)  by expert
histopathologists   to   ensure   compatibility   of   results  and
evaluations throughout the NRDA program.
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TECHNICAL  SERVICES  STUDY  NUMBER 1

Study  Title:   Hydrocarbon Analytical Support Services and
                Analysis of Distribution and Weathering of
                Spilled Oil

Lead Agency:    FWS, NOAA


                           INTRODUCTION

In order to document the exposure of natural resources in the PWS
and GOA ecosystems to spilled oil,   NRDA projects are collecting
sediment, water  and biota samples  to be analyzed  for petroleum
hydrocarbons.   The  data resulting  from the  analysis of  these
samples is used to define the exposure of that resource to spilled
oil, to indicate the  possible effect of the oil on the resource and
to produce an integrated synthesis of the distribution of the oil
in space and time.  The analytical data must be accurate,  precise
and comparable across projects and throughout the time of the NRDA
process.  To this end, TS 1, a cooperative project between NOAA and
the FWS, coordinates  the chemical analysis of all samples collected
by the NRDA projects.  NOAA manages those samples from federal or
state studies involving  water,  sediment,  fish,  shellfish, marine
mammals - with the exception of sea otters,  and intertidal areas.
FWS manages those samples from studies involving birds, sea otters
and  terrestrial mammals.   Samples  are being  analyzed  at  FWS-
contract   Texas  A&M   University   (TAMU),    and   at   NOAA/NMFS
laboratories.  NOAA  has  lead responsibility for implementing the
Quality Assurance programs, updating and maintaining  the sample
inventory and  analytical databases, and data  interpretation and
synthesis.  FWS bears the main responsibility for Quality Control
of the analytical data and assists in the maintenance of analytical
databases and interpretation and synthesis of data.


                           OBJECTIVES

A.   Measure petroleum hydrocarbons,  hydrocarbon metabolites and
     other   appropriate   chemical/biochemical   indicators   of
     hydrocarbon  exposure  in  the  water,   sediment  and  biota
     collected through the NRDA.

B.   Assist  Project  Leaders  and  field  personnel  in implementing
     appropriate  sample  collection,  identification, shipping and
     chain of custody procedures.

C.   Manage sample tracking and archival.

D.   Oversee a Quality Assurance program to  assure and demonstrate
     the  accuracy,  precision and comparability of all chemical
     analytical data developed by the NRDA.

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E.   Provide analytical data to the Project Leaders in a timely and
     useful fashion.  Assist in the interpretation of these data.

F.   Develop  an  integrated synthesis  of  the distribution  and
     chemical composition of spilled  oil,  as it weathers through
     time,  to  provide  a  detailed  basis  for  final  exposure
     assessment.
                             METHODS

All measures of petroleum hydrocarbons and hydrocarbon metabolites
generated in support of the NRDA are being made in agreement with
the QA/QC plan.  The majority of the samples are being analyzed by
TAMU through a FWS contract.  The remainder  of  the analyses are
being preformed by NOAA/NMFS laboratories.  NOAA and FWS are each
responsible for the analysis costs for their managed samples.

A field manual, "Analytical Chemistry:  Collection and Handling of
Samples" written in  cooperation with  all  of  the Trustee agencies
has been provided  to all identified project  leaders  and used by
NOAA and FWS  in a series of training sessions.  Copies of this
manual and continued training sessions will be available in 1991.

A centralized  sample inventory and  tracking system  utilizing a
customized MS/DOS  R-BASE program resides at  NOAA/NMFS,  Auke Bay
Laboratory.    Each  sample   or  subsample  is assigned   a  unique
identification code, defined in terms  of the material collected or
subsampled and documented to an exact field collection location and
time.  The parent database is updated  and maintained by NOAA.  FWS
provides updated information on their  samples  and archives a read-
only copy of the parent database.

The  quality  of  the  analytical data  developed  for the  NRDA is
assured and demonstrated through the mechanisms described in the QA
plan.  For hydrocarbon analyses,  laboratory performance  is:

     0    assisted through the provision of NIST calibration
          standards, control materials and Standard Reference
          Materials,

     0    monitored through the inspection of the results of the
          analysis of the QC samples (calibration standards,
          blanks, matrix  spikes,  replicates,  SRMs and control
          materials) and

     0    tested  through the  blind  analysis of  accuracy-based
          fully-matrixed  samples.

The program is  similar for those laboratories measuring hydrocarbon
metabolites in bile with the exception that because this  is a semi-
quantitative assay, there are no standards or reference materials.

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NOAA/NMFS, Northwest Center has developed calibration and control
materials and distributes them  to  participating  laboratories for
this measurement.

All analytical data,  bulk  parameters and supporting  QC  data are
archived in a customized MS/DOS R-BASE program at NOAA/NMFS, Auke
Bay Laboratory.   For NOAA  and  FWS-managed samples,  the project
leader receives  all data  in  hard copy  accompanied by  a  simple
summary  sheet indicating  whether  or  not the  sample  contains
petroleum hydrocarbons.   Programming  has been completed to develop
a  series  of  ratios  and  indices indicating  the  quantity  and
composition of the oil in the samples.  All data presently in the
database will be  subjected  to this  review and the results provided
to the Project Leaders.   All data are also available to Project
Leaders in electronic form.

Synthesis has been initiated with TS 3 using the recently completed
ratios and indices indicating the quantity and composition of the
oil in  samples.   It  is anticipated that this  cooperation will
result in a series of maps showing changes in the composition and
concentration of the oil with time. .
                              BUDGET

                         NOAA      FWS          TOTAL

Salaries             $   80.0   $  85.0      $  165.0
Travel                   17.0      10.0          27.0
Contracts             1,868.0     430.0       2,298.0
Supplies                  5.0       5.0          10.0
Equipment                30.0      20.0          50.0

Total                $2,000.0   $ 550.0      $2,550.0
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TECHNICAL SERVICES STUDY NUMBER 3

Study Title:   Implement  and  Manage  a  Geographic  Information
               System (GIS) to Record and Process NRDA Data

Lead Agency:   DNR and FWS

Cooperating Agency:  DEC


                           INTRODUCTION

The purpose of Technical Services No. 3 (TS 3) remains unchanged:
the group is charged with implementing and managing the geographic
information system  (GIS)  to  record  and  process data collected in
NRDA  studies.    Primary data  layers  have  been  collected  and
verified.  Additionally,  TS 3 has  begun  large  scale production and
transmittal of NRDA map products.
                            OBJECTIVES

A.   Produce  and disseminate  maps and  analytical products  for
     participants in the NRDA process.

B.   Create and maintain, throughout the NRDA process, a database
     pertinent to the overall damage assessment process, which is
     accessible to all participating agencies.
                             METHODS

Methods are  the same  as  described in the  1990 study plan.   In
addition to the data layers described  in the 1990 study plan, data
layers have  been or will  be prepared for study  site locations,
sampling  locations,  beach  segment locations  and  multi-thematic
atlases of pre-spill data  from various sources.  Additional data
layers will  be added as needed by investigators  and the Trustee
Council to enable geographic-based  compilation of study results and
other pertinent data.

                              BUDGET

                              DNR         FWS     TOTAL

Salaries                 $  434.6     $ 185.0   $ 619.6
Travel                        8.6         6.0      14.6
Contracts                    84.7        16.0     100.7
Supplies                     52.4        10.0      62.4
Equipment                    76.0        83.0     159.0

Total                    $  656.3     $ 300.0   $ 956.3

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DETERMINATION OF INJURY TO CULTURAL RESOURCES

Lead Agencies:  USFS and DNR

Cooperating Agencies:  ADF&G, FWS, NFS


                           INTRODUCTION

Holocene  richness  and  diversity  of  resources  resulted in  the
development of the  largest prehistoric populations in Alaska along
the Pacific  mainland and  island  coasts.   Kodiak Island  had  the
largest, most dense prehistoric population of Eskimo peoples in the
world.   Similar ecological  abundance  suggests PWS  and mainland
coasts also supported major human populations.  The region of oiled
beaches includes large  areas  where few archaeological surveys have
been done.  To determine injury,  specific information is needed on
the location, number, and  character of  historic sites within the
EVOS area.   This information is  obtainable through  intensive on-
the-ground sample surveys and direct testing.


                           OBJECTIVES

This study includes  activities designed to identify and quantify
injury to cultural resources from a scientific standpoint and to
develop  the  foundation for  a  meaningful program to  restore  and
rehabilitate  archaeological  resources.   To  determine  the injury
caused by the spill, the study will focus on the following:

A.   Impacts on soil chemistry (pH, calcium, phosphate);

B.   Impacts  on soil   structure  and   inclusions  (stratigraphy;
     charcoal);

C.   Impacts  on artifacts  including   petroglyphs,  bone,  wood,
     ceramic, fiber and shell;

D.   Impacts  on vegetative   cover of  sites,   including new  or
     increased erosion on the sites;

E.   Occurrence of theft  or  vandalism  on sites, including new or
     increased incidences.
                             METHODS

1.   Activities will be performed in a manner consistent with the
     Secretary  of the  Interior's Standards  and  Guidelines for
     Archaeology  and  Historic Preservation  (48  Fed.  Reg. 44716-
     44740, September 29, 1983).


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2.   Through a literature search and in-field surveys, an estimate
     of  the number,  type,  character,  and  the significance  of
     archaeological sites  in the area affected by  the  oil spill
     will be determined.

3.   Develop  typologies based  on site  type,  time period,  and
     location.

4.   Using  the  typologies  developed,  a representative  sample of
     archaeological site types and locations to be investigated for
     impacts, will be selected.  The sample will include sites in
     unoiled areas to serve as control sites.

5.   Conduct archaeological  investigations  at  the  selected sites
     and locations.

6.   Oil spill  response  workers  and government employees will be
     interviewed concerning impacts to archaeological resources.

7.   A  laboratory analysis  of  the  effects  of the  oil  on  the
     physical characteristics of  the soil column will be performed.
     Attention will be given to  its component parts to determine
     changes in preservation, soil compaction,  stratification, and
     obscuration of the stratigraphy,  as well as leaching and the
     chemical breakdown of organic materials.

8.   Radiocarbon age determinations and  soil  sample analyses for
     pH, calcium, and phosphate will be performed.

9.   Pre-  and  post-spill  vandalism  and  erosion   data will  be
     compiled  and evaluated  to  establish  rates  and effects  of
     vandalism and erosion.
                            DISCUSSION

To assess the potential  injury  to archaeological sites along the
coast, three physical zones can be established:  submerged (below
the lowest low tide),  intertidal  (between the lowest low and the
highest high tides), and shore  margin uplands (above the highest
high  tide).   The  greatest potential  for injury  exists through
direct  deposition  of  oil  in  the  intertidal  zone.    Secondary
transport into adjacent submerged areas and uplands may also injure
archaeological sites. Upland archaeological sites are also subject
to contamination  from  transportation of oil  by  wind,  storm tide
inundation, migration of contaminants in ground water,  oiled bird
and  animal movement  from  the  feeding/travel  corridor of  the
intertidal   zone,    and   their   death   and  decomposition   on
archaeological deposits.   Theft  of  artifacts and  vandalism to
archaeological resources  are  potential  dangers in the intertidal
and upland  zones.    The  intertidal  zone  contains archaeological
sites of great variety, numbers, and  susceptibility to oil injury.

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Shipwrecks,  eroded/scattered  artifacts,   inundated  stratified
archaeological deposits,  prehistoric  rock art,  prehistoric fish
weirs, and remnants of structures or objects deliberately placed in
the intertidal zone are among the site types known to exist.  The
shore margin uplands may contain all the previously mentioned site
types, plus  burials,  above-ground  structures,  and  recognizable
resource collection locations such as culturally modified trees.

In  the two  higher elevation  zones,  a  major  potential  injury
resulting from oil contamination is interference with traditional
archaeological dating techniques.   Radiocarbon dating depends on
comparison of the ratio  of radioactive  carbon  14 to  carbon 12 in
the sample being  analyzed.   Because  petroleum contains  abundant
radioactively-inert carbon  from organisms  dead for  millions of
years, and the use of radiocarbon dating for  dates  up to 35,000
years ago, contamination by even a small amount of ancient carbon
is expected to result  in age determinations that are significantly
older  than the  archaeological event  being dated.    This  would
seriously compromise radiocarbon dating as a technique for dating
human activities  and paleoenvironmental events and conditions.  The
potential for affecting age determination may be significant even
in  areas  where only  a  sheen  exists  and may  be  investigated in
assessing  injury.   In  cases  of  oil  contamination  in stratified
archaeological deposits, masking of the visibility and alteration
of  the chemical  components  of the microstratigraphy may   also
affect archaeologists' ability to trace strata.

Both direct and indirect injuries to archaeological sites may have
occurred from response  and treatment  activities,  as  well as from
increased  activities  in the resource areas.   Further, increased
access  of personnel  to remote  areas  may have  increased  the
knowledge  of  site  locations  and  potentially  may  accelerate
vandalism,  theft  of  heritage  resources,   and  damage  to  the
scientific value of the sites.

Field study activities did not  occur in 1990,  but will be performed
during the  1991  field season.   Funds to perform the study were
obligated but not spent.  The  budget described below reflects the
cost of including additional study sites and the anticipated cost
of completing follow-up work once data are received.

                              BUDGET

                         USFS      DNR       Total

Personnel            $   85.0   $157.3      $242.3
Travel                   18.0      6.3         24.3
Contracts                 0.0    522.4       522.4
Supplies                  0.0      1.6         1.6
Equipment                 0.0      1.0         1.0

TOTAL               $    103.0   $688.6      $791.6

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PART II:  PEER REVIEWERS/CHIEF SCIENTIST

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^

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SCIENTIFIC PEER REVIEWS/CHIEF SCIENTIST

Lead Agency:   DOJ, DOA, DOI, NOAA


                           INTRODUCTION

Acceptable  scientific procedures  contemplate  a  process  through
which study plans,  methodologies  and data supporting conclusions
are  subjected  to  objective,  rigorous  review  by  peers.    The
government  has identified  a number of  biologists,  ecologists,
chemists and statisticians to perform this function in connection
with the natural  resource  damage  assessment studies described in
this plan.   These  scientists also  may  serve the  government as
expert witnesses, testifying regarding damages resulting from the
EVOS.


                            OBJECTIVES

A.   Ensure that the government's  damage assessment studies follow
     acceptable science procedures  and produce valid conclusions
     supported by accurate data.

B.   Produce an integrated assessment of the damages resulting from
     the EVOS based on  the  many individual and disparate science
     and economic analyses.


                             METHODS

A Chief Scientist will be charged with coordination and direction
of all scientific  damage assessment studies,  including synthesis
and peer review efforts. Certain  of the peer reviewers will focus
on the primary areas of scientific damage assessment,  i.e., coastal
habitat,  marine  mammals,   birds,   fish,  shellfish,  terrestrial
mammals, air  and water, subtidal areas  and  archaeologic sites.
Others,  ecologists and biostatisticians, will compare and link data
and findings among the groups.


                              BUDGET

The  federal  trustee  agencies will  reimburse  the  Department of
Justice and NOAA in equal shares.

Department of Agriculture                             $772.0
Department of Interior                                 772.0
National Oceanic and Atmospheric Administration        772.0

Total                                               $2,316.0


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FART III:  ECONOMICS

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                         ECONOMIC  STUDIES

The studies in this section are federal studies designed to assess
the economic value of injury to natural resources associated with
the EVOS.   The following study descriptions  are  very similar to
those for  1990 because the  studies  are ongoing.   An additional
study, estimating  the economic damages  to consumers of petroleum
products, may be initiated if a relationship between the EVOS and
the observed petroleum market price increases can be established.
State studies designed to  assess  the  economic value of injury to
natural resources resulting from the EVOS are not discussed in this
document.   Litigation concerns continue  to prevent disclosure of
detailed progress to date and preliminary results.

The  federal  studies  cover  eight  major  areas:   (1)  commercial
fishing, (2) public land values,  (3) recreation,   (4) subsistence,
(5) intrinsic  values,  (6)  research programs,  (7) archaeological
resources and (8) petroleum price impacts.
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ECONOMICS STUDY NUMBER 1

Study Title:  Commercial Fisheries Losses Caused by the EVOS


                           INTRODUCTION

This study will continue to build upon the results of the previous
years' efforts.

The  EVOS  may have  resulted  in  substantially  reduced  seafood
production  at several ports  including  Cordova,  Seward,  Kodiak,
Kenai, and Homer, which are some of the most important commercial
fishing  ports in the  United States.   Both  short-term impacts,
through closure of certain fisheries, and long-term effects, such
as  reductions in population  that will  not become  apparent  for
several years, may occur.  These impacts  may affect both the supply
of and demand for seafood.

For example, changes in quality  (both real and perceived) may have
occurred, which  could  adversely affect  seafood markets.   In  the
case of  several  important  commercial salmon fisheries,  the spill
resulted  in harvests  being confined to  "terminal"  areas,  thus
restricting traditional fishing patterns and timing of the harvest.

Terminal  area harvests occur in close proximity  to the salmon's
spawning  grounds.   The result can be a  significant reduction in
quality,  as  compared  to   salmon  harvested  in  more  typical
circumstances, i.e., more distant from,  but en route to, spawning
sites.   The reduction  in quality may affect the salmon's overall
marketability  and/or  its  appropriateness  and acceptability  for
specific product forms.  In either case,  seafood consumers at every
market level incur losses.

Salmon is one of  several commercial  species group which may have
been  adversely  affected.   Others may  include Pacific herring,
shellfish, and groundfish.


                            OBJECTIVES

Measure the economic loss to seafood  consumers caused by the EVOS.


                             METHODS

The investigators are  in the process of determining which species
were  injured  by the  spill.     Conceptual  models  of  consumer
preferences and market  characteristics for certain seafood products
are  being  developed.    A  methodology   to assess  statistically
significant changes  in the level and quality of  harvest is also
under  development.     Data  collection  and analyses  will  also

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continue.   The models will  be used to  estimate the  demand for
various  seafood products,  the price changes associated  with the
spill/ and the effects of seafood quality and quantity changes on
consumers.
                             BUDGET

Total:                        $265.5
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ECONOMICS STUDY NUMBER 4

Study Title:  Effects of the EVOS on the Value of Public Land


                           INTRODUCTION

The EVOS affected  subtidal,  intertidal,  and uplands areas on the
shore of PWS and the GOA.  This study will assess the lost market
value of publicly  held lands attributable to the  oil  spill.   It
will estimate market  demand for leases and  sales  of  land in the
impacted areas,  and project changes in total value of public lands.


                            OBJECTIVES

Determine the change in market values of public lands.


                             METHODS

Land appraisals are a common method of assessing the market value
of land.   Appraisers usually  estimate the market  value  of land
parcels from the selling price of  similar parcels.   Because no two
parcels  are  identical,  adjustments  are  required  to  achieve
comparability.   For the purposes  of appraisal, market  value is
generally defined  as  the amount in cash,  or in terms reasonably
equivalent to cash,  for which, in all  probability, the property
obligated to  sell  to a knowledgeable purchaser, who  desires the
property but  is  not obligated to buy.   Using this definition of
market value, the  effect of  the oil  spill  on land  values will be
estimated as the  difference between the pre- and post-spill selling
prices.


                              BUDGET

At present,  no additional funds have been requested  for this study.
There may not have been sufficient land transactions to employ as
the basis for determining any changes in the value  of public lands
affected by the  spill.    If  it  is determined  that  there were
adequate land sales  to support  and  economic  valuation  of  the
impacted lands,  then this study will be continued and funded with
the amount needed  to determine the extent  of the  lost values to
public lands.
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ECONOMICS STUDY NUMBER 5

Study Title:  Economic Damages to Recreation


                           INTRODUCTION

This study will continue to build upon the results of the previous
years' efforts.

The EVOS has impacted natural resources that support a wide range
of recreational  activities including fishing,  hunting,  boating,
hiking,  camping,  and  sightseeing.    Because  of  their  unique
attributes, these resources attract recreationists from throughout
the United States and other countries to PWS and the GOA coast.

The  EVOS  may  result  in  economic  damage  to  those  resources'
recreational services in two principal ways:  1) some recreationists
who otherwise  would  have gone to  the area  choose   a substitute
activity  and/or  area, thereby  potentially suffering a  loss  in
personal satisfaction and possibly incurring increased costs; and
2)    recreationists  who  visit  the  area  may  suffer  reduced
satisfaction  because  of  the oil   spill's  adverse  impacts  on
recreational services that  the natural  resources  otherwise would
have provided.   These types of losses may have been experienced by
sea kayakers, users of charterboat services/recreational fishers,
users of  air charters,  hunters,  cruise ship patrons  and general
tourists.

While  relatively  few in  number,  sea kayakers  may have  been
significantly affected by the oil  spill.  Kayaking trips are taken
from Valdez,  Kodiak, Homer,  Whittier  and  Seward to  the western
portion of PWS and the bays along the Kenai peninsula and Kodiak
Island.  A typical trip involves charter boat transportation to a
site some distance from port.  Most  trips  last  more than one day
and   thus   include   both  kayaking  and   wilderness   camping.
Southcentral Alaska includes some  of  the premier kayaking areas in
the world.

The potential  effect  of  the oil spill on kayakers could  take
several forms:

      beaches used for wilderness camping are oiled and unusable;
      wilderness  scenery is despoiled and sense of  pristine
      environment is lost;
   -  wildlife viewing opportunities are reduced;
      unoiled areas suffer from increased congestion;
      clean-up activities  make boats for transport  expensive or
      impossible to charter;  and
      clean-up activities  spoil the  wilderness  nature of the
      experience.


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All of these potential  effects  may have occurred during the 1989
season and in subsequent years.

Recreational activities that use the services of charterboats and
other private boats  for hire  are typically less intense than sea
kayaking,  but  are   far  more  numerous.    Vessels  for hire  and
charterboats range from the standard six passenger charterboats to
large tour boats carrying over a hundred passengers.  All types of
vessels  for  hire have  been impacted by  cleanup activity.   For
brevity  in  this proposal,  this entire group  is referred  to  as
"charterboats".    Charterboat  related  recreational  activities
include salmon and halibut fishing,  sightseeing and viewing marine
wildlife and ferrying for  wilderness  camping in the PWS,  KP,  and
Kodiak areas.   Charterboats  go out of Valdez,  Whittier,  Homer,
Kodiak, Seward and the smaller villages in southcentral Alaska.

Because  access  to  the general  area  is  not   easy,  there  are
potentially substantial  impacts which can be measured through a
careful study of the charter  fleet.   The  purpose of such a study
would  be to  determine  the  reduction  in the  use  of  the  PWS
environment through the charter fleet as a consequence of the oil
spill.

The  level of  participation  in recreational  fishing among  the
residents of Alaska is  far greater than among the residents of any
other  state  in the  United States.  Marine  recreational  fishing
originates in all  major towns on the PWS  as well  as Cook Inlet,
Kodiak Island and the KP  and  the AP.   Fishing trips are taken in
several ways  -  from shore,  from private boats  and  from charter
vessels.   Because access by car from Anchorage is relatively easy,
shore  fishing  and  private boat  fishing  on the Kenai  is  quite
popular.   All kinds  of  fishing draw large numbers of tourists to
Alaska.

The study of charterboats will address only part of the potential
recreational fishing effects.   It is  possible  that the oil spill
had detrimental  effects on  shore and private  boat recreational
fishing,  as well.  For example,

a)   fishing trips in the potentially oiled areas may have declined
     due to fear of contaminated fish and waters;
b)   anglers may not have been able to find accommodations in areas
     where  they  wanted  to  fish  because  of  cleanup  related
     activities;
c)   the value of particular fishing trips out of the potentially
     oiled  zones may  have  declined  because  sites  became  more
     congested.

Each season,  a number of cruise ships pass through PWS on their way
from  Seattle  or Juneau to Whittier  where they  discharge their
passengers for the train  trip to Anchorage.   The likelihood that
these individuals were directly affected by the oil spill is small,

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but many have  canceled  their trips because of fear that  the oil
spill would spoil the experience.

The general tourist activity  sub-component of the proposal differs
from the others  in that it  is  not directed toward one  specific
recreational  activity.    Here  the goal  is to  determine,  from
aggregate level data,  the extent to which general tourist activity
in the area of  the spill may have been dislocated because of clean-
up activities.  There will have been losses to recreationists if
these  activities  were  diverted away  from areas  thought to  be
contaminated by the spill or  affected  by the congestion  and lost
services associated with clean up.   Some of the marine related part
of  this damage  will be  captured  in  the  investigation  of  the
charterboats and kayaking.   However, those people who  do not plan
to use boats but rather state parks or other facilities  will not
have been covered.
                           OBJECTIVES

Develop estimates of economic damages to recreationists.


                             METHODS

The study  will continue  to  look at  the impact  of the  EVOS  on
various consumptive and nonconsumptive recreational activities.

Sea kayaking:   This  study contains  several  stages:     (1)  the
relevant sea kayaking population will be  identified;  (2) a survey
instrument which will contribute to  both recreational  demand and
simple  contingent  valuation analysis  will be  created;  (3)  the
survey  instrument  will be pre-tested;    (4)  the survey  will  be
administered; and (5) the survey results will be analyzed.

Charterboat activities:  Data for this study will  also be collected
through a survey.  After development of a  theoretical framework for
damage measurement,  the sample size will be defined.  A survey will
be designed  to determine  the  periodic  recreational and cleanup
activities  undertaken by each  charter  vessel, the  number  of
recreationists served, the extent of cancellations and the amount
of time the vessel  was  involved in  clean  up activity.   Vessel
owners may also be interviewed in person.  Finally, the data will
be analyzed.

Recreational fishing:  There  is  an existing model for recreational
fishing  in the  KP area.   This  model  will  be  investigated  to
determine its applicability to the EVOS.

Cruise  ship tours:    Cruise ship  firms  will  be contacted  to
determine whether demand for  cruise  ship  tours to PWS was affected
by the  EVOS.   If there is evidence  of  substantial reductions in

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demand, methods of estimating the actual losses to recreationists
will be explored.

General  tourist activity:   Assuming  that  aggregate  effects  on
tourism may be accurately estimated, this study will compare those
aggregate  effects  with  the results  of  the  activity-directed
substudies to determine whether important categories of  losses have
been missed.

Additional  substudies:   The recreational  losses  study may  be
revised to include economic analysis of the impacts of  the EVOS on
other  recreational activities  such  as  hunting and  use of  air
charters to gain access to areas used for recreation.
                              BUDGET

Total:                     $  390.4
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ECONOMICS STUDY NUMBER 6

Study Title:  Losses to Subsistence Households


                           INTRODUCTION

This study will continue to build upon the results of the previous
years' efforts.

Several communities  on the  shores of PWS,  LCI,  KAP  are highly
dependent upon noncommercial  fishing,  intertidal  food gathering,
marine mammal  hunting, and  land  mammal hunting  for subsistence
uses.   Among  the small  subsistence  communities are  Tatitlek,
Chenega Bay, English Bay, Port Graham,  Ouzinkie, Port Lions, Larsen
Bay,  Karluk,  Akhiok,   Old  Harbor,  and  Chignik  Bay.    Larger
subsistence  communities include Cordova,  Valdez, Seldovia,  and
Kodiak.  Subsistence uses  are defined  as  rural Alaska residents'
customary and  traditional  uses of wild, renewable resources for
direct personal  or family consumption as  food,  shelter,  fuel,
clothing,  tools, or transportation; for the making and selling of
handicraft  articles  out   of  nonedible byproducts   of  fish  and
wildlife resources taken for  personal  or  family consumption; for
barter, or  sharing for personal  or family consumption;  and for
customary trade.  Those uses are designated as the priority public
consumptive use of wild resources.

Following the  EVOS, subsistence harvests were reduced in several
communities.   This  could  have  important  ramifications  in  the
economy and social order of the communities.  Potentially important
economic  losses  to  the communities  include:    (1)  subsistence
losses;     (2)   local   inflation   affecting  harvests  and  food
procurement;   (3) damage to subsistence property;  and (4) loss of
intrinsic value to subsistence users.
                            OBJECTIVES

A.   Conduct a literature review and compile base-line information.

B.   Document  the  extent  of oil contact and  clean-up  on or near
     historic harvest sites.

C.   Document  the  changes  in  subsistence use through time  (i.e.,
     species selection; harvest timing,  quantities,  areas, methods,
     and efficiency; and household participation rates  in harvest,
     use, sharing, barter, and exchange).

D.   Document  local  social  and  economic  changes  that   affect
     subsistence use,  including wage/labor patterns, income levels,
     inflation  rates in  the villages  for  goods and services,
     cleanup work, outside demands, and  industry demands.

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E.   Assign monetary values to losses to subsistence households.

                             METHODS

Field  observations  and  interviews  will  be  used  to  collect
information.  Changes in subsistence use and socioeconomic patterns
will be determined by conducting systematic household surveys and
interviews,  and comparing  these data  to historic  information.
Where applicable, market prices and price imputation will be used
to estimate damages.  For marketed goods, the  cost of replacing the
goods injured by the spill will normally be the measure of economic
damage.  However, the adverse effects of the spill extended beyond
marketed goods.   A number of methodologies are being considered for
the  estimation  of economic  damages   to  non-market  goods  and
services.

                              BUDGET

Total:                      $ 532.1
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ECONOMICS STUDY NUMBER 7

Study Title:   Total Value  of Natural  Resources  Injured  by  the
               EVOS
                           INTRODUCTION

This study was formerly titled "Loss of Intrinsic Values Due to the
EVOS."  The study title has been  changed  to  reflect the scope of
this study more accurately.  This  study will assess both use and
intrinsic values of the  injured natural resources.  The study will
continue to build upon the results  of the previous years' efforts.

Intrinsic values include existence value, option value, and bequest
value.  These  values are independent of the economic values arising
from direct use of natural resources  and cannot be  measured by
observing use  of  the  area affected by the EVOS.   Resources with
intrinsic values include  fish, birds  and  mammals,  along with the
wilderness character,  ecological integrity and/or scenic quality of
certain areas.  These values are only imperfectly captured by the
prices of goods traded in markets.   Accordingly, non-market methods
must be used to calculate  intrinsic values. This study is designed
to use  the  contingent valuation method to determine  the loss in
both intrinsic and use values resulting from the oil spill.


                           OBJECTIVES

Determine the  loss in  the value of natural  resources  injured by the
EVOS.
                             METHODS

The  contingent  valuation  method  involves  use  of  surveys  to
determine the values that people place on goods.  This study will
require  development  of  a  conceptual  framework  for  contingent
valuation survey  design  and analysis  of survey results.   Next,
research will be  conducted  to  determine the  most  accurate survey
instrument  for  assessing intrinsic  values.   This  research will
involve  consultation  with economists and survey  design experts.
Substantial preliminary testing of survey formats will be conducted
among small groups of people to verify the accuracy of the survey
instrument.    A  nationwide survey  will be conducted using  a
professional survey research firm.   Econometric analysis will be
used to interpret the results of the survey.

                             BUDGET

Total:                      $1,964.6
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ECONOMICS STUDY NUMBER 8

Study Title:   Economic Damage  Assessment of Injury  to Research
               Programs Affected by the EVOS
                           INTRODUCTION

The EVOS affected research programs in the vicinity of the spill,
resulting in damage  to or loss of various  research and resource
monitoring  studies.    Opportunities  to  study  natural  resource
systems in the affected area may have been lost or diminished as a
result of the EVOS.  Research studies underway before the spill and
conducted, permitted, cooperatively participated in, sponsored or
funded by the federal government likely were impacted.  One example
is a study  involving  tagging of fish that  was  in  progress in an
affected area of PWS.  Determination of the set of studies affected
and the extent or degree of damage will require careful evaluation
and study.
                            OBJECTIVES

Assess damage to and economic loss of research investigations, and
account for  the cost of resources expended  in  affected studies,
focusing on research-based expenditures made or committed to before
the oil spill.
                             METHODS

The first step in this study is to identify the universe of studies
that were underway in the affected area at the time of the spill.
The  next step  requires a  determination  of  which  studies  were
negatively impacted by the spill.  Some of those impacts may have
been so significant that the entire study  was discontinued.  Other
studies may have been able  to continue,  but only at an increased
cost caused by the impacts  of the spill.   For example, sample sets
may have been destroyed  or the study may have been moved to another
area.    Once  the  universe  of  affected research  programs  is
identified,  this  study will  value  the  destroyed  and  damaged
research studies  by looking first to  total  project  costs,  extra
funds  expended and amounts  spent on  each study prior  to  being
impacted by the spill.

                              BUDGET

Total:                       $104.9
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ECONOMICS STUDY NUMBER 9

Study Title:  Quantification of Damage to Archaeological Resources


                           INTRODUCTION

Archaeological sites along the many miles  of  oiled coastline and
intertidal zones may have been physically injured by oil.  Upland
sites may have  been injured  by erosion caused by  destruction of
site vegetation  or transportation of  the oil  inland.    Loss to
archaeological  resources  includes  direct and  indirect  oiling.
Determination of the number of cultural resources impacted by the
oil  spill   as  well  as  the  type  and  extent  of  injury to  the
archaeological sites has been moved to  a  separate  science study.
The  economics  study is  now  limited to  quantifying the  loss to
archaeological resources.


                           OBJECTIVES

Assess the economic damages to archaeological sites.


                             METHODS

The archaeological science  study will create a database containing
listings of the  oil impacted  areas and a model for the kinds of
cultural resources  impacted,  the  degree of  the  impact  and the
physical setting of the injured resource.  Both use and intrinsic
values of archaeological resources may have been impacted.

Use Value

1.   Effects  of  the  scientific  value  of  the  archaeological
     resource.  The  magnitude of  this  damage  depends  on  the
     uniqueness  of  the  affected  site, the  original quality of
     information available at  the site, the nature of the impacts,
     and the willingness of the scientific community to pay for the
     lost information.    If  the  site is  unique  and  substitute
     sources of similar information do not exist, the value of the
     damage may be large.

2.   Loss of value as tourist  and educational attractions.  Unique
     or  spectacular archaeological sites  have  value  as tourist
     attractions.    All significant  archaeological  sites  have
     educational value  as  the focus of  field trips and published
     descriptions.   Archaeological information and  artifacts have
     value  for  museum  interpretation  and  display.  Oil impacts
     could substantially reduce these values.
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Intrinsic Value

1.   Impacts on  the religious,  cultural  or symbolic  values for
     native groups.

2.   Loss  of   intrinsic  value  for  the   general,   non-native
     population.


                              BUDGET

This study has not yet begun due to the delay in receiving results
from the archaeological science study.  At present, no additional
funds have been requested  for this study.   When results from the
archaeological resources  science study are  received,  this study
will be continued  and  funded with the amount needed to determine
the  extent  of  both  the  lost  use  and  intrinsic  values  of
archaeological resources.
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ECONOMIC STUDY NUMBER 10

Study Title:  Petroleum Products Price Impacts


                           INTRODUCTION

Retail prices for gasoline on the West Coast of the United States
increased immediately after the  EVOS.   This increase is observed
both relative to earlier periods  in 1989 and relative to prices in
other parts of the country immediately  after  the  spill.   Similar
increases in other petroleum products may also have occurred.


                           OBJECTIVES

Estimate economic damages to consumers of petroleum products.


                             METHODS

This study will conduct  a statistical analysis of the relationship
between the EVOS  and the observed petroleum market price increases.
If it  appears that a  connection between the  two events  can be
shown,  the  damage to   consumers of  petroleum products  will be
estimated.  Investigators will use existing data and models as well
as improved data and methods they develop to value the injury.


                             BUDGET

Total:                      $ 271.3
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PART IV:  OIL SPILL PUBLIC INFORMATION SUPPORT

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OIL SPILL PUBLIC INFORMATION SUPPORT

Lead Agency:   DOJ, DOA, DOI, NOAA


                           INTRODUCTION

The  Federal trustee  agencies are  committed to  making  as  much
information about the EVOS available to the public as possible.


                            OBJECTIVES

A.   Provide a central  facility  for collecting information about
     oil spills in general and the EVOS in particular.

B.   Gather scientific  data  from each  of the government agencies
     involved in  the spill  response or natural  resource damage
     assessment.

C.   Answer Freedom  of  Information Act requests  from the public
     about the EVOS.


                             METHODS

The  Oil  Spill Public  Information  Center  (OSPIC)  is  located in
Anchorage, Alaska,  and was  opened on September 27,  1990.  The OPSIC
serves the  public  by providing  access to information  about oil
spills in general  and the Exxon Valdez oil  spill in particular.
The  current  collection  includes  technical  reports,  newspaper
clippings,  maps,  slides,  photographs,  books,  periodicals,  audio
recordings, and video tapes.   The OSPIC has received requests from
corporate entities,  students, college faculty, the  legal community,
and  members of  the public.   The  OPSIC will  begin  to  catalog
scientific data from the EVOS during 1991.  OSPIC staff will also
continue to process documents collected in response to Freedom of
Information  Act  (FOIA)  requests  for inclusion in the  OSPIC
collection.


                              BUDGET

The trustee agencies will  reimburse the  Department of Justice in
equal shares for the operation'of the OSPIC and according to agency
activity for FOIA processing.

Department of Agriculture                           $  614.0
Department of Interior                                1,739.0
National Oceanic and Atmospheric Administration        599.0

Total                                               $2,952.0

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FART V:  RESTORATION PLANNING

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                       RESTORATION PLANNING

                            OBJECTIVES

The  goal  of the  restoration  planning  effort is  to  identify
appropriate measures that can be taken to restore natural resources
injured by the EVOS.  Specific objectives are:

A.   Identify or develop technically feasible restoration options
     for natural resources  and services  potentially injured by the
     EVOS;

B.   Determine the nature and pace of natural recovery of injured
     resources,  and identify where direct restoration measures may
     be appropriate;

C.   Incorporate an approach to restoration that  where appropriate,
     focuses  on  recovery  of  ecosystems  rather  than  on  the
     individual components of those systems;

D.   Identify  costs  associated  with  implementing  restoration
     activities, in support of the overall natural resource damage
     assessment process; and

E.   Encourage,   provide   for,   and  be  responsive   to  public
     participation  and review  during  the  restoration  planning
     process.
                            DEFINITION

For any  injury, there  are three  types  of possible  restoration
activities:

1.   direct restoration refers to measures in addition to response
     actions, usually taken on  site, to  directly rehabilitate an
     injured, lost, or destroyed resource;

2.   replacement  refers  to  substituting  one  resource  for  an
     injured, lost, or destroyed resource of  the same or similar
     type; and

3.   acquisition of ecfuivalent resources means to compensate for an
     injury to  a resource by substituting another  resource that
     provides the  same  or substantially similar  services  as the
     resource injured, lost, or destroyed.

     Determining the adequacy  of natural recovery is fundamental to
     the choice of a restoration  activity.    In some  cases the
     Trustees maydetermine  that it is most appropriate to allow
     natural recovery to proceed without further intervention.
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                   1990 RESTORATION ACTIVITIES

The Trustee agencies  and  EPA initiated several small-scale field
studies  to  evaluate  the  feasibility of  restoration techniques.
Results  from  these  studies will help to  determine  the costs and
effectiveness  of  full-scale   restoration   projects.     Several
technical  support   studies  were  also  initiated  to  provide
information  needed  to  evaluate  or   carry  out  some  potential
restoration activities.  These studies were described in the 1990
State/Federal Natural Resources Damage Assessment and Restoration
Plan for the Exxon Valdez Oil Spill. August 1990 (available at the
OSPIC) and preliminary results are summarized below.


               1990 RESTORATION FEASIBILITY STUDIES

1.   Reestablishment of Fucus in Rocky Intertidal Ecosystems

     Lead Agency:  EPA

Early observations indicated that Fucus, a marine plant  (rockweed)
found on rocky shorelines  in the intertidal zone throughout the oil
spill area, was  extensively damaged by both the  spilled oil and
cleanup  efforts.    If  the natural  recovery  of  Fucus  could  be
significantly  accelerated  or  enhanced,  it  would benefit  the
recovery of associated flora and fauna  on  intertidal rocky shores.

Specific objectives of  this  study were to identify the causes of
variation in Fucus recovery at and near Herring Bay, Knight Island
in PWS; to document the effects of alternative  cleaning methods on
Fucus; and to  test the feasibility of enhancing  the reestablishment
of Fucus.  Although results are preliminary it appears that Fucus
recovers most  slowly  at intensively cleaned sites  and almost no
recovery occurs where tar cover persists.

2.   Reestablishment  of   Critical  Fauna  in  Rocky  Intertidal
     Ecosystems

     Lead Agency:  USFS

This feasibility study was designed to  compare  the rates of faunal
recovery in rocky intertidal communities, and to demonstrate the
feasibility  of  restoration of  these  communities  by  enhancing
recolonization rates for such key species  as  limpets and starfish.
Recolonization rates  for these  organisms and for  the rockweed,
Fucus, may  limit the  natural rates of  recovery for  the entire
community.  Parameters  examined included  the presence or absence
ofcommon  intertidal  species on  impacted  and reference  sites,
population dynamics  of several species of  invertebrates,  larval
settlement on  oiled  versus unoiled surfaces,  and differences in
algal grazing by  limpets between oiled and reference sites.  One of
the preliminary results indicates that heavy predation of several

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species of transplanted invertebrates was probably due to the lack
of cover usually provided by Fucus.

3.   Identification  of  Potential  Sites  for  Stabilization  and
     Restoration with Beach Wildrye

     Lead Agency:  DNR

This study  was designed to  identify  sites  with injury  to beach
wildrye grass and to recommend restoration measures.  Beach wildrye
grass is  important in the prevention  of erosion in  the coastal
environment  and is  a key  component  of  supratidal habitats  in
locations throughout the oil  spill area.  Erosion  resulting from
loss  of  beach  wildrye can  lead  to  the  destabilization  and
degradation of wildlife habitats and of cultural and recreational
sites.   Results from survey work conducted in 1990 in PWS indicate
injury to several beach wildrye communities.

4.   Identification of Upland Habitats Used by Wildlife Affected by
     the Oil Spill

     Lead Agency:  FWS, ADF&G

A diversity of birds, mammals, and other animals were killed by the
spill or injured by contamination of prey and  habitats.   Many of
these species are dependent on aquatic or intertidal habitats for
activities such as feeding and  resting,  buy many also use upland
habitats.  Protection of upland habitats from further degradation
may reduce  the effects of the  oil  on injured  fish and  wildlife
populations, and thereby speed their recovery.  This study focused
specifically on marbled murrelets and harlequin ducks, two species
known to have  been affected  by  the spill and known to use upland
habitats.

Based  on surveys  of  140  streams,  preliminary results  of  the
harlequin duck study indicate that this species  nests along  larger-
than-average anadromous fish streams,  with moderate gradients and
clear  waters.    Preliminary  results  on murrelets suggest  that
murrelets use north facing slopes, and inland areas  at the heads of
bays.  Open bog meadows, especially at the heads of bays,  appear to
be used as flight corridors to upper wooded areas.

5.   Land Status, Uses,  and Management Plans  in  Relation to Natural
     Resources and Services

     Lead Agency:  DNR

The objective of this study is to locate,  categorize, evaluate, and
determine the  availability of maps,  management  plans,  and other
resource documents relevant to restoration planning  throughout the
oil spill region.   Resource materials  identified  will  assist in
planning  for implementing site-specific restoration activities,

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including direct restoration, replacement, and the acquisition of
equivalent resources.

To date, a variety  of  documents,  maps,  and management plans have
been identified and are being evaluated; other resource materials
are being located.  This preliminary project will be completed in
Spring 1991.  A second phase is under consideration.

1990 Technical Support Projects

1.   Peer Reviewer Process for Restoration Feasibility Studies

     Lead Agencies:  ADF&G, DEC, DNR, DOI, DOA, NOAA, EPA

This  project  provided  funds  to  ensure  that  scientists  with
expertise on natural resource restoration were available to provide
peer  review  of  restoration  feasibility  projects  and  other
restoration planning studies and activities.

2.   Assessment of Beach Segment Survey Data

     Lead Agency:  DNR

The objective  of this project  is to review  and  summarize beach
survey information (obtained through oil spill response activities)
to  assist   in  planning  for  and  implementing  site-specific
restoration  activities,  particularly  in  the  area  of  direct
restoration.  This study was  initiated late in 1990 and continues.

A master database is being created from that portion of the beach
surveys relevant  to restoration.     The primary sources  of this
information  are  DNR  and  DEC.    Data  from  local   and  regional
governments  as well  as  non-governmental  sources  will also  be
reviewed and integrated  into the system  as appropriate.   This
preliminary project will be completed in Spring 1991.

3.   Development of Potential Feasibility Studies for 1991

     Lead Agencies:  ADF&G, EPA

This project provided  for the  orderly  development  of additional
feasibility studies including: a)  monitoring  "natural" recoveries;
b)   pink  salmon  stock   identification;   c)   herring   stock
identification/spawning  site  inventory; d)  artificial  reefs for
fish and shellfish;  e) alternative recreation sites and facilities;
f) historic  sites and artifacts;  and g)  availability  of forage
fish.     Feasibility   study   proposals   are  currently   under
consideration including the above topics.
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               1991 RESTORATION PLANNING ACTIVITIES

The fundamental purpose  of restoration planning  is  to identify,
evaluate, and then recommend potential restoration implementation
activities, in consultation with technical experts and the public.

The NRDA studies  and  other sources  (e.g.,  Shoreline  Assessment
Program, and other agency surveys not connected with the oil spill)
provide information on  species, habitats,  and ecosystems in need of
restoration.    In   1991,   as  damage  assessment   results  are
synthesized,   the   RPWG   will   consult   with   the   principal
investigators, agency experts, and outside peer reviewers to review
the nature  and extent  of  oil spill injuries in  relation  to the
biology and ecology of the injured resources.  A key goal in this
process  will  be to identify life history  requirements,  limiting
factors, and environmental  processes that are especially sensitive
or  that  may  be  enhanced.   In  turn,  this will   lead  to  the
identification of potential restoration activities.

Once  potential restoration  implementation  activities have been
identified,  they  must  be evaluated   in   terms  of  technical
feasibility, environmental  benefit,  cost, and other  factors.   In
1991,  the RPWG will  continue to evaluate the restoration options
identified thus far  (e.g.,  those  presented  in RPWG's Restoration
Planning Following the  Exxon Valdez Oil Spill;  August  1990 Progress
Report), as well as new options that are suggested through public
and technical consultations.

While  some  potential  restoration implementation activities  are
readily evaluated, others require more detailed review and study.
In some  cases,  the  RPWG will recommend that restoration science
studies   (feasibility,  monitoring,   or  technical   support)   be
conducted to test the efficacy of particular options or to gather
basic  information necessary to evaluate or  implement an  option
(e.g.,  biological or  resource  assessment   data).    Several such
studies were carried out in 1990.  Subject to additional technical
review and availability of  funds,  some restoration science studies
and implementation projects are being considered in 1991.  If these
studies or projects are carried forward they will  be  outlined in a
Federal Register notice later this spring.  Additional information
on the Trustees' plan to implement restoration projects  in 1991 was
provided in the March 1, 1991,  Federal Register.   (56 FR 8898) .

The RPWG also expects to further evaluate restoration approaches.
For example, the RPWG will  review  different management systems for
protecting  marine  habitats  (e.g.,  National  Marine  Sanctuary
Program, Alaska Marine Parks).   Another example would be to carry
out   economic  and    environmental  analyses    of   restoration
alternatives.
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As information about injuries becomes available, and as potential
restoration   actions  are   evaluated,    further   implementation
activities may be recommended.

Literature Review

The  scientific  literature and information from other  oil spills
will provide background information that is helpful in restoration
planning.   In  1991,  the RPWG  expects  to  synthesize  previously
identified literature on restoration (see Appendix B, August 1990
Progress Report).  The RPWG will also complete previously initiated
syntheses  of  literature  on species   and  ecosystem  recoveries
following natural and human-induced environmental disturbances.

Monitoring

Information  on  the  adequacy of natural  recovery is  central  to
determining whether to implement restoration activities  or to allow
injured resources to recover on  their own.  The literature reviews
described  above will  provide  background  information for  such
considerations,   while  damage  assessment studies  will  provide
current data on the  status  of resources injured by the EVOS.   In
1991 the RPWG expects to recommend several monitoring studies to be
carried  out  in  the  field in 1991  and  to develop  protocols  for
evaluating the effectiveness of any restoration projects that are
implemented.  The  RPWG also will  continue efforts  to develop a
comprehensive plan for long-term ecological monitoring that could
be implemented in the oil spill environment following resolution of
damage claims.

Public Participation

In 1990, the RPWG  emphasized broad  scoping activities to invite
suggestions from the public about potential restoration activities
and priorities.  Public participation will continue to be important
in 1991, with increased emphasis on evaluating and determining the
importance of restoration  alternatives.  The RPWG is interested in,
and  available   for,  meetings  with  individuals or  constituency
groups.  There also will be consideration of additional activities,
such  as  publications  and  workshops   in 1991.    Requests  and
suggestions from the public  are invited.

Scientific Review

Technical review is  essential to the scientific integrity of the
restoration  planning process.   As needed, the RPWG  draws  upon
experts from academic institutions,  public  agencies,  and private
organizations (e.g.,  consulting firms, non-profit organizations) as
sources  of  advice   and  criticism  in  planning feasibility  and
technical support studies, and in evaluating and recommending
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restoration activities.  In 1991, the RPWG will continue to place
emphasis on  scientific review,  including participation  by peer
reviewers.
                          BIBLIOGRAPHY

Trustee Council.  1990.   1990 State/Federal Natural Resource Damage
     Assessment  and  Restoration Plan  for the  Exxon Valdez  Oil
     Spill; August, 1990.  360pp plus appendices.

Restoration  Planning Work  Group.   1990.    Restoration  Planning
     Following the Exxon Valdez Oil Spill; August  1990 Progress
     Report.  80 pp.


                             BUDGET

The following restoration planning budget does  not include the 1991
costs of any potential restoration implementation projects.

Salaries:                               $835.0
Travel:                                  250.0
Supplies:                                 20.0
Equipment/Office:                         75.0
Contractual Services:
     Literature Review                   125.0
     Scientific Review                   100.0
     Public Participation                 30.0
     Restoration Options Analysis        200.0
     Report Publications                  25.0
Restoration Science Studies:            3.875.0

Total Planning Activities Budget:     $5,485.0
                               282

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PART VI:  BUDGET

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Budget Summary for the Exxon Valdez Oil
     Budgeted  costs  for projects  from 3-
Spill Damage Assessment - 1991
1-91 through 2-29-92.
STUDY
NO.
Marine
2
4
5
6

STUDY TITLE
Mammals
Killer Whale
Sea Lion
Harbor Seal
Sea Otter Injury

LEAD AGENCY

NOAA
ADF&G
ADF&G
DOI
subtotal
BUDGET

$186,000
24,000*
94,200
810,800
$1,115,000
Terrestrial Mammals
3
4
6

Birds
1
2
3
4
11

River Otter & Mink
Brown Bear
Mink Reproduction


Beached Bird Survey
Census/Seasonal
Distribution
Seabird Colony Surveys
Bald Eagles
Sea Ducks

ADF&G
ADF&G
ADF&G
subtotal

DOI
DOI
DOI
DOI
DOI
subtotal
$377,300
76,000
8,500*
$461,800

$313,000
220,000
530,000
255,000
178,900
$1,496,900
                                    283

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Budget Summary for the Exxon Valdez oil Spill Damage Assessment -  1991
                                (continued)
     Budgeted costs  for projects from 3-1-91 through 2-29-92.
STUDY
NO.
STUDY TITLE
LEAD AGENCY
BUDGET
Fish/Shellfish
1
2
3
4
5
7
Salmon Spawning Area Injury
Eggs /Pr e-emergent Fry Sampling
Coded-wire Tagging
Early Marine Salmon Injury
Dolly Varden Injury
Salmon Spawning Area
ADF&G
ADF&G
ADF&G
ADF&G
NOAA
ADF&G
ADF&G
$288,000
259,000
1,075,000
136,400
172,000
325,100
15,000*
   8



   11

   13

   15

   17

   18

   24

   27

   28

   30
Injury, Outside PWS

Egg & Pre-emergent Fry
Sampling, Outside PWS

Herring Injury

Clam Injury

Injury to Shrimp

Injury to Rockfish

Trawl Assessment

Injury to Demersal Fish

Sockeye Salmon Overescapement

Run Reconstruction

Database Management
ADF&G



ADF&G

ADF&G

ADF&G

ADF&G

NOAA

NOAA

ADF&G

ADF&G

ADF&G

subtotal
       15,000*


      558,000

      147,000

moved to Subtidal

moved to Subtidal

       40,000*

moved to Subtidal

      334,300

      175,100

      175,800

   $3,715,700
                                    284

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Budget Summary for the Exxon Valdez Oil Spill Damage Assessment - 1991
                                (continued)
     Budgeted  costs  for projects from 3-1-91  through 2-29-92.
STUDY
NO. STUDY TITLE
LEAD AGENCY BUDGET
Coastal Habitat
1A
IB
Intertidal Studies
Intertidal Studies
Air/Water
2a
2b
3
6
Injury to Subtidal
Deep Water Benthos
Hydrocarbon in Water
Oil Fate and Toxicity
USFS
NOAA
subtotal
DEC
NOAA
ADF&G
DEC
NOAA
NOAA
$5,100,000
68,000
$5,168,000
moved to Subtidal
moved to Subtidal
moved to Subtidal
moved to Subtidal
moved to Subtidal
moved to Subtidal
Subtidal
1
2
3
4
5
6
7
Hydrocarbon Exposure, Microbial
and Meiofaunal Community Effects
(A/W 2a)
Injury to Benthic Communities:
Bio-availablity and transport
of hydrocarbons (A/W 3)
Sediment Toxicity Bioassays (A/W 6)
Injury to Shrimp (F/S 15)
Injury to Rockfish (F/S 17)
Injury to Demersal Fish (F/S 24)
DEC
NOAA
ADF&G
DEC
NOAA
NOAA
ADF&G
ADF&G
ADF&G
NOAA
$139,800
295,000
592,500
196,200
150,000
125,000
50,000
120,000
80,000
235,000
                                                subtotal
$1,983,500
                                    285

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Budget Summary for the Exxon Valdez oil spill Damage Assessment - 1991
                                (continued)
     Budgeted  costs  for projects from 3-1-91  through  2-29-92.

   STUDY
    NO.	STUDY TITLE	LEAD AGENCY	BUDGET	

   Technical Services

   1        Hydrocarbon Analysis               DOI             $550,000
                                               NOAA          2,000,000

   3        Mapping                           DOI             300,000
                                               ADNR            656,300

                                               subtotal     $3,506,300

   Archaeology

   1        Archaeological                     ADNR            $688,600
                                               USFS            103,000

                                               subtotal        $791,600

            SUBTOTAL  FOR SCIENCE PROJECTS                   $18,238,800

   Peer Reviewers/Chief Scientist

            Department of Agriculture                          $772,000
            Department of Interior                             772,000
            National  Oceanic and Atmospheric Administration   772,000

            SUBTOTAL  FOR PEER REVIEWERS/CHIEF SCIENTIST     $2,316,000

   Economics

   1        Commercial Fisheries Losses        FEDERAL         $265,500
   5        Recreation Uses Damage             FEDERAL         390,400
   6        Subsistence Losses                 FEDERAL         532,100
   7        Intrinsic Value Loss               FEDERAL       1,964,600
   8        Research  Program Damage            FEDERAL         104,900
   10       Petroleum Products Price           FEDERAL         271,300

            SUBTOTAL  FOR ECONOMICS                          $3,528,800

   Restoration Planning

            State of  Alaska                                 $2,968,000
            Environmental Protection Agency                  1,267,000
            Department of Interior                             300,000
            Department of Agriculture                          525,000
            National  Oceanic & Atmospheric Administration     425,000

            SUBTOTAL  FOR RESTORATION                        $5,485,000**

                                    286

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Budget Summary for the Exxon Valdez oil  Spill Damage Assessment - 1991
                                (continued)
    Budgeted  costs for projects from 3-1-91 through 2-29-92.

   STUDY
   NO.	STUDY TITLE	LEAD AGENCY	BUDGET

   Oil Spill Public Information Support

       Department of Agriculture                              $614,000
       Department of Interior                                1,739,000
       National Oceanic and Atmospheric Administration         599,000

       SUBTOTAL FOR OIL SPILL PUBLIC INFORMATION SUPPORT    $2,952/000

   Overhead

       State  of Alaska                                      $1,037,200
       Department of Agriculture                               600,000
       Department of Interior                                  300,000
       National Oceanic and Atmospheric Administration         900,000
       Environmental Protection Agency                         200,000

       SUBTOTAL FOR OVERHEAD                                $3,037,200
                                     GRAND TOTAL       $35,557,800
  * These studies are being funded for the completion of data analysis and
  final report preparation.

  **   Restoration implementation projects may be  conducted  this summer
  depending on resource availability.  (See FR 88, 98, March 1, 1991.)
                                    287

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               BUDGET SUMMARY FOR THE EXXON VALDEZ OIL SPILL BY AGENCY
State of Alaska                                                 $10,612,300



Department of Agriculture                                         7,714,000



Department of Interior                                            6,268,700



National Oceanic and Atmospheric Administration                    5,967,000



Environmental Protection Agency                                    1,467,000



All Federal Agencies (Economics)                                   3,528,800
                         GRAND TOTAL                         $35,557,800
                                        288

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APPENDICES A, B AND C

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                            APPENDIX A

               STATE/FEDERAL DAMAGE ASSESSMENT PLAN

                       ANALYTICAL CHEMISTRY

                QUALITY ASSURANCE/QUALITY CONTROL


This document  describes the Quality Assurance for the analytical
chemistry portions of the Exxon Valdez Damage Assessment Process.
It  is  to be  used in  conjunction with the  Analytical Chemistry
Quality Assurance Programs of the Trustee Agencies.  It describes
only those  minimum  requirements  necessary  to validate  the data
generated by analytical chemistry laboratories.  Quality assurance
requirements for other types of measurements are not addressed.

For  instructions  in meeting  the requirements described  in this
document, please  consult  "Collection and Handling  of Samples",
which was prepared  by the Analytical Chemistry Group  for use in
training field personnel or the following Agency representatives:

Carol-Ann Manen, National Oceanic and Atmospheric Administration

Everett Robinson-Wilson, U.S. Fish and Wildlife Service


                        TABLE OF CONTENTS

1.   QUALITY ASSURANCE FOR ANALYTICAL CHEMISTRY

     1.1  Study-Specific QA Plans
     1.2  Technical  System Audits
     1.3  Standards  and Quality Control Materials
     1.4  Analytical Performance Evaluations
     1.5  Data Reporting and Deliverables

2.   MINIMUM REQUIREMENTS:  SAMPLING AND SAMPLING EQUIPMENT

     2.1  Sample Identification and Labelling
     2.2  Sample Field Chain-of-Custody

3.   MINIMUM REQUIREMENTS:  ANALYSIS

4.   MINIMUM REQUIREMENTS:  REPORTING AND DATA DELIVERABLES
                               A-l

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1.   Quality Assurance for Analytical Chemistry

Each Trustee agency through their individual standard documented QA
programs and guidances shall ensure that all  data generated by or
for that  agency and their  contractors,  in  support of  the Exxon
Valdez Damage Assessment,  are of known, defensible, and verifiable
quality.

These documented  QA programs and  guidances  include but  are  not
limited to:

     NOAA National Status and Trends Program, Mussel Watch Phase
          4 Work/QA Project Plan
     Quality Assurance of Chemical Analyses  Performed Under
          Contract With the USFWS
     EPA SW-846, Chpt. 1, QA/QC Requirements
     EPA Guidelines and Specification for Preparing Quality
          Assurance Project Plans, QAMS-005
     EPA Handbook for Sampling and Sample Preservation of Water
          and  Wastewater

The principal  investigators for  Technical  Services Study  1,  in
consultation  with  expert  scientists  developed  and  oversee  a
centralized program to demonstrate the quality and comparability of
the chemical data obtained by the Trustee agencies.

The major components of this centralized QA program will be:

1.   Development of study-specific analytical chemistry QA plans.

2.   Technical on-site system audits of field and laboratory data
     collection activities.

3.   Development   and   provision   of   appropriate   instrument
     calibration standards and control materials.

4.   Laboratory performance evaluations by means of intercomparison
     exercises.

5.   Review of data deliverables and all supportive documentation
     to evaluate data quality.

1.1  Study-Specific Quality Assurance Plans

Prior to the initiation of each study, the principal investigator
must prepare and submit a study-specific analytical chemistry QAP
to  Technical Services  1  principal  investigators  and scientific
experts for review and concurrence.  This plan shall specify each
study's goals, sampling procedures, analytical procedures,  and all
quality control measures  and  acceptance criteria associated with
those procedures.


                               A-2

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The QAP must be study-specific, however any documented QA guidance
and/or appropriate Standard Operating Procedures  (SOP's)  used by
the Trustee  agencies  may form the  basis of individual  study QA
plans.

A Quality Assurance Plan must address the following:

     *  Title Page -  Includes the signatures of  the individuals
        responsible  for the  project  and Technical Services  1
        concurrence.

     *  Project  Description   and  Sampling Objectives  -  Briefly
        describes the what,  where, and why of the project.

     *  Data Needs - Describes what elements,  compounds, classes of
        compounds,  and/or physical  data are  required.    Must
        describe  the  desired detection limits,  precision  and
        accuracy of the data for the study.

     *  Sampling  and  Labelling   Procedures  -  Describes  sample
        collection,  including field QC and preservation.  Estimates
        the number and kind of samples to be collected.   Minimum
        requirements  for  sample  collection  are  described  in
        Section 2.

     *  Chain   of  Custody   -  Describes  Chain-of-Custody  and
        documentation  procedures.    Minimum   requirements  are
        described in Section 2.

     *  Analytical Procedures - References or describes in detail
        proposed method(s).

     *  Internal Quality Control  - Describes  type and frequency of
        internal  quality  control.     Minimum  requirements  are
        described in Section 3.

     *  Calibration  Procedures   and  Frequency  -  Describes  the
        methods and frequency for  calibrating field and laboratory
        instruments.   These must be specified in SOP's.

     *  Data Verification - Describes the data verification in SOP
        form and  includes;  (1)  the methods  used  to identify and
        treat outliers, and  (2) the  data flow  from generation of
        raw data through storage of verified results.

     *  Data Deliverables - Specifies reporting needs additional to
        the minimum requirements described in Section 4.

     *  Technical System and Performance Audits - Specifies field
        or  intra-laboratory   audits  planned  by the responsible
        agency.
                               A-3

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1.2  Technical System Audits

On-site system audits may be performed without prior notification
by  the  Technical   Services   l   principal   investigators  after
consultation with the responsible agency.


1.3  Standards and Quality Control Materials

The National  Institute of  Standards  and  Technology  (NIST)  will
develop  and provide  appropriate standards  and quality  control
materials.
1.4  Analytical Performance Evaluations

Prior to the initiation of work, each analytical laboratory will be
required to demonstrate its capability.  This will be accomplished
by providing  laboratory  documentation on the performance  of the
proposed methods  and through  the  analysis  of an  accuracy based
material.  The results of this analysis must be within +/- 15% of
the value of each analyte or measurement parameter.

Any changes  in analytical methodology from that proposed  in the
original QA  plan  shall be validated under  agency  procedures and
documented to the  Technical Services 1 principal investigators and
expert scientists.

A series of  three intercomparison  exercises,  utilizing the blind
analysis  of  gravimetrically  prepared  materials,  extracts  of
environmental matrices (tissue, sediment and water)  or the matrices
themselves, will  be  conducted annually.  Participation  in these
exercises is mandatory.   Materials  will be  prepared by,  and data
returned to the  NIST  for statistical  analysis.   The NIST will
report  to  the Technical  Services 1  principal  investigators.
Unacceptable  performance will result  in the  discarding  of the
associated data.

The Technical  Services 1 principal  investigators  will review and
provide written reports on the results of intercomparison studies
to the Management Team.


1.5  Data Reporting and Deliverables

Data deliverables will be  reviewed by the generating  agency to
verify the quality and usability of the data.  A QC report on each
data set will  be  provided to  the Technical Services  1 principal
investigators for review.


All data  and associated  documentation  will be held  in  a secure

                               A-4

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place under chain-of-custody procedures until the Trustees indicate
otherwise.
2.   Minimum Requirements;  Sampling and Sampling Equipment

Sample  collection   activities  must  be   described   in  SOP's.
References to existing documents are acceptable.

The method of collection should not alter the samples.

Sample collection and storage devices shall not alter the sample.

Samples  shall  be  held  in  a secure  place  under  appropriate
conditions and under chain-of-custody until the Trustees indicate
otherwise.


2.1  Sampling Identification and Labelling

An SOP will be in place for each study which describes procedures
for the unique  identification of  each  sample.   A  sample  tag or
label will  be attached  to the sample  container.   A waterproof
(indelible)  marker must be used on the tag or label.  Included on
the tag are the sample identification number, the location of the
collection  site,  the  date of  collection  and  signature  of  the
collector.

The information above  will also be recorded in  a  field notebook
along with  other  pertinent information about the  collection and
signed by the collecting scientist.


2.2  Field Chain-of-Custody

The field sampler will be personally responsible for the care and
custody of  the  samples  collected  until they are  transferred to
another responsible party.

Samples will be accompanied by a chain-of-custody record or field
sample  data  record.    When  samples  are  transferred   from  one
individual's custody to  another's,  the individuals relinquishing
and receiving will sign, date and note the time on the record.

Shipping containers will be custody-sealed for shipment.  Whenever
samples are split,  a  separate chain-of-custody record will be
prepared for those  samples and marked to  indicate  with whom the
samples are being split.


Samples shall  be  maintained   in  a manner that preserves their
chemical integrity from collection through final analysis.

                               A-5

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Sample shipper will arrange for sample receipt.

After analysis, any remaining  sample  and  all  sample tags,  labels
and containers shall be held under chain-of-custody procedure until
the Trustees indicate otherwise.
3.   Minimum Requirements;  Analysis

The applicable  methodology shall be  referenced or  described in
detail in the SOP's for each measurement parameter.

Method  limits  of  detection shall  be calculated  by matrix  and
analyte.

Control of the analytical method in terms of accuracy and precision
shall be demonstrated.

Calibration shall be verified at the  end of each analysis sequence.

Samples shall be quantified within the demonstrated linear working
range for each analyte.

Standard curves  shall  be established with at least 3 points besides
0.

Field blanks, procedural  blanks,  reference materials,  replicates
and analyte recovery samples shall be run at a minimum frequency of
5 percent each per sample matrix batch.

A minimum list of the  petroleum hydrocarbon compounds which are to
be  considered  for  identification and  quantification  in  water,
tissue and sediment include the volatiles, i.e., benzene, toluene,
xylene  and the  polynuclear aromatic  and aliphatic  hydrocarbons
listed below:

     Naphthalene                        n-dodecane
     2-Methylnaphthalene                n-tridecane
     1-Methylnaphthalene                n-tetradecane
     Biphenyl                           n-pentadecane
     2,6-Dimethylnaphthalene            n-hexadecane
     Acenaphthylene                     n-heptadecane
     Acenaphthene                       pristane
     2,3,5-Trimethylnaphthalene         n-octadecane
     Fluorene                           phytane
     Phenanthrene                       n-nonadecane
     Anthracene                         n-eicosane
     1-Methylphenanthrene
     Fluoranthene
     Pyrene
     Benz(a)anthracene
     Chrysene

                               A-6

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     Benzo(b)fluoranthene
     Benzo(k)fluoranthene
     Benzo(a)pyrene                     Benzo(e)pyrene
     Indeno(1,2,3-c,d)pyrene            Perylene
     Dibenz(a,h)anthracene
     Benzo(g,h,ijperylene
4.   Minimum Requirements;  Reporting and Data Deliverables

Measurement  results,   including  negative  results,  as if  three
figures were significant shall be reported.

Results of quality control samples analyzed in conjunction with the
study samples shall be reported.

Documentation demonstrating  analytical control of precision and
accuracy on an analyte  and matrix specific basis shall be reported.
                               A-7

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                            APPENDIX B

                   EVOS DAMAGE ASSESSMENT PLAN
                    HISTOPATHOLOGY GUIDELINES

Histopathology is an important tool  used in determining mechanisms
of  death  and sublethal effects  caused by infectious  agents and
toxic substances.   A definitive diagnosis often  does  not result
from histological examination,  but can give strong support to other
positive  measurements.    Tissues deteriorate  (autolyze)  rapidly
after an animal dies; therefore,  to  be of value, any samples taken
for histological evaluation as  part of the damage assessment of the
EVOS  shall  be collected,  preserved,  and processed  under strict
guidelines.

Sample Collection and Preservation Protocols

Standard protocols for necropsy and  preservation of tissue samples
for  histopathology  shall  be used  throughout  the NRDA studies.
Different protocols have been designed to accommodate the different
groups of animals to be  encountered in the  assessment studies.
Necropsy  procedures  have  been established and  provided to study
managers  under  separate cover for a  variety  of different animal
groups including  finfish,  bivalve mollusks,  brachyuran and crab-
like anomurans (i.e., king crabs), shrimp, marine and terrestrial
mammals, and migratory and nonmigratory waterfowl.

Paired sampling of animals from oiled versus unoiled sites will be
done for comparative purposes.  Histopathological sampling should
be  done  during  any observed  acute  episodes of  mortality  or
morbidity to determine  the  cause of death or abnormality.  These
types of samples are the most valuable in assessing acute toxicity
affects and will be the most  likely samples collected  for birds and
mammals  due to  their  high visibility  in  the  impacted  areas.
Because of the  low visibility of fish and shellfish, many histology
samples will consist of random collections  in impacted and control
areas  with  little  prior  obvious   indication  of  morbidity  or
mortality.

Any histological  processing  of samples collected from apparently
normal  shellfish will  be  performed after,  results  of parallel
hydrocarbon sampling are known; i.e.,  positive hydrocarbon results
may  merit further  histopathology  studies.   This  would  not  be
advisable for fish and other higher  animals that possess an active
mixed  function  oxidase   (MFO)  liver enzyme  system  which  could
metabolize  hydrocarbons  to  other  compounds providing negative
hydrocarbon   results,   while   potentially   still   exhibiting
toxicological lesions.  Analyses of  enzyme  function may show an
activated  MFO   system  in   exposed  fish  and  higher  animals.
Consequently, histology and  hydrocarbon samples, as well as other
appropriate samples, such  as  liver and bile, will be taken  from the

                               B-l

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same animal when possible  for  analyses  of  metabolites and enzyme
function.   If certain fish and  shellfish  are too few  or small,
subsampling other animals from  the  same  site at the same time will
be necessary.

Processing and Interpretation Protocols

Histopathology  assessment  of  birds  and  mammals will  be  done
primarily  on  tissues  from clinically affected  animals  using
established criteria of cellular  degenerative and necrotic changes
recognized by a board certified veterinary pathologist.

Histopathological analysis  of  finfish and  shellfish  tissues will
include the criteria  above  as  well as  indices established in the
Amoco Cadiz oil spill studies  (Haensly et al. 1982; Berthou et al.
1987)  to   allow  some  quantification  of   potentially  subtle
degenerative changes  in  tissue histology  of  otherwise clinically
normal animals.  Briefly, these indices  include mean concentration
of mucus cells per mm2 of gill  lamellae  (fish); mean concentration
of mucus  cells per  mm  of  epidermis in  10  fields  (fish) ;  mean
concentration  of  macrophage  centers  per  mm  of  liver;  mean
concentration   of   hepatocellular   vacuolation   due  to   fatty
degeneration  (fish);  a  mean  and  total   tissue  necrosis  index
(invertebrates); histological  gonadal  index  (invertebrates); and
differences in prevalences  and intensities of incidental lesions
caused by infectious agents (fish and invertebrates).

Quality  Assurance   in  Field   Collection  of  Samples   and  in
Interpretation of Results

Field Collection:

Veterinary personnel trained in sample taking will be utilized for
onsite necropsies of birds and  mammals in order to ensure adequate
quality control and standardized  sample  collection.  The same high
standards will  be attainable  in fish  and invertebrates  in that
sample collection will be done by  trained finfish  and shellfish
biologists.   A fish pathologist and technician are  available to
train field personnel and  assist in necropsy and  preservation of
finfish and shellfish samples at collection sites.

Finfish and shellfish samples can be coordinated through an ADF&G
fish  pathologist,   Fisheries  Rehabilitation,   Enhancement  and
Development Division.

Interpretation of Results:

Quality control  of all  processed  work will  require independent
blind reading of  subsampled  histology slides  by two  different
laboratories.    Tissues  with  known lesions  will  be  included
                               B-2

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periodically  in  groups of tissue  samples for blind  reading and
determination of competency in interpretation.

Chain-of-Custody Guidelines

Due to the evidentiary nature of sample collecting investigations,
the possession  of samples will  be traceable  from the  time the
samples are  collected until they  are introduced as  evidence in
legal proceedings.   To maintain and  document  sample possession,
chain-of-custody procedures will be followed.

The field sampler will be personally responsible for the care and
custody of the samples collected until they are transferred.   All
samples will be accompanied by a chain-of-custody record and will
be custody-sealed.  This procedure includes use of a custody seal
such that the only access to the package is breaking the seal.
When  samples are  transferred  from one  individual's  custody to
another's, the individuals relinquishing and receiving will sign,
date,  and note the time on the record.  This record documents the
transfer of custody of samples from the sampler to another person
and, ultimately, to a specified analytical laboratory.

Shipping containers will also be custody-sealed for shipment.  The
seal shall be signed before the sample is shipped.  The chain-of-
custody record will be dated and signed to indicate transfer.  The
original record  will accompany the shipment and a copy  will be
retained by the  sample  collector.   Whenever  samples  are split, a
separate chain-of-custody record will be prepared for those samples
and marked to indicate with whom the samples are being split.  If
samples are being  sent  by common  carrier,  copies of  all bills of
lading or  air bills  must be retained as part  of the permanent
documentation.

References

Bell,  T.A.,  and D.V.  Lightner.    1988.    A  Handbook  of normal/
     penaeid  shrimp  histology.    The  World  Aquaculture Society,
     Baton Rouge, LA.

Berthou,  F.,  G.  Balouet, G. Bodennec,  and M.  Marchand.  1987.  The
     occurrence of  hydrocarbons and histophatological abnormalities
     in oysters  for  seven years following the  wreck  of the Amoco
     Cadiz in Brittany  (France).   Mar. Environ. Res.  23:103-133.

CERCLA.    1988.   Natural Resource Damage  Assessments.  53 Federal
     Regulation 5166 and 9769.
                               B-3

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Haensly, W.E., J.M. Neff, J.R.  Sharp,  A.C.  Morris,  M.F. Bedgood,
     and P.D.  Boem.  1982. Histopathology of Pleuronectes platessa
     L. from Aber Wrac'h and Aber Benoit, Brittany,  France: long-
     term effects of the Amoco Cadiz crude oil spill.  J. Fish Dis.
     5:365-391.

Sparks, A.K.  1985.  Synopsis  of invertebrate pathology excluding
     insects.   Elsevier Publ., New York.
                               B-4

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                            APPENDIX C

              GLOSSARY OF ABBREVIATIONS AND ACRONYMS
ADF&G     Alaska Department of Fish and Game
AFK       Armin F. Koernig Fish Hatchery
AHs       Aromatic Hydrocarbons
AHH       Aryl Hydrocarbon Hydroxylase
ANOVA     Analysis of variance
AP        Alaska Peninsula
A/W       Air/Water
AWL       Age, Weight, Length
CERCLA    Comprehensive  Environmental  Response,  Compensation and
          Liability Act
CH        Coastal Habitat
CI        Cook Inlet
CIK       Cook Inlet/Kenai
CTD       Conductivity/temperature/depth
CWA       Clean Water Act
CWT       Coded wire tag
DEC       Alaska Department of Environmental Conservation
DNR       Alaska Department of Natural Resources
DOA       Department of Agriculture
DOC       Department of Commerce
DOI       Department of the Interior
DOJ       Department of Justice
DBMS      Database Management System
EPA       Environmental Protection Agency
ES        Economic Study
EVOS      Exxon Valdez Oil Spill
FRED      Fisheries  Rehabilitation,  Enhancement  and Development
          Division, ADF&G
F/S       Fish/Shellfish
FWS       U.S. Fish and Wildlife Service
GC-MS     Gas chromatography-mass spectrometry
GOA       Gulf of Alaska
KAP       Kodiak Archipelago/Alaska Peninsula
KP        Kenai Peninsula
LCI       Lower Cook Inlet
LKP       Lower Kenai Peninsula
MFO       Mixed function oxidase
MLLW      Mean lower low water
MM        Marine Mammal
NIOSH     National Institute of Occupational Safety and Health
NMFS      National Marine Fisheries Service
NOAA      National Oceanic and Atmospheric Administration
NPH       Naphthalene
NPS       National Park Service

                               C-l

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                           APPENDIX C

              GLOSSARY  OF ABBREVIATIONS AND ACRONYMS
NRDA      Natural Resource Damage Assessment
NSO       Nitrogen-sulphur-oxygen
OSSM      On-Scene Spill Model
PED       Potential egg deposition
PHN       Phenanthrene
PI        Principal Investigator(s)
PWS       Prince William Sound
PWSAC     Prince William Sound Aquaculture
QA/QC     Quality Assurance/Quality Control
RPWG      Restoration Planning Work Group
SCAT      Shoreline Cleanup Advisory Team
SSAT      Spring Shoreline Assessment Team
TM        Terrestrial Mammals
TS        Technical Services
USFS      United States Forest Service
VFDA      Valdez Fisheries Development Association
                               C-2

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