Field Operations Manual

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I.   INTRODUCTION
Background
Bight'98

II.    OVERVIEW OF FIELD SURVEY
Sampling Period
Sampling Design
Indicators of Ecosystem Health

III.   DESCRIPTION OF FIELD TEAMS AND ACTIVITIES
Personnel
Station Assignments
Equipment
Chain-of-Command
Weekly Communications
Daily Communication
Important Telephone Numbers

IV.   SAFETY

V.   QUALITY ASSURANCE/QUALITY CONTROL PROCEDURES
Protocol Calibration/Quality Assurance Procedures

VI.   SAMPLING LOGISTICS
Navigation
Sampling Schedules
Station Types
Cruise Log
Site Acceptability Criteria
Site Rejection Strategy

VII.   BENTHIC SAMPLING
Sampling Effort
Van Veen Grab
Grab Sampling Procedures
Criteria for Acceptable Grab Samples
Benthic Data Sheet
Sediment Description
Sample Processing

VIII.   TRAWL SAMPLING
Purpose
Sampling Effort
Collection Permits
Otter-Trawl Specifications
Pretrawl Survey
Trawl Data Sheet
Net Preparation
Trawling
Criteria for Accepting a Trawl
Sample Processing
Fish samples for Bioaccumulation/Biomarker Studies
Biomarkers
Quality Control Procedures
Voucher Collection

IX.   LABELING AND SHIPPING OF SAMPLES AND FIELD DATA SHEETS
Sample Labels/Tracking
Labels
Field Data Sheets
Shipping of Samples
Chain of Custody Forms

X.   CONTINGENCY PLANS
Purpose
Adverse Weather Conditions
Station Inaccessibility
Lost Gear

XI.   WASTE DISPOSAL
Routine Garbage
Detergent Washes
Chemicals
Fish Waste

XII.   FIELD DATA BASE MANAGEMENT

XIII.   BIGHT''98 PROGRAM ORGANIZATION

XIV.   LITERATURE CITED

XV.   APPENDICES
   Appendix 1:   Bight'98 Station location charts
   Appendix 2:   Bight'98 Field Sampling Sampling Organizations
   Appendix 3:   Bight'98 Sample Processing Analytical Laboratories
   Appendix 4:   Equipment and Supply lists
   Appendix 5:   Sampling Vessel Specifications
   Appendix 6:   Data Sheets
   Appendix 7:   QA/QC Data Sheets
   Appendix 8:   Organization Contacts
   Appendix 9:   SCCWRP Sample Shipping Information

The Southern California Bight Pilot Project (SCBPP) was conducted in 1994 to begin addressing regional monitoring concerns. This project was the largest regional survey of environmental conditions on the mainland shelf in the Southern California Bight (SCB). It capitalized on the interest and cooperation existing in southern California and the resources available in current monitoring programs to develop an integrated and coordinated regional monitoring program that addresses the needs of the participating local, state, and federal agencies, and provided new management information. The SCBPP provided a much needed first "snapshot" of the state of the SCB. The SCBPP resulted in consistent regionwide data sets for describing pollution exposure and biological resources within the SCB. By sampling 261 sites on the mainland shelf of the SCB, an assessment was made of pollutant exposure, the status of biological resources and species diversity, and the presence of marine debris in the SCB.

The Southern California Bight 1998 Regional Marine Monitoring Survey (Bight’98) will continue the development of regional scale management information and will follow the general plan of the 1994 SCBPP. Bight’98 includes a substantial increase in the number of participants, who will provide additional resources thereby increasing the scope of the project and allowing the introduction of new indicators such as a shoreline microbiology component and analysis of biomarkers in fish. Increased participation will also allow for an expansion of the number of sample sites and habitats (e.g. San Diego Bay, Catalina Island, the Channel Islands, and historically sampled reference sites).

The proposed goal of Bight’98 is to make an assessment of the current environmental status of the mainland shelf and shoreline along the southern California. To accomplish this objective, a generalized question has been posed that can be paraphrased as "What is the extent and magnitude of change in an indicator measured in the SCB?". Subpopulations, or strata, of interest have been identified that will allow us to answer other related questions: 1) Is the degree of change similar throughout the SCB, or is it more severe in particular areas?; 2) Can changes be associated with identifiable sources of pollution, such as municipal wastewater outfalls, rivers, or harbors?; and 3) Are the associations identified the same throughout the SCB?

The Bight’98 Survey is organized into three technical components: 1) coastal ecology, 2) microbiology, and 3) water quality. Separate microbiology and water quality surveys will be conducted throughout the following year which will supplement the Bight’98 summer field study. These studies will be conducted according to protocols described in separate documents produced by the microbiology and water quality working groups. The Bight’98 coastal ecology technical component’s field sampling element will be conducted in July and August of 1998. The purpose of this document is to provide detailed instructions on all field sampling methods that will be used to conduct this study.

The index period for the Bight’98 study will extend from July 13, to September 4, 1998.

The Bight’98 study will use a probability-based sampling design developed by EMAP that combines the strengths of systematic and random sampling. This sampling design consists of a grid of tessellated hexagons with a station selected at random within each hexagon. Sampling can be intensified in areas of special interest by decreasing the size of the hexagons, thereby increasing the number of hexagons in an area.

Bight’98 has identified 15 different strata of stations that will be sampled in this survey. These strata are classified as follows: Channel Islands (California Current influenced), Channel Islands (Davidson Current influenced), Catalina Island, shallow offshore (6 to 30 m), mid depth offshore (30 to 120 m), deep offshore (120 to 200 m), river mouths, small POTW outfalls, large POTW outfalls, historical sampling sites (30 m depth), historical sampling sites (60 m depth), marinas, port/industrial, other bays or harbor areas, and San Diego Bay.

The primary goal of the Bight’98 project is to provide an assessment of overall ecosystem condition of the SCB. To accomplish this goal, the following indicators of ecosystem health will be examined:

• Water-column -- microbiological indicators, temperature, salinity, depth, dissolved oxygen (DO), and transmissivity
• Benthic -- sediment characteristics, sediment contamination, infaunal assemblages, and sediment toxicity;
• Demersal fish and invertebrate assemblages, and fish gross pathology, biomarkers, and bioaccumulation
• Marine debris (including plastic, lumber, vegetation, glass, etc.)

All field sampling will be conducted by personnel knowledgeable in field sampling (e.g., trawling, benthic sampling, and hydrocasting). Teams of field personnel will be on each research vessel participating in the sampling effort. These groups will vary in size depending on which organization is doing the field sampling. The main requirements are that the personnel on board the vessel:

• Have a good working experience with the different types of sampling devices
• Are able to troubleshoot problems when they arise
• Have the knowledge and experience necessary for conducting the field collection and analysis of benthic invertebrates and sediments, and trawl-caught demersal fish and megabenthic invertebrates

Each organization's vessel will have its own Boat Captain and crew. A Chief Scientist will be in charge of the scientific crew and will have the final decision on whether to abandon or sample a station. This person is also responsible for assuring the quality of the data and making sure that all data from a sampling cruise is delivered to appropriate information processing personnel in a timely manner.

The mainland shelf of the Southern California Bight will be divided among the participating organizations according to the level of effort contributed by each. The number of stations to be sampled by each organization is summarized in Table 1. Maps and coordinates of the stations to be sampled by each organization are located in Appendices 1 and 2, respectively.

All groups or organizations involved in the sampling program will provide their own research vessel, crew, Van Veen grab, otter trawl, and any other equipment necessary to complete the sampling assignment. A list of equipment used during the survey is provided in Appendix 4. Characteristics of each vessel to be used in this survey are listed in Appendix 5.

Each organization will have a minimum of two modified Van Veen grab samplers. Grab specifications are given in Section 7.

Each organization will have a sufficient number of 7.6 m (headrope) trawl nets and sets of otter boards (doors) available. Net and door specifications are given in Section 8.

Cellular phones are recommended, but not required, to facilitate communication between the Chief Scientist on the sampling vessels with land based Bight’98 project personnel. Vessel cellular telephone numbers are listed in Appendix 5.

The following chain of command is recommended to avoid confusion, identify responsible parties, and to ensure that proper sampling protocols and information flow are followed by each Organization:

• Each organization will establish the chain-of-command for their own vessels. The Chief Scientist is directly responsible for all field activities conducted by the crew.
• Any changes that are made to the established logistical plan that are outside of the jurisdiction of the Chief Scientist will be communicated to the Chief Scientist by either the Field Coordinator (Tim Rothans) or the Project Manager (Steve Weisberg). The teams will accept technical direction from no other authority. All changes to the sampling plan that occur during the field surveys must be documented.
• All technical matters, such as equipment problems, questions regarding station locations, sampling schedules, etc., will be addressed to the Field Coordinator by the Chief Scientist AS SOON AS POSSIBLE.
• For participants that are subject to field audits prior to the sampling period, the Quality Assurance/Quality Control (QA/QC) Specialist will inform the Chief Scientist of any areas where field operations need to be adjusted to meet Bight’98 protocol or where there are problems in taxonomic identification. The Chief Scientist will be expected to take the appropriate action to correct the situation.

Each week a representative from each participating organization will fax to SCCWRP (ATTENTION: Larry Cooper) a schedule of proposed sampling days during the upcoming week, operations to be conducted, and general areas where sampling is likely to occur. This information is needed by QA/QC auditors who will be participating on different cruises on different days. Prior to a QA/QC audit, the auditor will contact each Chief Scientist to verify that the proposed schedule is still in place.

At the completion of every sampling day, or when the vessel returns to port, each participating organization will fax to SCCWRP (ATTENTION: Larry Cooper) a listing of sampling that was conducted that day on the designated sample tracking forms (Appendix 6). A separate form will be submitted for trawls and benthic sampling. This information is needed to provide a summary of progress of the survey as needed to the Bight’98 Steering Committee and to determine the need for follow up sampling by a contractor.

The names and phone numbers of appropriate personnel and emergency services are listed in Section 13 and Appendix 8. If a particular individual cannot be reached at the listed number, the caller should call SCCWRP, where an attempt will be made to provide a phone number where the individual can be reached.

Collection of samples in field surveys is inherently hazardous and this danger is greatly compounded in bad weather. Thus, the safety of the crews and equipment is of paramount importance throughout the project. Each person working on board a vessel during the project should take personal responsibility for their own safety.

Many accidents at sea are preventable. Safety awareness by the Boat Captain and all crew members is the greatest single factor that will reduce accidents at sea. Each survey crew should follow established rules and provisions within their respective Organization's safety program. Sampling should be canceled or postponed during hazardous weather conditions. The final decision is made by the Boat Captain, who is responsible for the safety of everyone on board. As with any field program, the first priority is the safety of the people on board, followed by the safety of the equipment, and the recovery of the data.

The Bight’98 survey will be conducted cooperatively by a number of organizations which routinely monitor the marine environment according to established protocols. It is important to the success of the Bight’98 study that comparable data are collected by each Organization. This Field Operations Manual will provide information on how field operations will be conducted to meet this requirement. Chief Scientists and Boat Captains will be instructed on the field procedures to be followed during the survey and they, in turn, will instruct their field personnel on the proper procedures for the survey.

The Chief Scientist of each organization is responsible for distributing the Bight’98 Field Operations Manual to all field personnel ensuring that their staff understands and uses the protocols detailed in the manual.

Chief Scientists and Boat Captains of all organizations participating in the survey will be required to attend a protocol calibration meeting, conducted prior to the survey on June 24, 1998. The goals and objectives of the Bight’98 were discussed at this meeting as well as the responsibilities of the Chief Scientist and Boat Captains during the Bight’98 survey. Each participating organization has been provided with a workplan and Field Operations Manual for Bight’98 and has been instructed on field procedures to be used during the survey, including proper entry of data on field data forms. The meeting emphasized decision making procedures for determining whether a station should be abandoned and whether a sample is acceptable. Lines of communication within the project and QA/QC activities occurring on the boat during the survey was also discussed.

The Chief Scientist of each organization will train their field personnel, as needed, on the field operations to be conducted during the survey. It will be the responsibility of the Chief Scientist of each organization to review the Workplan and Field Operations Manual with their field crews and to ensure that they understand that these procedures must be followed during the survey. It is also the Chief Scientist’s responsibility to train their field crews, as needed, on operations to be performed. Personnel that cannot perform an operation as required by the project will not participate in that operation.

The participation of several different vessels and field sampling teams in Bight’98 requires that uniform procedures be followed in the field to ensure high quality samples and consistent results. Field personnel will be provided with the Bight’98 Field Operations Manual and instruction on sampling procedures, application of sample acceptance criteria, sample processing, and use of field data forms. All participants are expected to understand and properly carry out all steps in the collection, screening, relaxation, and fixation of infaunal samples; and the subsampling and handling of sediment chemistry and toxicity samples.

An attempt will be made to ascertain field sampling capability by means of field audits conducted by the Field QA Specialist, or designee, prior to sampling for the Bight’98 study. These audits will be conducted by representatives of the organizations who participated in the SCBPP (CLAEMD, LACSD, MEC, CSDMWWD, and SCCWRP) and have adopted the field methods prescribed in that survey as standard operating methods for routine monitoring. The following organization pairs have been established to coordinate and complete the QA audits: SCCWRP and CINMS and WIES; MEC and SeaVentures; CSDMWWD and SPAWAR; LACSD and MBC; and CLAEMD and ABC.

During the field audits, the QA Specialist will provide corrective instruction as necessary. The Field QA Specialist (or designee) may also conduct subsequent audits on benthic sampling procedures during the Bight’98 survey to ensure that sampling is conducted in a uniform manner and all required information is recorded by all field crews.

The goal of the Bight’98 survey is to collect grabs at all sites. However, a Measurement Quality Objective (MQO) of 90% has been established for completeness for the collection of the benthic samples. This completeness goal is established in an attempt to derive the maximum statistical power of the sampling design and was not set at 100% in recognition that some sites will be difficult, if not impossible, to sample with a Van Veen grab. Nevertheless, field crews are expected to strive to collect samples at 100% of the stations.

Demersal fish and invertebrate assemblage data (species identification, enumeration, biomass, and length) are significantly influenced by the collection methods. Therefore, strict adherence to prescribed sampling protocols is critical. Fish catches are influenced by gear type and deployment, tow duration, and towing speed. All organizations collecting samples in the field must use standard nets and follow standard trawling procedures to ensure that comparable samples are collected. Field personnel will be provided with the Bight’98 Field Operations Manual. The Chief Scientist of each organization is responsible for ensuring that their staff understands and uses the protocols as detailed in the manual.

Several QA/QC activities will help to ensure the quality of the trawl survey data. These include intercalibration checks of equipment, sample processing, and taxonomic identification. Trawl equipment, deployment, and sample processing protocol will be checked in presurvey (for new organizations) and in-survey visits to each vessel by the QA/QC auditors. The auditors will assure that methods used are those prescribed in the Field Operations Manual.

The QA/QC auditor will check trawling procedures and equipment to ensure that trawling is conducted in the same way by each organization and that the appropriate data is recorded on a Field QA/QC Checklist (Appendix 7). The auditor will check to make sure that the net is rigged properly, that the appropriate data are recorded, that the trawl is deployed and retrieved properly, and that the catch is properly processed. A check will also be made to see that the scales are calibrated at the start of each day, that other pertinent processing equipment are on board, and that processing is conducted according to methods described in the field manual (Appendix7). The Chief Scientist will be notified of the audit results so that any problems can be corrected prior to sampling.

Presurvey field audits will be conducted by a QA/QC auditor on vessels of new participating organizations prior to the survey to assess equipment, vessels, standard protocols, and to instruct the crew, as needed, on the trawling procedures described in the manual. Precruise QA audit data will be recorded on a Field QA/QC Checklist (Appendix 7). The five primary agencies who participated in the SCBPP (CLAEMD, LACSD, OCSD, CSDMWWD, and SCCWRP) have adopted the field methods prescribed in that survey as standard operating methods for their routine monitoring, and, therefore, will not be audited prior to the survey. These agencies will provide required measurements or descriptions of equipment and vessels to be used in the Bight’98 survey.

Lead Bight’98 fish and invertebrate taxonomists will be designated prior to the sampling period. In addition, each organization will identify lead fish and invertebrate taxonomists that will participate in their part of the survey. These individuals must have the required expertise in field identification of trawl-caught fishes and/or invertebrates of coastal southern California at depths of 5-200 m. They will be responsible for providing accurate identifications of species collected during the survey.

While it is expected that the lead taxonomists of each organization have a wide range of knowledge of the common caught trawl species, it is not expected that all persons making field identifications know all species. It is, therefore, very important that each individual realizes their taxonomic limitations and that any guessing be avoided when it comes to finalizing an identification. An error made in the identification of an organism may result in an irretrievable error in the data base due to the fact that most of the organisms that are identified in the field are returned to the sea. If no one on board knows the identity of a specimen, that specimen will be returned to the laboratory for final identification. Once the final identity of any specimen has been ascertained in the laboratory, that change will be noted on either the trawl fish, or the invertebrate species sheets.

Several QA/QC activities will help to assure accurate taxonomic identification of fishes and invertebrates during the survey. Three presurvey QA activities will help to ensure the accuracy of taxonomic identifications made during the survey by providing training and intercalibration among organizations:

1) Prior to the survey, a list of recommended taxonomic identification aids will be distributed to participating organizations. Lists of trawl-caught fish and invertebrate species for southern California will also be distributed. A reference collection of voucher specimens of species collected in the SCBPP is available at SCCWRP for individuals wishing to see species likely to be encountered in the survey. In addition, it is recommended (but not required) that field taxonomists attend one or more of the presurvey information transfer meetings given at SCCWRP on the identification of expected trawl species.

2) Taxonomists from participating organizations will participate in a field presurvey intercalibration exercise to ensure that each organization makes similar identifications on common species.

3) Taxonomists from each organization will also participate in a presurvey intercalibration exercise to assess the probability of taxonomic error in the survey. In this exercise, each organization will identify specimens of representative trawl-caught species in a bucket of fish and in a bucket invertebrates passed between organizations. A numbered tag will be attached to each organism, so that the identification can be checked against the correct specimen. This exercise will focus on identification errors. Correct identifications or "Return for Further Identification" are acceptable. The latter indicates that the specimen would be returned to the laboratory (where additional information or expertise can be found) for final identification. Organizations with more than 5% misidentifications (fish and invertebrates combined) will redo the exercise with a new bucket of organisms.

Measurement Quality Objectives (MQOs) for the trawl fish and invertebrate sampling effort are defined in terms of accuracy, precision, and completeness. Acceptability criteria have been established for trawl sample collection. The goal of the Bight’98 trawl survey is to collect samples at all designated trawl and tissue stations, to identify all of the organisms correctly, and to obtain accurate counts, measurements, and weights on all species. However, the MQOs will be set at lower values in recognition of the realities of field sampling. Because some stations may occur on rocky bottom, the MQOs for the study completeness objective for trawl sample collection will be 90%. Of the samples collected, 100% will be processed, identified, counted, measured, and weighed. Accuracy expectations for the crew performance is 95% for identification and 90% for counting, lengths, and biomass. The precision objectives are 90% for fish lengths and within 0.2 kg for biomass.

The benthic and trawl surveys may begin July 13, 1998. All field work may be completed in the order that each Organization sees fit, as long as the survey is completed by September 4, 1998.

All samples will be collected between sunrise and sunset. Otter trawl samples must be collected between one hour after sunrise and one hour before sunset. Some types of samples can be processed after dark if necessary but sample collection must be done during the day.

Fifteen different strata of stations will be sampled during the survey. These strata are classified as follows: Channel Islands (California Current influenced), Channel Islands (Davidson Current influenced), Catalina Island, shallow offshore (6 to 30 m), mid depth offshore (30 to 120 m), deep offshore (120 to 200 m), river mouths, small POTW outfalls, large POTW outfalls, historical sampling sites (30 m depth), historical sampling sites (60 m depth), marinas, port/industrial, other bays or harbor areas, and San Diego Bay. The specific strata connected to a sampling station is identified in Appendix 2. In the event that relocating a station moves the station into a different sampling strata, the station will still be sampled and the new strata will be noted in the comments section of the field data sheet.

The Chief Scientist will be responsible for maintaining a Cruise Log (Appendix 6), which will record the basic vessel, crew, and tide information along with all relevant activities conducted throughout the sampling day.

The location of each station will be designated in advance as a set of coordinates (latitude and longitude). Upon arrival at the site, the station depth will be determined by fathometer. This will be regarded as the nominal station depth for all subsequent sampling at the station during the survey and will be used for calculating station acceptability if the station must be moved.

Sampling may not be possible at some stations for a variety of reasons (e.g., kelp beds, rocky bottom, falling outside depth range, etc.). The fathometer reading at a station should be examined to determine whether the bottom is unsuitable for sampling. If the station cannot be sampled, the following rules will be followed:

• The station may be moved no more than 100 m (.054 nautical miles) from any assigned coordinate site and + 10% of the nominal depth. No station will be sampled at depths greater than 220 m, or less than 6 m for the offshore stations, or less than 3 m for stations in bays or harbors.
• If, after three attempts to locate a suitable station, the station still falls in an area unsuitable for sampling, the station may be abandoned and the reasons for station abandonment will be recorded.

A sampling site may be rejected if any of the following occurs:

• If the location places the site on land or in an obviously unsuitable location.
• If the site exceeds the depth boundaries (+/- 10 %) established for the project (e.g. < 3 m in bays and harbors, and offshore depths of < 6 m, > 220 m for trawls, and > 132 m for benthics).
• For benthic sites, if suitable substrate is not found after three attempts at the nominal location, and up to three attempts at two other locations.
• For trawl sites, if the fathometer survey identifies unsuitable substrate at three locations, if any equipment is lost or damaged, or if the site is deemed unsuitable by the Chief Scientist, or their designate, for a valid reason.

A total of 404 benthic stations will be sampled during the survey. The total number of stations sampled by each organization and the parameters to be sampled at each of these sites are listed in Table 1 and Appendices 2 and 3.

A 0.1 m2 modified Van Veen grab will be used to collect sediment samples for physical, chemical, and infaunal analysis (Stubbs et al. 1987). This device is manufactured by Kahl Scientific Instrument Corporation, 737 West Main Street, El Cajon, California 92022 (619/444-2158). The grab may be galvanized, stainless steel, or Teflon-coated. All surfaces of the grab must be clean and free of rust. Either single or tandem Van Veen grabs are acceptable.

Prior to deployment, the grab is cocked with the safety key in place. The grab is then hoisted over the side, the safety key is removed, the grab is lowered at 2 m/sec until it is 5 m above the bottom where it is then lowered at 1 m/sec to minimize the effects of bow wave disturbance of the surface sediment. After bottom contact has been made (indicated by slack in the winch wire), the tension on the wire is slowly increased, causing the lever arms to close the grab. Once the grab is back on board, the top doors are opened for inspection. The first grab collected at a site should be used for the benthic infaunal sample. The chemistry and sediment toxicity samples will be collected from subsequent grabs.

Different sediment types (e.g. cobble, gravel, well sorted sands) and localities (e.g. canyons, slopes, rocky areas) may be difficult to sample. Sediments containing rocks create the most common problem by preventing complete closure of the grab by allowing sediment to wash out during retrieval. Some of the Bight’98 sampling sites may occur on these difficult sediment types or localities because of the randomized sampling design. Therefore, if after three unsuccessful attempts at a site and up to three more unsuccessful attempts at each of two other locations (within 100 m distance from and ±10% of the depth of the nominal site), the station should be abandoned.

Criteria For Acceptable Grab Samples

Upon retrieval of the grab the acceptability of the sample must be determined. Acceptability is based upon two characteristics of the sample: sample condition, and depth of penetration. Sample condition is judged using criteria for surface disturbance, leakage, canting, and washing (Figure 1).

An acceptable sample condition is characterized by an even surface with minimal disturbance and little or no leakage of the overlying water. Heavily canted samples are unacceptable. Samples with a large amount of "humping" along the midline of the grab indicating washing of the sample during retrieval are also unacceptable. While some humping will be evident in samples from firm bottoms where penetration has been poor, this is due to the closing action of the grab and is not evidence of unacceptable washing.

If the sample condition is acceptable, the overlying water is drained off and the depth of penetration determined by insertion of a plastic (rather than metal) ruler vertically along the grab midline and measuring to the nearest 0.5 cm. Sediment penetration depth must be at least 5 cm: however, penetration depths of 7-10 cm should be obtained in silt (fine sand to clay).

Extra caution should be taken to drain the overlying water from the grabs for chemistry and toxicity samples. It is recommended that a siphon be employed for these grabs to avoid disturbance and loss of the surface sediments. The overlying water in grabs intended for infaunal samples may be drained by slightly opening the jaws of the grab and allowing the water to run off, as long as all drained water is captured for screening with the sediments (see Sample Processing below).

If both sample condition and penetration are acceptable, sampling at the station proceeds with the collection of chemistry and sediment toxicity samples from successive grabs. It is required that all of the grabs taken at a station be of similar sediment type and depth penetration. If only one sample of a series of grab attempts is usable, the sediment sample should be kept for infaunal analysis.

The Chief Scientist is responsible for the proper completion of the Benthic Data Sheet (Appendix 6) for each station. The information recorded on this sheet includes date, time of sampling, weather and sea state, navigational position, and other information applicable to the sampling site. For each sample collected (i.e. infaunal, chemistry, toxicity), sampling failures, grab penetration depth, sediment description (type, color, and odor), vessel location, and other sample handling information are to be recorded.

The field description of sediments is required following measurement of penetration depth. The sediment should be characterized as being coarse sand, fine sand, silt or clay, gravel, or of a mixed type. The presence of petroleum tar and shell hash should also be recorded. Obvious odors, such as hydrogen sulfide (the odor of rotten eggs), petroleum, other odors, or a lack of noticeable odors should be recorded. General sediment colors (i.e., black, green, brown, red, olive, or gray) should also be recorded.

After the sample description has been completed, the sediment sample intended for biological analysis is washed from the grab and screened. All wash waters used on the sample are to be filtered in some fashion to prevent the accidental introduction of surface-water organisms. Thoroughly wash the sediment from the grab and transfer it to a sediment washing table for screening.

A means of capturing all water drained from the grab, the grab sample, and the wash water must be used. Typically, a tub (>70 L capacity) is positioned under the grab. The use of a sediment washing table is recommended, but not required. The table is useful in that provides a flat, smooth surface over which to spread and wash the sample, thereby providing a means of gently breaking up the sediment before it runs off the end of the table into the screen box. The screening box must be equipped with a stainless steel mesh with 1.0-mm openings. Wire diameter should be similar to that found in the U.S. Standard 1.00 mm Sieve (0.58 mm). The surface area of screen should be adequate to easily accept the sample without build up. Typical surface areas used in surveys in the Bight are 1500 to 2100 cm2. While washing the sample, control the water pressure to avoid damaging the organisms. Minimize direct application of water from the hose to the material and organisms collecting on the screen.

Once the sample has been washed through the screen, transfer the material (debris, coarse sediment, and organisms) retained on the screen to a sample container. The sample container is to be labeled with an external label containing the station name, sample type, date, and "split number" (i.e. 1 of 1, 2 of 3, etc.) if required. An internal label bearing the same information is placed inside the infaunal samples. This label is written in pencil or indelible ink on 100% rag paper, poly-paper, or other paper of a quality suitable for wet labels. The sample container must have a screw-cap closure and be sufficiently large to accommodate the sample material with a head space of at least 50% of the container volume. A sample may be split between two or more containers. However, each container must have external and internal labels (as described above) with the appropriate "split number" clearly marked. Field crews should have a broad range of sample container sizes available to them with none less than 16 oz (0.47 L) capacity.

Gently remove the material retained on the screen, taking care to avoid damaging the organisms. The sample container should be filled approximately 50 to 70% of capacity with screened material. After the bulk of material has been transferred to the container, closely examine the screen for any organisms caught in the mesh. Remove any organisms with forceps and add them to the sample container. Thoroughly wash the screen box and scrub the mesh before the next sample is screened.

All infaunal samples will be treated with relaxant solution for approximately 30 minutes prior to fixation. Either an epsom salts (MgSO4) solution or a propylene phenoxytol solution (formulations below) may be used for this purpose. Relaxant solutions may be used as the diluent water for the fixative, or may be decanted after exposure and replaced with diluted fixative. If it is used as diluent water, fill the sample container to 85 to 90% of its volume, close the container and invert it several times to distribute the solution. Leave the sample in the relaxant for 30 minutes. After 30 minutes top off the container with enough sodium borate buffered formaldehyde to achieve a 10% formalin solution. Close the container, once again, and invert it several times to assure mixing. Store the sample for return to the laboratory.

If the relaxant solution is not used as the diluent water, the relaxant must be removed from the sample container and replaced with 10% formalin. After the 30 minutes treatment, decant the relaxant from the sample through a screen with a mesh size of 1.0 mm or less. Insure that all animals are removed from the screened and placed in the sample container, then fill the container with sodium borate buffered 10% formalin rather than undiluted formaldehyde. Close the container, invert it several times to assure mixing, and store it for return to the laboratory.Relaxant and fixative stock solution alternatives are as follows:

Epsom salts relaxant solution:	1.5 kg Epsom salts (MgO4@7H2O) per 20 L of fresh water.
Propylene phenoxytol solution:	30 ml propylene phenoxytol to 20 L of seawater.
Buffered formalin solution:	50 g sodium borate (Na2B4O7) per liter of formalin.
Buffered 10% formalin solution:	1 part buffered formalin to 9 parts fresh or salt water.

Following collection of benthic infauna, the next grab(s) will be taken for sediment chemistry samples. Sediment in contact with, or within, 1 cm of the metal sides of the grab will be avoided in all cases, with the exception of grain size, to prevent sample contamination. Sediment samples will be collected from the top 2 cm for grain-size, total organic carbon (TOC), trace metal, and trace organic analyses by randomly subsampling undisturbed surface material with a stainless steel, Teflon-coated stainless steel, or plastic scoop. Samples will also be collected for acid volatile sulfur-simultaneously extracted metals (AVS-SEM) analyses using a plastic or a Teflon-coated metal scoop. At the very minimum, the scoop will be washed with soap and water and rinsed with deionized (DI) water between stations. It may also be rinsed with methanol, followed by acetone, hexane, or methylene chloride between stations but these solutions must be disposed of as hazardous waste. Use of a new scoop with each sample is also acceptable. Samples should be placed in precleaned sample containers provided by the analytical laboratory responsible for the sample analysis. The following container types, samples sizes, and storage requirements should be used with the analytical laboratory supplying all sample containers for all parameters except mineralogy:

• AVS-SEM -- Approximately 200 g of sediment material will be collected at each station with a plastic or a teflon coated metal scoop. Sample containers should be filled as quickly as possible to minimize contact with air. Each sample will be placed into a 250 mL polycarbonate centrifuge bottle without stirring (sample containers to be provided by EPA Newport, Oregon Laboratory). Avoid obvious macroorganisms, sticks, debris, etc. and take samples from at least three locations in the grab, at least 1 cm from any grab wall (if grab is not coated, or if it is teflon coated and in obviously poor condition). Containers are to be filled completely, leaving no head space. Samples should be stored at approximately 40C by placing them on wet ice or in a refrigerator until returned to the laboratory. Do not freeze these samples. Samples may be held for no more than three days before transport to SCCWRP. The inter-laboratory transport time will not exceed 24 hours.
• Sediment Grain Size -- Approximately 100 g of sediment material will be collected at each station by filling one 4-oz (118 mL) whirlpak, plastic, or glass container, taking care to leave an air space at the top. For the samples being analyzed by the City of San Diego, place tape around the snap lock cap to ensure that the sample remains closed during transport. Samples should be stored at approximately 4o C by placing them on wet ice or in a refrigerator until returned to the laboratory. Do not freeze these samples. They should be returned to the analytical laboratory within a week of sampling.
• Total Organic Carbon -- Approximately 200 g of sediment material will be collected at each station by filling one 8-oz (236 mL) plastic or glass container with a Teflon-lined lid, taking care to leave an air space at the top. Samples should be stored at <4o C by placing them on wet ice or in a refrigerator but must be frozen within 24 hours. If frozen, they should be returned to the laboratory within a week. If not frozen, they should be returned to the analytical laboratory within 24 hours. If samples need to be transported, they should be packed in dry ice and shipped via overnight express or a local carrier.
• Trace Metals -- Approximately 200 g of sediment material will be collected at each station by filling one 8-oz (236 mL) plastic or glass container with a Teflon-lined lid, taking care to leave an air space at the top. Samples should be stored at <4o C by placing them on wet ice or in a refrigerator but must be frozen within 24 hours. If frozen, they should be returned to the laboratory within a week. If not frozen, they should be returned to the analytical laboratory within 24 hours. If samples need to be transported, they should be packed in dry ice and shipped via overnight express or a local carrier.
• Trace Organics -- Approximately two 200 g replicates or one 500 g sample of sediment material will be collected at each station by filling two 8-oz (236 mL) or one 16-oz (472 mL) glass containers with Teflon-lined lids, taking care to leave an air space at the top. Samples should be stored at <4o C by placing them on wet ice or in a refrigerator but must be frozen within 24 hours. If frozen, they should be returned to the laboratory within a week. If not frozen, they should be returned to the analytical laboratory within 24 hours. If samples need to be transported, they should be packed in dry ice and shipped via overnight express or a local carrier.
• Mineralogy -- 40 mL will be collected at each station and placed into a whirlpak or other suitable plastic container. Do not freeze these samples. There are no other special temperature holding requirements. These samples will be transported to SCCWRP.

Labeling of sample containers will be the responsibility of the field sampling groups with the following minimum information required on each sample label: station number, sampling date, parameter, split (if required).

Following the collection of sediment chemistry samples, grabs will be taken for sediment toxicity analysis. The sediment collected will be used for three different types of tests: amphipod survival, Qwiklite luminescence, and sediment water interface. Sediment from all toxicity stations will be sampled for the amphipod survival test. Additional sediment from a subset of these stations (203) will be collected for the Qwiklite test. Samples for sediment water interface tests will be collected from 30 stations. Separate sediment samples are required for each of these tests.

As with other parts of the program, sample containers will be provided by the toxicity laboratories performing the required analysis. High-density polyethylene (HDPE) containers will be used for collection of amphipod and Qwiklite sediment samples. Sediment water interface samples will be collected in polycarbonate tubes. All sample containers will be provided by the analytical laboratory. Labeling of sample containers will be the responsibility of the field sampling groups with the following minimum information required on each sample label: station number, sampling date, parameter, split (if required).

Sediment samples for amphipod and Qwiklite tests shall be collected from the top 2 cm by sampling the undisturbed surface material with a scoop. Scoops can be made of plastic (e.g., HDPE, Teflon, polycarbonate) or Teflon-coated material. Scoops should be stored in plastic bags when not in use. Contact with sediment within 1 cm of the metal sides of the grab should be avoided to prevent sample contamination. Sample volume requirements for each test are:

• Amphipod – 2.25 L (minimum of 1.25 L), distributed into two 1L containers and a 250 mL container
• Qwiklite – 125 mL (placed in a 250 mL container)

When taking sediment toxicity samples, the sediment from the grabs should be proportionally distributed into each sample container. A 1.5 L minimum will be required for sediment toxicity analysis, with at least 60 mL in the Qwiklite sample container. The station will be considered completed if 1.5 L of sediment has been collected with 5 or more grabs. If the required minimum 1.5 L has been collected with fewer than 5 grabs, then additional grabs will be taken until either 2.0 L have been collected, or 5 total sediment toxicity grabs have been taken. Sediments will not be homogenized in the field. Each labeled container should then be refrigerated, or placed on wet ice.

Samples to be analyzed by the Organization conducting the collection will be returned to their laboratory by the field crew. Samples to be analyzed by other laboratories will be transported to SCCWRP with a completed chain of custody sheet. The samples will be stored at SCCWRP for later distribution (Appendix 3).

Additional samples will be collected for sediment water interface tests (see Appendix 2) by personnel from the Marine Pollution Studies Laboratory. A grab will be subsampled with a series of small subcores. The subcore will be inserted to the sediment to a depth of 5 cm and a maximum of six subcores can be collected from a grab of sufficient penetration. A minimum grab penetration of approximately 10 cm will be needed to enable collection of a complete set of sample cores. The bottom of the subcore is then closed with a gloved hand for removal, the ends are fitted with caps and then wrapped with parafilm for a final seal. The subcores will be stored upright in an ice chest and to be later hand delivered by MPSL staff to the laboratory. Six

subcores will be collected from 15 stations and 11 subcores collected from an additional 15 stations.

Any sample collection equipment which comes in contact with the sediment should be cleaned between stations with the procedures described above. Collection equipment should be stored in plastic bags when not in use. Alternatively, cleaned "sets" of sample collection equipment may be individually prepared and wrapped in plastic per station prior to sampling.

Samples may be held in the field, or laboratory, on wet ice, or in a refrigerator at 4o C, for no more than three days before transport to the designated toxicity laboratories. The inter-laboratory transport time will not exceed 24 hours. Upon arrival to the analytical laboratory, the samples will continue to be stored at 4oC. Testing should begin within two weeks of sample collection. Chain of custody procedures should be followed throughout the sampling and analysis procedures.

The purpose of trawl sampling is to obtain data on the abundance, biomass, diversity, and disease prevalence of demersal fish and invertebrate assemblages. It is also used to collect fish and invertebrates for tissue contaminant analysis. This information is useful in characterizing possible anthropogenic effects on demersal fish and invertebrate populations. Mearns and Allen (1978) provides a comprehensive description of how small otter trawls should be designed and used for conducting biological surveys in coastal waters.

A total of 378 trawl stations will be sampled during the survey. The total number of stations sampled by each organization and the parameters to be sampled at each of these sites are listed in Table 1 and Appendix 1, respectively

The local office of the California Department of Fish and Game (CDFG) (San Diego, 619/237-7311; northern area, 562/590-5132) must be contacted prior to collecting fish and invertebrates. The caller will be asked for his or her name; scientific collector's permit number; date, time, and area of sampling; type of gear to be used; vessel size, color, and CF number or documentation; number of persons in party; and what organisms will be collected.

The permit must be on-board during sampling and must be presented to any CDFG warden or personnel who request to see it.

A semiballoon otter trawl will be used to collect epibenthic invertebrates and demersal fish. Net dimensions are the following: 7.6-m headrope (25 ft); 8.8-m footrope (29 ft); 3.8-cm (1.5 in) body mesh; and a 1.3-cm cod-end mesh (0.5 in). This net will have 22.9-m (75 ft) long bridles made of 1.0-1.6 cm (3/8 to 5/8 in) diameter rope (e.g., Samson braid). The otter boards (doors) will have a width of 76.2 cm (30 in), height of 50.8 cm (20 in), and a suggested weight of 15.9 kg (35 lb). Slight deviations (< 10%) from these dimensions are acceptable. The door chains should be 5-mm (3/16 in) in diameter and should have the following numbers of links: front top -- 12; front bottom -- 11; back top -- 17; back bottom -- 16. At least 700 m (2296 ft) of wire is necessary to obtain a trawl at a depth of 200 m.

Trawl gear is likely to be lost if the trawl is hung up on bottom obstructions and replacement of nets can be costly. The bottom along a trawl course at a previously unsampled station can be examined by a fathometer. A pretrawl survey can enable the navigator to avoid uncharted reefs and other obstacles that may cause damage to trawl gear. This survey should always be conducted at a new sampling site to determine whether the station is acceptable, or it should be abandoned.

The pretrawl survey should follow the expected trawl course along the isobath and the fathometer reading will be examined for rocks and other obstacles. If the first run indicates that the site is unacceptable, another survey will be conducted within 100 m and 10% of the depth of the original site. If this attempt is unsuccessful, a third attempt will be conducted at a different location using the same protocols. If after three unsuccessful surveys, the site will be abandoned.

The Chief Scientist is responsible for ensuring that the proper data is logged on the Trawl Data Sheet (Appendix 6). The information recorded in the log includes water depth, length of tow wire used, times and coordinates (latitude and longitude) for net on the bottom and end of trawl (beginning of trawl retrieval); coordinates for net over and net on deck may also be recorded. Any anomalous conditions such as rocky bottom, rocks in the catch, and torn net will also be recorded in the log.

The trawl should be properly prepared prior to trawling so that the net can be deployed in an orderly and safe manner. The net is laid out and stacked on the stern of the vessel in the same configuration that it will fish, with the cod-end to the stern, the floats up, and the footrope down. The trawl should be checked to make sure that the cod-end is tied, that the doors are connected properly to the leg lines, and that the bridles are connected to the doors and tow wire.

The station coordinates must lie within the course of the trawl. Trawls will be towed along, rather than across, isobaths. While the vessel is underway the Boat Captain will order that the net be placed in the water. It is important that the floats skim the surface and that the net is not entangled while deploying the bridles. This small step could mean the difference between a successful or unsuccessful trawl. The bridles should be paid out by a person(s) on each side of the net, being careful to stand outboard of the bridle lines.

Use of the proper scope (i.e., length of wire paid out versus the water depth) is important to conducting successful trawls. After the net touches the bottom, a sufficient length of hydrowire (towing wire) should be deployed to ensure that the net is pulled from a horizontal rather than a vertical position. Insufficient scope will prevent the net from consistently fishing the bottom and will result in a no-catch, or a short-catch situation. In general, the required scope declines with increasing depth because the additional weight of the hydrowire enhances the horizontal component of the towing forces (Table 2).

These scopes are for 0.6 cm (0.25 in) to 1.0 cm (0.38 in) tow wire.

Once on the bottom, the net is towed for 10 min (5 to 10 min in bays and harbors) at a speed over ground of 1.0 m/sec or 1.5 to 2.0 kn as determined by DGPS, a distance equal to about 600 m (300 to 600 m in bays and harbors).The distance covered during a trawl along with the known width of the net can be used to determine the area of bottom sampled. Trawl speed and duration will be recorded on the Trawl Data sheet.

At the end of 10 min, the net is retrieved and brought on-board the vessel. The cod-end is opened and the catch is dumped into a tub or holding tank where it can then be processed by the scientific crew.

If the trawl is retrieved with little or no catch, its acceptability will be evaluated according to whether the trawl was conducted properly. A trawl is conducted properly if the proper depth, scope, speed, and distance or duration are maintained, if it is not fouled (net tangled), and if there is some evidence (e.g., rocks, benthic invertebrates, benthic fish) that the net was on the bottom. If any of the trawl procedures were not followed, if the net was fouled, or if there was no evidence of contact with the bottom, the trawl will be considered unacceptable and another trawl will be conducted at that site.

If the net is torn sufficiently to allow escapement during the course of a trawl, the station will be abandoned. If the trawl hangs up on the bottom, the site can be resampled or abandoned at the discretion of the Chief Scientist. If retrawling that station proves unsuccessful after another two attempts, the site will be abandoned.

The trawl catch will be sorted on deck into containers. Initially the catch will be rough sorted into major categories (e.g., urchins, shrimp, other invertebrates, flatfish, rockfish, other fishes). The categories used are not important but it is more efficient to sort into rough categories before identifying to species. Debris should also be sorted into containers for processing.

An attempt will be made to identify all trawl-caught organisms in the field using the recommended taxonomic keys and field guides. If unresolvable conflicts arise in the field over the identification of an organism, a note will be made on the appropriate data sheet and the organism will be returned to the laboratory for identification. When the identification has been resolved, the correct identity of the species will be recorded on the original data sheet. If the laboratory identity differs from that recorded in the field, the original name should be crossed out with a line. Do not erase the original name.

Under no circumstances should an organism be discarded if the identity is questioned. Each organization must know its own limitations in identifying organisms.

All fish will be identified, however, only invertebrates meeting specific criteria will be identified. There are likely to be many small infaunal and pelagic species that are incidental to the trawl catch, but only organisms that are greater than 1 cm in any dimension will be identified. Colonial and pelagic organisms will be noted, but not enumerated and infaunal organisms will not be documented. The presence of obvious free-occurring fish parasites, such as leeches or cymothoid isopods, will also be noted.

A recommended list of field guides and taxonomic aids for identifying fish and invertebrates will be distributed to all of the participating organizations prior to the survey. The most basic and comprehensive guides for fish are Miller and Lea (1972) and Eschmeyer et al. (1983). Allen (1977) provides information for identifying juvenile rockfishes (Sebastes spp.) and Kramer et al. (1995) provides information for identifying flatfishes. Generally, there are no widely comprehensive guides to the epibenthic invertebrates.

Either common or scientific names of fish may be used in the field. Only standard common and scientific names of fishes given in Robins et al. (1991), or a list of California fishes (Allen in prep.) will be acceptable. Scientific names of invertebrates will be used in the field (SCAMIT 1998).

Each organization should have a kit containing a variety of tools which will aid in field identification. This kit should include forceps (small with sharp points and large with blunt

points); a hand lens; dividers or calipers; dissecting needles; scalpel with scalpel blades; probe; and plastic ruler in millimeters.

All fish will either be measured using a measuring board or, for very large specimens, using a meter stick or tape measure. A measuring board typically consists of either a flat or trough shaped board with a part of a meter rule embedded in the base. A smaller board is attached perpendicular to the zero-end of the meter stick. Trough-shaped boards are useful in keeping groups of fish on the board during measurement. Centimeter size-classes are marked along the side of the measuring board with the number of the size class marked next to the appropriate centimeter.

During measurement, the anterior portion of the fish is pushed up against the board at the zero-end of the measuring rule. Maximum (board) standard length (i.e., anterior tip of head to base of caudal fin) will be measured on bony fishes (Figure 2) and total length will be measured on cartilaginous fishes; wingspan will also be measured for stingrays because the tips of their tails are frequently broken off. The length of all fish specimens will be reported in size classes of 1 cm intervals (Mearns and Allen 1978). The first centimeter size class (size class number 1) extends from 0.1 to 1.0 cm; size class 2 extends from 1.1 to 2.0 cm, and so forth (Figure 3).

All species will be recorded on the Trawl Fish Species Data Sheet, or Trawl Invertebrate Species Data Sheet (Appendix 6). For species with 10 or fewer individuals, measurements (e.g., size-class 8) will be written on the Trawl Fish Species Data Sheet, separated by commas (Appendix 6). For species with greater than 10 individuals, the size classes will be recorded on the Trawl Fish Size-Class Sheet (Appendix 6) while only the names, the weights and the total number of individuals will be recorded on the Trawl Fish Species Data Sheet.

An attempt should be made to size-class all fish. For the rare occasions when size classing is not possible (e.g., a huge catch of a single species), a subsample of several hundred fish should be measured and the reason for deviating from prescribed procedures should be documented. (Note: Catches of greater than 2,300 individuals of a single species have been measured in past surveys). Lengths of invertebrate species will not be measured.

At a minimum, each Organization should have a range of spring scales that are capable of weighing to the nearest 0.1 kg, and a tare bucket with holes through the bottom or another suitable container (e.g., net bag) on board. Spring scales should be calibrated on a daily basis using a standard set of at least three weights. Tare buckets should be washed periodically to remove slime.

The biomass of each species will be measured with a spring scale. Species with a biomass greater than 0.1 kg will be recorded to the nearest 0.1 kg. The tare container will be weighed while empty and this weight will be subtracted from the weight of the gross weight (species plus tare container) to give the weight of the species (net weight). Tare and gross weight can be recorded on the data sheet but are not required. Small species weighing less than 0.1 kg will be set aside and weighed together to provide a composite weight. Composite weights greater than 0.1 kg will be recorded to the nearest 0.1 kg. Composite weights of less than 0.1 kg will not be rounded; they are to be recorded as "<0.1 kg". There will be one composite weight for fish and one composite weight for invertebrates. These weights will be used in calculating the total biomass of the catch.

Large organisms may be weighed individually. Individual weights of smaller specimens may also be collected using a range of scales capable of weighing to the nearest 0.1 g.

Debris weights should be quantified on the Trawl Debris Data Sheet (Appendix 8?) by classifying specific types identified on the data sheet by quantity categories: trace (<0.1 kg); low (approx. 0.1-1.0 kg); moderate (approx. 1.1-10.0 kg); and high (>10.0 kg).

Total numbers of fish will be tallied following the measurement of lengths. The total number of each species (including those size-classed) should be recorded on the Fish Species List and Fish Size-class Data Sheets.

Most invertebrates will be enumerated following identification. If the number of individuals in particularly abundant species make individual counts impractical, the total number will be estimated from the total biomass of the species. A known number of representative individuals will be subsampled from the catch and weighed. The remainder will then be weighed and the total number of individuals will be calculated (i.e. divide the total biomass by the subsample weight).

Presence of trawl debris should be noted on Trawl Debris Data Sheets (Appendix X) by identifying specific debris types and estimating quantities.

During the identification and measurement procedures, fish and invertebrates will be examined for gross pathology. This involves scanning an individual organism for anomalies and noting any observed pathology for fish (by abbreviation) next to the length of organisms during measurement on the appropriate data sheet. Cooper (in prep.) provides information for identifying anomalies in local fishes and invertebrates. The following anomalies will be noted for fish:

• Fin (and tail) erosion
• Tumors
• External parasites (e.g., copepods, isopods, leeches)
• Color anomalies (ambicoloration, albinism) (Mearns and Haaker 1973)
• Skeletal deformities
• Lesions
• Other anomalies

An observation should be noted next to the individual length on the Fish Species Data Sheet.

For invertebrates, burnspots and other anomalies will be noted in the comment section of the Trawl Invertebrate Species Sheet (Appendix 6).

Fin erosion can be found on the dorsal, anal, and caudal fins of flatfishes, and on the lower caudal fin and pelvic fins of bilaterally symmetrical fishes. Tail erosion occurs on the top and bottom of the caudal fins of bilaterally symmetrical fishes. Tumors can be smooth and rounded (angioepithelial nodules) or furrowed (epidermal papillomas). Externally obvious copepod parasites occur on the eye, fins, or body of fish. Cymothoid isopods occur in the gill cavities of fish or on the body; they often fall off. Leeches occur on the body of some flatfishes. Skeletal deformities include crooked backs, snub noses, or bent fin rays. Lesions include sores that do not appear to be due to net damage. Burnspot disease is found on crabs and some shrimps; its lesions resemble cigarette burns.

Representatives of fish and invertebrates with each disease or parasite should be vouchered. All fish with tumors, fin erosion, and lesions, as well as all invertebrates with burnspot diseases will be returned to the laboratory and vouchered.

Accidental omissions can be occasionally made if a bucket of organisms is not processed. This can be avoided by attaching a colored rubber tag (made of a square with a slit in one side) to the handle of each bucket to indicate a particular stage of processing. For instance, different tags can represent that the bucket is ready for identification, measurement, weighing, preservation, or discarding. As the bucket progresses to the next stage, the current tag can be pulled off and a new tag can be added. This procedure is not necessary for small catches but may be helpful when catches are large. Tags with commonly caught species names can also be temporarily attached to buckets to facilitate sorting and processing.

Field personnel are likely to encounter a variety of organisms that are potentially harmful. The California scorpionfish (Scorpaena guttata) has venomous fin spines with which even a small wound can cause severe pain throughout much of the body. Handle this species with leather gloves or pliers. Hot water, meat tenderizer, or ammonia should be applied to any wound resulting from being stuck by a spine. Heat is effective in breaking down the protein in the venom. Several other species of rockfishes and the spotted ratfish (Hydrolagus colliei) also have mildly venomous spines which can cause a burning sensation. The round sting ray (Urolophus halleri), the California butterfly ray (Gymnura marmorata), and the bat ray (Myliobatis californica) all have venomous spines on their tails.

Pacific electric ray (Torpedo californica) can emit a strong electric shock. When handling this species, hold it by the tail. Do not grasp the disk with both hands!

The Pacific angel shark (Squatina californica), the spiny dogfish (Squalus acanthias), the spotted ratfish, the Pacific electric ray, and the California halibut (Paralichthys californicus) can give painful bites.

Care must also be taken in handling the blueleg mantis shrimp (Hemisquilla ensigera). This species can inflict severe cuts with its raptorial appendages. Care should also be taken in handling any of the large crab specimens.

Fish and invertebrate specimens may be preserved or documented for QC or identification purposes in one of three ways: 1) fixing in buffered formalin-seawater; 2) freezing; or 3) photographing.

Most smaller specimens and fish with fin erosion, tumors, or lesions will be fixed in 10% buffered formalin-seawater. Buffered formalin is made by mixing 50 g Na2B4O7 (sodium borate) per liter of formaldehyde or 5 g per liter of 10% formalin. All specimens with tumors, fin erosions, or lesions should be fixed in this manner. If the specimen is larger than 60 mm, the body cavity should be slit with a scalpel on the right (for most bilaterally symmetrical fish), blind (for flatfish), or ventral side (for dorsoventrally flattened fish, such as rays) thereby allowing the formalin to bathe the internal organs. Note that by convention, bilaterally symmetrical fish are photographed or drawn with their heads facing left and, hence, dissections are conducted only on the right side of the fish. Fish and invertebrates will be placed in plastic bags or plastic jars and fixed in 10% buffered formalin-seawater. Fish should be inserted tail-first into jars so that they can be removed easily without destroying the fin rays or spines.

Fish should remain in formalin for up to a week before being transferred to freshwater. The fish should be left in the freshwater for a minimum of two days, making sure to change the water at least once during that period. The fish should then be transferred to 50% isopropanol (isopropyl alcohol) or 70% ethanol for preservation.

Trawl-caught invertebrates will also be fixed in 10% buffered formalin-seawater and preserved in 70% ethanol. If specimens in formalin are retained at the collecting Organization’s laboratory for more than one week, it is the responsibility of that laboratory to complete the preservation of the specimens (i.e., rinse in water and transfer to isopropanol or ethanol). Specimens that are returned to SCCWRP within one week will be processed for final preservation, then vouchered by SCCWRP.

Larger specimens can be placed in plastic bags and frozen on dry ice or placed in a freezer if sufficiently large containers are not available or if large quantities of fixative are required. These can then be thawed and fixed in the laboratory with 10% buffered formalin solution. However, large specimens with tumors, fin erosion, or lesions should be fixed in the field with formalin rather than frozen.

Small invertebrates (e.g., nudibranchs) may be kept cold in seawater and returned alive to the lab for identification.

Large specimens (and colorful species) of fish and invertebrates can be vouchered in the field by photographing them with color slide film. For larger specimens only, the photographs will document the species of organism; for smaller species, a voucher specimen must be preserved. Photographs should attempt to show the overall appearance of the specimen, as well as any important identifying features. Photographs of unidentified rockfishes should be taken as soon as possible after capture because their color, which is an important taxonomic character, fades during preservation. Bilaterally symmetrical fish and dorsoventrally flattened fish (skates, rays) should be photographed facing left. Flatfishes should be photographed with the eyed side up. The left-eyed species should be photographed facing to the left and the right-eyed species should face to the right. The gill cover should cut the lower profile of the body. If anomalies occur on any specimens being photographed, those features will be also be photo-documented. All specimens should be photographed on a light background with a meter stick along side and a label giving date, station number, and species in large bold letters. Note should be made of character states that can aid in identification (e.g., counts of fin rays, gill rakers, and scales).

Specimens preserved for further identification must be noted on the field data sheet. Note whether the organism is fixed, frozen, or photographed. A photograph log should be kept during the survey, documenting species name, the frame number, the date and the station location of each photograph.

The geographic and depth range of the study precludes selecting a single species. The fish bioaccumulation study will target two categories of fish:

For Category I (sanddab guild) species, a sample composite will consist of six fish of an age class, whereas, for Category II (turbot guild) species, a sample composite will consist of 3-6 fish less than 20 cm (size classes 5 to 20) (Table 3). At each station, composites should be made for each age class of all Category I species and similarly, composites should be made for all Category II species.

Bioaccumulation fish will be collected from the first trawl at a station after it has been processed for assemblage data. If no individuals of Category I species are encountered during the first trawl, no additional trawls will be required and the station will be abandoned (keep any complete composites of Category II species). If six individuals of an age class of a Category I species are collected during the first trawl, tissue sampling at that station is complete and no additional trawling is necessary.

If category I species are collected in the first trawl, but with fewer than six individuals in an age class, additional trawls will be required to complete an entire composite. If six individuals of an age class of a Category I species are collected in the second trawl, tissue sampling at that station will be considered complete and no additional trawls will be required for bioaccumulation. By the end of the third trawl (30 min total trawl time), if an insufficient number of fish are collected for bioaccumulation or biomarkers, no further trawl samples will be collected. No more than 30 minutes of trawling will be required at any of the trawling sites in Bight’98 with the first trawl only at each station processed for assemblage data.

At the end of trawling at a station, all complete composites should be saved for all Category I and II species (i.e. any age of any Category I species with six fish, as well as any Category II species with 3-6 individuals). This will be done because the best combination of species and age classes for chemical analysis will not be known until the end of the survey.

Whole fish age-class composite samples for each species will be packaged in Ziploc plastic bags and placed on dry ice, or in a freezer. Fish samples (composites) will be recorded on the Trawl Cover Sheet (Appendix 6). A label including the date, station, species, and number collected should be placed in each bag and an external label must be stuck on the outside of the bag. In addition, the bag should be labeled on the outside with a felt-tip marker. Any samples to be processed in-house will be transported to the analytical laboratory of that organization. If the analysis is to be run elsewhere, the samples will be shipped to SCCWRP within a week of collection.

Blood and liver/gall bladder samples for biomarker measurement will be obtained from fish collected from two strata: bays and harbors; and Channel Island sites. Fish collection methods will be the same as those used for collection of fish bioaccumulation studies. Because biomarkers degrade rapidly after a fish has died, the fish will be dissected on board by trained personnel soon after collection, and tissues immediately frozen using liquid nitrogen or dry ice.

Fish from San Diego Bay will be dissected on board by SPAWAR Systems Center San Diego (SPAWAR) personnel. The sampling schedule for San Diego bay should be coordinated with Scott Steinert (619-553-5615), who will provide dissection personnel for these samples. Fish from other locations will be dissected by SCCWRP staff or the field crew (after SCCWRP training). Additional samples from selected nearshore reference areas may be collected if samples can be obtained without additional sampling effort (i.e., extra fish are available during a cruise when dissection personnel are present). In order to make the necessary arrangements for biomarker trawls that are to be staffed with SCCWRP personnel, Jeff Brown at SCCWRP (714-894-2222) should be contacted with at least 24 h advance notice.

A minimum of five fish from a single fish species of the sanddab guild will be processed at each site for the biomarkers study. Additional individuals (maximum of 10) will be processed if they are available and time permits. The most abundant target species (at minimum) at that station will be dissected. Fish for the biomarkers study may come from the same trawl as fish collected for the bioaccumulation study, or from additional trawls if insufficient numbers of fish were collected. The same specimens will not, however, be shared between the two studies.

Dissections should be done in an area of the boat that is as free as possible from sources of PAH’s (e.g., diesel fumes). Standard length, species, sex, maturity, time of dissection, and condition of fish will be recorded for each specimen. Blood will be collected by cutting a gill arch, and collecting the resulting blood separately for each specimen in prelabeled plastic microfuge tubes containing PBS/Pronase K/10% DMSO. Dissections will be conducted on boards covered with clean polyethylene sheeting using scissors and forceps. Livers and gall bladders will be removed from each fish as a single unit and put into 19 x 51 mm glass shell vials. All tissues will be frozen on dry ice immediately after dissection. Dissection tools will be cleaned by rinsing with deionized water between samples. Polyethylene sheeting will be replaced as needed. Prior to each day of sampling, instruments will be cleaned by a detergent scrub, followed by acid, methanol, and deionized water rinses.

SPAWAR will keep blood samples from fish collected in San Diego Bay and those collected on SPAWAR vessels in LA/Long Beach Harbor. Blood samples from other locations, and all liver/gall bladder samples will be delivered to SCCWRP within 24 h by field personnel, or shipped by overnight courier. The blood samples will then be sent to SPAWAR.

Shipping information for liver/gall bladder samples:

In addition to the presurvey QA/QC protocols, the following QC activities will check on the accuracy of taxonomic identifications and counts made during the survey:

• During the survey, taxonomic identifications will be checked during at least one visit to each vessel by SCCWRP designated taxonomists. They will observe species identification by each organization in the field and record the data on a Taxonomy QA/QC Data Sheet (Appendix 7). Their duties include rechecking the identifications of at least 25% of the species collected during the day and noting any problems with the identification of pathologies. The Chief Scientist will be informed of any problems and the field personnel will be instructed regarding the appropriate identifications as needed. Each vessel will be expected to have appropriate taxonomic identification aids during the survey.
• During the survey, QA/QC data will also be collected on variability in trawl data collection. On each survey day, the Chief Scientist, or a delegate, will reprocess two randomly selected fish species (of at least 10 fish) that have already been counted, measured, and weighed. These species will be recounted, reweighed, and remeasured. A record of the counts, weights, and lengths of these quality control checks along with the original values will be maintained on a separate size class data sheet and a note will be made in the trawl comments on the original trawl data sheet that the recount was taken.
• Voucher specimens of each species collected by each organization will be preserved and returned to SCCWRP during the survey (see Voucher Collection below). The identification of these specimens will be checked by a qualified taxonomist following the survey to further ensure that identifications were made correctly. Errors in species identifications must be corrected in the data. Anomalies will be verified by a qualified pathologist.

The Bight’98 voucher collection of trawl organisms will be developed during the course of the survey and will be housed at SCCWRP. This collection will document the species taken during the survey and what names were applied to each. It will also document the types of diseases or anomalies found in the gross pathology examinations. It is also recommended that

each organization develop a voucher collection at their organization. The voucher collection will consist of preserved organisms and photographs of organisms.

Each organization will be provided with a list of trawl fish and invertebrate species. For every species caught, each organization will provide at least one representative of that species to the Bight’98 voucher collection. Thus for many species, the Bight’98 voucher collection will contain representatives from all organizations involved in data collection. As species are collected, they will be checked off the list. Each Organization will give specimens to the Bight’98 voucher collection, before including them in their own collection.

Each sample will be identified and tracked by the station, parameter, date sampled, and split number if required. Individual log numbers may be used at the discretion of the sampling organization. Sample log numbers will be handled by SCCWRP for the samples shipped to SCCWRP that are not run by the organization that collected them in the field.

Labels will be printed by the Organization responsible for field sampling prior to the survey and will include, at a minimum, the station number, parameter, date, and split (i.e., 1 of 1, 2 of 3, etc.). Dates will be reported as day/month/year. External labels should be covered with clear postal tape to prevent them from falling off the container if they will not stick on some surfaces.

Benthic data sheets and cruise logs are to be retained by the sampling organization until sampling is completed. Trawl data sheets will be returned to the Organization’s laboratory and held there until all species identifications are complete. Data on species identified in the laboratory must be added to the data sheets and verified within the laboratory. Upon completion of laboratory identifications, original field data sheets are to be sent to SCCWRP with copies retained by the sampling organization. Trawl fish and invertebrate data will be submitted as hard copies and either electronically or on diskettes to SCCWRP as soon as the data sheets are complete.

All sediment, tissue chemistry, and toxicity samples not analyzed by the field sampling organization’s laboratory will be shipped to SCCWRP along with the trawl voucher collection specimens. Most samples do not have to be delivered on the day of collection but can be held for a few days and shipped once a week. All shipping of samples will be the responsibility of the field sampling organizations. See Appendix 9 for detailed SCCWRP shipping information.

Chain of custody forms (Appendix 6) are to be filled out at the end of each sampling day detailing the transfer of samples from the vessel crew to the laboratory, or to delivery personnel. A form is to be filled out for each set of samples that will be transferred to a specific location. The sample and container type is to be included on the form to identify the samples being transferred. This form is to be signed by the crew member transferring the samples and the laboratory staff member receiving them. A copy of the form is to be kept and the original form with signatures will accompany the samples. If samples are shipped by carrier, a copy of the chain of custody form is to be faxed to SCCWRP for tracking purposes.

Any field program can be affected by factors outside the control of the sampling crews. Weather, equipment failure, errors in designating station locations, and accidents can all prevent the field crews from obtaining samples at one or more stations. Contingency plans made in advance of the survey can greatly facilitate decision-making in the field. It is the responsibility of the Chief Scientist to make most of these decisions in the field, based on the protocol described below. If there is any question regarding which protocol to follow, the Field Coordinator should be notified immediately.

It is the responsibility of both the Boat Captain and the Chief Scientist to determine if weather conditions are sufficiently bad to prevent sampling. The Chief Scientist should evaluate all alternatives, such as changing the sampling plan to more protected areas or returning to the prescribed schedule when the weather improves. Every attempt should be made to avoid wasting the entire day. However, the safety of the crew is the number one priority.

Stations can be inaccessible because 1) they were incorrectly positioned on land 2) in water too shallow for the boat, or 3) they cannot be sampled for unforeseen circumstances. If it cannot be sampled, the sampling site will be moved to a location within 100 m horizontal distance from the original site, staying within 10% of the depth of the original site. If it still cannot be sampled, the station will be abandoned. No station should be sampled in less than 3 m or more than 220 m.

Lost gear can potentially have a significant effect on the sampling program. Equipment can be expensive and spares may not be available in a timely manner. Crews should take every precaution against the loss of gear by properly tightening shackles and other connectors.

If important gear is lost, notify the Boat Captain immediately, so he can record the position using the vessel’s navigation system. If possible, deploy a buoy at that exact location so relocation is made easier. Attempt to recover the equipment for a reasonable amount of time. If unsuccessful, use spare equipment (when available) or continue sampling without that particular equipment. Notify the Field Coordinator as soon as possible when equipment is lost.

Proper disposal of all waste is an important component of field activities. At no time will any waste be disposed of improperly. The proper methods of waste disposal are outlined below:

Regular garbage (paper towels, paper cups, etc.) is placed in trash containers on board the boats. It can then be disposed on land in public receptacles or recycled.

Biodegradable detergents are recommended for cleaning the Van Veen Grab, or deck.

Acetone, formalin, hexane and methylene chloride are hazardous materials and should be disposed of by following all appropriate regulations. They should never be disposed at sea.

After each trawl catch has been processed completely, the remaining catch should be disposed of at sea. Discretion should be used. For sampling conducted nearshore or in bays and harbors, return only live fish and invertebrates to the area that trawling occurred. All of the remaining fish should be disposed of offshore. Under no circumstances should fish be given to the public.

A field computer system has been developed for the Bight’98 that includes the forms with all of the fields for all of the field data sheets. This system employs lap-top computers and has an instruction manual for training and reference. Use of the field computer system is optional during the Bight’98 survey.

The data entry screens are identical to the field data sheets. Data can either be entered into the computer while at sea, or it can be taken from the data forms at a later time. Although hard copies of all field data sheets are mandatory, these can either be hand-written, or hard copy print-outs from the computer.

The data entered into each field of the electronic forms is checked automatically by the software and it provides a warning when the data do not fall within an expected range. After entering the data into the field computer system, it will be printed out to hard copy and checked by the Chief Scientist against the original handwritten data sheets. Once the data have been checked, corrected (if necessary) and accepted by the Chief Scientist, the crew will not be granted access to the data any further.

STEERING COMMITTEE Steve Weisberg (Chair) SCCWRP 714/894-2222 FAX 714/894-9699	George Robertson (Co-Chair) Orange County Sanitation District 714/593-7468 FAX 714/962-2591
FIELD SAMPLING/LOGISTICS Tim Rothans (Chair) CSDMWWD 619/692-4914 FAX 619/692-4902	Jim Allen (Co-Chair) SCCWRP 714/894-2222 FAX 714/894-9699
QA OFFICER Terry Fleming U. S. EPA, Reg. IX 415/744-1939 FAX 415/744-1078
CHEMISTRY COMMITTEE Eddy Zeng (Chair) SCCWRP 714/894-2222 FAX 714/894-9699
TRAWL COMMITTEE Jim Allen (Chair) SCCWRP 714/894-2222 FAX 714/894-9699
INFORMATION MANAGEMENT Systems Committee Larry Cooper (Chair) SCCWRP 714/894-2222 FAX 714/894-9699	Jon Bishop (Co-Chair) LARWQCB 213/266-7538
TOXICITY COMMITTEE Steve Bay (Chair) SCCWRP 714/894-2222 FAX 714/894-9699	Tim Mikel (Co-Chair) Aquatic Bioassay & Consulting Laboratories 805/643-5621
BENTHIC COMMITTEE Mary Bergen (Chair) SCCWRP 714/894-2222 FAX 714/894-9699	Dave Montagne (Int. Chair) Los Angeles County Sanitation District 310/830-2400 x402
WATER QUALITY COMMITTEE Burt Jones (Chair) USC 213/740-5765 FAX 213/740-8123	Alex Steele (Co-Chair) Los Angeles County Sanitation District 562/699-7411 FAX 562/695-6139
MICROBIOLOGY COMMITTEE John Dorsey (Chair) City of Los Angeles/Stormwater 213/847-6347 FAX 213/847-5443	Charles McGee (CO-Chair) Orange County Sanitation District 714/593-7504 FAX 714/962-2591

Allen, M. J. 1977. Southern California trawl-caught juvenile rockfishes (Preliminary). S. Calif. Coastal Water Res. Proj., El Segundo, CA. Proc. Taxonomic Standardization Program 5 (4): 25-32.

Allen, M.J. In prep. Annotated checklist of marine and freshwater fishes of California.

Cooper, L.D. In prep. Guide to gross external diseases and anomalies of trawl-caught fishes and invertebrates of southern California.

Eschmeyer, W. N., E. S. Herald, and H. Hammann. 1983. A field guide to Pacific Coast fishes of North America. Houghton Mifflin Co., Boston, MA. 336 pp.

Kramer, D. E., W. H. Barss, B. C. Paust, and B. E. Bracken. 1995. Guide to northeast Pacific flatfishes. University of Alaska, Fairbanks, Alaska Sea Grant College Program, Fairbanks, AK. 104 pp.

Mearns, A. J., and M. J. Allen. 1978. Use of small otter trawls in coastal biological surveys. U. S. Environ. Prot. Agcy., Environ. Res. Lab., Corvallis, OR. EPA-600/3-78-083. 33 pp.

Mearns, A. J., and P. L. Haaker. 1973. Identifying and coding color anomalies in flatfishes. S. Calif. Coastal Water Res. Proj., El Segundo, CA. TM 200. 6 pp.

Miller, D. J., and R. N. Lea. 1972. Guide to the coastal marine fishes of California. Calif. Dep. Fish Game, Fish. Bull. 157. 249 pp.(addendum in 1976).

Robins, C. R., R. M. Bailey, C. E. Bond, J. R. Brooker, E. A. Lachner, R. N. Lea, and W. B. Scott. 1991. Common and scientific names of fishes from the United States and Canada. 5th edition. Am. Fish. Soc., Spec. Publ. 20. 183 pp.

SCAMIT. See Southern California Association of Marine Invertebrate Taxonomists.

Southern California Association of Marine Invertebrate Taxonomists. 1998. A taxonomic listing of soft bottom macro and megainvertebrates from infaunal and epibenthic monitoring programs in the Southern California Bight. Edition 3. S. Calif. Assoc. Mar. Invert. Tax., San Pedro, CA. 168 pp.

Stubbs, H. H., D. W. Diehl, and G. P. Hershelman. 1987. A Van Veen grab sampling method. S. Calif. Coastal Water Res. Proj., Long Beach, CA. Tech. Rep. 276. 4 pp.

Tetra Tech. 1986. Quality Assurance and Quality Control for 301(h) Monitoring Programs: Guidance on field and laboratory methods. Final report prepared for U. S. Environ. Prot. Agency, MOD/OMEP. Contract No. 68-01-6938.

Appendix 1:  Bight'98 Station location charts
Appendix 2:  Bight'98 Field Sampling - Sampling Organizations
Appendix 3:  Bight'98 Sample Processing Analytical Laboratories
Appendix 4:  Equipment and Supply lists
Appendix 5:  Sampling Vessel Specifications
Appendix 6 Data Sheets:
    Cruise Log
    Benthic data sheet
    Trawl data sheet
    Trawl fish species
    Trawl fish size class
    Trawl invertebrate species
    Trawl debris data
    Chain of custody form
    Sample tracking FAX transmittal sheet
    Bioaccumulation fish tracking sheet
Appendix 7 QA/QC Data Sheets:
    Trawl equipment
    Trawl processing checklist: part 1
    Trawl processing checklist: part 2
    Trawl processing checklist: part 3
Appendix 8:  Organization Contacts
Appendix 9:  SCCWRP Sample Shipping Information