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EPA/620/R-01/003

June 2001

National Coastal Assessment

Field Operations Manual

by

Charles J. Strobel

U. S. Environmental Protection Agency

Office of Research and Development

National Health and Environmental Effects Research Laboratory

Atlantic Ecology Division

Narragansett, RI 02882

and

Tom Heitmuller

U.S. Geological Survey

National Wetlands Research Center

Gulf Breeze Project Office

Gulf Breeze, FL 32561

DW14938557

Project Officer

J. Kevin Summers

Gulf Ecology Division

National Health and Environmental Effects Research Laboratory

Gulf Breeze, FL 32561
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NOTICE

The U.S. Environmental Protection Agency through its Office of Research and

Development collaborated in the research described here under interagency agreement

#DW14938557 to U. S. Geological Survey, National Wetlands Research Center. It has been

subjected to Agency review and approved for publication. Mention of trade names does not

constitute endorsement or recommendation for use.

This field manual represents the evolution of past EMAP-Virginian Province

manuals. As such, much of this document was left as originally written for the Coastal 2000

program. As a result, the reader will encounter the terms “Coastal 2000, C2000, CM,” etc.;

in most cases those terms now imply National Coastal Assessment (NCA). I would also like

to acknowledge the contribution past “EMAPers” have made to those documents.

Contributors have included Dan Reifsteck, Ray Valente, Jill Schoenherr, Darryl Keith, Steve

Schimmel, Kellie Merrell, Rebecca Fischman, Mike Daly, Don Cobb, and Kelly Byron.

The appropriate citation for this report is:

 U.S. EPA. 2001. National Coastal Assessment: Field Operations Manual. U. S. Environmental Protection Agency, Office of Research and Development, National Health and

Environmental Effects Research Laboratory, Gulf Ecology Division, Gulf Breeze, FL.

EPA 620/R-01/003. pp72.
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FOREWORD

The core elements of this field manual were prepared by Charles Strobel, EPA - Atlantic

Ecology Division, as a guidance document for the state teams responsible for field collections of

samples and data for EMAP’s Coastal 2000 Monitoring in the northeastern (NE) region of the

U.S. (Delaware to Maine). Although originally written specifically for NE field teams, the

methods described in the manual reflect standard procedures developed by EMAP-Estuaries

during 10 years of coastal monitoring activities conducted in all coastal regions of the contiguous

U.S. This manual has been modified to reflect a more national focus.

The overall objective is to put a practical field methods guide into the hands of the

participating field teams that allows a reasonable degree of flexibility to individual states, while

at the same time, provides adequate structure to ensure the uniform collection of comparable

field data on a national basis. This manual is intended as a “living document;” additional

sections can, and should, be appended as needed.
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CONTENTS

SECTION .................................................................................................................. PAGE

1 - Introduction ................................................................................................................... 1

2 - Overview of Field Sampling Activities ......................................................................... 3

Sampling Period ..................................................................................................... 3

Sampling Design..................................................................................................... 3

Indicators of Ecosystem Health .............................................................................. 3

Site Reconnaissance ............................................................................................... 5

Station Location ...................................................................................................... 5

Sampling ................................................................................................................. 5

3 - Field Data Base Management........................................................................................ 7

Sample Tracking Procedures .................................................................................. 7

Station and Sample Numbers ................................................................................. 7

Use of Bar Codes .................................................................................................... 8

Electronic Data Entry ............................................................................................. 8

4 - Water Quality Measurements ...................................................................................... 12

Hydrolab H20/Scout ............................................................................................. 12

Light Attenuation .................................................................................................. 16

Secchi Depth ......................................................................................................... 17

5 - Water Column Nutrients.............................................................................................. 18

Chlorophyll a and Phaeophytin ............................................................................ 18

Dissolved Nutrients .............................................................................................. 19

Total Suspended Solids......................................................................................... 20

Quality Control ..................................................................................................... 20

6 - Sediment Collections .................................................................................................. 21

Sediment Collections ............................................................................................ 21

Field Processing of Samples for Benthic Community Assessment ...................... 22

Field Processing of Samples for Chemistry and Toxicity Testing ........................ 25

Quality Control/Quality Assurance ...................................................................... 26

Safety Considerations ........................................................................................... 28
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SECTION .................................................................................................................. PAGE

7 - Fish Trawls .................................................................................................................. 29

Gear and General Protocols .................................................................................. 29

Trawl Preparation ................................................................................................. 30

Net Deployment .................................................................................................... 31

Trawling ................................................................................................................ 31

Net Retrieval ......................................................................................................... 33

Safety Considerations ........................................................................................... 33

Criteria for Voiding Tows ..................................................................................... 34

Endangered Species .............................................................................................. 34

Sample Processing ................................................................................................ 34

Quality Assurance................................................................................................. 40

Contingency Plans ................................................................................................ 40

Collection Permits ................................................................................................ 41

8 - Packaging and Shipping Samples ............................................................................... 42

Proper Packaging Methods ................................................................................... 42

Benthic Biology Samples ..................................................................................... 44

Sediment Chemistry Samples............................................................................... 44

Sediment Toxicity Samples .................................................................................. 45

Grain Size Samples............................................................................................... 45

Chlorophyll, Nutrients, and Total Suspended Solids ............................................ 45

Fish Chemistry ...................................................................................................... 45

Pathology QA Samples ......................................................................................... 46

Instructions for FEDEX Shipping with Dry Ice ................................................... 46

Appendices

List of supplies and equipment ......................................................................................... 47

Trawl net specifications .................................................................................................... 52

Coastal 2000 data sheets ................................................................................................... 55

Water Quality Measurement by Hydrolab DataSonde 3 /Surveyor 4 ............................... 64
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SECTION 1

INTRODUCTION

As a regulatory agency, the U.S. Environmental Protection Agency (EPA) is charged with

the mission to set environmental policy, obtain funds for research and development, and evaluate

the efficacy of environmental regulations in preserving the Nation’s natural resources. EPA’s

National Coastal Assessment (NCA), also referred to as Coastal 2000, C2000, CM in this

document, is a five-year effort led by EPA’s Office of Research and Development to evaluate the

assessment methods it has developed to advance the science of ecosystem condition monitoring.

C2000 represents the current state of evolution of EPA’s Environmental Monitoring and

Assessment Program (EMAP). EMAP was originally designed to provide a quantitative

assessment of the regional extent of environmental problems by measuring status and change in

selected indicators of ecological condition. EMAP provides a strategy to identify and bound the

extent, magnitude, and location of environmental degradation and improvement on a regional

scale.

Beginning in the year 2000, C2000 will attempt to assess the condition of the Nation’s

estuarine waters through statistically valid subsampling. Whereas the original EMAP effort was

conducted primarily by EPA and contract staff, C2000 is being implemented in partnership with

the 24 coastal states. This partnership recognizes that each of these entities plays an important

role in estuarine monitoring. Wherever possible, existing state monitoring programs are being

incorporated into the C2000 design. This provides for the maximum utilization of a limited

budget, and the flexibility of allowing states to often maintain historical sampling designs. Many

of these state programs have been in existence for many years, providing a basis for possible

C2000 trends analyses. Each state will conduct the survey and assess the condition of their

coastal resources independently. These estimates will then be aggregated to assess the condition

at EPA Regional, biogeographical, and National levels. Through this partnership EPA hopes to

build infrastructure within the coastal states to improve, and make more inter-comparable, the

multitude of estuarine monitoring programs throughout the country.

As stated above, C2000 is being implemented in cooperation with the coastal states.

Most of the field sampling, and some of the sample analysis, will be conducted by state agencies

through cooperative agreements with EPA. Analyses which state agencies choose not to perform

will be conducted through a national-level contract with qualified laboratories. A common suite

of “core” indicators will be measured using comparable methods:

C sediment contaminant concentrations

C sediment toxicity (Ampelisca abdita)

C benthic species composition

C sediment characteristics (grain size, organic carbon content, percent water)

C water column dissolved nutrients

C chlorophyll a concentrations,

C total suspended solids concentration,
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C surface and bottom dissolved oxygen, salinity, temperature, and pH

C water clarity

C contaminant levels in fish and shellfish

C external pathological condition of fish

C fish community structure

C2000 is designed as multi-year program. In the northeast portion of the United States

(Delaware to Maine), estuarine waters will be sampled over a two-year span (2000-2001).

Approximately 30 to 40 stations will be sampled per state each year. Tentatively, the following

two or three years will be dedicated to other ecosystems, such as coastal waters and/or salt

marshes, with the hope of returning to estuaries in years five and six.

Each major region will be coordinated through a central EPA location:

EPA’s Atlantic Ecology Division (AED), Narragansett, RI - Northeast States

EPA’s Gulf Ecology Division (GED), Gulf Breeze, FL - Gulf and Southeast States and

Puerto Rico

EPA’s Western Ecology Division (WED), Newport, OR - West Coast, Alaska, and Hawaii

EPA’s Mid Ecology Division (MED), Duluth, MI - Great Lakes

The purpose of this manual is to document suggested field data and sample collection

procedures for C2000-NE. These protocols have been developed by EMAP over the past 10

years. They will be identical to, or at least comparable with, those used in other regions of the

country. Individual states may prefer to use other methods, especially if they are currently being

used in existing programs. This is acceptable providing that comparability can be demonstrated

to the appropriate Regional Field Coordinator and the QA Officer.
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SECTION 2

OVERVIEW OF FIELD SAMPLING ACTIVITIES

2.1 Sampling Period

The sampling period for C2000 is based on the historical index period established for

EMAP-Estuaries efforts. This is based on the time frame in which the benthic biota are most

active and hypoxia is most prevalent. The established index period is July through September.

Some deviation from this period may be acceptable if the criteria for defining the index period

are met.

2.2 Sampling Design

The EMAP-Estuaries sampling design on which C2000 is based combines the strengths

of systematic and random sampling with our understanding of estuarine systems. It provides a

design that will allow probability-based estimates of the status of the Nation’s estuarine systems,

the variability associated with that status, its spatial and temporal components, and the temporal

trends associated with changes in these systems. The Coastal 2000 sampling design is based on a

single, annual sampling season of each station during the Index Period. The design differs from

previous EMAP designs in that existing monitoring programs were incorporated where

appropriate. “Biased” programs, such as those designed to evaluate the effects of a treatment

plant, would NOT be appropriate for inclusion. Working with the states, the C2000 design team

was able to identify a large number of sites that are currently being monitored and meet the

criteria for being unbiased in their location. Many were randomly located in the original

monitoring design.

The objective of the sampling design is to provide a statistically defensible strategy for

collecting information about selected indicators of ecological condition and their variability. The

design is flexible to allow alternative future uses.

2.3 Indicators of Ecosystem Health

The primary goal of C2000 is to provide an assessment of overall ecosystem condition.

To accomplish this goal, a number of “indicators” of ecosystem health will be monitored. The

core set of indicators agreed upon by all entities involved in this project is listed in Table 1, with

the goal being to collect data on all of these indicators at all sampling stations.
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Table 1. List of core ecological indicators being measured by C2000

Water Quality Indicators

Hydrographic Profile

- dissolved oxygen

- salinity

- pH

- temperature

- depth

- light attenuation (PAR, transmittance)

- secchi depth

Water Quality Samples

- dissolved nutrients (ortho-phosphates, nitrites, nitrates, ammonia)

- chlorophyll a

- total suspended solids (TSS)

Sediment Quality

Composited Surficial Sediment

- sediment contaminants (organics and metals)

- sediment TOC

- sediment toxicity (amphipod)

- percent silt/clay

Biota

Fish/Shellfish

- community structure (species; abundance; total length, up to 30 individuals)

- tissue contaminants (organics and metals)

- external pathology (fish)

Benthos

- community structure (standard grab - 0.04m2

)

Habitat

- SAV (presence/absence)

- basic habitat type (e.g., open water, tidal flat, marina, harbor, inlet, tidal river/stream,

seagrass bed, sandy/muddy bottom, rocky bottom, shelly bottom, coral reef, etc.)

- marine debris (presence/absence)
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2.4 Site Reconnaissance

Prior to the start of field activities, a thorough reconnaissance of the area to be sampled

should be performed whenever field crews will be working in new areas. This includes

determining the locations of boat ramps, hotels, and dry ice suppliers; visiting any stations that

may fall in water too shallow for boats; and attempting to identify any potential problems that the

field crews may face during the Index Period. Needless to say, reconnaissance may not be

needed in areas with which state field crews are familiar.

2.5 Station Location

The randomly selected sampling locations for each state (or specific study area) will be

provided to the field crews as coordinates of latitude/longitude in degrees-minutes, expressed to

the nearest 0.01 minute (i.e., 00E 00.00'). The crews will use GPS (preferably DGPS) to locate

the site. Three different locations will be provided for each station. These are identified as “A”,

“B”, and “C”. The primary site is the “A” location; “B” and “C” are backups. If the primary site

is not accessible, or the bottom is too rocky to obtain a sediment sample, then the crew may

move to the “B” site. If that site is also unsampleable, then they should move to the “C” site. If

all three are unsampleable, then the site is not sampled. If one of the sites can be sampled for

only some of the indicators, then that sampling should be conducted. It is important that the

crew note on the datasheet at which of these locations (A, B, or C) the samples were collected.

Crews will attempt to navigate to the location to within 0.02 nm (± 37 meters) of the

given coordinates. This reflects the accuracy expected from a properly functioning GPS unit of

the caliber that will used for the study. The crew will record the actual coordinates of the vessel

after anchorage, NOT the initial intended coordinates, on the field data sheet.

2.6 Sampling

In order to collect data as efficiently as possible and reduce the potential for sample

contamination, the samples should be collected in the order shown in Figure 1. Details about

each sampling procedure can be found in following chapters. A complete list of needed supplies

and equipment can be found in Appendix A. As stated in the Introduction, variations from these

methods must be approved, in advance of sampling, by the Field Coordinator and QA Officer.

An overview of Quality Control/Quality Assurance (QC/QA) protocols for each sampling

technique can be found after the description of each procedure. A more detailed account of QC/

QA proposals can be found in the C2000 Quality Assurance Plan.
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SECTION 3

FIELD DATA BASE MANAGEMENT

Management of data in the field is of paramount importance. Without proper data

management the quality of the data generated is questionable. Field data management consists of

two categories; written data sheets and electronic data.

In general all data will be recorded on hard-copy datasheets while on-station, and entered

into a computer back on shore. The use of bar code readers will facilitate the entry of sample

numbers and eliminate transcription errors. EPA can provide datasheets and electronic forms for

data entry if desired. EPA can also assist in the procurement of bar codes. C2000 datasheets are

included in Appendix C. Although their use is not required, it is highly recommended.

It is the responsibility of the chief scientist to guarantee the quality of the data. At the end

of each day it is his/her responsibility to review the data collected that day and “sign-off” on it.

3.1 Sample Tracking Procedures

A variety of water, sediment, and biological samples and data are collected during the

C2000 sampling effort. These include physical samples (i.e., sediment and nutrient samples) and

non-physical samples (i.e., Hydrolab or YSI cast data). It is vital that all of these samples and

data be tracked from collection to the receipt of analytical results. To accomplish this purpose,

all samples collected are assigned unique sample identification numbers (SAMPLE IDs)

composed of the station number and a sample-type code. These numbers are used to track

samples from collection to inclusion in the final National database.

3.2 Station and Sample Numbers

Because Coastal 2000 is a large-scale national monitoring program being implemented by

many agencies with data feeding into a centralized database, it is critical that all stations and

samples be assigned unique identifiers. All information sent to the national database must be

associated with a station using the C2000 convention. Station names will include the state in

which the station is located, the year of sampling, and a number. The format adopted is as

follows:

SSYY-XXXX

Where,

SS is the state where the station is located,

YY is the last two digits of the year (00 for year 2000), and

XXXX is a four-digit incremental identifier, beginning at 0001.
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So, for example, the first 35 stations in Massachusetts, Texas, and California, sampled in year 2000

would be identified as:

MA00-0001 to MA00-0035

TX00-0001 to TX00-0035

CA00-0001 to CA00-0035

Note that where one state entity will be sampling in another state’s waters, the station number is

based on the “location” not the organization conducting the sampling. This is likely to occur

when one state will be responsible for an entire water body even though portions fall within the

neighboring state’s jurisdiction.

Sample numbers will be made up of the station code with a sample type identifier

attached to the end. Sample number formats are illustrated in Table 2. All sample information

sent to the national database must use this format.

3.3 Use of Bar Codes

The use of bar codes to label samples is highly recommended. Ten years of experience

with EMAP has demonstrated their utility. Bar codes are preprinted for every sample that might

be collected during the sampling season, with side-by-side duplicates for each sample, by a

professional service. Each label contains both the bar code itself, and the printed sample number.

Labels are waterproof and do not come off when frozen or immersed in formalin.

When a sample is collected, one of the duplicate labels is placed on the sample and the

other on the datasheet. When the data are transcribed from the datasheet into the computer, the

sample numbers need not be typed in. They can just be scanned with a bar code reader. This

virtually eliminates transcription error.

Bar codes also make shipping of samples easier. As a sample is placed in the shipping

container, the sample ID is scanned into the computer. A packing list can then be printed out for

inclusion with the shipment.

3.4 Electronic Data Entry

All information recorded on the datasheets must be entered into a computer for eventual

transfer to the national database. This should be done as soon as possible, while the sampling

event is “fresh” in the crew’s mind. It is the responsibility of the chief scientist to ensure the

accuracy of the electronic data file.

One method to facilitate this process is to use form-filler software. This software can be

used to design and print hard-copy datasheets, and to create an identical electronic form. Having

the computer screen as an exact copy of the field forms facilitates data entry. QA is easier as the

completed electronic form can be printed out for side-by-side comparison with the original field

form.
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EPA can provide both the field and electronic datasheets to any participant desiring them.

The participant would only need to purchase the appropriate commercial software to access the

electronic forms.
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Table 2. Sample numbers assigned to each sample type. Sample number consists of the station number

(state/year - number, e.g., RI00-0001 for Rhode Island, year 2000, station 1) followed by a sample

type code. The list below uses station RI00-0001 as an example to illustrate the appropriate sample

coding. For QA samples, the state identifier is replaced with “QA” and the station designator is

sequential rather than being associated with a given station. The link is made in the database. While

collection of specific QA samples is helpful, EPA does not require all participants to collect these

samples.

Sample Type Type Code Example Sample Number Bar-Coded?

CTD Cast CTD RI00-0001-CTD N

Light measurement (PAR) profile PAR RI00-0001-PAR N

Surface chlorophyll SCL RI00-0001-SCL Y

Surface suspended solids SSS RI00-0001-SSS Y

Surface dissolved nutrients SN RI00-0001-SN Y

Mid-depth chlorophyll MCL RI00-0001-MCL Y

Mid-depth suspended solids MSS RI00-0001-MSS Y

Mid-depth dissolved nutrients MN RI00-0001-MN Y

Bottom chlorophyll BCL RI00-0001-BCL Y

Bottom suspended solids BSS RI00-0001-BSS Y

Bottom dissolved nutrients BN RI00-0001-BN Y

Benthic infauna(1) BI1 RI00-0001-BI1 Y

Benthic infauna(2) BI2 RI00-0001-BI2 Y

Benthic infauna(3) BI3 RI00-0001-BI3 Y

Sediment Toxicity ST RI00-0001-ST Y

Sediment grain size SG RI00-0001-SG Y

Sediment Organics SO RI00-0001-SO Y

Sediment Metals SM RI00-0001-SM Y

Sediment TOC OC RI00-0001-OC Y

Standard fish trawl STRL RI00-0001-STRL N

Non-standard fish trawl TRL RI00-0001-TRL N

Fish chem. species 1 composite FC1 RI00-0001-FC1 Y

Fish chem. species 1 individuals FC1-1 to FC1-9 RI00-0001-FC1-1

To RI00-0001-FC1-9 Y

Fish chem. species 2 composite FC2 RI00-0001-FC2 Y

Fish chem. species 2 individuals FC2-1 to FC2-9 RI00-0001-FC2-1

To RI00-0001-FC2-9 Y

“Other” sample type 1 OTH-1 RI00-0001-OTH-1 Y

“Other” sample type 2 OTH-2 RI00-0001-OTH-2 Y

Quality Assurance Samples (full range of sample numbers given for C2000-Northeast)

Fish pathology QA PATH QA00-0001-PATH

To QA00-0500-PATH Y

Chlorophyll QA CL QA00-0001-CL

To QA00-0300-CL Y

Dissolved nutrients QA N QA00-0001-N

To QA00-0300-N Y

TSS QA SS QA00-0001-SS

To QA00-0300-SS Y
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SECTION 4

WATER QUALITY MEASUREMENTS

One of the activities to be performed at every station is the collection of water quality

information (salinity, temperature, pH, and dissolved oxygen [DO]). At each station a vertical

profile through the water column will be obtained using a profiling instrument. Multiparameter

water quality probes are available from several manufacturers (e.g., Hydrolab®, Yellow Springs

Instruments, YSI®, and SeaBird®); each manufacturer offers various models with varying

capabilities. The following instructions are specific for crews using the Hydrolab H20 probe in

conjunction with the Scout 2 deck display; these procedures will require some modification when

different, but similar instrumentation is used. Appendix D details the calibration and use of the

Hydrolab DataSonde3/Surveyor 4, the array used extensively by NE crews which requires the use

of a PC and the software package PROCOMM to setup or calibrate.

4.1 Hydrolab® H20/Scout 2 - Hydrographic Profiling Instrumentation

The Hydrolab H20 probe/stirrer used with the Scout 2 Display Unit is powered by an

internal battery pack of 10 conventional, alkaline “AA” batteries . This combination provides a

self-contained, hand-held water quality data system that displays real-time measurements. The

terms “Hydrolab”, “sonde”, or “unit” may be used instead of H20/Scout 2 when referring to the

instrumentation.

4.1.1 Setup and Calibration

The following is a brief description of the calibration procedures for the Hydrolab H20/

Scout 2 pairing. Refer to the manufacturer’s manual for detailed discussion of the equipment,

including maintenance and repair information. Calibration should be performed every morning

prior to the start of the day’s sampling and documented in the “QC CHECK” block appearing

near the top of the Hydrographic Profile field data form (see attached form). It is recommended

that a “field calibration kit” consisting of the following supplies and items be assembled and

stored in a good quality, plastic tackle box:

- calibration cup with removable cover

- DO membranes

- backup probes and spare parts (e.g., pH and DO probes, o-rings, etc.)

- standard seawater (known salinity, 15-22 ppt)

- pH buffers (7 and 10)

- laboratory thermometer

- deionized (DI) water

- ring stand with large clamp

- squeeze bottle (rinsing)

- calibration forms

- appropriate size leak-proof, screw-cap containers (e.g., Nalgene) for the additional calibration

solutions

- basic tools (assorted screwdrivers, allen wrenches)
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A properly maintained Hydrolab that is used on a regular basis, should remain fairly stable and

free from marked drift or variation in its measurements from one day to the next. Daily QC checks of

the Hydrolab provide the user with immediate feedback on the instrument’s general reliability and a

written performance record for each of the parameters. Based on years of experience with Hydrolab

units, EMAP considers the following ranges acceptable for day-to-day instrument drift:

DO ± 0.5mg/l (or ± 8% saturation)

pH ± 0.3 units

salinity ± 1.0 ppt

temperature ± 1.0EC

depth ± 0.5m

These acceptability ranges take into consideration real-world factors to which a unit is typically

exposed during the rigors of a day in the field (e.g., pounding seas, temperature extremes, rough

rides in a truck, etc.). Usually, a well maintained Hydrolab will remain within tighter agreement

than the ranges listed. Although the above criteria represent acceptable instrument drift, on the

morning of each field day, the unit still should be calibrated to correct for drift as much as

possible (see QAPP for additional discussion).

Performance that falls outside of the acceptability range or sluggish equilibration

response, signals that the unit needs maintenance focused on the deficient functions (e.g., battery

replacement, DO membrane replacement, pH reference solution renewal, cleaning electrodes,

etc.). After determining and correcting the problem, the Hydrolab should readily accept new

calibration values and quickly equilibrate. If performance remains substandard, the unit should

be removed from service until it is repaired. It is highly advisable to have a backup unit available.

1. General Setup

Attach the probe to the deck unit using data cable; make sure the connections are locked

or secured. Power up the unit by pressing the on/off switch; display window should illuminate;

bring up primary operating screen. Remove the storage cup from the probes and replace with the

calibration cup. Hold or use ring-stand clamp to secure the H20 in an inverted (probes up)

position during calibration procedures. The preferred sequence of calibration is temperature,

salinity, pH, DO, and depth; regardless, salinity should be calibrated prior to DO.

2. Salinity Calibration and Temperature Check

Rinse probes with about 10 mls of salinity standard; dump and refill the calibration cup

with salinity standard to a level that completely covers the white conductivity cell. Place

thermometer into the calibration solution and allow to equilibrate; record the thermometer

reading to the nearest whole degree in the “STANDARD” box under the TEMP column; record

the temperature value appearing on the deck display as the “MEASURED” value for TEMP; the
14

measured temperature should be within ±1EC of the standard. NOTE: the temperature function for the

H20 is set at the factory and cannot be corrected in the field; it must be sent back to Hydrolab for

adjustments. There is no field calibration procedure for temperature, but rather a QC check to verify

proper function.

Continue with the salinity cal check. First record the “true” value of the salinity standard in the

STANDARD box under SAL; then, record the salinity reading from the display screen in the

MEASURED box. If MEASURED differs from STANDARD, calibrate the function.

To calibrate (see Hydrolab Operating Manual for detailed discussion): on the deck display,

depress the “Calibrate” key to bring up the menu of variables (abbreviations or symbols); scroll

across the menu by keying the right or left direction arrows to “S” for salinity; initiate the

calibration. The screen will display the sensors current reading; use directional (R/L) arrows to

scroll to the numerical character that needs to be adjusted, then use the up/down arrows to

increase or decrease the value to represent the true value of the standard. Press “‘Enter” to

calibrate, then select Y (yes) to accept the new calibration value for salinity. Return to Main

Screen to verify the calibration; the displayed value for salinity should be within ± 0.5 ppt of the

standard.

Carefully pour the salinity standard solution from the calibration cup back into the field supply

bottle. It is permissible to reuse the salinity standard for several days as long as precautions are

taken to ensure that the solution is not diluted or allowed to evaporate. Leave the calibration cup

on the instrument and rinse the probes and cup with a few squirts of DI water; dump and shake to

remove residual, then proceed to the pH calibration.

3. pH Calibration

Two buffer solutions are required to set up the pH calibration; pH 7 and pH 10 standards

are recommended. First, pour pH 7 buffer into calibration cup to a level that completely covers

the reference pH cannister. Bring up Main Screen and allow the pH reading to equilibrate

(approximately 20-30 secs); record the pH reading on the QC Check form in the MEASURED

box under pH 7; then proceed to calibrate in similar manner as described for salinity, only select

the pH variable from menu. Key in Calibration mode; scroll with arrows; adjust value to standard

(7.00); enter new value and accept by selecting “y” (yes); record 7.0 in the CALIBRATE box.

Decant pH 7 buffer and rinse probes with DI water. Repeat process with pH 10 to complete the

two-point calibration slope.

4. Dissolved Oxygen (DO) Calibration

Remove the plastic guard from the DO probe to fully expose the membrane and fill the

calibration cup with DI water to a level just to, but not over the DO membrane (even with the Oring); gently wipe any water droplets from the membrane using a tissue. Place the cap over the

calibration cup, then allow 2-3 minutes for the environment around the DO membrane to become

saturated with moisture (this system of calibration is commonly referred to as water saturated-air
15

calibration). After the system has equilibrated, switch to Alternate Screen to the display that reads out

“% Saturation.” The standard value is 100%; record the instrument’s measured value in the

MEASURED box on QC Check form. Any value $ 95% is good indication that the DO function is

performing well. To calibrate DO, press the “Calibration” key; scroll over to “%” and select; a screen

will appear with that displays standard pressure at 760 mm (normal for sea level); press the Enter key

and Y to accept calibration. Return to Alternate Screen to verify that DO calibration was successful -

100 ± 2%. Record the instrument reading in the CALIBRATE box.

5 Depth Sensor

The depth sensor on the H20 is a transducer that responds to water pressure. For the daily

QC Check on depth, when the unit is out of the water, the transducer experiences no significant

pressure and the depth sensor should register 0.0 meters; therefore, on the QC Check block, the

standard value for depth is 0.0 m. For QC Check, refer to the main screen and record the

displayed depth measurement in the MEASURED box. If the reading is not 0.0, calibrate by

pressing the “Calibration” key, then scroll over to “D” and press Enter. Adjust the depth reading

to 0.0 by using the up/down arrows; Enter the new calibration value and accept by selecting

“Y”(yes) key followed again with Enter. Record the instrument’s depth reading in the

CALIBRATE box.

At the conclusion of calibration, be sure to turn off the power (to conserve battery life).

Remove the calibration cup and replace with the storage cup. Be sure that it contains a small

amount of water (approximately 100-200 ml); the probes must be maintained in a wet

environment.

Field Verification for Depth

To further verify the accuracy of the depth function, while in the field, lower the sonde to

a known depth (e.g., 2 meters via marked data cable) and compare to the instrument’s reading for

depth. If noticeably different (e.g., 0.2 m), re-calibrate by performing the calibration procedures

above, except use the “known” depth as the STANDARD.

NOTE: When conducting water column profiles, the preferred determination for depth is

to use an accurately marked cable line. However, in conditions of strong tides and currents, due

to the scope on the cable, it may be necessary to utilize the depth sensor to estimate depth.

4.1.2 Field Deployment and Water Column Measurements

After successfully completing the daily QC Checks/Calibration, the Hydrolab is ready for

use. A five gallon bucket is a convenient storage/transport container.

Once aboard the vessel, the unit can be readied for use by removing the protective cup

and attaching the stirrer; however, the exposed probes must be held in water. The bucket can be

filled to a level that covers the stirrer and probes, or, a storage well can be constructed from 6-

inch PVC piping. Cap one end of a 24-in section of pipe with a plug and affix the resultant well
16

to the vessel railing (or other suitable point of attachment) with line or tape. Insert the sonde into the well

and fill with site water; the instrument can then remain intact and ready to go for the duration of a

sampling day.

A station of nominal depth (<10 m) field measurements for the water column profile will

be taken at near surface (probes approximately 0.5 m below surface), at 1-meter intervals down

the water column, and at 0.5 m off bottom. On the down cast, at each interval, the parameters of

interest are recorded on the Hydrographic Profile data form; a replicate set of measurements is

recorded at each interval coming back up the water column. See QAPP for discussion on

sampling interval for deep stations (>10 m).

Remember to always turn the power off after completing the water column profile; a unit

left on with the stirrer attached will quickly drain the batteries.

4.2 Light Attenuation

C2000-NE crews will also obtain a vertical profile of light for the purpose of calculation

of the light attenuation coefficient at each station. This can be accomplished using either a PAR

(photosynthetically active radiation) meter or a transmissometer. This profile can be obtained in

conjunction with the CTD profile or separately, depending upon the equipment available. PAR

sensors require no field calibration, however, they should be returned to the manufacturer prior to

each field season for annual calibration.

To obtain a PAR profile using an independent datalogger such as the LI-COR LI-1400:

1. Connect a deck sensor and an underwater sensor to the LI-1400. Make sure the correct

calibration factors are entered for each probe. These are supplied by the manufacturer.

2. Place the deck sensor on the boat in a location where it will is not shaded.

3. Lower the underwater sensor on the SUNNY (or at least unshaded) side of the boat to a depth

of about 10 cm (represents “surface”).

4. Once readings stabilize, record the values from both sensors (FE/m2

/s), along with the water

depth of the underwater sensor, on the datasheet. Log the values in the datalogger.

5. Lower the underwater sensor to 0.5 meters, allow the values to stabilize, and record the

values from both sensors, along with the water depth of the underwater surface.

6. Repeat at the following schedule:

Shallow sites (# 2 m) - every 0.5 m interval;

Nominal depths (>2<10 m) - 0.5 m (near-surface) and every 1-m interval to near-bottom (0.5

m off-bottom);
17

Deep sites (>10 m) - 0.5 m (near-surface) and every 1-m interval to 10 m, then at 5-m

intervals, thereafter, to near-bottom (0.5 m off-bottom).

7. If the bottom is impacted with the meter, allow 2-3 minutes for the disturbed conditions to

settle before taking the reading.

8. If the light measurements become negative before reaching the bottom, terminate the profile

at that depth.

9. Repeat the process on the upcast.

4.3 Secchi Depth

The Secchi disk is used to give a measurement of the transparency of the water column,

also called the secchi depth. This measurement is made at every station and is recorded on the

CTD datasheet. A 20 cm black and white Secchi disk is held by a non-stretch line that is marked

in two tenths of a meter intervals. To determine the Secchi depth:

1. Slowly lower the Secchi disk on the shady side of the boat until it is no longer visible and

note the depth using the markings on the line (interpolate between markings to the nearest 0.1

meter). If the disk hits the bottom, meaning the Secchi depth is greater than the water depth,

note this on the datasheet.

2. Slowly raise the Secchi disk until it just becomes visible and note the depth.

3. Perform steps 1 and 2 three times, noting both readings. Record the average of the readings.

QUALITY CONTROL FOR SECCHI DISK

1. If the range of measurements for the three sets of depth readings is greater than 0.5 m, the

entire process should be performed again.

2. No sunglasses or any other devices should be used to shade the eyes while this procedure is

being performed.

3. The Secchi depth should be determined from the shady side of the boat during daylight hours.
18

SECTION 5

WATER COLUMN NUTRIENTS

Water samples will be collected at each site and analyzed for:

Chlorophyll a

Dissolved ammonia, nitrites, nitrates, orthophosphates, and

Total Suspended Solids.

Samples should be collected at three depths; surface, mid-water, and bottom, depending

upon the depth of the water:

Shallow sites (< 2 m) - 0.5 m (near-surface) and 0.5 m off-bottom1

Standard sites (>2 m) - 0.5 m (near-surface), mid-depth, and 0.5 m off-bottom;

1

 Unless he depth is so shallow that near-surface and near-bottom overlap; then sample at mid-depth only.

Water samples should be obtained, either using a pumped system or a water sampling

bottle such as a Niskin or 5 Go-Flo® bottle, and transferred to a rinsed (3x with water from the

sampling bottle) one gallon HDPE container.

5.1 Chlorophyll a and Phaeophytin

Chlorophyll samples must be filtered no more than 4 hours after collection. Any further

delay is strongly discouraged due to the possible lysis of phytoplankton cells. Samples that

cannot be filtered immediately after collection must be held at 4EC until filtered. Filtering can be

accomplished by either of two methods. The first requires the use of a vacuum pump, either

electric or hand operated. The second uses positive pressure. The method used must be noted on

the datasheet.

5.1.1 Vacuum filtration

Immediately concentrate the algae by filtering onto two, separate 47 mm GF/F filter

pads; Process a sufficient amount of sample (i.e. 100-1,500 ml) to produce a green color on the

filter; the volumes of replicate samples should be the same. Record the volume filtered on the

datasheet. The filtrate should be saved for dissolved nutrient analyses (Section 4.2). To avoid cell

damage and loss of contents during filtering, do not exceed a vacuum of 15 psi or a filtration

duration of greater than 5 minutes. Add 1 ml of saturated MgCO3

 solution (10 mg/L) during the

last few seconds of filtering AFTER THE NUTRIENT FILTRATE HAS BEEN REMOVED.

This buffers the sample to reduce the possibility of degradation. Carefully remove the filters

using forceps (never touch the filter with your fingers), fold in half, and wrap in clean aluminum

foil or small disposable petri dish. Mark both the volume filtered and the sample number (SCL,
19

MCL, BCL: surface, mid, or bottom chlorophyll) on the foil envelope or petri dish. Place both filters in

a whirl pak or small disposable petri dish and affix the appropriate bar code or hand-write the sample

number. Place the package on dry ice.

Note that filter funnels should be rinsed with DI water prior to filtration. In addition,

graduated cylinders should be rinsed with site water.

If too much sample is filtered (i.e., there is a thick layer of material on the filter pad), it

should be discarded and the filtration repeated with a smaller volume. Too much material may

result in some “oozing out” when the pad is folded.

5.1.2 Positive pressure filtration

The alternative method is to use positive pressure to push a sample through the filter. A

disposable, graduated 50-cc polypropylene syringe fitted with a stainless steel or polypropylene

filtering assembly is used to filter the site water through 25 mm GF/F filters; the volume of water

filtered must be documented. If conditions allow (based on the suspended solids load), up to 200

ml of site water should be filtered for each chlorophyll sample; for a 50-cc syringe, that equates

to 4 fills. To refill when the syringe barrel empties, carefully detach the filter assembly and fill

the syringe to the mark, replace the filter and continue with the filtration until the desired volume

has been processed. The filtrate from this process is saved for the analysis of dissolved nutrients

(see Section 4.2). After filtering the sample, add 1 ml of saturated MgCO3

 solution (10 mg/L) to

the syringe (AFTER THE NUTRIENT FILTRATE HAS BEEN REMOVED) and pass this

through the filter pad. This buffers the sample to reduce the possibility of degradation. Using

tweezers, carefully remove the filter from its holder and fold once on the pigment side, then place

it in foil as described in 4.1.1. Record the volume of water filtered on both the foil and on the

field form. Mark the sample number on the foil pack, then place the foil pack in a whirlpack and

label the whirlpack with the appropriate bar code. Place on dry ice. Repeat the filtering process

for the second sample and store filter in the same whirlpack containing the first sample. The

samples must remain frozen until time of analysis. Discard the used syringe. Rinse the filtering

assembly with deionized water and store in a clean compartment between sampling stations (a

small tackle box makes a good carrying kit for supplies and equipment used in this activity).

5.2 Dissolved Nutrients

Approximately 40 ml of filtrate from the above chlorophyll filtration will be collected

into a prelabeled, clean 60-ml Nalgene screw-capped bottle and stored on dry ice. Before placing

sample in the freezer, affix the appropriate bar code to the bottle and record the approximate

salinity (±2 ppt) on the container. This is a convenience for the analyst who will perform the

nutrient analysis. Depending on the analytical instrumentation used, matrix matching of

solutions (e.g., standards or wash solutions) may be required for certain of the analytes. The

salinity value can be obtained from the water column data or by refractometer reading of the

actual water sample taken by sampling bottle. The nutrient samples should remain frozen until

time of analysis.
20

5.3 Total Suspended Solids

Approximately 1 liter of unfiltered seawater from the sampling bottle is poured into a 1-L

polypropylene bottle and stored at 4EC to await laboratory analysis.

5.4 Quality Control

Field duplicates: A field duplicate is a sample taken at the same location and depth as a

regular sample and processed for chlorophyll, nutrients, and total suspended solids. The

duplicate and sample should be taken in quick succession. A field duplicate should be collected

once for every 10 samples. The data from field duplicates indicates sampling precision.

Although some filtering may be done on shore, many times it will be necessary to filter

while on the boat. Working with liquids on a rocking boat presents many opportunities for

contamination, and therefore, special care must be taken. The following guidelines will help

prevent accidents while working with the water samples:

1. After every station empty the overflow bottle and all reservoirs.

2. Rinse the filtering apparatus with DI water before putting in a new filter.

3. Only handle filters with tweezers.

4. All filters should be inspected and damaged filters should be discarded.
21

SECTION 6

SEDIMENT COLLECTIONS

6.1 Sediment Collections

Sediments are collected for a variety of analyses. One grab sample is collected for

benthic species composition and abundance; the entire content of the grab will be sieved to

collect benthos. Additional sediment grabs are collected for chemical analyses, grain size

determination, and for use in acute toxicity tests. For these analyses, only surficial sediment (top

2-3 cm) will be collected. The surficial sediment from multiple grabs will be composited to

obtain an adequate volume (approximately 4-5 liters) of sample for the analyses; the number of

grabs needed may vary based on the sediment characteristics. While a biology grab is being

processed (sieved), grabs should be collected for chemistry/toxicity.

A 1/25 (0.04) m2, stainless steel, Young-modified Van Veen Grab sampler is used to

collect sediments. The sampler is constructed entirely of stainless steel and has been Kynar®-

coated (similar to Teflon) and is therefore appropriate for collecting sediment samples for both

biological and chemical analyses. The top of the sampler is hinged so the top layer of sediment

can be easily removed for chemical and toxicity analyses. This gear is relatively easy to operate

and requires little specialized training.

Other gear is also acceptable, following approval by the C2000 Field Coordinator. The

gear size must be identified on the appropriate datasheet.

Listed below is the protocol for obtaining sediment samples.

1. The sampler must be thoroughly washed with Alconox prior to use at a station, then

rinsed with ambient seawater to ensure no sediments remain from the previous station.

2. Attach the sampler to the end of the winch cable with a shackle and tighten the pin.

Attach a pinger to the grab.

3. Cock the grab.

4. Lower the grab sampler through the water column such that travel through the last 5

meters is no faster than about 1 m/sec. This minimizes the effects of bow wave

disturbance to surficial sediments.

5. Retrieve the sampler and lower it into its cradle on-board. Open the hinged top and

determine whether the sample is successful or not. A successful grab is one having

relatively level, intact sediment over the entire area of the grab, and a sediment depth at

the center of at least 7 centimeters (see Figure 2). Grabs containing no sediments,

partially filled grabs, or grabs with shelly substrates or grossly slumped surfaces are
22

unacceptable. Grabs completely filled to the top, where the sediment is in direct contact with

the hinged top, are also unacceptable. It may take several attempts using different amounts of

weight to obtain the first acceptable sample. The more weight added, the deeper the bite of the

grab. In very soft mud, pads may be needed to prevent the sampler from sinking in the mud. If

pads are used, the rate of descent near the bottom should be slowed even further to reduce the

bow wave.

6. Carefully drain overlying water from the grab. If the grab is used for benthic community

analysis, the water must be drained into the container that will receive the sediment to

ensure no organisms are lost.

7. Enter notes on the condition of the sample (smell, texture, presence of organisms on the

surface, etc.) on the benthic infauna data sheet.

8. Process the grab sample for either benthic community analysis or chemistry/toxicity

testing as described in Figure 3 and in Sections 6.2 and 6.3.

9. Repeat steps 4-8 until all samples are collected. To minimize the chance of sampling the

exact same location twice, the boat engines can be turned periodically to change the drift

of the boat, or additional anchor line can be let out.

6.2 Field Processing of Samples for Benthic Community Assessment

Grab samples to be used in the assessment of macrobenthic communities are processed in

the following manner:

1. Assign a sample number to the sample; affix the bar coded labels to the sample jar and

datasheet.

2. Measure the depth of the sediment at the middle of the sampler and record the value on

the data sheet. The depth should be >7 cm. Record descriptive information about the

grab, such as the presence or absence of a surface floc, color and smell of surface

sediments, and visible fauna in the computer.

3. Dump the sediment into a basin and then into a 0.5 mm mesh sieve. Place the sieve into

a table (sieve box) containing water from the sampling station. Agitate the tray in the

sieve box thus washing away sediments and leaving organisms, detritus, sand particles,

and pebbles larger than 0.5 mm. This method minimizes mechanical damage to fauna

that is common when forceful jets of water are used to break up sediments. A gentle flow

of water over the sample is acceptable. Extreme care must be taken to assure that no

sample is lost over the side of the sieve.
23
24

4. Drain the water from the sieve box and gently rinse the contents of the tray to one edge. Using

either your fingers or a spoon, GENTLY scoop up the bulk of the sample and place it in the

plastic screw-top bottle labeled in Step 1 (which should be placed in the sieve or a bucket in

case some of the sample spills over). Rinse the outside of the sample jar into the sieve, then,

using a funnel, rinse the contents into the jar. The jar should be filled no higher than the 700 ml

mark. If the quantity of sample exceeds 700 ml, place the remainder of the sample in a second,

unlabeled container. Using a waterproof marker, write the sample number on the second

container and tape the two together. Note on the datasheet that the sample consists of more

than one container.

5. Carefully inspect the sieve to ensure that all organisms are removed. Use fine forceps (if

necessary) to transfer fauna from the sieve to the bottle containing the proper sample

number.

6. Ten percent buffered formalin is used to fix and preserve samples. A 100 % buffered,

stained stock formalin solution should be mixed according to the recipe in Table 3. 100

ml of the formalin should be added to each sample jar, and a teaspoon-full of borax added

to assure saturation of the buffer. FILL THE JAR TO THE RIM WITH SEAWATER TO

ELIMINATE ANY AIR SPACE. This eliminates the problem of organisms sticking to

the cap because of sloshing during shipment. Gently invert the bottle to mix the contents

and place in the dark. If the sample occupies more than one container, tape all the sample

bottles containing material from that grab together.

7. Prior to sieving the next sample, use copious amounts of forceful water and a stiff brush

to clean the sieve, thereby minimizing cross-contamination of samples.

Table 3. Directions for mixing stock solutions of formalin.

Chemical Volume Desired Total Quantity

100% formalin stock (stained and buffered)

Rose Bengal stain 8 R 1/4 teaspoon

Borax 8 R 8 heaping tablespoons

100% formalin 8 R two gallons
25

6.3 Field Processing of Sediments for Chemistry and Toxicity Testing

In addition to the grab collected for benthic community analyses, additional grabs are

collected for chemical analyses and toxicity testing. The top 2 cm of these grabs are removed,

homogenized, and split for chemistry and toxicity testing. Because of contamination concerns

these samples are removed and processed in the order described below:

1. As each grab is retrieved, carefully examine it to determine acceptability. The grab

is considered acceptable as long as the surface layer is intact. The grab need not be

greater than 7 cm in depth for chemistry samples, but the other criteria illustrated in

Figure 2 apply. Carefully drain off, or siphon, any overlying water, and remove and

discard large, non-living surface items such as rocks or pieces of wood.

NOTE: Great care must be taken to avoid contamination of this sample from

atmospheric contaminants. The boat engine should be turned off or the boat

maneuvered to assure the exhaust is down wind.

2. A clean stainless steel or Teflon spoon is used to remove sediments from grab

samples for these analyses. All items must be washed with Alconox and rinsed

with ambient seawater before use.

3. Remove the top two cm of sediment using the stainless steel or Teflon spoon.

Place the sediment removed in a stainless pot and place the pot in a cooler on ice

(NOT dry ice). The sample must be stored at 4EC, NOT FROZEN.

4. Repeat this procedure, compositing the sediment in the same stainless pot until a

sufficient quantity of sediment has been collected for all samples (approximately 4

L). Stir sediment homogenate after every addition to the composite to ensure

adequate mixing. Keep the container covered and in the cooler between grabs.

5. Homogenize the sediment by stirring with a Teflon paddle or stainless steel spoon

for 10 minutes.

6. ORGANICS - Using a stainless steel spoon, carefully place 250 cc of sediment in

a 500 ml glass bottle for chemical analysis. Do not overfill container; adequate

head space must be maintained to allow for sample expansion if frozen at later date.

CARE MUST BETAKEN TO ASSURE THAT THE INSIDE OF THE BOTTLE,

BOTTLE CAP, AND THE SAMPLE ARE NOT CONTAMINATED. Record the

sample number, wrap the jar in “bubble wrap” to protect it from breakage, and

place the sample on ice (NOT dry ice). To reduce the possibility of breakage, the

sample should be stored at 4EC, NOT FROZEN.

7. METALS - Using a stainless steel spoon, place approximately 100cc of sediment

into a pre-cleaned plastic (HDPE) sampling jar. Record the sample number and

keep on ice at 4EC.
26

8. Total Organic Carbon - Using a stainless steel spoon, place approximately 100cc

of sediment into a pre-cleaned glass sampling jar. Record the sample number and

keep on ice at 4EC.

9. SEDIMENT GRAIN SIZE - Using a stainless steel spoon, place approximately

100cc of sediment into a clean plastic (HDPE) sampling jar. Record the sample

number and keep on ice at 4EC. Store this sample on ice (NOT dry ice). The sample

must be stored at 4EC, NOT FROZEN.

10. SEDIMENT TOXICITY - Using the stainless steel spoon, fill approximately 75-

85% of the 1 gallon plastic container for toxicity testing with sediment (minimum

volume required is 3000 ml). Record the sample number on the bottle, and place

the sample on ice (NOT dry ice). The sample must be stored at 4EC, NOT

FROZEN.

6.4 Quality Control/Quality Assurance

6.4.1 Chemistry samples

There are a number of steps that can be taken to ensure the integrity of the samples collected.

1. The interior surfaces of the grab sampler (including the underside of the hinged top)

must be washed with a laboratory-grade detergent and thoroughly rinsed prior to

use to assure that no sediment remains from the previous station.

2. Prior to use, all Teflon and stainless steel supplies which are to come into contact

with samples must also be properly cleaned. Once washed, crews must take

precautions to assure that they do not become contaminated (e.g., by laying the

stainless steel spoon on the deck).

3. As soon as any of the stainless spoons or bowls begin to rust they should be

discarded. Equipment made from high-quality stainless steel will reduce the rate at

which equipment needs to be replaced.

4. ASSURE THAT THE PROPER LABELS (e.g., BAR CODES) ARE AFFIXED TO

ALL SAMPLES.

5. Excess seawater should be carefully drained from the surface of the grab by

“cracking” the sampler slightly or siphoning off the water.

6. All grabs used in the composite must meet the criteria for an acceptable grab. It is

especially important to make sure that the surface sediments did not wash out of the

sampler.
27

7. Care should be taken to assure that the sediment saved for chemical and

toxicological analyses is collected only from the top two cm of the grab.

8. Care must be taken to assure that the chemistry samples do not become

contaminated. This requires great care in extracting the sample, homogenizing it,

and placing it in the proper container. Because of the potential for contamination,

the chemistry samples should be the first ones removed from the homogenate. If it

is raining when the sample is collected, all activities should be conducted under a

tarp to prevent contamination of the sample by rain water.

9. Great care must be taken to avoid atmospheric contamination from engine exhaust.

The boat engine must be turned off or the boat maneuvered to assure the engine

exhaust is down wind of the sample.

10. Exposure of the sample to the atmosphere should be minimized. Whenever

possible the sample should be covered because contamination from the atmosphere,

even without the engines running, can be significant.

11. Samples should be placed in a cooler on ice as soon as they are collected and

recorded.

12. The grab must be suspended off the deck at all times to avoid contamination.

13. If the vessel is unable to anchor, the position relative to station should be monitored

carefully during benthic colletion.

6.4.2 Benthic biology

Field crews must assure that all grabs processed are acceptable according to the criteria

described above, and that no organisms are lost during any step, including transferring the sample

to the sieve, and during sieving. Also, samples must be properly identified and preserved to

assure they are received by the processing laboratory in acceptable condition.

6.4.3 GRAIN SIZE

Samples collected for grain size analysis require no special QA steps other than carefully

following the directions discussed earlier and assuring proper storage. Note that grain size

samples must NOT be frozen.

6.4.5 TOXICITY

Since sediment toxicity samples are collected from the same homogenate used for

sediment chemistry, the steps outlined above should be followed. In addition, because of the

possibility of failure of a toxicity test, it is important that a full 3 L of sediment be collected for

analysis at each station. This will provide a sufficient volume of sediment for re-testing if
28

necessary. NOTE: Toxicity sample must not be frozen.

6.5 Safety Considerations

All sediment grab samplers are dangerous pieces of equipment. Once the device is

cocked, it could accidentally trip at any time. The operators must be careful not to place hands or

fingers in a position where they could be damaged (or amputated) in the event that the device

trips prematurely.

The sampler is a heavy piece of equipment (especially when full). At all times, the

operators must take care when deploying or retrieving this gear; under adverse weather

conditions, extreme caution is mandated.
29

SECTION 7

FISH TRAWLS

After all required sediments are collected, one or more trawls are made to collect fish for

species composition, relative abundance, chemical analysis, and pathological examination.

Many states already have their own trawling protocols. Described below are the protocols for

Coastal 2000. Existing state protocols and gear may be substituted following discussions with

the C2000 Field Coordinator.

7.1 Gear and General Protocols

A fish trawl is a funnel-shaped net that filters fish from the near bottom waters. Fish are

herded by ground wire and doors into the mouth of the funnel where fish are captured. The basic

components of a trawl net are described briefly below. The actual specifications of the net used

by AED for EMAP are found in Appendix B. The following discussion describes the use of a

net with 13.5-m footrope, that used by the NE field teams; however, a smaller trawl with a 5-m

(16-ft) footrope is employed for the shallow conditions routinely encountered in other regions

(e.g., the gulf and southeast estuaries).

The doors of the net provide spreading power to the net. Water pressure against the doors

force them to spread the wings of the trawl. The wings are the beginning of the webbing and

form the mouth of the funnel on two sides of the net. The wings are bordered on top and bottom

by a headrope and a footrope, respectively. For a single warp rig, each end of the headrope, or

top line, is attached directly to the upper ring on the back of the doors. Each end of the footrope,

or bottom line, is attached to the bottom ring of the doors. For strength and weight, a sweep is

attached to the footrope. At the bosom, or top of the curve of the mouth, the wings attach to the

body of the net. The top portion of the body has an overhanging panel, or square, which prevents

fish from escaping over the top panel of the trawl. Continuing back toward the terminus of the

net are the first and second bellies which are normally symmetrical top and bottom. The bellies

contribute most of the body of the net, and therefore make up most of the taper. The cod-end is

the rear portion of the trawl net which serves as a collecting bag for all that is captured by the

trawl.

Fish are collected using a high rise sampling trawl with a 13.5-meter footrope with a

chain sweep. Tow duration is 10 minutes with a towing speed of 2-3 knots against the prevailing

current. Speed over the bottom should be 1-3 knot. Fish are sorted and enumerated, examined

for evidence of gross pathological conditions, and selected specimens retained and properly

processed for tissue chemical analysis. Subsampling of fish is conducted as necessary. The

outline below describes the specific protocol to be followed during trawling operations. The

procedures include: net deployment, vessel operation while under tow, net retrieval, and

processing.
30

Types of trawls can be defined as follows:

STANDARD TRAWL - This trawl is the “quantitative” trawl performed at all stations for

community structure and abundance determination. Only one standard trawl should be

performed at EVERY station. Any fish sample type can be taken from a standard trawl. Fish

are identified, measured, and examined for pathological conditions.

NON-STANDARD TRAWL - At selected stations non-standard trawls may need to be

performed following the completion of a standard trawl only to obtain a sufficient number of

fish for tissue chemistry.

The type of fish samples that will be collected are as follows:

Pathology Fish - These are fish observed by the field crew to have a gross external pathology

(lump, growth, ulcer, fin rot, gill erosion, and/or gill discoloration). ALL species are examined

for external pathology, therefore, pathology fish may be of any species collected. Pathology

fish are collected only during the standard trawl. Any fish found with one of these conditions

is preserved in Dietrich’s fixative for confirmation by a specialist. These fish are Pathology

fish.

Taxonomy QA fish - Fish that cannot be identified in the field are to be sent back to the

appropriate agency for identification by an expert taxonomist.

7.2 Trawl Preparation

(Portions of these instructions are specific to 24-26' EPA boats. Modifications may be

necessary depending upon the vessel used for trawling)

1. Inspect the trawl net for holes, including cod-end liners, and mend/replace as necessary

prior to departure from the dock. Inspect all hardware for wear and replace as needed.

All connections should be made securely and tightened with a wrench. Do NOT rely on

hand tightening shackles, bolts, or other fasteners.

2. Lead the winch wire from the drum through the turning block on the mast assembly and

through the snatch block at the end of the boom.

3. Attach the bridle to the winch wire with a shackle. Wind both legs of the bridle onto the

main winch drum, while maintaining tension on the wire. All bridle connections should

be tightened with a wrench.

4. Arrange the net on the deck with the cod-end aft and the head rope on top. Close the end

of the cod-end by using a cod-end knot. Check to make sure there is no escapement

possibility through the cod end rings. The line should pass through the rings at the back

of the cod end and around the net just in front of these rings. Coil the float line from the
31

cod-end to the float, and position it on the net for easy access.

5. Attach the legs of the net to the trawl doors. The top leg of the net is the extension of the

headrope and must be secured to the top aft ring of the door. The bottom leg is the

extension of the sweep and must be secured to the bottom aft ring of the door. One bridle

wire should be attached to each door at the towing point of the chain harness. Shackles

should be used for all connections.

7.3 Net Deployment

(Portions of these instructions are specific to 24-26' EPA boats. Modifications may be

necessary depending upon the vessel used for trawling)

1. After all preparation steps have been completed, the chief scientist or captain should

check all resources available (chart, navigational aids, land marks etc.) to determine that

there are no under water hazards. Determine the direction of current flow and survey the

probable trawl track for potential hazards, such as other vessels, deployed commercial

fishing gear (nets, pots, etc.), shallow water, or unsuitable substrate. In addition, depth,

weather, and sea conditions should also be evaluated prior to each trawl. The decision as

to whether or not to collect a sample is the responsibility of the chief scientist or captain.

2. With the starboard engine in neutral, the boom should be positioned out over the

starboard gunnel with a enough incline for the doors to clear the rail. Lead the bridles

through the snatch block on the boom, raise the doors with the winch, and bring them to

rest on the gunnel (starboard door forward, port door aft). Circle the boat slowly to

starboard. When the starboard side is down current, deploy the float and safety line

attached to the cod-end. Flake the net into the water from the cod-end to the wings.

Check to make sure that the legs of the net are not twisted before continuing deployment.

Pay out wire until the doors are well behind the engines. Swing the boom to the

centerline then lower the boom, releasing tension on the snatch block (the wire should

now be on the goalpost assembly). Head slowly into the current (e.g., 1 knot) and

continue to pay out wire until appropriate warp length is obtained (consult Table 4 for the

proper amount of wire to be released based on water depth). Great care should be taken

to prevent fouling of the propeller with the net. Care should also be taken to maintain

tension on the tow warp to avoid fouling the net on bottom. The starboard engine can be

engaged when the gear is clear of the props and the doors spread.

7.4 Trawling

1. As soon as the required warp length is reached, the winch operator should inform the

captain that the net is ready for towing. The captain then visually resurveys the trawl

track, records the time, initiates the trawl clock, records the start coordinates, and begins

the tow. An attempt should be made to trawl along a uniform depth contour.
32

2. Boat speed should be 2-3 knots. Speed over bottom, as measured by GPS position,

should be between 1 and 3 knots. If it becomes apparent that these conditions will not be

met, the net should be retrieved and a different trawl direction tried.

Table 4. Amount of Winch Wire to be Used for Trawling

Water depth (ft) Ratio of line to water depth Line out (including the 125' bridle)

10 (3 m) 7:1 Bridle only (38m)

20 (6 m) 7:1 Bridle+20' (44m)

30 (9 m) 7:1 Bridle+60' (56m)

40 (12 m) 6:1 Bridle+120' (75m)

50 (15 m) 5.5:1 Bridle+155' (85m)

60 (18 m) 5:1 Bridle+180' (92m)

70 (21 m) 4.6:1 Bridle+202' (100m)

80 (24 m) 4.2:1 Bridle+216' (104m)

90 (27 m) 3.8:1 Bridle+222 (106m)
33

3. During the trawl tow, the captain should monitor the depth finder for potential

obstructions or sudden changes in depth. If a hazard is identified or a hang up occurs, the

net should be retrieved and another tow attempted approximately 100 m from the initial

trawl track. If three unsuccessful attempts are made, or 1.5 hours effort is expended,

trawling operations should be aborted. If a successful 10 minute trawl cannot be

accomplished, fish can still be collected from a shorter trawl for chemistry.

4. The duration of all standard trawls should be 10 minutes from the time the pay-out of

warp is completed until the time hauling begins.

7.5 Net Retrieval

(Portions of these instructions are specific to 24-26' EPA boats. Modifications may be

necessary depending upon the vessel used for trawling)

1. After approximately 10 minutes of trawling, record the end coordinates then haul back

the wire until approximately 10 meters of the bridle is still out.

2. Put the starboard engine in neutral. Throttle back and raise the boom so the wire clears

the goal post assembly. Turn the boat slightly to starboard and move the boom over the

starboard side (the boom should be controlled by the vangs during this process).

3. Take in wire until the doors are at the block. Haul the cod end in by hand or use the

capstan head to assist. Retrieve the float line and float.

7.6 Safety Considerations

Operation of the trawl can be a dangerous operation. In addition to the dangers of using

the winch and capstan, improper towing procedures could capsize the boat. The net should

always be towed off the stern, with the winch cable passing through the towing bracket. Towing

off the side of the boat can capsize it. Care must also be taken when pulling the net in over the

side. If the net is full, the total weight may be too great to use the mast and boom.

When deploying the net, the crew must be careful not to entangle themselves or other

gear in the net, bridle, or winch cable. This could result in serious personal injury or damage to

equipment.

All trawling operations must be conducted in a manner consistent with maintaining the

safety of the crew. The captain will determine when weather or sea conditions are unsafe for

trawling.

In the event of net hang-ups on bottom obstructions, the captain must consider the safety

of the crew before attempting to free the gear. A means to sever the tow line should be
34

immediately available to the crew during all trawl operations. SEVERING THE LINE SHOULD

ONLY BE PERFORMED AS A LAST RESORT AND WHEN THE LINE IS SLACK!!!

SEVERING IT WHILE UNDER TENSION COULD RESULT IN WHIPLASH OF THE LINE

AND SEVERE PERSONAL INJURY.

Before deploying the trawl, the captain should ensure that other vessels do not present a

safety hazard during the tow. Whenever possible, the captain shall contact nearby vessels by

marine radio to make them aware of the trawling operation. In addition, the marine radio should

be monitored by the crew prior to and during trawl operations. Appropriate day shapes must be

flown.

7.7 Criteria for Voiding Tows

A standard tow will be considered void if one or more of the following conditions occur:

1. A tow cannot be completed because of hang down, boat malfunction, vessel traffic, or

major disruption of gear.

2. Boat speed or speed over bottom is outside the prescribed, acceptable range.

3. The cod-end is not tied shut.

4. If the tow continues for more than two minutes beyond the ten-minute tow duration, or

is discontinued less than eight minutes following the start.

5. The net is filled with mud or debris.

6. A portion of the catch is lost prior to processing.

7. The tow wire, bridle, headrope, footrope, or up and down lines parted.

8. The net is torn (>30 bars in the tapered portion, >20 bars in the extension or cod end, or

multiple tears that, in the opinion of the chief scientist, may have significantly altered

the efficiency of the net).

7.8 Endangered Species

All species considered to be rare, threatened, or endangered should be processed

immediately and released alive. At the discretion of the chief scientist, photographs may be

taken to document the catch.

7.9 Sample Processing

Once a catch is brought on deck, fish are identified to species, measured, counted,

examined for external pathology, and processed for chemical analysis.
35

7.9.1 General Processing

1. After all fish have been sorted, process fish for pathological examination as described

below. Sampling for pathology and Chemistry are performed concurrently with the

collection of composition and abundance data. Only fish, lobster, and blue crab are

recorded. Other invertebrates and trash are noted in the datasheet then discarded.

2. Measure, with a measuring board, the fork length to the nearest millimeter, of individuals

of each species. If there are fewer than 30 individuals of a species, all individuals should

be measured. If it is estimated that more than 30 individuals of a species were caught, a

subsampling procedure should be used to measure between 30-50 individuals.

Subsampling will be accomplished by randomly selecting fish from the buckets. All data

are entered onto data sheets and later into the computer.

NOTE -

Dog fish - stretched total length

Skates - total length

Rays - wing tip to wing tip, and total length

Unforked - total length without extraneous filaments

Blue crab - carapace width

Lobster - Carapace length

3. Enter data on the fish data sheets. Common names are preferred.

4. All fish not measured for length (i.e., those subsampled) are counted, either by direct

count or weight-counts. When extremely large catches of schooling fish such as bay

anchovy or other clupeids are made, abundance may be estimated by weight-counts. At

least 100 individuals should be weighed in a batch, and 2 batches should be weighed to

determine mean weight per individual. All remaining fish should be weighed, and the

total number of fish estimated and recorded on the data sheet. If two or more obvious

size classes are present in a sample (e.g., young-of-year and adults), the size classes

should be treated as separate species for the purpose of counting.

5. After all processing has been completed, the chief scientist should review the trawl data

sheet for discrepancies and inaccuracies. When any questions have been resolved, he/she

signs the data sheets as being reviewed and the remaining portion of the catch can be

returned to the water. When significant mortality occurs and the trawl site is in a highly

visible area, the captain may elect to retain the catch until more discrete disposal can be

accomplished. Under no circumstances should the crew give fish away to the general

public.
36

7.9.2 Processing of Fish for Gross External Pathological Analysis

I. Gross examination of fishes:

All individuals collected from a standard trawl will be identified and counted, and the

first 30 individuals of each species will be measured. All individuals measured (i.e. the first 30)

that exceed 75 mm in length will be examined for evidence of gross external pathology (lumps,

growths, ulcers, fin rot, gill erosion, and gill discoloration). The examination is intended to be a

rapid scan of the surface of individuals to be completed while other fish measures are being

completed (i.e., identifying, enumerating, measuring). This scan should take no longer than 10-

15 seconds per fish. Fish determined to show evidence of a pathology are assigned a sample

number and processed appropriately (see below). The type of pathology will be noted on the data

sheet. These are PATHOLOGY FISH. Only fish collected in “standard” trawls are saved

for pathology.

II. Selection, killing and fixation for transfer:

Proper fixation of specimens is critical to the ultimate quality of the data obtained. Fish

should be examined and fixed while still alive or shortly after death (within one hour of

collection). Specimens should not be frozen or kept on ice at any time.

A. All specimens with gross lesions or other suspect conditions, as identified in Section I

above, will be processed and coded individually. All these fish will be transferred as

indicated below (Section III) to EPA’s Gulf Ecology Division (GED) for subsequent

examination.

1. Carefully cut the entire length of the abdominal cavity open using scissors or a sharp

knife. Gently insert the instrument into the abdomen near the anus and make an

incision to the operculum. Cut with a lifting motion so that the incision is made

from the inside outward, taking care not to injure the visceral organs. Remove the

lateral musculature from one side of the animal’s visceral cavity to facilitate the

fixation of the internal organs. Remove the opercula, and immerse in fixative (see

step 4).

2. If the total length of the fish exceeds 15 cm, only a portion of the fish will be saved

for laboratory analysis. Carefully cut, through the entire thickness of the fish, from

the top of the operculum back along the spine, until a position behind the visceral

cavity is reached, and then a 90E change in direction towards the anus. The head

and viscera are then saved. Remove both opercula, and musculature covering the

visceral cavity on one side. The head and thorax can be separated at the esophagus

if needed. Any abnormalities found on the remaining portion of the fish (which is to

be discarded) are excised along with the surrounding tissue, and saved with the head

and visceral cavity. For fishes smaller than 15 cm, the entire fish is saved. (See

Figure 3).
37

3. If an external growth is present, slice through the lesion with one clean cut using a

sharp razor blade.

4. Place the sample (whole fish or head, visceral cavity and abnormalities excised) in

an “onion bag” or a plastic zip lock bag with multiple perforations. Assign an

appropriate sample number to each fish, affix the bar code to a fish tag, and attach

the tag to the fish. Record this number on the data sheet, along with all other

pertinent information on that fish. Place the bag in a tight sealing plastic container

with sufficient fixative to completely cover the specimen. Specimens should be

fixed in Dietrich’s fixative for one or two days.

Dietrich’s Fixative (to make ~5 gals.)

37-40% Formaldehyde or 100% formalin --------------- 1500 ml

Glacial Acetic Acid ------------------------------------------- 300 ml

95% Ethanol ------------------------------------------------- 4500 ml

Distilled water ----------------------------------------------- 9000 ml

5. Carefully record pertinent information relating to each individual sample on

the data sheet.

III. Shipping of preserved specimens:

Fish should soak in Dietrichs Fixative for at least two days prior to shipment. To ship,

wrap the fish in cheesecloth dampened with Dietrichs. Place the wrapped fish in several

layers of airtight plastic bags and pack in cardboard boxes or coolers. No specific

temperature criteria apply.

7.9.3 Tissue Chemistry
38
39

1. For the two most abundant taxa designated as target species, measure and retain nine

individuals within the desired size range for chemical analysis. Individuals are

randomly selected from all those collected until nine of the appropriate size have

been selected. If no individuals in the primary size range were collected, those that

are closest to the preferred size range are selected.

NOTE:

a. Even if a tow is voided for species composition and abundance, fish collected

can still be processed for chemistry.

b. Target species for tissue analysis are designated on a regional or state-bystate basis. If no target species are caught from a site but other species are taken,

a surrogate target can be selected from the best available by-catch; attributes that

define a good target include: demersal, high tropic level, representative of area,

and adequate size or age class.

2. Record on the datasheet the size, species, sample number (see Step 3), and any other

appropriate notes.

3. Place one bar code on the data sheet. Place the twin bar code on a plastic tag and

affix to the fish by placing the twist-tie through the mouth and out the operculum.

4. Wrap individual fish in aluminum foil (with the tag exposed), place all fish of that

species in a single zip-lock bag, affix the “composite” bar code, and place it in a

cooler on DRY ICE.

5. All samples should be placed immediately on DRY ICE for freezing. When adding

new samples to the cooler containing the dry ice, samples should be rearranged to

assure that these samples are in contact with the dry ice so they will freeze rapidly.

One option would be to use one cooler for freezing fish, and a second for storing

them. This is dependent on the equipment carried on the boat, and therefore, the

amount of space available. If freezing on-board is not practical, fish must be stored

on ice until the crew reaches the dock. The time before freezing should be

minimized.

6. Repeat trawling (standardized methods not required) for up to 11⁄2 hours if needed to

obtain at least five individuals of at least one target species. Fish collected in these

trawls are processed for chemistry only.
40

7.10 Quality Assurance

In order for the net to “fish” properly, the proper amount of winch cable must be let out.

Consult Table 4 for the proper scope. Care must also be taken to assure that fish are not lost

from the net during retrieval.

It is important that the tow time and speed be as close to the desired values as possible.

Any deviations should be noted on the data sheet.

It is important not to contaminate fish which are saved for chemical analysis. Every

effort should be made to keep them from coming in contact with very dirty surfaces. It is

especially important to ensure that no cuts are made into the flesh.

Table 5. Listing of Northeast Region Target Species for Chemical Analysis (sizes are the target

sizes for fish saved for chemical analyses).

SPECIES SIZE RANGE (mm)

Catfish Species

 Channel Catfish 200-300

 White Catfish 200-300

Scup 70-115

Summer Flounder 350-450

Weakfish 300-400

Winter Flounder 100-200

Blue Crab 120-170

Lobster

7.11 Contingency Plans

Considering the wide variety of environments to be sampled by C2000, it is likely that

towing a net will be impossible at some stations. If, due to repeated snags, a successful trawl

cannot be performed within 1.5 hours of starting, no further attempts should be made. This is

noted on the data sheet.

In the event that a “standard” trawl cannot be obtained because of space limitations, the

crew can still use either alternative gear to collect fish and shellfish for chemistry. This may

include purchasing lobster or crab from local fishermen. The preferred method would be to

accompany the fishermen during the collection to ensure the crabs or lobster are collected in the

proximity of the station. It is important that the crew is sure the lobsters or crabs were
41

collected in the vicinity of the station.

7.12 Collection Permits

Many states require scientific collection permits for the collection of fish using trawls.

Permits issued for C2000 activities must be carried on each boat. A permit must be presented to

any appropriate state official that requests to see it.
42

SECTION 8

PACKAGING AND SHIPPING SAMPLES

After samples are collected, following proper packaging and shipping procedures are

critical steps in assuring the integrity of the samples. Failure to follow these procedures could

result in the loss of valuable data. Each sample type requires different handling as described

below. Packaging and shipping are to be performed within several days of sample collection.

Samples may be sent either to an approved state lab or one of the Coastal 2000 “national

labs.” This determination is made well before the start of the sampling season. The protocols

described below are specific to samples being sent to the national labs; however, it is

recommended that samples being sent to local labs be processed similarly.

 As samples are packaged for final shipment, the sample number of each sample must be

recorded. This can be done on a hard-copy shipment form, or electronically entered into a

computer. Upon completion of packaging, a unique tracking number (bar code label) is affixed

to the SIDE of the cooler, and this number is also recorded on the shipment form. The number

must be placed on the side, not top, so as not to interfere with the carrier’s tracking system

(which also may use bar codes). In addition to the carrier’s air bill, a mailing label should also be

affixed to the cooler as an additional precaution against loss. A packing list must accompany the

shipment. This can be a photocopy of the completed shipment form, or a printout from the

computer. Samples that are “hand-carried” require the same paperwork (less the air bill) and

tracking as those shipped by commercial carrier. Since coolers need to be shipped back to the

crew, a return air bill should also be included in the cooler.

All samples, except those preserved in formalin or Dietrichs, are shipped overnight.

Shipping should only take place on Mondays through Wednesdays, otherwise samples will

arrive at the analytical laboratory on the weekend when there may be no one available to

accept them.

The C2000 Field Coordinator must be informed each time a shipment is sent out. The

information needed includes the name of the analytical laboratory, the shipment ID number, the

carrier’s air bill number (this is especially important), and a list of the samples included in the

shipment.

8.1 Proper Packaging Methods

Proper packaging of samples is critical in assuring they arrive at the receiving laboratory

in good condition. Improper packaging can result in damaged or lost samples. This is costly in

terms of time and money. There are several important aspects of proper packaging: assembly of

the shipping box (if required), the amount of blue or dry ice needed, and proper packaging of the

contents.
43

Each team should be supplied with several sizes of coolers. The appropriate size should be

selected to minimize “dead” space.

Each team carries coolers with dry ice and blue ice to keep samples frozen or cool prior to

shipment. Blue ice blocks can be frozen by placing them under the dry ice, or in a freezer for

crews operating out of a home base. For dry ice, a general rule of thumb is at least 5-10 pounds,

with another pound for every pound of sample (ASSUMING THE SAMPLES ARE ALREADY

FROZEN). The amount of blue ice needed to keep samples cool is approximately one pound per

pound of sample. This should guarantee samples arrive frozen or cool (depending on the ice

type) even if the shipment is delayed a day. Frozen samples must always be shipped on dry ice,

and refrigerated samples must always be shipped on blue ice packs.

Because of the need to ship fish and crabs frozen, relatively large amounts of dry ice will

be needed.

In addition, it is recommended that the sample be sandwiched between refrigerant, i.e. dry

ice should be packed both above and below the sample. It is also important that the cooler

contain a minimum of air space. Therefore, packing material should be inserted above the top

ice layer to fill the cooler.

A third consideration for all sample types (not just cooled or frozen samples) is proper

packaging within the shipping cooler. While packing a shipment cooler, one should assume that

the cooler will be improperly handled. All samples should be protected and sufficient packing

material included to eliminate any possible movement of the samples within the cooler. All

material that could possibly leak, such as water or sediment samples, should be sealed with

sealing tape and packaged in zip lock bags. All glass sample bottles should be bubble wrapped

and sealed in a zip lock bag. Any whirl paks should also be taped to ensure the metal tabs do not

puncture adjacent bags, and placed in a zip lock bag.

Proper storage and shipment conditions are summarized in Table 6. Federal Express no

longer requires a Dangerous Materials waybill for all shipments of formalin in concentrations of

10% or less. Federal Express does, however, require a class 9 placard, UN number, packing

description and an emergency phone number for all shipments of dry ice.
44

Table 6. Sample holding and shipping conditions

SAMPLE TYPE HOLDING CONDITIONS SHIPPING CONDITIONS

Sediment Biota Preserved in Formalin Once per Week

Sediment Grain Size Refrigerated * Ship Weekly

Sediment Organics Refrigerated * Ship Weekly

Sediment Metals Refrigerated * Ship Weekly

Sediment TOC Refrigerated * Ship Weekly

Sediment Toxicity Refrigerated * Ship Weekly

Chlorophyll Filter Frozen on Dry Ice * Ship Weekly

Total Suspended Solids Refrigerated * Ship Weekly

Dissolved Nutrients Frozen on Dry Ice * Ship Weekly

Fish Chemistry Frozen on Dry Ice * Ship Weekly

Pathology QA Wrapped in Cheesecloth Once per Week

* Crews should attempt to ship as frequently as logistically possible. Samples must be shipped at

least once per week.

8.2 Benthic Biology Samples

Samples for benthic community analyses are preserved in formalin in the field. These

samples are in plastic containers with tight fitting screw-top lids. As these samples are

preserved, there is no need to keep them cool. Shipment boxes should not weigh more than 50

pounds. The lid of each jar should be checked to assure that it is tight, and the lid taped with

sealing tape. The bar code label of each container is then read and the samples placed in an

insulated shipping box. The insulation is for protection rather than thermal regulation. As

described above, a computer printout of the sample numbers included in this shipment is

enclosed in the box.

The box is then sealed and an appropriate shipping label affixed. Be sure to pack all

bottles upright, and to fill gaps with packing material. Overnight delivery is not required.

This shipment contains formalin; however, since the final concentration is 10% or less,

Federal Express no longer requires a Dangerous Goods Airbill and Shipper Certification form.

8.3 Sediment Chemistry Samples

Following collection, sediment samples for TOC, organics, and metals chemical

characterization should be refrigerated rather than frozen because freezing greatly increases the

likelihood of breakage of the glass container. It is also recommended that samples be shipped
45

cool, but not frozen, for the same reason.

Sediment chemistry samples should be shipped at least weekly, preferably early in the

week. Sample bottles should be wrapped in bubble wrap to protect them from breakage, and

sealed in a plastic zip lock bag. They should then be placed in an insulated cooler with an

appropriate amount of blue ice.

Sediment chemistry samples must be shipped Next Day Service to the appropriate lab.

8.4 Sediment Toxicity Samples

Sediment samples collected for sediment toxicity testing must be kept refrigerated (4EC),

NOT FROZEN. Sample must be shipped at least weekly. Of all the samples to be analyzed, the

sediment toxicity samples are the most “time-critical” due to the nature of the testing and the

relatively short permitted holding time (<28 days). Containers are then placed upright, along

with an appropriate amount of blue ice, in an insulated box or cooler. Sediment toxicity samples

are shipped Next Day Service to the appropriate lab.

8.5 Grain Size Samples

Samples for grain size analysis are collected along with each sample collected for benthic

biology and sediment chemistry/toxicity analyses. Samples for grain size analysis should be kept

cool (4EC), but not frozen. If these samples are contained in Whirl Packs sealed with metal

wraps, tape should be placed around the ends of these wraps at the time of collection to prevent

the metal tips from piercing one of the other bags. Samples should be shipped weekly. Whirl

Packs should be placed in a ziplock bag and packed into an insulated box or cooler with an

appropriate amount of frozen blue ice to keep the samples cool. Place a thin layer of paper

between the blue ice and the Whirl Paks to keep them from freezing.

Samples should be shipped Next Day Service to the appropriate lab.

8.6 Chlorophyll, Nutrient, and Total Suspended Solids Samples

Samples should be shipped at least weekly. Chlorophyll and nutrient samples should be

frozen and shipped on dry ice. TSS samples are shipped on blue ice.

Samples should be shipped Next Day Service to the appropriate lab.

8.7 Fish Chemistry

Samples should be shipped at least weekly. Samples must be frozen and shipped on dry

ice.

Samples should be shipped Next Day Service to the appropriate lab.
46

8.8 Pathology QA Samples

These samples are preserved in Dietrich’s fixative. Fish must be well preserved in

Dietrichs Fixative prior to shipment. The fish should be removed from the bucket of fixative,

wrapped in multiple layers of Dietrichs-soaked cheesecloth, then placed in multiple layers of

airtight plastic bags. Samples then should be packaged into cardboard boxes or coolers and

shipped to the appropriate lab. Overnight service is not required.

8.9 Instructions for FEDEX Shipping with Dry Ice

A. Use Regular Airbill

1. Sender’s Section: Fill in the Date and Your Name.

2. Confirm the recipient’s name, shipping address and phone number

3. Payment Section: Confirm that Bill Sender Box (#1) is checked

4. Services Section: Check Priority Overnight Box /your packaging, leave Freight

Service and Instructions sections blank

Delivery and Special Handling Section: Check Dry Ice Box and Fill in the total

weight of dry ice for the shipment.

5. Complete section 6 of the Airbill.

Note: print the weight of the dry ice in Kg, not the total box weight!

B. Proper Labeling of Shipping Boxes containing Dry Ice

1. Stick Number 9 placard label (available from FEDEX) so that clearly visible on side

of box

2. On the same side of the box the following information must be printed:

Dry ice, 9, UN 1845, ___box X ___ Kg , 904 III

Dangerous goods - Shipper’s declaration not required

Note: print the weight of the dry ice in Kg, not the box! Up to 2.2 kg permitted

(5 lbs).

3. Be sure that each box has an Address Label with the correct address of the receiving

facility.
47

APPENDIX A

List of Suggested

Supplies and Equipment
48

This list provides a complete (more or less) listing of recommended supplies and equipment for Coastal

2000 sampling in the northeast states. It is intended as an aid in planning efforts.

General

Boat set up appropriately for sampling

Appropriate safety gear

Navigation equipment - GPS or Differential GPS (preferred), depth finder

Vehicle to move people around as needed

Vehicle to store gear in, including shipping coolers (only needed for crews traveling away from a

base location)

Communications equipment between shore and boat (e.g., cellular telephones, VHF...)

Computer for data entry (laboratory or portable)

Bar code reader (recommended)

Bar codes for all samples - EPA can coordinate

Datasheets - EPA can provide templates

Pre-labeled station datasheet packages

Data entry software (e.g., JetForm’s Formflow Filler for EPA datasheets)

Shipping containers (e.g., coolers)

Shipping labels

FEDEX (or other carrier) airbills

Shipping Bar codes

“Blue ice”

Coolers for storing samples (both frozen and chilled)

Field notebooks

Water-resistant paper for datasheets (e.g., Rite-in-the-rain)

Waterproof pens for writing on datasheets

Clipboards

Insulated gloves for handling dry ice

Bubble wrap for shipping

Shipping scale for weighing packages

“Packing list enclosed” envelopes

“This side up” labels

Class 9 placards for dry ice shipments

Strapping tape for shipping

Duct tape for everything else

Buckets and/or hose and washdown pump

Meter stick

Waterproof markers

Paper towels

Kimwipes
49

Latex or other gloves for handling contaminated sediments or formalin

Scissors

Suggest pingers for overboard gear in case it is lost

Water Quality Monitoring

Profiling instrument to measure depth, temperature, salinity, pH, and DO (e.g., Hydrolab

DataSonde4, YSI 6000, etc., with appropriate deck unit and cabling as needed)

Back-up/QA instrumentation:

Additional DO meter (unless doing Winklers)

Thermometer

Refractometer

pH standards

Salinity standards

Spare DO membranes & electrolyte

Spare parts for profiler

Batteries

Light (PAR) or transmissometer with appropriate deck unit or datasheets and cables

Secchi disk with marked line

Water sampling bottle for nutrients

Filtration apparatus for Chlorophyll

a) 2- 47mm filter holders

vacuum manifold

4 liter overflow bottle

12vdc vacuum pump or hand pump or

b) stainless steel, 25 mm filter holder

standard luerlock syringe

47 or 25 mm GF/F filter pads (2 per sample, up to 6 per station)

Clean 60cc nalgene bottle for nutrients (3 per station)

1-L Nalgene for TSS samples (3 per station)

Storage containers for filters and other supplies

MgCO3

Filter forceps

Graduated cylinders, 250, 100, 50, 10 ml

DI water for rinsing

Squirt bottles

Aluminum foil for wrapping samples (suggest pre-cut squares from Thomas Scientific)

Whirlpaks or ziplock bags for foil-wrapped filter pads

Dry ice for freezing samples
50

Sediment sampling

0.04m2

 Young-modified Van Veen grab sampler (or other)

Grab stand

Weights for grab (several)

Pads for grab (several)

0.5 mm stainless steel sieve

Sieve box

“Tub” or bucket for dumping sediment into

High-quality stainless mixing pot, with lid, for sediments (2)

Stainless spoons (several)

500 cc glass jars for organics (Ichem pre-cleaned)

250 cc HDPE jar for metals (Ichem pre-cleaned)

125 cc glass jar for TOC (Ichem pre-cleaned)

125 cc HDPE jar for grain size

4-L HDPE jar for toxicity (pre-cleaned)

1-L Nalgene for benthic infauna (3 per station plus spares)

Electrical tape for sealing lids of benthic containers

Formaldehyde (formalin)

Rose Bengal stain

Borax (can get at supermarket)

Centimeter ruler

Wide mouth funnel

Squirt bottle

Alconox

Scrubbing brushes

Fine forceps for picking worms from screen

Fish Sampling

13.5 m otter trawl (several)

Doors for otter trawl

Bridles

Timer

Fish measuring board

Heavy duty aluminum foil

Waterproof tags with tie strings

Taxonomy keys

Heavy duty dissecting scissors

Scalpel or sharp razor blade

Cheesecloth
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5-gallon bucket for Dietrich’s fixative

LARGE ziplock bags for fish composites

Onion bags

Dietrich’s fixative

Formalin

Glacial Acetic acid

95% ethanol
52

APPENDIX B

Trawl Net

Specifications
53

C2000-NE (EMAP) Trawl Net Specifications

 3:1 trawl net of 3" webbing

 Headrope length = 13.5 meters

 Sweep = 16.5 meters

 Hanging line and headrope of 1⁄2” poly dacron with thimbles spliced into ends

 Up and down lines of 1⁄2” poly dacron spliced into the headrope and hanging line

 Webbing of 3" #21 twisted polyethylene (= European #312 twisted stranded) reinforced

along the mouth frame with gussets

 Headrope flotation of 4 small (5", 760 grams buoyancy) plastic floats

 Codend of 11⁄2” #24 nylon, 64 wide by 65 deep

 Sweep of 3/16" chain with 12 feet of 1⁄4” chain along the mouth

 Bridle is 125', 1⁄4” stainless steel wire
54
55

APPENDIX C

Coastal 2000 Field Data

Forms
56

Gulf Region Field Data Sheets

The following field data sheets have been designed by GED for the Gulf region Coastal

2000 effort. Their use is recommended but not required. GED can provide electronic templates

for these forms. Included are the following data sheets (sample forms are attached):

Station Form

Hydrographic Profile

Water Sample Collection

Benthic Infauna Collection

Sediment Sample Collection

Fish trawl

Fish Data
57
58
59
60
61
62
63
64

APPENDIX D

Water Quality

Measurements

by

Hydrolab® DataSonde 3/

Surveyor 4

(Routinely Utilized in Northeast Region)
65

 WATER QUALITY MEASUREMENTS BY HYDROLAB® DATASONDE 3/

SURVEYOR 4

This section describes the procedures for the calibration and operation of the Hydrolab

DataSonde 3 datalogger (referred to as either “Hydrolab” or “DataSonde”) attached to a Surveyor

4 deck unit. This is the equipment used by AED field crews to perform water column profiles for

the parameters of dissolved oxygen, salinity, temperature, pH, and depth. The protocols

following are excerpted from past EMAP-Virginian Province field manuals. Similar procedures

should be followed by field crews using other instruments.

Included in this section is the operation of a YSI model 58 DO meter. Protocols require a

duplicate measurement, using a different instrument or method, at each station for the purpose of

Quality Assurance. Winkler titrations are recommended for crews experienced with this

procedure. If this wet chemistry method is not practical, a duplicate surface measurement can be

made with a separate properly-calibrated DO meter. For the data to be acceptable, both values

must agree to within 0.5 mg/L.

D.1 Hydrolab Datasonde3® unit

Obtaining a vertical profile of the water column using a Hydrolab Datasonde3® or similar

unit is one of the first activities performed at every station. A Hydrolab is a sophisticated

instrument designed to collect high-quality data for salinity, temperature, dissolved oxygen (DO)

concentration, pH, and water depth. At each station the instrument will be used as a CTD

(instrument that measures Conductivity, Temperature, and Depth - in this case, also measures pH

and DO) to obtain a vertical profile of water column conditions. Training of all personnel

expected to operate this instrument is necessary to assure reliable operation and acceptable data.

Below are general instructions for calibrating and deploying these units.

D.1.1 Setup and Calibration

The following is a brief summary of the calibration of the Hydrolab. The manual should be

referred to for detailed instructions and should be read prior to calibration. During calibration,

the Datasonde unit should be attached to the gel pack battery to conserve the unit’s internal

battery supply. Calibration should be performed every morning prior to the start of sampling.

1. To calibrate the Hydrolab Datasonde units, the software package “Procomm” will be

used. Attach the DataSonde unit to the computer with the data cable, making sure the

computer is reading from the correct port (com 2 on AED “Rocky” laptops) and at a

baud rate of 9600bps. The first time you calibrate the Hydrolab, you will have to select

the parameters that C2000 will be using and remove any other parameters (in order to

save memory and battery power). Once you are in Procomm, lines of data will be

displayed (if this is not the case check the battery or refer to the manual). Pull up the

menu by depressing the space bar. To select the parameters hit ‘P’.
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2. You can now add the following parameters by hitting the letter in parentheses for each

parameter and then choosing E for (E)nable. The parameters that need to be added are:

(p)H

(S)alinity

D(O)

(%) Sat

(D)epth/Level

(B)attery

3. Then remove the following parameters by choosing (D)isable:

Specific (C)onductance/Resistivity

(R)edox

D.1.2 Calibration of the salinity sensor

The salinity sensor will be calibrated against a sample of seawater that has a known salinity

[from a high quality laboratory salinometer calibrated with IAPSO Standard Seawater (a.k.a.

“Copenhagen” water)]. The Hydrolab will always be equipped with the salt water cell block.

Rinse the sensor & calibration cup 3 times with a small amount of the salinity standard (shaking

vigorously with the calibration cap in place). Fill the calibration cup to within a centimeter of the

cup’s edge and make sure there are no bubbles in the conductivity cell block. From the Calibrate

menu, choose (S)alinity and enter the standard value in parts per thousand.

D.1.3 Calibration of the pH sensor

Rinse the sensors and calibration cup thoroughly with deionized water prior to and following

filling the cup with the standard pH buffers. Fill the calibration cup with the pH 7 standard

buffer. Wait until the reading stabilizes to hit the space key, access the calibrate menu and enter

the pH value. Now finish calibrating the pH sensor using the pH 10 standard. It is important that

there is not a lot of drift in the measurement before entering the calibration standard value. If you

cannot get the readings to stabilize, it is time to clean the sensors and check the battery power.

D.1.4 Calibration of the Dissolved Oxygen sensor

The calibration of the dissolved oxygen sensor is highly sensitive to the maintenance of the

sensor itself. To make the calibration process go more smoothly, it is important to examine the

DO membrane and make sure it is has not dried out, become damaged or dirty, that there are no

bubbles in the electrolyte, and that you have waited at least 12 hours (preferably 24 hours)

after changing a membrane to calibrate.

With the unit turned upside down, fill the calibration cup with ambient room temperature tap

water (or DI) to the O-ring line on the DO sensor, making sure all of the sensors have been well

rinsed prior to this. Tightly put the calibration cap on and shake the unit to aerate the water.
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Remove the cap on the calibration cup and, using the corner of a kimwipe, remove all water

droplets from the membrane surface. Put the calibration cap on (upside down). Wait for the

readings to stabilize, and then depress the space key to access the calibration menu. Enter

760mm for the barometric pressure (if a barometer is available the exact pressure can be entered,

but the range at sea level has only a minimal effect on the calculated DO reading), and then enter

the DO percent saturation (100% for the standard membrane).

D.2 Obtaining Hydrolab Profile

 At each station, the general procedures for collection of data are as follows:

1 Connect the Hydrolab to the end of the winch cable with a shackle and TIGHTEN THE

PIN. Make sure a “pinger” is attached to the unit. A 50 pound weight should be hanging

approximately 0.5 meter below the unit, and one float (sufficient buoyancy to float the

Hydrolab without the weight) attached to the top. This will prevent the unit from impacting

the bottom.

2. Remove the protective cover from the probes and connect the stirrer.

3. Connect the unit to the Surveyor 4 deck unit and initialize logging.

4. Connect the stirrer to the upper bulkhead connector.

5. Lower the unit over the side and allow it to equilibrate at the surface for at least two minutes

after the unit begins logging.

6. While the unit is equilibrating, lower a YSI probe (see Section 4.3) with stirrer over the side

to the same depth as the Hydrolab. Record the reading from the YSI on the CTD datasheet.

This serves as a Quality Control check on the operation of the Hydrolab. A surface salinity

and temperature should also be obtained with a refractometer and the YSI meter or

thermometer, respectively.

Make sure that the Hydrolab surface readings agree with those from the QC check (e.g., the

DO readings must agree to within 0.5 mg/L). If they agree, continue with the cast. If they do

not agree, recalibrate the YSI and obtain another surface reading. If they then agree, continue

with the cast. If they do not, try another Hydrolab.

7. Lower the Hydrolab according to the following schedule:

Shallow sites (# 2 m) - every 0.5 m interval;

Nominal depths (>2<10 m) - 0.5 m (near-surface) and every 1-m interval to nearbottom (0.5 m off-bottom);

Deep sites (>10 m) - 0.5 m (near-surface) and every 1-m interval to 10 m, then at 5-m

intervals, thereafter, to near-bottom (0.5 m off-bottom).
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Allow the unit to stabilize at each stop during descent. Save the data from each depth on the

Surveyor unit and record the values on the Hydrographic Data Sheet. Once the weight hits the

bottom the unit will float 0.5 meter above.

8. Repeat the process on the upcast.

9. Connect the Surveyor to the computer and download the data (this can be done back at

the dock). The file should be saved as “XX00xxxxctd.csv” where XX00xxxx is the

station number (e.g., MA000001ctd.csv).

D.3 YSI Model 58 Dissolved Oxygen Meter and Probe

The YSI will be used to take oxygen measurements at the surface as a Quality Control

check on the Hydrolab. The following information details the maintenance and operation of

the YSI Model 58 Dissolved Oxygen Meter.

D.3.1 Initial Setup of The YSI

1. The YSI Model 58 has two separate sets of batteries, one for the oxygen meter and the

other for the stirrer. Both sets consist of 4 D-size Alkaline batteries. These are accessed

by removing the four screws on the back panel then carefully pulling the meter back

away. The upper battery holder is for the oxygen meter, the lower holder for the stirrer

batteries. Note that the stirrer batteries will probably require more frequent replacement,

whereas the meter batteries will most likely last throughout the entire field season.

Observe correct polarity whenever changing batteries in either holder.

2. When the YSI meter batteries are low, the LOWBAT warning will show continuously on

the display (the LOWBAT warning may flash momentarily as the meter knob is turned

off, but this is normal). The initial appearance of LOWBAT indicates about 50 hours of

meter battery life. The normal life for the meter batteries is about 1000 hours.

To check the YSI stirrer batteries, turn and hold the STIRRER knob to the BATT CHK

position. If the LOWBAT warning shows continuously on the display then the stirrer

batteries should be changed. The initial appearance of the LOWBAT warning in the

BATT CHK position indicates 5 hours or less of stirrer battery life. The normal life for

the stirrer batteries is about 100 hours.

3. While the meter is still open, observe the position of the sliding switch in the upper right

hand corner of the meter. This switch sets the meter sensitivity for the type of membrane

on the oxygen probe. The switch should be in the middle position, set for a 1 mil

(“standard”) membrane.

4. Close the meter housing and gently tighten the corner screws. DO NOT OVER

TIGHTEN these screws, as they are easily stripped. As you close the meter, work the
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rubber gasket so that the outer edge overlaps both halves of the housing.

D.3.2 Changing the YSI Probe Membrane

The procedure for changing the YSI probe membrane is similar to that for the Hydrolab

membrane. However there are some differences, so it’s important to be familiar with

both procedures. The YSI membrane should be changed weekly, or sooner if the probe is

difficult to calibrate or is slow to respond. Visual inspection is the best indication of

when to change the membrane: if the membrane is fouled, wrinkled, cut, has bubbles

underneath it, or the gold cathode is tarnished...then it’s time. Try to schedule membrane

replacement at the end of a field day, or the night before. This allows the membrane more

time to “relax” and equilibrate.

1. Prepare the electrolyte by dissolving the KCl crystals in the dropper bottle with distilled

water. Fill the bottle to the top.

2. Unscrew the sensor guard, and remove the O-ring and membrane. Rinse the sensor with

distilled water and then with electrolyte. Gently wipe the gold cathode ring with a

kimwipe or paper towel.

3. Fill the sensor with electrolyte. If you’re right-handed, grasp the sensor in your left hand

with the pressure compensating vent to the right. Successively fill the sensor body with

electrolyte, then pump the diaphragm with the ERASER end of a pencil or with some

similar soft, blunt tool. Continue filling and pumping until no more air bubbles appear.

Tap the sensor with the pencil to free any bubbles trapped on the sides.

4. Remove a membrane from the “standard membrane” package (DO NOT use the Hydrolab

membranes - they are different). Secure the membrane under your left thumb. Add a few

more drops of electrolyte to the sensor to form a meniscus over the gold cathode.

5. With the thumb and forefinger of your other hand, grasp the free end of the membrane.

6. Using a continuous motion, stretch the membrane UP, OVER, and DOWN the other side

of the sensor. Stretching forms the membrane to the contour of the probe.

7. Secure the end of the membrane under the forefinger of the hand holding the probe.

8. Set the O-ring on the membrane above the probe, and using your thumb and index finger,

roll the O-ring down over the probe until it is seated. Try not to touch the membrane

surface while doing this. Gently tug at the exposed corners to remove all wrinkles, then

trim away the excess membrane below the O-ring and replace the sensor guard. Inspect

the membrane to make sure there are no bubbles, wrinkles, or cuts.

9. The probe should be stored in the open-ended plastic bottle provided for that purpose.

Moisten the sponge or paper towel in the end of the storage bottle to prevent the
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membrane from drying out. The membrane needs to relax for a minimum of 12 hours following

installation.

D.3.3 Calibration of the YSI Oxygen Meter

The YSI should be calibrated before sampling at EACH station, and the meter and

attached probe should be turned on for at least 10 minutes prior to calibration or

sampling. In practice this means turning the meter on at the beginning of the day and

leaving it on (with the possible exception of very long transit periods between stations).

On field days when the probe is not being used leave the meter in the % switch positions

(or, in the case of the model 57, in the 0-10 MG/L position).

1. Calibration will be done in the probe storage/calibration chamber. Confirm that a moist

piece of towel or sponge is present in the bottle. Remove any water droplets from the

membrane surface by drying with the corner of a paper towel.

2. Set the function switch to ZERO, and when the display reading has stabilized, readjust

display to read 0.00.

3. Reset the function switch to % mode. When the display reading has stabilized, unlock

the O2 CALIB control locking ring and adjust the display to read 100%. Relock the

locking ring to prevent inadvertent changes. Avoid exposing the calibrated probe to large

thermal changes, such as from direct sunlight or lying on a hot deck.

D.3.4 Operation of the YSI Oxygen Meter

In general the YSI will be used to confirm the proper operation of the CTD.

1. Calibrate the YSI (See above; Section 4.3.3).

2. Remove the storage/calibration chamber and the sensor guard, and CAREFULLY screw

the probe into the stirrer. The probe membrane should NOT touch the stirrer blades.

Membrane damage occurs most often when the probe is being inserted or removed from

the stirrer. If a measurement isn’t to be taken immediately, wrap the stirrer-probe unit in a

moist towel and set it out of the sun.

3. Set the function switch to 0.01 MG/L mode.

4. To perform a surface YSI check place the probe next to the CTD DO probe with the

stirrer ON. Set the YSI salinity from refractometer reading. Record temperature from a

thermometer, and DO from the YSI on the “CTD CAST DATA SHEET”.

5. If using the Hydrolab to obtain a bottom water dissolved oxygen concentration:

A. Collect a bottom water sample in the GO-FLO bottle.
71

B. Draw out a small sample from the bottle and measure the salinity using the

refractometer. Set the SALINITY switch to this value, and record the salinity on

the “CTD CAST DATA SHEET”.

C. Prop open the Go-Flo bottle. A Hydrolab sensor guard without the weight works

well for this.

D. Insert the stirrer-probe unit into the GO-FLO bottle and turn the stirrer ON.

E. When the meter reading has stabilized, record the oxygen value on the “CTD

CAST DATA SHEET”.

F. Remove the probe, turn the stirrer OFF, rinse the probe with freshwater, replace the

storage bottle, and store the unit out of sunlight.