United States
Environmental Protection
Agency
Office of Research and
Development
Washington DC 20460
EPA/620/R-01/003
June 2001
National Coastal
Assessment
Field Operations
Manual
Environmental Monitoring and
Assessment Program
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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, Rl 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
Recycled/Recyclable
Printed with vegetable-based ink on
paper that contains a minimum of
50% post-consumer fiber content
processed chlorine free.
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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. Environ-
mental 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 i. 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
IV
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SECTION :
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:
• sediment contaminant concentrations
• sediment toxicity (Ampelisca abdita)
• benthic species composition
sediment characteristics (grain size, organic carbon content, percent water)
• water column dissolved nutrients
• chlorophyll a concentrations,
• total suspended solids concentration,
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surface and bottom dissolved oxygen, salinity, temperature, and pH
water clarity
contaminant levels in fish and shellfish
external pathological condition of fish
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 C2000is 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., 00° 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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Arrive on Station
Record Coordinates
Record Weather & Sea Conditions
& Presence of Trash, SAV, Macroakjae
Determine Seccni Depth
Collect Water Column Profile
And Light Measurements
Collect Appropriate Water Samples
For Chlorophyll, Nutrients, & TSS at
Surface, Mid, and Bottom
Collect Sediment for Community Analysis
& Chemistry, Toxicity, and Grain Size
(Biology Grabs Interspersed Among Chem/Tox)
Conduct Fish Trawl(s)
Filter for Chlorophyll
Save Filter
n
Save Filtrate for
Dissolved Nutrients
Collect 1 liter sample for TSS
1 Whole Grab for
Benthic Community
Surficial Sediment Homogenized
Identify, count, measure,
& examine fish
TOC I Organics I I Inorganics I Toxicity II Grain Size I
Figure 1. Flow chart of sampling activities conducted at C2000-NE stations.
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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:
Where,
SSYY-XXXX
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:
MAOO-0001 to MAOO-0035
TXOO-0001 to TXOO-0035
CAOO-0001 to CAOO-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., RIOO-0001 for Rhode Island, year 2000, station 1) followed by
a sample type code. The list below uses station RIOO-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
CTD Cast
Light measurement (PAR) profile
Surface chlorophyll
Surface suspended solids
Surface dissolved nutrients
Mid-depth chlorophyll
Mid-depth suspended solids
Mid-depth dissolved nutrients
Bottom chlorophyll
Bottom suspended solids
Bottom dissolved nutrients
Benthic infauna(l)
Benthic infauna(2)
Benthic infauna(S)
Sediment Toxicity
Sediment grain size
Sediment Organics
Sediment Metals
Sediment TOC
Standard fish trawl
Non-standard fish trawl
Fish chem. species 1 composite
Fish chem. species 1 individuals
Fish chem. species 2 composite
Fish chem. species 2 individuals
"Other" sample type 1
"Other" sample type 2
OuaJitv Assurance Samples ffull
Fish pathology QA
Chlorophyll QA
Dissolved nutrients QA
TSSQA
Type Code
CTD
PAR
SCL
SSS
SN
MCL
MSS
MN
BCL
BSS
BN
BI1
BI2
BI3
ST
SG
SO
SM
oc
STRL
TRL
FC1
FC1-1 toFCl-9
FC2
FC2-1 to FC2-9
OTH-1
OTH-2
Example Sample Number
RIOO-0001 -CTD
RIOO-0001 -PAR
RIOO-0001 -SCL
RIOO-0001 -SSS
RIOO-0001 -SN
RIOO-0001 -MCL
RIOO-0001 -MSS
RIOO-0001 -MN
RIOO-0001 -BCL
RIOO-0001-BSS
RIOO-0001 -BN
RIOO-0001 -BI1
RIOO-0001 -BI2
RIOO-0001 -BI3
RIOO-0001 -ST
RIOO-0001 -SG
RIOO-0001 -SO
RIOO-0001 -SM
RIOO-0001 -OC
RIOO-0001 -STRL
RIOO-0001 -TRL
RIOO-0001 -FC1
RIOO-0001-FCl-l
ToRIOO-OOOl-FCl-9
RIOO-0001 -FC2
RIOO-0001 -FC2-1
ToRIOO-0001-FC2-9
RIOO-0001 -OTH-1
RIOO-0001 -OTH-2
Bar-Coded?
N
N
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
N
N
Y
Y
Y
Y
Y
Y
range of sample numbers eiven for C2000-Northeastf
PATH
CL
N
SS
QAOO-0001-PATH
To QAOO-0500-PATH
QAOO-0001-CL
To QAOO-0300-CL
QAOO-0001-N
To QAOO-0300-N
QAOO-0001-SS
To QAOO-0300-SS
Y
Y
Y
Y
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11
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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 DataSondeS/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, alien 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)
p.H ± 0.3 units
salinity ±1.0 ppt
temperature ± 1.0 °C
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 mis 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
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the temperature value appearing on the deck display as the "MEASURED" value for TEMP; the
measured temperature should be within ±1 °C 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 sees); 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 O-
ring); 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
14
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saturated with moisture (this system of calibration is commonly referred to as water saturated-air
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-
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inch PVC piping. Cap one end of a 24-in section of pipe with a plug and affix the resultant well
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 CaE/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 f>2<10 m) - 0.5 m (near-surface) and every 1-m interval to near-bottom (0.5
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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).
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.
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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 rn) - 0.5 m (near-surface) and 0.5 m off-bottom1
Standard sites f>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 4°C 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,
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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.
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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 4°C 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.
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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
21
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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 floe, 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.
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Acceptable grab
At least 7 cm deep with even surface
Unacceptable grab
Sloping surface
Unacceptable grab
Insufficient volume
Unacceptable grab
Wash-out
Unacceptable grab
Overfilled
Figure 2 Illustration of acceptable and unacceptable grabs for benthic community analysis. An
acceptable grab is at least 7 cm in depth (using a 0.04m2 Van Veen sampler), but not
oozing out of the top of the grab, and has a relatively level surface.
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7.
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.
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.
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.
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 fstained and buffered)
80
80
Rose Bengal stain
Borax
100% formalin
1/4 teaspoon
8 heaping tablespoons
two gallons
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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 4°C, 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 4°C, NOT FROZEN.
7. METALS - Using a stainless steel spoon, place approximately lOOcc of sediment
into a pre-cleaned plastic (HDPE) sampling jar. Record the sample number and
keep on ice at 4°C.
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8. Total Organic Carbon - Using a stainless steel spoon, place approximately lOOcc
of sediment into a pre-cleaned glass sampling jar. Record the sample number and
keep on ice at 4°C.
9. SEDIMENT GRAIN SIZE - Using a stainless steel spoon, place approximately
lOOcc of sediment into a clean plastic (HDPE) sampling jar. Record the sample
number and keep on ice at 4°C. Store this sample on ice (NOT dry ice). The sample
must be stored at 4°C, 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 4°C, 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.
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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
27
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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.
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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.
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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
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cod-end to the float, and position it on the net for easy access.
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.
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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)
20 (6m)
30 (9m)
40 (12m)
50 (15m)
60 (18m)
70 (21 m)
80 (24m)
90 (27m)
7:1
7:1
7:1
6:1
5.5:1
5:1
4.6:1
4.2:1
3.8:1
Bridle only (38m)
Bridle+20' (44m)
Bridle+60' (56m)
Bridle+120' (75m)
Bridle+155' (85m)
Bridle+180' (92m)
Bridle+202' (100m)
Bridle+216' (104m)
Bridle+222(106m)
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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
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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.
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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.
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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 90° 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).
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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.
DI. 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
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Fish less than 15 cm in length
Cut end of operculum
to expose gills
Remove abdominal
covering on one side
Fish greater than 15 cm in length
Excess to be discarded
unless a lesion is present
Figure 3. Description of how to expose interior organs for proper preservation of Pathology Fish.
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1. • For the two most abundant taxa designated as target species, measure and retain nine
individuals Xvithin 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-by-
state 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 Wi 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.
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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
White Catfish
200-300
200-300
Scup
Summer Flounder
Weakfish
Winter Flounder
Blue Crab
Lobster
70-115
350-450
300-400
100-200
120-170
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
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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.
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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.
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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 iri 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.
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Table 6. Sample holding and shipping conditions
SAMPLE TYPE
HOLDING CONDITIONS SHIPPING CONDITIONS
Sediment Biota
Sediment Grain Size
Sediment Organics
Sediment Metals
Sediment TOC
Sediment Toxicity
Chlorophyll Filter
Total Suspended Solids
Dissolved Nutrients
Fish Chemistry
Pathology QA
Preserved in Formalin
Refrigerated
Refrigerated
Refrigerated
Refrigerated
Refrigerated
Frozen on Dry Ice
Refrigerated
Frozen on Dry Ice
Frozen on Dry Ice
Wrapped in Cheesecloth
Once per Week
* Ship Weekly
* Ship Weekly
* Ship Weekly
* Ship Weekly
* Ship Weekly
* Ship Weekly
* Ship Weekly
* Ship Weekly
* Ship Weekly
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
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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 (4°C),
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 (4°C), 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.
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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 Ibs).
3. Be sure that each box has an Address Label with the correct address of the receiving
facility.
46
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APPENDIX A
List of Suggested
Supplies and Equipment
47
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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, VHP...)
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
48
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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
MgC03
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
49
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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-cleane'd)
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
50
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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
51
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APPENDIX B
Trawl Net
Specifications
52
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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 Vi" poly dacron with thimbles spliced into ends
Up and down lines of Vi" 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 1V&" #24 nylon, 64 wide by 65 deep
Sweep of 3/16" chain with 12 feet of 1A" chain along the mouth
Bridle is 125', W stainless steel wire
53
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44
25
70
65
AP
22
25
70
65
Figure B1. Diagram of EMAP trawl net. Headrope is 13.5 meters, sweep is 16.5 meters.
54
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APPENDIX C
Coastal 2000 Field Data
Forms
55
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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
56
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STATION FORM
Coastal 2000 - Gulf Region
STAGING AREA
STATION DEPTH (ft.): _
STATION'NAME I
»AV
GPS
CREW
WEATHER
HABITAT
TYPE
DATE:ftIMff>iyW)
TIMKZ<3N&OEul
fn*-'£iCeiKisl
S.AT: ««>.(»)
ARRIVAL T|ME:(I!H:MM)
Dfl-AKTtSMKIHHMM)
tON-.WJW.no,
3f VCX txptaci in'NOTES'
CAPIAIS.
CitgW 1 :
0»K*'2:
VISITOK !^
VISITOR 5:
VISITOR 1:
DTid*! River
D Open Water
D B»v ou/lolct
n Inli-r-Tidal
D KockwShcll liflllom
Dco»IR«f
ClMjmii
dOyiltrBtd
D Marina ,
f^| (Zeuss Bed
D Other
SAV I Ptesem ? Q Yes o
MARINE
DEBRIS
Piesertt?
O Vtis or O ^*°
Type
D GLASS
D WOOD
O PLASTIC
O CANS
O Other, . .
1' .
MEOSSED BY:
. COMPUTER BNTRV 8V:_
57
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HYDROGRAPHIC PROFILE
Coastal 2000 - Gulf Region
QC CHECK
TIME:
-------
WATER SAMPLE COLLECTION
Coastal 2000 - Gulf Region
DATE:
(MMIJOYW
„ FILTRATION MRTHOB: Q SYRINGE orQ VACUUM
SURFACE (OJim)
NUTRIENT
S 1 (Filleted Sample - 60 m!)
1 1 OS mm Filter} Volume Filtered: (ml)
[_J {Uttfiltcied Sampie • ! Lter)
MID-DEPTH
NUTRIENT
fm
CHLOROPHYLL
(MCI,)
TSS
(1HSS)
N/A (<2 m) LJ
U (RlEBd Sample - 60 ml)
LJ OS mm Filter) Volume Filtered: (ml)
1 ...1 {If nfilhred Sample - 1 Lite)
BOTTOM (0.5 off
bottom)
NinrRlE!fT
(Bfi)
CttlOBOfUYU.
(SCL)
TSS
(BSS)
1 1 (Filtered Sample - 60 ml)
1 i (25 mm Filter) Volume Fitlcrtd: (mli
D (Unnitered Sarapfe - 1 LUer)
* Sxmple }ahel to be pbccd
-------
BF.NTHIC INFAUNA COLLECTION
Couilal 2000 - Gulf Regtan
UAIT.:IMMUDYY)_
_ CRAB T VPE: O v» v«» or O fo
SAMPLE NO. i
tnin-
TI)HE:
DESCRIKnON:
NO.OFJAKStfSEU:
DEPTH (cm):
SAMPLE.NO. Z
TIME (I!I1:.MM):
INSCRIPTION:
NO. OF JARS USED:
UKPTH (cm):
SAMPLE MO. 3
(Blif
TIMK (IU1:MM):
DESCRIPTION:
NQ.OFJARSVSEU:
|pKPTU(a»):
mple lubtl In bt plwd »« nntuhKr i< U»l«i i« U«»it4l
COMCUIER tVI'Kf BVl,.
60
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SEDIMENT SAMPLE COLLECTION
Coastal 2000 - Gulf Region
DATE:(MMDDYY)_
Station Name
SEDIMENT CHEMISTRY I—| Organics (SO)' (Glass Jar)
U.D
Metals (SMJ {Nalgene 125 nil)
TOXICITY (STJ
(1 ga! -Nalgene)
GRAIN SIZE
-------
FJS1I TRAWL #
Coastal 2000 - Gulf Region
Net Width (ft):
Station Name
TRAWL INFO
DATE (mm/tld/yy):
HELMSMAN:
LINE OUT (m):
TRAWL START
LAT (WOO.OO1):
LON (00°00.00'):
HEADING IN DEGREES MAGNETIC:
START TIME (HH-.MM):
TRAWL END
LAT <00°00.00'):
LON <000OOJ)I)'):
END TIME (HH:MM):
TRAWL DETAIL
TRAWL TAKEN: o Yes or o No
IF NO, EXPLAIN:
TRAWL SUCCESSFUL: o Yes or o No
IF NO, EXPLAIN:
ANYTHING CAUGHT: o Yes or O No
RECORDER:
COMPUTER ENTRY BY:
62
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FFSH DATA
Coastal 2000 - Gulf Region
DATE:
Station Name
COMMON NAME: •
OENUS SPECIES NAME
Fisli
Number
I
2
3
4
5
6
7
8
- 9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
Fish
Length
Samples
Chemistry Pafholosv •
Pathology
Type
TRAWL INFO:
NUMBER
TYPE
0 STANDARD
o NON-STANDARD
0 OTHER. . .
PATHOLOGY
OBSERVATIONS
G - GILL ABN
U - ULCERS
L - LUMPS/BUMPS
S - SKELETAL ABN
E - EYE ABN
TOTAL COUNT:
RECORDER:
ENTRY BY;
63
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APPENDIX D
Water Quality
Measurements
by
Hydrolab® DataSonde 3/
Surveyor 4
(Routinely Utilized in Northeast Region)
64
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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.I Hydrolab DatasondeS® unit
Obtaining a vertical profile of the water column using a Hydrolab DatasondeS® 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.I.I 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'.
65
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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(0)
(%) Sat
(D)epth/Level
(B)attery
3. Then remove the following parameters by choosing (D)isable:
Specific (Qonductance/Resistivity
(R)edox
D.I.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.I.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.I.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.
66
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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 near-
bottom (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).
67
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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 "XXOOxxxxctd.csv" where XXOOxxxx is the
station number (e.g., MAOOOOOlctd.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 LOWB AT warning will show continuously on
the display (the LOWB AT warning may flash momentarily as the meter knob is turned
off, but this is normal). The initial appearance of LOWB AT 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 B ATT CHK
position. If the LOWB AT warning shows continuously on the display then the stirrer
batteries should be changed. The initial appearance of the LOWB AT warning in the
B ATT 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
68
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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 KC1 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.
6.
7.
8.
9.
With the thumb and forefinger of your other hand, grasp the free end of the membrane.
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.
Secure the end of the membrane under the forefinger of the hand holding the probe.
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.
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
69
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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.
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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.
71
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United States
Environmental Protection Agency/ORD
National Health and Environmental
Effects Research Laboratory
Research Triangle Park, NC 27711
Please make all necessary changes on the below label,
detach or copy, and return to the address in the upper
left-hand corner.
If you do not wish to receive these reports CHECK HEREEH;
detach, or copy this cover, and return to the address in the
upper left-hand corner.
PRESORTED STANDARD
POSTAGE & FEES PAID
EPA
PERMIT No. G-35
Official Business
Penalty for Private Use
$300
EPA/620/R-01/003
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