&EPA
United States
Environmental Protection
Agency
Office of Water
(WH - 556F)
EPA 842-B-92-004
September 1992
Monitoring Guidance for the
National Estuary Program
Final
printed on recycled paper
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MONITORING GUIDANCE FOR THE
NATIONAL ESTUARY PROGRAM
Final
Office of Water
Office of Wetlands, Oceans, and Watersheds
Oceans and Coastal Protection Division
U.S. Environmental Protection Agency
Washington, D.C. 20460
September 1992
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Executive Summary
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Purpose This document provides the National Estuary Program (NEP) with guidance on how to
design, implement and evaluate a monitoring program. The careful use of this docu-
ment will lead to more complete information to measure the success of management
actions initiated under the Comprehensive Conservation and Management Plan
(CCMP) as well as essential data to evaluate environmental risks and trends. It could
also be useful to other coastal and marine managers with monitoring responsibilities.
The National In 1987 Congress amended the Clean Water Act and created the NEP to recognize
Estuary Program nationally significant estuaries, to protect and improve their water quality, and to
enhance their living resources. The Environmental Protection Agency (EPA) has
selected 18 estuaries under the NEP and has plans to add more.
One of the major products of each estuary program is the CCMP which identifies
priority problems, determines environmental quality goals and objectives, and speci-
fies action plans and compliance schedules to ensure that designated uses of the
estuary are protected. Included in the CCMP is a NEP monitoring plan.
National A recent report by the National Research Council (Managing Troubled Waters, The
Importance Role of Marine Environmental Monitoring; NRC, 1990) recognized the need for vast
improvements in marine monitoring. Separately, the Expert Panel on the Role of
Science at EPA established by the EPA Administrator in 1991 challenged the EPA to
acquire and to use the best scientific information in the decision making process of
EPA program offices (Safeguarding the Future: Credible Science, Credible Decisions;
The Report of the Expert Panel on the Role of Science at EPA). This guidance docu-
ment incorporates the recommendations of both national reports. Furthermore, the
EPA Science Advisory Board commended this guidance for its systems approach to
monitoring and the use of objective-driven criteria in designing monitoring programs.
Highlights A monitoring program should meet three interrelated objectives: verify the predicted
responses of the estuary to management actions, measure the effectiveness of pro-
grams implemented under the CCMP, and provide essential information that can be
used to redirect and refocus the CCMP.
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To meet these objectives, five steps are outlined in the monitoring design framework:
Step 1 Develop Monitoring Objectives and Performance Criteria
Explicit statements of the monitoring objectives and perfor-
mance criteria with which to measure monitoring program
success are crucial. Example monitoring objectives, perfor-
mance criteria, and tools for developing the objectives are
presented.
Step 2 Establish Testable Hypotheses and Select Statistical Methods
Establishing testable hypotheses ensures that the results of the moni-
toring program will be unambiguous and that the objectives of the
program can be met. To guarantee sufficient information and the right
type of information, the NEPs should choose the statistical model that
will be used to analyze the field data and testable hypotheses prior to
the collection of any samples. References on the use of various
statistical methods are provided.
Step 3 Select Analytical Methods and Alternative Sampling Designs
Detailed specifications for each monitoring variable of the monitoring
program must be developed. These include field sampling methods,
laboratory procedures, and QA/QC procedures. Alternative sampling
methods are described, and the use of data from existing monitoring
programs is discussed.
Step 4 Evaluate Expected Monitoring Study Performance
During the course of the monitoring program design and execution,
performance evaluations provide a systematic procedure for measur-
ing success in terms of the ability to meet program goals. Statistical
tools for evaluating program performance are described, and the use of
feedback mechanisms are recommended to refine objectives and to
improve monitoring program performance.
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Step 5 Implement Monitoring Study and Data Analysis
Data management and data analysis, key issues that are often over-
looked in the design of monitoring programs, are as important to the
success of the monitoring effort as the collection and laboratory
analysis of field data. Data and information management issues are
addressed, and the importance of the analysis and the communication
of program results are discussed.
Two case studies from existing estuarine monitoring programs (i.e., Puget Sound
Ambient Monitoring Program and Chesapeake Bay Monitoring Program) are pre-
sented which demonstrate the process of developing a strategy and the use of statistical
methods to evaluate the monitoring plan before and after implementation. Also,
sampling and analytical methods available for monitoring estuarine water quality,
sediment quality, biological resources and human health risks are presented.
NEP's environmental monitoring activities depend on integrating state, federal and
local activities. The NEP is encouraged to coordinate with existing monitoring efforts
such as the EPA's Ecosystem Monitoring and Assessment Program (EMAP), the
National Oceanic and Atmospheric Administration's National Status and Trends
Program, and the U.S. Geological Survey's National Water Quality Assessment
Program.
A companion to this document (Technical Characterization Guidance for the National
Estuary Program) will be released soon. The characterization guidance will help
NEPs identify and set priorities among problems within the estuary during CCMP
development.
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Acknowledgments
For their help in the development and review of this document, special thanks
are extended to the following individuals: Tom Armitage and Joe Hall who
served as project managers at different times; John Armstrong of EPA Region
10; members of EPA's Science Advisory Board, members of the National
Oceanic and Atmospheric Administration's Interagency Ecosystems Monitoring
Workgroup, and program directors and staff of National Estuary Program
Management Conferences.
Technical support for development of this document was provided by Tetra
Tech, Inc. under EPA Contract No. 68-C1-0008.
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VII
Table of Contents
1.0 Introduction 1
1.1 Background 3
1.2 Recommended Monitoring Design Procedures 7
1.3 Monitoring Program Management 11
2.0 Develop Monitoring Objectives and Performance Criteria 17
2.1 Monitoring Program Objectives 18
2.2 Performance Criteria 22
2.3 Additional Guidance 23
3.0 Establish Testable Hypotheses and Select Statistical Methods 25
3.1 Establish Testable Hypotheses 25
3.2 Selection of Statistical Methods 28
4.0 Select Analytical Methods and Alternative Sampling Designs 31
4.1 Selection of Field and Laboratory Methods 31
4.2 Alternative Sampling Designs 32
4.3 Use of Existing Monitoring Programs 35
5.0 Evaluate Expected Monitoring Program Performance 43
5.1 Evaluate the Expected Performance of Individual Monitoring Program Components 44
5.2 Evaluate Overall Program Performance 45
5.3 Statistical Power Analysis Methods , 45
6.0 Design and Implement Data Management Plan 51
6.1 Data Management 52
6.2 Data Analysis 53
7.0 Communicate Program Results 55
8.0 References 59
Appendices
A Case Studies
Al.O The Puget Sound Ambient Monitoring Program Case Study A-3
Al.l Purpose and Approach A-3
A1.2 Development of PS AMP: Institutional Arrangements A-4
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A1.3 The Puget Sound Ambient Monitoring Program A-9
A1.4 Implementation of PSAMP and Cost A-13
A1.5 Summary and Recommendations A-19
A1.6 References A-23
A2.0 Chesapeake Bay Monitoring Program: Detection of Trends in Estuaries A-25
A2.1 Purpose and Approach A-25
A2.2 Chesapeake Bay Program A-25
A2.3 Evaluation of Monitoring Program Performance A-29
A2.4 Use of Power Analysis Results , A-40
A2.5 References ;A-40
B Methods
B.I Methods - Introduction B-3
B.I Methods Chapter Format B-3
B.2 Quality Assurance/Quality Control(QA/QC) Considerations B-8
B.3 Statistical Design Considerations B-ll
B.4 Literature Cited and References B-14
Bl.O Water Column Physical Characteristics B-17
Bl.l Rationale B-17
B1.2 Monitoring Design Considerations B-17
B1.3 Existing Analytical Methods B-19
B1.4 QA/QC Considerations B-24
B1.5 Statistical Design Considerations B-27
B1.6 Use of Data B-28
B1.7 Summary and Recommendations B-28
B1.8 Literature Cited and References B-31
B2.0 Water Column Chemistry .B-35
B2.1 Rationale B-35
B2.2 Monitoring Design Considerations B-35
B2.3 Existing Analytical Methods ; B-38
B2.4 QA/QC Considerations B-42
B2.5 Statistical Design Considerations B-45
B2.6 Use of Data B-46
B2.7 Summary and Recommendations . B-46
B2.8 Literature Cited and References B-49
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IX
B3.0 Sediment Grain Size B-53
B3.1 Rationale B-53
B3.2 Monitoring Design Considerations B-53
B3.3 Existing Analytical Methods B-58
B3.4 QA/QC Considerations B-59
B3.5 Statistical Design Considerations B-61
B3.6 Use of Data B-61
B3.7 Summary and Recommendations B-61
B3.8 Literature Cited and References B-63
B4.0 Sediment Chemistry B-65
B4.1 Rationale B-65
B4.2 Monitoring Design Considerations B-65
B4.3 Existing Analytical Methods B-71
B4.4 QA/QC Considerations B-74
B4.5 Statistical Design Considerations B-79
B4.6 Use of Data B-79
B4.7 Summary and Recommendations B-81
B4.8 Literature Cited and References B-84
B5.0 Plankton: Biomass, Productivity and Community Structure/Function B-89
B5.1 Rationale B-89
B5.2 Monitoring Design Considerations B-89
B5.3 Existing Analytical Methods B-93
B5.4 QA/QC Considerations r B-95
B5.5 Statistical Design Considerations B-96
B5.6 Use of Data B-96
B5.7 Summary and Recommendations B-97
B5.8 Literature Cited and References B-99
B6.0 Aquatic Vegetation B-109
B6.1 Rationale B-110
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B6.2 Monitoring Design Considerations B-110
B6.3 Existing Analytical Methods B-lll
B6.4 QA/QC Considerations B-117
B6.5 Statistical Design Considerations , B-l 18
B6.6 Use of Data B-118
B6.7 Summary and Recommendations B-l 19
B6.8 Literature Cited and References B-121
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B7.0 Benthlc Infauna Community Structure B-127
B7.1 Rationale B-127
B7.2 Monitoring Design Considerations B-127
B7.3 Existing Analytical Methods B-136
B7.4 QA/QC Considerations B-144
B7.5 Statistical Design Considerations B-145
B7.6 Use of Data B-146
B7.7 Summary and Recommendations B-147
B7.8 Literature Cited and References '. B-149
B8.0 Fish Community Structure B-155
B8.1 Rationale B-155
B8.2 Monitoring Design Considerations B-155
B8.3 Analytical Methods Considerations B-157
B8.4 QA/QC Considerations B-162
B8.5 Statistical Design Considerations B-163
B8.6 Use of Data B-163
B8.7 Summary and Recommendations B-164
B8.8 Literature Cited and References B-165
B9.0 Fish and Shellfish Pathobiology B-169
B9.1 Rationale B-169
B9.2 Monitoring Design Considerations B-171
B9.3 Existing Analytical Methods B-174
B9.4 QA/QC Considerations B-179
B9.5 Statistical Design Considerations B-180
B9.6 Use of Data B-181
B9.7 Summary and Recommendations B-184
B9.8 Literature Cited and References B-188
B10.0 Bioaccumulation B-199
B10.1 Rationale B-199
B10.2 Monitoring Design Considerations B-200
B10.3 Existing Analytical Methods B-209
B10.4 QA/QC Considerations B-211
B10.5 Statistical Design Considerations B-215
B10.6 Use of Data B-215
B10.7 Summary and Recommendations B-215
B10.8 Literature Cited and References B-220
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Bll.O Bacterial and Viral Pathogens B-225
Bll.l Rationale B-225
B11.2 Monitoring Design Considerations „ B-226
B11.3 Analytical Methods Considerations B-227
B11.4 QA/QC Considerations B-234
B11.5 Statistical Design Considerations B-234
B11.6 Data Use B-234
B11.7 Summary and Recommendations B-235
B11.8 Literature Cited and References B-237
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xiii
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Tables
Table Page
1-1. Elements of Systems Approach « 8
2-1. Puget Sound Ambient Monitoring Program Design Specifications for
Sediment Chemistry Sampling 21
3-1. Examples of Monitoring Program Objectives and Associated Questions '. 26
3-2. References to Basic Monitoring Design and Statistical Texts 29
Al-1. Alternatives for Improving Puget Sound Monitoring A-6
Al-2. MMC Activities Required by PSWQA in Developing PSAMP A-8
Al-3. PSAMP Design and 1989-90 Activities A-12
Al-4. PSAMP Task Assignments by Agency A-15
A2-1. Seasonal Kendall Tau Result A-37
B-l. Sampling Methods Described , B-4
B-2. Format of Methods Section B-5
B-3. Technical Support and Guidance Documents B-6
B-4. Monitoring Topics Included in Puget Sound Estuary Program Protocols B-8
B-5. Key Topics to be Addressed in Estuary Program Quality Assurance Plans B-9
B-6. QC Sample Types B-ll
Bl-1. List of Methods and Equipment B-20
Bl-2. Recommended Sample Preservation and Storage Requirements B-25
Bl-3. Recommended Analytical Methods B-26
B2-1. List of Existing Analytical Techniques B-39
B2-2. Sample Preservation and Storage Parameters B-43
B2-3. Definitions for Selected Limits of Detection B-45
B3-1. Summary of Bottom Sampling Equipment B-54
B3-2. Sediment Grain Size: Withdrawal Times for Pipet Analysis as a Function of
Particle Size and Water Temperature B-60
B4-1. List of Existing Analytical Techniques B-72
B4-2. Summary of Sample Collection and Preparation QA/QC Requirements for Organic Compounds... B-76
B4-4. Sampling Containers, Preservation Requirements, and Holding Times for Sediment Samples B-77
B4-5. Summary of Quality Control Sample B-80
B4-6. Summary of Warning and Control Limits for Quality Control Sample B-81
B6-1. List of Terms B-109
B6-2. List of Analytical Methods B-112
B7-1. Summary of Bottom Sampling Equipment B-129
B7-2. Biological Indices B-137
B8-1. Biological Indices B-159
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Table
B9-1.
B9-2.
B10-1.
B10-2.
B10-3.
B10-4.
B10-5.
B10-6.
B10-7.
B10-8.
Bll-1.
Bll-2.
Page
List of Pathobiological Terms B-170
Highest Ranking Candidate Fishes for Use as Pathobiology Monitoring Species B-172
List of Terms • • B'199
Highest Ranking Candidate Fishes for Use as Bioaccumulation Monitoring Species B-202
Recommended Large Macroinvertebrate Species for Bioaccumulation Monitoring B-204
List of Existing Analytical Techniques • B-211
Sample Preservation and Storage Paramters B-212
Summary of Sample Collection and Preparation QA/QC Requirements for
Organic Compounds • B-213
Summary of Quality Control Sample B-216
Summary of Warning and Control Limits for Quality Control Sample B-217
Microorganisms Responsible for Causing Adverse Human Health Effects B-228
Laboratory Procedures for Bacterial Indicators B-231
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XV
Figure
• w.
Figures
Page
1-1. Monitoring Program Design 9
2-1. Impacts on the Marine Environment of the Southern California Bight 19
4-1. Description of Various Sampling Methods 33
5-1. Hypothesis Testing: Possible Circumstances and Test Outcomes 46
5-2. Minimum Detectable Difference vs. Number of Replicates for Fixed Set of
Design Parameters 48
5-3. Power y^. Mniimum Detectable Difference 49
7-1. Sample Coyer. pfPuget Sound Notes 57
7-2. Sample Cover of Chesapeake Bay Barometer 58
7-3. Sample Covers of Santa Monica Bay Restoration Project Reports 58
A1 -1. Events Leading to the Development and Implementation of the Puget Sound
Ambient Monitoring Program A-4
Al-2. Area Included in Puget Sound Ambient Monitoring Program A-ll
A2-1. Station Locations Included in Data Subset of the Chesapeake Bay
Ambient Monitoring Program A-31
A2-2. Example Box Plot... A-33
A2-3. Time Series for N as NH3 for Surface Stations A-34
A2-4. Time Series for Dissolved O2 for Bottom Stations A-34
A2-5. Box Plot for N as NH3 for Surface Stations A-35
A2-6. Box Plot for Dissolved O2 for Bottom Stations A-35
A2-7. Power Analysis forN as NH3 for Surface Stations A-39
A2-8. Power Analysis for Dissolved O2 for Bottom Stations A-39
B4-1. Examples of Acceptable and Unacceptable Samples B-75
B7-1. Generalized SAB Diagram of Changes Along a Gradient of Organic Enrichment B-139
B7-2. Diagram of Changes in Fauna and Sediment Structure Along a Gradient of Organic Enrichment...B-141
B7-3. Examples of Acceptable and Unacceptable Samples B-144
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1.0 Introduction
The Clean Water Act as amended by the Water Quality Act of 1987 establishes the
National Estuary Program (NEP) to promote long term planning and management in
nationally significant estuaries threatened by pollution, development, or overuse.
Section 320 of the Clean Water Act describes the establishment of a management
conference in each estuary to develop a Comprehensive Conservation and Manage-
ment Plan. It also establishes requirements to monitor the effectiveness of actions
taken pursuant to the plan.
This document provides guidance on the design, implementation, and evaluation of the
required monitoring programs. This document should also be of use to others develop-
ing monitoring programs. The intended audience is the members of the management
conference (i.e., the Management Committee, Scientific and Technical Advisory
Committee, and the Citizens Advisory Committee) and the program coordinators and
scientific staff of the individual estuary programs. The purpose of this document is to
identify the steps involved in developing and implementing estuarine monitoring
programs. Given the large number of participants in the management conference and
the diversity of technical backgrounds, it is also intended to provide a technical basis
for discussions on the development of monitoring program objectives, the selection of
monitoring program components, and the allocation of sampling effort. Some of the
issues addressed in this document are:
• What is the role of monitoring in the estuary programs?
• Why are monitoring programs necessary, and what are examples of goals
and objectives?
• What criteria should be used to select components of the monitoring
program, and what should drive the decision-making process?
• What is the importance of historical data, and how can ongoing monitoring
programs be incorporated into the estuarine monitoring program?
• What is the relationship between estuary characterization and the monitor-
ing program?
The key technical and programmatic aspects associated with each of these issues are
described. This document also includes several examples. Two case studies are used
to provide examples from existing estuarine monitoring programs. While these case
studies focus on unique sets of problems and questions from individual programs, they
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provide illustrations of the process of monitoring program design, implementation, and
evaluation. The lessons learned are of general applicability even though the political
and institutional setting may be different from other estuaries.
The first case study from the Puget Sound Estuary Program (Appendix A 1.0) provides
an example of the process of developing an effective monitoring strategy. Key issues
addressed include problem and goal definition, the process of monitoring program
design and implementation, and options for funding estuary monitoring programs. The
first case study also demonstrates how ongoing monitoring studies can be coordinated
to develop a comprehensive basin-wide monitoring program. The second case study
(Appendix A2.0) provides a detailed example of the application of methods for
determining the effectiveness and feasibility of monitoring efforts in the Chesapeake
Bay Program. Examples from other environmental monitoring programs are also
described.
Appendix B of this document describes existing methods that may be used to monitor
the effectiveness of management actions in the estuary. The National Estuary Program
docs not impose national requirements for standard methods. However, emphasis is
placed on the importance of using standardized monitoring protocols within each
csiuary and developing performance-based criteria to evaluate the comparability of
analytical methods. One long-term goal of the NEP is to develop a standardized set of
key variables that, if measured in all NEP programs, will provide comparative data on
nationally significant estuaries.
Funding is typically drawn from federal, state, and local sources for National Estuary
Program monitoring activities. Therefore, this document discusses the integration of
existing monitoring efforts into the estuary monitoring program. It is also essential
that National Estuary Program monitoring activities be coordinated with existing
federal agency status and trends monitoring programs such as EPA's Ecosystem
Monitoring and Assessment Program (EMAP), the National Oceanic and Atmospheric
Administration's (NOAA) Status and Trends Program, and the U.S. Geological
Survey's National Water Quality Assessment Program. If individual estuary monitor-
ing programs will be measuring the same parameters as these federal programs, and
the protocols in use provide adequate data, estuary programs should work to ensure
compatibility of their data with the existing federal programs.
Data management, effective data analysis, and the communication of monitoring
program results to a wide range of audiences at several technical levels are also
essential to the success of the estuary program. This document discusses data manage-
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ment strategies and provides several examples of ongoing efforts to disseminate
information developed by the National Estuary Program.
1.1
Background
National Estuary Program
The purpose of the NEP is to identify estuaries of national significance and to promote
the preparation of comprehensive management plans to ensure their ecological integ-
rity. The process of identifying nationally significant estuaries and nominating estuar-
ies to become part of the NEP is described in A Primer for Establishing and Managing
Estuary Projects (U.S. EPA, 1989a) and Final Guidance on the Contents of a
Governor's Nomination (U.S. EPA, 1990a). Following selection of an estuary to be
included in the National Estuary Program, a management conference is convened by
the EPA Administrator. The management conference has the responsibility to imple-
ment a four-phased program:
• Phase I - The Planning Initiative. The planning phase is intended to build
the management framework for identifying and solving problems and
defining the steps in the decision making process.
• Phase II - Characterization and Problem Definition. The goal of estuary
characterization is to gather and summarize the existing knowledge con-
cerning the state of the estuary as well as the physical, chemical, and
biological factors controlling spatial and temporal changes. The character-
ization process focuses on identifying existing and potential problems and
exploring probable causes of such problems. Such cause and effect
linkages between human activities and environmental changes provide the
public and decision makers with the information necessary to develop
priorities, set management strategies, and devise mitigating measures.
However, information gaps do exist. Such information gaps can be filled
by additional sampling and a subsequent monitoring program. The charac-
terization process is described in a separate EPA guidance document.
• Phase HI - Development of a Comprehensive Conservation and Manage-
ment Plan (CCMP). The CCMP is a major product of the estuary program.
It is developed by the management conference to summarize findings,
identify and prioritize problems, determine environmental quality goals
and objectives, identify action plans and compliance schedules for pollu-
tion control and resource management, and to ensure that designated uses
of the estuary are protected.
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• Phase IV - CCMP Implementation. When the final CCMP is submitted
for approval to the administrator, the management conference and the
governor establish a committee to coordinate the implementation of the
CCMP. The development of a monitoring program to evaluate the effec-
tiveness of actions specified in the CCMP is a required task of the manage-
ment conference.
The four phases of the NEP should not be viewed as sequential steps, where one phase
cannot be initiated before another phase is completed. On the contrary, as the NEP has
evolved, EPA has encouraged management conferences to proceed with the four
phases simultaneously as often as possible. For example, early results of characteriza-
tion (Phase II) may indicate obvious management actions that can be taken (Phase IV)
prior to completion of the CCMP (Phase III), In these cases, implementation actions
should proceed. In fact, EPA will base the selection of any new estuaries on an ability
to streamline the NEP phases, focusing on estuaries that have a significant amount of
the problem characterization done, that have a successful management framework that
functions similarly to an NEP management conference already established, and that
have clear commitments from key state and local agencies to participate in and support
the NEP process.
Environmental sampling is required in Phases II and IV of the estuary programs. The
studies conducted in Phase n are focused on filling identified data gaps, providing
baseline data on point and nonpoint loadings of pollutants, and developing estimates of
the degree of spatial and temporal variability. Sampling is conducted in Phase IV of
the estuary programs as part of a long term environmental monitoring strategy.
Environmental monitoring that is conducted during Phase IV is considered to be
different from the sampling that is conducted during Phase II. Monitoring involves
repeated sampling over time. For example, short term sampling may be conducted in
Phase II to collect specific information on the concentration, distribution, and variabil-
ity of chemical contaminants in sediments. The goal of the corresponding sampling
that is conducted during Phase IV is to evaluate trends in monitored variables and to
link the observed patterns to specific management actions. A second distinction
between Phase n sampling and Phase IV monitoring is that comprehensive environ-
mental monitoring programs conducted during Phase IV will require the integration of
information from several concurrent sampling efforts. Since environmental sampling
is costly and resources will be limited, there will be a need to evaluate the efficacy of
different monitoring program components and to allocate sampling efforts accord-
ingly. It will also be necessary to carefully plan and coordinate monitoring efforts
among individual monitoring components and other preexisting monitoring programs
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to determine interactions and streamline monitoring efforts. A third distinction
between environmental sampling and monitoring is the need to periodically analyze
the monitoring program results and modify the level of sampling effort to maximize
program effectiveness.
NEP Monitoring Program Goals
EPA is developing a programmatic monitoring system for NEP participants to use in
tracking the progress being made in protecting NEP estuaries. While primarily aimed
at tracking implementation of actions recommended by the management conferences
in their CCMPs, the system will be helpful in the assessment of the NEP in its entirety.
This programmatic monitoring system will:
• Assist estuary program managers to improve their programs by identifying
current and emerging programs;
• Provide accountability to elected officials and the public relating to the
progress toward estuary protection;
• Help identify the programs and projects that are working well; and
• Provide a framework for assessing the NEP as a whole.
This programmatic monitoring system will identify estuary management performance
indicators that will track, on a regular basis, the status and progress of action-oriented
efforts. Much of the input to this programmatic monitoring system will come from
information generated by NEP environmental monitoring programs implemented
under Phase IV of the NEP, as well as other environmental monitoring programs.
The two primary goals of the monitoring programs implemented in Phase IV by the
management conferences are:
• To measure the effectiveness of management actions and programs imple-
mented under the CCMP and
• To provide essential information that can be used to redirect and refocus
the management plan.
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These monitoring programs are an essential part of the management program review
and evaluation process for each estuary, and they will be conducted throughout the
implementation of the CCMP.
This document describes the steps involved in designing an environmental monitoring
program to meet these two interrelated goals. For example, the goals of the Puget
Sound Ambient Monitoring Program (described in Case Study 1) include the charac-
terization and interpretation of spatial and temporal patterns of conditions in Puget
Sound, development of a permanent record of significant natural and human-caused
changes in key environmental indicators, and the support of research activities through
the availability of consistent, scientifically valid data. Such research will lead to a
better understanding of the state of the estuary in question and, in turn, provide infor-
mation for making better resource management decisions.
Monitoring Design Issues
Marine monitoring is the continued, systematic time-series observation of
predetermined pollutants or pertinent components of the ecosystem over a
period of time sufficient to determine (1) the existing level, (2) trends, and
(3) natural variations of measured components (NQAA, 1979).
Recent reports by the National Research Council (NRC, 1990a and 1990b).evaluated
current marine monitoring programs and practices, identified needed improvements in
monitoring strategies, and made a series of recommendations to improve the useful-
ness of monitoring information.
The extensive NRC review of monitoring practices found numerous inadequacies in
the design and use of monitoring data. Three broad problem areas were identified:
• Monitoring programs are often poorly designed because of failure to
clearly define monitoring objectives and to apply available design tools.
Some of the identified problems could be attributed to the inherent diffi-
culty of separating the effects of human activities from natural variability.
• There is a lack of communication and coordination among the regulatory,
scientific, and management entities sponsoring or conducting monitoring
programs. Specific concerns include the inflexibility of regulatory require-
ments that limit opportunities to adapt programs to new needs and regional
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objectives. The need to adopt standardized sampling and quality assurance
procedures to ensure data comparability has also been identified.
• The results of monitoring programs are not presented in a form that is
useful in developing broad public policy or evaluating specific control
strategies. It is essential to link data management strategies and data
analysis methods to the objectives of the monitoring effort. It is also
necessary to devise a plan for effectively communicating monitoring
results to the identified audience.
Additionally, the NRC study found that poorly designed monitoring programs and the
lack of communication and coordination among programs has often resulted in:
• Limitation of water quality and public health monitoring to areas adjacent
to point-source outfalls and other known or potential problem areas, and to
the sampling and analysis of conventional water quality and public health
parameters
• Living resource monitoring programs restricted to assessment of a few
higher-trophic levels of commercially or recreationally important species,
without consideration of key prey species upon which these species depend
• Incomplete monitoring of living resources and habitats key to the determi-
nation of potential declines in living resources and water quality (i.e., near
shore habitats, estuarine wetlands, plankton communities)
1.2
Recommended
Monitoring Design
Procedures
A wide range of guidance on the design of environmental monitoring programs is
available in the literature. Important contributions have been made that address the
principles and options for designing monitoring networks (Green, 1979; Segar and
Stamman, 1986), the development of monitoring objectives (Beanlands and Duinker,
1983), the appropriateness of monitoring variables (Bilyard, 1987), and the application
of statistical methods in the design process (Ferraro et al, 1989). The recent NRC
study (NRC, 1990a) also provides comprehensive review of the steps involved in the
design of marine environmental monitoring programs.
Outlined below is a,systems approach to the design of the required NEP monitoring
programs that incorporates existing information and that will ensure the collection,
analysis and reporting of adequate information to meet the goals of individual estuary
programs.
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8
TABLE 1-1. ELEMENTS GF SYSTEMS APPROACH
Define the Objective
Establish Information Needs
Establish the Objectives of
Individual Program Components
Evaluation of Trade-Offs
Feedback to Initial Design Step -
The overall objective of the design process Is
stated in a succinct manner.
Information requirements to meet the
objectives are established.
The objectives of all possible monitoring
program components and performance
criteria are established*
The combination of monitoring components
- that best meet the overall objectives is
selected,
Modifications to the system's design are made
to improve the product's performance.
A systems design approach places emphasis on the optimum design of, the overall
monitoring program. The essential elements (shown in Table 1-1) are the assessment
of trade-offs between individual aspects of the monitoring program and the use of
feedback mechanisms to modify individual monitoring program components based on
periodic assessments of overall program performance. This approach is well suited for
design problems that involve complex, highly variable systems, such as estuaries, and
that involve a large number of investigators that must interact as a group to produce
the product.
The plan for implementing this approach to designing the monitoring program is
Shown in Figure 1-1. It involves the specification of monitoring objectives, the
completion of a series of steps to translate these objectives into clearly defined moni-
toring activities, and the use of feedback mechanisms to refine the objectives and
adjust the sampling effort.
The overall objective of monitoring undertaken during Phase IV of the estuary pro-
grams will be to measure the effectiveness of management actions implemented as part
Of the CCMP. Meeting this broad program objective will require the specification of
several individual, highly interrelated monitoring objectives. Examples of individual
monitoring objectives (shown in Figure 1-1 as Objectives 1 through n) include: to
determine the response of key water quality variables to management actions; to
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Public
Concerns
Develop
Monitoring Objectives,
Performance Criteria
DEVELOP / REFINE MONITORING OBJECTIVES
Establish Testable
Hypotheses and Select
Statistical Methods
Select Analytical Methods
and Alternative
Sampling Designs
Evaluate Expected
Monitoring Program
Performance
Is
Monitoring
Program
Performance
Adequate?
EVALUATE / ASSESS PROGRAM PERFORMANCE
COMMUNICATE
MONITORING
RESULTS/REDIRECT
MANAGEMENT
PROGRAM
Design and
Implement Data
Management Plan
Figure 1-1. Monitoring
program design.
determine trends in sediment contaminant concentrations; and, to evaluate the persis-
tence of PCBs in the tissue of recreational and commercial fish.
Each monitoring objective represents a separate component of the overall estuary
monitoring program. The individual steps involved in designing each component of
the monitoring program are shown on the right-hand side of Figure 1-1:
• Step 1. Develop Monitoring Objectives and Performance Criteria. Clear
objectives and corresponding performance criteria must be developed for
each component of the monitoring program. Performance criteria specify
the level of change or trend that the monitoring program must be able to
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10
detect. For example, in the Chesapeake Bay Program, a target of 40%
reduction in nutrient loads to the bay was established based on modeling
results. One of the predicted benefits of nutrient reduction is the allevia-
tion of anoxic conditions in the bay and an increase in the minimum
dissolved oxygen to approximately 1.5 mg/l. Therefore, the corresponding
objective and performance criterion for the water quality component of the
monitoring program would be to detect a long term change in dissolved
oxygen concentration in the bottom waters of the bay equal to 1.5 mg/t.
Step 2. Establish Testable Hypotheses and Select Statistical Methods. The
study objectives must be translated into statistically testable hypotheses;
for example, no trend exists in measured values of dissolved oxygen
concentration. Establishing testable hypotheses ensures that the results of
the monitoring program will be unambiguous and that the objectives of the
program can be met The establishment of testable hypotheses also guides
the development of statistical strategies for determining sample locations
and times as well as the selection of statistical tests that will be used to
analyze the resulting data.
Step 3. Select Analytical Methods and Alternative Sampling Designs.
Detailed specifications for each monitoring variable (measurable endpoint)
of the monitoring program must be developed. These include: field
sampling and laboratory procedures, and QA/QC procedures. Addition-
ally, alternative sampling designs that specify the number and location of
stations must be devised for input to the next step in the design process.
Step 4. Evaluate Expected Monitoring Program Performance. It is essential
to evaluate the expected performance of the initial sampling design to
determine the minimum difference that can be detected over time or between
locations. Without this evaluation there is a risk of collecting and analyzing
too few samples to detect statistically significant temporal or spatial trends
or of analyzing an excessive number of samples (with associated high costs).
As indicated by the feedback loop shown in Figure 1-1, the results of this
evaluation are used to identify modifications to the initial design in order to
increase monitoring program effectiveness. Information from this evalua-
tion will also be used to assess the ability of monitoring components to
provide information used to modify the management plan.
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11
• Step 5. Design and Implement Data Management Plan. The development
of a data management system is an essential task that is often overlooked in
the design of monitoring programs. The data management system should
be operational prior to implementation of the monitoring program. Data
analysis methods and a timetable for analyzing the data, assessing CCMP
implementation progress and monitoring program performance, and
reporting program results should also be specified. The results of the
performance assessment are used to refine program objectives and modify
individual study elements to satisfy these objectives.
These individual steps in the monitoring design process shown in Figure 1-1 are
described in Sections 2 through 6 of this document.
Peer review of the monitoring program is recommended to evaluate and assess pro-
gram design. Critical review by technical experts without a vested interest in the
estuary program will ensure that the monitoring objectives are meaningful and that the
monitoring strategy adequately addresses these objectives with the most appropriate
methods. This review should take place after the initial development of specific
objectives and performance criteria and as part of periodic reviews that are conducted
to evaluate the success of the monitoring program.
Overall program performance must be assessed at periodic intervals, and the results
should be used to refine monitoring program objectives and methodologies. The
original monitoring design must remain open to modification. The monitoring pro-
gram should take advantage of new information and innovative sampling approaches
as they are developed, and the link between modeling and monitoring efforts should be
fully exploited. The results of the monitoring program should be used to refine and
modify conceptual and mathematical models of the system, and modeling results
should be used to guide changes in individual monitoring program components,
variables monitored, the frequency of sampling, and overall monitoring strategies. It is
essential that new information, from both independent research and the monitoring
program results, is integrated into the monitoring program.
1.3
Monitoring
Program
Management
There are a number of management issues related to the design, implementation, and
maintenance of NEP monitoring programs that must be addressed early by the man-
agement conference. These include setting the timetable for the design and implemen-
tation process, assigning responsibilities for coordinating the design effort, and plan-
ning for the long term success of the monitoring program.
-------
Timetable for the Design and Implementation of the Monitoring Program
The design and implementation of the Puget Sound Ambient Monitoring Program
(PSAMP), described in Case Study 1, required four years of uninterrupted effort. The
development of the monitoring program was a consensus building effort among
numerous agencies and organizations, and several iterations were required to finalize
the initial design plans. In addition to the decisions regarding the basic sampling
strategy, agreement was required on numerous interrelated issues, such as sampling
protocols, appropriate quality assurance/quality control methodology, and the selection
of an information management system. • :
Planning for the monitoring program should be initiated during the first year of the
management conference. Milestones for the monitoring program effort should be
clearly stated in the State-EPA Management Conference Agreement (a three to five
year action plan for CCMP completion that is negotiated shortly after a management
conference is convened). Development of the monitoring program should be given a
high priority by the management conference after it is convened. It is important to
begin early planning of the monitoring program to ensure that it is in place at the time
the CCMP is implemented. A detailed monitoring program plan must accompany the
CCMP that is submitted to the EPA Administrator for approval. The monitoring plan
must contain the following elements that are described in this document:
• Definition of program objectives and performance criteria (parameter
values needed to guide management decisions)
• Identification of testable hypotheses
• Detailed specifications for each monitoring variable, including sampling
locations and frequency, field sampling procedures, field and laboratory
analytical procedures, quality assurance arid control procedures
• Specification of the data management system and statistical tests that will
be used to analyze the monitoring data
• Description of the expected performance of the initial sampling design
(i.e., the minimum difference that can be detected in measured variables
over time and between locations) ,
• Plan and timetable for analyzing data and assessing program performance
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13
Management Tasks for Developing the Monitoring Program
The first management task in developing the monitoring program should be the
establishment of an organization or committee, such as a monitoring subcommittee of
the Scientific and Technical Advisory Committee (STAC), to develop the monitoring
program. The STAC is one of three committees that make up the NEP management
conference. The other two committees are the management committee and the
Citizen's Advisory Committee. Membership in the monitoring subcommittee should
not be limited to STAC members but should include representatives from federal, state
and local agencies, universities, industry, environmental groups and others currently
conducting monitoring or planning monitoring programs within the estuary or sur-
rounding waters. The subcommittee should be charged with the following tasks:
• Define the goals and objectives of the monitoring program.
• Propose an initial design that includes recommendations for sampling and
analytical protocols, data management system specifications, quality
assurance guidelines, data reporting requirements and cost estimates.
» Develop the final monitoring design using workshops and other mecha-
nisms to solicit comments and suggestions from the public and scientific
community.
• Coordinate the activities of the numerous interested and participating
agencies and develop interagency agreements that will promote the moni-
toring effort.
• Identify funding mechanisms and opportunities to contain costs.
As discussed in Section 1.2, the process of developing a comprehensive monitoring
program must begin with a clear statement of the objectives. The explicit statement of
objectives, and options for obtaining these objectives, is necessary as a starting point for
describing the problem areas in the estuary and developing the consensus among inter-
ested agencies and other parties that is essential to the success of the monitoring effort.
The primary goals of the monitoring program will be to measure the success of the
CCMP and to provide information that can be used to redirect and refocus the manage-
ment. However, it will also be the responsibility of this subcommittee to develop
secondary goals and program objectives that will be used to focus the sampling effort.
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14
These objectives could include: continued characterization of spatial and temporal
patterns of change in water quality, sediment and biological resources of the estuary;
development of a permanent record of significant natural and human-caused changes
in environmental indicators in the estuary over time, and support for research activities
through the availability of consistent, scientifically valid data. The process of develop-
ing these objectives is described in Section 2.0.
There are several options for initiating the design process. The PS AMP began with an
initial monitoring design developed by consultants to EPA Region 10. The initial
design included goals and objectives, plans for operation of the program, and methods
for sampling, analysis, and reporting the data. The first draft of the monitoring pro-
gram design was refined by EPA Region 10 through a process involving meetings and
discussions among managers and scientists working in the Puget Sound region. The
Puget Sound Monitoring Management Committee then proceeded to review, modify
and refine the proposed monitoring design. This resulted in the development of a
revised draft that was released for public and further scientific review. The Santa
Monica Bay Restoration Project began the development of the monitoring program
with an "Assessment of Monitoring and Data Management Needs Workshop" to
develop and evaluate monitoring options. In preparation for the workshop, back-
ground information was compiled and used to develop a "straw man" to guide work-
shop discussions. In both the Puget Sound and the Santa Monica programs, public and
scientific workshops were held to discuss the proposed monitoring options and to build
the required consensus among participating entities.
The next task following the adoption of the monitoring program design is to ensure
that it is implemented as planned. The implementation of a regional or basin-wide
monitoring program requires the development of commitments and the coordination of
activities among the participating agencies. This was achieved in the Puget Sound
Ambient Monitoring Program by negotiating memoranda of agreement (MOA) with
each of the participating agencies. The MO As specified commitments that each
agency carry out its responsibilities identified in the monitoring program design, and
that each agency maintain monitoring program funding levels and staff support for the
monitoring program committees. As described in Case Study 1, the negotiating of the
MO As was essential to the success of the PS AMP, but the complexity of the process
was underestimated. It is recommended that the coordination efforts begin as soon as
possible in the design process.
It is also important that the monitoring subcommittee begin to identify, early in the
process of monitoring program development, the costs associated with existing moni-
toring in the basin, the costs of additional monitoring that will be needed, potential
-------
15
sources of additional funds to support the monitoring program, and appropriate mecha-
nisms to fund the monitoring program. Costs of the Puget Sound Ambient Monitoring
Program were calculated by a technical costing subcommittee of the monitoring
management committee with the assistance of a technical consultant.
The estimates provided by this subcommittee demonstrate that the costs of comprehen-
sive monitoring programs can be substantial. In addition to the $200,000 in staff and
consultant time required to develop the monitoring program design, the calculated
costs of full implementation of the PSAMP are approximately $3.2 million per year (in
1987 dollars). The initial sampling program was reduced in scope due to resource
constraints, and costs for the program have been $250,000 to $350,000 over the first
two years. The projected costs of implementing the monitoring plan submitted as part
of the Buzzards Bay CCMP are $750,000 per year. This includes $200,000 funded by
the estuary program to supplement ongoing state and citizen monitoring programs.
The costs of the comprehensive water quality monitoring in the mainstem of the
Chesapeake Bay are on the order of $900,000 annually. This monitoring includes the
sampling of 20 variables at 49 stations, 20 times. In addition to mainstem sampling,
the states surrounding the bay have undertaken their own extensive sampling programs
in the bay's tributaries.
The substantial costs of these programs require equally substantial efforts to secure
long term funding. Most of the costs of monitoring should be borne by the state and
local agencies that will complete specific monitoring tasks. Therefore, the best
opportunity for securing the required funding is indirectly by incorporating existing
federal, state and local monitoring programs into the overall design. The critical step
will be to motivate the participating agencies to make modifications to existing moni-
toring efforts to meet the goals of the estuary program. The best way to achieve this
objective is to involve these agencies in the estuary program and the monitoring design
process as early as possible and to demonstrate the benefits of a regional / basin-wide
monitoring program to the scientific and regulatory community. If state and local
funds are not immediately available for full implementation of the monitoring pro-
gram, a plan for phased implementation can be developed through a priority setting
process involving the monitoring management committee and other technical experts.
Section 4.0 of this document discusses the incorporation of individual components of
ongoing local monitoring programs into the National Estuary Program monitoring
efforts. Opportunities for taking advantage of monitoring programs of national scope,
such as EPA's Environmental Monitoring and Assessment Program (EMAP) and
NOAA's Status and Trends Program, as well as locally coordinated citizen monitoring
programs, are also discussed.
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16
(i 'li,,
Financing Marine and Estuarine Programs: A Guide to Resources (U.S. EPA, 1988a)
provides guidance for obtaining direct financing of marine and estuarine monitoring
programs. This document provides a primer on basic financing concepts and explains
the initiatives needed to begin financial planning for long term resource management
activities. Case studies are also included that provide examples of local financial efforts.
Planning for the Long Term Success of the Monitoring Program
A structure for managing the monitoring program must be recommended to the
management conference by the monitoring subcommittee or its equivalent. A lead
coordinating agency or organization must be identified and given the responsibility for
implementing the monitoring design as planned, maintaining the monitoring effort,
and reviewing the monitoring program results. The lead agency or organization should
provide a full-time manager/coordinator and a staff that is responsible for keeping all
the implementing agencies and other participants informed of progress, resolving
disputes, carrying out technical and administrative duties, and reporting the results of
the program.
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17
2.0 Develop Monitoring Objectives and
Performance Criteria
The overall objective of the estuary monitoring program is to assist in determining the
effectiveness of the implementation of the CCMP. However, this overall objective
may encompass several specific monitoring objectives. The identification of these
specific objectives begins during Estuary Characterization. The characterization
process identifies public concerns and formulates a series of corresponding
management issues. During characterization, conceptual and predictive models are
developed and research results are evaluated to provide a basic understanding of
important physical, chemical and biological processes in the estuary. This information
is used in the design of the monitoring program to specify a set of variables and
ecological processes that can be used to detect changes in the estuary in response to
management actions.
Regardless of the scope of the proposed monitoring program, it is essential to develop
explicit statements of the monitoring objectives as well as to establish performance
criteria with which to measure monitoring program success.
MONITORING OBJECTIVES,
PERFORMANCE CRITERIA
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18
The development of the monitoring objectives is the culmination of the estuary
characterization and the preparation of the CCMP. The characterization process is
described in a separate technical guidance document (U.S. EPA, 1992). It involves the
identification of public concerns and potential water quality, biological and public
health problems in the estuary. A unique approach for identifying these concerns and
ecological problems is based on previous work summarized in the National Research
Council's Managing Troubled Waters (Clark, 1986; NRC, 1990a and 1990b). The key
to this approach is the construction of the matrix, shown in Figure 2-1, which identifies
Valued Ecosystem Components (VECs) and sources of perturbation. This matrix also
summarizes the understanding of the relative importance of each source of
perturbation and the degree of scientific certainty associated with knowledge about
each impact. Each cell in the matrix summarizes the effects of each perturbation on a
single ecosystem component. For example, the information summarized in Figure 2-1
indicates that wastewater outfalls are a controlling factor of soft-bottom benthos
communities and that there is a moderate degree of certainty regarding the scientific
understanding of the effects of these outfalls in the Southern California Bight. The
effects of each identified source of perturbation (e.g., storm water runoff) on all
identified resources (VECs) are summarized along a single row. Similarly, each
column summarizes the existing knowledge of the impacts on a single resource caused
by the complete range of identified sources of perturbation.
This matrix summarizes existing information on the resources of the estuary and
potential impacts in an easily accessible manner. The process of developing this
matrix also provides an effective tool for building consensus among the wide range of
interested parties in the estuary program on the relative priority of monitoring objec-
tives and the different components of proposed monitoring programs. Bernstein et al.
(1991) describe an Integrated Assessment methodology based on the work by Clark
(1986) that guides the selection of monitoring objectives based on the relative impor-
tance of the identified resources, the understanding of the underlying controlling
processes, and the ability to detect changes in monitoring variables.
Simple conceptual and predictive models developed in the characterization process
may also be used to summarize the physical, chemical, geological and biological status
of the estuary and identify the factors controlling spatial and temporal changes.
Finally, it is necessary to quantify the identified ecological relationships. Existing data
should be analyzed to evaluate the strength and direction of identified relationships
and to determine the magnitude of uncertainty associated with the existing
information. The products of the characterization process should include the
identification of the primary management issues. These management issues are used
2.1
Monitoring
Program
Objectives
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19
h', " *
V ,**•.. ' - •*• "•>
Figure 2-1.
Impacts on the marine
environment of the
Southern California Bight
(Bernstein ej. aL., 1991).
VALUED
ECOSYSTEM
\COMPONENTS
SOURCES OF
PERTURBATION
Storms
El Ninos
California Current
Upwelling
Blooms/Invasions
Ecol. Interactions
Power Plants
Wastewater Outfalls
Dredging
Rivers/Storm Runoff
Commercial Fishing
Sport Fishing
Habitat Loss/Mod.
Oil Spills
All
Intertidal
4>
T1
<£D>
^P
^
Ji\
^iil'
m\
N||y
Phytoplankton
^
TJ^
^
^P
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^
Zooplankton
©
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™
^p,
Soft Bottom Benthos
rt
14^
s\
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IP
i
^
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i
Hard Bottom Benthos
O
^1
1
Q.
2
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\)
fT\
(I)
(^
NlP
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^
|p
W
Wetlands & Estuaries
^
^3
^
1
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^y
16
ib
Commercial Shellfish
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^
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^
^
W
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Net effect of all sources on each component
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feet of each source on all components
^
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KEY
Potential Importance
Controlling Major Moderate Some
v37 vb '^E? ^1
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High Moderate
Low
Iffl
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20
••>»*,
for the development of the CCMP. The CCMP sets environmental quality goals and
management objectives for the estuary and specifies actions for achieving these
objectives. The monitoring program is designed to verify the predicted results and
evaluate the effectiveness of the CCMP implementation, and to recommend corrective
actions. When insufficient information is available for estuary characterization and/or
CCMP development, a monitoring program may also be designed to fill the
information gaps.
Case Study 1 (Appendix Al.O) describes the process that led to the development of the
goals of the Puget Sound Ambient Monitoring Program. The essential steps were:
• The development of the Puget Sound Water Quality Management Plan - a
comprehensive management plan for Puget Sound and its related water-
ways. In developing this plan, nine issue papers were prepared. One of
these, Comprehensive Monitoring of Puget Sound (PSWQA, 1986),
reviewed existing monitoring programs and described the process of
developing a comprehensive water quality, sediment, and biological
monitoring program.
• During the same period of time, the Office of Puget Sound (EPA Re-
gion 10) developed a proposed sound-wide monitoring program which
included goals and objectives, sampling design, operation of the program
and methods for sampling, analysis and reporting of the data (Tetra Tech,
1986a). This document provided a basis for discussions between members
of the Puget Sound Estuary Program (PSEP) technical advisory committee,
scientists, agency staff, and other interested parties.
• The Puget Sound Water Quality Management Plan appointed a Monitoring
Management Committee to define the goals and objectives of the monitor-
ing program and to modify the monitoring strategy proposed by the Office
of Puget Sound. Key aspects of the monitoring program development
included workshops that were held for public and scientific peer review,
including the Technical Advisory Committee (TAG) of the Puget Sound
Estuary Program.
The final report of the Monitoring Management Committee (PSWQA, 1988) summa-
rized the proposed monitoring program and provided explicit statements of the goals
and objectives and expected performance. For each monitoring component the ratio-
nale, methods, replication and statistical sensitivity, and a listing of appropriate
-------
TABLE 2-1. PUGET SOUND AMBIENT MONITORING PROGRAM DESIGN
SPECIFICATIONS FOR SEDIMENT CHEMISTRY SAMPLING (PSWQA, 1988)
Objectives and Rationale
Measuring toxic chemicals of concern will provide data to:
• Assess the potential for sediment toxicity to resident biota.
• Identify areas of Puget Sound that have been, or are, accumulating substantial amounts of toxic chemicals.
• Evaluate temporal changes of toxic chemicals accumulating in sediments.
• Interpret biological and sediment toxicity bioassay data.
Methods
Samples for sediment chemistry will be collected from the upper two centimeters (cm) of sediment, using either a
0.06 m2 box corer or a 0.1 m2 van Veen grab. Three grab samples will be taken at each station and composited. The
same composite will be used for sediment toxicity bioassays and conventional sediment variables. A minimum of the
upper 5 to 10 cm of sediment will be collected for benthic macro-invertebrate abundance determination. Each sam-
pling device has advantages and disadvantages. Although a box corer takes a deeper and possibly less disturbed
sample than does a van Veen grab, the box corer is more difficult and more expensive to use. An evaluation of benthic
sampling equipment for use in PSAMP is in progress.
Variables to be monitored will include selected EPA priority pollutant metals and selected EPA priority pollutant
organic compounds, as well as additional compounds of concern in Puget Sound.
Miscellaneous organic acids and volatile organic compounds will be measured only where a suspected source is
present. Intensive surveys conducted by individual agencies under other programs may be triggered by results from
this program.
Tributyl-tin has recently been implicated as a human health risk (U.S. EPA, 1985). Studies from other parts of the
country have shown accumulations in sediments and animal tissue around large marinas and harbors. The present
concern warrants a comprehensive survey for tributyl-tin in sediments and bottom fish tissue in Puget Sound, but it is
not included in the monitoring program at this time due to inconclusive sampling results from other parts of the
country. Periodic spot checks for this and other contaminants are recommended. Costs of such analyses have not been
included in cost estimates for the ambient monitoring program.
Replication and statistical sensitivity
Replicate samples will not be collected for sediment chemistry, thereby precluding statistical analyses among indi-
vidual stations within a survey. Replicated sampling at all stations was precluded because of the high cost of laboratory
analysis and has not been recommended by Puget Sound Estuary Program sampling and analysis protocols. The
variability of sediment chemistry estimates will be reduced, however, by the compositing technique recommended.
Field and laboratory replication will be required for sediment chemistry samples as part of the quality assurance
program. Stations for field replicates will be chosen so as to be representative of certain areas or embayments and
sediment types.
Statistical analyses may be performed for related groups (clusters) of stations within a survey or for selected stations
over time.
Replicate data from studies on the chemical composition of sediments within the Commencement Bay waterways
indicated that coefficients of variation for several groups of organic chemicals ranged from 17-61 percent (Tetra Tech,
1985). Given a coefficient of variation of 30 percent and three to four replicates (in space or time), the minimum
detectable difference in mean chemical concentration among stations, at the 95 percent confidence level with a power
of 0.8, would be equal to about 100 percent of the overall mean among stations.
Protocols
* Field and Laboratory References: Tetra Tech (1986b, 1986c, 1986d).
* Supporting literature: U.S. EPA (1983), Plumb (1981), U.S. EPA (1982).
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22
sampling protocols were identified. A summary of the design specifications for the
sediment monitoring component of the Puget Sound Ambient Monitoring Program is
presented in Table 2-1. These design specifications provide an example of PSAMP
efforts to clearly state the monitoring objectives and to establish performance criteria.
Four major objectives are stated for the sediment chemistry component of the monitor-
ing effort: assess the potential of sediment toxicity, identify toxic accumulation areas
within Puget Sound, evaluate temporal changes, and provide supplemental data that
will be used to interpret bioaccumulation data and bioassay test results.
The establishment of program requirements, i.e., performance criteria, in terms of the
level of precision and accuracy that is necessary to make decisions regarding the
success of the monitoring effort will define the expectation of the monitoring program.
Issues that must be addressed include: 1) What level of detail will be necessary to
make decisions regarding the success of the CCMP? 2) What level of difference must
be detected in the monitoring program to initiate modifications in the design and
implementation of the monitoring program? This concept of explicitly stating perfor-
mance criteria is the cornerstone of the systems design approach that is described
below. The explicit statement of the monitoring program requirements provides a
basis for evaluating expected and actual monitoring program performance (Section
5.0). Evaluation of the effectiveness of alternative monitoring designs also provides a
basis for discussion of alternative monitoring approaches and sampling layouts.
Performance criteria are addressed in the design specifications for the sediment
chemistry sampling of the Puget Sound Ambient Monitoring Program (Table 2-1).
Under the discussion of replication and statistical sensitivity, basic information is
provided on background variability and the minimum detectable difference between
groups of stations. However, the specification of performance criteria associated with
monitoring program designs should be made more explicit than those presented in
these design specifications. For example, one of the stated objectives is to evaluate
temporal changes in toxic accumulation. Corresponding performance criteria should
be developed by determining the magnitude of trend that can be expected to be de-
tected with the planned level of sampling effort and the period of time that will be
required to detect statistically significant trends. The use of historical data and the
application of statistical methods to evaluate and establish decision criteria are ad-
dressed in Section 5. An example of the use of statistical methods to determine the
magnitude of trends that can be measured in a particular monitoring design is pre-
sented in Case Study 2 (Appendix A2.0).
2.2
Performance
Criteria
_
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23
2.3 The Quality Assurance Management Staff (QAMS) of EPA has developed an ap-
Additional proach to designing data collection programs based on the development of Data
Guidance Quality Objectives (DQOs). This approach has many of the same elements as the
systems approach for designing monitoring programs that are discussed in this docu-
ment. The DQO process places emphasis on defining the objectives of data collection
programs, specification of the decisions that will be made with the data collected, and
the possible consequences of the decisions being incorrect (QAMS, 1986). A key
aspect to the development of DQOs is the evaluation of the desired degree of certainty
in conclusions to be derived from the data. This evaluation is similar to the process
described in this document for the development of decision criteria. In both methods,
the decision criteria are developed to ensure that adequate information is obtained for
making decisions. A case study describing the use of DQOs in Superfund Remedial
Investigations is presented by Neptune et al. (1990).
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24
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25
3.0 Establish Testable Hypotheses and
Select Statistical Methods
Table 3-1 provides examples of monitoring objectives that are common to many NEP
monitoring programs. The range of corresponding questions provides some idea of the
breadth of monitoring issues that are encompassed in the monitoring objectives.
In designing the monitoring program, these broad objectives must be focused to
identify specific monitoring variables and corresponding monitoring activities.
ESTABLISH TESTABLE
HYPOTHESES AND SELECT
3.1 The questions identified in Table 3-1 give rise to a number of alternative scientific
Establish Testable hypotheses. For example, questions regarding the water quality component of a
Hypotheses monitoring program and hypoxia can lead to several hypotheses regarding the response
of dissolved oxygen concentrations in the water column. Example hypotheses include:
• Nutrient reduction strategies will result in increased dissolved oxygen
concentrations throughout the estuary.
• Any increases in dissolved oxygen concentrations will be long term,
improvements will not occur for 20 - 30 years.
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26
It is clear that the monitoring program must be designed to address a wide range of
alternative hypotheses. The recommended procedure for ensuring that sufficient
information and the right type of information is developed in the monitoring program
is to specify, prior to the collection of any samples, the statistical model that will be
used to analyze the resulting monitoring data, and to specify testable (null) hypoth-
eses.
TABLE 3-1.
EXAMPLES OF MONITORING PROGRAM OBJECTIVES
AND ASSOCIATED QUESTIONS
Objective
Document response of water quality
variables to management actions
Characterize spatial and temporal
patterns in bioaccumulation
Monitor the status of the ecosystem
Questions
Are nutrient reduction strategies effective?
Is the risk of hypoxia reduced?
Is there a decrease in phytoplankton
biomass?
Is light transmittance affected?
Is fish community structure affected?
What is the risk of consuming seafood
products from within die estuary"?
Are there trends in fish and sheUfish
contaminant concentrations?
Do toxic hot spots exist and what are
the influences on bioaccumulation?
What is the relationship between
sediment concentrations of contaminants
and observed tissue concentrations?
What is the relative contribution of
different sources of pollutants?
What are the trends in selected indicators?
What are the consequences of the
physiological, morphological and
molecular changes on which indicators
are based on organism survival and
population health?
What is the status of species of
commercial and recreational importance?
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27
For example, expanding on the water quality issues above, the process of selecting the
statistical model and testable hypotheses for detecting trends in dissolved oxygen
concentration is outlined below.
A regression model is used to partition field observations (Xy) between several factors
that potentially influence dissolved oxygen concentrations:
where:
Xy = field observation from time i and location j of dissolved oxygen
concentration at a specified depth
Po = mean of all Xy observations
= temporal component of the measurement
(32lj = spatial component of the measurement
P3tilj = location-time interaction component of the measurement
£y = random errors not accounted for by p0, pjtj, p2lj, p3tjl:
One possible null hypothesis to be tested in this case is that there is no temporal trend
in measured dissolved oxygen concentrations [H0: Pj = 0]. If it is concluded that H0 is
false, then an alternative hypothesis [Ha: p1 is positive (increasing trend) or negative
(decreasing trend)] will be assumed to be true. The specification of this model also
provides the basis for addressing other scientific hypotheses of interest. For example,
is there evidence of location-time interaction [H0: p3 = 0]?
Green (1979) emphasizes the importance of developing testable hypotheses during the
design phase of environmental studies and points out that hypothesis formulation is a
prerequisite to the application of statistical tests. As pointed out below in Section 5,
the development of testable hypotheses and the selection of statistical methods are also
the first steps in evaluating the expected performance of the monitoring program. For
the dissolved oxygen example, this means determining the minimum trend that can be
detected for a specified level of sampling effort. The process of devising testable
hypotheses should also be used to initiate discussions between the management
committee and technical experts on the importance of individual monitoring objectives
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28
and the relevance of the associated questions. Platt (1964) provides an excellent
review of the importance of hypothesis development in the design of scientific investi-
gations in general.
The statistical tests that will be used to analyze the resulting data must be specified for
each hypothesis developed. The applicability of univariate, multivariate, parametric
and nonparametric methods must be carefully evaluated. Selected references that
provide background information on the use of various statistical methods are summa-
rized in the annotated list presented in Table 3-2. These references will provide
program coordinators and scientific staff with a basic understanding of the analytical
options available. However, it is essential to involve the statisticians responsible for
the analysis of the data in both the development of testable hypotheses and the selec-
tion of analytical methods.
The statistical software that will be used to analyze the data should also be identified at
this step in the design process. Berk (1987) describes the attributes of effective
microcomputer statistical software. Meads (1990) summarizes the results of a user's
survey of six advanced statistical packages available for PCs. The American Statisti-
cian, a publication of the American Statistical Association, also regularly reviews new
statistical software and updates to existing packages.
3.2
Selection of
Statistical
Methods
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29
*' '"T5. ^
^v?' '•^"" "4».'-- .•••••••••i ^
Sfjf «< s s ^-^Xx f f * s ^Vis-iv *•
ii\s^™ *C.W^«™.
TABLE 3-2,
REFERENCES TO BASIC MONITORING DESIGN AND STATISTICAL TEXTS
Reference
Monitoring Design
* Sampling Desip and Statistical Methods for
Environmental Biologists (Green, 1979)
Statistical Methods for Environmental
Pollutiori Monitoring (Gilbert, 1987)
Sampling Techniques (Coehran, 1977)
» Statistical I*rineiples in Experimental
Design (Winer, 1971)
General Statistics
* Biometry (Sokal and Rohlf, 1981)
» BiostatisficalAnalysis'CZar, 1974)
• Applied Statistics, Principals and Examples
(Cox and Snell, 1981)
Multivariate Statistics
* Applied Multivariate Statistical Analyses
(Johnson and Wichem, X982)
' Applied Regression Analysis
(Draper and Smith, 1981)
« Multivariate Statistical Methods: A Primer
(Manly, 1986)
Description
• Introduction to principles and options for sampling and
statistical design. Examples of monitoring design and
application of statistical methods.
• General reference that describes a wide range of statistical
, methods and their application. Intended for nonstatisticians.
Description of several statistical techniques that are not
- commonly seen in general references.
r S
• Comprehensive review of sampling methods and theory.
Detailed presentation of topics at aa introductory level.
Applicability not limited to environmental sampling and
monitoring.
• Description of hypothesis testing concepts.
Basic reference to statistical techniques most frequently used
in the biological sciences. Numerous examples of applications.
Basic reference to statistical techniques most frequently ased
in the biological sciences. Emphasis on analysis of variance
techniques.
In depth examples of many of the most common
statistical applications.
General introduction to multivariate methods.
Detailed introduction to regression techniques. Includes
section on planning large regression studies.
Brief description of the most common multivariate
techniques, with examples*
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31
""•V
4.0 Select Analytical Methods and
Alternative Sampling Designs
The goal of this step in the design process is to develop detailed monitoring program
specifications. In addition to the statistical methods, these specifications include the
field collection and laboratory analysis methods for individual monitoring variables,
and the appropriate quality assurance/quality control procedures. Alternative sampling
layouts including numbers and location of sampling points, sample frequency, and the
level of sample replication should also be developed. This information will then be
used in the next step (see Section 5) to evaluate expected monitoring program perfor-
mance and to select the most efficient sampling layout among the alternatives.
SELECT ANALV
H08$Af«rAlHl
SAMPLING OESJQNS
4.1
Selection of Field
and Laboratory
Methods
Appendix B of this document provides descriptions of numerous sampling methods
that are routinely utilized in estuarine monitoring programs. These descriptions
include information on how the data can be used to address the goals of the monitoring
programs and to evaluate the success of the CCMP. The essential elements of quality
assurance and quality control programs are also described. The purpose of selecting
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32
field and laboratory methods at this stage of the design process is to ensure: the
feasibility of using the selected methods in conjunction with the proposed level of
sampling effort; that any data used to evaluate expected monitoring performance, a
crucial step in the design process, are directly comparable with data that would be
collected in the proposed monitoring effort; and, that standardized methods are used.
The development and application of an effective QA/QC program is discussed
throughout the appendix. The Chesapeake Bay Split-Sample Program (U.S. EPA,
1991) is presented as a valuable approach to maintaining QA/QC.
Standardized protocols or performance criteria should be developed to ensure that the
data collected by the different groups participating in the estuary monitoring program
arc directly comparable. Becker and Armstrong (1988) describe the process of devel-
oping standardized sampling and analysis protocols for measuring selected environ-
mental variables in Puget Sound. The key feature of the development of these proto-
cols was a series of workshops at which regional scientists and managers worked
together to develop standardized methodologies. The protocols that have been devel-
oped are described in the introduction to Appendix B. These protocols were also used
as a primary source of information for the description of methods that are included in
Appendix B.
In the development of alternative sampling layouts, consideration should be given to 4.2
the trade-offs between the benefits of a comprehensive monitoring effort and available Alternative
funding. In general, federal support for these programs will be limited both in the Sampling Designs
amount of funding available annually and the duration of funding. Consideration
should be given to both limiting the scope of monitoring efforts and making the most
efficient use of ongoing monitoring programs in the estuary.
Ideally, the scope of the program should be adequate to meet all identified monitoring
objectives. Where the funding is not available to meet all the objectives, however, the
individual monitoring components should be prioritized on the basis of the relative
importance of related management issues and the availability of existing information.
Generally, emphasis should be placed on focusing monitoring efforts in order to attain
the level of precision necessary to evaluate the effectiveness of individual management
actions, rather than implementing a comprehensive monitoring program that lacks the
ability to detect the level of changes expected over time.
Given the wide variety of habitats in individual estuaries, the large variability gener-
ally associated with environmental samples, and the limit of funding, alternative
sampling strategies should be investigated. Through design optimization, the sampling
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33
effort can be distributed spatially and temporally in such a way as to maximize the
amount of information obtained within the area sampled. The strategy behind most
sample design optimizations is either to minimize the detectable difference or trend for
a fixed cost or to minimize the cost for a specified minimum detectable difference or
trend. The strategy adopted will depend upon the specific situation for each monitor-
ing program. In either case, the goal is to obtain the maximum amount of information
per dollar spent.
The choice of a sampling design depends on several factors including the objectives of
the monitoring program, the type of data that are required to test the null hypothesis,
the underlying assumptions of statistical tests, and the spatial and temporal distribution
of the monitoring parameters. These factors can affect both the validity of the test
results and the efficiency of the monitoring program (cost to obtain a given level of
detection). A brief summary of common sampling designs is presented in Figure 4-1.
Figure 4-1.
Description of various
sampling designs.
Sampling Designs
Simple Random: Samples are independently located
at random
Systematic: Samples are located at regular
'•• intervals
Stratified: The study area is divided into
nonoverlapping strata and samples
are obtained from each
Multistage: Large primary units are selected,
which are then subsampled
' • •
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84 •
MMS"1"1 ^ 1
*|l»l 111*111'l :
Sw(l*«*wj4rtj*wwwMm4^w*
"•X.
The most basic method of collecting monitoring data is simple random sampling. With
this design, samples are selected randomly and with equal probability. While this
method is easy to implement, there are a variety of sampling designs that can be more
efficient. Such designs will produce estimates with smaller standard errors for the
same sampling effort, or require fewer samples to obtain the same standard error as
would be obtained with simple random sampling.
One such design is systematic sampling. In systematic sampling, sample units are
selected at fixed intervals in space or time, usually with a random start. There are
many variations of systematic sampling; however, they all share some common
advantages and disadvantages. The even coverage obtained with systematic sampling
tends to ensure that each sample, on average, is more representative of the population
as a whole than a simple random sample. Therefore, such samples tend to have a
smaller standard error associated with them. Problems can arise that lead to bias or
increased variance if there is an underlying pattern or periodicity in the population
over space or time, which is common with environmental data. In addition, it can be
difficult to obtain a valid estimate of the standard error if the data cannot be assumed
to be distributed randomly.
Another design often used is stratified sampling. By dividing the study area into
nonoverlapping homogeneous strata it is possible to optimize the sampling effort in
several ways. First, the samples can be allocated to the different strata in proportion to
the size of the strata and the variability within the strata, and in inverse proportion to
the cost of sampling in those strata. This method will insure that the minimum vari-
ance will be obtained for a given cost. Other criteria, such as the ecologic importance
of strata and the parameters being measured, can also be taken into account when
allocating sampling effort. Stratified sampling also allows the use of the best sampling
designs within strata to further increase the sampling efficiency. Stratified sampling
works well in a tiered approach because it allows monitoring performance assessment
and design modification to be made on a stratum by stratum basis. Stratified sampling
also yields estimates for each stratum, providing information that better represents the
area being sampled, and is therefore more ecologically meaningful. As an example,
two strata may each have a significant trend for a given parameter, but the trends may
be in opposite directions. If the data were combined (such as in systematic sampling),
the trends might cancel each other out and result in a conclusion of no significant
trend. Because stratified sampling ensures that some samples will be taken from each
Stratum, over the entire study area, it helps ensure that the overall estimate will be
more representative on average than one obtained from a simple random sample. This
advantage of stratified sampling will be realized even if optimal allocation is not used
and the strata are defined arbitrarily with respect to the parameters of interest. In
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35
1 SSSSHS
general, if the variability within individual strata is less than the overall variability in
all combined strata, the standard errors obtained with stratified sampling will be less
than those obtained from systematic sampling, which will be less than those obtained
from simple random sampling. Applications of stratified sampling to environmental
studies are found in U.S. EPA (1982a), Jensen (1973) and Reckhow and Chapra
(1983).
Multistage sampling is another cost effective method of allocating sampling effort over
large areas. In this design, large primary sampling units are composed of smaller
secondary units. For large-scale studies, third stage units may also be used. Within
each of the selected first stage units, one or more second stage units are selected. In
addition to increased sampling efficiency, this method allows intensive sampling to be
done only at the second stage, while parameters that are inexpensive to measure can be
obtained for the first stage units. This provides some level of monitoring over a wide
area, and if problem areas are detected, the distribution of second stage units can be
reallocated. The information collected from the first stage units can also be used to
implement variable probability sampling at the second stage to further increase the
sampling efficiency.
Further information on these sampling designs and methods for calculating required
sample sizes and optimal distribution of samples can be found in Gilbert (1987) and
Cochran (1977).
4.3
Use of Existing
Monitoring
Programs
A monitoring strategy that incorporates ongoing monitoring programs or elements
from these programs can significantly reduce the cost of the monitoring effort. Exist-
ing compliance and resource monitoring programs may produce data that can com-
pletely satisfy, or augment, the spatial and temporal coverage required by an NEP
monitoring program. Additionally, by adopting sampling and analytical methods of
ongoing monitoring programs as standard protocols for NEP monitoring program
components, data from these existing monitoring programs may be used in evaluating
the effectiveness of the CCMP.
An inventory and evaluation of existing federal, state, local, and volunteer monitoring
programs within the estuary drainage basin should be conducted in order to assess their
usefulness and applicability in evaluating the effectiveness of the CCMP. Key tasks of
this assessment process include:
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36
Identification of existing and planned programs as well as special projects
that may contribute data useful in evaluating the effectiveness of the
CCMP
• Determination of whether NEP monitoring program objectives could be
cost-effectively met by incorporating sampling and analytical methodolo-
gies from these existing monitoring programs
Specific monitoring variables, statistical and analytical methods, and quality assur-
ance/quality control (QA/QC) protocols selected will depend upon the stated objec-
tives and action plans outlined in the CCMP, and specific hypotheses to be tested in
the monitoring program. Therefore, techniques from existing monitoring programs
may not always be adopted. However, whenever possible, it is recommended that
programs work to ensure comparability of their methods with existing programs.
Several specific opportunities for taking advantage of ongoing monitoring efforts are
described below.
NOAA's National Status and Trends (NS&T) Program
NOAA's National Status and Trends Program is designed to assess the current condi-
tions of environmental quality in the nation's coastal zone and to determine whether
these conditions are improving or deteriorating (NOAA, 1989). In order to achieve
this objective, NOAA currently supports the Benthic Surveillance Project and the
Mussel Watch Program.
The Benthic Surveillance Project regularly measures concentrations of contaminants in
sediments and tissues of bottom-dwelling fish. The occurrence of external and internal
symptoms of disease (e.g., fin erosion and liver tumors) are also documented. The
Benthic Surveillance Project currently collects and analyzes sediment and bottomfish
samples at 75 estuary sites.
The Mussel Watch Program collects mussels and/or oysters once a year from approxi-
mately 200 sites nationwide (NOAA, 1989). Analyses of sediment and bivalve tissue
concentrations of trace metals, DDE, PCBs, aromatic hydrocarbons, and radionuclides
arc conducted.
NS&T has developed standardized methods for analysis, quality assurance (QA), and
quality control (QC) for fish and shellfish tissue and sediment contaminants. The
methods employed in this program and sampling locations should be reviewed to
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I37
S WKW^
detennine the feasibility of their incorporation into the planned monitoring efforts in
each estuary. For detailed information concerning analytical and QA/QC methods,
contact:
NOAA National Status and Trends Program
NOAA N/OMA32
6001 Executive Blvd
Rockville,MD 20852
In addition, NOAA is establishing a National Estuarine Reserve Research System
(NERRS). National Estuarine Research Reserves are established and managed for
long-term environmental monitoring and scientific research. The results of scientific
research at the Reserves are important sources of information and data to support
coastal zone management and decision making. For more information concerning the
NERRS, contact:
Marine and Estuarine Management Division
Office of Ocean and Coastal Reserve Management
NOS/NOAA
1825 Connecticut Avenue, NW
Washington, D.C. 20235
EPA's Environmental Monitoring and Assessment (EMAP) Program
The goal of EMAP's Near Coastal (EMAP-NC) Program is to monitor the condition of
near coastal ecosystems, evaluate the relationship between ecological condition and
anthropogenic disturbance, and assess the effectiveness of pollution control actions
and environmental policies on a regional and national scale. EMAP will provide
information on biological and chemical indicators, sampling design and methods,
analytical methods, and QA/QC protocols.
EMAP-NC sampling design consists of three schemes (U.S. EPA, 1990b). EMAP's
regionalization scheme divides the nation's estuaries and coastal resources into bio-
geographical provinces (e.g., Virginian, Carolinian, Louisianian, Acadian,
Columbian, and Californian provinces). EMAP's classification scheme classifies
estuaries into three resource classes that have similar physical features:
• Large, continuously distributed estuaries (e.g., Chesapeake Bay, Long
Island Sound)
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38
• Large, continuously distributed tidal rivers (e.g., Potomac, Delaware,
Hudson Rivers)
• Small, discretely distributed estuaries, bays, inlets, and tidal creeks and
rivers (e.g., Barnegat Bay, NJ; Indian River, FL; Lynnhaven Bay, VA)
EMAP's sampling scheme consists of elements of systematic, random, and fixed
location sampling. Large, continuously distributed estuaries are sampled using a
randomly placed systematic grid, with grid points about 18 km apart. Large tidal
rivers are sampled along systematically spaced lateral transects. Transects are located
about 25 km apart. Two sampling points are located on each transect, one randomly
selected, and one selected using scientific judgement to identify sampling locations
that may be indicative of degraded conditions in the system. Small estuaries are
sampled by partitioning them in groups of four, selecting one estuary randomly from
each group of four, and sampling at two stations in each small estuary selected.
EMAP will operate on a four year sampling cycle, with one-fourth of the sites in a
region sampled each year. Sampling will be undertaken only during the months of
July and August. Given this sampling design and schedule, it is clear that a very small
amount of EMAP data, if any, is likely to be available to most NEP estuary programs
in a particular year. The stratified random sampling approach developed by the EMAP
program may be of use to some NEP estuary programs if applied on a much smaller
scale. The EMAP program is currently evaluating the advantages of sampling on
alternative spatial scales.
EMAP-NC indicators of environmental quality are described in EMAP's Near Coastal
Program Plan for 1990 and Ecological Indicators documents (U.S. EPA, 1990b and
1990c). Ecological indicators include:
• Response indicators
- Benthic species composition and biomass
- Gross pathology of fish
- Fish community composition
- Relative abundance of large burrowing shellfish
- Histopathologyoffish
- Apparent redox potential discontinuity
* Exposure indicators
- Sediment contaminant concentration
- Sediment toxicity
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39
- Contaminants in fish flesh
- Contaminants in large bivalves
- Water column toxicity
- Continuous and point measurements of dissolved oxygen concen-
tration
• Habitat indicators
- Salinity
- Sediment characteristics
- Water depth
• Stressor indicators
- Fresh water discharge
- Climate fluctuations
- Pollutant loadings by major category
- Land use patterns of watershed by major category
- Human population density/demographics
- Fishery landings statistics
Methods for collecting many of these indicators are addressed in the methods section
(Appendix B).
The information from EMAP-NC is intended to establish ecological trends over the
time frame of decades. Although this information may not be immediately available to
the National Estuary Program, EMAP-NC does provide valuable information and
assistance in the areas of sampling design, indicators, sampling methods, quality
assurance and information management. Additionally, EMAP-NC is implementing a
secondary monitoring strategy that focuses on local and state issues. The objective is
to foster the incorporation of EMAP-compatible monitoring efforts for near-coastal
resources into sampling by state and local agencies. Such efforts are intended to build
upon the present EMAP design to permit assessment of environmental conditions at
spatial and temporal scales not presently addressed by EMAP.
For detailed information concerning EMAP analytical, statistical, and QA/QC meth-
ods, consult the Director of EMAP's Near Coastal Waters Monitoring Program at:
Environmental Research Laboratory
U.S. Environmental Protection Agency
27 Tarzwell Drive
Narragansett, RI02882
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40
Local Monitoring Programs
In addition to ongoing compliance and resource (e.g., fisheries) monitoring that is
conducted in each estuary, there are also other opportunities to augment the estuary
monitoring programs. For example, states are required under 305(b) of the Clean
Water Act to conduct water quality assessments, and they are encouraged to include
assessments of trends in their reports that are submitted biennially to EPA (U.S. EPA,
1989b). Other monitoring programs are conducted by state agencies, universities and
pollutant dischargers. These programs should be evaluated to determine their use in
monitoring CCMP implementation.
State sponsored and private volunteer monitoring programs have been shown to be
very valuable in collecting data on estuarine water quality, beach litter, marine mam-
mal strandings, and the status of other estuarine resources (Armitage et al., 1989).
Volunteers in the Chesapeake Bay watershed have collected data used to help verify
water quality models and to identify correlations between measured variables, such as
low dissolved oxygen, and the frequency of observed events such as fish kills and
algae blooms (Ellett, 1988).
Volunteer monitoring programs have also been useful in developing an educated and
involved constituency committed to protecting water resources (U.S. EPA, 1990d).
Partnerships between the public and government agencies responsible for management
must be developed if National Estuary Program CCMPs are to be effectively imple-
mented. Establishment of volunteer monitoring programs has proven to be an effec-
tive way to build public commitment to achieving environmental quality goals and
objectives in the CCMP. Through participation in monitoring programs, citizens learn
how they contribute to pollution problems and develop a sense of stewardship toward
the waters they are monitoring. Volunteer monitoring programs also help the public to
understand the difficulties faced by the scientific community in linking water quality
changes to impacts on living resources.
Each National Estuary Program Management Conference should establish a volunteer
monitoring component as an integral part of its monitoring program. The experience
of citizen monitoring programs throughout the country proves that volunteers can be
trained to carry out a wide variety of environmental monitoring tasks, provided they
are given the appropriate equipment and instruction. In estuaries, volunteers can be
especially helpful in upstream areas not normally covered by a state's monitoring
network. Basic water quality measurements such as pH, transparency, salinity,
dissolved oxygen, and temperature can provide useful information to the comprehen-
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41
sive monitoring program. Trained volunteers can also be used to assess aquatic
vegetation in the estuary and can provide information on acute problems such as spills,
fish kills, and algae blooms. EPA has published a guidance document describing how
to plan and manage effective volunteer environmental monitoring programs (U.S.
EPA, 1990d). The document provides an overview of the use of citizen volunteers in
environmental monitoring. It discusses how to plan and organize volunteer monitoring
projects, how to involve the media, and how to prepare quality assurance plans for
volunteer programs.
In general, estuary monitoring programs must select cost-effective methods that
provide data essential to assessing the effectiveness of the CCMP. Whenever possible,
programs should work to ensure comparability of their methods with applicable
ongoing programs. Key aspects of the incorporation of existing state agency and
volunteer monitoring programs into the overall design for the Puget Sound Ambient
Monitoring Program are described in Case Study 1 (Appendix Al.O).
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43
•.*
*««f<[J
JV AlAHV&fS A
5.0 Evaluate Monitoring Program
Performance
Although often overlooked and neglected, the evaluation of monitoring program
performance is potentially the most important step in the design and review process.
The performance evaluation motivates the development of explicit statements of
program objectives as well as the specification of quantitative performance criteria
during the design phase. During the course of the monitoring effort, performance
evaluations provide a systematic procedure for measuring success in terms of the
ability to meet program goals. The periodic evaluation process also identifies the need
to modify sampling design and methods.
Evaluation procedures are essential because the information developed in the monitor-
ing programs must be sufficiently precise and scientifically defensible. The monitor-
ing programs will provide the primary source of information that will be used to
evaluate the success of the CCMP. This information will be used as a basis for
determining the efficacy of selected management strategies and the accuracy of model
predictions upon which many management decisions have been based. The monitor-
ing programs will also provide quantitative information that will guide decisions
regarding needed modifications to the management plan.
Additionally, the cost of the estuary monitoring programs will be substantial. In order
to protect this investment, it is essential to assess expected performance prior to
collecting the first samples. This performance information will provide the basis for
determining the feasibility of proposed sampling strategies, selecting the most effec-
tive monitoring components and variables, and optimizing the overall monitoring
effort.
The two types of performance evaluations are shown in Figure 1-1 and highlighted in
the schematic on the next page. The first is the evaluation of the expected perfor-
mance of individual components of the monitoring program (e.g., the evaluation of
trends in toxic chemical accumulation in sediments). The first evaluation takes place
during the design phase. The second type of performance evaluation is the assessment
of overall program performance. This assessment takes place after the monitoring
program has been implemented (e.g., after the first full year of data have been col-
lected). The objective is to determine if the overall goals of the program are being met
by the individual monitoring components and if the program should be modified by
adding, deleting or expanding the scope of individual monitoring components. The
essential feature of both types of evaluation is the existence of a feedback loop that
provides the pathway for modifying the system's design based on monitoring program
performance.
-------
The establishment of performance criteria (e.g., the ability to detect a change in
chlorophyll concentrations of 5 pg# over a period of five years) is a fundamental part
of developing monitoring objectives. As indicated in Section 2.0, these performance
criteria represent the level of change that must be detected in order to make manage-
ment decisions regarding the effectiveness of the CCMP. It is these performance
criteria that will be used to evaluate the applicability of individual components of the
monitoring program. The specification of sampling methods for proposed monitoring
program components described in Step 3 (Section 4.0) includes the development of
alternative sampling strategies, including the monitoring variables/indicators and the
level of sampling effort (numbers of sampling stations and sample replicates). The
goal of the performance assessment is to evaluate the effectiveness of these alternative
sampling designs in terms of the established performance criteria. The results will
5.1
Evaluate the
Expected
Performance of
Individual
Monitoring
Program
Components
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45
j. * 0
. *\ »»\ , '•=•<"•• *-
f vyAWiW
provide the basis for determining the relative benefits of individual monitoring compo-
nents and selecting the final monitoring design.
The questions that will be addressed in these analyses are:
• Can the proposed sampling effort meet the needs of the monitoring pro-
gram as defined in the performance criteria associated with the stated
objectives?
• How can the proposed program be modified to ensure that these objectives
are met?
As indicated by the feedback loop shown in Figure 1-1, this is an iterative process.
Proposed sampling designs are evaluated and modified, if necessary, to meet the
overall objectives. The tools for conducting these analyses are described in Sec-
tion 5.3.
5.2
Evaluate Overall
Program
Performance
The overall performance of the monitoring program should be evaluated at periodic
intervals. Initially, this evaluation should take place at the conclusion of the first year
of sampling. This evaluation should compare the results with the expected monitoring
performance, and a list of required modifications should be prepared. Opportunities
for streamlining the program should be identified, and the performance criteria should
be reviewed and revised, if necessary, for subsequent evaluations.
5.3 The primary tool for conducting these analyses is statistical power analysis. Statistical
Statistical Power
Analysis Methods
power analysis provides an evaluation of the ability to detect statistically significant
differences in a measured monitoring variable. The importance of these analyses can
be seen in the examination of the possible outcomes associated with testing the null
hypothesis (e.g., H0: sampling location has no effect on observed sediment contami-
nant concentrations) shown in Figure 5-1:
• The null hypothesis is true, and it is rejected. This is referred to as a Type I
error (a) and is commonly called the significance level of the test By
convention this value is routinely set at 0.05, i.e., the investigator accepts a
small probability of incorrectly concluding that there are differences in
sediment contaminant concentrations between sampling locations. Lower or
higher values of the significance level of the test may be appropriate depend-
ing on the consequences of incorrectly rejecting a true null hypothesis.
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46
'""5.
g ACCEPT
GO
O
Q REJECT
HYPOTHESIS
ACTUALLY TRUE ACTUALLY FALSE
1-CC
/^^Jf ;.^
^ •>, \xS."
.. % , v.} ' ' i f ^
..,>...•:.•:•:..•:... .... ... . .
1-P
Figure 5-1. Hypothesis
testing: possible circum-
stances and test outcomes.
• The hypothesis is true, and it is accepted. This is the complement of the
Type I error (I- a).
• The hypothesis is false, and it is accepted. This is referred to as a Type II
error (p). While the significance level of the test is consistently reported
with the results of statistical tests, the probability of accepting the null
hypothesis when it is not true (Type II error) is almost never reported with
statistical test results. Moreover, the consequences of the Type II error are
probably not fully comprehended by many investigators.
• The hypothesis is false, and it is rejected. This is the complement of the
Type II error (1- p) and is referred to as the power of the test. Therefore,
statistical power is the probability of correctly detecting an effect.
Consideration of these possible outcomes of a statistical test leads to three important
conclusions. The first is that failure to reject the null hypothesis does not justify its
acceptance. For example, the failure to reject the null hypothesis (e.g., that location
has no effect on sediment contaminant concentrations) may occur either because there
really is no effect (probability = 1 - a) or because the power of the test is so low
(probability = p). In the later case, a Type II error occurs because the statistical test is
weak. This may be due to the highly variable nature of the sampling environment or
the low level of sampling effort. In either case, it is possible to evaluate the probabil-
ity of the Type II error for any statistical comparison of environmental data. There-
fore, the second conclusion is that the probability of the Type II error should be
reported with all statistical test results. The third conclusion is most relevant to the
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K-
r
47
design of the NEP monitoring programs - the expected power of the statistical test
should be evaluated prior to implementing the sampling program.
Although statistical power analyses are not routinely conducted with statistical tests of
significance, there is ample guidance available and the tests can be easily performed.
Power analyses can be conducted using standard equations and tables or nomographs.
The basic tools for conducting these analyses are provided by Cohen (1977), Dixon
and Massey (1969), and Winer (1971). More comprehensive tables and nomographs
are provided by Pearson and Hartley (1951), Tang (1938), Lehmer (1944), and Scheffe
(1959), but the use of these references requires more advanced statistical training.
There are also a wide range of computer programs available for conducting statistical
power analyses. Goldstein (1989) reviewed several MS/PC-DOS power analysis
programs and compared the methods they cover, their ease of use, graphics capabilities
and their computational accuracy. Additionally, a statistical power analysis tool for
analysis of variance is available on the EPA's Ocean Data Evaluation System (U.S.
EPA, 1987a), and the U.S. EPA guidance for conducting fish histopathology studies
(U.S. EPA, 1987b) provides nomographs for the power analyses associated with
contingency table analysis.
Most basic statistical texts address hypothesis testing and the concepts of statistical
power analysis. The statistical texts by Dixon and Massey (1969) and Winer (1971)
provide introductory level descriptions as well as the tables for conducting these
analyses. The statistical text by Scheffe (1959) provides a theoretical description of
power tests for the analysis of variance.. The Technical Support Document for the
ODES Statistical Power Analysis Tool (U.S. EPA, 1987a) provides a detailed intro-
ductory level description of power tests for the analysis of variance.
The power of all statistical tests is dependent upon the following design parameters:
• significance level of the- test (a)
• level of sampling effort (i.e., number of sampling stations and sample
replicates)
• minimum detectable difference in the effect that can be detected
• natural variability within the sampling environment
This relationship between the power of a statistical test and the design parameters
makes several types of power analyses possible. The power of the test can be deter-
mined as a function of any of these design parameters. Alternatively, the value of any
individual design parameter required to obtain a specified power of a statistical test can
be determined as a function of the other parameters.
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48
The results of statistical power analyses are generally reported in two formats. In the
first, shown in Figure 5-2, the minimum difference in the effect that can be detected is
shown as a function of level of sampling effort (number of replicate samples). The
results of this type of analysis provide a quantitative comparison of alternative sampling
layouts, and they are especially useful in the evaluation of proposed NEP monitoring
programs. Using these results, the level of sampling effort required to obtain a selected
level of precision in the monitoring program can be determined. This example was
taken from the evaluation of bioaccumulation monitoring strategies (U.S. EPA, 1987c).
Historical data for liver concentrations of PCBs in winter flounder were used to evalu-
LLJ
in
o
UD
cc
01
Q
LU
CO
s
LU
tD
a
2
2
350 n
300-
250-
200-
150-
100-
50-
Fixed Design Parameters
Statistical Significance (a) = 0.05
Power (1-p) =0.80
Stations = 4
Estimated Variance (a2) = 2.06
4 6 8 10 12
NUMBER OF REPLICATES
14
16
Figure 5-2. Minimum
detectable difference vs.
number of replicates for
fixed set of design
parameters.
-------
« v. ""?•*• 1 "s •,
.' .-'.,
49
ate the expected performance of alternative sampling designs. The results shown in
Figure 5-2 were taken from the first phase of the analyses. They indicate that the
minimum difference in tissue concentrations of PCBs that could be detected between
sampling locations with the collection and analysis of tissue from five fish at each
location was approximately 120 percent of the overall mean at all stations, or 4.9 mg/
kg. Additionally, these results also show that the sampling program would require the
collection and analysis of tissue from seven fish at each location to detect a difference
equal to the overall mean among sampling locations. Based on the results of these early
analyses during the design phase of the monitoring program, alternative sampling
strategies were evaluated. Additional power analyses were conducted to evaluate the
compositing of fish tissue prior to laboratory analyses. The results indicated that the
collection of replicate composite samples would permit the detection of substantially
smaller differences in tissue concentrations of PCBs at a much lower cost.
Statistical power analyses may also be displayed as shown in Figure 5-3. The power
of the test (the probability of detection) is shown as a function of the minimum detect-
able difference that can be detected between locations or over time. These results,
taken from Case Study 2 (Appendix A2.0), were obtained from the analysis of a subset
of water quality data that were collected in Chesapeake Bay. As part of the 1987
Figure 5-3.
Power vs. minimum
detectable difference.
0.0
0.00 0.04 0.08 0.12 0.16 0.20
Minimum Detectable Difference (SLOPE, mg/4-yr)
-------
50
-v.\
«/',.
Chesapeake Bay Agreement, a commitment was made to reduce the total inputs of
nitrogen and phosphorus entering the mainstream of the Chesapeake Bay by at least
40% by the year 2000. The target of 40% nutrient reduction was developed by model-
ing the environmental conditions in the Chesapeake Bay with a two-dimensional,
steady-state model. The modeling results indicated that, after full implementation of
the 40% reduction goal, the lowest average dissolved oxygen would be 1.6 mg/t and
no waters would be anoxic. Therefore, the minimum performance criterion for the
monitoring program should be the ability to detect a difference in dissolved oxygen
equal to 1.6 m/l.
To test the ability of the existing monitoring program to meet the performance crite-
rion, historical data were used to estimate measurement variability. Statistical power
analyses were then conducted, using estimates of the maximum and minimum vari-
ance. The results shown in Figure 5-3 indicate that the minimum trend in dissolved
oxygen concentration that can be detected with a probability of 0.80 and ten years of
data is on the order of 0.06 to 0.13 mg/l-yr. These results indicate that the existing
monitoring program would meet the specified performance criterion.
The relationship between power and minimum detectable difference, shown in Fig-
ure 5-3, provides the information required to evaluate the probability of a Type II error
and the probability of detecting specific levels of effects in a proposed sampling
program. Recent examples of the application of this type of power analysis include:
Parkhurst (1985); Toft and Shea (1983); and Rotenberry and Wiens (1985). The
necessity for this type of analysis in the evaluation of statistical test results is provided
byPeterman(1989).
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51
iwXw,v w
6.0 Design and Implement Data
Management Plan
Data management and data analysis, two key components of the monitoring study that
are often overlooked in the design of monitoring programs, are as important to the
success of the monitoring effort as the collection and laboratory analysis of field data.
Moreover, the cost of effective data management/data analysis can be substantial. On
the order of 20 percent of the budget allocated for the monitoring program should be
reserved for data management and data analysis activities. Failure to plan for these
costs can result in the loss of information due to inadequate data preservation and
limited analysis of the monitoring data that are collected.
DESIGN AND "
IMPLEMENT DATA
MANAGEMENT PLAN
Recent characterization efforts conducted by individual estuary programs, at consider-
able expense, found that historical data are not readily available and that essential
quality assurance information necessary to evaluate the comparability of data sets is
often not preserved. Generally, it was found that although large expenditures are often
made on data collection, the amount of funding allocated to data management and data
analysis is relatively small and inadequate.
-------
The need to assimilate and integrate historical information, as well as planned monitor- 6.1
ing efforts, should drive the development of a data management strategy. Failure to Data Management
plan for data management can result in the loss of the information due to inadequate
data preservation.
The development of a data management strategy must consider the following ques-
tions:
• Where will the data go?
• How will these data be stored?
• Who will maintain the data base?
• How will data be checked and loaded into the data base?
• How accessible will the data be?
• Will statistical, graphical, and report generating tools be available?
• How much will it cost?
A computer system will be essential for the management of the data collected by the
estuary monitoring programs. It should be operational prior to implementation of the
monitoring program and should have the following attributes:
• Centralized storage of raw data
• Easy access and use
• System documentation
• Quality assurance procedures
• Linkage to graphical, statistical and report generation routines
• Long term availability and flexibility
A centralized data source provides the ability to transfer data between investigators
and to conduct analyses that utilize data from different monitoring program compo-
-------
53
nents. An additional feature of a centralized data management approach is the ability
to designate a system administrator who has responsibility for system documentation
and data quality assurance.
The quality assurance information that must be reported with each data set must be
defined prior to implementation of the monitoring effort. The objective is to identify
key field, laboratory and quality assurance information that would allow future users
of the data to make informed decisions regarding the comparability of historical data
sets. This set of basic reporting requirements should be developed for all data types
collected. The Ocean Data Evaluation System (ODES), supported by EPA's Office of
Water, and the data system developed for the Puget Sound Ambient Monitoring
Program provide good models for the implementation of data quality assurance
procedures. Each data set that is submitted to ODES, for example, undergoes an
extensive quality assurance review, and the results of this review are summarized in a
report that is accessible from within the system. Extensive documentation for ODES
is available (U.S. EPA, 1987d and 1988b).
The data management system that is adopted by the estuary programs must also
provide basic graphical, statistical and report generation capabilities and/or the ability
to download data easily to data analysis packages. It must also be flexible enough to
address new data types and analytical needs. Finally, a long term financial commit-
ment must be made to the system in order to establish user confidence in the ability to
store and access data over a long time period.
The NEP has developed a data management policy to ensure that all potential users,
both inside and outside the NEP, have access to environmental data generated under
the program. Responsibility for identifying and selecting data management support
remains with the NEP Management Conferences. The selection of a data management
system by each program should be based on an evaluation of characterization and
monitoring requirements for the estuary. Use of existing systems, where possible, is
encouraged.
6.2
Data Analysis
As indicated in Section 5.0, an essential element of the monitoring plan will be the
specification of a timetable for analyzing the data and assessing monitoring program
performance. The assessment of monitoring program performance should be used to
refine monitoring program objectives and modify individual monitoring program
elements to satisfy these objectives. Initially, monitoring program evaluations should
be conducted after the first year of data collection. Subsequent interim evaluations
should be conducted at two or three year intervals.
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54
The primary purpose of other data analysis activities will be to test the hypotheses
developed in Step 2 of the design process (see Section 3.0). Additional goals are to
summarize the data, generate new hypotheses, and evaluate the uncertainty associated
with the measurements and conclusions. Additional analyses should be designed to
produce information for use by groups with diverse technical backgrounds.
A wide range of statistical and graphical tools are readily available for use in meeting
these goals. Recently, there has been an increased interest in the development of
Geographical Information System applications for graphical analysis and information
display. The National Estuary Program is currently funding projects to demonstrate
the use of this tool to synthesize a broad range of data collected by the estuary pro-
grams, and to effectively communicate this information to interested groups.
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55
..v-^
7.0 Communicate Program Results
One of the primary goals of the monitoring program is to provide information that can
be used to redirect and refocus the CCMP. To achieve this goal, emphasis should be
placed on distributing the data that are collected in the estuary monitoring programs.
The data collected by individual investigators should be made readily available to the
scientific community for comparative studies that relate information from different
components of the program. Section 6.0 discusses the need for the development and
implementation of an effective data management strategy. Emphasis should also be
placed on the analysis of these data. The dissemination of the recorded monitoring
data is not a sufficient mechanism for communicating the monitoring results. Statisti-
-------
cal analysis of the monitoring data is essential, and graphical and written summaries
should be produced for agency managers charged with implementing the CCMP. The
results must be effectively communicated to an audience with a wide range of techni-
cal backgrounds and interests.
Figure 1-1 and the schematic above show two feedback loops associated with the
evaluation of monitoring data. One provides direct feedback of analytical results that
are used to modify and refine the monitoring program to increase efficiency (see
Section 5.0). The other provides feedback to three basic factors that influence the
design, development and refinement of monitoring program objectives: public con-
cerns, modeling, and research. Data analyses must provide information that addresses
the needs of program managers, scientists and the public.
Graphical and written summaries should be produced that demonstrate the results of
individual components of the monitoring program as weU as the relationships between
monitoring activities. These summaries should serve as tools to effectively communi-
cate information on the effectiveness of the actions taken under the management plan,
and to build public awareness of actions taken by the estuary program. Demonstration
materials that summarize program results should be produced for use in newsletters,
workshops, poster sessions, and public forums. The results of the monitoring effort
and the data analyses should also be made available to the scientific community, and
use of the monitoring data should be encouraged. Data analyses should be conducted
to test for trends, test and generate new hypotheses, evaluate the uncertainties associ-
ated with the data, and to identify the source of these uncertainties. These analyses
should serve as a basis for extending existing knowledge of the estuary, making
refinements to conceptual and numerical models of the system developed in the
characterization phase of the program, and identifying new research. Collectively the
analytical results should provide the necessary information for redirecting and
refocusing the CCMP.
There are several examples of publications by estuary programs to generate public
interest and support, and to disseminate information on monitoring results. In addition
to preparing annual technical reports on monitoring efforts, the Puget Sound Estuary
Program produces a quarterly newsletter - Puget Sound Notes (Figure 7-1). This
technical newsletter has a distribution of 2,750 individuals and organizations. It is
intended to report on recent program results and to inform interested individuals about
events that affect the estuary. The Chesapeake Bay Barometer is a one page monthly
publication of the Chesapeake Bay Program. The Barometer provides a summary of
dissolved oxygen concentration and water clarity in the bay during the previous month.
It also includes short, nontechnical summaries of topics of general interest (Figure
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57
7-2). Previous topics have included: salinity in the bay, boat pollution, bald eagle
populations, and the striped bass fishery. The Santa Monica Bay Restoration Project is
producing a series of paired reports on pathogens (Figure 7-3). These reports summa-
rize the results of the long term assessment of inputs of fecal indicators and enteric
viruses from storm drains. The results of each phase of the study are summarized in
both a Technical Report and a Public Summary.
Figure 7-1. Sample cover
o/Puget Sound Notes.
Sound
Number 25 -June. 1991
Editor: Timothy W. Ransom, PSWQA
Contributors: Timothy W. Rtraom, PSWQA
Rontld U. Thorn, BittiUt Mlrtnt SC/WICM Laboratory
LoAnn Htllum, FltturtM Rmarch Institute
Chirln A. Slmmitid, Fltntrtn Htmreh Inrttuu
Curt/j D. Tmim, Port o/Suftfe
Fnd Wtlnmtnn and Hlchul Rylko, U.S. EPA-Rtalon W
Robf ra Fains, fSWOA
The Estuarine Habitat Assessment Protocol
oyCharlnA. SmtntUd. Wetland ecosystem Team, Fisheries Research Institute, University ol Washington, CurttD. Tinntr, PonolSeattle. aniJFnd
mlnminn ana Michael Rylko. U.S. Environmental Protection Agency-Region 10, Wetland Program and Olflca ol Coastal Waters.
Introduction
Presently, our ability to manage and preserve wetland and
other habitats of the waters of the United States is inhibited to
a large degree by the lack of consistent assessment and
monitoring. This is particularly true In the Pacific Northwest.
which is relatively young in terms of the scientific information
base. Net loss of wetlands continues in this region, even within
the regulatory domain of the CWA Section 404 permit process
(Kentula et al., in revision; Rylko and Storm, in prep.). This loss
Is largely due to a lack of consistency In the assessment and
monitoring requiredto determine which wetland functions will be
lost to a development project and thus the compensation
actions to be taken. Furthermore, this lack of consistency
impairs our ability to determine quantitatively if lost functions
have been fully replaced by the mitigation action. We argue that
one reason beWnd our inability to compel functional
replacement Is the lack of a uniformly acceptable assessment
procedure that developers and their consultants and permrtting
and resource agencies can mutually embrace. For Instance, ol
73 Section 404 permits issued between 1980 and 1990, the fish
and wildlife habitat function of the site to be filled was evaluated
for only 11 %, and monitoring of the mitigation site was required
for only 53% (M. Rylko, pers. comm.). Those proposing wetland
mitigation are hampered by the lack of any guidelines for
assessing the potential wetland Impacts of their project, and the
regulating agencies are equally hindered by the Inconsistency
and inapproprlateness ol the proposed assessment and
monitoring plans.
rocedures available, but the broad consensus among
unique wetland functions in specific geographic regions and are
typically too subjective to provide consistent results. To
Increase the effectiveness of our management and
conservation of estuarlne habitats, we need assessment and
monitoring procedures that: (1) are based explicitly on habitat
function; (2) are specific to the region of application; (3) use
methods that are standardized, consistent and comparable; (4J
generate quantitative data rather than qualitative indices; (5)
are designed to be thoroughly objective among different users
and sites; (6) will be adaptive In terms of building on prior results;
and (7) enable the user to measure wetland function from both
a single species and multi-species (community) levels.
The Estuarine Habitat Assessment Protocol ("Hie
Protocol"; Slmenstad et al., in press) represents such an
approach to assessing the function of estuanne habitats forflsh
and wildlife. Fish and wildlife support functions of estuarlne
habitats were the chosen focus of this Protocol because they
have historically been the driving criteria behind resource
agency requirements for compensatory mitigation. However,
other, potentially more important habitat functions, such as
maintenance of water quality or flood desynchronlzation, also
need to be assessed with similar rigor. The Protocol is intended
to address the need for a systematic procedure that can be
applied uniformly across a variety of wetland and associated
nearshore habitats, using objective, scientific methods.
Although It is directly applicable to the evaluation of
compensatory mitigation projects in estuarine habitats, In the
long term the Protocol may facilitate the development of
successful design criteria for estuarine habitat restoration as
well.
Concept, Development, Content and Limitations of
the Protocol
Concept
There are two fundamental problems with previous methods
of assessing estuarlne habitats: (1) documentation of simple
absence or presence of fish and wildlife, or even more
quantitative information on population sizes, in aspeciflc habitat
or at a specific site has not necessarily Indicated utilization per
sa; and (2) the causal association between characteristics of the
habitat and the function and extent of fish and wildlife utilization
has neither been Identified nor quantified. For example, while
unique nesting of wetland birds Is obviously illustrative of a
functional dependence upon the habitat, the occurrence of
highly mobile animals such as migrating Juvenile salmon does
not directly Infer functional dependence.
The Protocol differs from most prior approaches to estuarine
habitat assessment and monitoring forfish and wildlife because
it focuses on the attributes of the habitats that promote fish and
wildlife utilization. The strategy for developing the Protocol was
one of identifying the biological and physical attributes of
estuarine wetlands which determine the extent of fish and
wildlife utilization. In addition, our approach to gathering the
information for the Protocol was to identify those attributes
Important to the greatest number of fish and wildlife species.
Thus, an assessment of these attributes is presumed to
address a broad scope of the habitat's biotic community, rather
than just a single target species. It then follows that the
incorporation of such critical attributes in a wetland mitigation
and restoration design would increase the function of the habitat
for the representative community.
Development
The concept of the Protocol emerged from the deliberations
of an ad hoc group called the Urbanized Estuary Mitigation
Working Group (UEMWG),atechnicalgathering of government
agency, tribal, university and industry representatives which
has met since 1986 to discuss mitigation In urbanized estuaries.
After the basic concept of the Protocol and a process by which
it could be developed were generated by the UEMWG, funding
was assumed by the U.S. Environmental Protection Agency,
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58
"^
CHESAPEAKE BAY BAROMETER
ENVIRONMENTAL CHARACTERISTICS OF THE BAY
DISSOLVED OXYGEN
Dtvetotf o«r|» iDOl u iF« MnooM of oijftm
*MCf f*Ktt* *svr •Mdlty hu • PTMH (mow* of OO t
•ivt MM tf« bonom dw IB M§ mencuM *tdi the
**4 •ijtoi (rrtatiitfi frr >*•"« pNJpi)TxJKM. DO knta M
Umft *d tho pnbkmi that begsn »
.(ocw4 twde* IM nw Mri «MiMic!r U» Bay. Pvuculartr M UM lowtr bum.
A
^Sfea
Figure 7-2. Sample cover
of Chesapeake Bay
Barometer.
Figure 7-3. Sample
covers of Santa Monica
Bay Restoration Project
Reports.
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59
t s<"s' S i" "•%
.,:;•.•. i","--,
% ^ •, «•. .• •.
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i&f X #• X A X
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63
's ••
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64
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Appendix A - Case Studies
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A-3
A1.0 The Puget Sound Ambient Monitoring
Program Case Study*
A1.1
Purpose and
Approach
The Puget Sound Ambient Monitoring Program (PSAMP) is a comprehensive moni-
toring program which examines environmental variables throughout Puget Sound and
the surrounding watersheds. With the designation of Puget Sound as an estuary of
national significance in 1988, PSAMP was adopted and submitted as part of the
designation package of the National Estuary Program. The development, implementa-
tion, management, and coordination of PSAMP can serve as an excellent model for
estuarine monitoring programs nationwide.
The sequence of events leading to the implementation of the Puget Sound Ambient
Monitoring Program is shown in Figure Al-1. Important features of this time line are
that the development of this comprehensive monitoring plan was well underway prior
to the selection of Puget Sound for the National Estuary Program, and that the devel-
opment and implementation of such a comprehensive monitoring program takes
substantial time. The PSAMP design was developed between 1986 and 1988, and
sampling began in 1989. However, the Puget Sound Water Quality Authority
(PSWQA), which played a key role in its development, was formed in 1983 and issued
a report in 1984 which recognized the need for estuary-wide monitoring.
Throughout the process of developing PSAMP, National Estuary Program (NEP)
objectives were taken into account. PSAMP fulfills the monitoring requirements of
the NEP, including:
• identify trends in water quality, natural resources, and uses of the estuary,
• develop the relationship between loads and potential uses,
• identify causes of environmental problems, and
• monitor the effectiveness of actions taken under the Puget Sound Water
Quality Management Plan (CCMP).
This case study describes the steps involved in the development of PSAMP, the goals
of the program, the monitoring design, and the relationship of PSAMP to the Compre-
hensive Conservation and Management Plan (CCMP) for Puget Sound. Emphasis is
placed on describing the institutional aspects of the monitoring program development
process, including the roles of the participating federal, state, local, and tribal govern-
ments. The importance of scientific review and input from the public is also empha-
sized.
*This case study was adapted from a paper prepared by Andrea E. Copping of the Puget Sound Water
Quality Authority and John W. Armstrong of the U.S. Environmental Protection Agency, Region 10, for
the January 1991 Puget Sound Research Conference.
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A-4
Figure Al-1. Events
leading to the development
and implementation of the
Puget Sound Ambient
Monitoring Program.
1987
- Puget Sound Water Quality Authority (PSWQA) established
- PSWQA Annual Report recommended creating a Sound-wide Water Quality
monitoring program
- Legislation revised; PSWQA required to prepare and adopt a Puget Sound Water
Quality Management Plan and to include recommendations for a comprehensive
monitoring plan
- PSWQA released Issue Paper on comprehensive monitoring
- EPA Region X released a monitoring strategy for a Sound-wide monitoring Program
- Puget Sound Water Quality Management Plan calls for creating an Ambient
Monitoring Program and creates the Monitoring Management Committee
- Public and government workshops reviewed proposed PSAMP
- Puget Sound designated in the National Estuary Program
- Monitoring Management Committee Final Report submitted to PSWQA
- PSWQA incorporated PSAMP in 1989 Management Plan
- Management framework for PSAMP adopted; PSAMP Steering Committee
established
- PSAMP data collection began
- First Annual Report of PSAM P
- PSAMP review and feedback; continued monitoring
Early in the development of PSAMP, it became clear that the cost and logistics of such A1.2
a comprehensive effort would require the participation of all key resource agencies and Development Of
affected parties. Participating local, state and federal agencies would have to make PSAMP:
modifications to existing monitoring efforts and agree to share data. It was also Institutional
Arrangements
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A-5
recognized that securing long term funding sources would require strong public
support. The key agencies and the milestones in the development and implementation
of PS AMP are described below.
The Puget Sound Water Quality Authority
In 1983 the Governor of the state of Washington appointed the 21 member Puget
Sound Water Quality Authority (PSWQA) and charged them with identifying pollu-
tion-related threats to Puget Sound marine life, evaluating pollution threats to human
health, and investigating the need for coordination among agencies responsible for
protecting Puget Sound water quality. In their 1984 report, the PSWQA called for "a
long-range coordinated plan ...to protect and improve water quality throughout the
Sound" (Booth and Powell, 1984). They further recognized the need for an estuary-
wide management plan, including increased coordination and enhancement of existing
monitoring programs.
In 1985, the Washington state legislature created the PSWQA as a state agency with a
seven member board appointed by the Governor. The enabling legislation charged the
PSWQA with developing a comprehensive management plan for Puget Sound and its
related waterways. The PSWQA was required to prepare and adopt a Puget Sound
Water Quality Management Plan by January 1987, to update the plan in 1989 and
1991, and to oversee the implementation of the plan. The legislation required that the
plan include consideration of many issues, including "...recommendations for a
comprehensive water quality and sediment monitoring program..." PSWQA was also
charged with creating a biennial "State of the Sound" report to describe water quality
and related resource conditions in Puget Sound. Throughout its history the PSWQA
has relied heavily on input from all parties affected by water quality, regulatory
frameworks, and the need to manage the resources of Puget Sound.
PSWQA Monitoring Issue Paper
As part of the development of the 1987 Puget Sound Water Quality Management Plan,
the PSWQA wrote an issue paper on Comprehensive Monitoring of Puget Sound
(PSWQA, 1986). The issue paper reviewed existing Puget Sound monitoring pro-
grams, described the process for developing a comprehensive water quality and
sediment monitoring program which includes biological resources, and presented
alternatives for developing the monitoring program (Table Al-1). The issue paper
identified four major components of a comprehensive monitoring program:
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A-6
. ^
5" Sȣ
TABLEAU. ALTERNATIVES
FOR IMPROVING PUGET SOUND MONITORING
Development and adoption of standardized protocols
A centralized or coordinated data management system
Improved data mterpretation/reportpreparation and information dissemination
Adequate laboratory capacity s '
Review of existing water column program and changes to collect more pertinent
information. Follow up actions could be tied to monitoring results.
Development of a sediment monitoring program, including ambient sediment moni-
toring, intensive surveys, discharge monitoring, and dredging/dredged material
disposal monitoring , „,,.. , „ , ', „
Improved point and nonpoint source monitoring for municipal treatment plants and
Industrial facilities, searches for illegal discharges
Greater number of intensive surveys to identify sources of pollution
Development of a biological monitoring program
Improved monitoring of commercial and recreational shellfish beds
Monitoring by citizen volunteers
Reference: PSWQA1986.
• ambient monitoring
• discharge monitoring/compliance monitoring
• intensive surveys
* resource monitoring and other programs (e.g., monitoring paralytic shellfish
poisoning, changes in shoreline and habitat, weather, demographics, and
socioeconomic information.)
The PSWQA included findings of the 1986 issue paper, and comments received on the
issue paper, in the 1987 Puget Sound Water Quality Management Plan (PSWQA, 1987).
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EPA Region 10 Monitoring Guidance Report
In 1986, the Office of Puget Sound, EPA Region 10, directed their consultant to
develop a monitoring strategy for a Sound-wide monitoring program. Through a
process involving consultation with agencies concerned with Puget Sound manage-
ment, the consultant developed a monitoring design which included goals and objec-
tives, sampling design, operation of the program, and methods for sampling, analysis,
and reporting of data. The monitoring design was released in draft form in 1986 (Tetra
Tech, 1986). At that point EPA Region 10 and the PSWQA recognized the need for a
process which would further involve the many agencies, organizations, and individuals
with an interest in Puget Sound monitoring.
Monitoring Management Committee
The 1987 plan required the PSWQA to appoint a Monitoring Management Committee
(MMQ (PSWQA, 1987). The MMC is chaired by the PSWQA and consists of
representatives of state, federal, local, and tribal governments, business, industry,
shellfish growers, university scientists, Canadian agencies, environmental groups, and
the public. The plan directed the MMC to develop a comprehensive ambient monitor-
ing program, and to address 16 specific issues related to the program (Table Al-2).
Staff support was provided to the MMC to carry out coordination and data manage-
ment tasks.
The MMC was appointed in September 1986. Like all PSWQA meetings, MMC
meetings are open to the public. The committee reaches all decisions by consensus.
The committee began its work in late 1986 by defining the goals and objectives of the
monitoring program and then proceeded to review and modify the consultant monitor-
ing strategy (Tetra Tech, 1986). The MMC established technical subcommittees to
study the monitoring design and to make recommendations on program design.
From late 1986 to early 1988 the MMC refined the monitoring program design which
included sampling and analysis recommendations, a data management system design,
quality assurance guidelines, reporting requirements, cost estimates, and funding
alternatives. The committee released a draft program design in May 1987 for public
and scientific peer review. Workshops were held for the public, local government, and
tribal staff. Comments received from the PSEP technical advisory committee (TAG),
scientists, agency staff, and other affected parties were considered by the committee
and a final report entitled "Puget Sound Ambient Monitoring Program - Detailed
Design Considerations" was adopted by the PSWQA and published in April 1988
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A-8
TABLE Al-2. MMC ACTIVITIES REQUIRED B* PSWQA
IN DEVELOPING PS AMP
• Define the goals and objectives of the monitoring program.
» Define how the monitoring data will be used to assist in management of Puget Sound
resources by responsible agencies,
« Review and provide for the specific monitoring needs of programs in thePaget Sound
Water Quality Management Plan.
* Propose a monitoring design based to the extent applicable on the November 1986
consultant report to EPA on Puget Sound monitoring and review a similar report on
freshwater (watershed) monitoring.
* Analyze the data needed to meet the objectives of the monitoring program,
• Analyze and evaluate existing programs and their relation to the objectives of a
monitoring program. " '"""" " -
• Identify comprehensive monitoring needs not being met by existing programs.
• Identify mechanisms to obtain needed information either through redirection of
existing programs Orinitiating of new programs. ,,/"
• Estimate annual costs associated with existing monitoringMPugetSouhdandtcosts of
additional needed monitoring.
* Recommend a data management system that meets agency needs.
• Define appropriate mechanisms to fund the monitoring program including allocation
of existing funds and new funding.
* Define the appropriate roles for agencies In implementing the monitoring program.
• Define how the Puget Sound Atlas (Evans-Hamilton and DIET Systems 1987) would
be updated, managed, and integrated with ongoing activities*
• Recommend a structure for managing the monitoring program, including mechanisms
for making needed refinements and adjustments to ensure program objectives are met
in a cost-effective mariner. - ' ' ~-", -
• Identify opportunities for citizens' monitoring of Puget Sound,
* Form ad hoc technical working groups to assist in developing the monitoring program.
Reference: PSWQA, 1987.
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A-9
(PSWQA, 1988a). The report included PSAMP goals and objectives, and sampling
and analysis strategies for monitoring sediment quality, marine and fresh water quality,
fish, shellfish, birds, marine mammals, river mouths, nearshore habitat, and additional
data. The PSAMP design was incorporated by reference into the 1989 Puget Sound
Water Quality Management Plan (PSWQA, 1988b).
A1.3
The Puget Sound
Ambient
Monitoring
Program
The MMC developed five goals for PSAMP:
1) Characterize and interpret spatial and temporal patterns of conditions of
Puget Sound in relation to its natural resources and for humans, and
recognize contamination.
2) Take measurements to support specific program elements identified in the
Puget Sound Water Quality Management Plan.
3) Measure the success of programs implemented under the Puget Sound
Water Quality Management Plan (as they relate to the overall ambient
monitoring goal and the program goals of the plan).
4) Provide a permanent record of significant natural and human-caused
changes in key environmental indicators in Puget Sound over time.
5) Support research activities through the availability of consistent, scientifi-
cally valid data.
In order to achieve these goals, the monitoring program characterizes and interprets
spatial and temporal patterns for the following:
• factors that endanger human health
• biological populations and communities
• factors affecting biological populations
• presence of pollutants in the Sound
• entry of pollutants into the Sound from rivers and streams
• estuarine and wetland habitats
• results of water quality degradation, such as shellfish bed closures
• improvements in water quality such as the reopening of shellfish beds
• factors that affect aesthetic conditions.
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A-10
The monitoring program identifies areas where:
* resources are damaged or contaminated
• resources or conditions are changing over time
• intermittent or periodic degradations are occurring.
The Scope of PS AMP
The design of PSAMP calls for monitoring of sediments, water quality in marine and
fresh waters, fish, shellfish, birds, marine mammals, and nearshore habitat. The focus
of the program is Puget Sound-wide, which includes Hood Canal, portions of the
Straits of Juan du Fuca and Georgia to the Canadian border, and the surrounding
watersheds. The sediment quality sampling stations, for example, are shown in
Figure Al-2. Sampling occurs year-round. Station locations, the number of stations
or surveys, and the timing of sampling varies with each type of measurement. Three
types of stations are sampled in many tasks of the program:
• fixed stations that are sampled each year,
• rotating stations that are sampled on a 3 year cycle; these stations can
provide extended geographic coverage, and
* a small number of floating stations that are located at the discretion of the
implementing agency; these stations can provide additional geographic
coverage, or allow investigation of a suspected water or sediment quality
problem.
Table Al-3 summarizes the design for full PSAMP implementation, as well as the
monitoring data that are being collected during 1989-90.
Citizens' monitoring is an integral part of PSAMP. The advantages of using citizens
to collect data are twofold: samples and observations can be collected in a cost-
effective manner, particularly at remote locations where travel by agency staff is
costly; and as a tool for increased public involvement, education, and stewardship.
Using state funds, PSAMP contracts with citizens' groups to provide volunteers to
collect information and samples under the guidance of implementing agency staff
(funds are used for administrative purposes and to reimburse expenses). Parameters
for collection by citizen monitors are carefully chosen to minimize quality assurance
problems; for example, citizens are used extensively to dig shellfish for tissue analysis,
to catch marine fish and salmon for tissue analysis, and to groundtruth remote sensing
data.
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A-11
Figure Al -2. Area
included in Puget Sound
Ambient Monitoring
Program.
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A-12
TABLE Al-3. PSAMP DESIGN AND 1989-90 ACTIVITIES
Task
Se-dlmcnt quality
Marine water
column
Rsh
Shellfish
Birds
Marine mammals
Nearshorchabitai
Freshwater
Task
Subcomponents
Scdimentchemistry
Bioassays
Benthicinvertebrates
Long-term trends
Known water quality "
problems
Algal growth
Tissue chemistry
(bottomfish)
Liver histopathology
(bottomfish)
Tissue chemistry
(cod, rockfish, salmon)
Abundance
Bacterial contamination
Tissue chemistry
Paralytic shellfish
poisoning
-
•,
Proposed Actual
No. of , 1988-89
Stations/Surveys Stations/Surveys
75 throughout
Puget Sound
10-12 throughout'
Puget Sound
5-10 in selected bays
5-10 in selected bays
21 stations'
21 stations
5-10 stations
35 beaches
35 beaches
35 beaches
35 beaches
.Throughout
Puget Sound
Throughout
Puget Sound
One-third of
Puget Sound
75 throughout
watershed
50 throughout
Puget Sound
24 throughout
Puget Sound
10 stations
10 stations
4 stations
10 beaches
lObeaches
4 beaches
16 beaches
No activity
No activity
No activity
Proposed
Sampling
Frequency
AnnnjaUy-
spring
Monthly
Annually-
•• ' early summer
Annually-
early summer
Annually,
depending
on species
Annually
Quarterly
Annually
Monthly
Monthly, annually
Monthly, annually
Annually
75 throughout Monthly
watersheds
(limitedparameters)
.Actual
Sampling
Frequency
March-April
Monthly
Seasonally
Stuamer/wintef
solstices
Mayl989
May 1989
September/
February/
April 1989-1990
May 1990
Quarterly
April 1990
Monthly
Monthly
Reference: PSWQA1990a. J/- ~ -
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A-13
The NEP and PSAMP
With the formal designation of Puget Sound as an estuary of National Significance in
March 1988, the 1991 Puget Sound Water Quality Management Plan was adopted as
the Comprehensive Conservation and Management Plan, and PSAMP formally
became a part of Puget Sound Estuary Program.
PSAMP examines data from selected other monitoring programs and studies underway
in the Puget Sound basin in order to better interpret PSAMP data, and to meet the
needs of the NEP which states that a CCMP "shall survey and utilize existing reports,
data, and studies relating to the estuary that have been developed by or made available
to federal, interstate, state, and local agencies." Monitoring programs and studies that
evaluate environmental conditions and potential sources in the Puget Sound region that
are of interest to PSAMP include:
• nonpoint source control watershed monitoring
• Puget Sound Dredge Disposal Analysis (PSDDA)
• PSEP Urban Bay Studies (UBATs)
• NPDES compliance monitoring
• Department of Ecology intensive surveys (tracing sources of contamina-
tion)
• NOAA National Status and Trends Program
• research studies, historical information, and other studies.
• Timber/Fish/Wildlife
• climate/weather data
• demographic and socioeconomic conditions
• decision record-keeping.
A1.4
Implementation of
PSAMP and Cost
Assuring PSAMP Implementation
In order to ensure that PSAMP would be implemented as planned, the PSWQA
negotiated memoranda of agreement (MO A) with each of the implementing agencies.
These agreements detailed the specific responsibilities of each implementing agency
and the PSWQA in relation to PSAMP. The MOAs specify that each implementing
agency must carry out its responsibilities under the PSAMP design, must maintain
PSAMP funding levels within the agency unless the Governor or the state legislature
requires across-the-board agency funding cuts, and must refer changes in PSAMP to
the PSAMP Steering Committee for approval. Through the MOAs, the PSWQA
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A-14
agrees to provide staff support to the PS AMP committees and to work with the imple-
menting agencies to maintain long-term funding for PSAMP.
Under the 1985 legislation creating the PSWQA, the agency was due to terminate in
1991. In order to ensure the future of PSAMP, during fall 1989 the PSAMP Steering
Committee developed a recommendation for future management structure for PSAMP
which was not markedly different from the existing one. The PSAMP Steering
Committee preferred that the coordination and dispute resolution functions remain
with the PSWQA. The committee's recommendation was incorporated into the
PSWQA enabling legislation in 1990, which reauthorized the PSWQA. The new
section in the legislation which deals with PSAMP requires the implementing agencies
to participate in the program, requires the PSWQA to ensure implementation of
PSAMP, and allows the PSWQA to establish an interagency committee to coordinate
PSAMP.
Initial PSAMP Assignments
Agencies represented on the MMC were asked to express their interest in playing a
lead role in specific tasks of PSAMP. "Implementing agencies" were chosen for each
PSAMP task based on these expressions of interest. It came as no surprise that the
agencies that expressed interest in becoming implementing agencies were those with
regulatory and/or management responsibilities as well as expertise in the specific
areas; initially all implementing agencies are state agencies (Table Al-4), although this
could change in the future. Each implementing agency was responsible for preparing a
detailed implementation plan, managing their specific PSAMP task, quality assurance
and quality control, data management and reporting, and transfer of PSAMP data to
the central database, housed at the PSWQA. The MMC felt strongly that the PSWQA
should continue to play the lead role in coordinating PSAMP and providing data
management functions, as the Authority has both the expertise and staff available to
carry out these functions.
Cost
A technical subcommittee to the MMC (Costing Subcommittee), PSWQA staff and a
consultant retained by PSWQA worked to develop a cost estimate for PSAMP. In
1987 dollars, full implementation of PSAMP was calculated to cost approximately
$3.2 million a year, with startup costs of $250,000 to $350,000 over the first two years.
Early in 1988 it became apparent that state funds would not be available for full
implementation of the program. Through a priority-setting process involving the
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A-15
TABLE Al-4. PS AMP TASK ASSIGNMENTS BY AGENCY
Monitoring Task
Implementing Agency
Sediment Quality
Marine Water Column
Fish
Shellfish
Birds:
Marine Mammals
Nearshore Habitat
Freshwater
Reference: PSWQA I990b
Washington Department of Ecology
Washington Department of Ecology
Washington Department of Fi$heries
Washington Department of Health
Washington Department of Wildlife
Washington Department of Wildlife
Washington Department of Natural Resources
Washington Department of Ecology '
MMC and other scientists, a plan for phased implementation of the program was
devised. This process established the order in which PSAMP tasks would be funded
when implementation funding became available. The MMC established sediment
quality, fish tissue, and shellfish tissue monitoring as top-ranked priorities for funding.
The MMC also recognized that agency staff could not manage to fully implement
PSAMP in a single year, even if funds were to become available.
The development phase of PSAMP, including establishment of the data management
system, has been funded largely by PSEP funds administered by EPA Region 10 and
made available to the PSWQA by cooperative agreement. State, federal, local, and
tribal agencies, as well as private sector funds have contributed individuals' time
serving on monitoring committees and subcommittees. The development costs of
PSAMP were approximately $200,000 from 1987-1989. Additional development
costs for data management are still being incurred.
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A-16
•'X^sJlS
Funding
Long-term stable funds for monitoring programs are notoriously difficult to secure.
Funding for four types of activities were needed to assure the success of PSAMP:
contractor funds for the development of the draft monitoring strategy and several
workshops; staff time for program development; startup costs; and ongoing implemen-
tation. As previously noted, EPA Region 10 provided PSEP funds for PSWQA staff to
coordinate the development of PSAMP and the data management system. PSEP
resources continue to fund PSWQA staff for initial implementation and further data-
base development through FY91. Start up costs for implementing PSAMP have been
itemized together with long-term implementation costs to clarify what is needed to
properly initiate such a program.
To date, approximately 30% of PSAMP has been funded, including central coordina-
tion and data management tasks. Other than PSEP funds available through EPA, all
PSAMP funding has come from the state of Washington. Like all Puget Sound Water
Quality Management Plan funds, PSAMP funds are given directly to the implementing
agency; the PSWQA submits a funding request for the entire plan, with assistance
from the other agencies, and acts as an advocate for the plan to the Governor and the
state legislature.
During 1989-90, PSAMP received approximately $900,000 of state general fund
money each year (PSWQA 1990a). The Washington state Department of Ecology
receives $500,000 a year for sediment sampling and analysis. $200,000 goes to the
Washington state Department of Fisheries for measuring toxics in bottomfish and
recreational fish tissue and for bottomfish health assessments. The Washington state
Department of Health uses $160,000 for sampling and analysis of shellfish tissue for
bacterial and/or chemical contamination. The PSWQA receives $40,000 a year for
coordination and data management. $50,000 in citizens' monitoring activities arc
funded by a special fund for public involvement and education (PIE Fund) from the
state Centennial Clean Water Fund (tax on tobacco products). In addition to securing
new funds, the Washington state Department of Ecology has modified existing marine
and fresh water monitoring programs to meet PSAMP goals, saving PSAMP $245,000
a year.
Maintaining current PSAMP funding and securing additional funds is a major focus of
the PSAMP Steering Committee and the PSWQA. The MOAs are intended to protect
PSAMP funds within the implementing agencies, all of which have numerous conflict-
ing priorities for funds. The monitoring committees and the PSWQA are relying
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A-17
heavily on legislative advocacy, attention from the media, and garnering of public
support through the dissemination of PSAMP results to aid in securing PSAMP funds.
Management of PSAMP
The management structure for PSAMP consists of the PSAMP Steering Committee,
the MMC and subcommittees, and the PSWQA board. PSWQA staff provide techni-
cal and administrative support to the committees, and chair the MMC and PSAMP
Steering Committee.
As PSAMP moved from the development to the implementation stage, it became
apparent that the MMC was too large and unwieldy a group to coordinate the program
and make day to day decisions. In addition, some state agencies were uncomfortable
with having representatives from the private sector make decisions involving the use
of public funds. To solve these problems, a second monitoring committee was formed,
the PSAMP Steering Committee.
The PSAMP Steering Committee is chaired by the PSWQA and is made up of one
representative from the five implementing agencies, the PSWQA, EPA Region 10,
local government, and tribes. Additional members can be added in future if necessary.
The committee is responsible for the overall coordination and management of PSAMP.
The PSAMP Steering Committee meets once a month and reaches decisions by
consensus.
Should the PSAMP Steering Committee be unable to reach consensus on technical or
other program matters, or should any member be dissatisfied with decisions made by
the committee, the matter can be referred to the PSWQA board for resolution. To
date, there have been no such instances.
The MMC maintains the same representation that it had during the development phase,
and, with its subcommittees, acts as the advisor to the PSAMP Steering Committee.
The MMC meets twice a year or as needed. Major changes in PSAMP design,
changes in assignment of an implementing agency, setting funding priorities, and other
technical issues must be referred to the MMC by the PSAMP Steering Committee.
Although the PSWQA has played the central role in facilitating the development and
initial implementation of PSAMP, the PSWQA is committed to maintaining PSAMP
as an interagency effort which benefits the public, as well as many agencies and
organizations besides the PSEP co-managers and implementing agencies.
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Review ofPSAMP
The PSAMP Steering Committee and the MMC recognize that the long-term success
of PSAMP rests on making the program effective in answering needs of the resource
management agencies, and informing the public of the state of the Sound, within the
available budget An annual review of PSAMP is built into the process by the partici-
pants in order to keep the program focussed on top priority monitoring needs and to
ensure the efficient use of funds. The first annual review will begin in late 1990. In
addition, PSAMP will undergo periodic independent review; the first review is set for
1994, with subsequent reviews every 3 years.
Data Management, Quality Assurance and Reporting
Data management and quality assurance/quality control (QA/QC) requirements for
PSAMP were designed as integral parts of the program. The data management system
consists of a central database housed at the PSWQA, and agency databases at each of
the implementing agencies. The databases are microcomputer (desktop computer)
based, and exchange data electronically using a set of standardized data transfer
formats developed by PSWQA staff in cooperation with technical subcommittees to
the MMC. The central database and several of the agency databases are on dBase IV
and have been customized by PSWQA staff.
PSAMP is also responsible for developing a geographic information system (GIS) on
Puget Sound. Resource maps from the Puget Sound Environmental Atlas (Evans-
Hamilton and D.R. Systems, 1987) have recently been converted from graphics files to
Arc-Info (GIS program) by the USGS, under an agreement with the PSWQA.
PSWQA staff are presently working with the Washington State Department of Natural
Resources to update the Atlas on GIS and will complete the initial Puget Sound GIS by
1993.
Each implementing agency is responsible for QA/QC of sample collection, analysis,
and data management for their tasks of PSAMP. Limited QA/QC of data are per-
formed at the PSWQA before data are accepted into the central database. QA/QC
guidelines were established by the MMC and are detailed in each agency's implemen-
tation plan.
Each implementing agency is required to prepare an annual technical report detailing
their PSAMP monitoring over the previous year. Using the agency technical reports
and other program information, each year PSWQA staff prepare an integrated PSAMP
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•*•; ,
report for public release. The first of these reports was released in May 1990
(PSWQA, 1990a). PSAMP reports and quality assured data are available through the
PSWQA.
A1.5
Summary and
Recommendations
This case study describes the process by which agencies, the private sector, scientists,
and the public interest groups interacted to design and implement PSAMP. It also
identifies some of the ingredients that have made it a successful process. Addition-
ally, there are several opportunities for other estuary programs to improve upon this
process.
Keys to Success
The success of PSAMP design process is attributable to the agencies and individuals
who saw the need for a comprehensive monitoring program and who made the com-
mitment to make the process work. The PSWQA played an essential role in coordi-
nating and managing the process. One key to the PSWQA's past and ongoing suc-
cesses has been the agency's comprehensive approach to complex issues by simulta-
neously addressing the scientific, resource management, political, and public con-
cerns. PSWQA staff have spent considerable effort trying to balance these often
conflicting pressures. PSWQA has been able to maintain an objective role in manag-
ing and coordinating PSAMP without competing with the individual agencies;
PSWQA does not carry out any sampling operations.
The EPA's Office of Puget Sound was very supportive of the efforts to develop
PSAMP. Not only did EPA provide funds for the development of the program, but
key staff were very much involved in the process and fully supportive of the strong
role taken by the PSWQA in coordinating the program. EPA Region 10 has sup-
ported the development of a number of other products and studies which have laid the
groundwork for PSAMP, including the Puget Sound Protocols and Guidelines; the
Puget Sound Environmental Atlas; reconnaissance studies for sediment and tissue
contamination; seafood risk assessment study; and the Puget Sound chemicals of
concern matrix.
The interest among agencies, legislators, and the public in Puget Sound water quality
during the early 1980s helped set the stage for creating the PSWQA and emphasizing
water quality issues and programs, such as PSAMP. Prior to the establishment of the
NEP, EPA Region 10 and the Washington state Department of Ecology established
the Puget Sound Action Program. These agencies were then joined by the PSWQA
and formed the PSEP.
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The design and initial implementation of PS AMP, including the data management
system, is successful both as a technical product and for the consensus process from
which it came. The involvement of representatives from both the public and private
sectors, a targeted outreach effort, and the process of peer review have all contributed
to the success with which the program has been received by the implementing agen-
cies. The data management system and data transfer formats has been very well
received by state and local agencies. PS WQA staff have had numerous requests for
information, and interest in PSAMP have been communicated from several of the
established and emerging estuary programs around the country as well as from other
state programs, other regions, and from Canada. Staff are frequently asked to present
the program design, institutional structure, data management criteria, and uses of
PSAMP data at seminars and conferences in the Puget Sound region and across the
country. In addition, PSAMP was able to release its first annual report one year after
initial sample collection.
The CIS recommendation was initially prepared to serve many purposes. Funding
constraints now and in the future have necessitated that the effort be reduced to fit the
immediate needs of updating of the Puget Sound Atlas and providing some geographic
data management for PSAMP. The flexibility of the recommendation and the broad-
based consensus through which it was built have allowed for this redirection of effort
without a loss of time or momentum.
The redirection of existing state agency funds and the procurement of new state money
for PSAMP were also aided by the long consensus process that was followed. Al-
though sufficient state funds for full implementation of PSAMP are not currently
available, almost $1 million a year is currently committed to PSAMP, and additional
resources can be expected to follow in the future.
Opportunities for increased productivity and cost containment have been seized on
several occasions during this project For example, a combination of PSEP and state
funds were used to develop the fish task implementation plan and to sample bottomfish
during 1989. Similarly, a mix of funds were used to design and develop the PSP
database. These opportunities have allowed those agencies to proceed with PSAMP
implementation without delay when state funds became available in July 1989. Simi-
larly, the planning carried out by Fisheries and the responsiveness of Fisheries and
Authority staff allowed the cost-effective "piggybacking" of bottomfish sampling with
other state sampling programs in May 1989.
The transition to a new management structure for PSAMP during the implementation
phase, including the establishment of the PSAMP Steering Committee and the reallo-
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cation of management responsibilities between the PS AMP Steering Committee and
the MMC was accomplished smoothly. The agencies involved displayed a sense of
stewardship for the program and have continued to actively participate. The success
of the management structure is most clearly illustrated in the incorporation of the
PSAMP Steering Committee's recommendation for managing PSAMP into law during
the 1990 state legislative session.
All of the implementing agencies are large complex agencies with, in some cases,
several divisions involved in the implementation. Despite the complexity of the issues
involved in long-term implementation and coordination of PSAMP, three of the five
implementing agencies have signed memoranda of agreement with the Authority.
The establishment of a successful citizens' monitoring program benefits both PSAMP
through cost-effective collection of data, and educational and public involvement
needs. Encouraging an educated and involved public is key to providing support and
commitment to the long-term funding of PSAMP.
Opportunities to Improve Future Projects
Throughout the many successes of PSAMP design and initial implementation, there
were setbacks, minor difficulties, and areas for future improvement. The majority of
these problems could be alleviated in other programs with a combination of lessons
learned from this program and the addition of more staff time.
The time to negotiate the memoranda of agreement between each implementing
agency and the Authority was seriously underestimated. The size and complexity of
several of the agencies involved in the program was not taken into account.
Questions have arisen during the design and initial implementation phases of PSAMP
as to whether it would have been preferable to design a more limited and affordable
ambient monitoring program, rather than the necessary and sufficient program de-
signed to meet the goals set out in the MMC final report (PSWQA, 1988a). A smaller
program would have been designed somewhat differently, and would not have raised
the expectations of all those eagerly awaiting PSAMP data. The design of PSAMP as
a thoroughly adequate program for characterizing Puget Sound and measuring the
success of source control and remedial action programs will act as the blueprint with
which to direct funds as they become available. The danger in designing a less com-
prehensive program is a level of complacency among funding sources and managers
who feel that we are collecting adequate information on Puget Sound. With benefit of
hindsight, we would probably design much the same program again.
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Similarly, the amount of effort that went into the design and implementation of the
central database has been questioned as little use has been made of the analytical
capabilities of the system to date. The modular design of the central database has
allowed portions of it to be customized for use by the individual implementing agen-
cies. Without the PSWQA staff effort in developing the central database, it is unlikely
that the implementing agencies would have functional databases in time to manage
their early years of PSAMP data. Additionally, as more data are collected under
PS AMP, the central database will become the site of integrated analyses among the
PSAMP tasks.
Trade offs between efficiency and the need to build broad-based consensus on all parts
of PSAMP were seen throughout the design and initial implementation process. For
example, additional portions of the central database could have been completed by the
end of the development phase if PSWQA staff had begun programming and file design
work earlier. However, an earlier start on database development would not have
allowed as complete review and participation by the implementing agencies, other
participants in PSAMP, and the interested public.
It has proved to be very important to staff the design and implementation of PSAMP
with a full-time manager/coordinator and a systems analyst. The task of keeping all
the implementing agencies and other participants informed of progress, resolving
disputes and misunderstandings, carrying out technical and administrative duties,
coordinating with other closely related monitoring and research programs, and publi-
cizing the existence and results of the program was overwhelming for PSWQA moni-
toring staff. In order to really cover all these bases, additional junior level staff was
needed. In future projects, creative use should be made of interns and entry level
professional staff, as well as additional funding sought for central coordination and
data management.
At times technical questions brought to the MMC and PSAMP Steering Committee
could not be adequately resolved due to the lack of technical expertise on the commit-
tees. Technical subcommittees were formed and proved useful for many questions;
however, inadequate PSWQA staff time did not allow for sufficient meetings and
contacts to resolve every need. In addition, there was frequently insufficient time and
attention given to the program by committee members. In some instances additional
PSWQA staff time could have improved the situation but in general there needs to be a
strong devotion to the program by some of the participants, especially the implement-
ing agencies. It is also important to recognize that there is only a finite pool of Puget
Sound scientists and managers available to participate on a regular or frequent basis in
such programs; a similar-sized pool of experts may prove limiting in many areas.
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The major activity which was given short shrift during the initial implementation of
PSAMP, and which future projects should strive to improve upon was the publicizing
of monitoring data, results, expectations, and interpretations. Even with a very limited
amount of new monitoring data there are ample historical Puget Sound data residing in
the state and federal agency files which could be resurrected, compared to recently
collected information, and displayed to managers and the public. Publicly releasing
results and interpretations of scientific findings helps to raise the level of awareness
and support for a program like PSAMP.
A1.6 Booth, P.N., and S.L. Powell. 1984. Puget Sound Water Quality Annual Report.
References Prepared for PSWQA, Olympia, WA. 39 pp.
Evans-Hamilton and D.R. Systems. 1987. Puget Sound Environmental Atlas. Pre-
pared for U.S. Environmental Protection Agency, Puget Sound Water Quality Author-
ity, and U.S. Army Corps of Engineers, Seattle, WA.
PSWQA. 1986. Issue Paper: Comprehensive Monitoring of Puget Sound. PSWQA,
Seattle, WA. 34 pp. + appendices.
PSWQA. 1987. 1987 Puget Sound Water Quality Management Plan. PSWQA,
Seattle, WA.
PSWQA. 1988a. Puget Sound Ambient Monitoring Program - Detailed Design
Considerations. Monitoring Management Committee Final Report. PSWQA, Seattle,
WA. 145 pp.
PSWQA. 1988b. 1989 Puget Sound Water Quality Management Plan. PSWQA,
Seattle, WA. 276 pp.
PSWQA. 1990a. Puget Sound Update: First Annual Report of the Puget Sound
Ambient Monitoring Program. PSWQA, Seattle, WA. 89 pp.
PSWQA. 1990b. Draft 1991 Puget Sound Water Quality Management Plan. PSWQA,
Seattle, WA. 320pp.
Tetra Tech, Inc. 1986. Puget Sound Monitoring Program: A Proposed Plan. Draft
Report Prepared for U.S. Environmental Protection Agency, Seattle, WA. 103 pp.
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\
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A2.0 Chesapeake Bay Monitoring Program:
Detection of Trends in Estuaries
A2.1 This case study demonstrates the use of statistical methods to evaluate the ability of
Purpose ongoing monitoring efforts to detect long term trends in the estuary. As described in
and Approach Section 5.0, this type of evaluation can be used to determine if the overall goals of the
monitoring program are being met and if the existing program should be modified.
Water quality data collected as part of the Chesapeake Bay Monitoring Program were
obtained to demonstrate the analytical approach. However, this case study is not
intended as a review of the ongoing monitoring efforts in the Chesapeake Bay. The
data that were used represent a small subset of the data that have been collected as part
of a comprehensive sampling effort. The ongoing monitoring program also has many
objectives that are not addressed in this example analysis.
Background information on the Chesapeake Bay Program is included, and the devel-
opment and implementation of the nutrient reduction strategy that is a cornerstone of
the bay-wide management plan is described. A description of the water quality data
that were obtained from the Chesapeake Bay Program is also included. The focus of
the case study, however, is on describing the use of statistical analyses to evaluate
monitoring effectiveness and the application of the analytical results. An understand-
ing of statistical power analysis is necessary to fully understand the procedures de-
scribed. Familiarity with the information provided in section 5.0 is therefore recom-
mended.
A2.2
Chesapeake Bay
Program
The Chesapeake Bay Program, first authorized by Congress in 1977, has developed a
"long-term process of collecting critical environmental data throughout a 66,500
square mile ecosystem" (Chesapeake Bay Program Monitoring Subcommittee, 1989).
Due to the size of the bay, a cooperative effort between numerous state and federal
agencies was required to build the current program. Efforts during 1976 through 1983
were spent developing a program and communication structure.
Early characterization efforts indicated that:
• The bay was over-enriched with nutrients
• Toxic "hot spots" exist
• Submerged aquatic vegetation habitats had been lost
• Lowered dissolved oxygen levels were causing serious water quality
problems and the loss of valuable habitats
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A-26
^ •. f -yiy-fssfff fl
In particular, increases in the extent of oxygen deficient waters and nuisance algae and
declines in water clarity, submerged aquatic vegetation, finfish, and oysters (Flemer
et a/., 1983) were observed. Eutrophication was suspected to be the cause of many of
the problems. Though toxics were a major concern, there was much more uncertainty
about the cause-effect relationships (Magnien and Haire, 1989).
In 1983, the first Chesapeake Bay Agreement was signed by Maryland, Virginia,
Pennsylvania, the District of Columbia, and EPA. The agreement signified a commit-
ment to establish a structure to oversee cooperative and comprehensive measures
needed to restore the bay.
In 1984, the first basin-wide monitoring network was established along with a wide
range of new initiatives and legislation passed in all three States and the District of
Columbia. Previous monitoring information was insufficient to define the changes in
water quality between the 1950's and 1970's. Therefore a long term monitoring
program was designed to improve the understanding of the bay dynamics associated
with pollution reduction, to characterize the current status of water quality problems,
and to determine the long-term effects of management actions (e.g., phosphate deter-
gent ban, improved sewage treatment, nutrient reduction strategy, and best manage-
ment practices).
1987 Chesapeake Bay Agreement
The Chesapeake Bay Program has evolved over a number of years. The 1987 Chesa-
peake Bay Agreement was especially important because it set forth a water quality
goal to "reduce and control point and nonpoint sources of pollution to attain the water
quality condition necessary to support the living resources of the Bay." As a result of
this commitment by the Governors of Virginia, Maryland, and Pennsylvania, the
mayor of the District of Columbia, the Chesapeake Bay Commission, and the EPA
Administrator, bay-wide strategies for the control and reduction of nutrients, toxics,
and conventional pollutants were to be developed.
Nutrient Reduction Strategy
The 1987 Chesapeake Bay Agreement established a commitment to develop, adopt,
and begin implementation of a strategy to equitably reduce the total inputs of nitrogen
and phosphorus entering the mainstream of the Chesapeake Bay by at least 40% by the
year 2000. The target of 40% nutrient reduction was developed by modeling the
environmental conditions in the Chesapeake Bay with a two-dimensional, steady-state
model. According to modeling efforts, there are 1.35 billion cubic meters of anoxic
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A-27
water in the bay sometime during the summer and an average of less than 0.5 mg/l of
lowest summer dissolved oxygen in the deepest portions of the bay (based on 1985
nutrient loadings). Model results also suggest that a significant improvement in
dissolved oxygen in bay bottom waters can be achieved if nitrogen and phosphorus
loads are reduced. For example, the model predicts that after full implementation of
the 40% reduction goal that the lowest average dissolved oxygen will be 1.67 mg/t and
no waters will be anoxic.
Other benefits such as decreased algal biomass are expected as well. Chlorophyll
concentrations are expected to decrease (based on model predictions) from 13.6 u,g#
to 8.8 \ig/l. The reestablishment of rooted aquatic plants and improved fish popula-
tions are expected as well but cannot be quantified (Chesapeake Bay Program, 1988).
Model predictions are based on the information available in 1985 and the available
model did not include a mechanism to account for nutrient loading from sediment.
Since each state had a different mix of point and nonpoint source inputs as well as
different programs and policies, each state has a unique plan for nutrient reductions
(Chesapeake Bay Program, 1988). This approach allows each state to implement
individual strategies that are best suited for that state within the limits of its program
constraints, while still reaching the overall goal of 40% reduction at the bay-wide
level.
In addition, the reduction target is to be reevaluated by December, 1991 based on
modeling, research, and monitoring results.
Program Objectives
The first step in developing a bay-wide monitoring program was to review historical
data. Although there were some data shortcomings, the Chesapeake Bay Program was
nevertheless able to demonstrate degraded environmental conditions.
Three general program objectives were developed (Magnien and Haire, 1989):
• Quantify the extent and nature of water quality problems
• Determine the response of key water quality variables to management
actions
• Develop and test hypotheses on how the Bay ecosystem functions, espe-
cially as it relates to anthropogenic stresses and management solutions
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Based on eutrophication concerns and the complexity of the Chesapeake Bay, it was
also clear that the monitoring network would need to:
• Incorporate physical, chemical, and biological variables
• Collect mainstream and major tributary data
• Collect data consistently over an extended period of time
• Determine the optimal sampling strategy for each variable
As a result of data characterization and development of program objectives, six
monitoring program components were formed and are summarized below (Magnien
and Haire, 1989 and Chesapeake Bay Program, 1989a):
Chemical/Phvsical Variables: salinity, temperature, Secchi depth, dissolved
oxygen, suspended solids, nutrient species (nitrogen, phosphorus, carbon,
and silicon), phytoplankton pigments, heavy metal and organic compounds
in surficial sediments
Phytoplankton: species counts, phytoplankton pigments by horizontal and
vertical in vivo fluorescence, primary productivity, light penetration
Zooplankton: micro (44 urn - 202 um) and meso (>2Q2 |om) species counts,
biomass
genthic Organisms: species counts, production, sediment characteristics,
salinity, dissolved oxygen
* '
Ecosystem Processes: sediment-water column exchange rates of dissolved .
inorganic nutrients (nitrogen, phosphorus, and silicon) and oxygen,
surficial sediment characteristics, deposition rates of paniculate matter
(total seston, nitrogen, phosphorus, carbon, phytoplankton pigments)
River Tnputs: flow, suspended solids, nutrient species (nitrogen, phosphorus,
carbon, and silicon), phytoplankton pigments
Variable selection and spatial/temporal intensity were evaluated in order to provide the
level of information necessary to support confident management decision making. In
general, historical station locations were used as initial station locations and were
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"filled-in" with additional stations to provide adequate spatial coverage. Selected
sampling frequency was either monthly or bimonthly.
1991 Reevaluation Study
A review of the progress made toward the goals established in the Nutrient Reduction
Strategy will be completed by December 1991. Included in this review is the develop-
ment of a three dimensional dynamic model that will overcome previous limitations
and permit more precise estimates.
A2.3
Evaluation of
Monitoring
Program
Performance
The use of statistical power analysis to evaluate expected monitoring program perfor-
mance is demonstrated below using a subset of data from the Chesapeake Bay Moni-
toring program. The purpose of evaluating monitoring program performance is not to
determine whether the goals of the nutrient reduction program have been met but
rather to determine what is the minimum change that the monitoring program can be
expected to detect. This information can be used to address a series of questions
regarding the adequacy of the monitoring effort. Examples include:
• Are sufficient data being collected to detect a change in ambient nutrient
concentrations equal to the expected decrease?
• How long (e.g., number of years) will it take to detect the level of change
predicted by the numerical models?
This example includes a description of the data obtained from the Chesapeake Bay
Program. Several analytical techniques used to explore the statistical nature of the
data are also described. The purpose of these analyses is to identify characteristics of
the data that might affect the applicability of statistical tests. Next, statistical tests for
trends are described, and results of the analysis of the example data set are presented.
Statistical trend analyses were conducted in order to determine whether a trend could
be detected. Finally, a series of power analyses are then presented and discussed. It
should be noted that the results from the small subset of data used in this demonstra-
tion is not necessarily indicative of the expected performance of the Chesapeake Bay
monitoring program. An analysis of the nutrient data that has been collected from
throughout the bay would be required.
However, this example describes the use of general procedures can be applied to these
and other types of monitoring data.
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Example Data Set and Sampling Layout
For this example, data were obtained for dissolved oxygen and ammonium as nitrogen
from 15 stations. These stations were chosen because they provided adequate spatial
coverage as well as having similar sampling methodologies. The station locations
(four stations from segment 3, ten stations from segment 4, and one station from
segment 5) are shown in Figure A2-1. The principal characteristics of the segments
are (Flemer et al., 1983):
Segment 3
• Upstream limit of deep water anoxia
• Moderate salinity (7-13 ppt. mean)
• Two-layer, estuarine circulation driven primarily by freshwater inflow
Segment 4
• Water deeper than 30 feet, oxygen depletion in summer, potentially toxic
to fish, crabs, shellfish and benthic animals.
• Mean salinity of 9 to 14 ppL
• Rich in nutrients
Segment 5
• Influenced by inflow from the Potomac and Patuxent and rich in nutrients.
• Mean salinity of 10 to 17 ppt.
• Subject to summer anoxia and contains most of the deeper Bay waters.
Observations of the water quality random variables are collected bimonthly (monthly
during winter months), for a total of 20 samples per year at four station depths. These
depths are denoted as surface, above the pycnocline, below the pycnocline, and
bottom. When the estuary was not stratified at selected station locations, the
pycnocline data were collected at equally spaced distances between the surface and
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A-31
Figure A2-1. Station
locations included in data
subset of the Chesapeake
Bay Ambient Monitoring
Program.
Washington, DC
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A-32
bottom. Approximately 66 months (June, 1984 through November, 1989) of data were
available for each water quality random variable.
The subgroup of stations from the Chesapeake Bay Program was selected based on the
physical grouping provided by Hemer et al. (1983). In this example analysis depth is
used to group station locations. Further studies could be developed to separately
analyze each segment as defined by Flemer et al. (1983).
Exploratory Analyses
Before applying a statistical test for trend, several aspects of the time series need to be
explored. These include: distributional characteristics (i.e., deviations from normal-
ity), the existence of trends, seasonality, and serial correlation.
The choice of the appropriate statistical test will depend on which of these characteris-
tics the data exhibit The distributions of the data were shown to be non-normal. Time
series plots were examined visually for trend and seasonality. A series of monthly box
plots for each monitoring variable was made to examine the distribution of monthly
data and the seasonal pattern. Each box plot represents that month's data over six
years for a group of stations and shows the mean, median, and several percentiles (see
Figure A2-2).
Figures A2-3 and A2-4 show the time series plot for ammonia as nitrogen for the a
surface station and dissolved oxygen at a bottom station. The recurring sinusoidal
pattern indicates seasonality, a common occurrence in water quality data (Harcum
etal., 1990).
Figures A2-5 and A2-6 show the box plots for nitrogen as ammonia and dissolved
oxygen. If there was no seasonality, the means for all the months would be approxi-
mately the same. However, the monthly mean values vary depending on the month
which indicates that there is seasonal pattern.
The data were also tested for serial independence, (i.e., that the value at one timepoint
is dependent on the value of the previous timepoint). Most stations were found to
exhibit independence.
Statistical Tests for Trend
The choice of statistical test should be selected based on the data characteristics
described above. (Overall, the data were not significantly correlated; however, the
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A-33
data were seasonal and nonnormally distributed.) The Seasonal Kendall tau test was
selected since the test accounts for seasonality and is applicable to data that are not
distributed normally. Numerous researchers (e.g., Hirsch et al., 1982; Hirsch and
Slack, 1984; Berryman et al., 1988; Ward et al., 1988; Camoncho and Haywood,
1988; Taylor and Loftis, 1989; and Harcum et al., 1990) have also suggested that the
Seasonal Kendall tau test is an appropriate test for detecting gradual trends with the
type of data collected in the Chesapeake Bay Program and is typically more robust
over a wider range of conditions than alternative tests.
The Seasonal Kendall tau is a nonparametric test based on ranks. Each data point is
assigned rank based on its magnitude (smallest values get the lowest ranks, etc.). The
ranks are compared within a season (in this case, a season is a month) across the
years (May, 1984, May 1985, etc.) to determine if there is a trend. The results from
each season are then tallied and compared to a criterion value obtained from the
normal table.
The uneven sampling frequency of the example data set presents some potential
problems for the analysis described. Each sampling period (season) cannot be treated
equally since more samples were collected in the summer months. Additionally, the
Figure A2-2.
Example box plot.
Box Plots for Column X^
280 -
•a 260 -
8. 240 -
O 220 -
2 200 -
|* 180 -
'c
8 160 -
« 140 -
120 -
iftrt —
^ > median +
o 3 x interquartile range
o
o
° _ median +
1.5 x interquartile range
+ -mean
. interquartile
range
] ^ median -
~ 1 .5 x interquartile range
8
8
* ^ < median -
3 x interquartile range
-
Column
-------
A-34 g|
»B, f ? \ ^%v s ^^ •
•Mi * * tn fcrStf v>*>M 0 *s'
HT** s , v , J^
Jhh p w v*V ,. •. xv ,
0.12
O.OO
STATION CBS. 1
Apr-84 Jun-85 JuI-86 Aug-87 Sep-88 Oct-89
SAMPLE DATE
Figure A2-3. Time series
for N as NH3 for surface
stations.
x.
o
Q
u.
BOTTOM STATIONS
STATION ID
CB3.3E
o CB5.1
r-84
Jun-85
JuI-86 Aug-87
SAMPLE DATE
Sep-88
Oct-89
Figure A2-4.
Time series for Dissolved
O2 for bottom stations.
-------
^jSVjt'W
A-35
Figure A2-5. Box plot for
N as NH4 for surface
stations.
Variable
0.45-
0.40-
0.35-
0.30 -
0.25-
0.20 -
0.15-
0.10 -
0.05-
0.00-
= NH4(mg/L): Surface Waters
*
*
4
4
;
t
» * 0
n *
0
F
I
JAN
0 n
» (
0
>
+«H
. »..
^
)
I
-* T"
--* 4i..
i
) C
1
0 i
t
h I 1
••* 1
— i
^
* <
i
8
0
I
| * * | ,,. | *.-i
1
1 1 1
FEB MAR APR
) (
i
i
'
)
1
;
•t—
, !
|— * * * |
0
n
0 0
}
rj r
4 !::
)
i
7 i"
*. i *--
••* t—
3
—
*
(
T"
r
«**
1
— *•
. |
4
MAY JUN JUL AUG SEP OCT NOV
...
— t
1
:.....#
'
DEC
Figure A2-6.
Box plot for dissolved O2
for bottom stations.
Variable = DO(mg/L): Bottom Waters
18-
16-
14-
12-
10-
8-
6-
4-
2-
0-
*
0 (
aO I
1
»•-*"* 1 1 » — »
, 1. » — _, *.-(..*
* *
I r:
0 1
0 0 |
«•«-
*
*
!
1
"r
0
0
1 1 1
JAN FEB MAR APR
i
i
K-J
i
i i
• n n
*•-
— * *--
+--
--*
~+
MAY JUN JUL
i
...
t—
K«
i
«
K
«,
-•
'
;
h
AUG
h—
...
-H-
1 <
d
•«+
SEP
t—
t—
«-
4—
, U t
d U
_i
!
«-*
4
OCT NOV DEC
-------
A-36
likelihood of violating the independence assumption of statistical tests for trends is
increased since summer samples were collected more frequently.
Two typical solutions for handling the unequal sampling are 1) averaging or 2) sub-
sampling (e.g., average monthly values to one monthly value or discard one value per
month during summer). In this case, past research (Harcum et al., 1990) has shown
that averaging data is the preferred method for this type of a monitoring program. For
the analyses presented below, average values were computed for 12 seasons (i.e., one
value was computed for each month). Equal weight was given for each month for long
term trend detection.
For more information about the Seasonal Kendall tau test see Hirsch, et al. (1982),
Hirsch and Slack.(1984), or Taylor and Loftis (1989).
The results of the Seasonal Kendall tau tests are summarized in Table A2-1. At the
0.10 significance level, several stations showed statistically significant negative trends
in ammonium concentrations for both the surface and bottom samples. Positive trends
were also detected in dissolved oxygen concentrations in the bottoms waters at two of
the fifteen stations analyzed.
Several of the stations shown in Table A2-1 did not demonstrate a statistically signifi-
cant trend in ammonium or dissolved oxygen concentrations. However, the inability
to reject the null hypothesis (i.e., Ho: no trend in measured values) does not justify its
acceptance. The level of sampling effort or period of observation may not have been
sufficient to detect real trends in the data.
The reader is cautioned that the data preprocessing procedure used here is not appro-
priate for all applications. Consider a monitoring program composed of monthly
observations for the first five years and quarterly observations for the next five years.
It is not appropriate to average the monthly observations to quarterly values for
comparison with the last half of the data record. Rather, to apply the Seasonal Kendall
tau test, one must subsample from the earlier record (discarding two observations per
quarter) to create a uniform record. Depending on the exact circumstances, alternative
statistical tests may be more appropriate.
Statistical Power Analysis
Statistical power analyses were used to determine the minimum trend that could be
detected and therefore to determine the adequacy of the monitoring design.
-------
A-37
TABLE A2WL SEASONAL KENDALI, TAU JUBSIJC.T (Gfc&lO)
CB33E%
CB44C
CB4l%
CB4,1C
CB4<3C
CB4JE
CB43W
CB4.4
CB33E
CB44C
CB4.1E
CB44.W
CB42W
CB44E
CB4^W
CB44
Surface
Surface
Surface
Surface
Surface
Surface
Surface
Surface
No trend
Negative trend
Jsfo trend
No trend
ISfbtreBd
Ko trend
Negative trend
Jfettead
No trend
^^2E
CB4.2W
CB43C -
C3&4.3E
CB43W
CB4.4
C9B3>2
* CB3^E
3E
,C844, ,
Surface
Surface
Siafaee
Sorface"
Surface
Surface
Surface
Surface
Swtface:
Stafsee
Bottom
Bottom
Bottom
Bottom
Bottom
Bottom
Bottom
Bottom
BottWH
Bottom
Bottom
Bottom
No trend
No trend
Nofcead
No (rend
No tread
No trend
No scend ^
No trend
Noaead
NotrSid
No trend
No trend
No&read
No trend
No tread
No trend
No trend.
No trend
For some statistical tests, it is possible to develop analytical solutions to calculate
power based on the statistical characteristics estimated from data (e.g., U.S. EPA,
1987). However for nonparametric tests such as the Seasonal Kendall tau, the stan-
dard procedure for computing the test's power is to use Monte Carlo methods.
Monte Carlo simulation methods were used to evaluate the ability of the ongoing
monitoring efforts in Chesapeake Bay to detect specified levels of trend in water
quality variables.
The analyses were conducted in the following manner:
-------
A-38
1) Sampling spaces were defined to exhibit a specified magnitude of trend.
The distributional characteristics (e.g., mean and variance) of the sampling
population were otherwise similar to the monitoring data obtained from
Chesapeake Bay.
2) Random samples from the specified sampling space were generated by
computer programs. The simulated sampling frequency was the same
utilized in the Chesapeake Bay Monitoring Program. Sampling was
simulated for periods of 5 and 10 years.
3) Each sampling experiment was then repeated 500 times.
4) The simulated data in each experiment were analyzed using the Seasonal
Kendall Test, and the results of all 500 simulations were summarized to
obtain an estimate of the distribution of the test statistic. These results
were used to determine the percentage of simulations in which the null
hypothesis (no trend) was correctly rejected (i.e, the power of the test).
5) This series of steps was repeated for different magnitudes of trend, and the
results were summarized to obtain the plots of the probability of detection
versus magnitude of trend shown in Figures A2-7 and A2-8.
In Figures A2-7 and A2-8, four power curves are displayed for ammonia as nitrogen
and dissolved oxygen. For each pair of lines (the two solid or the two dashed lines),
the line to the right is the projected power for the station with the most variability of
the 15 stations and thus least powerful and the line to the left is the station with the
least variability and thus the most powerful. The results from the other 13 stations
would fall in between these two lines. The solid and dashed lines represent projections
for 10 and 20 years of data. As one would expect a significant improvement in trend
detection results from ten extra years of data.
Once the power relationships have been developed, selected results may be read from
the appropriate figure. From Figure A2-8, for example, dissolved oxygen trends in
surface waters on the order of 0.062 to 0.129 mg/^-yr can be detected with 10 years of
data at an 80% probability (using the Seasonal Kendall tau test with a = 0.10). With
20 years of data and the same group of stations, trends on the order of 0.021 to
0.042 mg#-yr can be detected with an 80% probability.
The results of these and similar analyses may now be used as a basis to modify, if
necessary, the monitoring program. Since this study evaluated a limited selection of
-------
A-39
Figure A2-7. Power
analysis for NasNH3 for
surface stations.
DC
UJ
O
Q.
Variable: Ammonium as N
Location: Surface
10 years of data
20 years of data
0.000
0.001 0.002
SLOPE (mg//-yr)
0.003
0.004
Figure A2-8. Power
analysis for dissolved O2
for bottom stations.
tc
UJ
I
Q.
minimum
variance
maximum I
variance
I Variable: Dissolved Oxygen
Location: Bottom
10 years of data
20 years of data
| — — minimum detectable
trend for a power
| level of 0.8
0.0
0.00
0.04
0.08
0.12
0.20
SLOPE (mg/Ayr)
-------
A-40
water quality random variables, it is not appropriate to make recommendations for the
Chesapeake Bay Program. An extended analysis of other water quality random
variables would be needed and balanced against other monitoring objectives, not just
long term trend detection.
The type of results presented in Section A2.3 can be used to evaluate the performance A2.4
of individual monitoring program components. By applying this type of analysis to all Use Of
program components, the management team can then begin to assess the strengths and Power Analysis
weaknesses of the overall monitoring effort. This information can be useful in deter- Results
mining where limited financial resources should best be applied.
With the type of information generated in the previous section or similar "what-if'
power analysis scenarios, some general questions can be answered:
• Which stations are most effective for detecting trends?
• Which water quality variables are the most useful for detecting trends?
By answering these questions, it is possible to modify the sampling frequency (to
collect more or less data), deleting or moving stations, or narrowing or expanding the
water quality random variables sampled. By coordinating decisions with the other
monitoring objectives, improved evaluation of the physical, chemical, and biological
integrity of the estuary is possible.
Batiuk, Rich. 1990. Chesapeake Bay Liaison Office. Personal communication.
Berryman, D., B. Bobee, D. Cluis, and J. Haemmerli. 1988. Nonparametric tests for
trend detection in water quality time series. Water Resources Bulletin, 24(3):545-556.
Box, G.EJP. and G.M. Jenkins. 1970. Time Series Analysis, Forecasting and Control.
Holden-Day, San Francisco, 553 p.
Brockwell, P.J. and R.A. Davis. 1987. Time Series, Theory and Methods. Springer-
Vcrlag, New York, 519 p.
A2.5
References
-------
A-41
Camacho, R. and H.C. Haywood. 1988. An Application of Power Analysis for
detection of trends in water quality. Interstate Commission on the Potomac River
Basin.
Chesapeake Bay Program. 1988. Basinwide Nutrient Reduction Strategy. Prepared in
Response to the 1987 Chesapeake Bay Agreement.
Chesapeake Bay Program. 1989A. Chesapeake Bay Basin Monitoring Program Atlas,
Volume I & H.
Chesapeake Bay Program. July, 1989. Water Quality Data Management Plan, Revi-
sion 1. United States Environmental Protection Agency, CBP/TRS 31/89.
Chesapeake Bay Program Monitoring Subcommittee. 1989. The State of the Chesa-
peake Bay, Third Biennial Monitoring Report -1989. 33 pages.
Elster, H.J. and W. Ohle. 1985. An Experimental Test of the Relative Significance of
Food Quality and Past Feeding History to Limitation of Egg Production of the Estua-
rine Copepod Arcartia tonsa. Arch. Hydrobiol. Beih. Ergebn. Limmol. 21:235-235.
Flemer, D.A., L. C. Davidson, K. Price, G. B. Mackiernan, and B. Johnson. 1983. A
monitoring and research strategy to meet management objectives. Appendix F, A
Framework for Action. Philadelphia: U.S. EPA.
Fulton, R.S. and H.W. Paerl. 1987. Toxic and Inhibitory Effects of the Blue-Green
Alga Microcystis aeruginosa on Herbiverous Zooplankton. J. Plankton Res.
9(5):837-855.
Gearing, J.N., P.J. Gearing, D.T. Rudnick, A.G. Requejo, and M.J. Hutchins. 1984.
Isotopic Variability of Organic Carbon in a Phytoplankton-Based, Temperate Estuary.
Geochim. Cosmochim. Acta. 48(5):1089-1098.
Grassle, J.P. and J.F. Grassle. 1984. The Utility of Studying the Effects of Pollutants
on Single Species Population in Benthos of Mesocosms and Coastal Ecosystems,
pp. 621-6432. In Concepts in Marine Pollution Measurement (HH. White, ed).
Maryland Sea Grant Publication, University of Maryland, College Park, MD.
-------
A-42
Grassle, J.F., J.P. Grassle, L.S. Brown-Leger, R.F. Petrecca, and N.J. Copley. 1985.
Sublidal Macrobenthos of Narragansett Bay, Reid and Mesocosm Studies of the
Effects of Eutrophication and Organic Input on Benthic Populations, pp. 421-434. In
Marine Biology of Polar Regions and Effects of Stress on Marine Organisms (J.S.
Grey and M.E. Christiansen, eds.). John Wiley and Sons, New York, NY.
Harcum, J. B., J.C. Loftis, and R.C. Ward. 1990. Selecting trend tests for water
quality series with serial correlation and missing values. Water Resources Bulletin, in
review.
Hirsch,RJM,J.R. Slack, and R.A. Smith. 1982. Techniques of trend analysis for
monthly water quality data. Water Resources Research, 18(1): 107-121.
Hirsch, RJM. and J.R. Slack. 1984. A nonparametric trend test for seasonal data with
serial dependence. Water Resources Research, 20(6):727-732.
Magnien,R.E. and M.S. Haire. 1989. Maryland's Chesapeake Bay Water Quality
Monitoring Program: An Estuarine Water Quality Information Systems. In Proceed-
ings of the International SYmposium on the Design of Water Quality Information
Systems. June 7-9, Colorado State University, Fort Collins, Colorado.
National Research Council. 1988. Marine Environmental Monitoring in Chesapeake
Bay. Marine Board, NRC, 2101 Constitution Avenue, Washington, D.C., 20418,
81 pages.
National Research Council. 1990. Managing Troubled Waters, The Role of Marine
Environmental Monitoring. National Academy Press. Washington, D.C., 125 pages.
Nixon, S.W., C.A. Oviatt, J. Frithsen, and B. Sullivan. 1986b. Nutrients and the
Productivity of Estuarine and Coastal Marine Ecosystems. J. Limmol. Soc. South Afr.
12(l/2):43-71.
Salas, J. D., J.W. DeUeur, V. Yevjevich, and W.L. Lane. 1980. Applied Modeling of
Hydrologic Time Series. Water Resources Publications, Fort Collins, Colorado, 484 p.
Taylor, C.H. and J.C. Loftis. 1989. Testing for trend in lake and ground water quality
time series. Water Resources Bulletin 25(4):715-726.
-------
A-43
U.S. EPA. 1987. Bioaccumulation Monitoring Guidance: Strategies for Sample
Replication and Compositing, Volume 5. OWXQMEP EPA 430/09-87-003. Washing-
ton, D.C.
U.S. EPA. 1990. EPA nonpoint Source News-Notes. #5. June, 1990. Washington,
D.C.
Verity, P.O. 1987. Factors Driving Changes in the Pelagic Trophic Structure of
Estuaries, with Implications for Chesapeake Bay, pp. 35-56. In Perspectives on the
Chesapeake Bay: Recent Advances in Estuarine Sciences (MP. Lynch and E.G.
Krome, eds.). Chesapeake Bay Program and Chesapeake Research Consortium, U.S.
Environmental Protection Agency, Gloucester Point, VA. CBP/TRS 16/87. CRC
Publication No. 127.
Ward, R.C., J.C. Loftis, H.P. DeLong, and H.F. Bell. 1988. Groundwater quality: a
data analysis protocol. Journal Water Pollution Control Federation, 60(11):1938-1945.
-------
A-44
-------
-------
-------
-X .v. • • f - v.ss ^V.
•.-.•... ^X s •. *•• %
•t ' - ^ • ^ - ^ ..\. - "^^ " ' v ' A««* *s\
Appendix B - Methods
-------
-------
B-3
.
*•¥.,•...
ViVMJ* WA
Methods - Introduction
This section identifies and describes sampling and analytical methods that are used in
estuarine monitoring programs. The primary purpose of this section is to provide
program coordinators and scientific staff of the estuary programs with a basic under-
standing of the sampling methods as well as information on feasibility and the use of
the monitoring program results. As stated in Section 4.1, standardized sampling and
analytical protocols or a performance based quality assurance program should be
developed for each estuary. This strategy is extremely important to ensure compara-
bility of data. The methods and recommendations described in this Appendix can be
used to guide decisions regarding the selection of individual techniques as well as
appropriate quality assurance and quality control procedures.
The primary sources of information for these method descriptions were documents
developed by the EPA's Office of Marine and Estuarine Protection and the Puget
Sound Estuary Program Protocols (described below). Individuals from EPA's Office
of Research and Development, the National Oceanographic and Atmospheric Admin-
istration, the U.S. Fish and Wildlife Service, university research laboratories and
ongoing National Estuary Programs were also contacted for information on latest
developments and ongoing research.
B.1 The methods that are described in this section (Table B-l) are grouped for presentation
Methods Chapter into four categories: water quality, sediment quality, biological resources and human
Format health risks.
The format that is used to describe the selected sampling and analytical methods is shown
in Table B-2. Following a brief description of the monitoring method and a definition
of terms, the rationale for monitoring is described in terms of what information will be
generated and how that information can be used to address the goals and evaluate the
success of the CCMP. Major monitoring design issues such as selection of species for
monitoring or selection of temporal and spatial sampling strategies are addressed under
the topic of monitoring design considerations. Selected existing laboratory QA/QC
and analyses methods are described. Statistical design considerations include method-
specific statistical techniques to evaluate expected monitoring program data. The data
use section provides guidance for data interpretation and use in addressing issues of
concern. All information presented for each monitoring program component is
summarized. Each chapter emphasizes the importance of coordinating monitoring
program components to assure an effective, cost-efficient monitoring program.
As part of the task of summarizing available information, emphasis is placed on
providing references to existing documents. Currently, there is a major cooperative
-------
B-4
1 % *, wSs^-> f s
^ Sj. K81 •* **f ^ s -^
"jWsrfw»YvXwj3sl5xvww^ •.-.h-. -. -I
TABLE B-l. SAMPLING METHODS DESCRIBED
Water Quality ;
2. Water Column Chemistry
x "*
Sediment Quality x
3. Sediment Grain Size
4* Sediment'CJtiemistry
Biological Eesources
5. Plankton
6. Aquatic Vegetation ,s "
7t BenthicTnfaunaCpnimunity Structure
8. Fish Community Stracmre
9, Fish and Shellfish Pathobiology
Human Health Risks _ - "••
10. Bioaccuniulation
11. Bacterial arid Viral Pathogens
effort underway within EPA, the U.S. Army Corps of Engineers, the National Oceanic
and Atmospheric Administration, and the National Bureau of Standards to develop a
Compendium of Methods forEstuarine and Marine Environmental Studies. The draft
document describes methods for nutrient measurements in seawater (U.S. EPA,
1990b). Additional sections are under development. Other important sources of
monitoring program guidance include documents developed by the Office of Marine
and Estuarine Protection for implementation of the 301(h) permit program. These
documents (Table B-3) provide useful information on sampling equipment, field and
laboratory methods, statistical techniques, and the design and implementation of
marine and estuarine monitoring programs. The Puget Sound Estuary Program, in a 2-
year effort, developed standardized protocols for measuring selected environmental
variables in Puget Sound (U.S. EPA, 1986-1991). These protocols have been devel-
oped for the monitoring topics shown in Table B-4. The process of developing these
regionally standardized protocols is described by Becker and Armstrong (1988).
-------
B-5
This methods section is also intended to provide a summary of available information
and to address the most important issues associated with the design and implementa-
tion of the monitoring program. Issues common to all monitoring methods include
quality assurance/quality control (QA/QC) and statistical design. These two important
issues are discussed below.
TABLE B-2. FORMAT OF METHODS SECTION :
Introduction _ "
' • description of method
• definition of terms •.•7- ,7
Rationale
~ • tyr^s of information generated
» applicability of ^formation
Monitoring Design Considerations
, * monitoring design issues
» guidance for the Implementation ofX ^ t
appropiate monitoring design; strategies 7 .. ss
Existing Analytical Methods
" • ixief overview of available laboratory and
data analysis "" - .. ..,
• ' advantages/limitations of methodologies
QA/QC Considerations
» selected QA/QC protocols % -
Statistical Design Considerations
• statistical: design strategies ~- ^ s^sw.
» guidance for the implementoBon of appropriate
statistical design
DataTOse - ," ---- -
« use / interpretation Of monitoring results
, * presentation of data
integration
Summary and Recommendations
Literature Cited and References
-------
B-6
s-V^*1 * „ v
TABLE B-3. TECHNICAL SUPPORT AND
GUIDANCE DOCUMENTS
Monitoring Program Development,
Implementation and Evaluation
Design of 30t(h) Monitoring Programs for Municipal"
Wastewater Discharges to Marine Waters, 1982,
'" % ~
^ " s
Ecological impacts of Jewage Discharges on Coral Reef
Communities. ' ^83^^ 430/^-8-010,
""•Xs1- ^ ^*>V^S V """V^s S ^
Initial Mixing Characteristics OfMunicipal Ocean
Discharges. 1985, EPA""" "s ~~ """
Summary of U.S. EPA-Appn>ved Methods, Standard Methods, and
Other Guidance for 3Ql(h) Monitoring Variables. 1985*
EPA 503/4-90-0021 ^ t S\,VV\^ ,„'!""? ' "*\ ,
Analytical Methods fort£S>EPA Priority Pollutants and 301(h)
PesdcidesinEstuarineandMarine^Sediments, 1986,
EPA503/6-90-004. \is^^ ' , ,.,
Evaluation of Survey^PpsjIoniJig Mfettioldf^ for Nearshom Marine and
Es^arine Watek'l987^E^X5430/9-86-ob3, ' ' ' """"('
'. ""•'W s
S s """ S **
Framework for 301 (h>Mbnitorihg Programs, 198>» -'-'" -
EPA430/09-88-002s, ,, - ->- '-^ - * ^^
Guidance for Conducting Fish Liver Histopathology StuBies during
301(h) Monitoring., 1987, EPA?430/9-87-004. ' 'VT" _
Quality Assurance and^Qualit^ Contrpf (QA/QC) for 301 (h)
Monitoring Programs: Outdance on Field and Laboratory'Methods,
1987,EPA43Q/9-86-004:':f % »-x . <:'""„,„
s ,_
••^s "^ s ^ ""• ** S "" "• "" -.
Recommended Biological Indices for 301(h) Monitoring Programs,
1987,EPA430/946-002; — /,. - ^ ->.,»>
- ' " •> Xs ^-. s %
Guide for Preparation of Quality Assurance Project Plans for the ''
National Estiiarine Program. Interim Final. 1988, EPA556/J-8>-06l,
-------
•KWVWAS TTTTf IVL t » » " jy*
r^^ N.^* ^ ^m. •*,
< ,' •• -4\\ ^ ™ .
B-7
TABLE B-3. TECHNICAL SUPPORT AND
GUIDANCE DOCUMENTS
(continued)
« A Simplified Deposition Calculation (DECAL) for Organic
Accumulation Near MaktaetfotBflfe. 19S7, EPA 430/09-88-0(51.
• Technical Support Document for ODES Statistical Power
Analysts. 1987, EPA 43G/9-87-Q05.
« Evaluation of Differential Loran-C for Positioning in
Nearshore Marine and EslSarine Waters. 1988,
TetraTech., Inc., Draft Report, EPA Contract No. 68-C8-OOi.
Bioaceumulation Monitoring Guidance Series
» Bioaccumulation Monitoring Guidance; 1) Estimating the
Potential forBioaccuinulation of Priority Pollutants and
3Ql(h) Pesticides Into Marine and Estuarine Waters,
1985, EPA 5Q3/3-9Q-OQL
* Bioaccumufatjon Monitoring Ouidance; 2) Selection Of
Target Species and Review of Available Bioacdumulation ^
Data. 1985, EPA 430/9-86-005, ' "
« Bioaccumulation Monitoring duidance: 3X Recommended
Analytical Detection Limits, 1985, EPA 503/6-90-003r '
» Bioaccumulation Monitoring <3uidancef4) Analytical
. Methods forXJ.Sx EPA Priority Pollutants and 301^i) Pes^cides-
In Tissue from Estuarine and Marine Organisms.
1986, EPA 503/3-90-001
^
*•"• ""• %%'.',-'
« Bioaccumulation Monitoring Guidance; 5) Strategies fot
Sample Replication andCompositing,
1987, EPA 430/9-87-003,
-------
B-8
TABLE B-l.
MONITORING TOPICS INCLUDED IN PUGET SOUND
ESTUARY PROGRAM PROTOCOLS
General QA/QC Considerations for Collecting Environmental Samples
Measuring Conventional Sediment Variables,
Conducting Laboratory Bioassays \, ' ,
SSN % , ,""
Station Positioning % -,
5 ' •"'"•••»
Measuring Metals in Water* Sediment, and Tissue Samples"
Microbiological Studies > - «• ^
s • j.j '"^ •,
Measuring Organic Compounds in Sediment and Tissue Samples
Sampling and Analyzing Subtidal Benthie Macroinvertebrate Assemblages
Fish Pathology Studies " - ? « ^^ ••
Measuring Conventional Water Quality Variables and Metals in Fresh Waters
Sampling Soft-Bottom Demersal Fishes by Beach Seine and Trawl
Measuring Conventional Marine^Water Column Variables.,
v -,\ •> -f
Marine Mammal Tissue Sampling and Analysis ,^wm
The purpose of Quality Assurance is to ensure that the environmental monitoring
efforts result in data of known and acceptable quality. There are two types of quality
assurance plans: the Quality Assurance Program Plan (QAPP) and the Quality Assur-
ance Project Plan (QAPjP).
Quality Assurance Program Plan. Development of a QAPP is important in estab-
lishing the overall framework for implementation of quality assurance within a
specific estuary program. It establishes the quality assurance policy, overall proce-
dures and responsibilities.
B.2
Quality
Assurance/Quality
Control (QA/QC)
Considerations
-------
B-9
*t f •• %K
S\X;., f ••xftv^v.sv.
Aft1- J»i
-9''
Quality Assurance Project Plan. Quality Assurance Project Plans must be prepared
for all projects involving environmentally-related data as a requirement of both EPA
and National Estuary Program policies. TheQAPjPis a recognized tool for providing
the detailed information needed to ensure the reliability and comparability of data
collected during a project. Guidance on developing a QAPjP is provided in U.S. EPA
(1988c). Examples of some key considerations are provided in Table B-5.
Data Quality Objectives
A major part of the QA effort is establishment of data quality objectives (DQOs).
DQOs are specific, integrated statements and goals developed for each data or infor-
mation collection activity to ensure that the data are of the required quantity and
quality. DQOs should specify the desired sensitivity of sampling methods, timing and
TABLE B-5,
KEY TOPICS TO BE ADDRESSED
IN ESTUARY PROGRAM QUALITY ASSURANCE PLANS
» Instructions for proper sample preservation and storage
Chain of custody documentation
» Source of standard reagents " -
* Source orpreparation of quality control samples "
» Frequency of analysis of quality control samples
* v Data Quality Objectives
» Analysis of QC samples
» Review of results
* Corrective action when unsatisfactory results are obtained in the analysis of
,, QA/QCsamples ~ """ "\\ _,,,,,.
Calihrattort and maintenance of equipment
-------
B-10
location of sampling, and numbers of samples to be collected. One function of
establishing DQOs, as discussed in Section 2.3, is to ensure that monitoring/
data collection studies yield data adequate to meet their intended uses. DQOs
are also used to delineate QA/QC programs specifically geared to the data
collection activities to be undertaken. Development of DQOs usually consists
of three processes: 1) decision definition, 2) data use and needs identification,
and 3) data collection program design. DQOs should be continually reviewed
during data collection activities and any needed corrective action may be
planned and executed to minimize problems before they become significant.
DQOs play an important role in the selection of sample collection, sorting, and
analysis methods. In fact, the selection of monitoring methods should be driven
by DQOs. Any future modifications to monitoring protocols should be consid-
ered only after these new methods meet established data performance criteria.
Data collected with different methods should not be compared unless informa-
tion exists which supports such comparisons.
The following sections provide a discussion of existing monitoring methods.
These methods provide a starting point for considering monitoring methodolo-
gies. New, cost-effective methods are sure to develop for certain disciplines in
the coming years. A key consideration when deciding whether new methods
are to be incorporated into the monitoring program, is whether they meet the
QA/QC program's performance criteria. QA/QC considerations in the following
sections will primarily discuss sample collection, processing, storage, and
analysis.
QC Samples
Additional samples, either collected in the field or prepared from standard
reference materials are used to check for field or laboratory contamination, test
laboratory accuracy, test field or laboratory reproducibility, and assess natural
variability. There are four basic types of QC samples that should be submitted
with each set of sediment or water quality samples (Table B-6). Although the
frequency with which these samples are collected will vary among programs,
each should be collected at least once for every 20 samples or sampling event,
whichever is more frequent.
Data from existing monitoring programs will often be utilized in an estuary
program. Therefore, data will frequently be collected by a number of agencies
following different sampling and analytical strategies. Under such conditions
-------
B-11
TABLE B-&
QC SAMPLE TYPES
Travel (Trip) Blank assess cross-contaminatJott during handling and transport
assess contaialiiatiottdae to teco
-------
B-12
dation, equal numbers of samples should be collected whenever possible to simplify
statistical analyses.
Composite Sampling
Composite sampling consists of mixing two or more replicates and/or samples. The
chemical analysis of a composite sample provides an estimate of an average contami-
nant concentration for locations and/or times comprising the composite sample (see
space and time bulking below). Advantages of the composite sampling strategy are:
• provides cost-effective strategy when individual chemical analyses are
expensive
• results in a more efficient estimate of the mean at specified sampling
locations.
Because of the reduced sample variance, composite sampling may result in a consider-
able increase in statistical power. If the primary objective of a monitoring program is
to determine differences in contaminant concentrations among sampling locations,
composite sampling is an appropriate strategy.
Space-bulking consists of sampling from several locations and combining samples into
one or more composite samples. Time-bulking involves taking multiple samples over
time from a single location and compositing these samples. A disadvantage of using
space- and/or time-bulking strategies is that significant information concerning spatial
and temporal heterogeneity may be lost.
Compositing of tissues provides a means to analyze bioaccumulation when the tissue
mass of an individual is insufficient for the analytical protocol. If mixing species,
tissue composites are likely to be composed of different proportions of species, and
different numbers, age, and sex of individuals. These differences among composites
tend to confound whether patterns of tissue residue concentrations are due to differ-
ences in locations or interspecific differences in bioaccumulation. Mixing of species is
not recommended because of these confounding effects.
Composite sampling is not recommended if the objective of the monitoring program is
to determine compliance with specified sediment contaminant concentration limits,
since this sampling method does not detect the true range of sediment contaminant
-------
B-13
-X
\ %
concentrations in the environment The adoption of composite strategies will depend
upon the objectives of individual monitoring programs.
Statistical Power
The large degree of temporal and spatial variability observed for many ecosystem state
and rate variables requires collection of sufficient replicate samples to ensure an
accurate description of the measure of interest. However, increases in replication
increase sample processing costs. Power analyses assist in the allocation of sampling
resources (stations, replication, and frequency) with regard to program finances and
design (Sokal and Rohlf, 1981).
Power analyses may be applied to determine the appropriate number of sample repli-
cates, and/or subsamples in a replicate composite, required to detect a specified
difference (U.S. EPA, 1987b). The number of replications required to detect a speci-
fied minimum difference is a function of the statistical power and the variance in the
data. Power analyses require a prior knowledge of the variability in the data. A best
guess or variation observed in historical data are often used initially in the design of
the monitoring program.
For composite samples, statistical power increases with the increase of the number of
subsamples in each replicate composite sample (U.S. EPA, 1987c). However, there
exists a diminishing return of statistical power with the addition of successive
subsamples to each composite. For composites of greater than ten replications, the
increase of power is negligible given typical levels of data variability.
To improve the power of a statistical test, while keeping the significance level con-
stant, increase the sample size. However, due to constraints in cost and time imposed
by the CCMP this option may not be available. Power analyses have shown that for a
fixed level of sampling effort, a monitoring program's power is generally increased by
collecting more replicates at fewer locations. The number and distribution of sampling
locations required to evaluate the effectiveness of the CCMP will be depend upon the
size and complexity of the estuary.
Power-Cost Analyses
The relative power of one design with respect to another is more meaningful when the
relative costs of implementing alternative designs are taken into consideration. Analy-
ses of power-costs are fundamental in selecting appropriate sample/replicate number,
-------
B-14
sample location, and sampling frequency (Bros and Cowell, 1987; Cuff and Coleman,
1979; Ferraro et al, 1989; Millard and Lettenmaier, 1986).
A cost function which describes the cost per replicate sample is required. Power-cost
formulations for parametric statistical analyses are of the form:
Power-costj = (1 - Bj) / (cj x n;)
where i is a replicate sample collection and analysis scheme, c is the cost per replicate
sample, n is the number of replicate samples, and IJ| is the Type II error. Costs will
depend upon sample collection, sorting, and analysis equipment and protocols. Type
II error (B) is a function of the number of replicate samples and the variance detected
by a sampling and analysis scheme. The measured variance may be influenced by the
sampling equipment used and the unit replicate sample size (e.g., as the unit replicate
area increases, variance increases).
Total costs are usually fixed. Therefore, iterations of power-cost formulations for
various sample processing scenarios (e.g., sample collection, sorting, and analysis
schemes) and their comparisons will result in the selection of the appropriate monitor-
ing methodology. Sokal and Rohlf (1981) provide a series of formulas for calculating
the most efficient sampling design for a given cost.
APHA. 1989. American Public Health Association, American Water Works Associa- B.4
tion, and Water Pollution Control Federation, Standard methods for the examination of Literature Cited
water and wastewater, 17th ed. American Public Health Association, and References
Washington, D.C.
Becker, D.S. and J.W. Armstrong. 1988. Development of regionally standardized
protocols for marine environmental studies. Mar. Poll. Bull. 19(7):310-313.
Bergstrom, P. 1990..Chesapeake Bay Coordinated Split Sample Program Annual
Report, 1989. Chesapeake Bay Program, Annapolis, M.D.
Bros, W.E. and B.C. Cowell. 1987. A technique for optimizing sample size (replica-
tion). J. Expt. Mar. Biol. Ecol. 30:21-35.
Chesapeake Bay Program. 1991. Coordinated Split Sample Program Implementation
Guidelines, Revision 3. EPA Chesapeake Bay Program, Annapolis, M.D.
-------
B-15
•«* "•-*-- ,y;%*fr ' "\
f "'""' ^2. •••••• ^ ^st: .',,1 i>x
Cuff, W. and N. Coleman. 1979. Optimal survey design: Lessons from a stratified
random sample of macrobenthos. J. Fish. Res. Board Can. 36: 351-361.
Ferraro, S.P., F.A. Cole, W.A. DeBen, and R.C. Swartz. 1989. Power-cost efficiency
of eight macrobenthic sampling schemes in Puget Sound, Washington, USA. Can. J.
Fish. Aquat. Sci. 46: 2157-2165.
Gilbert, R.O. 1987. Statistical Methods for Environmental Pollution Monitoring.
New York, NY: Van Nostrand Reinhold Co.
Hirsch, R.M. 1988. Statistical methods and sampling design for estimating step trends
in surface-water quality. Water Res. Bull. 24(3): 493-503.
Millard, S.P., and D.P. Lettenmaier. 1986. Optimal design of biological sampling
programs using the analysis of variance. Est. Csfl. Shelf Sci. 22: 637-656.
Sokal, R.R., and F.J. Rohlf. 1981. Biometry. San Francisco, CA: W.H. Freeman
and Co. 859pp.
U.S. EPA. 1983. Methods for Chemical Analysis of Water and Wastes, 2nd Edition.
EPA 600/4-79-020. U.S. Environmental Protection Agency, Environmental Support
Laboratory, Cincinnati, OH.
U.S. EPA. 1986-1991. Recommended protocols for measuring selected environmen-
tal variables in Puget Sound. Looseleaf. U.S. Environmental Protection Agency,
Region 10, Puget Sound Estuary Program. Seattle, WA.
U.S. EPA. 1987a. Quality Assurance/quality control (QA/QC) for 301(h) monitoimg
programs: Guidance on field and laboratory methods. EPA 430/9-86-004. Office of
Marine and Estuarine Protection, Washington, D.C. 267 pp.
U.S. EPA. 1987b. Technical support document for ODES statistical power analysis.
EPA 430/9-87-005. Office of Marine and Estuarine Protection, Washington, D.C.
34pp.
U.S. EPA. 1987c. Bioaccumulation monitoring guidance: Strategies for sample
replication and compositing. Vol.5. EPA 430/9-87-003. Office of Marine and
Estuarine Protection, Washington, D.C. 51 pp.
-------
B-16
LV^WWWK*
U.S. EPA. 1988. Guide for Preparation of Quality Assurance Project Plans for the
National Estuary Program - Interim Final. EPA 556/2-88-001. U.S. Environmental
Protection Agency, Washington, D.C.
U.S. EPA. 1990a. U.S. EPA Statement of work for organics analysis: multi-media,
multi-concentration. Document no. OLM01.0. U.S. Environmental Protection Agency,
Contract Laboratory Program. Washington, D.C.
U.S. EPA. 1990b. Compendium of methods for marine and estuarine studies. EPA503/
2-89-001. U.S. Environmental Protection Agency, Office of Water. Washington, D.C.
U.S. EPA. 1991. Statement of work for inorganic analysis: multi-media, multi-concen-
tration. Document no. ILM02.0. U.S. Environmental Protection Agency, Contract
Laboratory Program. Washington, D.C.
Ward, R.C. and J.C. Loftis. 1986. Establishing statistical design criteria for water
quality monitoring systems, review and synthesis. Water Res. Bull. 22(5):759-767.
-------
B-17
\
\
B1.0 Water Column Physical
Characteristics
B1.1 Physical characteristics of the water column are typically monitored to evaluate
Rationale estuarine water quality. The objective of monitoring these physical characteristics is to
identify and detect spatial and temporal variations in water quality. These results may
be used to characterize the water column, to monitor estuarine hydrodynamics, and to
verify estuarine hydrodynamic models.
Many chemical and biological processes in the environment are affected, directly or
indirectly, by the physical characteristics of the environment (Thomann and Mueller,
1987). Collection of physical data also provide ancillary information necessary in
evaluating chemical and biological data.
Considerations
B1.2 Monitoring schemes for physical characteristics usually involve in situ methods. Data
Monitoring Design concerning the water column's physical characteristics can be collected at relatively
inexpensive costs; the expense of different methods is generally governed by the level
of automation. Typically, the location and frequency of sampling water column
physical characteristics are determined by other, more expensively collected water
column parameters. However, it is essential to standardize the monitoring design in
order to ensure the comparability of monitoring data throughout the program.
Sampling Depths
When using bottle sampling techniques, samples for temperature, salinity, and turbid-
ity should be taken at a minimum of four depths in the vertical profile: 1) one meter
below the surface, 2) one meter above the bottom, 3) one meter above the pycnocline,
and 4) one meter below the pycnocline. If the waters are too shallow or no stratifica-
tion occurs, it would be appropriate to take the latter two samples at evenly spaced
distances between the top and bottom samples. However, features of water masses
recorded in the historical data (i.e., historical profiles of salinity, temperature, and
turbidity) and the collection of other data types (e.g. plankton community structure and
water chemistry) should be considered when establishing sample depths (Pond and
Pickard, 1983).
When using in situ methods [e.g., Conductivity-Temperature-Depth (CTD)] tempera-
ture and salinity measurements should be taken at one meter intervals over the entire
depth profile (to within 1 meter of the surface and bottom). Little additional cost is
incurred for this detailed characterization of the water column once the CTD is de-
ployed. In areas of high stratification, a smaller interval would be appropriate.
-------
B-18
hill In ii 11'11 :
MR <*•!•«*.
A consistent vertical sampling design is recommended in order that comparisons may
be conducted between data collected at different locations and over time. For ex-
ample, a set of depths may be specified by the monitoring design; at a minimum, any
sampling of the water column must include data from this set of specified depths.
Spatial and Temporal Scale
Generally, as the scale and complexity (spatially and/or temporally) of circulation
increases, more meters need be deployed to adequately describe the environment. The
need formacroscale and detailed descriptions of circulation will depend upon the
objectives of the CCMP and the physical structure of the estuary. Circulation patterns
and tidal dynamics may result in the establishment of fronts (i.e., regions of steep
gradients) and the accumulation of pollutants and organisms (Dustan and Pinckey,
1989; Pinckey and Dustan, 1990).
The length of the sampling period will be determined by monitoring objectives. If
descriptions of seasonal variations and/or high frequency natural events (e.g., tides,
storms, etc.) are necessary, measurements should be collected over a period not less
than two years (Fischer et al., 1976). Frequent sampling in fish and shellfish areas,
and other biologically sensitive areas may be required in order to detect rapid changes
in those habitats which may endanger these resources.
Moored instrument arrays provide a cost-effective method of continual sampling over
long periods of time (from days to months) for many physical characteristics. More
commonly used at remote offshore locations, subsurface arrays can provide unattended
long-term data collection in estuarine environments for both physical and chemical
parameters. Water depth, velocity (speed and direction), temperature, and conductiv-
ity or salinity are the most commonly measured characteristics. Estimates of total
suspended solids and chlorophyll can be recorded also, using turbidometers, transmis-
someters or fluorometers. Dissolved oxygen concentrations can also be measured.
Sensors capable of in situ measurements of nutrient and trace contaminant concentra-
tions are under development.
Moored arrays have the potential to provide synoptic coverage at pre-determined
intervals, which can range from minutes to days. The longer the interval between data
recordings, the longer the array can function without replacing the power supply
(usually batteries) or the recording medium (cassette tape or computer memory chips).
Real-time telemetry of data to a shore station is possible with the more sophisticated
(and more expensive) instrumentation packages. More commonly, the instrument
array is retrieved at regular intervals and the collected data post-processed.
-------
B-19
Moored arrays that are deployed for long periods of time are susceptible to many types
of interference. Within estuaries, deployment must take into consideration the impacts
of strong tidal flows, vessel traffic, unintentional interference from sub-surface nets,
fishing lines and anchor lines, and from intentional interference, such as theft and
vandalism.
B1.3
Existing Analytical
Methods
Methods available for monitoring physical characteristics include instruments that
range from simple mercury-filled thermometers that provide one observation of
temperature to state-of-the art CTD (Conductivity-Temperature-Depth) equipment that
provide vertical profiles of salinity and temperature. Methods (and/or equipment)
discussed in this section are summarized in Table B1 -1.
Temperature
Surface temperature measurements may be made with any good grade of mercury-
filled centigrade thermometer, or a thermistor taking care not to expose the water
sample to the sun or to the evaporative cooling effect of the wind (APHA, 1989). For
measuring subsurface temperatures the basic instrument is the "protected reversing
thermometer"; a mercury-in-glass thermometer which is attached to the water sam-
pling bottle (Pond and Pickard, 1983).
Bathythermographs (BTs) consist of a thermistor and an electronic pressure trans-
ducer. Wires extending from the BT to the vessel relay measures of temperature and
pressure. BTs may be operated from stationary or slow moving (< 4 knots) vessels.
Expendable bathythermographs (XBTs) are based on tracking an expendable, free-
falling bathythermograph. The system is designed to operate from a stationary or
moving vessel (< 15-20 knots) typically in deep waters. Wires extending from the
free-falling transducer to the vessel relay temperature and depth recordings. The wires
are cut when the probe reaches its maximum depth. BTs and XBTs do not record
measures of salinity. If it has been determined that density is not related to salinity,
bathythermographs may be a cost-effective tool. Monitoring of temperature and
pressure using XBTs is higher in cost since the probe is lost after each vertical profile.
Conductivity-Temperature-Depth (CTDs) meters are the tool of choice, since density
measures are typically dependent on salinity, temperature, and depth (see Density
below). Measures of density are important in identifying and tracking the movements
of water bodies and in determining the depth at which a water mass will settle at
equilibrium. CTDs coupled with a computer can continuously integrate temperature
and conductivity measurements with depth. CTD/computer provides real-time read-
-------
B-20
TABLE Bl-1. LIST OF METHODS AND EQUIPMENT
Temperature ;
• mercury-filled centigrade tftfemomete*
• thennistor
• reverse thermometer
• bathythermograph
• CTD 's *™
Salinity
Density
Depth
Knudsen titration 0,
conductivity cells (CTD instruments) ^
direct (hydrometers, pycnometers, magnetic float densimeters)
indirect (function ofsalinityj temperature, and pressure)
• fathometer ,
• meter wheel
• hydrostatic pressure
- bourdon tube ; -
- strain gauge \
Turbidity " ••,
• light transmissometer
• SccchidiSc
* nephelometer
• turbidimeter
., s-> %
Current Measurements
• Eulerian method (speed,ahd direction)^
- mechanical
- electromagnetic s -
- acoustic "
• Lagrangian method
» drogues
- surface drifters '"
- seabed drifterslC ^
Dye Studies
• fluorometers
-------
B-21
outs which allow immediate modifications in CTD position and ensures comprehen-
sive water column coverage during the survey.
Salinity
The Knudsen titration method determines chlorinity by titration with a standard silver
nitrate solution and salinity is then computed from a standard formula (Pond and
Pickard, 1983). This titration method is practical but not convenient to use on board a
ship (APHA, 1989).
Conductivity cells are used for in situ use and a variety of such instruments are now
available from several manufacturers (Pond and Pickard, 1983). A CTD unit, consist-
ing of conductivity, temperature, and depth sensors, is lowered through the water on
the end of an electrical conductor cable which transmits the information to recording
units on board ship (UNESCO, 1988).
Density
Density is an important identifying property of sea water because there is no process,
except mixing, by which this property changes below the surface (Pickard and Emery,
1982). Direct measures of density are slow (Pond and Pickard, 1983). Devices such
as hydrometers, pycnometers, and magnetic float densimeters may be used when great
accuracy is not required.
In practice, the density - or the specific gravity - of sea water can be determined from
temperature, salinity, and pressure. Temperature, salinity, and pressure are measured
directly and the density is read from tables which have been prepared from laboratory
determinations (e.g., Eugene, 1966). Algorithms for automated data processing have
been developed by UNESCO and are applicable for salinities as low as 2 ppt (Fofonoff
and Millard, 1983). For lower salinities, see Hill et al. (1986).
Depth
The simplest method of determining the depth of an instrument is to pass the wire
attached to the instrument over a meter wheel with a known circumference and a
counter. In calm conditions and negligible currents, the length of wire passed over the
meter wheel will be the actual depth.
Depth may also be estimated using a bourdon tube moving the slider of an electrical
potentiometer or a strain-gauge pressure transducer. The hydrostatic pressure exerted
-------
B-22
**'
on gauge or bourdon tube may then be converted to a depth (Pickard and Emery,
1982).
The water depth at a particular station is typically determined with acoustics. Depth-
sounders have accuracies of 0.3 to 0.7 ft (Clausner et al., 1986).
Turbidity
To determine the transmission of visible light through the water, the simplest device is
the Secchi disc (Pond and Pickard, 1983). Measurements of Secchi disc depth are
probably the most widely used means of estimating light penetration. The Secchi disc
is easy to use and can be used to estimate the attenuation coefficients for collimated
and diffuse light As a result, the depth of the euphotic zone may be estimated. Secchi
disk readings vary with the observer because of interpersonal differences in visual
ability and, therefore, caution must be exercised when comparing Secchi disk readings
taken by different observers.
Turbidity is a measure of the absorption and scattering of light by paniculate matter
(plankton and suspended sediments) as it passes through water. Measurements may be
either in situ or remote. In remote measurements, water samples are obtained from
discrete depths and then analyzed on deck or in the laboratory with a device such as a
nephelometer. This method of sampling is quick, but is labor intensive and is re-
stricted to sampling only at those depths where water samples were taken.
In situ sampling involves passing a light transmissometer or turbidimeter (usually as
part of a CTD system) through the water column. This provides for much higher
sampling rates and less labor. To correlate data taken from different instruments all
values should be expressed consistently as a percent transmittance for a standard
pathlength.
Nephelometry is the preferred method for measuring turbidity. Because nephelometers
measure light that is refracted at 90° as it passes throght the sample, they are more
sensitive, and effective over a wider range of turbidities than other turbidimeters. The
90° detection angle is also less sensitive to variations in particle size. Turbidity mea-
sured using nephelometric methods is expressed nephelometric turbidity units (MTU).
Current Measurements
There are two basic ways to describe fluid flow, the Eulerian method in which the
velocity (i.e., speed and direction) is stated at every point in the fluid, and the Lagrangian
method in which the path followed by each fluid particle is stated as a function of time.
-------
i'-v-v.v.v.
B-23
Eulerian Methods - Mechanical meters include Savonious rotors, ducted impellers,
drag inclinometers, and propeller-type meters. Although Savonious rotors have been
used for many years in estuarine work, they are not suited for operation in shallow
water environments which are exposed to wave and swell activity. Ducted impeller
current meters have been designed to reduce the problems associated with current
measurements in the presence of waves (Brainard and Lukens, 1975). Davis-Wellar
propeller type meters are also designed to operate in the presence of waves.
Electromagnetic current meters measure the instantaneous horizontal and vertical
velocity components at a flow sensor. The characteristics of these meters make them
suitable for use in the presence of waves and shallow water (McCullough, 1977).
Most recent electromagnetic current meters are equipped with several probes to
measure other physical characteristics of the receiving water.
Acoustic current meters determine the current velocity by emitting a short acoustic
pulse and measuring the Doppler frequency shift of the backscattered signal reflected
from small, passively transported particles in the water column (Appell and Curtin,
1990; Appell and Woodward, 1986). This instrument allows for non-intrusive, remote
measures of current velocity throughout the water column from a depth of 3 to over a
1000 meters. The current meter can be placed on a ship and sampling conducted while
the ship is underway, thereby allowing for a synoptic measurement of the current field
over a large area with the use of only one instrument. Acoustic current meters are
suitable for use in the presence of waves but the accuracy of the measurements de-
creases close to the water surface. These instruments are also expensive and require
highly trained technicians to maintain them.
Another technology employs the use of land based radar systems sending out electro-
magnetic waves and measures the reflected energy from the surface waves (Appell and
Curtin, 1990; Appell and Woodward, 1986). Since this system is set up on the shore it
only works out to a few dozen kilometers offshore and only measures the current
velocity in the top meter of the water column. This system does have the advantage in
that a ship is not needed to deploy or retrieve the instrument.
Lagrangian Methods - Most drogues are drogue-buoy systems consisting of a small
marker buoy which is tracked at the surface and a larger submerged drogue portion
which is set at the desired depth by connecting line between the two portions (U.S.
EPA, 1982). The drogue portion must be weighted and ballasted so that the drogue
assembly has sufficient negative buoyancy to keep both the drogue and connecting line
in their intended vertical orientation, and keep the buoy mast upright. Drogues are
intended to passively drift with the currents at a specified depth. In reality, some error
is introduced in the drogue trajectories by wind drag on the exposed portion of the
-------
B-24
marker buoy, by the relative surface current drag on the submerged portion of the
surface buoy, and, for deep drogues with long lines, by the relative current drag on the
connecting line. If possible, it is best to avoid drogue studies under high wind condi-
tions, especially when measuring lower current speeds in deeper waters. Drogues can
be used from just a couple of days to many months and can be tracked by satellite.
Most drogues can be classified into one of the following four categories: 1) Parachute
drogues, 2) Cruciform drogues, 3) Window Shade drogues, and 4) Cylindrical drogues.
Of these types, parachute and cruciform drogues are the most widely used (U.S. EPA,
1982).
Surface drifters are used to measure the average path of currents at the surface. Drift
bottles and drift cards are the types most commonly used. Vertical drift bottles and
drift cards are useful for evaluating the movement of effluent which reaches the
surface. Horizontal drift cards, which float on the surface, may be used to determine
the potential movement of surface slicks due to oils or other floatables which form a
surface film. Because these movements are influenced largely by the wind, horizontal
drift cards should be released under several different conditions (Grace, 1978).
Seabed drifters measure the average path of currents near the sea floor. They are
useful for determining the fate of waste materials subject to transport by bottom
currents. This includes setfleable solids and any portion of the effluent which remains
near the bottom. The drifters provide information on the net movement of a waste
field along the bottom, including where the waste field may reach the shoreline, and a
rough estimate of how long it may take. If a sufficient number of drifters are recov-
ered, they may indicate areas of possible shoreline contamination (Grace, 1978).
All environmental monitoring programs should have a written and approved quality
assurance project plan (U.S. EPA, 1987a). Water column sampling devices should be
checked for proper functioning prior to the survey. It is prudent to have a backup
sampler onboard the survey vessel in case the primary sampler is found to be unsuit-
able during the cruise.
Tables Bl-2 and Bl-3 lists sample preservation and storage requirements and recom-
mended methods for salinity and temperature (U.S. EPA, 1987a). When several meth-
ods arc available, the selection should be made by comparing the accuracy and precision
of the candidate methods for the parameter range expected at the site. All monitoring
programs should follow manufacturers recommended calibration procedures.
B1.4
QA/QC
Considerations
-------
B-25
* "AM*" '
TABLE Bl-2. RECOMMENDED SAMPLE PRESERVATION,
AND STORAGE REQUIREMENTS
Volume
Parameter "Required (ml) Container
Steservative BbWingTjine
Salinity
240
Temperature 1,000
0(with
paraffined
corks
air tight)
3VS
None'
,
(Loagerf
properly Dialed)
Etetetmiae " Honolding
on site
References: Standard Methods (APBA* 1989) and Methods for Chemical Analysis of
Water and Wastes fU.S. EPA. 1983> - w %
Note: ,P ^Polyethylene, G = Glass
Each temperature-measuring instrument should be calibrated against a precision
thermometer certified by the National Bureau of Standards at least every week during
sampling (U.S. EPA, 1986-1991). It is recommended that calibration be conducted
more frequently when temperature variation is suspected to ensure that readings reflect
environmental differences, not instrument drift.. Temperature probes may, at best, be
accurate to within one-tenth of a degree Celsius. The required sensitivity will depend
on the amount of natural variability in the environment and the amount of difference
which needs to be detected. Temperature probe systems are rarely linear over large
temperature ranges and must be checked against research grade laboratory thermom-
eters (APHA, 1989).
Salinity probe systems offer moderate accuracy but should be cross-checked by
discrete water samples analyzed by induction type laboratory salinometers (APHA,
1989). Two standard determinations should be made before the start of each series of
samples. In addition, one standard sample should be analyzed, to monitor instrument
-------
B-26
$*J*&b*v*&iv*v^Mww£ivif.\'.\v,v&l''*. v
TABLE Bl-3.
RECOMMENDED ANALYTICAL METHODS
Parameter
Method
Precision
Significant
Minimum Digits
Detection , Deswed
Salinity
Temperature
Induction Salmometer" dtration:
or titratlon, (APHA, 1989)
Bathythermograph or - "i 0.05'C
Thermometri&i EPA
Method 170,1. |APHA, 1989}
Ippt
-i* \ - ••- •- U s
The water column, being a 3-D m&liuttu reguires greater numbers of samples to be
collected (vs. a 2-D medium) in oriieY to ^adequately described.
drift. It is recommended that duplicate determinations be made for at least 10 percent
of the samples analyzed (U.S. EPA, 1987a).
Many multi-probe w situ measurement systems incorporate depth measurements by
use of pressure transducers. The accuracy and precision of such system must be
periodically checked. A pressure calibration can be done in a laboratory pressure
chamber. Daily calibration may be done in calm waters of a known depth (e.g., inside
a lagoon or a marina). Precision is determined by multiple measurements at the same
depth. Accuracy is evaluated by comparison to measurements made with a heavily
weighted line.
Instruments that measure turbidity require regular calibration. In situ instruments are
most accurately calibrated by the use of primary standards, usually a carefully pre-
pared suspension of formazin or other approved standard. Transmissometer type
instruments can be calibrated by comparison to solutions of known total suspended
solids. These solutions should approximate the turbidity of the water that is to be
sampled. For other instruments, including those nephelometers or turbidity meters that
-------
B-27
V
measure discrete samples, either on deck or in the laboratory, calibration using an
approved primary standard suspension is recommended (APHA, 1989). Calibration in
all cases should be conducted at the start of each series of analyses and after each
group of ten successive samples. Duplicate analyses should be conducted on at least
ten percent of the total number of samples (U.S. EPA, 1986-1991).
Furthermore, the following recommendations should be considered:
• current meters should be calibrated before and after each major deployment
of instruments
• UNESCO (1988) presents details on the calibration of CTD instruments
• fluorometer calibration prior to use with a series of prepared concentra-
tions, blanks and spikes are recommended quality control checks (Wilson,
1986).
B1.5 In general, one expects to find temporal and spatial patterns in physical characteristics.
Statistical Design The primary purpose of monitoring the physical characteristics of the water column is
Considerations to describe these patterns (e.g., range, seasonal variations) and to determine the
primary physical mechanisms affecting the monitored area.
As a result, statistical comparisons between sampling locations and sampling periods
are not common except for turbidity. In this instance, the reader is referred to the
Water Chemistry section for analytical approaches that would be appropriate.
Temporal Analyses
Tidal and seasonal variability in the distribution and magnitude of several water
column physical characteristics are typically observed (Day et al., 1989). Time series
analyses - e.g., temporal autocorrelation, spectral analyses - may be necessary to
examine the effects of these cyclical influences and/or to filter out these cyclic forcing
functions in order to examine long-term temporal trends.
Graphical Representations
The physical characteristics data described in this section are typically summarized in
graphical form; the most common being temperature-depth profiles, salinity-depth
profiles, and temperature-salinity plot (T-S plot). Alternatively, horizontal sections
(e.g., contour maps) of properties are useful for displaying geographical distributions.
-------
B-28
WM^fljittiMMiiif MMMF ffraww « v
fcr inifu !iji i" 1 l.tfM
Bflplllllllllllllllllpl Illl I I I I 1NJ" *$* !f
StlllPllllllflllLIIIHlllllfllllllhll INK •* jfl
H)>l|^^w4WSHIW
-------
B-29
•\
Monitoring Design Considerations
• Temperature, salinity, density, turbidity, and depth (depth of sample and
depth to bottom) should be collected at a sufficient number of depth
intervals to obtain a realistic picture of the water column profile for tem-
perature and salinity at each sampling location; in most cases, aim
interval will provide the necesssary resolution
• Sampling frequency should be determined by the estuary program's
requirement for detecting seasonal or high frequency events (e.g. tides,
storms, etc.); measurements should be collected over a period of not less
than two years in order to examine seasonal effects (Fischer et al, 1976)
• More frequent sampling in fish and shellfish areas, and other biologically
sensitive areas may be required
Existing Analytical Methods
• Temperature measurements may be made with:
- mercury-filled centigrade thermometer
- thermistor
- reversing thermometer
- bathythermograph
- CTD instrumentation
• Salinity measurements are preferably made with conductivity cells (CTD
instruments)
Density measurements are to be calculated from salinity, temperature, and
pressure
Depth measurements are preferably made with hydrostatic pressure tech-
niques
Turbidity measurements may be made with:
- light transmissometer
- Secchi disc
- nephelometer
- turbidimeter
-------
B-30
ii »ii pit j
• Cument measurements may be made with:
- Eulerian methods - e.g., propeller-type meters, electromagnetic
meters, acoustic-doppler meters, etc.
- Lagrangian methods - e.g., drogues, surface drifters, etc.
QAIQC Considerations
• Calibrate all equipment regularly
• Blank and replicate analyses are recommended QC checks
Statistical Design Considerations
• The primary purpose is to describe what the patterns (e.g., range, seasonal
variations) of physical characteristics are and to determine the primary
physical mechanisms affecting the monitored area
• Statistical comparisons between sampling locations and sampling periods
are not common except for turbidity
• Common graphical summaries include:
- temperature-depth, salinity-depth profiles, and temperature-
salinity (T-S) plots
- horizontal sections (i.e., contour maps) of properties with mul-
tiple physical characteristics displayed by placing one graph
above the other
• Current data can be displayed with stick plots, a time series of rectangular
components, and horizontal maps (Pickard and Emery, 1982)
Use of Data
• Physical characteristics are used to determine hydrodynamics and to
provide ancillary information to interpret other variables
• Turbidity can be used to estimate the reduction of light transmittance. Reduc-
tion of the depth to which sunlight penetrates, due to turbidity increase, can
reduce biological community growth (Thomann and Mueller, 1987)
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B-31
B1.8 APHA. 1989. American Public Health Association, American Water Works Associa-
Literature Cited tion, and Water Pollution Control Federation, Standard methods for the examination of
and References water and wastewater, 17th Edition. American Public Health Association, Washing-
ton, D.C.
Appell, G.F., and W.E. Woodward. 1986. Proceedings of the IEEE Third Working
Conference on Current Measurement. Institute of Electrical and Electronic Engineers,
New York.
Appell, G.F., and T.B. Curtin. 1990. Proceedings of the IEEE Fourth Working
Conference on Current Measurement. Institute of Electrical and Electronic Engineers,
New York.
Becker, D.S. and J.W. Armstrong. 1988. Development of regionally standardized
protocols for marine environmental studies. Mar. Poll. Bull. 19(7)310-313.
Brainard, E.G. and R.J. Lukens. 1975. A comparison of the accuracies of various
continuous recording current meters for offshore use. Offshore technology confer-
ence, Paper No. OTC 2295.
Clausner, J.E., W.A. Birkemeier, and G.R. Clarke. 1986. Field comparison of four
nearshore survey systems. Miscellaneous Paper CERC-86-6, US Army Engineer
Waterways Experimental Station, Vicksburg, MS.
Davis, R.E. 1988. Modeling eddy transport of passive tracers. J. Mar. Res., 45,635.
Day, J.W., C.A.S. Hall, W.M. Kemp, and A. Yanez-Arancibia. 1989. Estuarine
Ecology. New York, NY: John Wiley & Sons. 558pp.
Demmann, W.P., J.R. Proni, J.F. Craynock, and R. Fargen. In Press. Oceanic waste-
water outfall plume characterization measured acoustically.
Dustan, P., and J.L. Pinckey, Jr. 1989. Tidally induced estuarine phytoplankton
patchiness. Limnol. Oceanogr. 34:410-419.
Eugene, L.B. 1966. Handbook of Oceanographic Tables. Naval Oceanographic
Office, Rep. SP-68.
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B-32
in i i in ui MM 1*1 m
Fofonoff, N. P. and R.C. Millani, Jr. 1983. Algorithms for Computation of Funda-
mental Properties of Seawater. United Nations Educational, Scientific, and Cultural
Organization, UNESCO, Technical papers in marine science #44 Paris, France.
Goldstein, RJ. 1983. Fluid Mechanics Measurements. New York, NY: Hemisphere
Publ. Corp.
Grace, R.A. 1978. Marine Outfall Systems Planning. Design, and Construction.
Englcwood Cliffs, NJ: Prentice Hall, Inc. 600pp.
Hill K.D., T.M. Dauphinee, and D. J. Woods. 1986. Extension of the practical salinity
scale 1978 to low salinities. IEEE Journal of Oceanic Engineering Vol OE-11, #1, Jan
1986, pp 109-112.
Johns, W.E. 1988. Near-surface current measurements in the Gulf Stream using an
upward-looking acoustic doppler current meters. J. Atmos. Ocean. Technol., 5, 602.
Kennish, M.J. 1989. CRC Practical Handbook of Marine Science. CRC Press, Inc.,
Boca Raton, FL.
Koh, R.C.Y. 1988. Shoreline impact from ocean waste discharges. J. Hydraul. Div.
Am. Soc. Civ. Eng. 114:361.
Leighton, J.P, 1988. Verification/Calibration of a thermal Discharge Model. Proc. Int.
Symp. Model-Prototype Correlation of Hydraul. Structures, 148.
McCullough, J.R. 1977. Problems in measuring currents near the ocean surface.
Oceans '77, Marine Tech. Soc. and Inst. of Electrical and Electronics Engineering.
Muellenhoff, W.P., A.M. Soldate, Jr., D.J. Baumgarthner, M.D. Schuldt, L.R. Davis,
and W.E. Frick. 1985. Initial mixing characteristics of municipal ocean discharges.
Volume I- Procedures and applications. EPA-600/3-85-073a. U.S. Environmental
Protection Agency, Narragansett, RI. 90 pp.
Pickard, G. L. and W. J. Emery. 1982. Descriptive Physical Oceanography. An
Introduction. 4th (SI) Enlarged Edition. New York, NY: Pergamon Press, Inc.
Pinckey, J., and P. Dustan. 1990. Ebb-tidal fronts in Charleston Harbor South Caro-
lina: physical and biological characteristics. Estuaries. 13:1-7.
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B-33
Pond S., and G. L. Pickard. 1983. Introductory Dynamic Oceanography. 3rd Edition.
New York, NY: Pergamon Press, Inc.
Riggs, N.P. and K.A. Thompson. In Press. Current Profilers: Comparing three
different techniques.
Thomann, R.V. and J.A. Mueller. 1987. Principles of Surface Water Quality Model-
ing and Control. New York, NY: Harper and Row Publ. 644 pp.
U.S. EPA. 1982. Design of 301(h) monitoring programs for municipal wastewater
discharges to marine waters. EPA 430/9-82-010. Washington, D.C.
U.S. EPA. 1983. Methods for chemical analysis of water and wastes, 2nd ed. EPA
600/4-79-020. U.S. Environmental Protection Agency, Environmental Support
Laboratory, Cincinnati, OH.
U.S. EPA. 1986. Quality Criteria for Water. Office of Water Regulations and Stan-
dards. EPA 440/5-86-001. Washington, D.C.
U.S. EPA. 1986-1991. Recommended protocols for measuring selected environmen-
tal variables in Puget Sound. Looseleaf. U.S. Environmental Protection Agency,
Region 10, Puget Sound Estuary Program, Seattle, WA.
U.S. EPA. 1987a. Quality Assurance/Quality Control (QA/QC) for 301(h) monitoring
program: Guidance on field and laboratory methods. EPA, Office of Marine and
Estuarine Protection. EPA 430/9-86-004. Washington D.C.
U.S. EPA. 1987b. Technical support document for ODES statistical power analysis.
EPA 430/9-87-005. Office of Marine and Estuarine Protection, Washington, D.C. 34 pp.
UNESCO. 1988. The acquisition, calibration, and analysis of CTD data. UNESCO
technical paper in marine science 54. France. 92 pp.
Wallace, J.W., and J. W. Cox. 1976. Design, Fabrication and System Integration of a
Satellite Tracked, Free-Drifting Ocean Data Buoy. NASA Technical Memorandum
X-72817, January, 1976.
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B-34
Wilson. 1986. Techniques of water-resources investigations of the United States
Geological Survey. Fluorometric procedures for dye tracing. Department of the
Interior, US Geological Survey. Washington, D.C.
Wright, SJ. 1988. Outfall Plume Dilution in Stratified Fluids. Proc. Int. Symp.
Model-Prototype Correlation of Hydraul. Structures, 148.
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B-35
B2.0 Water Column Chemistry
Government agencies and research institutions rely on monitoring data to evaluate the
quality of water bodies. However, there are a number of problems associated with
data collected over the years. One of the most significant problems is the lack of
comparability due to inconsistent choice of methods and poor technique and/or quality
control practices. Commonly employed EPA laboratory methods were intended to
detect pollutants at much higher concentrations than those usually found in natural
waters. Alternative analytical methods are at present being used or developed in
conjunction with existing NEP monitoring programs (e.g., Puget Sound, Chesapeake
Bay) and EPA methods continue to be updated. The best choice of individual analyti-
cal protocols will be dependent on site- or area-specific conditions, such as contami-
nants of concern, biological communities, and the level of anthropogenic impact. If
methods other than those approved by the EPA are required to achieve a desired
sensitivity, it should be demonstrated that these methods result in equivalent or better
data quality with respect to accuracy and precision. Several EPA documents (e.g.,
1987a, 1987b) discuss quality assurance/quality control (QA/QC) protocols, sampling
gear, and recommended laboratory methods. The Compendium of Methods for
Marine and Estuarine Environmental Studies (U.S. EPA, 1990), currently in prepara-
tion, includes a number of method variations for nitrogen species, phosphorus, and
chlorophyll.
B2.1 Estuarine waters represent an important habitat for many commercially, recreationally,
Rationale and ecologically important organisms. These waters also represent the medium in
which food, larvae, nutrients, and contaminants may be transported throughout the
estuary. The objective of monitoring water column chemistry is to detect and describe
spatial and temporal changes of contaminants in and around the estuary.
The results of monitoring of water column contaminant concentrations can provide a
spatial and temporal record of anthropogenic perturbations to the estuary. The results
may be used to monitor rates of recovery following environmental interventions, to
evaluate the condition of aquatic habitats, and to provide early warnings of potential
impacts to the estuarine ecosystem.
B2.2
Monitoring Design
Considerations
The decision of which chemicals to include in the monitoring program is not simple.
One approach is to request analysis for all chemicals on the EPA-defined Priority
Pollutant, Hazardous Substance, or Target Compound/Analyte Lists. However,
detection in the environment does not always correlate with biological effects. Fur-
thermore, many of the chemicals that pose risks to estuarine and marine systems are
hydrophobic. Water column monitoring for hydrophobic chemicals may indicate
-------
B-36
concentrations that are several orders of magnitude below concentrations in sediments
and biota. Unless compelling site-specific conditions warrant otherwise, water column
monitoring for hydrophobic chemicals is not recommended. Limitations in analytical
methodology, modelling techniques, and lexicological data restrict the usefulness of
the resulting data. In some cases, levels of contaminants which are of concern to the
health of organisms are below the lower limit of detection of the best available analyti-
cal methodology.
Selection of sampling locations and choice of sampling techniques are crucial deci-
sions to be made before the sampling effort is initiated. Random sampling may not
detect "hot spots" associated with point source discharges. Too much sampling close
to a point source discharge will, however, bias the evolution of overall ambient
conditions.
Water column sampling design must consider not only the horizontal location of the
sampling stations (addressed in Section 4.0), but also the vertical location within the
water column and the time of sample collection. Because the majority of chemical
analyses are performed, by necessity, on discrete samples, consideration should be
given to the number of depths at which samples are collected. Obviously, such
decisions will be based on site-specific characteristics, such as water depth, contami-
nants of concern, suspected sources, and the sensitivity or importance of local biota.
Costs related to the laboratory analyses should also be considered when determining
the number of samples desired.
The time of sample collection is a site- and target compound-specific consideration
which can strongly influence data comparability. For example, dissoved oxygen
concentrations fluctuate diurnally especially in shallow brackish water. Some con-
taminant concentrations may vary as a result of tidally driven freshwater or saltwater
intrusions. The effects of seasonal ambient wind patterns, or river flows and runoff
events associated with seasonal weather patterns should be considered in sample
design.
Water Column Sampling Equipment
Water column samples are frequently collected using water bottles. They are simple
devices, usually consisting of a cylindrical tube with stoppers at each end and a closing
device that is activated by an electrical signal. Each samples a discrete parcel of water
at any designated depth. Multiple samplers are fixed on a rosette frame in order that
several depths may be sampled during one cast and/or that replicate samples may be
-------
B-37
collected at a particular depth. The most commonly used bottle samplers include the
Kemmerer, Van Dom, Niskin, and Nansen samplers (U.S. EPA, 1986-1991). Alterna-
tively, a pump may be used to sample the water column.
Caged Organisms
The California Mussel Watch Program and the National Oceanic and Atmospheric
Administration (NOAA) Status and Trends Program have employed the use of caged
transplant mussels to monitor bioaccumulation of toxic chemicals over space and time
(Ladd et al., 1984; Goldberg et al., 1978). Caged sentinel species offer several advan-
tages:
they concentrate contaminants (up to lOlO fold vs. estuarine waters),
facilitating laboratory analyses of contaminant concentrations
they provide a means of temporally integrating water quality conditions
they provide an assessment of biological availability of water column
contaminants
• they may be deployed and maintained in a number of diverse locales
The Long Island Sound Estuary Program has shown that chemical residue analyses on
caged indicator species appears to be a promising approach for identifying sources of
pollution (U.S. EPA, 1982).
However, the disadvantages of caged organisms include:
• the cost of transplanting organisms
• the possibility of loss of the cage/buoy system
• the possibility of introducing a "nuisance" species
Species have different bioaccumulation potentials for various contaminants. It is
recommended that multiple sentinel species be deployed in order to ensure that a
number of contaminants are sufficiently evaluated and a comprehensive characteriza-
tion of water contaminants be conducted. Unfortunately, formulations which would
allow comparisons of bioaccumulation data between different species and/or different
-------
B-38
tissues types have not yet been developed; comparisons of tissue burdens among
species is not acceptable. Thus, it is essential that monitoring design elements be
standardized to allow for comparisons among estuarine studies.
Chemical Analyses
Questions to be considered during the choice of appropriate analytical methods include
the parameters of interest, desired detection limits, sample size requirements or restric-
tions, methods of preservation, technical and practical holding times, and matrix
interferences (especially from saline water). Saline waters cause additional problems
due to matrix effects. D'Elia et aL (1989) discuss the common analytical problems
encountered during monitoring analyses of water samples.
Selection of appropriate methods should consider a compromise between full-scan
analyses, which are economical but cannot provide optimal sensitivity for all com-
pounds, and alternate methods that are more sensitive for specific compounds, but can
result in higher analytical costs. The Chesapeake Bay program, for example, has used
alternative methods or modified EPA approved methods to achieve greater sensitivity
in the analyses of several nutrient and trace metal parameters. Possible interferences
include suspended solids, metal ions, and residual chlorine. A list of existing analytical
techniques is presented in Table B2-1.
Dissolved Oxygen - The titrimetric, or Winkler, method is the first method of choice
for the measurement of dissolved oxygen (U.S. EPA, 1983). The membrane electrode
method is recommended for samples containing interferents such as sulfur compounds,
chlorine, free iodine, color, turbidity, or biological floes, or when continuous monitor-
ing is planned (U.S. EPA, 1983).
Nutrients-Nutrients such as ammonia nitrogen, total Kjeldahl nitrogen, nitrate-nitrite
nitrogen, and total phosphorus are typically determined by spectrophotometric measure-
ments using a segmented continuous flow analyzer where samples and reagents are
continuously added in sequence separated by air bubbles and pumped through glass
tubing (U.S. EPA, 1983). Automated analyses can be accomplished in less time than
would be required by manual methods because each analysis is not carried to comple-
tion, but is brought to the same stage of development and exposure by the timing of the
stream flow through the system.
Ammonia nitrogen is determined by treating the samples with alkaline phenol and
hypochlorite to produce indophenol blue which is intensified with sodium
B2.3
Existing Analytical
Methods
-------
B-39
TABLE B2-1, LIST OF EXISTING ANALYTICAL TECHNIQUES
DISSOLVED OXYGEN
* Wihkler Titration
,,v. » Membrane Electrode
NUTRIENTS
,"" • Continuous Flow Spectrophotometry
Ammonia Nitrogen
Total KjeldaM Nitrogen
Jsfitrate-Nitrite Nitrogea
Total Phosphorus
» Fluorescence Spectrometry
Chlorophyll a
» High Performance Liquid
Chromatography (HPLQ
TRACE METALS
» Atomic Absorption Spectrophotometry (AA)
* flame
- graphite furnace
-coW vapor ,,
- gaseous hydride
» Inductively Coupled Plasnta Emission -
Spectrometry (ICP)
.QRGANICS
» Gas Chromatography (GC)
v > - with electron capture detection (GC/ECD)
~ - with mass spectrometry C/MS)
US, EPA Method 3<30,2
tWIj EPA Method 360.1 _
U,S. EPA Method 350.1
TJ.S, EPA Method 351,2^
U.S. EPA Method 3&.2
U.S, EPA Method 365,4
XJ,S. EPA Method 7000 Series
< EPA Method 7470 *
US. EPA Methods 7060,
and 7740
O,S,EPA Method 6010
, EPA Method 8Q§Q
ILS. EPA Methods 8240
,j, .and 8270
nitroprusside. Standard solutions should be made up using substitute sea water to
approximate the matrix of the samples (U.S. EPA, 1983). Total Kjeldahl nitrogen is
defined as the sum of free ammonia and organic nitrogen compounds which are
converted to ammonium sulfate under the conditions of digestion (U.S. EPA, 1983).
Sulfates react with nitrogen compounds of biological origin, but may not convert
nitrogenous compounds of some industrial wastes. Two digestion methods have been
used for determining Kjeldahl nitrogen, the "block method" (U.S. EPA Method 351.1)
and the "Helix method" (U.S. EPA Method 351.2). The block method is recom-
-------
B-40
mended because the Helix method underestimates Kjeldahl nitrogen due to incomplete
digestion of the sample. The Chesapeake Bay Program has developed adjustment
equations to compensate for these differences and increase accuracy when the Helix
method is used (Computer Sciences Corporation, 1992). For the determination of
nitrate-nitrite nitrogen, a filtered sample is passed through a granulated copper-
cadmium column to reduce any nitrate to nitrite. The nitrite is then transformed to a
highly colored azo dye which is measured spectrophotometrically.
The Chesapeake Bay Program has modified U.S.EPA Method 350.1 for ammonia
nitrogen determination by increasing the flow-through cell length to 50 mm and using
a different set of buffers. These modifications have provided the greater sensitivity
required for estuarine monitoring. This program also recommends the following set of
procedures to determine organic nitrogen rather than U.S. EPA Method 351.2 for Total
Kjeldahl Nitrogen: 1) total nitrogen is determined either by combustion using a CHN
analyzer, or by oxidation of all nitrogen forms to nitrate for determination as nitrate-
nitrogen, 2) analysis of nitrate, nitrite, and ammonia nitrogen by colorimetry, and 3)
calculation of organic nitrogen as the difference between the total nitrogen and the sum
of the nitrate, nitrite, and ammonia values.
Total phosphorus is determined by heating the sample in the presence of sulfates, then
cooling the sample and measuring phosphorous spectrophotometrically.
The fluorometric method for chlorophyll a is more sensitive than the spectrophotomet-
ric method. The sample is subjected to an excitation wavelength and the fluorescence
is measured at a second emission wavelength (APHA, 1989). The greatest uncertainty
in the method is the choice of reference standard. HPLC is the most accurate of the
methods used for the analysis of chlorophyll (since all forms can be separately quanti-
fied), however it is also the most expensive.
Trace Metals—The choice of analytical method for trace metal analysis is determined
by the required detection limit. ICP allows the simultaneous measurement of several
elements. However, the achievable detection limits are usually not as low as those
obtained by graphite furnace or hydride AA. The combination of AA and ICP is the
recommended analytical method for detection of metals since no technique is best for
all elements. Greater sensitivity in trace metal analyses can frequently be achieved
through extraction and concentration using chelation extraction. For example, al-
though cold vapor AA is the recommended technique for the analysis of mercury (U,S.
EPA, 1986-1991), greater sensitivity is achieved if mercury is first concentrated and
then analyzed by the gold amalgam/mercury technique.
-------
B-41
Graphite furnace A A is more sensitive than flame AA or ICP, but is more subject to
matrix and spectral interferences, which result in potential QC problems during the
analyses and uncertainties in the resulting data. Because of the lower concentrations
which can be detected by graphite furnace AA, particular caution must be taken with
regard to laboratory contamination. The concentration of each element is determined by a
separate analysis, making the analysis of a large number of contaminant metals both labor-
intensive and relatively expensive compared to ICP. However, AA methods may be cost-
effective for the analysis of a few metals over a large number of samples.
Semivolatile Organic Compounds - Analysis of semivolatile organic compounds
involves a solvent extraction of the sample, cleanup of the extract, GC separation, and
quantitation (U.S. EPA, 1986-1991). There are two GC/MS options for detecting
extractable organic compounds: internal standard technique and isotope dilution. The
isotope dilution technique is recommended because reliable recovery corrections can
be made for each analyte with a labeled analog or a chemically similar analog. This
method is, however, more expensive and less widely employed than the internal
standard technique, which is the current method of choice in the Contract Laboratory
Program. Mass spectrometry provides positive compound identification by compari-
son of both retention time and spectral patterns with standard compounds.
The identification of pesticides and PCBs can be made by GC/ECD analysis (U.S.
EPA, 1986-1991). GC/ECD provides greater sensitivity (lower detection limits)
relative to GC/MS, however GC/ECD does not provide positive compound identifica-
tion. Since the identity of a compound is determined solely by matching retention time
with that of a standard, confirmation of pesticides and PCBs on a second GC column is
required for confidence in the reliability of the qualitative identification of the com-
pound. When a compound is present at a high enough concentration, its identity
should be confirmed by the use of GC/MS.
Glass capillary GC achieves a much higher degree of resolution in the analysis of PCB
congeners than the standard packed column methods; however, few labs regularly use
this specialized technique.
Volatile Organic Compounds - Analysis of volatile organic compounds also in-
volves GC analysis and quantification (U.S. EPA, 1986-1991). The purge and trap
GC/MS technique is commonly employed for detecting volatile organic compounds in
water. As in the case of semivolatiles, GC/ECD may be used to achieve lower detec-
tion limits, although this introduces a level of uncertainty to the qualitative identifica-
tion of compounds. The isotope dilution technique is recommended if the DQOs of
-------
B-42
the monitoring program require accurate quantitation of each compound. This tech-
nique, however, carries additional analytical time and expense.
Sample Collection B2.4
QA/QC
Sampling Gear—The primary criterion for an adequate sampler is that it consistently Considerations
collect undisturbed and uncontaminated samples. Water column sampling devices
should be inspected for wear and tear leading to possible sample leakage upon ascent.
It is prudent to have a backup sampler onboard the survey vessel in case the primary
sampler is found to be unsuitable during the cruise.
In the field, sources of contamination include sampling gear, lubricants and oils,
engine exhaust, airborne dust, and ice used for cooling samples. Samples designated
for chemical analyses require all sampling equipment - e.g., siphon hoses, scoops,
containers - be made of noncontaminating material and be cleaned appropriately prior
to use. Potential airborne contamination - e.g., stack gases, cigarette sinoke - should
be avoided. Furthermore, samples and sampling containers should not be touched with
ungloved fingers.
Sample Handling and Storage
Splitting water samples for chemical analyses should only be conducted with
noncontaminating tools under "clean room" conditions. Recommended container
materials, sample sizes, preservation techniques, and storage lifetimes for all com-
pounds of concern in water are summarized in Table B2-2.
Laboratory Analyses
Laboratory performance as measured by method QA/QC protocols should be used to
evaluate and select appropriate analytical methods. Changes to laboratory protocols
should only be considered if new protocols meet established performance criteria and
provide accuracy and precision equivalent to or greater than EPA approved methods.
Suggested QA/QC protocols and analytical performance criteria are discussed below.
QA/QC reports should describe the results of quantitative QA/QC analyses, as well as
other elements critical to the laboratory analyses of water, sediments, and tissues to
ensure proper interpretation of the results. It is recommended that these reports be
recorded and stored in a database for future reference.
-------
B-43
TABLE B2-2* ^ ~
SAMPLE PRESERVATION A^ STORAGE PARAMETERS
Storage ,,
Analyte
Container1
Preservative Lifetime
Nutrients
Ammonia
Nitrate
Nitrate + Nitrite
Nitrite
Organic
Totalor dissolved metals
(except Hg)
Total or dissolved Hg
Partfculate Metals
P,G
500ml
100ml " I
jrefrigerate
200ml H2SO4pH<2^ 2Sdays
100ml >
500ml
refrigerate
11 ,, HNO3pH<2 6 mo
2SQml s HN<>3pH<2 8 days
a P as linear polyethylene, G - Itorosilicate glass, T^B = tetmfluoroethylene^-
b it i$ recommended that samples be analyzed as soon aspossibte,
& IF aliquot for Hg tafcen from this i litersample, cannot ose linear polyethylene,
3 Samples should be filtered as soon as possible awJ always within 24 h. Workshop
attendees recommend that filtering be done shipboard rafter than hi lab on shore.
Field QA/QC Checks - Travel (trip) blanks will indicate whether any contamination
occurred in the field or during shipping of samples. Rinsate blanks check for contami-
nation due to inadequate cleaning of field equipment
-------
B-44
<
Held splits, treated and identified as separate samples, may be sent to the same labora-
tory for analysis or one sample sent to a "reference" laboratory for comparison.
Standard reference material may be placed in a sample container at the time of collec-
tion and sent "blind" to the laboratory.
Instrument QA/QC Checks - Calibration standards should be analyzed at the begin-
ning of sample analysis, and should be verified at the end of each 12-hour shift during
which analyses are performed (U.S. EPA, 1987b). The concentrations of calibration
standards should bracket the expected sample concentrations, otherwise sample
dilutions or sample handling modifications (e.g., reduced sample size) will be required.
Method QA/QC Checks- Analysis of preparation blanks should be conducted to
demonstrate the absence of contamination from sampling or sample handling in the
laboratory. At least one method blank must be included with each batch of samples
and should constitute at least 5% of all samples analyzed.
Spike recovery analyses are required to assess method performance for the particular
sample matrix. Spike recoveries serve as an indication of analytical accuracy, whereas
analysis of standard reference materials measure extraction efficiency. Recommended
control limits include 75-125 percent recovery for spikes, and 80-120 percent recovery
for the analysis of standard reference materials.
Replicates must be analyzed to monitor the precision of laboratory analyses. A
minimum of 5% of the analyses should be laboratory replicates. The control limits are
±20 percent variation between duplicates. Triplicates should be analyzed on one of
every 20 samples or on one sample per batch if less than 20 samples are analyzed.
Laboratory performance and calibration should be verified at the beginning and end of
each 12-hr shift during which analyses are performed.
Currently, there exists no universally agreed upon convention for reporting detection limits
of analytical procedures. Table B2-3 lists definitions of various detection limits used by
the American Chemical Society's Committee on Environmental Improvement (CEI). The
IDL does not address possible blank contaminants or matrices interferences and is not a
good standard for complex environmental matrices. The LOD and LOQ account for
blanks, but not matrix interference. The MDL provides high statistical confidence but,
like the LOQ, may be too stringent. Detection limits must be specified and reported to
ensure adequate data quality and comparability among protocols.
Detection limits should be low enough, as practically possible, to allow detection of
contaminants of concern and subsequent statistical analyses.
-------
B-45
B2.5
Statistical Design
Considerations
DEFINITIONS FO& SELECTED OMITS OP DETECTION,
Instrument Detection Limit (IDL) the smallest signal above Sjackgroami noise that an
iftstrunient can detect reliably* „
Hmit of Detection (LOD) , the lowest cmicenttation fevel that can be tfeteaained
' to l)e^te(istlcaIlydiffeient%HniEhe blank. The
/ecommeft
-------
B-46
\ v A v
-------
B-47
-iw.
Monitoring Design Considerations
• The objectives of the monitoring program must be clearly defined in order
that:
- the analytical methods show appropriate selectivity, specificity
and sensitivity for the contaminants of concern
- sampling and sample handling procedures are well-defined and
consistent so as not to compromise the integrity and representa-
tiveness of the samples
- sampling numbers and locations are appropriate to the level of
information required
• Water column monitoring for hydrophobic chemicals is not recommended.
• Time of sampling can strongly influence data comparability.
Existing Analytical Methods
• Dissolved Oxygen
- The Winkler (titrimetric) method is the first method of choice for
the measurement of dissolved oxygen, unless the samples contain
interferents such as sulfur compounds, chlorine, free iodine,
color, turbidity, or biological floes, or when continuous monitor-
ing is planned. In those instances, a membrane electrode should
be used
• Nutrients
- nutrients such as ammonia nitrogen, total Kjeldahl nitrogen,
nitrate-nitrite nitrogen, and phosphorus are typically determined
by spectrophotometric measurements using a segmented continu-
ous flow analyzer
- the usual method for chlorophyll determination is fluorometric,
however the choice of method is determined by the need to
separate the different forms. HPLC is the most accurate method,
however, it is also the most expensive
• Trace Metals
- a combination of AA spectroscopy and ICP spectrometry is the
recommended method for the detection of metals
-------
B-48
- cold vapor AA spectroscopy is the recommended protocol for the
detection of mercury
• Organics
- for monitoring efforts where the most accurate quantitation is
important, the isotope dilution GC/MS method is recommended
for volatiles and semivolatiles. GC/ECD is the method of choice
for pesticides and PCBs
QAIQC Considerations
• Calibration standards and blank, matrix spike, and replicate analyses are
recommended quality control checks
• A report describing the objectives of the analytical effort, methods of
sample collection, handling and preservation, details of the analytical
method, problems encountered during the analytical process, any necessary
modifications to the written procedures, and results of the QC analyses
should be included with the quantitative data
Statistical Design Considerations
• The analyst typically estimates statistics for central tendency (e.g., mean,
median) and variability (e.g., standard deviation, interquartile range) of
grouped data. Uncertainty should be indicated by reporting estimates with
confidence limits or percentiles
• For the purpose of estimating summary statistics, it is generally recom-
mended not to use left censored data
• Prior to the collection of data, the statistical test (and significance level)
used to analyze the data should be specified
Use of Data
• Monitor ambient levels of pollutants in the environment
• Establish spatial and temporal trends in the accumulation and transport of
pollutants discharged into ambient waters
-------
B-49
Calibrate and verify mathematical models
Calculate nutrient budgets
Develop water quality standards for receiving waters
Identify noncompliant discharges
B2.8 APHA. 1989. American Public Health Association, American Water Works Associa-
Literature Cited tion, and Water Pollution Control Federation, Standard methods for the examination of
and References water and wastewater, 17th Edition. American Public Health Association, Washing-
ton, D.C.
Armstrong, J.W. and A.E. Copping. 1989. Comparing the Regional Puget Sound
marine monitoring with the NOAA National Status and Trends Program. Coastal
Zone Proceedings 3:2421-2435.
Becker, D.S. and J.W. Armstrong. 1988. Development of regionally standardized
protocols for marine environmental studies. Mar. Poll. Bull. 19(7):310-313.
Bergstrom, P. 1990..Chesapeake Bay Coordinated Split Sample Program Annual
Report, 1989. Chesapeake Bay Program, Annapolis, M.D.
Chesapeake Bay Program. 1991. Coordinated Split Sample Program Implementation
Guidelines, Revision 3. EPA Chesapeake Bay Program, Annapolis, M.D.
Computer Sciences Corporation. 1992. Adjusting helix Kjeldahl nitrogen results:
Maryland Chesapeake Bay mainstem water quality monitoring program, 1984-1985.
Final Report to the Chesapeake Bay Program, U.S. EPA Region III.
D'Elia, C.F., P. A. Steudler, and N. Corwin. 1977. Determination of total nitrogen in
aqueous samples using persulfate digestion. Limnol. Oceanogr. 22:760-764.
D'Elia, C.F. et al. 1987. Nitrogen and phosphorus determinations in estuarine waters:
A comparison of methods used in Chesapeake Bay monitoring. Final Report to the
Chesapeake Bay Program, U.S. EPA Region III. US Government Printing Office,
Washington, DC.
-------
B-50
D'Elia, C.F. ef al. 1988. Nutrient analytical services laboratory standard operating
procedures. Chesapeake Biological Laboratory, University of Maryland.
D'Elia, C.F., J.G. Sanders, and D.G. Capone. 1989. Analytical chemistry for environ-
mental sciences: A question of confidence. Environ. Sci. Technol. 23(7).
Gilliom, RJ. and D.R. Helsel. 1986. Estimation of distributional parameters for
censored trace level water quality data 1. Estimation Techniques. Water Res. Res.
22(2):135-146.
Glibert, P.M. and T.C. Loder. 1977. Automated analysis of nutrient seawater:
Manual of techniques. Woods Hole Oceanographic Institute Technical Report No.
WH01-77-47.
Goldberg, E.D., V.T. Bowen, G.H. Farrington, J.H. Martin, P.L. Parker, R.W.
Risebrough, W. Robertson, E. Schneider and E. Gamble. 1978. The mussel watch.
Environ. Conserv. 5:101-125.
Hirsch, RM, 1988. Statistical methods and sampling design for estimating step trends
in surface-water quality. Water Res. Bull. 24(3):493-503.
Knezovich, J.P. and F.L. Harrison. 1987. A new method for determining the concen-
tration of volatile organic compounds in sediment interstitial water. Bull. Environ.
Contain. Toxicol. 38:837-940.
Ladd, J.M., S.P. Hayes, M. Martin, M.D. Stephenson, S.L. Coale, J. Linfield, and M.
Brown. 1984. California state mussel watch: 1981-1983., Trace metals and synthetic
organic compounds in mussels from California's coast, bays and estuaries. Biennial
Report. Sacramento, CA: Water Quality Monitoring Report No. 83-6TS.
Plumb, R.H. 1981. Procedure for handling and chemical analysis of sediment and
water samples. Technical Report EPA/CE-81-1. U.S. EPA and Corps of Engineers,
U.S. Army Engineers Waterways Experimental Station, Vicksburg, MS.
Porter, P.S., R.C. Ward, and H.F. Bell. 1988. The detection limit. Env. Sci. Tech.
22:856-861.
Salley, B.A., J.G. Bradshaw, and BJ. Neilson. 1986. Results of comparative studies
of preservation techniques for nutrient analysis on water samples. Virginia Institute of
Marine Science Report to the Chesapeake Bay Liaison Office, September 1986.
-------
B-51
Strickland, J.D.H. and T.R. Parsons. 1968. A Practical Handbook of Seawater
Analysis. BuU. Fish. Res. Board Can. 167:310p.
Tetra Tech. 1986. Recommended protocols for measuring selected variables in Puget
Sound. Final report submitted to Puget Sound Estuary Program by Tetra Tech, Inc.
Bellevue, Washington.
U.S. EPA. 1979. Handbook for analytical quality control in water and wastewater
laboratories. EPA 600/4-79-019. Environmental Monitoring and Support Laboratory,
Cincinnati, OH.
U.S. EPA. 1982. Methods for organic chemical analysis of municipal and industrial
wastewater. EPA 600/4-82-057. Environmental Monitoring and Support Laboratory,
Cincinnati, OH.
U.S. EPA. 1983. Methods for chemical analysis of water and wastes, 2nd ed. EPA
600/4-79-020. U.S. Environmental Protection Agency, Environmental Support
Laboratory, Cincinnati, OH.
U.S. EPA. 1986. Test methods for evaluating solid waste. EPA Publication SW-846,
3rd Edition. Office of Solid Waste and Emergency Response, Washington, DC.
U.S. EPA. 1986-1991. Recommended protocols for measuring selected environmen-
tal variables in Puget Sound. Looseleaf. U.S. Environmental Protection Agency,
Region 10, Puget Sound Estuary Program, Seattle, WA.
U.S. EPA. 1987a. Quality Assurance/Quality Control (QA/QC) for 301(h) Monitor-
ing Programs: Guidance on field and laboratory methods. EPA 430/9-86-004. Office
of Marine and Estuarine Protection, Washington, DC.
U.S. EPA. 1987b. Technical support document for ODES statistical power analysis.
EPA 430/9-87-005. Office of Marine and Estuarine Protection, Washington, DC.
34pp.
U.S. EPA. 1990a. Statement of work for organics analysis: multi-media, multi-
concentration. Document no. OLMO 1.0. U.S. Environmental Protection Agency,
Contract Laboratory Program, Washington, DC.
-------
B-52
U.S. EPA. 19905. Compendium of methods for estuarine and marine environmental
studies. EPA 503/2-89-001. U.S. Environmental Protection Agency, Office of Water.
Washington, D.C.
U.S. EPA. 1991. Statement of work for inorganic analysis: multi-media, multi-
concentration. Document no. BLM02.0. U.S. Environmental Protection Agency,
Contract Laboratory Program, Washington, DC.
Valdcrama, J.C. 1981. The simultaneous analysis of total nitrogen and total phospho-
rus in natural waters. Mar. Chem. 10:109-122.
Ward, R.C. and J.C. Loftis. 1986. Establishing statistical design criteria for water
quality monitoring systems: Review and synthesis. Water Res. Bull. 22(5):759-767.
-------
B3.1
Rationale
B-53
B3.0 Sediment Grain Size
The objective of monitoring sediment grain size composition is to detect and describe
spatial and temporal changes of the benthic environment. The availability of sediment
contaminants are often correlated with the grain size composition of the benthic
medium; sediment contaminants are more easily adsorbed onto small grain sediment
surfaces. Likewise, grain size information may explain the temporal and spatial
variability in biological assemblages; changes in sediment grain size often affect an
infaunal organism's ability to build tubes, capture food, and escape predation. These
results may be used to monitor rates of recovery following environmental interven-
tions, to evaluate the condition of benthic habitats, and to assist in providing early
warnings of potential impacts to the estuarine ecosystem.
B3.2
Monitoring Design
Considerations
Sediment Sampling Devices
Sampling crews have been given a wide latitude in how sediment samples are collected. In
fact, the protocols required to collect an acceptable surficial sediment sample for subse-
quent measurement of physical characteristics have generally been neglected. However,
because sample collection protocols influence all subsequent laboratory and data analysis,
it is key that sediment samples be collected using acceptable and standardized techniques.
Collection of undisturbed sediment requires that the sampler:
• create a minimal pressure wave when descending
• form a leakproof seal when the sediment sample is taken
i
i
• prevent winnowing and excessive sample disturbance when ascending
• allow easy access to the sample surface in order that undisturbed
subsamples may be taken
Penetration well below the desired sampling depth is preferred to prevent sample
disturbance as the device closes. It is preferable to use a sampler that has the means of
weight adjustment in order that penetration depths may be modified.
Numerous types of devices can be used to collect sediment samples. These can be
divided into two general categories: grabs and box corers (Mclntyre et al., 1984;
ASTM, 1991). Many of these devices sample the benthic habitat in a unique manner
(Table B3-1). Accordingly, conducting comparisons among data collected using
different devices is uriadvisable.
-------
B-54
PISWVwj.*^
1.L\?W"<* W1W-
•s ^Sx&X^v.''., >X*v;
TABLE B3-1 SUMMARY OFBOTTOM SAMPLING' EQUIPMENT3
Device
', Advantages
Disadvantages
Fluorocarbon plastic or
Glass Tube
Hand Corer with
removable Fmorocar-
bon plastic or glass
liners
Box corer
Gravity corers, that is,
Phlcger Corer
Young Grab (fiuoidear-
bon plastic- or kynar-
Mned modified 0,1m2
van Veen)
Ekman or Box Dredge
Shallow wadeable waters or
deeVwaters if SCUBA -
available:; Softdtsemi-" "„
corisolidaied deposits,
Same as above except more
consolidated sediments %can
be obtained, ^ ,
Same as above.
$emi-cons6lidated sedi-
ments, , S-.
Preserves layering and
permits historical Study of
sediment deposition, fiapid-
samples imtnediately ready
for laboratory shipment.
Minimal risk of contamina-
Small sample sizseiequfres
repetitive sampling.
tion.
Handles provide for greater
s ease of substrate penetration.
Above advantages.
Collection of large undis*
turbed sample allowing for
subsampling;
Low risk: of Cample contamk
nation. Maintains sediment
integrity relatively well.
Careful handling necessary to
prevent spillage. Requires
removal of liners before
Lakes and marine areas.^ .. Eliminates metal contamina-
^t^v^ ._ ""*** %tjon.. Reduced pressure
-'- - wave-
Soft Iff semi-soft sediments. , Obtains a larger sample than
Can be usedfrom boat, N coring tubes. Can be
bridge| Or pierXin waters of subsampled through box lid.
various dejpflist " " * -s!<
PONAR Grab Sampler
, HXS «
Useful on sah^silt, or clay. Most universal grab sampler,
_ - ""-^ - Adequate on most sub-
\x ""%v ^ strates. Large sample
, " s^ '"" -Obtainedintact,permitting
tsV- , " ,\ % N JUbsampJing,
frdm barrel and core cutter.
Hard to handle*
Careful handling necessary to
avoid sediment spillage.
Small sample, requires
repetitive operation and
removal of Hnets, Time
consuming,
Expensive. Requires winch.
Possible incomplete jaw
closure and sample loss.
Possible shock wave which
may disturb the fines, Metal
construction may introduce
contaminants. Possible loss
of "fines" on retrieval.
Shock wave from descent
may disturb "fines", Possible
incomplete closure of jaws
results in sample loss.
Possible contamination from
metal frame construction.
Sample must be further '
prepared for analysis.
-------
B-55
Device
•>TABLE B35L SUMMARY OF BOTTOM SAMPLING EQUIPMENT8
Advantages Disadvantages
Use
^fJ Piston Corer" s Wafers of 4-6 ft deep when
, „ - used w& extension jtod.
- -^ ;Soft to jemi-eonsolidated
on sand, $ihy6r £lay*
Piston provides for greater
sample retention.
Adequate on most sul>- "
strates; Large sample <
- q^tamed intact, permitting
$ub$arnpBr)g< s ^
site to other containers-metal,
barrels introduce risk of
metal contamination.
Shock wave from descent TO,
may disuirb "fines" Possible
incomplete closure ofja#s
results in sample loss, ,
Possible contamination from
metat frame construction.
Sampling moving waters
% from ^fixed platform,
^ \ % V.
Petesrsen Grab Sampler"is XJ^ful on most sabstrates:
Shipek<5rab Sampler • Usedprimarify in rrianne
, ^ % -" "watersandlargelmland
lakes and reservoirs.
Orange-Peel Grab ^^ - -XJsefol on most sabstrates*
Smiui-Mclntyre Grab
allowsTslmpling where othejr
Devices could notachieve-
« ipropei" orientation* "
---4--V m,^ - t ~~
* ™^ " - ,0
Large sample; can penetrate
most substrates. ~?,
Sample bucket ma/ be
opened to permit subsam-
pling. -Retains fine grained
sediments effectively, x<'
Designed for sampling hard
Substrates* \
, Sam
prepared for analj(|Js<
' •• •^'W
Possible eontamtoation ffoni
metal cflnstrufition, Subsam-
pling dffilpalts Not effecUve
for sanipiing fine sediments*
•-
v may require mm&.
"Nocoverlid to permit
subsampfing. All other
disadvantages of fifcrnati and
"'
Possible contarninafioB from
metal constructiorj,
may require winch.
-Loss of. floes* HeaV|~
require winch, Possime ^
metal contamination,
Scoops, Drag Buckets
Various environments
endittg on depth and
Inexpensive^ easy B handle. Lossoffmesonrettieval ,
~- dirough water coluawi.
-substrate.
'•* Ooinmentstepresent subjective evaluations.
SOURCE; American Society for Testing and Materials, 1991
-------
B-56
Grab samplers - Grabs are capable of consistent sampling of bottom habitats. De-
pending on the size of the device, areas 0.02 to 0.5 m2 and depths ranging from 5 to 15
cm may be sampled. Limitations of grab samplers include:
• variability among samples in penetration depth depending on sediment
properties
• oblique angles of penetration which result in varying penetration depths
within a sample
• the sample is inevitably folded, resulting in the inability to section the
sample and the loss of information concerning the vertical structure in the
sediments
However, the careful use of these devices will provide quantitative data. Grab sam-
plers are the tools of choice for a number of estuarine and marine monitoring programs
due to their ability to provide quantitative data at a relatively low cost (Fredette et al,
1989; U.S. EPA, 1986-1991).
Core samplers—Box corers utilize a surrounding frame to ensure vertical entry;
vertical sectioning of the sample is possible (U.S. EPA, 1987a). These devices are
capable of maximum penetration depths of 15 cm and may collect volumes 5 to 10
times that of grab samplers. Limitations of box corers include:
• its large size and weight require the use of cranes or winches and a large
vessel for deployment
• higher construction expenses
• lack of calibration studies to permit comparisons to grab samples
The Hessler-Sandia box corer uses dividers to section the core into subsections,
facilitating subsampling of the core. Box corers are recognized as the tools of choice
for maximum accuracy and precision when sampling soft bottom habitats.
Sediment Profiling Camera - The sediment profiling camera allows vertical in situ
imaging of the water-sediment interface from which grain size determinations may be
conducted. The sediment grain size determinations may be made at a maximum depth of
18 cm, however, penetration depth of the viewing prism may be limited due to the physi-
cal characteristics of the sediment (i.e., penetration depths are greater in silt than sand).
-------
B-57
Distinction between finer silts and clays are not possible. Furthermore, it is recom-
mended that quantitative calculations of sediment grain size from the image be verified
by the results of laboratory analysis of collected sediment samples.
The singular use of the sediment profiling camera is not recommended. However, the
sediment profile camera proves to be effective as a reconnaissance tool. Delineation
of habitats with similar physical characteristics may aid in the selection of appropriate
sampling stations.
Recommendations - Collection of sediments and benthic organisms should be done
concurrently in order to mitigate the costs of field sampling and to permit sound
correlation, regression, and multivariate analyses (see Benthic Community Structure).
Therefore, it is recommended that the sampling device also be suitable for benthic
sampling. Grab and core sampling devices permit adequate sampling of both sediment
and benthic infaunal communities with the one sampling device.
Sample Depth
It is recommended that the upper 2 cm of the sediment column be examined to charac-
terize surficial sediments. Although the 2 cm specification is arbitrary, it will ensure
that:
• relatively recent sediments are sampled
• adequate volumes of sediments are readily obtained for laboratory analyses
• data from different studies may be compared
Sampling of depths other than 2 cm or vertical stratification of deeper sediment cores
may be appropriate, depending upon the objectives of the monitoring program and the
rate of sediment accumulation. For example, if information concerning only the most
recent sedimentation events is required, examination of the upper 1 cm may be appro-
priate. Stratification of deeper cores will provide historical data of sediment grain size
and depositional events. If the potential for bioaccumulation of contaminants in
infaunal organisms is a concern, sampling to the depth of the anoxic layer is desirable.
Comparison of data from studies analyzing different sediment depths is not advised.
Penetration well below the desired sampling depth is preferred to prevent sample
disturbance as the device closes.
-------
B-58
Selection of Time of Sampling
Sediment grain size compositions are often temporally stable, although some slight
seasonal variability may be present. Changes are usually associated with seasonal
patterns of benthic turbulent mixing and sediment transport phenomena. The fre-
quency of sampling should be related to the expected rate of change in grain size
compositions. A consistent sampling period is recommended in order that spatial and
temporal comparisons may be conducted.
If seasonal variations are exhibited, it is recommended that direct comparisons be-
tween samples collected during different seasons be avoided. Studies investigating
intcrannual variation in the percent composition of grain sizes should be conducted
during the same season (preferably the same month) each year. Furthermore, it is
recommended that grain size be sampled when contaminant concentrations are ex-
pected to be at their highest level in order to evaluate worst-case scenarios.
Grain Size
Sediment grain size may be expressed in either mm or § (phi) units. These scales are
related according to the equation: = -Iog2 (mm). Data should be converted to phi
units before calculation of grain size parameters. Sediments are broadly classified into
three size classes: silts and clays are less than 0.064 mm (4 (j>) in diameter, sands range
from 0.064 mm (4 <|)) to 1 mm (0
-------
B-59
SWA
grain size analysis will allow all comparisons between samples within each study and
among different studies.
Sieving
Sieving separates the sediment sample into size fractions. Wet sieving separates the
sample into two fractions: grain size greater than 62.5 um (i.e., sand and gravel) and
less than 62.5 um (i.e., silt and clay). Aggregates should be gently broken and wet
sieving should continue until only clear water passes through the sieve.
The gravel-sand fraction can be further divided by dry sieving through a graded series
of screens (U.S. EPA, 1986-1991). The sample should be shaken on the mesh for a
pre-determined, standard amount of time. The last sieve should have a mesh size of
62.5 um (4 phi) and any material passing through this sieve should be added to the silt-
clay fraction of that sample. The silt-clay fraction can be subdivided using a pipet
technique (U.S. EPA, 1986-1991). This technique depends upon differential settling
rates of different sized particles. Withdrawal time for pipet analysis, as a function of
particle size and water temperature, is given in Table B3-2. The total weight of each
phi-size interval must then be calculated (U.S. EPA, 1986-1991).
An alternate method for the determination of grain size for the sand fraction of the
sediment is the use of a settling tube. This technique is based on Stake's Law which
describes the sinking rate of a particle relative to its diameter. This technique requires
a much smaller sample size (0.5-1.0 gm) as compared to 100 gm required for dry
sieving. Furthermore, settling tube analysis is relatively rapid, and automated settling
tubes which input the data directly to a personal computer are available.
It is key that the analytical techniques and the desired number of subfractions be
specified and standardized to allow for comparisons between samples.
B3.4
QA/QC
Considerations
It is critical that each sample be homogenized thoroughly in the laboratory before a
subsample is taken for analysis. Laboratory homogenization should be conducted
even if samples were homogenized in the field as sediments will differentially sort
themselves during transport and handling. In addition, after dry-sieving a sample all
material must be removed from the sieve.
The total amount of fine-grained material used for pipet analysis should be 5-25
grams. If more material is used, particles may interfere with each other during settling
and the possibility of flocculation may be enhanced. If less material is used, the
experimental error in weighing becomes large relative to the sample size. Once the
-------
B-60
TABLE B34. SEDIMENT GRAIN SIZE;
WITHDRAWAL TIMES FOR PIPET ANALYSIS AS A FUNCTION OF
PARTICLE SIZE AND WATER TEMPERATURE3^
Diameter Diameter
Finer Finer Withdrawal
than than Depth ' '
(phi)c (urn) (cm) ,18*C
Elapsed Time for Withdrawal of Sample in
Hours (h), Minutes (m), and Seconds (s)
arc
4.0
5.0
6.0
7.0
9.0
10.0
62.5
31.2
15.6
7.8
3.9
1.95
0.98
20
10
10
10
10
10
10
20s
2mOs
8mOs
20s
4m57s
^ S
7m48s
20s
Im54s
20s
Im51s
7m25s
20s
Im49s
-20s
Im46s
31m59s Slmlls 30m26s 29m41s 28m59s
•> 's^ ////^/ s
2h8m ^2h5m 2h2m Ih59m' ' th56m
> *<.« ff s s
8h32ms 8hl8m 8h6m 7h56m , 7h.44m ' ?h32m
34h6m ' 33hl6m 32h28m 31h40m 30h56m 30hl2m
• Modified from Plumb 0981),
b It 5s critical that temperature be held constant during the pipet analysis.
c <{>» -Iog2 ^particle diameter (mm)]. * s s ,
d Breakpoint between silt and clay.
20s
Ib51m
7h22m
pipet analysis begins, the settling cylinders must not be disturbed, as this will alter
particle settling velocities.
It is recommended that triplicate analyses be conducted on one of every 20 samples, or
on one sample per batch if less than 20 samples are analyzed. It is recommended that
the analytical balance, drying oven, and temperature bath be inspected daily and
calibrated at least once per week.
-------
B-61
w*. ff sSs %.
B3.5 Statistical strategies may mitigate the high costs of collecting sufficient quantities of
Statistical Design sediment. See also Statistical Design Considerations: Composite Sampling, Power
Considerations Analysis, and Power Cost Analysis (Section B.3).
B3.6 Sediment grain size provides evidence essential in the evaluation of spatial and tempo-
Use Of Data ral effects of anthropogenic and natural disturbances.
Grain size information may explain the temporal and spatial variability in biological
assemblages. Because many infaunal organisms are sensitive to changes in grain size
(e.g., affects their ability to burrow, to construct tubes), changes in benthic community
structure may be explained by simply examining grain size characteristics.
The detection of a change in sediment grain size composition may provide an early
warning of a potential threat to sensitive resources, such as a threat to commercial
clam beds. For many commercially and recreationally important clams, the ability to
quickly burrow, and to escape predation, is dependent upon the sediment grain size
composition of the bottom. Thus, changes of sediment grain size may be a good
indicator of impending loss of important resources.
However, the singular use of sediment grain size to assess the condition of the benthic
habitat or to guide the decision-making process is not recommended. The data gar-
nered from monitoring of the sediment's physical characteristics should be used in
conjunction with that collected from chemical and biological monitoring. Subsequent
analyses of health risks to human populations, as well as ecological risks to benthic
individuals, populations, and communities may be evaluated.
B3.7
Summary and
Recommendations
Rationale
The objective of monitoring the sediment's physical characteristics is to
detect and describe spatial and temporal changes in these characteristics.
Results may be used to: monitor rates of recovery following environmen-
tal interventions; assist in interpreting the results of physical and biological
monitoring programs; evaluate the condition of benthic habitats, and;
provide early warnings of potential impacts to the estuarine ecosystem.
-------
B-62
Monitoring Design Considerations
• It is recommended that consistent types of sampling gear, sampling loca-
tion, and timing of sample collection be implemented to allow for compari-
sons among studies
• Collection of undisturbed sediment requires that the sampler:
- create a minimal pressure wave when descending
- form a leakproof seal when the sediment sample is taken
- prevent winnowing and excessive sample disturbance when
ascending
- allow easy access to the sample surface in order that undisturbed
subsamples may be taken
• In general, analysis of the upper 2 cm will allow examination of recent
deposiUonal events; however, site-specific conditions and program objec-
tives should be considered when a sampling depth is selected
• Penetration well below the desired sampling depth is preferred to prevent
sample disturbance as the device closes.
Existing Analytical Methods
• Because true and apparent distributions differ, detailed comparisons
between samples analyzed by these different methods are questionable
• It is therefore desirable that all samples within each study and among
different studies be analyzed using the same method (i.e., either including
or excluding organic material)
QA/QC Considerations
• Laboratory homogenization should be conducted even if samples were
homogenized in the field
• It is recommended that the analytical balance, drying oven, and tempera-
ture bath be inspected daily and calibrated at least once per week
-------
B-63
\\
•NXw.
Statistical Design Considerations
• Compositing sediment samples consists of mixing two or more field
replicates or field samples collected at a particular location and time
• Space- (combining composites from several locations) and/or time- (com-
bining several composites over time from one location) bulking strategies
should be used judiciously since information concerning spatial and
temporal heterogeneity may be lost
• Power analyses have shown that for a fixed level of sampling effort, a
monitoring program's efficiency is generally increased by collecting more
replicates at fewer locations
Use of Data
• Grain size information may explain the temporal and spatial variability in
biological assemblages
• The singular use of sediment grain size to assess the condition of the
benthic habitat or to guide the decision-making process in not recom-
mended; data garnered from monitoring of the sediment's physical charac-
teristics should be used in conjunction with that collected from chemical
and biological monitoring
B3.8 ASTM. 1991. Standard guide for collection, storage, characterization, and manipulation
Literature Cited of sediments for lexicological testing. ASTM designation E1391-90. In: Annual book of
and References ASTM standards. American Society for Testing and Materials, Philadelphia, PA.
Buller, A.T. and J. McManus. 1979. Sediment sampling and analysis. In: Estuarine
hydrography and sedimentation. (K.R. Dyer, ed.) Cambridge: Cambridge University
Press.
Folk,R.L. 1974. Petrology of Sedimentary Rocks. Austin, TX: Hemphills. 170pp.
-------
B-64
444M* ^ ^ l \ tjyvi «,
^
<
Frcdcttc, T.J., D.A. Nelson, T. Miller-Way, J.A. Adair, V.A. Sotler, J.E. Clausner,
E.B. Hands, and F. J. Anders. 1989. Selected tools and techniques for physical and
biological monitoring of aquatic dredged material disposal sites. Final Report. U.S.
Army Engineer Waterways Experiment Station, Vicksburg, MS.
Mclntyre, A.D., J.M. Elliot, and D.V. Ellis. 1984. Introduction: design of sampling
programs. In: feTethods for the Study of Marine Benthos. IBP Handbook No. 16.
(N.A. Holme and A.D. Mclntyre, eds.). Oxford: Blackwell Scientific Publications.
pp. 1-26.
Plumb, R.H. 1981. Procedure for handling and chemical analysis of sediment and
water samples. Technical Report EPA/CE-81-1. U.S. EPA and Corps of Engineers,
U.S. Army Engineers Waterways Experimental Station, Vicksburg, MS.
Shcpard, P.P. 1954. Nomenclature based on sand-silt-clay ratios. J. Sed. Petrol.
24(3):151-158.
U.S. EPA. 1986-1991. Recommended protocols for measuring selected environmen-
tal variables in Puget Sound. Looseleaf. U.S. Environmental Protection Agency,
Region 10, Puget Sound Estuary Program. Seattle, WA.
U.S. EPA. 1987b. Bioaccumulation monitoring guidance: Strategies for sample
replication and compositing, vol. 5. EPA 430/9-87-003. Office of Marine and Estua-
rine Protection, Washington, DC. 51 pp.
U.S. EPA. 1987c. Technical support document for ODES statistical power analysis.
EPA 430/9-87-005. Office of Marine and Estuarine Protection, Washington, DC.
34pp.
-------
B-65
B4.1
Rationale
B4.0 Sediment Chemistry
The sediments represent the ultimate repository for many chemical contaminants in the
estuarine environment (U.S. EPA, 1992). Sediments also provide habitats for many
aquatic organisms. The objective of monitoring bulk sediment chemistry is to detect
and describe spatial and temporal changes of these sediment pollutants. The results
may be used to evaluate the condition of benthic habitats, to provide early warnings of
potential impacts to the estuarine ecosystem, and to monitor rates of ecosystem
recovery following environmental interventions.
Monitoring of pollutant levels in sediments is a widely accepted means of measuring
the condition of the benthic habitat and is a powerful tool for the evaluation of spatial
and temporal effects of anthropogenic and natural disturbances. The singular use of
sediment pollutant loading data to assess the condition of the benthic habitat or to
guide the decision-making process is not recommended since other factors, such as
water quality and sediment grain size, can also affect habitat quality.
B4.2
Monitoring Design
Considerations
Sediment Sampling Devices
The protocols required to collect an acceptable surficial sediment sample for subse-
quent measurement of chemical variables have generally been neglected. In fact,
sampling crews have been given wide latitude in determining how samples are col-
lected. However, because sample collection protocols influence all subsequent labora-
tory and data analysis, it is key that sediment samples be collected using acceptable
and standardized techniques.
Collection of undisturbed sediment requires that the sampler:
• create a minimal pressure wave when descending
• form a leakproof seal when the sediment sample is taken
• prevent winnowing and excessive sample disturbance when ascending
• allow easy access to the sample surface in order that undisturbed
subsamples may be taken (ASTM, 1991)
Penetration well below the desired sampling depth is preferred to prevent sample
disturbance as the device closes. It is optimal to use a sampler that has the means of
weight adjustment in order that penetration depths may be modified.
-------
B-66
Several types of devices can be used to collect sediment samples: dredges, grabs, and
box corers (Mclntyre et al., 1984). Each of these devices sample the benthic habitat in
a unique manner (Table B4-1). Accordingly, conducting comparisons among data
collected using different devices is inadvisable.
Grab samplers - Grabs are capable of consistent sampling of bottom habitats. De-
pending on the size of the device, areas of 0.02 to 0.5 m2 and depths ranging from 5 to
15 cm may be sampled. Limitations of grab samplers include:
• variability among samples in penetration depth depending on sediment
properties
• oblique angles of penetration which result in varying penetration depths
within a sample
• the sample is inevitably folded resulting in the inability to section the
sample and the loss of information concerning the vertical structure in the
sediments (U.S. EPA, 1991b)
However, quantitative data can be provided by the careful use of these devices. Grab
samplers are the tools of choice for a number of estuarine and marine monitoring
programs due to their ability to provide reliable quantitative data at a relatively low
cost(Fredette eta!., 1989; U.S.EPA, 1986-1991).
Core samplers—Box corers utilize a surrounding frame to ensure vertical entry;
vertical sectioning of the sample is possible (U.S. EPA, 1986-1991). These devices
are capable of maximum penetration depths of 15 cm and may collect volumes 5 to 10
times that of grab samplers. Limitations of box corers include:
• its large size and weight require the use of cranes or winches and a large
vessel for deployment
• higher construction expenses
• lack of calibration studies to permit comparisons to grab samples
The Hessler-Sandia box corer uses dividers to section the core into subsections,
facilitating subsampling of the core. Box corers are recognized as the tools of choice
for maximum accuracy and precision when sampling soft bottom habitats.
-------
B-67
TABLE B4-L SUMMARY OF BOTTOM SAMPLING EQUIPMENT!
Device
Use
Advantages
Disadvantages "«--•
Fluorooafbon plastic or Shallow wadeable waters or
Glass Tube. ^leep waters if SCUBA
s .available. Soft or semi-. -
consolidated deposits.
Han<| Corer with
^removable Fluorocar-
"bjjti plasjlcxxr glass
Jinets
Same as above except more
consolidated sediments can
" "be obtained.
Preserves layering and
permits historical study of
sediment deposition. Rapid-
samples immediately ready
for laboratory shipment
Minimal risk of contamina-
tion.
Handles provide fof greater!
ease of substrate penetration.
Above advantages, s
Small santple size requires
repetitive sampling.
Boxcorer
Same as above,
Gravity corers, that is, Semi-consolidated sedi-
Careful handling necessary to
prevent spillage, Requires
removaloflinerSbefore """
repetitive sampling, Slight s
risk of metal contamination,
from barrel and core cutter.
Hard to handle.
Young Grab (fluorocar- JJakes and marine areas.
bon plastic- orJcynar-
lined modified OJm2
van. Veen)
Ekman or Box Dredge Soft to semi-soft sediments,
Can be used from boat,.
bridge, or pier in waters of
various depths.
FQNAR Grab Sampler Useful on sand, silt, or clay.
turbed sample allowing for
subsampling.
Low risk of undisturbed
„ sample contamination
Maintains sediment integrity
relatively well
Eliminates metal contatnina-_
fion. Reduced pressure
waye.
Obtains a larger sample than
coring tubes^ Can be
Most universal grab sampler.
Adequate on most sub-
strates. Large sample
obtained intact, permitting
subsampling.
Careful handling ftecessaty to
avoid sediment spillage.
Small sample, requires
repetitive operation and
removal of liners, Time
consumtogx s. — -
Expensive. Requires winch;.,..,
Possible incomplete jaw
closure and sample loss.
Possible shock wave which:
may disturb the fines. Metal -
construction may fotroduce
contaminants. Possible loss -
ofL'fines" on retrieval.
Shock waye from descent
may disturb ^fiwss". Possible
incomplelie closure of jawsr v N s
results in sample loss. ^
Possible contamiaafio|K6t>ni"
metal frame consMiction. ••;;
^ Sampte^must ^farther,^ —«f
prepared for analysis, - % % ^ ^
-------
B-68
TABLE B4-I. SUMMARY OF BOTTOM SAMPLING EQUIPMENT*
; , (continued)
Device
Advantages
Disadvantages
BMH-53 Piston Corer
Van Veen
Waters of 4-"<> ft deep when
used with extension rod.
Soft to semi-consolidated
deposits, ;
Useful on sand, silt, or clay.
BMH-60
Sampling moving waters
from a fixed platform.
Peterson Grab Sampler Useful on most substrates.
Shipek Grab Sampler
Used primarily in marine
waters and large inland
lakes and reservoirs."
Orangc-Peel Grab Useful on roost substrates.
Smith-Mclntyre Grab " I
Scoops, Drag Buckets Various environments
depending on depth and
Substrate.
Piston provides for greater
sample retention.
Adequate on most sub-
-sjrates. Large sample
obtained intact, permitting
subsampling.
Streamlined configuration
allows sampling where other
devices could not achieve
proper orientation.
v Large sample; cart penetrate
most substrates.
^Sample bucket may; be
opened to permit subsam-
pling. Retains fine grained
sediments effectively, " -
'Designed for sampling hard
substrates, „,, , ,,
Inexpensive, easy to handle.
Cores must be extraded o»
Site to other containers-metal
battels introduce risk of
metal contamination.
Shock wave from descent
may disturb "fines". Possible
incomplete closure of Jaws
results in sample loss.
Possible contamination from
metal frame construction*
Sample must be further
prepared for analysis.
Possible contamination from
metal construction, Subsam-
pling difficult, Not effective
for sampling fine sediments.
Heavy, may require winch.
No cover lid to permit
subsampling, All other
disadvantages of Ekmari and
Ponar.
Possible contamination from
metal construction. Heavy,
rriay require winch.
Ix>ss of fines, Heavy-may
require winch. Possible
metal contamination,
Loss of fines on retrieval
through water column.
a Comments represent subjective evaluations.
SOURCE: American Society forTesting and Materials, 1991
-------
B-69
—•,
Recommendations - Collection of sediments and benthic organisms should be done
concurrently in order to mitigate the costs of field sampling and to permit sound
correlation, regression, and multivariate analyses (see Benthic Community Structure).
Therefore, it is recommended that the sampling device also be suitable for benthic
sampling. Grab and core sampling devices permit adequate sampling of both sediment
and benthic infaunal communities with the one sampling device.
Sample Depth
It is recommended that the upper 2 cm of the sediment column be examined to charac-
terize surficial sediments. Although the 2 cm specification is arbitrary, it will ensure
that:
• relatively recent sediments are sampled
• adequate volumes of sediments are readily obtained for laboratory analyses
• data from different studies may be compared
Sampling of depths other than 2 cm or vertical stratification of deeper sediment cores
may be appropriate, depending upon the objectives of the monitoring program and the
rate of sediment accumulation. For example, if information concerning only the most
recent sediment contamination is required, examination of the upper 1 cm may be
appropriate. Stratification of deeper cores will provide historical data of sediment
contaminant levels and depositional events. If the potential for bioaccumulation of
contaminants in infaunal organisms is a concern, sampling to the depth of the anoxic
layer is desirable. Comparison of data from studies analyzing different sediment
depths is not advised. Penetration well below the desired sampling depth is preferred
to prevent sample disturbance as the device closes.
Total Organic Carbon and Acid Volatile Sulfides Normalization
Total organic carbon (TOC) and acid volatile sulfides (AVS) are considered by many
to be the most important parameters in defining organic and metal concentrations in
sediments. Toxic hydrophobic contaminant concentrations have been found to be
related to the TOC content of the sediment (Karickhoff et al., 1979). Also, the concen-
trations of some divalent toxic metals have been found to be related to the AVS
concentration of the sediment (DiToro et al., in press). The AVS pool is available to
bind metals, thereby reducing their bioavailability.
-------
B-70
Total Organic Carbon CTOC) normalizations of organic contaminant concentrations
have been used to provide estimates of bioavailability. These normalizations appear to
account for some of the variability found in bioaccumulation rates and biological
community structure. TOC/lipid normalized accumulation factors (AF) have also been
used to predict tissue residue concentrations (Ferraro et al., 1990; Lake et al., 1987).
Normalization based on AVS concentrations is considered a potentially useful tool for
assessing the bioavailability of certain divalent metals (e.g., Cd and Ni). However,
only a few laboratories can perform these analyses at present and the procedure has not
undergone any round robin verification. In addition, sampling for AVS is very subject
to error as contact with air can greatly effect results. Although AVS normalizations
represent a promising tool for evaluation of sediment metals data, general use in
cstuarine monitoring programs may not yet be practical.
These normalizations assume the following:
• organic contaminants partition predominantly to sediment organic carbon;
metal contaminants partition predominantly to sediment AVS
i
• rapid steady-state kinetics of contaminants
Development of standardized methods for measuring sediment TOC and AVS in the
laboratory are required before normalized contaminant concentrations may be com-
pared among studies.
The decision to normalize for TOC and AVS will depend upon monitoring program
objectives. For example, if the objective is to identify the "footprint" of a discharge,
normalization may not be appropriate. However, if the monitoring objective is to
identify "hot spots" (i.e., those areas where bioavailable contaminants represent a risk
to human and ecological health), normalization may be justified.
Selection of Sampling Period
Sediment contaminant levels typically display temporal patterns due to seasonal
variability. These patterns are usually a result of seasonal changes in benthic turbulent
mixing and sediment transport phenomena. The frequency of sampling should be
related to the expected rate of change in sediment contaminant concentrations. A
spatially and temporally consistent sampling design (i.e., time of sampling event and
sampling locations do not change over time) is recommended in order that compari-
sons between areas over time may be conducted.
-------
B-71
If seasonal variations are exhibited, it is recommended that direct comparisons be-
tween samples collected during different seasons be avoided. Studies investigating
interannual variation in the concentrations of sediment contaminants should be con-
ducted during the same season (preferably the same month) each year. Furthermore, it
is recommended that sediments be sampled when contaminant concentrations are
expected to be at their highest level in order to evaluate worst-case scenarios.
B4.3
Existing Analytical
Methods
Questions to be considered during the choice of appropriate analytical methods include
the parameters of interest, desired detection limits, sample size requirements or
restrictions, methods of preservation, technical and practical holding times, and matrix
interferences. It will frequently be necessary to use methods other than those currently
approved by the EPA to achieve a desired sensitivity in an estuarine environment.
However, alternative methods should be considered only when they have been demon-
strated to provide a level of precision and accuracy equivalent to or exceeding that of
the EPA approved method. Any proposed changes in analytical methods should be
reviewed by analysts with experience in estuarine and coastal waters for applicability
to that specific estuary.
Several U.S. EPA documents (e.g., 1986a and 1986-1991) discuss the common
analytical problems encountered during analyses of sediment samples in monitoring
programs. Those estuary programs that have established monitoring programs may
provide an excellent source of information about analytical methods being developed
for estuarine systems.
Chemical Residue Analyses
Several factors determine achievable detection limits for a specific contaminant,
regardless of analytical procedure - a list of analytical procedures and U.S. EPA
method numbers is given in Table B4-2. These factors include:
• sample size; 50-100 g (wet weight) with a minimum final dilution volume
of 0.5 ml is considered adequate (U.S. EPA, 1986-1991)
• presence of interfering substances
• range of pollutants to be analyzed-the optimal method for one target
pollutant may not be optimal for others
-------
B-72
rmm fir ™M_»V " VfwmMfV !»&&. «**(« v^\ v m v^i
U *\- -"T-" -^^-^i -
u. ...,. * \. S.* . . . ' ' "'. '
• level of confirmation - qualitative (e.g., presence or absence) or quantita-
tive (e.g., residue concentrations) analyses
• level of pollutant found in the field and in analytical blanks
Selection of appropriate methods will be based on a trade-off between full-scan
analyses, which are economical but cannot provide maximum sensitivity for some
compounds, and alternate methods that are more sensitive for specific compounds but
can result in higher analytical costs.
Metals and Metalloids - Digestion methods for sediment samples are reviewed by
Plumb (1981). The U.S. EPA Contract Laboratory Program (CLP) requires the use of
HNO3:H2O2 (U.S. EPA, 1991a).
Trace element analyses by ICP (U.S. EPA method 6010) allows several elements to be
measured simultaneously. Detection limits of ICP for most metals are generally
comparable to those achieved by GFAAS; however, detection limits for some metals
(arsenic, selenium, and mercury) are significantly lower using AAS.
TABLE B4-2. LIST OF EXISTING ANALYTICAL TECHNIQUES
(ILS, EPA, I986a)""", " „ ,„' ' ^ "
METALS/METALLOIDS ^
• Atomic Absorption Spectrdphotdmetty (AAS) U.S, EPA Method 7000 series
- flame " " _ ,
- graphite furnace (GFAA) Sj- " ..-
- cold vapor * " U,S. EPA Method 7470
- gaseous hydride (HYDAAS) ^ U.S. EPA Methods 7Q60 and 7740
• Inductively Coupled Plasma Emission' s U.S. BPA Method 6010
Spectrometry (ICP) , ,
s *• ^
ORGANICS
• Gas Chromatography (GC)
- with electron capture detection (GC/EC£) U.S, EPA Method 8080
- with mass Spectrometry (GC/MS) \ U.S. EPA Methods 8240 and 8270
ICP- AI, Sb, As, Ba, Be, Bt Cd, Ca, Cr, Co, Cfl,Fe( Pb» Mg, Mn, Mot Ni, K, Se, S Fe, Pb, Mfe Mn, Hg, Mo, Hi, K, Se, Ag,
Na.Tl.Sn, V,andZn
-------
B-73
The combination of atomic absorption spectrophotometry (AAS, U.S. EPA Methods 7000
series) and inductively coupled plasma emission spectrometry (TCP) is the recommended
analytical method for detection of metals and metalloids since no technique is best for all
elements (U.S. EPA, 1986a). Cold vapor AAS (U.S. EPA method 7470) analysis is the
only recommended technique for mercury (U.S. EPA, 1986a).
Graphite furnace AAS (GFAAS) is more sensitive (i.e., lower detection limits) than
flame AAS, but is more sensitive to matrix interference and spectral influences -
GFAAS requires particular caution with regard to laboratory contamination and
consequently, GFAAS requires more skilled laboratory technicians. Both AAS
methods require that the concentration of each element be determined by a separate
analysis, making the testing of a large number of contaminant metals both labor-
intensive and relatively expensive compared to ICP. However, AAS methods may be
cost-effective when testing for a few metals over a large number of samples.
Semi-Volatile Organic Compounds - Analysis of semi-volatile organic compounds
involves a solvent extraction of the sample, cleanup of the characteristically complex
extract, GC analysis, and quantification (U.S. EPA, 1986a, 1987b). There are two GC/
MS options for detecting extractable organic compounds: internal standard technique
and isotope dilution. The isotope dilution technique, which requires spiking the
sample with a mixture of stable isotope labeled analogs of the analytes, is recom-
mended because reliable recovery corrections can be made for each analyte with a
labeled analog or a chemically similar analog (U.S. EPA, 1986a). The isotope dilution
method is more expensive and less widely employed than the internal standard technique,
which is the current method of choice in the Contract Laboratory Program (CLP).
Mass spectrometry provides positive compound identification by comparison of both
retention time and spectral patterns with standard compounds. However, this tech-
nique lacks the sensitivity to detect levels of most trace contaminants in estuarine
systems. Alternative methods providing greater sensitivity are available, but will be
compound specific and, therefore, more expensive.
The identification of pesticides and PCBs can be made by GC/ECD analysis (U.S.
EPA Method 8080). GC/ECD provides greater sensitivity relative to GC/MS, however
GC/ECD does not provide positive compound identification. Confirmation of pesti-
cides and PCBs on an alternative GC/ECD or preferably by GC/MS, when sufficient
concentrations occur, is recommended for reliable results (U.S. EPA, 1986a). The
detection and concentration of PCBs can be more accurately determined using com-
parison mixtures other than the standard industrial Aroclor mixtures. Evaluations of
-------
B-74
PCB assemblages in environmental samples by quantification as Aroclors or total
PCBs are of limited accuracy due to environmental degradation and differential
affinities of PCB congeners for different environmental compartments. A more
meaningful evaluation can be accomplished by quantification by PCB isomer group-
ings (dependent on the number of chlorine atoms per molecules). This has the advan-
tage of indicating the relative concentrations of the groups containing the most toxic
and bioaccumulating congeners (McFarland etal., 1986). An alternative method of
analysis is to test for the individual PCB congeners. Using congener-specific methods
instead of Aroclor standard mixtures provides more accurate identification and quantifica-
tion and eliminates the necessity of subjective decisions on the part of the analyst
(Eganhouse, 1990). However, beside being more expensive and time consuming, the
compatibility of these enhanced detection methods with the standard Aroclor method is an
important consideration for monitoring programs with existing historical data. All other
organic compound groups are recommended for analysis by GC/MS (U.S. EPA, 1986a).
Volatile Organic Compounds - Analysis of volatile organic compounds involves a
solvent extraction of the sample, cleanup of the complex extract, GC analysis, and quanti-
fication (U.S. EPA, 1986a and 1987a). The purge and trap GC/MS technique is employed
for detecting volatile organic compounds in sediments. GC/ECD may be used to achieve
lower detection limits, although identification of the compounds may not be as accurate.
A successful variation for detection of volatile organic residues in sediments involves a
device that vaporizes volatile organic compounds from the sediment sample under vacuum
and then condenses the volatiles in a super-cooled trap (Hiatt, 1981). The trap is then
transferred to a purge and trap device where it is treated as a water sample. The isotope
dilution method is recommended as it provides reliable recovery data for each analyte
(U.S. EPA, 1986a). However, this technique costs additional analytical time and expense.
Sample Collection
Sampling Gear - The primary criterion for an adequate sampler is that it consistently
collect undisturbed and uncontaminated samples. Sediment sampling devices should
be inspected for wear and tear leading to sample leakage upon ascent. It is prudent to
have a backup sampler on board the survey vessel in case the primary sampler is lost,
damaged, or found to be unsuitable during the cruise.
In the field, sources of contamination include sampling gear, lubricants and oils, engine
exhaust, airborne dust, and ice used for cooling samples. Samples designated for chemical
analyses require all sampling equipment - e.g., siphon hoses, scoops, containers - be made
of noncontaminating material and be cleaned appropriately prior to use. Potential airborne
B4.4
QA/QC
Considerations
-------
B-75
contamination - e.g., stack gases, cigarette smoke - should be avoided. Furthermore,
samples and sampling containers should not be touched with ungloved fingers.
Sample Condition - Benthic samples should satisfy the following sample acceptabil-
ity criteria (U.S. EPA, 1986-1991):
• sampler is not over-filled with sample so that the sediment surface is
pressed against the top of the sampler
• overlying water is present, indicating minimal leakage
• the overlying water is not excessively turbid indicating minimal sample
disturbance
• the sediment surface is relatively flat indicating minimal disturbance or
winnowing (Figure B4-1)
• the desired penetration depth is achieved
Figure B4-1.
Examples of acceptable
and unacceptable samples
(U.S. EPA, 1986-1991).
Acceptable if Minimum
Penetration Requirement Met
and Overlying Water is Present
Unacceptable
(Canted with Partial Sample)
Unacceptable
(Washed, Rock Caught in Jaws)
Unacceptable
(Washed)
-------
B-76
If the sample does not meet all the criteria, the sample should be rejected.
Sample Handling
For analyses of metals, samples should be frozen and kept at -20°C (U.S. EPA,
1987a). Although specific holding times have not been recommended by U.S. EPA, a
maximum of 6 months (8 days for mercury; ASTM 1991) would be consistent with
holding times for water samples (Tables B4-3 and B4-4).
TABLE B4-3.
SUMMARY OF SAMPLE COLLECTION AND PREPARATION
QA/QC REQUIREMENTS FOR ORGANIC COMPOUNDS
% ,, j ,
- ^ '_•• ' Maximum
Variable Sample Size8 .Container1* Preservation Holding Time
SemivotaUIes 50-100 g
Volatiles 40 ml
TOC 100ml
"G
G
Freeze "
Cool,40Ce
Phosphoric acid,
p%<2,
TVS
100-500 ml
under art inert
atmosphere
6 months0
14 days
'58 days
none
recommended
* Recommended field sample sizes for one laboratory analysis. If additional
laboratory analyses are required (i.e., tab replicates) the field sample size should be
adjusted accordingly.
u "" ' '""'''"'"
b G= Glass. j ,,.>,,,,, ":,,,., ,rTr^
^\\ '"J''f>-" , ' '". "" '" '
c This is a suggested holding time. No U^EPA criteria exist for thepreservation ,
of this variable. , ^
d No headspace or airpoCketS should renialn. ' \ ^ '^
c Freezing these samples will likely cause breakage 6'f the sample container, because
no airspace for expansion is provided.
-------
B-77
Table B4-4 " ^ > — %
SAMPLING CONTAINERS, PRESERVATION REQUIREMENTS, AND HOLDING TIMES
FOR SEDIMENT SAMPLES
Contaminant Container8 Preservation Holding Time
Acidity
Alkalinity
Ammonia
Sulfate
Sulfide
Sulfite ,
.Nitrate
.Nitrate-Nitrite
Nitrite "•
Oil and Grease
Organic Carbon
Metals
\Chromiutn VI
v... Mercury "^
Metals, except above
Organic Compounds
Extractables (including phthafetes,
nittosamines, organochforine pesticides,
PCBTs, nitroaromatics, isophorone,
Polynucfear aromatic hydrocarbons.
haloethers, chlorinated hydrocarbons
Extractables (phenols)
Purgeables (halocarbons and aromatics)
' v
Purgeables {acrofein andacryionitrile)
., '
Orthophosphate
Pesticides
*
Phenols
Phosphorus (elemental)
Phosphorus, total
Chlorinated organic compounds
V % •. '
a Polyethylene (P) or Glass (G)
P?G
P» G
P,G
P,G
P, G
P,G
P,G
P,G
P,G
G
P.G _
-
P,G
P'G
„
G, teflon-lined cap
GI teflon-lined cap
G, teflon-lined
septum
G, teflon-lined
septum
P,G
G, teflon-lined cap
P,G
G
P,G
G» teflon4ined cap
Cool,4'C
Cool44*C
Cool,44C
Cool,4'C
Cool, 4"C
Cool, 4*C .
Cool, 4*C
Cool,4'C^ ^
Cool,4*C
Cool, 4"C
Cool,4°C
Cool,4°C
^
Coolt4'C
_.
Cool,4eC
Cool,4sC
-
Cool^'C
Cool,4*C
Cool, 4°C
Cool, 4 'C
Cool^'C
Cool, 4'C
Cool,4'C
, ,
14 days"
""" 14 dajC T
28 days
28 days
2& days
48 hows
4& hours
28 days
48 hours
28 days1
28 days
" ,,40houtsm
-- " 8 days"™
6 months.
7 days, (until extraction)
30 days (after extraction)
« > , ^ - ---•-•-••••••
x
7 days (until extraction)
30 days (after extraci&m)
14 days
••
3 days , ,.
•.
48 hours'
7 days (until extraction)
30 days (after extraction)
28 days
48 hours
28 days
7 days (until extraction)
, 3&days (aftst extraction)
V.V.,.. -
SOURCE: American Society JbrTesting and Materials, 1991
-------
B-78
I
*. w.
For analyses of volatile compounds, samples should be stored in the dark at 4°C (U.S.
EPA, 1987a). Analyses of volatile compounds should be performed within 14 days of
collection as recommended by U.S. EPA (1990). If analyses of semivolatile com-
pounds will not be performed within the recommended 7-day holding time, freezing of
the samples at -20°C is advised. Holding times for frozen samples has not been
established by the EPA. A general guideline of a maximum of 6 months would be
consistent with water sample guidelines (U.S. EPA, 1987a).
Samples for determination of TOC or AVS should be analyzed as soon as possible. If
not analyzed immediately, TOC samples should be refrigerated and their pH brought
below 2 by addition of phosphoric acid. Acidification is recommended only when
inorganic carbon is below detection limits (APHA, 1990). Acid Volatile Sulfides
should be stored in airtight containers under an inert atmosphere and analyzed as soon
as possible.
Rescctioning sediment samples for chemical analyses should only be conducted with
noncontaminating tools under "clean room" conditions.
Laboratory Analyses
Laboratory performance as measured by method QA/QC protocols should be used to
evaluate and select appropriate analytical methods. Changes to laboratory protocols
should only be considered if new protocols meet established performance criteria.
QA/QC reports should describe the results of quantitative QA/QC analyses, as well as
other elements critical to the laboratory analyses of water, sediments, and tissues to
ensure proper interpretation of the results. It is recommended that these reports be
recorded and stored in a database for future reference.
Field QA/QC Checks - Travel (trip) blanks can indicate whether contamination was
introduced by reagents in the field or introduced during shipping of samples. Rinsate
blanks are designed to verify the absence of contamination that can be carried over
from one sample to another due to inadequate cleaning of field equipment.
Field splits, treated and identified as separate samples, may be sent to the same labora-
tory for analysis or one sample may be sent to a "reference" laboratory for comparison.
In addition, standard reference material may also be placed in a sample container at the
time of collection and sent "blind" to the laboratory.
-------
B-79
Instrument QA/QC Checks - Calibration standards should be analyzed at the begin-
ning of sample analysis, and should be verified at the end of each 12-hr shift during
which analyses are performed (U.S. EPA, 1987a). The concentrations of calibration
standards should bracket the expected sample concentrations, otherwise sample
dilutions or sample handling modifications (i.e., reduced sample size) will be required.
Method QA/QC Checks - Analysis of method blanks should be conducted to demon-
strate the absence of contamination from sampling or sample handling in the labora-
tory. At least one method blank must be included with each batch of samples and
should constitute at least 5% of all samples analyzed.
Spike recovery analyses are required to assess method performance for the particular
sample matrix. Spike recoveries serve as an indication of analytical accuracy, whereas
analysis of standard reference materials (SRM) measure extraction efficiency. Recom-
mended control limits include 75-125 percent recovery for spikes, and 80-120 percent
recovery for SRM.
Replicates must be analyzed to monitor the precision of laboratory analyses. A
minimum of 5% of the analyses should be laboratory replicates. Triplicates should be
analyzed on one of every 20 samples or on one sample per batch if less than 20
samples are analyzed. The acceptable variation among replicates is 20 percent or less.
Tables B4-5 and B4-6 provide a brief summary of QA/QC for laboratory analyses.
For information concerning detection limits, see Water Column Chemistry QA/QC
Considerations (Section B2.4).
B4.5 Statistical strategies may mitigate the high costs of collecting sufficient quantities of
Statistical Design sediment. See statistical design considerations: composite sampling, power analyses,
Considerations and power-cost analysis (Section B.3).
B4.6
Use of Data
The results may be used to monitor rates of recovery following environmental inter-
ventions, to evaluate the condition of benthic habitats, and to provide early warnings of
potential impacts to the estuarine ecosystem. The singular use of sediment pollutant
loading data to assess the condition of the benthic habitat or to guide the decision-
making process is not recommended. Data acquired from monitoring sediment
contaminant levels, in conjunction with the sediment's physical properties, may be
used to assess the bioavailability of these pollutants (SCCWRP, 1986). Subsequent
analysis of this information, in conjunction with biological data, may be used to assess
-------
B-80
•ft*
TABLE B4-5, SUMMARY OF QUALITY CONTROL SAMPLE
Analysis Type
Recommended Frequency of Analysis
Surrogate spikes
Method blank
Standard reference
materials
Matrix spikes
Spiked method blanks
Analytical replicates
Field replicates
% %
Requited in'every sample - minimum 3 neutral 2 acid spikes/
plus 1 spike for pesticide/PCB analyses^ and 3 spikes for
volatile Isotope dilution techniques (t.e.» with all available
labeled surrogates) i$ recommended for full scan analyses and
to enable recovery corrections to be applied to data*
One per extraction batch (semivolatile,organic$). One per
extraction or one per 12-hour shift, which ever is most frequent
(volatile organicsX " ~
<50 samples: one' per set of samples submitted to lab,
>50 samples: one per 50 samples analyzed.'
•> -.'•s
Not required incomplete isotope dilution technique used,
<20 samples: one per set of samples submitted to lab,
^20 samples:* 5 percent of total number of samples.
As many as required to establish confidence in method before,
analysis of samples
-------
B-81
TABLE B4-& SUMMARY OR WARNING
AND CONTROL LIMITS FOR QUALITY CONTROL SAMPLE
Analysis Type,
Recommended
Warning Limit
Recotttriiehded
Control Limit
Surrogate Spikes „
Method Blank
Phthalate*
Acetone
Other Organic
Compounds
Standard
Reference Materials
s 10 percent recovery
30 percent of the analyte
1 jig total or 5 percent
of the analyte
95 percent
confidence interval
Matrix spikes , 50-65 percent recovery
Spiked Method Blanks 50-65 percent recovery
Analytical Replicates
Field Replicates -
Ongoing Calibration
50 percent recovery
5 M&total or SO percent
- of the analyte
2.5 jig total or 5 percent
of tne analyte
95 percent confidence
Interval for Certified
Reference Material """"""•
50 percent recovery
50 percent recovery
coefficient of variation,
25 percent of
initial calibration
Washington State Department of Ecology (1991) has adopted sediment criteria and the
EPA has recently published a sediment classification methods compendium (U.S.
EPA, 1992). Both of these documents may provide useful guidance. Our ability to
extrapolate from sediment chemistry data to predictions of ecological or human health
risks is, however, greatly limited by our understanding of causal relationships between
exposure and response.
-------
B-82
* ^ sm
Rationale
• The sediments represent the ultimate repository for many anthropogenic
contaminants infiltrating the estuarine environment
• The objective of monitoring bulk sediment chemistry is to detect and
describe spatial and temporal changes of these sediment pollutants
Monitoring Design Considerations
• It is recommended that consistent types of sampling gear, and location and
timing of sample collection be implemented to allow for comparisons
among studies and overtime
• Collection of undisturbed sediment requires that the sampler:
- create a minimal bow wake when descending
- form a leakproof seal when the sediment sample is taken
- prevent winnowing and excessive sample disturbance when
ascending
- allow easy access to the sample surface in order that undisturbed
subsamples may be taken
• Penetration well below the desired sampling depth is preferred to prevent
sample disturbance as the device closes.
• In general, analysis of the upper 2 cm will allow examination of recent
sediment contamination events, however, site-specific conditions and
program objectives should be considered when a sampling depth is selected
• Total organic carbon normalization is recommended to allow comparisons
of chemical residue concentrations between locations - normalizing the
data will depend upon the objectives of the individual monitoring program
• It is recommended that sediments be sampled when contaminant concen-
trations are expected to be at their highest level
B4.7
Summary and
Recommendations
-------
B-83
Existing Analytical Methods
• It is recommended that consistent types of analytical protocols be imple-
mented to allow for comparisons among studies
• Laboratories should perform acceptably on Standard Reference Materials
and intercalibration exercises before performing routine analyses
• Periodic full-scan analyses should be undertaken to identify any new
pollutants which should be monitored
• Metals/Metalloids
- The combination of GFAA and ICP are the recommended
methods for the detection of metals and metalloids (U.S. EPA,
1987b)
- cold vapor AAS is the recommended protocol for mercury
detection
• Organics
- GC/MS in conjunction with isotope dilution is recommended for
the detection of semi-volatile organic compounds
- vacuum super-cooled trap in conjunction with a purge and trap
device is recommended for the detection of volatile organics
(Hiatt, 1981)
- isotope dilution option is recommended as it provides reliable
recovery data for each analyte (U.S. EPA, 1986a)
QA/QC Considerations
• The primary criteria for acceptable field sampling equipment and protocols
are the collection of undisturbed and uncontaminated environmental
samples
• Adequate sampling containers and preservation techniques must be used
and appropriate holding times followed to ensure the integrity of samples
and their analyses
• Blank, spike recovery, and replicate analyses are recommended quality
control checks
-------
B-84
• A report describing the objectives of the analytical effort, methods of
sample collection, handling and preservation, details of the analytical
methods, problems encountered during the analytical process, any neces-
sary modifications to the written procedures, and results of the QC analyses
should be included with the analytical data
Statistical Design Considerations
• Compositing sediment sampling consists of mixing samples from two or
more replicates collected at a particular location and time
• Space- (combining composites from several locations) and/or time- (com-
bining several composites over time from one location) bulking strategies
should be used judiciously since information concerning spatial and
temporal heterogeneity may be lost
Use of Data
• The data garnered from monitoring of the pollutant levels in estuarine
sediments should be used in conjunction with that collected from other
physical and biological monitoring to monitor rates of recovery following
environmental interventions, to evaluate the condition of benthic habitats,
and to provide early warnings of potential impacts to the estuarine ecosys-
tem
• The singular use of sediment pollutant loading data to assess the condition
of the benthic habitat or to guide the decision-making process in not
recommended
ASTM. 1991. Standard guide for collection, storage, characterization, and manipulation B4.8
of sediments for toxicological testing. ASTM Designation E1391-90. In: Annual book of Literature Cited
ASTM standards. American Society for Testing and Standards, Philadelphia, PA. and References
Bcrgstrom, P. 1990. Chesapeake Bay Coordinated Split Sample Program Annual
Report, 1989. Chesapeake Bay Program, Annapolis, MD.
Chesapeake Bay Program. 1991. Coordinated Split Sample Program Implementation
Guidelines, Revision 3. EPA Chesapeake Bay Program. Annapolis, MD.
-------
B-85
' W ^
DiToro, D.M., J.D. Mahony, D.J. Hansen, KJ. Scott, A.R. Carlson, and G.T. Ankley.
In Press. Acid volatile sulfide predicts the acute toxicity of cadmium and nickel in
sediments.
DiToro, D.M., J.D. Mahony, D.J. Hansen, KJ. Scott, M.B. Hicks, S.M. Mayr, and
M.S. Redmond. In Press. Toxicity of cadmium in sediments: the role of acid volatile
sulfide.
Eganhouse, R.P. 1990. Sources and magnitude of error associated with PCM measure-
ments. In: Southern California Coastal Water Research Project Annual Report 1989-
1990. Eds: J.N. Cross and D.M. Wiley. Southern California Coastal Water Research
Project, Long Beach, CA.
Ferraro, S.P., H. Lee, R.J. Ozretich, and D.T. Specht. 1990. Predicting bioaccumula-
tion potential: A test of a fugacity-based model. Arch. Environ. Contain. Toxicol.
19:386-394.
Fredette, T.J., D.A. Nelson, T. Miller-Way, J.A. Adair, V.A. Sotler, J.E. Clausner,
E.B. Hands, and F.J. Anders. 1989. Selected tools and techniques for physical and
biological monitoring of aquatic dredged material disposal sites. Final Report. U.S.
Army Engineer Waterways Experiment Station, Vicksburg, MS.
Hiatt, M.H. 1981. Analysis of fish and sediment for volatile priority pollutants. Anal.
Chem. 53:1541-1543.
Karickhoff, S.W., D.S. Brown, and T.A. Scott. 1979. Sorption of hydrophobic
pollutants on natural sediments. Water Res. 13:241-248.
Knezovich, J.P. and F.L. Harrison. 1987. A new method for determining the concen-
tration of volatile organic compounds in sediment interstitial water. Bull.
Lake, J.L., N.I. Rubinstein, and S. Parvignano. 1987. Predicting bioaccumulation:
Development of a partitioning model for use as a screen tool in regulating ocean
disposal of wastes. In: Fate and Effects of Sediment-bound Chemicals In Aquatic
Systems. (Dickson, K.L., A.W. Maki, and W.A. Brungs, eds). Florissant, CO: Sixth
Pellston Workshop.
Landrum, P.P. and J.A. Robbins. In Press. Bioavailability of sediment-associated
contaminants to benthic invertebrates. In: Sediments: Chemistry and Toxicitv of In-
Place Pollutants (Giesy, J.P., R. Baudo, and H. Muntau, eds). Lewis Publishers.
-------
B-86
McFariand, V.A., J.U. Clarke, and A.B. Gibson. 1986. Changing concepts and
improved methods for evaluating the importance of PCBs as dredged sediment con-
taminants. Miscellaneous Paper D-86-5. Department of the Army, Corps of Engi-
neers, Waterways Experiment Station, Vicksburg, MS.
Mclntyre, A.D., J.M. Elliot, and D.V. Ellis. 1984. Introduction: design of sampling
programs. IBP Handbook No. 16. In: Methods for the Study of Marine Benthos.
(N.A. Holme and A.D. Mclntyre, eds.). Oxford: Blackwell Scientific Publications.
pp. 1-26.
Plumb, R.H. 1981. Procedure for handling and chemical analysis of sediment and
water samples. Technical Report EPA/CE-81-1. U.S. EPA and Corps of Engineers,
U.S. Army Engineers Waterways Experimental Station, Vicksburg, MS.
SCCWRP. 1986. Polynuclear aromatic hydrocarbon contamination in sediments form
coastal waters of southern California. Southern California Coastal Water Research
Program, pp. 13-16.
U.S. EPA. 1986a. Analytical methods for U.S. EPA priority pollutants and 301 (h)
pesticides in estuarine and marine sediments. Prepared for the Office of Marine and
Estuarine Protection, Washington, D.C.
U.S. EPA. 1986b. Test methods for evaluating solid wastes, physical/chemical
methods. SW-846,3rd Edition. Environmental Protection Agency, Washington, D.C.
U.S. EPA. 1986-1991. Recommended protocols for measuring selected environmen-
tal variables in Puget Sound. Looseleaf. U.S. Environmental Protection Agency,
Region 10, Puget Sound Estuary Program, Seattle, WA.
U.S. EPA. 1987a. Quality Assurance/Quality Control (QA/QC) for 301(h) Monitor-
ing Programs: Guidance on field and laboratory methods. EPA 430/9-86-004. Office
of Marine and Estuarine Protection, Washington, D.C.
U.S. EPA. 1987b. Bioaccumulation monitoring guidance: Strategies for sample
replication and compositing, vol. 5. EPA 430/9-87-003. Office of Marine and Estua-
rine Protection, Washington, D.C. 51pp.
U.S. EPA. 1987c. Technical support document for ODES statistical power analysis.
EPA 430/9-87-005. Office of Marine and Estuarine Protection, Washington, D.C.
34pp.
-------
B-87
U.S. EPA. 1990. Statement of work for organics analysis: multi-media, multi-
concentration. Document no. OLM01.0. U.S. Environmental Protection
Agency, Contract Laboratory Program, Washington, D;C.
U.S. EPA. 1991a. Statement of work for inorganic analysis: multi-media,
multi-concentration. Document no. ILM02.0. U.S. Environmental Protection
Agency, Contract Laboratory Program, Washington, D.C.
U.S. EPA. 199 Ib. Evaluation of dredged material proposed for ocean disposal:
Testing Manual. EPA 503/8-91/001. Office of Water, Washington, D.C.
U.S. EPA. 1992. Sediment classification methods compendium. EPA 823-R-
92-006. Office of Water, Office of Science and Technology, Washington, D.C.
Washington State Department of Ecology. 1991. Sediment management
standards. Washington Administrative Code (WAC) Chapter 173-204. Olym-
pia, WA. 61 pp.
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B-88
-------
B-89
v-
B5.0 Plankton: Biomass, Productivity and
Community Structure/Function
B5.1 Although increased primary production resulting from intentional nutrient inputs has been
Rationale shown to increase fish stocks in some experimental systems, the possible increase in
fisheries in naturally productive systems is generally considered insignificant when
compared to the deleterious effects of eutrophication. The most conspicuous and poten-
tially detrimental effects of eutrophication to estuarine environments are increased turbid-
ity, phytotoxins, and creation of hypoxic or anoxic conditions. Furthermore, changes in
nutrient concentrations in an estuary may result in the potential for long-term biological
changes in the estuarine community structure and function (e.g., changes in species
distribution and abundance of both primary producers and consumers).
If eutrophic conditions are suspected of occurring, monitoring of the plankton community
may assist in detecting and defining the spatial and temporal scales of these conditions.
The objectives of monitoring plankton characteristics are to detect spatial and temporal
changes in plankton community structure and function. These results may be used to
assess the consequences of spatial and temporal variability in plankton community
structure and function to higher trophic levels which depend upon planktonic resources.
In addition, this information may be used to further the understanding of the relationship
between water quality conditions and planktonic community structure and function.
B5.2
Monitoring Design
Considerations
Sampling Methods
Phytoplankton - Plankton samples are frequently collected using water bottles. They
are simple devices, usually consisting of a cylindrical tube with stoppers at each end
and a closing device that is activated by a mechanical or electrical signal. Each
samples a discrete parcel of water at any designated depth. Multiple samplers are
fixed on a rosette frame in order that several depths may be sampled during one cast
and/or that replicate samples may be collected at a particular depth. The most com-
monly used bottle samplers include the Kemmerer, Van Dorn, Niskin, and Nansen
samplers (U.S. EPA, 1987a).
Phytoplankton samples should be collected at a variety of depths throughout the water
column, some above and some below the pycnocline. To adequately characterize the
phytoplankton community, it is recommended that a minimum of four depths in the
vertical profile should be sampled: 1) one meter below the surface, 2) one meter
above the bottom, 3) one meter above the pycnocline, and 4) one meter below the
pycnocline. If the waters are too shallow or no stratification occurs, it would be
appropriate to take the latter two samples at evenly spaced distances between the top
-------
B-90
and bottom samples. For example, composite water column samples collected for the
Virginia Chesapeake Bay Plankton Monitoring Program are taken at five different
depths above and five different depths below the pycnocline (U.S. EPA, 1990).
Vertical measures of chlorophyll a (an indirect measure of phytoplankton standing
stocks) can be made by fluorometric or spectrophotometric determination of chloro-
phyll a from these depth samples.
If available, a pump station may be used with a flow-through fluorometer for a con-
tinuous profile of chlorophyll a concentration with depth (Lorenzen, 1966). Samples
to be used for taxonomic analysis should be collected with water bottles, as agitation
associated with pumping may damage cells, making them unidentifiable. Pumps
should only be used for determination of chlorophyll a concentrations.
Zooplankton- Because zooplankton possess varying degrees of swimming ability,
they have the potential for aggregating in patches or in a narrow depth strata. This
introduces additional complications into quantitative sampling, such as avoidance of
certain types of gear.
Until recently, the role of the microbial loop, which consists of bacteria, flagellates, ciliates
and microzooplankton (<200 jam), has been overlooked. These organisms may represent a
significant pathway in the reutilization and conversion of dissolved organic carbon into
larger zooplankton and benthic organisms (Azam etal., 1983; Pomeroy, 1984).
The sampling methods to be used for collecting zooplankton will vary depending on the
size of the organisms. Microzooplankton (size range of 20-200 |jm) can be collected with
water bottles at various depths sinrilar to those used for phytoplankton, or small (for
example, 44 um) mesh nets can be used (Jacobs and Grant, 1978). Pumping systems can
also be used which have the advantage of being able to take samples integrated over depth
and of collecting samples while the ship is underway (Beers etal., 1967). However,
pumps may damage soft-bodied organisms and they are more expensive and complicated
than water bottles. Triplicate samples should be collected from each station depth, thus
allowing for statistical analysis of intra-station variability.
For small mesozooplankton (greater than 200 um) nets are generally used (UNESCO,
1968). Additional tows may have to be made with larger nets in order to collect
representative samples of larger zooplankton and larval fish. All tows should be
replicated. The number of replicates necessary for the desired precision of estimation
should be determined during a preliminary or pilot sampling program. A number of
other considerations, including net mouth diameter, towing speed and ship board
handling of samples will affect sampling results. Some problems associated with the
-------
B-91
W.V,
use of nets for zooplankton sampling include avoidance and clogging which may result
in underestimating abundance and diversity and loss of filtration efficiency (McGowan
and Fraundorf, 1966; Wiebe and Holland, 1968).
Conductivity-Temperature-Depth (CTD) Systems
Continuous vertical profiles of chlorophyll a concentrations can be obtained by
attaching a fluorometer to a conductivity, temperature and depth (CTD) system. An
additional sensor measuring dissolved oxygen could be attached to the CTD system.
This combined instrument package will make direct measurements of chlorophyll a
levels and the degree of eutrophication and water column stratification.
The advantage of this measurement system is that instead of pulling water up from
depth to a measuring device on the deck of a ship, the instrument is lowered through
the water column taking samples in undisturbed water. Sampling rates can be as high
as 24 samples per second with a descent speed of 1 meter per second. Data is usually
collected by a computer-based data acquisition system and in advanced systems can be
displayed in real-time as it is being collected. A rosette water sampler can be lowered
at the same time as the CTD package to collect water samples at discrete depths.
These samples will be used for taxonomy analysis and to verify and calibrate chloro-
phyll a measurements from the CTD fluorometer. The calibration will be carried out
using laboratory spectrophotometer methods.
Selection of Spatial and Temporal Sampling Strategies
The time of the year should be controlled or stratified in the design - the use of annual
averages is seldom good practice. Given the seasonal variation characteristic of
phytoplankton abundances, it is recommended that direct comparisons between
samples collected during different seasons be avoided. Studies investigating
interannual variation in the characteristics of phyto- and zooplankton communities
should be conducted during the same season (preferably the same month) each year.
Due to their short turnover times, phytoplankton communities are capable of respond-
ing to perturbations much more rapidly than other biotic groups. Therefore, phyto-
plankton samples should be collected relatively more frequently. In those situations
where phytoplankton communities display pronounced seasonal variations in standing
stock or production, it may be appropriate to use a temporally stratified sampling
approach. For example, in the Maryland Chesapeake Bay Phytoplankton Monitoring
Program, sampling takes place once monthly from October through March and twice a
month from April through September (U.S. EPA, 1990).
-------
B-92
The lifcspan of zooplankton on the other hand is longer than phytoplankton, so the
capacity for responding to perturbations is less than that of phytoplankton. Therefore
less frequent sampling is required. As with the phytoplankton community, the zoop-
lankton monitoring program should consider the natural temporal fluctuations in
abundance and species composition.
The distribution of planktonic organisms is strongly influenced by currents and tides.
Sampling programs should be developed to reflect the patchiness caused by tidal
fronts, and regions of convergence or divergence of currents within the estuary.
In many cases, regular monitoring of the zooplankton community may not be neces-
sary unless changes in the phytoplankton community are observed which would induce
changes in the herbivore community. Because many zooplankton graze on phyto-
plankton, in areas where the phytoplankton community has been affected, alterations
of the zooplankton community are a distinct possibility. In other cases, monitoring of
zooplankton may be desirable only when there is evidence of previous impact on the
zooplankton community, or in those situations where point and nonpoint source
discharges are located in areas where there is high potential impact on zooplankton.
For example, it may be useful to monitor zooplankton in estuarine environments with
macroplanktonic larvae of important commercial or recreational species.
Taxonomic Identification and Enumeration
Subsamplcs drawn from water collected in water sampling bottles should be preserved
for later microscopic analysis to determine phytoplankton community composition.
The choice of fixation will depend on the dominant types of phytoplankton known to
inhabit a given area (buffered formaldehyde and Lugol's solution are two common
fixatives). Preserved phytoplankton samples normally must be concentrated for
quantitative microscopic analysis.
The taxonomic analysis should include identification and enumeration of dominant
phyto- and zooplankton taxa and measures of population abundance whenever pos-
sible. Particular attention should be given to meroplanktonic larvae of commercially,
rccrcaUonally or ecologically important species.
The large number of species, incomplete taxonomy, and small size of the organisms
comprising the microbial loop make taxonomic identification extremely difficult.
Ecologists working with these groups tend to concentrate on higher taxonomic levels
and functional groupings. Staining with Acridine Orange, or similar stain, and direct
counting with epifluorescence microscopy has been used to estimate bacterial numbers
-------
B-93
\
(Harvey and Young, 1980; Wright and Coffin, 1983) and microprotozoan abundances
(Sherr and Sherr, 1983).
B5.3 Perturbations to the phyto- and zooplankton communities should be analyzed in
Existing Analytical relation to other potential impacts on other biological communities which include:
Methods
• potential primary and secondary impacts to higher trophic level communi-
ties (e.g., food web impacts)
• occurrence of toxic or nuisance phytoplankton
Some CCMPs may require that both larval and adult zooplankton community structure
and function should be analyzed in order to adequately assess impacts to high trophic
levels. Standardized methods are recommended to permit comparisons between
studies.
Biomass and Productivity
Methods for estimating phytoplankton biomass used in earlier monitoring surveys
include cell counts, total cell volume estimates, protein estimates, and dry weight.
These methods have certain disadvantages related to speed of the technique (e.g., time-
consuming) and the degree of accuracy for the estimation of phytoplankton biomass
(D'Elia et al., 1986). The use of chlorophyll a. measurements, especially fluorometric,
has become widespread primarily because the method is relatively fast, simple and
reproducible.
Phytoplankton biomass can be indirectly measured through the measurement of the
concentration of chlorophyll a in the water. This is done through fluorometric or
spectrophotometric measurements. Within these methods there are also differences in
extraction techniques for chlorophyll determination, including various methods of
filtration, solvents, temperature and/or physical treatment (sonication or grinding)
(D'Elia et al, 1986).
However, with both the fluorometric and spectrophotometric determinations, the
presence of accessory pigments may interfere with determination of chlorophyll a..
Several researchers, however, have successfully used chromatographic procedures to
separate interfering substances prior to determination (D'Elia et al., 1986). The High
Performance Liquid Chromatography (HPLC) is generally acknowledged as the most
-------
B-94
accurate (and most expensive) method for chlorophyll detenninations (Bidigare,
1989).
UNESCO (1973) recommends that the 14C light-dark bottle technique be used to
estimate primary production. This technique is more sensitive than, and requires
shorter incubation times than the O2 light-dark bottle method. However, the O2
method measures gross primary production, net primary production and respiration,
using inexpensive laboratory reagents, while the 14C technique estimates only net
primary production and requires specialized training and equipment, and relatively
expensive radioisotopes.
Biological Indices
Biological indices allow large amounts of multivariate data to be reduced to a single
number. The numbers of individuals and the numbers of species have been found to
be good indicators of anthropogenic disturbance, as well as of other environmental
stresses. Furthermore, these simple biological indices are less ambiguous and are often
as informative as diversity indices (U.S. EPA, 1985; Green, 1979; Hurlbert, 1971).
More complicated indices — e.g., species diversity, species richness, dominance,
evenness—have found varying degrees of acceptance. Diversity indices have the
following limitations (Green, 1984):
• often lack biological meaning
• are not robust empirical indicators of any important correlates of "estuarine
health"
• do not incorporate information of form and function of resident species
• are susceptible to biases associated with well-described taxa
However, species diversity indices are a widely used measure of community structure.
The dominance index is a measure of the degree to which one or a few species domi-
nate the community. The dominance index, herein defined as the minimum number of
species required to account for 75 percent of the total number of individuals, has been
useful in describing community structure (Swartz et al, 1985). It is easily calculated,
docs not assume an underlying distribution of individuals among species, and is
statistically testable.
-------
B-95
Indicator Species
Abundances of selected indicator species can be used to evaluate response of the
community as a whole. Examination of abundances of individual indicator species are
generally informative and may reduce the cost of the analysis. The absence of pollu-
tion sensitive species and the enhancement of opportunistic and pollution tolerant
species may assist in defining the spatial and temporal extent and magnitude of
impacts. However, indicator variables must possess the following characteristics
(Green, 1984):
• provide sufficiently precise and accurate appraisals of:
- species of concern
- anthropogenic disturbances to planktonic communities
- presence/absence or the magnitude of anthropogenic perturbance
to the estuarine ecosystem
• cost-effective and a statistically reliable alternative to monitoring all
critical planktonic community measures of habitat perturbance
• appropriate for the spatial and temporal scale demanded by the study
objectives
It is recommended that multiple pollution-tolerant and pollution-sensitive indicator
species are selected by clearly defined criteria. Multiple indicator species often
provide a more complete representation of environmental conditions.
B5.4 Variability in measurements caused by field heterogeneity is quantitatively determined
QA/QC by the analysis of replicate field samples. Replicate sampling should be conducted at
Considerations all field stations where measurements are to be used in comparisons. Analysis of
replicate sample data is necessary for assessing the reliability of such comparisons.
Laboratory performance and calibration should be verified at the beginning and
periodically during the time analyses are performed. Commercially available chloro-
phyll is recommended for use in calibration. Chlorophyll quality control samples are
available from EPA's Environmental Monitoring and Support Laboratory in Cincin-
nati, Ohio. Use of blind, split or other control samples can be used to evaluate perfor-
mance. The "Interim Guidance on Quality Assurance/Quality Control (QA/QC) for
the Estuarine Field and Laboratory Methods" (U.S. EPA, 1985) provides a standard
operating procedure for chlorophyll measurements.
-------
B-96
vCi.vXiMk.4M..
Consideration of statistical strategies will mitigate the high costs of collecting and
processing samples. Also see Statistical Design Considerations: Statistical Power
Analysis and Power-Cost Analysis (Appendix B Introduction; Section B.3).
Temporal Stratification of the Data
Temporal stratification of the data should not be attempted until sufficient knowledge
of long-term natural cycles is attained. Initially, simple regression analyses may be
conducted on seasonally stratified data in order to identify monotonic temporal trends.
Further examinations of whether conditions are improving or degrading over time may
be conducted using statistical time series analyses (e;g., temporal autocorrelation,
spectral analyses, etc.)
Information derived from water quality monitoring will be important in interpreting
the results of plankton sampling. For example, phosphate concentrations may indicate
the cause of a phytoplankton "bloom," while measures of dissolved oxygen levels may
describe the consequences of the "bloom" to other living estuarine resources. There-
fore, the selection of water quality and plankton sampling strategies must not be done
independently. These programs should be integrated to the fullest extent possible to
allow correlation of observed responses to changes in water quality parameters. Also,
alterations to the plankton community should be analyzed in relation to other impacts
on biological resources such as food web impacts on fish communities.
B5.5
Statistical Design
Considerations
Plankton monitoring strategies should be able to delineate between natural variability in
plankton stocks and those caused by anthropogenic changes in nutrient concentrations.
Characterization of phytoplankton species abundance, distribution and primary productiv-
ity provide indications of water quality conditions. Monitoring changes in phytoplankton
community composition and densities are critical for the interpretation and evaluation of
long-term trends in water and habitat quality. Further understanding of the causes of
excessive water column and sediment oxygen demand requires tracking of photosynthetic
activity and metabolic rates over time (Chesapeake Executive Council, 1988). Zooplank-
ton abundance and distribution are affected both by changes in phytoplankton and changes
in predator populations. Therefore, population characteristics of this group can indicate
symptoms of water quality problems, fishing pressure and other habitat problems for
predator species (Chesapeake Executive Council, 1988).
B5.6
Use of Data
-------
B-97
B5.7 Rationale
Summary and
Recommendations • Track phytoplankton and herbivore populations if eutrophication is sus-
pected
• Purpose of monitoring is to assess the effectiveness of the CCMP in
mitigating the potential impacts caused by changes in the planktonic
community biomass and structure/function
Monitoring Design Considerations
• Collect both phytoplankton and zooplankton samples at specific depths
throughout the water column
• Phytoplankton samples taken with water bottles and/or pumps can be used
for estimation of chlorophyll concentrations
• Zooplankton sampling methods based on size of target organisms
• Monitoring programs should consider natural and temporal fluctuations in
plankton biomass and species composition
• Taxonomic analysis should include identification and counts of dominant
species
• Other components of the overall monitoring program, including water
quality and fish and shellfish communities, should be integrated with
plankton community data to establish relationships and trends
• Selected biological indices should retain biological meaning, be robust
indicators of estuarine "health", and incorporate species form and function
• Indicator species should possess the following characteristics:
- sensitive to planktonic perturbances of concern
- cost-effective and statistically reliable alternative to measuring all
species in a monitoring program
- statistically reliable indicative measures of habitat perturbance
- appropriate for the spatial and temporal scale determined by the
study objectives
-------
B-98
s- • 4.,.;
" '
• Selection of reference sites is key to the evaluation of environmental
impact due to anthropogenic perturbances; several reference sites may be
required to provide proper control for sampling sites
Existing Analytical Methods
• It is recommended that consistent types of sampling gear, and location and
timing of sample collection be implemented to allow for comparisons
among studies
• Chlorophyll can be estimated using fluorometric or spectrophotometric
methods using a variety of filtration and extraction techniques
• Phytoplankton productivity can be measured using the 14C light-dark bottle
technique or O2 light-dark bottle method
QA/QC Considerations
• Replicate samples should be taken at all field stations where applicable
• Laboratory performance evaluations and calibrations must be done on a
regularly scheduled basis
• Standard chlorophyll samples, obtained from the EPA, should be used for
calibrations
Statistical Design Considerations
• Power analyses may be applied to determine the appropriate number of
sample replicates required to detect a specified difference
• Temporal integration of the data should not be attempted until sufficient
knowledge of long-term natural cycles is attained
Use of Data
• Detect short-and long-term spatial and temporal trends in overall biomass
and productivity
-------
B-99
r" •$•
*"*• ** **
* - -*A
Detect short- and long-term spatial and temporal trends in species abun-
dance, distribution and composition
Examine relationship between water quality conditions and trends in
plankton community characteristics
Examine relationship between plankton community characteristics and
impacts on other living resources (e.g., fish and shellfish communities)
B5.8 Abaychi, J.K., and J.P. Riley. 1979. The determination of phytoplankton pigments
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">»
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i
B6.0 Aquatic Vegetation
Human activities that contribute the greatest direct impacts to vegetation loss and
modification are those that cause physical alterations to the habitat. Physical alter-
ations may cause habitat loss directly or indirectly by modifying natural processes that
significantly affect aquatic vegetation, such as freshwater inflow, hydrology, sedimen-
tation, and sea-level changes.
Marine and estuarine vegetation may be organized into the following groups:
• Emergent vegetated wetlands
- marshes
- mangroves
• Submerged aquatic vegetation (SAV)
- seagrass beds
- other submerged plant communities
• Macroalgal communities
Although all of these habitat types provide essential functions for living marine
resources, the focus of this discussion is primarily on emergent vegetated wetlands due
to the recognized ecological value of these habitats and their rapid rate of deterioration
and loss in many coastal areas.
Emergent Vegetation
Function
TABLE Btf-t LIST OF TERMS
= erect, rooted, herbaceous vegetation
= the physical, chemical, and biological processes or
attributes of a habitat without tegatd to their
importance to society
Submerged Aquatic Vegetation - aquatic plants mat grow \vJiile wholly submerged
and anchored to the substrate
Values
Wetland
« processes'or attributes that^e valuable or tJeaeS-
cial to society
= Ihose areas that areinundated or saturated by
surfacfe or ground wafer at a frequency and
duration sufficient to support, and that under
normal circumstances do support,^ peyal£nc& of
vegetation typically adapted for life in saturated
" soil conditions. Wetlands generally mclucte
- swamps, marshes, bogs, and similar areas. _ ,«,,,
-------
B-1101
Declines in the quality and quantity of marine habitats are thought to reduce produc-
tion of living marine/estuarine resources and diminish other important values of
habitats. Marine and estuarine habitats perform important functions for all living
marine resources. Aquatic vegetation is used by many species for spawning, rearing,
feeding, migration, and shelter from predators. A positive correlation generally exists
between wetland and submerged aquatic vegetation (SAV) acreage and the abundance
of commercially and recreationally important living marine resources (e.g., fish and
crabs). Populations of living marine resources therefore appear to be limited by the
quantity and quality of aquatic vegetation. In factj the condition of wetlands may
prove to be an effective ecological indicator of the condition of selected bird and
mammal populations.
Furthermore, marine and estuarine habitats, in particular vegetated wetlands (tidal
marshes) and submerged aquatic vegetation, perform other valuable functions to man
and the environment. These functions include erosion and flood protection, water
quality control, and provision of habitats for waterfowl and wildlife. Pressures
brought on by population shifts to coastal areas and associated industrial and munici-
pal expansion will continue to jeopardize coastal wetland resources. Monitoring
changes in aquatic vegetation will permit assessment of impacts to the habitat and the
organisms it supports.
B6.1
Rationale
It is essential to understand the effect of monitoring design characteristics on the
results obtained and to standardize them as much as possible to ensure the comparabil-
ity of monitoring efforts throughout the estuary.
Levels of Monitoring Effort
One, or some combination of the following four levels of monitoring effort may be
chosen:
The lowest level of effort would involve periodic measurement of the areal extent of
wetlands based on small scale aerial photography. Aerial photography may also be
used to estimate the extent of submerged aquatic vegetation in low turbidity areas; in
areas of high turbidity, extensive ground truthing would probably be necessary.
The next level of detail would be to map the extent of the dominant plant species.
Many of the dominant species are readily identifiable from aerial photographs. For
B6.2
Monitoring Design
Considerations
-------
B-111
,*,..
example, Spartina alterniflora has stiff upright stalks, while Spartina patens is more
flexible and tends to bend over during low tide.
A third level might include on site measurements relating to functional aspects of the
system. For example, estimates of underground and aerial biomass of dominant plants
may provide information on primary production and health of the system. Kibby et al..
(1980) discuss various techniques for evaluating emergent vegetation. These tech-
niques may be readily modified to assess submerged vegetation.
Level four monitoring would involve integrated studies that examine many functional
aspects of the wetland or submerged aquatic vegetation and the associated animal
community. Techniques such as the Minimal Habitat Matrix, Wetland Evaluation
Technique and Habitat Evaluation Procedure, discussed in Section B6.3, would be
appropriate.
Sampling Period
The aerial coverage and bed density for SAVs vary from year to year due to cata-
strophic storms, exceptionally high precipitation and turbidity, and other poorly
understood natural phenomena. Therefore, short term monitoring will not provide an
accurate assessments of the trend in SA V coverage, but may rather reflect the impacts
of infrequent or periodic events. In addition, wetland losses in acreage occur at low
enough frequencies that yearly measurements are impractical. Furthermore, the types
of wetland losses that are now occurring appear to be moving away from larger
increments to a smaller form of wetland loss that may fall within the margin of error
for current detection limits.
Seasonal/Daily Considerations - In the conduct of habitat functional assessments,
seasonal variations have to be considered. SAV and wetland sampling should be
conducted during the period of peak biomass. Other factors (e.g. tides, weather
conditions, turbidity) combined with a limited growing season also make for a finite
window of opportunity for data capture.
B6.3
Existing Analytical
Methods
Spatial Trends
The most straightforward approach for assessing trends in the aquatic habitat - and the
one most typically used - is to document the loss of habitat acreage (Table B6-2;
France and Hedges, 1989). Spatial data alone are particularly important because of the
-------
B-1121
public concern over declining habitat acreage (both wetlands and submerged aquatic
vegetation) in the nation's estuarine areas. Good recent information on habitat locations
and boundaries form a baseline for monitoring future changes. However, delineating the
extent of habitats alone does not provide a complete measure of the quality of the habitat
in terms of its value for fish, bird and marine mammal populations.
TABLE B6-2. LIST OF ANALYTICAL METHODS
Spatial Trends
Acreage Lo$s
Functional Trends,
Minimum Habitat Matrix
Wetland EvalualtionTechnique
Habitat Evaluation Procedure
Losses of aquatic vegetation acreage in an area of particular concern may be the
compelling reason for the evaluation of this habitat within the estuary. The primary
sources of spatial information are maps, aerial photography, and remote or satellite
imagery. These tools can serve as a simple visual aid for communicating the problem
to the public as well as a main data source for wetland structural and functional
characteristics and assessment of cumulative impacts and trends. Aerial or remote
monitoring can measure changes in acreage, cover types, drainage, configuration and
other spatial parameters related to overall habitat condition and health.
Maps—Maps are the least expensive spatial data source and are also the most simpli-
fied. Generally, maps do not capture all visible characteristics that can be observed
from aerial photographs, and rather selectively portray a subset of what is in an area.
However, maps such as U.S. Geological Survey Topographic Quadrangle maps or
U.S. Fish and Wildlife Service National Wetlands Inventory (NWI) maps can be used
as a base of information from which to compare changes in wetland acreage.
The main benefits of maps for identifying habitats is in their ease of use and low cost.
Their main disadvantage is that habitats are often not clearly or accurately delineated
on maps, and if they are, they are generally not updated frequently enough to provide
timely information on trends. For instance, although the Fish and Wildlife Service has
mapped over 90% of the wetlands in the coastal zone of the lower 48 states, many of
these maps are over 10 years old.
-------
B-113
There have been efforts by some states and the Federal Government to develop high-
resolution automated databases of habitat types through a process of digitizing NWI
maps at a resolution of approximately one acre. The utility of high-resolution and
geographically referenced databases for resource managers has been evaluated by U.S.
Fish and Wildlife Service (Dahl, 1987). Digitizing NWI maps would make updating
information on the status and trends of habitat types easier and more accurate; how-
ever, the progress toward digitizing all coastal wetland maps has been hampered by the
time-consuming and costly nature of the process involved.
Aerial Photography - The most broadly used source of habitat spatial information is
aerial photography. Aerial photographs can be produced at a range of scales and
spatial resolution. Small scales can be used when, for example, very fine detail is
desired for mapping vegetation cover types or complicated drainage patterns. Large
scale coverage can be used when a broad area of coverage is desired. In addition, color
infrared photography can be used to detect differences in soil moisture and subtle
differences in vegetation types accentuating the upland/wetland boundary.
Existing aerial photographs are inexpensive. However, new overflights can be expen-
sive and are often complicated, requiring professional assistance. The advantage of
aerial photographs is that they generally contain more detail than maps of the same
scale or than satellite imagery. When planning overflights for aerial photography, the
following factors must be considered:
tides
weather conditions
• time of day (affects sun angle, atmospheric haze and cloud cover)
• turbidity
Photographs from new overflights can be compared to base maps or older aerial
photographs to record changes over time.
Remote Sensing/Satellite Imagery - Satellite imagery is a useful source of spatial
data when very large study areas are involved and fine detail is not important. Al-
though expensive, when large study areas are involved, satellite imagery may become
more economical than aerial photography. Other than cost, a significant constraint in
the use of satellite imagery is the lack of resolution that could result in missing small
-------
B-1141
•MMim
SK*.
habitats or features; remote sensing provides a coarse estimate of size or boundaries.
However, if the area under investigation is characterized by a very large homogenous
habitat, this constraint may not be as important.
Because images are acquired by scanners and stored in digital form, the spectral
properties, brightness values and other digitally stored data can be manipulated to
enhance or reveal characteristics (such as productivity) not always visible from aerial
photography. However, factors affecting water clarity, such as suspended sediments,
may affect these spectral properties. These factors, and the amount of cloud cover
over the estuary, should be considered when appraising the use of remote sensing.
Ground-truthing of satellite images is highly recommended in order that accurate
interpretations of the images may be achieved.
When analyzing remotely sensed images, the availability and effectiveness of appro-
priate algorithms may be limited (Platt and Sathyendranath, 1988). Interpretation,
manipulation, and analyses of remotely sensed images requires persons with special-
ized skills handling image analyzing software and hardware. Furthermore, some
habitat assessment methods require a high degree of advanced understanding of the
science involved in the assessment, such as plant taxonomy or hydrology. The selec-
tion of the assessment method will therefore depend on the level of specificity required
to conduct the assessment and the available technical expertise.
Transects and Quadrats-Physical and biological attributes of wetland vegetation
may be taken at regular intervals along a transect: point intercept method (SCS, 1976).
Critical biological characteristics include species name, plant and mulch biomass, and
foliar and basal cover. Physical characteristics (e.g., salinity, water depth, flow rates)
should also be collected to assist in the interpretation of the biological data.
Plants may be harvested from standard-sized quadrats (e.g., 1 m2) along a transect
(SCS, 1976). Conversion factors are available to convert plant species data from
biomass per plot to more common measures of productivity (e.g., pounds/acre or
kilograms/hectare). Measures should specify green weight or air-dry weight; factors
for converting between green weight to air-dry weight exist.
Measures of cover are fundamental to managing living resources of wetlands. Basal
cover at the erosion interface provides information concerning retention of detrital
material and potential for erosion. Basal cover complements measures of productivity
by providing information concerning the stability of the habitat. For example, al-
though a plant community may be highly productive, it may be susceptible to erosion
-------
B-115
-\
if there exists little basal cover at the erosion interface. This information may be used
to guide implementation of erosion control measures.
The U.S. Department of Agriculture (USDA) has developed the Range Data System
(RDS), a nationwide database system for grass-based ecosystems. RDS is capable of
storing, retrieving, and analyzing data on wetland plant productivity and species
composition. RDS also provides a remarks field where data such as salinity and water
depth may be recorded.
Functional Trends
The purpose of assessing habitat functional trends is to compare the relative value of
one habitat to another or to assess changes in values of the same habitat over time.
Detailed, site-specific, functional assessment studies can be expensive, time-consum-
ing, and often impractical when time or budgetary constraints exist. Although more
expensive, correlating functional habitat losses to acreage loss is a more meaningful
measure of the condition of estuarine resources than acreage loss alone.
Where time and budget allow, implementation of detailed, carefully designed monitor-
ing programs or use of quantitative computer models usually give superior-quality
results (Table B6-2). All functional assessment methods, however, are limited by the
level of understanding of the actual processes taking place in habitats, which vary with
habitat type, region of the country, and physical, chemical and biological conditions.
Functional assessments usually describe trends in water quality, hydrology, and biota ;
that are potentially attributable to habitat loss and impacts.
Standardized, relatively rapid functional evaluation procedures generally provide
stronger replicability and technical comprehension. Two of the most frequently
standardized evaluation techniques are the Wetland Evaluation Technique (WET),
originally developed for the Federal Highway Administration and later revised for the
U.S. Army Corps of Engineers (Adamus et al., 1987) and the Habitat Evaluation
Procedure (HEP), developed by the U.S. Fish and Wildlife Service (U.S. FWS, 1980).
Many of the more recently developed habitat evaluation methods are modified ver-
sions of these techniques.
Minimum Habitat Matrix - As part of the Chesapeake Bay Program, minimum
habitat guidelines for various species are developed with the ultimate goal of reestab-
lishing a balanced ecosystem. This method is designed to provide information on the
minimum habitat quality needed by a target species and identifies those factors (both
-------
B-116
environmental and ecological) required for the species. This information is formatted
into a habitat requirement matrix that defines the habitat parameters needed for suc-
cessful reproduction and survival of the indicated species.
The matrices can indicate the vital environmental parameters that should be monitored
and thus facilitate the establishment of monitoring programs. This process is used to
estimate the feasibility, benefits and potential costs of maintaining and protecting an
cstuarine environment suitable for the successful reproduction and survival of aquatic
vegetation (Chesapeake Executive Council, 1988).
One potential problem with this method is that only the target species and those
organisms that the target species depends on for food are tracked, with the intention of
maintaining habitat quality for both groups. This method may not completely recog-
nize the complex species interdependence within estuarine environments.
Wetland Evaluation Technique (WET) - The Wetland Evaluation Technique
outlines the procedure for conducting an assessment of the following wetland func-
tions and values:
Ground Water Recharge
Ground Water Discharge
Flood Flow Alteration
Sediment Stabilization
Sediment/Toxicant Retention
Nutrient Removal/Transformation
Production Export
Wildlife Diversity/Abundance
Aquatic Diversity/Abundance
Recreation
Uniqueness/Heritage
WET assesses the suitability of wetland habitat for 14 waterfowl species groups, 4
freshwater fish species groups, 120 species of wetland-dependent birds, 133 species of
saltwater fish and invertebrates, and 90 species of freshwater fish. WET does not
evaluate any other important wildlife resources (e.g., game and furbearing mammals).
Other evaluation methods must be used to evaluate these other wildlife resources
(Adamus et al, 1987).
WET assesses functions and values by characterizing a wetland in terms of its physi-
cal, chemical, and biological processes and attributes (Adamus et al., 1987). This
characterization is accomplished by identifying threshold values for predictors. Pre-
dictors are simple, or integrated, variables that directly, or indirectly, measure the
physical, chemical, and biological processes or attributes of a wetland and its sur-
roundings.
-------
B-117
WET evaluates functions and values in terms of social significance, effectiveness, and
opportunity (Adamus et al., 1987). Social significance assesses the value of a wetland
to society due to its special designations, potential economic value, and strategic
location. Effectiveness assesses the capability of a wetland to perform a function due
to its physical, chemical or biological characteristics. Opportunity assesses the oppor-
tunity of a wetland to perform a function to its level of capability.
Habitat Evaluation Procedure (HEP) - The Habitat Evaluation Procedure is a
method that can be used to document the quality and quantity of available habitat for
selected wildlife species. HEP provides information for two general types of wildlife
habitat comparisons: (1) the relative value of different areas at the same point in time;
and (2) the relative value of the same area at various points in time. By combining the
two types of comparisons, the impacts on, or improvement in habitat quality as a result
of proposed or anticipated land and water changes on wildlife habitat can be quantified
(Lonard and Clairain, 1986).
In the evaluation procedure, the habitat quality for selected species is documented
based on an evaluation of the ability of key habitat components to supply the life
requisites of the selected species. The evaluation involves using the same key habitat
components to compare existing habitat conditions and the optimum conditions for the
species of interest (U.S. FWS, 1980).
There are a number of limitations to using a habitat approach, such as HEP, in an
evaluation system as was pointed out by the U.S. Fish and Wildlife Service (1980).
Using habitat quality as an evaluation standard limits the application of the methodol-
ogy to those situations in which measurable and predictable habitat changes are an
important variable. It also forces a long-term "averaging" type of analysis. There is
no assurance that populations will exist at the potential levels predicted by habitat
analysis, as the analysis may not include all of the environmental or behavioral vari-
ables that may limit populations below the predicted habitat potential. In addition,
socio-economic or political constraints may prevent the actual growth of certain
populations to these levels (U.S. FWS, 1980).
B6.4
QA/QC
Considerations
Whichever monitoring method is selected, the researcher should ensure that it has a
high degree of replicability. All functional assessment methods contain assumptions
that impact the results of the assessment. Some assessment methods also require
excessive judgement on the part of the researcher and as a result, different users may
arrive at different results. In selecting a method, the researcher should first determine
whether assumptions are based on scientific knowledge or on the opinion of the author.
-------
B-1181
Before applying a method extensively within a region, the researcher should field test
the method by comparing its ratings to those determined independently by long-term
research studies of representative habitat sites. Ground-truthing remotely sensed and
aerial images is highly recommended in order to ensure accurate interpretation of the
images.
In designing an aquatic vegetation monitoring program the researcher must first
determine the resources available. This will determine the extent of the monitoring
program and the level of detail it can provide. If resources are limited, the extent of
the monitoring program may only involve an assessment of the aereal extent of
wetlands. If more resources are available and it is decided to undertake a functional
assessment, the researcher must decide which functions are of greatest interest in the
study area or which will provide the most information concerning the habitat attributes
perceived to be of greatest value to the area.
Due to time and budgetary constraints, it may not be possible to evaluate all of the
habitats within the region of concern (e.g., an estuary). In such cases, the researcher
may choose to evaluate only a select few habitats. These may be those with poten-
tially the greatest ecological value, those undergoing remediation, or those which are
undergoing significant loss or modification.
Most habitat evaluation techniques involve some form of ranking, either quantitative
or qualitative. The purpose of the monitoring in a estuary program is to periodically
compare baseline conditions to conditions after the management plan has been imple-
mented. In terms of habitat monitoring this means comparing the baseline acreage or
functional rating of the habitat to conditions some time after the management plan has
been implemented. Therefore, replicability of results is imperative.
See also Statistical Design Consideration: Statistical Power and Power-Cost Analysis
(Section B.3).
B6.5
Statistical Design
Considerations
The results of aquatic vegetation evaluation will track trends in habitat quality and
quantity so that they may be projected into the future. Additionally, changes in land
use or other anthropogenic activities over time can be measured in terms of their
impact upon aquatic habitats. It is a primary concern to accumulate an adequate set of
data to establish a baseline so that any changes in habitat quantity and quality that are
derived from mitigation, regulation or an estuary management program can be as-
sessed. Historical data can chart changes in land use leading up to the present state.
B6.6
Use of Data
-------
The tracking of habitats may be an effective method of monitoring estuarine bird and
mammal populations. These organisms are highly mobile and their presence or
absence at particular monitoring stations may not be significant - individuals may be
situated in another similar location. However, monitoring habitats, which these organ-
isms require (e.g., to nest, breed, feed, use for shelter from predators, etc.) may be an
effective means of monitoring the threat to their well-being in the estuary. Loss of
biologically critical habitats will indicate the potential decline or loss of estuarine bird
and mammal populations.
Although single coastal development projects that result in modification or loss of
estuarine vegetation may not have serious immediate environmental repercussions
beyond the point of initial impact, many such projects in succession over time and over
a broad geographic area may have far-reaching ecological consequences (Day et al.,
1989). Without reasonably accurate information on long-term and cumulative impacts
of human activities on marine and estuarine habitats, good decisions on permitting,
mitigation, or management of fisheries and wildlife will be difficult.
B6.7
Summary and
Recommendations
Rationale
Monitoring habitat quantity and quality will provide information to assess impacts on
living resources and to gauge the effectiveness of mitigation, regulatory and manage-
ment programs.
Monitoring Design Considerations
• Possible resilience of habitat functions should be considered; need to be
able to assess long-term and cumulative impacts
• Seasonal and daily variations may have an impact on habitat functions and
the ability to identify habitat boundaries and thus affect evaluation results
• Some methods require a high degree of technical understanding while other
methods can be easily understood and implemented with more limited -
technical understanding
-------
B-120!
Existing Analytical Methods
• Analyses of spatial trends is the most straightforward approach for assess-
ing trends in the aquatic habitat. Typically, documentation of habitat
acreage loss is assessed.
• Maps have the following properties
- least expensive
- easy to use
- most simplified: do not capture all features
- habitats not always clearly or accurately delineated
- often not updated on regular basis
• Aerial photography has the following properties
- range of scales and spatial resolution
- color infrared photography to detect differences in soil moisture
and vegetation types
- relatively inexpensive
- contain more detail than maps or satellite imagery
• Satellite imagery has the following properties
- useful when large study areas involved and great detail not
important
- spectral properties, brightness values and other digitally stored
data can be manipulated to enhance characteristics
- expensive
- lack of resolution found in aerial photographs
• Functional analysis considerations
- provide more detailed information concerning the quality of
habitat
- can be expensive and time consuming
- limited by the level of understanding of actual processes
- WET and HEP two of the most frequently used standardized
evaluation techniques
QAIQC Considerations
• Need high degree of replicability within a method
-------
B-121
• Method should be field tested before being applied extensively
Statistical Design Considerations
• Available resources
• Determine which functional assessments will provide information concern-
ing attributes of greatest value to the area
• Determine area! extent of monitoring program
- entire watershed
- select habitats with greatest potential value or undergoing signifi-
cant loss or modification
Use of Data
• Project future trends in habitat quality and quantity
• Evaluate severity of habitat impacts from changes in land use or other
anthropogenic activity over time
• Assess improvements in habitat quality and quantity resulting from mitiga-
tion, regulation, and estuary management programs
• Ecological indicator of potential decline or loss of estuarine bird and
mammal populations
B6.8 Ackleson, S.G. and V. Klemas. 1987. Remote sensing of submerged aquatic vegeta-
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pollutant reduction in buffer zones adjacent to land treatment sites. Trans. ASCE
24:430-435.
Phillips, R.C. and C.P. McRoy. 1990. Seagrass Research Methods. UNESCO,
Monographs and Oceanographic Methodology, No. 9.
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B-125
Platt, T. and S. Sathyendranath. 1988. Oceanic primary production: Estimation by
remote sensing at local and regional scales. Sci. 241:1613-1619.
Reppert, R.T., Siglero, W., Stakhiv, E., Messman, L., and Meyers, C. 1979. Wetland
values: Concepts and methods for wetlands evaluations. IWR Research Report
79-R-l, U.S. Army Engineer Institute for Water Resources, Fort Belvoir, Virginia.
Rose, P.W., and P.C. Rosendahl. 1983. Classification of Landsat data for hydrologic
application, Everglades National Park. Photo. Eng. Rem. Sen. 49:505-511.
SCS. 1976. National Range Handbook. U.S. Department of Agriculture, Soil Conser-
vation Service.
SCS Engineers. 1979. Analysis of selected functional characteristics of wetlands.
Contract No. DACW73-78-R-0017, Reston, VA.
Shears, J.R. 1989. The application of airborne remote sensing to the monitoring of
coastal saltmarshes. In: Remote Sensing for Operational Applications. Technical
Contents of the 15th Annual Conference of the Remote Sensing Society, University of
Bristol, September 13-15,1989. Barrett, E.G., and K.A. Brown, eds. pp. 371-379.
Shudiiner, P.W., Cope, C.F., and Newton, R.B. 1979. Ecological effects on highway
fills of wetlands. Research Report. National Cooperative.
Simon, B.D., LJ. Stoerzer, and R.W. Watson. 1987. Evaluating the functional
significance of wetlands for flood storage. Wisconsin Dept. Natur. Resour., Madison,
Wisconsin.
Solomon, R.D., Colbert, B.K., Hanses, W.J., Richardson, S.E., Ganter, L.W. and E.G.
Vlachos. 1977. Water Resources Assessment Methodology (WRAM) - Impact
Assessment and Alternative Evaluation. Technical Report Y-77-1, Environmental.
Effects Laboratory, U.S. Army Engineer Waterways Experiment Station, CE,
Vicksburg, MS.
State of Maryland Department of Natural Resources. Undated. Environmental
Evaluation of Coastal Wetlands (Draft). Tidal Wetlands Study, pp. 181-208.
State University of New York at Syracuse (SUNY). 1987. Wetlands evaluation
system for Onondaga County, New York State. Draft. 93 pp.
-------
B-1261
•ft
ft •iXww.'SWF tf
^H \
-Xviv >
Stevenson, J.C. and N. Confer. 1978. Summary of Available Information on Chesa-
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co-sponsored by Maryland Dept. of Natural resources and U.S. Environmental Protec-
tion Agency. 335 pp.
Sweet, B.C., J.E. Poppleton, A.G. Shuey, and T.O. Peeples. 1980. Vegetation of
central Florida's east coast use the distribution of 6 vegetational complexes of Merritt
Island and Cape Canaveral peninsula. Rem. Sen. Env. 9:93-108.
Ulanowicz, R.E., and D. Baird. 1986. A Network Analysis of the Chesapeake Bay
Ecosystem, Maryland department of Natural Resources Tidewater Administration.
Ref. No. [UMCEES] CBL 86-79.
U.S. Army Engineer Division, Lower Mississippi Valley. 1980. A habitat evaluation
system for water resources planning. US Army Corps of Engineers, Lower Missis-
sippi Valley Division, Vicksburg, MS.
U.S. Army Engineer Division, New England. 1972. Charles River: Main Report and
Attachments. Waltham, Massachusetts.
U.S. Department of Agriculture. 1978. Wetlands evaluations criteria-water and
related land resources of the Coastal Regions, Massachusetts. Soil Conservation
Service, Amherst, Massachusetts.
U.S. EPA. 1987. Report of the workshop on habitat requirements for the Chesapeake
Bay living resources. Prepared by Eastern Research Group, Inc.
U.S. EPA. 1988. Use of Geographic Information Systems for Wetlands Protection.
Washington, D.C.: Office of Wetlands Protection, US EPA.
U.S. Fish and Wildlife Service (U.S. FWS). 1980. Habitat Evaluation Procedures
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U.S. Fish and Wildlife Service. 1982. Standards for the development of habitat
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Virginia Institute of Marine Science. Undated. Evaluations of Virginia Wetland
(Mimeographed). Glouchester Point, VA.
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B-127
B7.0 Benthic Infauna Community Structure
B7.1 The objective of monitoring the benthic infauna is to detect and describe spatial and
Rationale temporal changes in the structure and function of benthic communities. These results
can be used to assess the condition - the "health" - of benthic habitats, to monitor rates
of recovery following environmental interventions, and, since the benthic infauna
represent a significant food source for many estuarine organisms, to provide an early
warning of potential impacts to the estuarine ecosystem.
The benthic habitat may broadly be divided into hard- and soft-bottom habitats. Since
the predominant habitat in estuaries is soft-bottom, assessment of soft-bottom commu-
nity structure will be the emphasis of this section.
Monitoring the abundance of individual organisms in the benthic infauna is a widely-
accepted means of measuring the condition of the benthic habitat (Bilyard, 1987).
Benthic infaunal organisms are exceptional indicators of benthic conditions since:
• they are generally sedentary - observed effects are in response to local
environmental conditions
• they are sensitive to habitat disturbance - communities undergo dramatic
changes in species composition and abundance in response to environmen-
tal perturbations
• they often mediate the transfer of nutrients and toxic substances in the
ecosystem - via bioturbation and as important prey organisms
Furthermore, monitoring benthic infauna is one of a handful of methods which pro-
vides in situ measures of biotic health and habitat quality. The assessment of benthic
infaunal community structure is a powerful tool in the evaluation of spatial and
temporal effects of anthropogenic and natural disturbances.
B7.2 The level of effort required to assess benthic community structure is relatively high. A
Monitoring Design field survey is required to collect organisms; sorting, identifying, and enumerating
Considerations specimens requires labor-intensive and generally expensive processes. Evaluations of
benthic community structure and function may also be costly and time-consuming.
The results of benthic monitoring programs can vary substantially depending on the
objectives and corresponding design specifications. The characteristics that are
primarily responsible for the variability in the results are:
-------
B-1281
I Mill "
I ipi M'llil I until
ii x t*Mf*. "VW%&
'*~»Ss
4 A?>
• type of sampling gear
• sample sorting protocols
• level of taxonomy
• location and timing of sample collection
It is essential to understand the effect of these design characteristics on the results and
to standardize them as much as possible to ensure the comparability of monitoring
efforts throughout the estuary.
Analyses of power-cost efficiencies are essential in selecting the appropriate sampling
gear and sample processing protocols. Ferraro et al. (1989) provide an example of
power-cost analyses.
Sampling Devices
Sample collection protocols influence all subsequent laboratory and data analysis; it is
key that benthic samples be collected using acceptable and standardized techniques.
Several types of devices can be used to collect benthic macroinvertebrate samples:
trawls, dredges, grabs, and box corers (Mclntyre et al, 1984). Many of these devices
sample the benthos in a unique manner. Accordingly, conducting comparisons among
data collected using different devices is inadvisable. It is recommended that the same
area and volume of the sediment be sampled since different species of benthic
macroinvertebrates exhibit different horizontal and vertical distributions (Elliot, 1971).
Furthermore, collection of sediments and benthic organisms should be done concur-
rently in order to mitigate the costs of field sampling and to permit sound correlation
and multivariate analyses (see Sections B.3 and B.4). Therefore, it is recommended
that the sampling device also be suitable for sampling the sediment (Table B7-1).
Collection of an acceptable sediment sample for infaunal analyses requires that the
sampler
• create a minimal pressure wave when descending
• form a leakproof seal when the sediment sample is taken
• prevent winnowing and excessive sample disturbance when ascending
-------
B-129
TABLE-B7-L SUMMARY OF BOTTOM SAMPLING EQUIPMENT8
Device
Use
Advantages
1 % Disadvantages
Fluorocarbon plastic of Shallow wadeable waters or
Glass Tube deep waters if SCUBA
'' available. Softorsemi-
-- consolidated deposits*
HandCorer^ith
-removable Fluorocar-
bon plastic or glass
liners: , , -
Boxcorer
Same as above except more
consolidated sediments Can
"Same, as above,
Gravity carers, that is, ^ " Semi-consolidated sedi-
PhlegerCorer ' ments,
.Young Grab {fluorocar- Lakes and marine areas.
bon plastic- or kynar-
lined modified &l m2
van. Veen)
Ekman or BOX Dredge Soft to semi-soft sediments.
- „, „,,/ Can be used from boat,
v % " bridge, or pier in waters of
Various depths.
P0KAB. Grab Sampler
on sand, silt, or clay.
iTeserves layering and
permits historical study of
sedimentfdeposition. Rapid-
samples Immediately ready
for laboratory shipment
Minimal risk of contamina-
tion.
Handles provide for greater
ease of substrate penetration.
Above advantages*
Collection of large undis-
turbed sample allowing for
subsampling,
-";.,
Low risk of sample contam>
nation^ Maintains sediment
integrity relatively well.
Eliminates metal contamina-
tion. Deduced pressure
wave.
Obtains a larger sample than
coring tubes. Can be
subsampled through box Itd^
Most universal grab sampler*
Adequate Qmnostsub-
strafes. Large sample
obtained intact, permitting
subsampling,
Small Sample size requires
repetitive sampling. ,,..
Careful handling necessary to
prevent spillage, Requires
removal of liners before
repetitive sampling, Slight
risk of metal contamination.
from barrel and corecuttfer^
Hard to handle;
Careful liandling-necessary to%
avoid sediment/spillage.
Small sample, respires
repetitive operation and
removal of liners, Time
consuming..
Expensive. Requires winch,
Possible incomplete j£w
closure and sample Joss.
Possible shock wave which
may disturb the fines. Metal
construction may introduce
contaminants. Possible loss
of "fines'* on retrieval, - Nv"~c
Shock wave from descent, ^,
nuay disturb "fines". Possible
incomplete closure of jaws „
results in sample loss.
Possible contamination ftom.
metal frame construction.
Sample imjst be further „
prepared for analysis. -
-------
B-1301
TABLE B7-1 SUMMARY OF BOTTOM SAMPLING EQUIPMENT8
s Xv1. fftftf
, ^ (continued) , ,,,, ,, ,
Device
"Tfea
Advantages
Disadvantages
BMH-53 Piston Corer Wat9rs'6f4-^ftdeepwhen piston provides fcr greater
used with extension xodu sample retendon.
Soft to semi-consolidated
deposits, ^v, ^ _ "-
Van Veen
BMH-60
from a fixe$ platform
s
tlseful ,on ^nd> sijt» or play. Adequate on m(»t sub»
, -%' " strates, large sample
- obtained intact, permitting
< x>" -s " subsampling.
Streamlined configuration
allows sampling where other
devices Could not achieve
proper orientation,
..Carge samptej can penetrate
most substrates.
Sample bucket may be
opened to permit
subsampling. Retains fine s
v. grained sediments effeo '
lively*
Designed for sampling hatd
substrates.
Inexpensive/easy to handle.
Used primarily telnariite
^waters
^ s
Peterson Grab Sampler Useful^n'mpsVsubstrates,
%
Shipelc Grab Sampler
Orange-Peel Grab
Smith-MclnfyrftGrab
Scoops, Drag Buckets
Cotes must.be extruded o»
Site to other containers-metal
barrels introduce risk of ss s
metal contamination.
Shock wave from desdent
.may disturb "fines*': Possible
Incomplete closure ojf Jaws
results in sample loss.
Possible contamination from
metal frame, construction*
"• Sample must be furtfiet
prepared for analysis.
s Possible coMtamlnation from
" metal construction. " ,
SubsampHngdifRculfc Hot
eirectiye for sampling fine
sediments,
Heavy, may require winch.
No cover lid to permit
Subsampling, All Other
disadvantages of Ekman and
Ponar*
Possible contamination from
- metal construction. Heavy,
may require winch.
Useful on most substrates,
Various environments
depending onxdepth and, s
' * "
Loss of fines,
require winch, Possible
' metal contamination.
toss of fines on retrieval
through water column.
* Comments represent subjectivfe euations, ^,,"" •. -
SOURCE: American Societytor Testing a^Materiafe, 1991
-------
B-131
• allow easy access to the sample surface in order that undisturbed
subsamples may be taken
Penetration well below the desired sampling depth is preferred to prevent sample
disturbance as the device closes. It is optimal to use a sampler that has the means of
weight adjustment in order that penetration depths may be modified.
Trawls and Dredges - Trawls and dredges generally collect organisms over a variable
and relatively large area (Fredette et al., 1989; U.S. EPA, 1986-1991). Their design ;
and use precludes the collection of infaunal organisms at sediment depths greater than
a few centimeters.
The data collected using these devices are qualitative since consistent and reproducible
sampling of a constant area of the bottom is not possible. However, these devices are
suitable for reconnaissance of potential sampling sites. Data collected using trawls or
dredges may be used to generate a list of organisms present in the area. A list of
species may be used to select sampling locations for a monitoring program.
Grab Samplers - Grab samplers are capable of consistent sampling of bottom habi-
tats; depending on the size of the device, areas 0.02 to 0.5 m2 and depths ranging from
5 to 15 cm may be sampled. Limitations of grab samplers include:
• variability among samples in penetration depth depending on sediment
properties
• oblique angles of penetration which result in varying penetration depths
within a sample
• the sample is inevitably folded resulting in the loss of information concern-
ing the vertical structure of benthic communities in the sediments
However, the careful use of these devices will provide quantitative data. Grab sam-
plers are the tools of choice for a number of estuarine and marine monitoring programs
due to their ability to provide quantitative data at a relatively low cost (Fredette et al.,
1989; U.S. EPA, 1986-1991).
Core Samplers- Core samplers utilize a surrounding frame to ensure vertical entry;
vertical sectioning of the sample is possible (U.S. EPA, 1986-1991). These devices
are capable of maximum penetration depths of 15 cm and may collect volumes 5 to 10
times that of grab samplers. Limitations of box corers include:
-------
B-1321
• its large size and weight require the use of cranes or winches and a large
vessel for deployment
• higher construction expenses
• lack of calibration studies to permit comparisons to grab samples
The Hessler-Sandia box corer uses dividers to section the core into subsections,
facilitating subsampling of the core. Box corers are recognized as the tools of choice
for maximum accuracy and precision when sampling soft-bottom habitats.
Suction Samplers -In situ suction devices use the flow of water to collect sediments
and bcnlhic organisms (Eleftheriou and Holme, 1984). An open-ended suction tube
draws sediments and organisms into the flow of water, organisms are trapped in a
sieve or mesh bag at the end of the tube. Limitations of suction devices include:
• little control of the volume of sediment collected
• collection of animals from surrounding sediments, inflating infauna
abundances
• organisms are typically turbulently abraded during collection, making
identifications even more difficult
Flcischack et al. (1985) propose modifications to suction samplers that overcome some
of these limitations. Some samplers can be operated remotely; more often they are
operated by divers. Any diver-operated collection procedures will be restricted to
SCUBA depths and conditions permitting safe diving (e.g., relatively calm waters,
reasonable visibility) thereby limiting their use in monitoring programs.
Sediment Profiling Camera - The sediment profiling camera is a unique tool which
allows vertical, in situ imaging of the water-sediment interface. Photographs of
biological activity may be made at a maximum depth of 18 cm, however, penetration
depth of the viewing prism may be limited due to the physical characteristics of the
sediment (i.e., penetration depths are greater in silt than sand).
The sediment profiling camera system can give measures of organism tube density,
thickness of the pelleted layer, and infauna present in the image area (Rhoads and
Gcrmano, 1982). Although this tool provides qualitative information concerning the
-------
B-133
v
benthos activity, the sediment profiling camera system cannot provide quantitative
information on species diversity, abundance, and biomass. Information concerning
species composition cannot be gained using this tool. Consequently, characterizations
of the benthic community structure cannot be made from the data taken from these
images.
The use of the sediment profiling camera is not recommended for the collection of
benthic community structure data. However, the sediment profile camera proves to be
effective as a reconnaissance tool. Delineation of physically similar sampling sites
may be determined through the use of this tool, aiding in the selection of sampling
stations. Ground-truthing of these images by means of laboratory analyses of collected
material is highly recommended.
Recommendations - If quantitative analyses of benthic community structure are
required, a grab or core sampler is recommended. Either of these sampling devices
permit adequate sampling of both sediment and benthic infaunal communities with the
one sampling device.
Area Sampled
Different species of benthic macroinvertebrates have different scales of horizontal and
vertical spatial distribution (Elliot, 1971; Livingston etal, 1976; Downing, 1979).
Costs of laboratory analyses of the sample increase with increased sample unit size.
Analyses of spatial and temporal scale, statistical power, as well as costs, will assist in
determining optimal sample unit size. It is highly recommended that a standard
sample unit size (same surface area and depth) be analyzed in order to ensure data
comparability.
Sorting Protocols
There are several options which may be considered when designing sorting protocols.
In order to assure comparability among studies, a consistent set of sorting procedures
is highly recommended.
Sieving: Mesh Size and Location - The use of different sieve mesh sizes limits the
comparability of results among estuarine monitoring studies (Reish, 1959; Rees,
1984). The major advantage of using a smaller mesh size is the retention of both
juveniles and adults organisms as well as large- and small-bodied taxa. The major
disadvantage is the concomitant increased cost of sample processing. For example,
-------
B-1341
using a 0.5 mm mesh over a 1.0 mm mesh could increase retention of total
macrofaunal organisms by 130-180 percent; however, increased costs for processing
the samples may be as much as 200 percent (U.S. EPA, 1986-1991).
It is recommended that a standard mesh size be selected for all monitoring studies
within the estuary. In Puget Sound, the 1.0 mm mesh size is commonly used in impact
assessments and the 0.5 mm mesh size is sometimes used in recruitment studies. The
issue of data comparability was resolved by recommending sequential screening and
analysis procedures (Becker and Armstrong, 1988). By screening samples through
both the 1.0 and 0.5 mm sieves, the 1.0 mm fraction could be analyzed separately and
the data compared with other impact studies throughout the estuary. The data for the
0.5 mm fraction of the samples could then be combined to yield information on the
complete sample for comparison with other recruitment data.
Sieving can be conducted either aboard the survey vessel or onshore after the cruise is
completed. The major difference is that aboard the vessel, sieving occurs prior to
fixation; onshore, sieving occurs after fixation. Fixation of organisms in the sediment
before being sieved has the problem of the timely distribution of adequate concentra-
tions of the fixative to all the organisms in the sample. Sieving characteristics of
organisms can be greatly modified by decomposition and deterioration of body parts.
When a large number of samples are to be collected during a cruise, field sieving
results in a significant reduction in the volume of material stored and later transported
to the laboratory.
Use of Relaxants - The use of relaxants prior to sieving and fixation reduces the
tendency of organisms to distort their shape or autotomize, and consequently facilitates
taxonomic identifications and morphometric measurements (U.S. EPA, 1986-1991).
Complete organisms, having a "natural" appearance, are easier to identify to lower
taxonomic groups than are distorted or fragmented specimens.
The magnitude to which relaxants can influence taxonomic identification, thereby
limiting comparisons among monitoring studies, is unknown. It is not recommended
that comparisons be conducted between studies in which relaxants were used and those
in which they were not used.
Use of Stains - Vital stains—primarily rose bengal — are added to samples to
facilitate sorting. The stain colors most infauna, enhancing their contrast with back-
ground debris. Not all taxa stain adequately and some taxonomists have found that
staining interferes with the identification of certain taxa. Although staining may aid
-------
f **
fv-,
B-135
X
"***
-•"• A
the sorting process, a proper quality control program should ensure that sorting effi-
ciency is maintained whether or not a stain is used.
Level of Taxonomy
The necessity to identify organisms to the level of species has recently been questioned
(Warwick, 1986). The liabilities of identifying organisms to the species level include:
• inaccurate and imprecise identifications (Ellis, 1985) • '•
• identifications are time-consuming and costly; costs can be reduced as
much as 30-50 percent by limiting identification of samples to higher
taxonomic levels (U.S. EPA, 1986-1991)
Identifications to higher taxonomic groups can provide gross characterizations of
benthic infaunal assemblages and may be sufficient to meet program objectives
concerning detection of community responses to anthropogenic disturbance. Warwick
(1986) contends that detection of anthropogenic disturbances would be facilitated
since these perturbations impact community structure at taxonomic levels higher than
the species level. Natural disturbances, which generally affect infauna community
structure by species replacement, would have little influence on analyses at higher
taxonomic levels.
However, identification to higher taxonomic levels may sacrifice the potential wealth
of information available using species-level identification, such as analyses of popula-
tion dynamics and productivity. Furthermore, it is generally regarded that the species
is the taxon most responsive to environmental stress; individual species will tend to be
replaced by another species within the same genus before the genus is replaced (U.S.
EPA, 1986-1991).
The level of taxonomic identification will depend on monitoring program objectives,
sample size, study sites, and analytical measures. Data based on lower taxonomic
levels can be grouped for future comparisons with higher level taxa. It is strongly
recommended that all samples identified only to higher taxonomic levels be properly
archived, since comparisons of lower taxonomic data may be required later.
Selection of Sampling Period
Benthic infauna assemblages are dynamic; the most common temporal patterns
observed in benthic communities are those associated with seasonal changes. Seasonal
-------
B-1361
variation in benthic assemblages may be due to changes in physical, chemical, and/or
biological parameters: (e.g., temperature, light transmissivity, dissolved oxygen,
predation, recruitment).
Given the seasonal variation characteristic of benthic assemblages in general, it is
recommended that direct comparisons between samples collected during different
seasons be avoided. Studies investigating interannual variation in the characteristics of
benthic communities should be conducted during the same season (preferably the same
month) each year.
There are a variety of approaches to assess the effects of anthropogenic perturbance on
bcnthic communities of the estuary. Many assessment approaches are derived from
modifications of Connell's (1978) Intermediate Disturbance Hypothesis. Some
researchers have proposed that selected community structure measures — e.g., species
composition, abundance, etc. — are predictable along a gradient of environmental
disturbance (Pearson and Rosenberg, 1978; Gray and Mirza, 1979; Warwick, 1986).
A corollary to this supposition is that indicator species, whose responses would
epitomize community responses to habitat perturbations may be identified.
These assessment approaches may be grouped into three categories:
• biological indices
• indicator species
• multivariate analyses
However, there has been little consensus among biologists regarding the suitability of
various techniques for describing community characteristics and/or for assessing
cstuarine impacts. A critical evaluation of the use of biological indices to detect
environmental change is presented in an EPA Technical Support Document (U.S.
EPA, 1985; Table B7-2). In addition to measures of changes in the abundances of
pollution sensitive, pollution tolerant, and opportunistic species, the indices shown in
Table B7-2 are evaluated on the basis of the following criteria:
• biological meaning
• ease of interpretation
B7.3
Existing Analytical
Methods
-------
B-137
TABLE B7-2. BIOLOGICAL INDICES
Index/Method
Biological
Characteristic Measured
Recommended
W Monitoring*
- Biological integrity
Bray-Curtis
Dominance1*
Maunal index
No. individuals
No< species
Community structure
Dissimilarity
Community structure'
Community structure
Total aljundance
Total taxa
Opportunistic and Community structure
pollution tolerant species
Pollution-sensitive
species
Biomass
Margalefs SR "
Pielou's J
Shannon-Wiener H1
Community structure
Standing crop
Diversity
Evenness
Diversity
B
P,B,F
Be
P.B.P1
P.B,?3
al> fclatokton)* B (benthos), aodF (fishes) indicate thos& biological groups to which
^ a given index may be applied.
^Defined as the minimum number of species required to account for 75 percent of -
the individuals in asample (see Swartz et <#„ 1985),
cWhere developed, "
dMay be used together as additional indices of community structure, ,.. ,
-------
B-138 ]
• sensitivity to community changes due to anthropogenic sources
The results of these evaluations and additional information on other analytical methods
are summarized below.
Biological Indices
The numbers of individuals and the numbers of species have been found to be good
indicators of anthropogenic disturbance, as well as of other environmental stresses
(Table B7-2; Pearson and Rosenberg, 1978; Warwick, 1986). Furthermore, these
simple biological indices are less ambiguous and are often as informative as diversity
indices (U.S. EPA, 1985; Green, 1979; Hurlbert, 1971). Measures of biomass have
inherent problems in the collection of the data—e.g., loss or gain of weight due to
preservative medium, drying times, evaporative weight loss, etc. However, biomass,
used in conjunction with species composition and abundance data, may demonstrate the
relative effects of enrichment on benthic communities (Pearson and Rosenberg, 1978).
More complicated indices — e.g., biological integrity, species diversity, species richness,
dominance, evenness — have found varying degrees of acceptance. Indices of diversity
and biological integrity often have the following limitations (Green, 1979):
• often lack biological meaning
• are not robust empirical indicators of any important correlates of "estuarine
health"
• do not incorporate information of form and function of resident species
• are susceptible to biases associated with well-described taxa
However, these indices are a widely used measure of community structure.
The dominance index is a measure of the degree to which one or a few species domi-
nate the community. The dominance index, herein defined as the minimum number of
species required to account for 75 percent of the total number of individuals, has been
useful in describing community structure (Swartz et al., 1985). It is easily calculated,
does not assume an underlying distribution of individuals among species, and is statisti-
cally testable.
-------
B-139
-\
Several graphical analyses of these simple biological indices have been developed;
most require further study before their effectiveness is known. For example, the use of
Gray and Mirza's (1979) log-normal distribution of individuals per species and
Warwick's (1985) abundance-biomass comparisons (ABC) methods, though useful for
some habitats, have been found to be unpredictable for other habitats and other locales
(U.S. EPA, 1989; Beukema, 1988).
Pearson and Rosenberg's (1978) species-abundance-biomass (SAB) model appears to
be applicable to several forms of environmental disturbance and may be used in
conjunction with other analyses to assess spatial and temporal impacts due to distur-
bance (Figure B7-1). The SAB model provides a means to assess the condition of the
habitat based on species numbers, total abundance, and total biomass (Figure B7-1).
Severely polluted areas are identified by the lack of organisms; low species numbers,
Figure B7-1.
Generalized Species-
Abundance-Biomass
diagram of changes along
a gradient of organic
enrichment (from Pearson
andRosenburg, 1978).
Increasing organic input
PO = peak of opportunists
E = ecotone point
TR = transition zone
S = species numbers
B = total biomass
A = total abundance
-------
B-140
total abundance, and biomass. Highly polluted areas are identified by a few highly-
abundant populations of small-bodied, pollution-tolerant, opportunistic species (e.g.,
Capitella capitata); total abundance is high, although biomass and species numbers
remain low. Less polluted areas (e.g., further from a point source, or over time follow-
ing the cessation of organic input) may be identified by higher species numbers and
biomass, as more competitive larger-bodied species are able to tolerate sediment
conditions. More pristine areas are characterized by a number of small populations of
competitively dominant species; species numbers remain elevated, although biomass
and total abundance are low. However, this approach requires the comparison of two
or more sites along the pollution gradient.
Indicator Species
Examination of abundances of individual indicator species are generally informative
and may reduce the cost of the analysis. The absence of pollution sensitive species
and the enhancement of opportunistic and pollution tolerant species may assist in
defining the spatial and temporal extent and magnitude of impacts. However, indica-
tor variables must possess the following characteristics (Green, 1984):
* provide sufficiently precise and accurate appraisals of:
- species of concern
- anthropogenic disturbances to benthic communities
- presence/absence or the magnitude of anthropogenic perturbance
to the estuarine ecosystem
• cost-effective and a statistically reliable alternative to monitoring all
critical benthic community measures of habitat perturbance
• appropriate for the spatial and temporal scale demanded by the study
objectives
Pearson and Rosenberg (1978) demonstrated that along a gradient of organic enrich-
ment, a predictable community of benthic infauna could be observed (Figure B7-2);
communities consist of opportunistic, tolerant species in areas of severe pollution,
giving way to less tolerant and more competitively dominant species further from
severely polluted areas (Figure B7-2). This model of indicator species composition
and distribution been found to be useful in assessing both natural and anthropogenic
disturbances.
-------
B-141
\
••••JS.-V
Several indices, derived from the distributions of pollution sensitive species and
opportunistic disturbance-tolerant species, have been developed in order to evaluate
impacts to infaunal benthic community structures: nematode/copepod ratio (Raffaelli
and Mason, 1981), infaunal index (Word, 1978), organism-sediment index (Rhoads
and Germane, 1986). The infaunal index was initially developed for the Southern
California Bight, and utilizes the abundances of four functional groupings to describe
community structure. Low values of the infaunal index indicate communities domiT
nated by deposit feeding organisms that are considered to be more impacted than
communities dominated by suspension feeders (high infaunal index values) (Word,
1978). This index has been successfully adapted for Puget Sound by Thorn et al.
(1979).
Figure B7-2. Diagram of Polluted or disturbed conditions may be indicated by:
changes in fauna and
sediment structure along a
gradient of organic
enrichment (from Pearson
and Rosenberg, 1978). • low values (< 65) of the infaunal index
high values of the nematode/copepod ratio
-------
B-142 ]
• low values (< +7) of the organism-sediment index
However, further studies of the response patterns of these indicator indices to anthro-
pogenic perturbations are required in order to select appropriate indicators of benthic
community impact.
It is recommended that multiple pollution-tolerant and pollution-sensitive indicator
species are selected by clearly defined criteria. Multiple indicator species often
provide a more complete representation of environmental conditions.
Multivariate Analyses
Numerical classification encompasses a wide variety of techniques that have been used
in the analysis of benthic infauna data to distinguish groups of entities (e.g., sample
locations) according to similarity of attributes (e.g., species). These techniques differ
from most multivariate methods in that no assumptions are made concerning the
underlying distributions of the variables. Detailed descriptions of numerical classifica-
tion analysis can be found inRomesburg (1984), Clifford and Stephenson (1975),
Bocsch (1977), Sneath and Sokal (1973), and Anderberg (1973). Boesch (1977) is
particularly valuable as an introduction and guide to the use of numerical classification
analysis in marine environmental studies. Guidance on the interpretation of classifica-
tion results is provided in U.S. EPA (1987,1988).
Ordination analyses have also been used to reduce the dimensionality (i.e., the number
of variables) of the data set while maintaining the relationship among similar and
dissimilar entities. At present no single ordination technique has been shown to be
clearly superior for the analysis of biological data (U.S. EPA, 1985).
Multivariate analyses are effective heuristic tools. They generate visual representa-
tions which often indicate where further analyses ought to be conducted.
Analytical Approach Recommendations
Some of the most informative measures of community structure are the simplest
(Table B7-2):
• number of individuals
• number of species
-------
B-143
• dominance
• infaunal index
• abundances of pollution sensitive species
• abundances of opportunistic and pollution tolerant species
These indices have proved to be useful over various habitats and regions in assessing
changes to benthic community structures (U.S. EPA, 1985). Values for these indices
may be determined from the list of species abundances generated during the taxonomic
identifications of collected specimens. Furthermore, the values of these six variables
may be easily tested statistically using parametric or nonparametric techniques. It is
recommended that no single index or analytical method be used to assess impacts;
rather the assessment of impacts should incorporate information that each variable and
method contributes concerning benthic community structure.
Selection of Reference Sites
Selection of reference sites is key to the evaluation of environmental impact assess-
ment. Monitoring of reference sites must provide measures of natural variability
against which measures of anthropogenic impacts are detected. Thus, reference sites
should be similar in every way with sampling sites of the monitoring program except
that they are unaffected by anthropogenic disturbances of concern. Results of analyses
using reference measures provide the means of discerning variability due to anthropo-
genic sources from natural variability of the ecosystem. Characteristics which should
be controlled for include:
• sediment characteristics (i.e., grain size)
• water depth
• flow characteristics
Several reference sites may be required in order to meet these criteria.
-------
B-144!
Collection B7.4
QA/QC
Sampling device should be inspected for wear and tear potentially leading to sample Considerations
leakage upon ascent. It is prudent to have a backup sampler on board the survey vessel
in case the primary sampler is found to be unsuitable during the cruise.
The following sample acceptability criteria should be satisfied (U.S. EPA, 1987a):
• sediment is not extruded from the upper face of the sampler such that
organisms may have been lost
• overlying water is present, indicating minimal leakage
• the sediment surface is relatively flat indicating minimal disturbance or
winnowing (Figure B7-3)
Acceptable if Minimum
Penetration Requirement Met
and Overlying Water is Present
Unacceptable
(Canted with Partial Sample)
Unacceptable
(Washed, Rock Caught in Jaws)
Unacceptable
(Washed)
Figure 57-3.
Examples of acceptable
and unacceptable samples
(U.S. EPA, 1986-1991).
-------
B-145
• the entire surface of the sample is included in the sampler
• the desired penetration depth is achieved
If the sample does not meet all the criteria, the sample should be rejected.
Taxonomic Identification
A key QA/QC issue is taxonomic standardization. Consistent taxonomic identifications
are achieved through interaction among taxonomists working on each major group.
Participation of the laboratory staff in regional taxonomic standardization programs is
recommended to ensure regional consistency and accuracy of identifications.
Five percent of all samples identified by one taxonomist should be re-identified by
another taxonomist who is also qualified to identify organisms in that major group. It
is advisable that at least three individuals of each taxon should be sent for verification
to recognized experts. These verified specimens should then be placed in a permanent
reference collection. All specimens in the reference collection should be stored in
labeled vials which are segregated by species and sample. Reference specimens
should be archived alphabetically within major taxonomic groups.
It is also recommended that at least 20 percent of each sample be re-sorted for QA/QC
purposes. Re-sorting is the examination of a sample or subsample that has been sorted
once and is considered free of organisms. Re-sorting should be done by someone
other than the one who sorted the original sample. If a sample is found that does not
meet the recommended 95 percent removal criterion, the entire sample should be re-
sorted.
B7.5 Consideration Of statistical strategies will mitigate the high costs of collecting and
Statistical Design processing samples. See also Statistical Design Considerations: Statistical Power and
Considerations Power-Cost Analysis (Section B.3).
Temporal Stratification of the Data
The time of the year should be controlled or stratified in the design — the use of
annual averages is seldom good practice. Temporal stratification of the data should
not be attempted until sufficient knowledge of long-term natural cycles is attained.
Initially, simple regression analyses may be conducted on seasonally stratified data in
-------
B-1461
n niiiii in HH ii mi i
"N*1 *•*••. "•, "• *v' Jf t.S1' s
order to identify monotonic temporal trends. Further examinations of whether condi-
tions are improving or degrading over time may be examined using various statistical
time series analyses.
Monitoring of benthic community structure provides in situ measures of the benthic
habitat and remains a powerful tool in the evaluation of spatial and temporal effects of
anthropogenic and natural disturbances. The presence or absence of certain infaunal
organisms are useful in indicating the previous condition of the environment. Moni-
toring of benthic infaunal communities also provides data required in the design and
validation of benthic community dynamics models (e.g., Pearson and Rosenberg,
1978) and the selection of biological indicators (e.g., Word, 1978).
In addition, monitoring of benthic communities directly provides accurate information
essential in assessing the effectiveness of pollution abatement programs (Bilyard,
1987). For example, benthic infauna monitoring provided information used to assess
the effectiveness of pollution abatement plans in the recovery of Southern California
waters (Reish, 1986; SCCWRP, 1988). These studies indicate that analyses of benthic
infaunal communities may be effectively used to monitor long-term recovery of the
receiving environment (Reish, 1986).
Currently, benthic communities have not proved useful for identifying specific chemi-
cals or classes of chemicals present in the environment. Further information concern-
ing specific responses to specific contaminants is required before infaunal community
structure becomes useful in identifying sources of contaminants (U.S. EPA, 1989). In
addition, the use of benthic community structure to predict specific effects on potential
predators is cautioned. Information on trophic relationships, competition, and preda-
tion is often not available. The capability to predict the effects of altered prey commu-
nities on predators may improve with research on these topics. Factors such as prey
quality, distribution of prey, and interactions among species will be important compo-
nents of this research.
However, benthic invertebrates serve as effective indicators of environmental condi-
tions; delineating the magnitude, spatial extent, and temporal trends of anthropogenic
and natural perturbations to the ecosystem (Reish, 1986; Bilyard, 1987). Monitoring
of benthic infauna will provide relevant and accurate data fundamental to achieving
the objectives of most estuarine monitoring programs.
B7.6
Use of Data
-------
B-147
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B7.7
Summary and
Recommendations
Rationale
• The objective is to detect and describe spatial and temporal changes in the
structure and function of benthic communities
• Provides in situ measures of habitat quality and is a powerful tool in
assessing environmental impact
Monitoring Design Considerations
• It is recommended that consistent types of sampling gear, sample sorting
. protocols, level of taxonomy, and location and timing of sample collection
be implemented to allow for comparisons among studies
• Collection of undisturbed sediment requires that the sampler:
- create a minimal pressure wave when descending
- form a leakproof seal when the sediment sample is taken
- prevent winnowing and excessive sample disturbance when
ascending
- allow easy access to the sample surface in order that undisturbed
subsamples may be taken
• Penetration well below the desired sampling depth is preferred to prevent
sample disturbance as the device closes.
• Grab samplers and box corers are recognized as the tools of choice for
maximum accuracy and precision when sampling soft-bottom habitats
• Sorting through a standard sieve mesh size is recommended - further
sorting through other mesh sizes may be conducted in addition to sorting
through this standard mesh size
• Relaxants facilitate identification and morphometric measurements,
however standard procedures must be implemented to ensure comparability
among studies
• Vital stains may facilitate sorting, however a proper QA program should
ensure that sorting efficiency is maintained
-------
B-148]
Identifications to higher taxonomic levels may be sufficient to meet
program objectives, however it is recommended that all samples be
archived since comparisons to lower taxonomic levels may be required at a
later date
• In order to reduce the variation due to seasonal differences, sampling
should be conducted during the same season - preferably the same month -
each year
Existing Analytical Methods
• It is recommended that simple measures of community structure be used to
assess the condition of the estuarine benthos: number of individuals,
number of species, dominance, infaunal index, abundance of pollution
sensitive species, and abundance of pollution tolerant species (Table B3-1)
• Selected biological indices should retain biological meaning, be robust
indicators of estuarine "health", and incorporate species form and function
• Indicator species should possess the following characteristics:
- sensitive to benthic perturbances of concern,
- cost-effective and statistically reliable alternative to measuring all
species in a monitoring program
- statistically reliable indicative measures of habitat perturbance
- appropriate for the spatial and temporal scale demanded by the
study objectives
• Selection of reference sites is key to the evaluation of environmental
impact due to anthropogenic perturbances; several reference sites may be
required to provide proper control for sampling sites
QA/QC Considerations
• Taxonomic standardization is key to the analysis of community structure -
recommended protocols include consistent interactions among taxono-
mists, re-identification of selected samples, use of a reference collection,
and re-sorting of selected samples or subsamples
-------
B-149
iV jNWAV.
Statistical Design Considerations
• Power analyses may be applied to determine the appropriate number of
sample replicates required to detect a specified difference, thereby optimiz-
ing the high costs of collecting and processing samples
Use of Data
• Provide essential information in order to assess impacts due to anthropo-
genic perturbance, monitor recovery of the receiving environment, and
validate community and population models
B7.8 Literature Amjad, S., and J.S. Gray. 1983. Use of the nematode/copepod ratio as an index of
Cited and organic pollution. Mar. Poll. Bull. 14:178-181.
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B-1501
KwwwwfWjjM. -w
L^vHP??'
I^\M^ »£«A
Bocsch, D.F. 1977. Application of numerical classification in ecological investiga-
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Elmgren, R., S. Hansson, U. Larsson, B. Sundelin, and P.D. Boehm. 1983. The
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Ferraro, S.P., F.A. Cole, W.A. DeBen, and R.C. Swartz. 1989. Power-cost efficiency
of eight macrobenthic sampling schemes in Puget Sound, Washington, USA. Can. J.
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Fleishack, P.C., AJ. DeFreitas, and R.B. Jackson. 1985. Two apparatuses for sam-
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Fredette, T.J., D.A. Nelson, T. Miller-Way, J.A. Adair, V.A. Sotler, J.E. Clausner,
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Gauch, H.G. 1982. Multivariate Analysis in Community Ecology. Cambridge, UK:
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Gray, J.S., and F.B. Mirza. 1979. A possible method for the detection of pollution
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t e Yiv «"»>. v** •<
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-¥•' 't
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,,3
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.v
f "•• "••• %,^, ^
SSrf-S^AS*. AVAV.'-'Sfc^X K V"
BS.O Fish Community Structure
B8.1 Fish are important economic, recreational, and aesthetic components of the estuarine
Rationale ecosystem. To protect and preserve healthy fish stocks, reliable estimates of fish
population abundances, a detailed knowledge of fish life histories, and information on
the effects of natural and anthropogenic disturbance are required.
The objective of monitoring fish populations is to detect and describe spatial and
temporal changes in the structure and function offish communities. These results can
be used to assess the condition - the "health" - of estuarine habitats, to monitor rates
of recovery following environmental interventions, and to provide an early warning of
potential impacts to recreational and commercial fisheries. In addition, certain demer-
sal fish are good indicators of benthic conditions since:
• they are generally sedentary - observed effects are in response to local
environmental conditions
• they are relatively sensitive to habitat disturbance - i.e., communities
undergo changes in abundance in response to environmental perturbations
• they often mediate the transfer of nutrients and toxic substances in the
ecosystem as important predator and prey organisms
Monitoring fish population abundances is one of a handful of methods which provide
in situ measures of biotic health and habitat quality. The assessment of fish popula-
tions remains a powerful tool in the evaluation of the spatial and temporal effects of
anthropogenic and natural disturbances.
B8.2
Monitoring Design
Considerations
Collection, analyses, and evaluations offish community structure and function are
typically time-consuming, labor-intensive, and expensive tasks. A survey vessel
manned by an experienced crew and specially equipped with gear to collect organisms
is required. Expert taxonomists are needed to identify and enumerate collected fish
specimens.
The results of fish monitoring programs can vary substantially depending on the
objectives and corresponding design specifications. Monitoring design characteristics
that may affect this variability include:
• type of sampling gear
• volume sampled
-------
B-1561
• location and timing of sample collection
It is essential to understand the effects of these monitoring design characteristics on the
results and to standardize them as much as possible to ensure the comparability of data
throughout the estuary.
Analyses of power-cost efficiencies are useful in selecting the appropriate sampling
gear and sample processing protocols. Ferraro et al. (1989) provide an example of
power-cost analyses.
Sampling Devices
Sample collection protocols influence all subsequent laboratory and data analysis; it is
key that fish population samples be collected using acceptable and standardized
techniques. Several types of devices can be used to collect fish samples: traps and
cages, passive nets, trawls (active nets), and photographic surveys (Fredette et al.,
1989). Many of these devices selectively sample specific types of fish. Accordingly,
conducting comparisons among data collected using different devices is inadvisable.
Traps and Cages - Traps and cages are usually designed to attract and capture
specific organisms. They are useful in studies examining the activities of a particular
target organism in a given area. Traps and cages provide only qualitative measures of
organisms in a particular area.
Passive Nets and Trawls - Nets are highly selective in the species that are captured
and in the efficiency of retaining captured specimens. The size of the net, its configu-
ration and orientation, and the avoidance behavior of the target species should be
considered when using any net. The mesh size effects the speed at which the net can
be towed. The slower the towing speed, the more likely that some organisms will
cither avoid or escape the net. It is highly recommended that the duration, direction,
and speed of towing be standardized in order to compare trawls.
Passive nets (e.g., gill nets) are deployed at a fixed position; organisms become
entangled or trapped within the netted area. Passive nets are used to collect selected
target species and provide a qualitative means of sampling fish populations. Limita-
tions associated with passive nets include:
• nets, ordinarily, must remain in-place for an extended period of time
• deployment and recovery of nets are typically time-consuming processes
-------
B-157
Trawls (active nets) are drawn through the water and results are more immediate
compared to passive nets. Trawls are typically used to collect large quantities of fish
at various depths (Fredette et al., 1989). Otter trawls are less selective than passive
nets, capturing a wider variety offish and epibenthic macroinvertebrates.
Photographic Surveys - Photographic surveys are effective when the bottom topogra-
phy is uneven or where trawling is not possible. However, the utility of photographic
surveys is limited by water clarity, difficulties identifying species, and fish avoidance
of the camera system. In addition, further studies comparing photographic surveys to
trawls are required before comparisons may be made. This method is also limited by
the qualitative nature of the data.
Volume Sampled
Different species offish have different scales of horizontal and vertical spatial distri-
bution (Gushing, 1975; Bond, 1979). Costs of laboratory analyses of the samples
increase with increased volume sampled. Analyses of spatial and temporal scale,
statistical power, as well as costs, will assist in determining optimal sample volume. It
is highly recommended that a standard sample volume (same tow duration, tow speed,
and net opening area) be analyzed in order to ensure data comparability.
Selection of Sampling Period
Fish assemblages are dynamic; the most common temporal patterns observed in fish
communities are those associated with seasonal changes. Seasonal variation in fish
assemblages may be due to changes in physical, chemical, and/or biological param-
eters: i.e., temperature, light transmissivity, dissolved oxygen, predation, recruitment,
etc.
Given the seasonal variation characteristic of fish assemblages in general, it is recom-
mended that direct comparisons,between samples collected during different seasons be
avoided. Studies investigating interannual variation in the characteristics of fish
communities should be conducted during the same season (preferably the same month)
each year.
B8.3 Analytical There are a variety of approaches to assess the effects of anthropogenic perturbance on
Methods fish communities of the estuary. These assessment approaches may be grouped into
Considerations three categories:
-------
B-1581
• biological indices
• indicator species
• multivariate analyses
However, there has been little consensus among biologists regarding the suitability of
various techniques for describing community characteristics and/or for assessing
estuarine impacts. A critical evaluation of the use of biological indices to detect
environmental change is presented in an EPA Technical Support Document (U.S.
EPA, 1985; Table B8-1). In addition to measures of change in the abundance of
pollution sensitive, pollution tolerant, and opportunistic species, the indices shown in
Table B8-1 are useful. These indices are evaluated on the basis of the following
criteria:
• biological meaning
• ease of interpretation
• sensitivity to community changes due to anthropogenic sources
The results of these evaluations and additional information on other analytical methods
are summarized below.
Biological Indices
The numbers of individuals and the numbers of species have been found to be good
indicators of anthropogenic disturbance, as well as of other environmental stresses
(U.S. EPA, 1985). Furthermore, these simple biological indices are less ambiguous
and are often as informative as diversity indices (U.S. EPA, 1985; Green, 1979;
Hurlbert, 1971). Measures of biomass have inherent problems in the collection of the
data—e.g., loss or gain of weight due to preservative medium, drying times, evapora-
tive weight loss.
More complicated indices (e.g., species diversity, species richness, dominance, even-
ness) have found varying degrees of acceptance. Diversity indices, measures of the
distribution of individuals among species, have the following limitations (Green,
1984):
• often lack biological meaning
-------
B-159
*x
TABLE B&-1. BlOLOaiCAL INDICES
Index/Method
Biological
Characteristic Measured
Recommended
for Monitoring8
Biological integrity
Bray-Curtis
Dominance15
No. individuals
No. species
Community
Dissimilarity ,
Community structure
Total abundance
.Total taxa
Opportunisticjind " Community structure
pollution tolerant species
Pollution-sensitive
species
Biomass
MargalefsSR
Pielou's 1
Shannon-Wiener H1
Community structure
Standing crop ,
Diversity
Evenness "
Diversity
B
P,B
*P (plankton), B (l)enthos), and F (fishes) indicate those biological groups to which
a givetn index may be applied, " -
^^
bDefined as the minittmtn number of spede^«e
-------
B-160 ]
• are not robust empirical indicators of any important correlates of "estuarine
health"
• do not incorporate information of form and function of resident species
• are susceptible to biases associated with well-described taxa
However, species diversity indices are a widely used measure of community structure.
The dominance index is a measure of the degree to which one or a few species domi-
nate the community. The dominance index, herein defined as the minimum number of
species required to account for 75 percent of the total number of individuals, has been
useful in describing community structure (Swartz et al., 1985). It is easily calculated,
does not assume an underlying distribution of individuals among species, and is
statistically testable.
Indicator Species
Examination of abundances of individual indicator species are generally informative
and may reduce the cost of the analysis. The absence of pollution sensitive species
and the enhancement of opportunistic and pollution tolerant species may assist in
defining the spatial and temporal extent and magnitude of impacts. However, indica-
tor variables must possess the following characteristics (Green, 1984):
• provide a sufficiently precise and accurate appraisals of:
- specie(s) of concern
- anthropogenic disturbances to benthic communities
- presence/absence or the magnitude of anthropogenic perturbance
to the estuarine ecosystem
• cost-effective and a statistically reliable alternative to monitoring all
critical community measures of habitat perturbance
• appropriate for the spatial and temporal scale demanded by the study
objectives
Further studies of the response patterns of fish species subjected to anthropogenic
perturbations are required in order to select appropriate indicators of environmental
impact. When a suitable indicator species is identified, it is desirable to monitor the
-------
B-161
status and trend of that species' population. This will also be true of certain species
having high economic or public value (e.g., striped bass). A wide variety of methods
can be used to measure the size offish populations and assess population structure.
These include mark and recapture techniques and various techniques used to determine
population sex and age structure. Nielsen and Johnson (1984) and Cailliet et al.
(1986) provide thorough discussions of these techniques and their applications. Ricker
(1975) provides an excellent discussion of methods for the sampling of fish popula-
tions.
Multivariate Analyses
Numerical classification encompasses a wide variety of techniques that have been used
in the analysis offish data to distinguish groups of entities (e.g., sample locations)
according to similarity of attributes (e.g., species). These techniques differ from most
multivariate methods in that no assumptions are made concerning the underlying
distributions of the variables. Detailed descriptions of numerical classification analy-
sis can be found in Romesburg (1984), Clifford and Stephenson (1975), Boesch
(1977), Sneath and Sokal (1973), and Anderberg (1973). Boesch (1977) is particularly
valuable as an introduction and guide to the use of numerical classification analysis in
marine environmental studies. Guidance on the interpretation of classification results
is provided in an EPA Technical Support Document (U.S. EPA, 1988).
Ordination analyses have also been used to reduce the dimensionality of the data set
while maintaining the relationship among similar and dissimilar entities. At present no
single ordination technique has been shown to be clearly superior for the analysis of
biological data (U.S. EPA, 1985).
Multivariate analyses are effective heuristic tools. They generate visual representa-
tions which often indicate where further analyses ought to be conducted.
Analytical Approach Recommendations
Some of the most informative measures of community structure are the simplest
(Table B8-1):
• number of individuals
• number of species
• dominance
-------
B-1621
•,; j
• abundances of pollution sensitive species
• abundances of opportunistic and pollution tolerant species
These indices have proved to be useful over various habitats and regions in assessing
changes to fish community structures (U.S. EPA, 1985). Values for these indices may
be determined from the list of species abundances generated during the taxonomic
identifications of collected specimens. Furthermore, the values of these six variables
may be easily tested statistically using parametric or nonparametric techniques. It is
recommended that no single index or analytical method be used to assess impacts;
rather the assessment of impacts should incorporate information that each variable and
method contributes concerning benthic community structure.
Selection of reference sites is key to the evaluation of environmental impact assess-
ment.
Results of analyses using reference measures provide the means of comparison by
which anthropogenic impacts are detected. It is essential that selected reference sites
exhibit similar:
• bottom characteristics (i.e., bottom topography, sediment grain size,
percent cover)
• water depths
• flow characteristics
compared to monitoring program sampling sites. Several reference sites may be
required in order to meet these criteria.
Sample Collection
The primary consideration for an adequate sampler is that it consistently collect
undisturbed samples. Nets should be inspected for wear and tear potentially leading to
sample loss. It is prudent to have backup gear on board the survey vessel in case the
primary gear is found to be unsuitable or lost during the cruise.
B8.4
QA/QC
Considerations
-------
B-163
Taxonomic Identification
A key QA/QC issue is taxonomic standardization. Consistent taxonomic identifica-
tions are achieved through interaction among taxonomists working on each major
group. Participation of the laboratory staff in regional taxonomic standardization
programs is recommended to ensure regional consistency and accuracy of identifica-
tions.
The verified specimens should then be placed in a permanent reference collection. All
specimens in the reference collection should be stored in labeled vials which are
segregated by species and sample. Reference specimens should be archived alphabeti-
cally within major taxonomic groups.
B8.5 Consideration of statistical strategies will mitigate the high costs of collecting and
Statistical Design processing samples. See also Statistical Design Considerations: Statistical Power and
Considerations Power-Cost Analysis (Appendix B Introduction; Section B.3).
Temporal Stratification of the Data
The time of the year should be controlled or stratified in the design; the use of annual
averages is seldom good practice, especially for species which are seasonally abun-
dant. Temporal stratification of the data should not be attempted until sufficient
knowledge of long-term natural cycles is attained. Initially, simple regression analyses
may be conducted on seasonally stratified data in order to identify monotonic temporal
trends. Further examinations of whether conditions are improving or degrading over
time may be examined using various statistical time series analyses (e.g., temporal
autocorrelation, spectral analyses).
B8.6
Use of Data
Monitoring of fish community structure provides in situ measures of the estuarine
habitat and remains a powerful tool in the evaluation of spatial and temporal effects of
anthropogenic and natural disturbances. The presence or absence of certain fish
species is useful in indicating the condition of the environment. Monitoring of fish
communities also provides data required in the design and validation of fish population
dynamics models and the selection of biological indicators (U.S. EPA, 1990). Fishing
mortality, both commercial and sport, is an important source of anthropogenic impact
on fish populations. Before the effects of pollution can be assessed, fishing impacts
must be considered. There are several methods available to assess the effects of
fishing mortality, allowing pollution impacts to be determined (Ricker, 1975).
-------
B-1641
In addition, monitoring offish communities may directly provide accurate information
essential in assessing the effectiveness of pollution abatement programs. Analyses of
fish communities may be effectively used to monitor long-term recovery of the receiv-
ing environment (Gushing, 1975).
Fish serve as effective indicators of environmental conditions; delineating the magni-
tude, spatial extent, and temporal trends of anthropogenic and natural perturbations to
the ecosystem. Monitoring of fish populations will provide relevant data fundamental
to achieving the objectives of many estuarine monitoring programs.
Rationale
• The objective is to detect and describe spatial and temporal changes in the
structure and function offish communities in order to protect the eco-
nomic, recreational, and aesthetic value of the estuarine fisheries
• Provides in situ measures of habitat quality and is a powerful tool in
assessing environmental impact
Monitoring Design Considerations
• It is recommended that consistent types of sampling gear, volume sampled,
and location and timing of sample collection be implemented to allow for
comparisons among studies
• In order to reduce the variation due to seasonal differences, sampling
should be conducted during the same season - preferably the same month -
each year
Existing Analytical Methods
• It is recommended that simple measures of community structure be used to
assess the condition of the estuarine fish: number of individuals, number of
species, dominance, abundance of pollution sensitive species, and abun-
dance of pollution tolerant species (Table B8-1)
• Selected biological indices should retain biological meaning, be robust
indicators of estuarine "health", and incorporate species form and function
B8.7
Summary and
Recommendations
-------
B-165
-ssr-
• Indicator species should possess the following characteristics:
- sensitive to perturbances of concern
- cost-effective and statistically reliable alternative to measuring all
species in a monitoring program
- statistically reliable indicative measures of habitat perturbance
- appropriate for the spatial and temporal scale demanded by the
study objectives
• Selection of reference sites is key to the evaluation of environmental
impact due to anthropogenic perturbances; several reference sites may be
required to properly control for sampling sites
QAIQC Considerations
• Taxonomic standardization is key to the analysis of community structure -
recommended protocols include consistent interactions among
taxonomists, re-identification of selected samples, and use of a reference
collection
Statistical Design Considerations
• Power analyses may be applied to determine the appropriate number of
sample replicates required to detect a specified difference, thereby optimiz-
ing the high costs of collecting and processing samples
Use of Data
• Provide essential information in order to assess impacts due to anthropo-
genic perturbance, monitor recovery of the receiving environment, and
validate community and population models
B8.8
Literature Cited
and References
Anderberg, M.R. 1973. Cluster Analysis for Applications. New York, NY: Aca-
demic Press. 359pp.
Boesch, D.F. 1977. Application of numerical classification in ecological investiga-
tions of water pollution. EPA 600/3-77-033. Office of Research and Development,
US Environmental Protection Agency. Corvallis, OR. 115pp.
-------
B-1661
\
'/J
Bond.CJS. 1979. Biology of Fishes. Philadelphia, PA: Sanders College Publishing.
514pp.
CaSffiet, G.M., M.S. Love and A.W. Ebeling. 1986. Fishes: A field and laboratory
manual on their structure, identification, and natural history. Belmont, CA:
Wadsworth Publishing Company.
Clifford, H.T. and W. Stephenson. 1975. An introduction to numerical classification.
New York, NY: Academic Press. 229pp.
Gushing, D.J. 1975. Marine Ecology and Fisheries. Cambridge, U.K.: Cambridge
University Press. 278pp.
Fcrraro, S.P., F.A. Cole, W.A. DeBen, and R.C. Swartz. 1989. Power-cost efficiency
of eight macrobenthic sampling schemes in Puget Sound, Washington, USA. Can. J.
Fish. Aquat. Sci. 46:2157-2165.
Fredette, T.J., D.A. Nelson, T. Miller-Way, J.A. Adair, V.A. Sotler, J.E. Clausner,
E.B. Hands, and F. J. Anders. 1989. Selected tools and techniques for physical and
biological monitoring of aquatic dredged material disposal sites. Final Report. U.S.
Army Engineer Waterways Experiment Station, Vicksburg, MS.
Gauch, H.G. 1982. Multivariate Analysis in Community Ecology. Cambridge, UK:
Cambridge University Press. 298 pp.
Green, R.H. 1979. Sampling Design and Statistical Methods for Environmental
Biologists. John Wiley & Sons, NY. 257pp.
Green, R.H. 1984. Statistical and nonstatistical considerations for environmental
monitoring studies. Environ. Monitoring and Assessment 4:293-301.
Hurlbert, S.H. 1971. The nonconcept of species diversity: A critique and alternative
parameters. Ecol. 52:577-586.
Hurlbert, S.H. 1984. Pseudoreplication and the design of ecological field experi-
ments. Ecol. Monogr. 54:187-211.
Nielsen, L.A. and D.L. Johnson (eds). 1984. Fisheries techniques. 496pp. Bethesda,
MD. American Fisheries Society.
-------
B-167
Ricker, W.E. 1975. Computation and interpretation of biological statistics offish
populations. Bull. Fish. Res. Bd. Can. 191:382 pp.
Romesburg, H.C. 1984. Cluster Analysis for Researchers. Belmont, CA: Lifetime
Learning Publications. 334pp.
Self, S.G., and R.H. Mauritsen. 1988. Power/sample size calculations for generalized
linear models. Biometrics. 44:79-86.
Sneath, P.H.A. and R.R. Sokal. 1973. Numerical Taxonomy: The Principles and
Practices of Numerical Classification. San Francisco, CA: Freeman. 573pp.
Swartz, R.C., D.W. Schultz, G.R. Ditsworth, W.A. DeBen, and F.A. Cole. 1985.
Sediment toxicity, contamination, and macrobenthic communities near a large sewage
outfall. Tiv Validation and Predictability of Laboratory Methods for Assessing the Fate
and Effects of Contaminants in Aquatic Ecosystems. (T.T. Boyle, ed.). Philadelphia,
PA: American Society for Testing and Materials (ASTM). pp. 152-175.
U.S. EPA. 1978. Use of small otter trawls in coastal biological surveys. EPA
600/3-78-083. Office of Research and Development, Corvallis, OR. 35pp.
U.S. EPA. 1985. Recommended biological indices for 301 (h) monitoring programs.
EPA 430/9-86-002. Of fice of Marine and Estuarine Protection, Washington, DC 34pp.
U.S. EPA. 1987a. Technical support document for ODES statistical power analysis.
EPA 430/9-87-005. Office of Marine and Estuarine Protection, Washington, DC. 34 pp.
U.S. EPA. 1987b. Puget Sound protocols. Final Report. 31pp. Prepared for Region X,
Office of Puget Sound.
U.S. EPA. 1988. ODES data brief: Use of numerical classification. Prepared for the
Office of Marine and Estuarine Protection. 18pp.
U.S. EPA. 1990. Environmental Monitoring and Assessment Program: Ecological
Indicators. EPA 600/3-90-060. Office of Research and Development, Washington, DC.
-------
B-1681
s\\%>\sy,
-------
B-169
B9.0 Fish and Shellfish Pathoblology
Pathobiological methods provide information concerning damage to organ systems of
fish and shellfish through an evaluation of their structure, activity, and function.
Anatomic pathology methods can give an indication of the nature of an altered state,
for example, by identifying the specific type of tumor present in an animal. Repro-
ductive developmental studies examine the reproductive capacity of animals and can
provide information to aid in estimating and predicting population abundance and
recruitment. Biochemical/enzymological studies seek to detect differences in enzy-
matic activity as a measure of biological condition. Immunological methods can
demonstrate altered immune response, an indicator of changes in bodily defense
mechanisms and susceptibility to disease.
Pathobiological methods should be used in concert to investigate cause and effect
relationships as a result of contaminant exposure. Anatomic pathology can serve as a
vital link between observed effects on populations and communities in an estuary and
the changes in activity and function observed by other methods.
B9.1 Pathobiological methods can be used to examine adverse effects of pollutants on fish
Rationale and shellfish. The presence of toxics in water and sediments may not immediately
result in visible changes in these organisms. Biomarkers offer a more sensitive and
reliable assessment of exposure risks than ambient water or sediment quality monitor-
ing. Monitoring of pathobiological effects provide information necessary to make
determinations of the existence of adverse effects in animals (e.g., tumors), population
productivity and stability (affected by reproduction and disease states) and the loss of
organisms deemed valuable for ecological, aesthetic, recreational, scientific or eco-
nomic reasons. Table B9-1 outlines some of the terms used to describe
pathobiological methods.
Although the value of these methods for establishing cause and effect links has been
established during laboratory toxicity studies, some questions remain regarding their
ability to establish such links for field-collected organisms that are exposed to a variety
of natural environmental stresses and combinations of contaminants (Hinton and
Couch, 1984; Couch and Harshbarger, 1985; Mix, 1986; Sindermann, 1991). How-
ever, properly conducted multidisciplinary monitoring studies using these methods can
provide regulatory agencies with evidence of impaired health status in animals ex-
posed to contaminants in estuarine ecosystems. This information can then be used to
direct laboratory confirmation of the cause, if necessary (see for example, Buckley
et al., 1985; Gardner et al., 1991). Continued monitoring with these methods can be
used to detect changes in a population's health during and following environmental
interventions. Because changes at the organismal level precede changes in population
-------
B-170!
.
j V1, % , "•> . "•*• *• fSS ,; S * ?"" SS\f
^W^^^^^
TABLE B9-1. LIST OF PATHOBIOLOGICAL TERMS
biomarkcr
cytochromeP450
cytogenctics
genotoxic agents
hepatic
histopathology
histocherriistry
immunoassay
immunology
inclusion bodies
macrophage
pathobiology
•rt i -•^ -;s
- any biologjcal tneflioduised to detect the exposure of prganism$ to hawdous chemicals m flio
environment by measuring tEe response of the organisms to ttie; - s "
m a protein'in themiCroSOmeS of liver cells which is important in catalyzmg the metabolism of s
steroid hc»Tnones"and fatty acids an8 in thedetoxijScafion of a vantety of chemical substances
= the study of cytology in telatibn%to genetics> especially the study of ch'irc^osomal behavior in
mitosis and meiosis. Modem cy togenetics has led to the identification of chromosomes as
bearers of the genes and^deoxyribonucleic acid (DNA) as the key molecule of the gene
= chemicar'a/id physical agehtsjhat can jprodwe genetic alterations at subtoxic concentrations
and which can f&Sult in altered heredity characteristics. G^notoxic agents generally possess
specified cbemfelfl or physical p'roperties that facilitate their interaction with nucleic ackte
= of, or relatrn|tor&e livery , "'t ^ " ""\^^" •
^ Vk»'« =» "• '•''•^•«, V *" ^ X% .,.•-• -w V V V, N
= pathologic hislology; the sciehc^ ^study dealing with the cytologto and h jstologic (mjcrch
sCOpic) SttuCtur&" Of abnormal Or Diseases cells, tissues^, and Organs in rlla&On to thefr function"
%v*\; % : > ,, / f ^ ^ t *^- ^;;\
« the study of tee chemfetry of cells and ^ssues using light and electron,miCrOscopy, special
chemica^teststani special stams to,determine the location of certain enzyme systems c*
reaction productl;in the cell " "^"x% v "-^^ * '"
-^ •" ffff*"' * * * '"*' f " *••<•*... •• '
~° ~-\ ^
=* measuring the protein ajii protein-bburid molecule^ that are cbncemed with the reaction of an
antigen withlfs speciticlintibody, as in the detecSol^of hormones or other substances
= the snidy of being protected from a diseases the study of the response of the body andJtr -
tissues to £ variety of antigensr including red Cells, pollens^ transplanted tissues and even the
individual's own cells " ; % ''"-.- -" "
- bodies present in'ttie nuclear or cytoplasm of certain cells in 'cases of infection by filtrable
viruses or ds'the result of degenerative diseases Or exposure to chemicals
» cells of thfe reticuloendothelial system having the ability to phagoCylOSe paniculate substances
and to storexvitalsdyesand^other%coUoidalsubstances
smooth cndoplasrnic
reticulum (SER)
= pathology, the study of disease} with emphasis more on the biological than on the medical
aspects of theessential natuteTdauses; and development of abnormal conditions, as well as di
structural and^f unctlonal changes that result from the disease process
/\ V;^- r v , ,„ :%:„ , •••"" , "~ .
a connectingnetwor^oFtuijuIes that course through tiiecytoplasm oftheceU and which, can
be viewed using l%electrpn microscope and is essential to metabolic JFuncfionsOf the cells
(Stedman's Medical Dictionary, 1982; Taber's Cyclopedic Medical Dictionary, 1985)
-------
B-171
and community characteristics, pathobiological studies can provide an early indication
of the effectiveness of management actions.
B9.2
Monitoring Design
Considerations
A field survey to collect target organisms and tissue samples may be required for
pathobiological monitoring (Hargis et al., 1984; U.S. EPA, 1987a). In certain in-
stances, a large sample size may be needed to establish statistical significance because
of normal variation from animal to animal, species and generic differences, and
migratory habits of fish. For these reasons, and the labor-intensive nature of
pathobiological methods, the availability and allocation of funds, time, equipment, and
trained personnel must be considered when planning to include pathobiological
methods in monitoring programs.
Selection of Target (Indicator) Species
A key component of any pathobiological monitoring program is the selection of target
species. The fundamental criterion is the ability to use the selected species to make
comparisons between sampling locations and sampling periods.
It is recommended that the target species possess the following characteristics:
• abundant enough, temporally and spatially, to allow for adequate sampling
• large enough to provide adequate amounts of tissue for analysis
• sedentary (non-migratory) in nature to assure that pathobiological abnor-
malities are representative of the study area
• easily collected
• existing database of exposures and sensitivities
U.S. EPA has provided a list of suggested fish target species for different regions in
the United States (U.S. EPA, 1987a; Table B9-2). Fish provide sufficient tissue for
analyses and may indicate potential threats to human populations. However, fish are
motile and pathobiological abnormalities detected may not be representative of the
study area.
Bivalve molluscs, either attached to the substratum or burrowing in sediments, have
been sampled extensively to examine the condition of these commercially important
-------
B-172
s *•««.
TABLE B9-2. HIGHEST RANKING CANDIDATE FISHES FOR USE
AS PATHOBIOLOGY MONITORING SPECIES
Secondary Selection Criteria
State
Locality
Species
Economic
Importance
MASSACHUSETTS Swampscott
Lynn
South Essex
Boston
Fall River
New Bedford
RHODE ISLAND Newport
NEW YORK Upper East River
Lower East River
Lower Hudson River
NEW JERSEY Cape May
VIRGINIA
Portsmouth
Virginia Beach
Winter flounder
Yellowtail flounder
Ocean pout
Windowpane
Winter flounder
Yellowtail flounder
Ocean pout
Winter flounder
Yellowtail flounder
Windowpane
American eel
Ocean pout
Winter flounder
Yellowtail flounder
Ocean pout
Windowpane
Winter flounder
Windowpane
Winter flounder
Scup
Summer flounder
Winter flounder
Scup
Weakfish
Winter flounder
Windowpane
Weakfish
Spot
Scup
American eel
Hogchoker
Spot
Red hake
Windowpane
Summer flounder
Spot
Summer flounder
Atlantic croaker
Hogchoker
Spot
Red hake
Summer flounder
Yes
Yes
No
No
Yes
Yes
No
Yes
Yes
No
No
No
Yes
Yes
No
No
Yes
No
Yes
Yes
Yes
Yes
Yes
Yes
Yes
No
Yes
Yes
Yes
No
No
Yes
No
No
Yes
Yes
Yes
Yes
No
Yes
No
Yes
Bioassay
Species
Yes
Yes
No
No
Yes
No
No
Yes
No
No
No
No
Yes
No
No
No
Yes
No
Yes
No
Yes
Yes
No
No
Yes
No
No
Yes
No
No
No
Yes
No
No
Yes
Yes
Yes
No
No
Yes
No
Yes
-------
B-173
••Su
V
TABLE B9-2.
(continued)
Secondary Selection Criteria
State
CALIFORNIA
(NORTHERN)
CALIFORNIA
(SOUTHERN)
WASHINGTON
Locality
San Francisco
Oakland
Monterey
Santa Cruz
Watsonville
Goleta
Santa Barbara
L.A. County
Orange County
Hyperion
Oceanside
Escondido
San Elijo
San Diego
Central Puget Sound
Species
English sole
Pacific sanddab
Big skate
English sole
Starry flounder
Pacific staghorn sculpin
English sole
Curlfin sole
English sole
English sole
Curlfin sole
Dover sole
Pacific sanddab
Longspine combfish
Spotted cusk-eel
English sole
Pacific sanddab
Dover sole
Curlfin sole
English sole
Dover sole
Pacific sanddab
English sole
Dover sole
Longspine combfish
Big skate
California skate
Dover sole
Blackbelly eelpout
Pacific sanddab
English sole
Dover sole
English sole
Pacific sanddab
Longspine combfish
English sole
Dover sole
Rock sole
Spotted ratfish
Rex sole
C-O sole
Economic
Importance
Yes
Yes
No
Yes
Yes
No
Yes
Yes
Yes
Yes
Yes
Yes
Yes
No
No
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
No
No
No
Yes
No '
Yes
Yes
Yes
Yes
Yes
No
Yes
Yes
Yes
No
Yes
Yes
Bioassay
Species
Yes
No
No
Yes
No
No
Yes
No
Yes
Yes
No
No
No
No
No
Yes
No
No
No
Yes
Yes
Yes
Yes
No
No
No
No
No
No
No
Yes
No
Yes
No
No
Yes
No
No
No
No
No
-------
B-174
cstuarine species (i.e., Mussel Watch, National Status and Trends program). The most
common target species have been oysters (Crassostrea virginica) and mussels (Mytilus
spp.)i although pathobiological and bioassay studies have also been performed on
other species, such as crabs and penaeid shrimp (see also Table B10-3).
Secondary considerations, based on economic importance and status as a bioassay
organism, may be applied to further winnow the list of candidate target species to a
practical number of species to be analyzed.
Sampling Location
Appropriate locations of sampling stations depend on the objectives of the study. For
example, to evaluate whether there is a statistically significant increase in lesions,
stations should be located to collect specimens from contaminated and uncontaminated
(background or control) areas for statistical comparison. It is also important to demon-
strate a dose-response relationship between the pollutant concentration and tumor
incidence by locating sample stations along a contamination gradient (i.e., from highly
contaminated to moderately contaminated to uncontaminated). It is recommended that
statipns be located in areas where the geographic area of contamination is large enough
that sampled fish could reasonably be expected to have spent a considerable amount of
time within the influence of the pollutant (U.S. EPA, 1987a). Nielsen and Johnson
(1984) and Cailliet et al. (1986) may be consulted for further fish sampling method-
ologies. Information on sampling bivalves and other invertebrates is contained in
Couch (1978), Yevich and Barszcz (1983), Turgeon et al. (1991), and other sources.
Anatomic Pathology Methods
Anatomic pathology methods examine tissues with the naked eye (gross anatomic
pathology), the aid of a light microscope (LM methods), or the electron microscope
(EM methods). Gross anatomic methods are concerned with obvious adverse changes
in tissue which can be observed in the field (Hunn, 1988; Hargis et al., 1984). The
advantage of gross methods is that large numbers of specimens can be examined
rapidly. However, the methods are generally nonspecific (i.e., it is not possible to
determine that a specific pollutant led to a specific disease). An exception to this is
cataracts in fish, which have been linked with polynuclear aromatic hydrocarbon
pollution in the field (Hargis and Zwemer, 1988).
A disadvantage of anatomic pathology methods is that they require specialized person-
nel and laboratories. The methods are, however, generally standardized, routine, and
B9.3
Existing Analytical
Methods
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B-175
operational in existing federal, state, university, veterinary, and private diagnostic
laboratories that specialize in aquatic animal pathology. Current activities are aimed at
developing a field-to-laboratory response-diagnostic scenario where adverse effects
are found in the field, diagnosis is made in the laboratory, and experimental studies are
initiated to verify results (May, 1990). See Yevich and Barszcz (1980), Howard and
Smith (1983), Johnson and Bergman (1984), Klontz (1985), Meyers and Hendricks
(1985), and U.S. EPA (1987a) for more information on these methods and techniques.
Light Microscopy Methods - LM (histologic) methods use the light microscope to
view cells and tissues. These methods can detect changes such as inflammatory
responses, alterations in the appearance of cells, cancerous/precancerous lesions, and
damage due to parasites (U.S. EPA, 1986). The advantages of LM methods are that
they are organ specific (i.e., the lesion can be localized to a specific organ) and can
detect microscopic and subtle cellular alterations that are not evident on visual inspec-
tion. However, LM methods are more expensive (approximately $30/sample) and
slower than gross methods (May, 1990).
Electron Microscopy Methods - EM methods use the electron microscope to detect
changes in tissue at cellular and subcellular levels, such as the identification of the
nature of inclusion bodies or changes in the amount of smooth endoplasmic reticulum.
An advantage of EM methods is that they can be highly specific because the pollutant
can be localized within certain parts of the cell. These inclusions may contain the
causative chemical agent; lead, gold, iron, bismuth, uranium, beryllium, mercury,
copper and arsenic are a few of the metals that can be deposited intracellularly
(Sorenson and Smith, 1981). Additionally, EM methods can be used to investigate
subcellular mechanisms of pollutant action. The major disadvantage of EM methods is
that they are very expensive (minimum $400/fish) and very slow, requiring highly
skilled technical expertise.
Histochemical Methods - Histochemical methods use the microscope in conjunction
with special chemical tests and stains to localize specific enzyme systems or reaction
products in the cell. For instance, histochemical assays have been used on liver tissue
during and after tumor formation to yield important information on the biochemistry of
specific lesions (Hinton etal., 1988; Luna, 1968; Sumner, 1988). Histochemical
methods are reliable and can be highly specific for certain classes of organic com-
pounds and metals. However, highly skilled technical expertise is required to carry out
the methods in the laboratory.
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B-176 J
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In vitro Tests -//i v/fro tests are generally more sensitive than whole animal systems,
less expensive to carry out and of shorter duration. However, in in vitro tests, defense
mechanisms found in the intact animal are missing. In vitro systems offer the greatest
flexibility for the testing and study of environmental contaminants. They can be
designed to be relevant to the species of interest in a given area, and multiple types of
measurements can be taken from a single test system (e.g., metabolic products, mitotic
activity, cytotoxicity and genetic damage). However, comparative in vitro and in vivo
studies are needed to correlate and relate changes that occur in each system (Landolt
and Kocan, 1983).
Reproductive/Developmental Methods
Reproduction studies are designed to examine a number of parameters in the reproduc-
tive process: gross examination of the egg and numbers of eggs, embryo viability, the
proportion of fertilized eggs (i.e., fertilization success), and larval development and
viability (U.S. EPA, 1986). Studies that examine the egg itself incorporate micro-
scopic methods which are time-consuming and expensive (West, 1990), but serve as a
direct measure of reproductive success. Some methods deal with the egg at the
molecular level and analyze the action of chemical and physical agents whose toxicity
is directed toward genetic (DNA) components of the egg.
Different cytogenotoxic tests can be used to measure a diverse array of effects includ-
ing gene mutation, chromosome damage (sister chromatid exchange), primary DNA
damage, or oncogenesis (i.e., tumor formation and development) (Brusick, 1980;
Landolt and Kocan, 1983; Klingerman, 1982; Shugart, 1990). Another area that is
currently being investigated as a suitable, sublethal assay for toxicity of certain wastes
in the marine environment is the fin regeneration test (Weis et al., 1990). The infor-
mation collected using these methods can be used to assess the effects of pollutants on
the reproductive capacity of animals. An understanding of these effects can be useful
in evaluating observed population and community level changes relative to the occur-
rence of specific pollutants.
Gonadotropic and steroidogenic hormones regulate the reproductive capacity of an
organism. The level of these hormones have been used to assess how pollutants affect
the reproductive capacity offish (Veranasi, 1990). The analysis is a sensitive indicator
of exposure affecting major biological processes that impact the whole population.
However, relatively detailed information about the normal reproductive cycle of the
animals is necessary to apply these methods in the field (Veranasi, 1990).
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Biochemical Methods
Biochemical methods have been used in field studies to measure various indicators of
environmental contamination. These methods are inherently sensitive and may
provide basic information about early changes in response to environmental contami-
nation at the cellular level. The development of a suite of indicators having both
specific and nonspecific responses can provide information on the type of stressors,
mechanisms of action, the extent of physiological dysfunction, and potential long-term
population consequences (Thomas, 1990).
Fish can respond to generalized stress, contaminants being one type of stress, through
induction (increased synthesis) of stress proteins (Pickering, 1981; Sanders, 1990).
Stress proteins are currently being investigated for use as generalized biochemical
indicators of stress in fish, chemical-class pollutant indicators, and mode of action
indicators. The methods for detecting stress proteins involve radioisotopic and immu-
nologic methods that measure the amount of stress protein present after a stress (i.e.,
exposure to a pollutant) occurs. At present, cDNA probes are being used experimen-
tally to measure the correlation between stress and induction of stress proteins. These
methods potentially afford a high degree of sensitivity.
It has been suggested that induction of the fish hepatic microsomal mono-oxygenase
(MO) enzyme could serve as a sensitive biological indicator for certain classes of
chemicals in water (Payne et al., 1987; Kleinow et al., 1987; Lech et al., 1982; ,
Jimenez et al., 1990; Haux and Fo'rlin, 1988). Metallothioneins (MT) have been under
consideration for use as a monitoring tool for trace environmental metal pollution due
to their induction as a result of exposure to certain metals (Engel and Roesijadi, 1987;
Garvey, 1990; Haux and FOrlin, 1988). However, additional scientific research is
required to understand the basic biology of fish before the exact significance of field
studies using these techniques can be ascertained.
A concern when measuring biochemical variables in fish to detect environmental
pollutants is that their exact biological significance is rarely understood. In addition,
for most of the biochemical variables studied, the normal range for a particular fish
population and the factors influencing these variables are often unknown (Neff, 1985).
Even with these limitations, biochemical methods hold considerable promise as
sensitive early indices of exposure to environmental stressors (Thomas, 1990). How-
ever, additional research is needed so that simplified, more cost effective field method-
ologies can be developed.
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B-178!
Immunological Methods
Immunological biomaricers are simple, sensitive, reproducible, and workable in the
field (Weeks et a/., 1990; D. Anderson, 1990a; R.S. Anderson, 1990). These indica-
tors provide supportive evidence for linkage between a stressor (toxicant, etc.) and
disease outbreaks in fish and shellfish. The immune response can be used to monitor a
specific antigen or microorganism responsible for pathological conditions in fish.
Biologists can perform quick and sensitive assays in the field or in their own diagnos-
tic laboratories because many immune assays are becoming available in kits (Rowley,
1990; Matthews et al., 1990). Many immunological assays do not require sacrifice of
the animal. Blood samples can be taken periodically to follow the kinetics of the
effects of stress in a single animal; however, the effects of handling stress on aquatic
species must also be considered in this case. The immune response is physiologically
similar among most vertebrates and similar equipment and materials can be used to
test all species offish as well as shellfish (see Anderson, 1987). There is a rapidly
growing literature on immunotoxicology from veterinary and aquatic animal sciences.
The selection of immune system indicators for the study of stress effects depends on
many factors, including specific study objectives, available equipment, training,
personnel, and length and number of assays. The most sophisticated and sensitive
assays are costly and require highly trained personnel, whereas simple assays can be
performed by field biologists with only basic laboratory supplies.
A major limitation of immune indicators is that the response is sometimes too broad to
provide conclusive evidence that the observed reaction is actually due to a specific
complex to be considered. Cross-reactions and heightened responses to nonspecific
factors may prevent the interpretation of assays with absolute certainty. The immune
response in fish or shellfish will be distinctive for a specific antigen or disease-causing
agent This will frequently make it difficult to know which immune indicator is most
affected and which immunological assay to apply. Expensive materials and laboratory
equipment are needed for sophisticated immunological assays. Confirmatory assays
are advisable to be sure that a particular stressor is the only cause of a particular effect.
Many animals should be sampled because of natural variability among individuals.
Physiological Methods
Hematological methods have been used by biologists for many years to assess the
general health offish in hatcheries and research laboratories. The procedures are well-
standardized (Blaxhall and Daisley, 1973; Wedemeyer and Yasutake, 1977) and are
easy to carry out, even in the field. Hematological measures may be affected by the
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S f
stress of capture, but are far less influenced than some other measurements, such as
blood glucose (Larson et al, 1985). Hematologic methods have been successfully
implemented in the field and have been rated as the best physiologic method for
evaluation of pollutants (U.S. EPA, 1986). For the measurement of hematological
factors in fish, techniques similar to those used in human and veterinary clinical
laboratories are generally used with minor modifications (Heath, 1987; Bouck 1984).
Blaxhall and Daisley (1973), Wedemeyer and Yasutake (1977), and Ellis (1977)
provide practical guides to the adaptation of these methods for use on fish blood.
The hematocrit/erythrocyte determination may not be as sensitive to pollution as is the
leucocyte count, at least as far as its response to metals is concerned (Larson et al.,
1985). It remains to be determined how sensitive the procedure is to subtle environ-
mental changes. In general, chemical and physical stressors cause a decrease in the
leucocrit, whereas infections produce the opposite response. However, it is also
possible to get elevations in granulocytes concomitant with a decrease in lymphocytes,
thereby yielding an unchanged leucocrit (Peters et al., 1980). Thus, the method has its
limitations for the detection of chronic stress (Wedemeyer et al., 1983).
There are several obstacles, such as capture stress, that limit the potential usefulness of
physiological tests in field work. Capture stress precludes the use of sensitive, early
indicators of environmental stress. The use of physiological responses of wild fish to
assess environmental quality is difficult because responses due to toxicants often
cannot be distinguished from those induced during handling of the wild fish (Bouck,
1984).
B9.4
QA/QC
Considerations
Pathobiological methods have a wide range of sensitivities and response times (i.e.,
when a response can be detected). For instance, certain immunologic, electron micro-
scopic, and biochemical methods can detect early changes in cells and are very sensi-
tive. Gross anatomic pathology methods, on the other hand, can only detect cellular
changes after lesions can be seen and, therefore, are less sensitive.
General considerations for QA/QC procedures have been covered earlier in this
document (Section B.I). With regard to pathobiological methods, it should be noted
that careful and consistent handling of aquatic specimens is required to minimize
trauma and confounding effects, such as exposure to air. Organisms should be held in
the laboratory under conditions as near to those found at the site of collection as
possible and for as short a time as practicable before performing assays. Fish and
shellfish collected for histopathological examination must be properly fixed (e.g.,
immersed in a formaldehyde or glutaraldehyde solution) to stop metabolic activity.
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B-1801
This may require opening or sectioning the organisms to allow the fixative to rapidly
penetrate all tissues and preserve the cellular structure in its existing condition. The
organisms must still be active or moribund, but not dead before being fixed. Failure to
follow proper fixation procedures will interfere with the interpretation of lesions in
anatomic pathology studies.
As with other monitoring methods, samples must be accurately labeled at the time of
collection and routine QA/QC procedures should be instituted, including tracking of
samples, careful recording of methods used, using fresh solutions, and treating both
control and exposed samples equally. Whenever possible, organisms for each group to
be tested should be of the same species, age, and sex. Sections of tissues and organs,
or for small organisms, the whole animal, should also be prepared as uniformly as
possible with respect to homogeneity and orientation so that microscopic observations
can be made on the same organs and areas for each specimen. Subsamples of sections
should be examined (blind) by another pathologist to confirm the diagnoses. See U.S.
EPA (1987a) for additional information on QA/QC procedures for histopathological
examinations.
Statistical strategies may mitigate the high costs of pathobiological monitoring meth-
ods. As discussed earlier, power analyses considering the strategy of compositing
samples can often lead to a cost-effective monitoring design strategy. Power-cost
analyses are necessary in selecting the appropriate sample/replicate number, sample
location, and sampling frequency. If the primary objective of a monitoring program is
to determine pathobiological differences among sampling locations, composite sam-
pling may be an appropriate strategy. However, a limitation of composite sampling is
the inability to directly estimate the range and variance of the population of individual
' samples. Similarly, the use of space- or time-bulking strategies will severely limit a
monitoring program's ability to assess spatial and temporal heterogeneity of the
samples. See Statistical Design Considerations: Composite Sampling, Statistical
Power, and Power-Cost Analysis (Section B.3).
Given that the monitoring program must accommodate a fixed level of sampling cost,
the best strategy for pathobiological monitoring is to collect more replicates at fewer
locations. For histopathology, from 25 to 100 animals per species may need to be
surveyed to detect low incidences of disease. The selection of appropriate statistical
analyses must be made with these limitations in mind, as well as the specific types of
statistical analyses that can be performed on different types of data. For example,
quantitative information from measurements of enzyme levels or computerized image
analysis/morphometric programs may be effectively analyzed by parametric statistics
B9.5
Statistical Design
Considerations
-------
B-181
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(if the data fit the conditions for normal distributions, limitations of assumptions, etc.),
but histopathological observations may need to be rated on presence/absence or
categorized qualitative scales that will require non-parametric techniques. U.S. EPA
(1987a) provides additional information on the types of sampling designs and statisti-
cal tests that may be appropriate for these data.
B9.6
Use of Data
Data Interpretation
Data interpretation for pathobiological methods may be limited because there are
relatively few trained personnel, facilities and equipment may be expensive, and
references for techniques that are newly emerging may be scarce. However, because
of the recent interest in developing biomarkers for monitoring the effects of natural
and anthropogenic environmental stresses on aquatic organisms, many research
programs are underway at Federal, State, and academic facilities to develop, standard-
ize, and validate the most promising biomarkers for sentinel species that will establish
cause and effect links for pollutant exposure. Basic laboratory research and experi-
mental studies must be conducted in conjunction with field work to elucidate the
relationships between contaminant levels, structural, biochemical, or functional
pathologies, and population health (Johnson and Bergman, 1984; McCarthy, 1990). A
critical concern is the coordination of efforts and creation of a multidisciplinary
approach.
It is important to establish baseline data on selected species as well as to demonstrate
the alterations in the health of those species due to contaminant exposures. Methods
and techniques, terminology, and interpretation of lesions and effects must be stan-
dardized. In addition to a rapidly expanding body of literature on baseline measures of
health and histological atlases for several species of fish and shellfish, and courses,
workshops, meetings, and special symposia (e.g., Responses of Marine Organisms to
Pollutants/Woods Hole, MA and Plymouth, England; Annual Aquatic Toxicology
Workshop, Canada), several professional societies (e.g., Society for Invertebrate
Pathology, American Fisheries Society/Fish Health Section) are facilitating training in
techniques and communication among investigators and laboratories. Diseases offish
and shellfish have been reviewed by several authors (i.e., Sparks, 1985; Ferguson,
1989; Roberts, 1989; Sindermann, 1990). For histopathological studies of tumors in
fish, standardization of nomenclature will be available in Dawe et al. (in press).
As for any monitoring program, long-term studies of health and disease in aquatic
organisms will aid in identifying and interpreting observed pathobiological effects.
Furthermore, it will be important to archive data so that it will be available for future
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B-1821
(I i f ' ft
comparisons. The Registry of Tumors in Lower Animals at the National Museum of
Natural History, Smithsonian Institution, Washington, D.C. houses over 3500 cases of
neoplastic and nonneoplastic lesions representing a wide variety of host aquatic
species, pathogens, and environmental stresses from field and laboratory studies
conducted around the world. The Registry also contains the Registry of Marine
Pathology, originally developed by the National Marine Fisheries Service (NMFS),
and the collection of crustacean histopathological material researched by Dr. Phyllis T.
Johnson, NMFS. Although primarily serving as a clearinghouse for information on
neoplastic diseases and research, the Registry is also able to direct inquiries for infor-
mation on nonneoplastic diseases, and the materials archived there are available for
study by qualified investigators. Data collected from the long-term monitoring efforts
of NOAA's National Status and Trends Program (NS&T) and the EPA's Environmen-
tal Monitoring and Assessment Program (EMAP) will also provide useful information
for the interpretation of the various pathobiological methods that are being employed
in these studies.
Data Integration
Measuring or evaluating the effects of stressors on fish and shellfish by
pathobiological indicators is difficult primarily because of the large number of vari-
ables that can influence biological response. Variables including water temperature,
nutritional status, species, sex, reproductive and developmental stages, and physiologi-
cal functions can render tests difficult to compare and evaluate.
Anderson (1990a) outlined a plan including four levels of study in a tiered approach to
investigate the effects of stress on the immune system and to quantify the possible
contribution of these environmental and physiological variables on the stress response.
The plan, generally speaking, appears to be applicable to all pathobiological methods
and may be adapted to them using the following four levels: (1) observations of fish
and shellfish populations in the field; (2) studies of caged fish or shellfish in the field;
(3) in vivo exposures in the laboratory; and (4) in vitro assays in the laboratory.
The first level of study includes the collection of information from field observations
on relative abundance, reduction in sport-fish catches, or decline in commercial
harvests. Reduced yields reported by anglers in previously productive fishing grounds
may be correlated with environmental stressors or pollutants occurring at these sites.
If sick or moribund fish or shellfish are called to the attention of local biologists by
anglers, gross morphological descriptions might be made and isolation of the disease
agent attempted.
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' /
The second level of investigation involves the use of caged fish or shellfish placed in
the field to test for the presence or action of environmental stressors. Groups of fish or
shellfish can be placed in pens or cages at suspected sites and their responses (signs of
disease, acute mortality) can be compared with those of control fish or shellfish caged
in unstressful areas. Organisms from the cages can be sampled at various intervals and
the intensity of various immune responses quantified as a function of the time the fish
or shellfish were exposed. Levels of immune responses can be compared by injecting
specific disease agents into both control and test organisms, and recording disease and
death rates.
The third level of assay is in vivo laboratory tests where immune response can be
evaluated in experiments with calibrated dilutions of specific contaminants and kinetic
measurements of the immune response at each dilution. Challenges with disease
agents can be more easily controlled in the laboratory to provide information on how
the stressor makes the fish or shellfish more susceptible to specific pathogens.
A fourth level of investigation is the recently developed method for testing the effects
of chemicals, drags, and other stressors in vitro. Spleens and other immunopoietic
organs can be removed from fish and placed in tissue culture media and their reactions
to pollutants (Anderson et al., 1986) tested using the passive hemolytic plaque assay.
Use of this method to measure the effects of stressors allows maximal control of the
levels of pollutants. Because the immune response is monitored under defined labora-
tory and environmental conditions, important information is obtained about how
specific stressors affect the immune response. In vitro methods require fewer fish than
in vivo methods because tissue and organ samples can be divided into many sections,
which also reduces the variability of responses.
McCarthy (1990) presented a research strategy to validate biomarkers and provide the
scientific understanding necessary to interpret biomarker responses. An evolving
monitoring program was proposed that focused broadly on evaluation of contamina-
tion in an array of ecosystem types. The challenges and obstacles to be addressed in
such a program include the following:
• The quantitative and qualitative relationships between chemical exposure,
biomarker response, and adverse effects must be established
• Responses due to chemical exposure must be distinguishable from natural
sources of variability (e.g., ecological and physiological variables, species-
specific differences, and individual variability) if biomarkers are to be
useful in evaluating contamination
-------
B-184 ]
• The validity of extrapolating between biomarker responses measured in
individual organisms and some higher-level effect at a population or
community level must be established
• The use of exposure biomarkers in animal surrogates to evaluate the
potential for human exposure should be explored
The plan is an ambitious multi-year research and development program involving the
formulation of a long-term, interagency, interdisciplinary activity such as the Environ-
mental Monitoring and Assessment Program (EMAP) by EPA in cooperation with
other federal agencies.
Rationale
• Pathobiological methods provide information concerning biological organ
systems that, through an evaluation of organ structure, activity, and func-
tion, can be used to determine adverse effects of pollutants in the environ-
ment.
B9.7
Summary and
Recommendations
• Pathobiological methods should be used in concert so that cause and effect
type relationships can be evaluated.
Monitoring Design Considerations
• Sampling stations should be located along a contamination gradient (i.e.,
from highly contaminated to uncontaminated). This type of sampling
strategy will allow dose-response relationships to be evaluated.
• Fish pathobiological monitoring should be conducted only if the target
species could reasonably be expected to have spent a considerable amount
of time within the area of contamination.
• Large sample sizes will frequently be required for fish pathobiological
monitoring due to natural variability among individuals and taxa.
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B-185
Existing Analytical Methods
Anatomic Pathology Methods-
• Tissues are examined with the naked eye or the aid of a microscope
• Light microscopic (LM) methods
- detect changes at the cellular level
- organ and time-series specific
• Electron microscopic (EM) methods
- detect subcellular changes
- specific as to area of cell or site of pollutant action
• Histochemical methods
- detect enzyme systems or reaction products in the cell
- specific to chemical class
Reproduction/Development Methods-
• Reproductive capacity is reflected in population recruitment and abundance
• Methods that examine the egg itself provide a direct measure of reproduc-
tive effects from pollutants
• Cytogenetic (DNA) tests
- in vitro tests offer greatest flexibility in terms of sensitivity,
expense, and time required, and are frequently conducted for
convenience and availability
- sister-chromatid exchange (SCE) assays are DNA damage tests
used as a screening tool, and are dose-responsive and sensitive to
low concentrations of pollutants
- the aneuploidy technique is simple and easy to use, but is inaccu-
rate for sublethal effects
- chromosomal alteration analysis
Biochemical Methods-
• Several methods are specific for a certain compound class
-------
B-186 I
• Use a suite of indicators for environmental monitoring
• Microsomal mono-oxygenase (MO) assays could serve as sensitive bio-
logical indicators for certain chemical classes
• Metallothionein (MT) assays are nonspecific for metal exposure, but can
provide information on the likelihood of a particular metal pool producing
a pathological effect
• Stress proteins are not pollutant-specific, but could result from generalized
stress
Immunologic Methods—
• Immunologic assays can provide information on pollutant-induced stress
effects; however, they may require confirmatory assays
• Many animals should be sampled because of natural variability among
animals
Physiologic Methods-
• Serious "interferences" can be caused by stress induced during collection
and may limit the potential usefulness of physiological tests because effects
of toxicants cannot be distinguished from those induced during handling of
the wild organisms
• Hcmatologic methods for fish, such as measurements of hematocrit
(packed cell volume), hemoglobin concentration, erythrocyte count and
leucocyte count (or volume) can be successfully used, although the fish are
not immune to stress of capture, hematologic methods are influenced far
less than some other measurements such as blood glucose
• The leucocrit can be a more sensitive measure of metal pollution than the
hematocrit
QA/QC Considerations
• PathobiologScal methods have a wide range of sensitivities
-------
B-187
• Organisms must be carefully handled and properly prepared for each
method
• The main areas of concern with regard to analytical QA/QC are precision,
accuracy, representativeness, completeness, and comparability
Statistical Design Considerations
• Composite tissue sampling consists of mixing tissue samples from two or
more individuals collected at a particular location and time
• Space- (combining composites from several locations) and/or time- (com-
bining several composites over time from one location) bulking strategies
should be used judiciously since information concerning spatial and
temporal heterogeneity may be lost
• Power analyses have shown that for a fixed level of sampling effort, a
monitoring program's power is generally increased by collecting more
replicates at fewer locations
• The appropriate statistical tests must be performed and may vary depend-
ing on the type of data generated
Use of Data
• Data use and interpretation may be limited by relatively few trained
personnel and expensive equipment, but many research and training
programs are now underway at Federal, State, and academic facilities to
improve biomarker methods in sentinel species
• Basic laboratory research must be conducted and biological methods must
be tested in the field
Information communication
- need to coordinate efforts, communicate and create a
multidisciplinary approach to relate lab and field studies and
establish cause and effect type relationships
-------
B-1881
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Data integration
long-range research strategies should be followed to validate
bioraarkers and provide scientific understanding necessary to
interpret biomarker responses
Adams, S.M. (ed.) 1990. Biological Indicators of Stress in Fish. American Fisheries B9.8
Society Special Symposium No. 8. Literature Cited
and References
Adams, S.M., L.R. Shugart, and G.R. Southworth. 1990. Application of bioindicators
in assessing the health of fish populations experiencing contaminant stress. In: Bio-
markers of Environmental Contamination (McCarthy, J. and L. Shugart, eds.), pages
333-353. Boca Raton, FL: CRC Press.
Anderson, D., B. Roberson, and O. Dixon. 1979. Cellular immune response in
Rainbow Trout, Salmo gairdneri Richardson to Yersinia Ruckeri O-antigen monitored
by the passive haemolytic plaque assay test. J. Fish Dis. 2:169-178.
Anderson, D., O. Dixon, and E. Lizzio. 1986. Immunization and culture of Rainbow
Trout organ sections in vitro. Veterinary Immunology and Immunopathology
12:203-211.
Anderson, D. 1990a. Immunological indicators: Effects of environmental stress on
immune protection and disease outbreaks. American Fisheries Society Symposium
8:38-50, Washington, DC.
Anderson, D. 1990b. Passive hemolytic plaque assay for detecting antibody-produc-
ing cells in fish. In: Techniques in Fish Immunology. (Stolen, J., J. Fletcher, D.
Anderson, B. Roberson, W. van Muiswinkel, eds). Fair Haven, NJ: SOS Publications.
Anderson, D., O. Dixon, and W. van Muiswinkel. 1990. Reduction in the numbers of
antibody-producing cells in Rainbow Trout, Oncorhynchus mykiss, exposed to suble-
thal doses of phenol before bath immunization. In: Aquatic Toxicology. (Nriagu, J.,
cd.). A Wiley-Interscience Publication, John Wiley and Sons.
Anderson, R.S. 1987. Immunocompetence in invertebrates. In: Pollutant Studies in
Marine Animals (Giam. C.S. and L.E. Ray, eds.). Boca Raton, FL: CRC Press.
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.. •/•
Anderson, R.S. 1990. Effects of pollutant exposure on bactericidal activity of
Mercenariamercenariahemolymph. In: Biological Markers of Environmental
Contaminants (McCarthy, J.F. and L.R. Shugart, organizers), American Chemical
Society, Los Angeles, CA, 25-30 September 1990.
Blaxhall, P. and K. Daisley. 1973. Routine haematological methods for use with fish
blood. J. Fish Biol. 5:771.
Bouck, G.R. 1984. Physiological responses offish: problems and progress toward
use in environmental monitoring. In: Contaminant Effects on Fisheries (Cairns, V.W.,
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B-1901
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B10.0 Bioaccumulation
Bioaccumulation is the overall process of biological uptake and retention of chemical
contaminants obtained from foods, water, sediments, or any combination of exposure
pathways - a list of terms may be found in Table B10-1. It is a consequence of an
organism's physiological limitations to transform and excrete the invading chemical
substances.
TABLE B10-1. LIST OF TERMS
= the fraction of the total contaminant in tho surrounding
medium which is correlated with a quantitative biological
response, such as bioaccmmulatioo
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B-2001
column at very low concentrations which are difficult to detect, yet can be accumu-
lated in detectable amounts in fish and shellfish tissue. In those cases where toxics can
be measured in the water and sediments, there is a large degree of uncertainty associ-
ated with even the most sophisticated models for predicting uptake and bioaccumula-
tion. Direct monitoring of the concentration of contaminants of concern in the tissue
of selected estuarine organisms can provide a spatial and temporal record of contami-
nant concentrations and bioaccumulation in the estuary.
Monitoring of bioaccumulation in the estuary will provide the data which links expo-
sure and effects, and thus can generate important insights into ecological effects,
human health risks, and routes and extent of pollutant exposure. This information can
be used to both evaluate the effectiveness of the CCMP in mitigating the potential
effects due to bioaccumulation and to provide an early warning system for the poten-
tial risks associated with the bioaccumulation of xenobiotics.
Bioaccumulation monitoring studies have varied substantially, depending upon study-
specific objectives. Generally, bioaccumulation monitoring programs whose objective
was to assess human health effects were significantly different in their selection of
target species and tissues when compared to those which were intended to assess
impacts to estuarine communities. Human health risk assessments usually examine the
muscle tissue of commercially important fish and shellfish, whereas environmental
impact assessments often survey organisms at lower trophic levels (e.g., infaunal
macroinvertebrates). Unfortunately, formulations which would allow comparisons of
bioaccumulation data between different species and/or different tissues types have not
yet been developed. Thus, it is essential that monitoring design elements be standard-
ized to allow for comparisons among estuarine studies.
A common deficiency in many programs, is the inability to collect sufficient tissue
biomass of appropriate species across sampling locations throughout the study. The
selection of appropriate species and tissues must account for natural fluctuations in
populations as well as changes due to anthropogenic perturbations. Indigenous species
initially present may not be available later, limiting temporal and spatial comparisons.
Selection of Target Species
A key component of any bioaccumulation monitoring program is the selection of
target species. Concentrations of chemical residues in tissues of target species serve as
indicators of contamination throughout the biological system. The fundamental
B10.2
Monitoring Design
Considerations
-------
B-201
,"•*
ss.., s. ss s
criterion is the ability to use the selected species to make comparisons between sam-
pling locations and sampling periods.
Target Species Characteristics - It is recommended that the target species possess
the following characteristics (U.S. EPA, 1985a):
• high bioaccumulation potential for selected contaminants of concern
• metabolic regulation of selected contaminants should be weak or absent in
order to assess a "worst case scenario"
• abundant, temporally and spatially, to allow for adequate sampling
• large enough to provide adequate amounts of tissue for analysis
• sessile or sedentary in nature to assure bioaccumulation is representative of
the study area
• easily collected
• existing database of exposures and sensitivity
The EPA has provided a list of suggested fish and macroinvertebrate target species
(Tables B10-2 and B10-3; U.S. EPA, 1985a) for different regions in the United States.
Secondary considerations, based on economic importance and status as a bioassay
organism, may be applied to further winnow the list of candidate target species to a
practical number of species to be analyzed.
Candidate target species should be objectively ranked based on ecological characteris-
tics which would enhance their potential for bioaccumulation, and facilitate sampling
and/or analytical procedures (U.S. EPA, 1985a). Macroinfauna have several advan-
tages as target species, including:
• are sedentary; therefore, bioaccumulation will be representative of the
study area
• represent a significant food source for higher trophic levels and therefore
may indicate contaminants available in the food chain
-------
B-2021
TABLE B10-2. HIGHEST RANKING CANDIDATE FISHES FOR
USE
AS BIOACCUMULATION MONITORING SPECIES
State Locality
MASSACHUSETTS Swampscolt
Lynn
South Essex
Boston
Fall River
New Bedford
RHODE ISLAND Newport
NEW YORK Upper East River
Lower East River
Lower Hudson River
NEW JERSEY Cape May
VIRGINIA Portsmouth
Virginia Beach
Species
Winter flounder
Yellowtail flounder
Ocean pout
Windowpane
Winter flounder
Yellowtail flounder
Ocean pout
Winter flounder
Yellowtail flounder
Windowpane
American eel
Ocean pout
Winter flounder
Yellowtail flounder
Ocean pout
Windowpane
Winter flounder
Windowpane
Winter flounder
Scup
Summer flounder
Winter flounder
Scup
Weakfish
Winter flounder
Windowpane
Weakfish
Spot
Scup
American eel
Hogchoker
Spot
Red hake
Windowpane
Summer flounder
Spot
Summer flounder
Atlantic croaker
Hogchoker
Spot
Red hake
Summer flounder
Secondary Selection
Criteria
Economic Bioassay
Importance Species
Yes
Yes
No
No
Yes
Yes
No
Yes
Yes
No
No
No
Yes
Yes
No
No
Yes
No
Yes
Yes
Yes
Yes
Yes
Yes
Yes
No
Yes
Yes
Yes
No
No
Yes
No
No
Yes
Yes
Yes
Yes
No
Yes
No
Yes
Yes
Yes
No
No
Yes
No
No
Yes
No
No
No
No
Yes
No
No
No
Yes
No
Yes
No
Yes
Yes
No
No
Yes
No
No
Yes
No
No
No
Yes
No
No
Yes
Yes
Yes
No
No
Yes
No
Yes
-------
B-203
State
CALIFORNIA
(NORTHERN)
CALIFORNIA
(SOUTHERN)
WASHINGTON
Locality
San Francisco
Oakland
Monterey
Santa Cruz
Watsonville
Goleta
Santa Barbara
L.A. County
Orange County
Hyperion
Oceanside
Escondido
San Elijo
San Diego
Central Puget Sound
TABLE B10-2.
(continued)
Species
English sole
Pacific sanddab
Big skate
English sole
Starry flounder
Pacific staghom sculpin
English sole
Curlfin sole
English sole
English sole
Curlfin sole
Dover sole
Pacific sanddab
Longspine combfish
Spotted cusk-ecl
English sole
Pacific sanddab
Dover sole
Curlfin sole
English sole
Dover sole
Pacific sanddab
English sole
Dover sole
Longspine combfish
Big skate
California skate
Dover sole
Blackbelly eelpout
Pacific sanddab
English sole
Dover sole
English sole
Pacific sanddab
Longspine combfish
English sole
Dover sole
Rock sole
Spotted ratfish
Rex sole
C-O sole
Secondary Selection Criteria
Economic
Importance
Yes
Yes
No
Yes
Yes
No
Yes
Yes
Yes
Yes
Yes
Yes
Yes
No
No
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
No
No
No
Yes
No
Yes
Yes
Yes
Yes
Yes
No
Yes
Yes
Yes
No
Yes
Yes
Bioassay
Species
Yes
No
No
Yes
No
No
Yes
No
Yes
Yes
No
No
No
No
No
Yes
No
No
No
Yes
Yes
Yes
Yes
No
No
No
No
No
No
No
Yes
No
Yes
No
No
Yes
No
No
No
No
No
-------
B-2041
TABLE BjO:3. , „„
RECOMMENDED LARGE MACROINVERTEBRATE SPECIES
FOR BIOACCUMULATION MONITORING
Region
Recommended Species3
Massachusetts to Virginia
Alaska to California
Florida, Virgin Islands, and
Puerto Rico
Hawaii
-American lobster (ffomarus americantis)
Eastern rock crab (Cancer irroratus)
Hard clam (Afercenaria mercenaris)
Soft-shell clam (Mya arenaris)
Ocean quahog (Arttea isfandica)
Surf clam. (Spisiila solidissima)
.'Edible mussel (Mytilus edulis}
Spiny \QbS(&tj(Panulirus mtermptus}
Dungeness crab (Cancer magister)
Rode crab (Cancer antennarius)
Yellow crab (Cancer anthonyfy
Red crab (Cancer prodiictus}
California mussel (Mytilus calif
Spiny lobster (Panulirtts argus)
, Spiny lobster (PanulirUs penicittatu$y
* Addiu'onal species that may occur at specific discharge sites and are considered
acceptable bioaccumulation monitoring Species include the, American oyster
(Crassostrea virginica) and the Pacific oyster (Crassastrea gigas).
However, macroinvertebrates may not provide sufficient tissue biomass for analyses.
Adult fish of the species listed in Table B 10.2 provide sufficient tissue biomass for
analyses and are direct measures of contaminants available to human populations. How-
ever, fish are usually motile and bioaccumulation measured may not be representative of
the study area.
It is recommended that bioaccumulation studies be conducted for a number of species.
-------
B-205
Species have different bioaccumulation potentials for various contaminants; monitor-
ing multiple species will ensure that bioaccumulation of a number of contaminants are
sufficiently evaluated. For example, oysters and other bivalves are ideal for monitor-
ing bioaccumulation of PAH's due to their limited ability to metabolically transform
them. Monitoring of species along a food chain might provide information concerning
the transfer of contaminants to higher trophic levels. Species with extensive historical
bioaccumulation data are also preferred. Where possible, species being collected for
other aspects of the monitoring program (e.g., fisheries data, pathobiology, pathogens)
should be included in the bioaccumulation analyses, to enhance the coordinated
evaluation of data.
Caged Indicator vs. Indigenous Species - The California Mussel Watch Program, the
U.S. Mussel Watch Program, and the National Oceanic and Atmospheric Administra-
tion (NOAA) Status and Trends Program have employed the use of both resident and
caged transplant mussels to monitor bioaccumulation of toxic chemicals over space
and time (Goldberg et al., 1978; Boehm, 1984; Ladd et al, 1984). Caged indicator
species offer several advantages over indigenous species:
• the biology and ecology of these indicator species are usually well de-
scribed
• the descriptions of culture and/or maintenance of the organisms under
laboratory conditions are often available
• the method provides control of initial temporal and spatial variation of
individuals and/or biomass
• the method allows the use of a specific age, size and/or genetic stocks
• the method assures on-site bioaccumulation
By controlling for initial conditions, the rate of change of tissue contamination may be
calculated. In addition, the Long Island Sound Estuary Program has shown that
chemical residue analyses on caged indicator species appears to be a promising
approach for identifying sources of pollution (U.S. EPA, 1982). However, transfer
through the water is the only exposure pathway assessed; adequate methods for caging
sediment ingesting organisms are not yet available. Because bioaccumulation of
sediment-sorbed contaminants is probably mediated by transfer through interstitial
waters (Knezovich and Harrison, 1987) and results obtained often do not relate to
species found on site, caged organisms may not provide an accurate estimate of the
bioavailability of certain contaminants occurring at the site.
-------
B-2061
The advantages of indigenous species are:
• results obtained will relate directly to those species which may be impacted
and provide a direct measure of potential risks to human health
• no cost of transplanting organisms
• no possibility of loss of cage/buoy system
• no possibility of introduction of a "nuisance" species
However, common indigenous species may not meet the criteria for use as bioaccumu-
lation target species, negating the advantage of using a native species. The most
common problem is collecting sufficient tissue biomass of appropriate indigenous
species across sampling locations throughout the study.
Selection of Tissues
The type and location of tissue analyzed will depend upon the objectives of individual
monitoring programs. It is essential that all analyses be conducted with the same
tissue type from the same species in order that scientifically and statistically valid
comparisons may be made.
Target Tissues— In fish, liver tissue is closely associated with regulation and storage
of many toxic substances (Fowler, 1982). Its high fatty tissue content tends to accu-
mulate hydrophobic contaminants. Thus, contaminant levels in the liver can be used to
estimate the range of contaminants being assimilated. For macroinvertebrates, hepato-
pancreas or digestive gland tissue performs functions analogous to fish liver tissue.
Contaminants in edible muscle tissue represent those contaminants that are retained in
a form that allows transfer to humans. Sampling of muscle tissue is appropriate for
human exposure assessments and quantitative health risk determinations. Within a
fillet, contaminant concentrations may vary, therefore it is recommended that a consis-
tent location within the muscle tissue be analyzed (U.S. EPA, 1989).
Whole body analyses ought to be conducted when predators consume the whole body
of the target organism. If organisms are not cleansed of materials contained in the
digestive tract, contaminants in the gut contents will be included in the analyses. To
provide the most accurate estimate of the total amount of contaminants available to
most macroinvertebrate predators, this type of depuration may not be required.
-------
« < A -s
s ""•"•••.
ss •y.s.w, tf, -. v
B-207
Sediment Total Organic Carbon and Acid Volatile Sulfides, and Tissue Lipid
Normalization - Toxic sediment concentrations of hydrophobic contaminants have
been found to be related to the total organic carbon content (TOC) of the sediment
(Karickhoff et al., 1979). The bioavailability of some metal contaminants has been
found to be related to the acid volatile sulfide (AVS) concentrations of the sediment
(DiToro et al., in press; see Section B4.2). TOC and AVS normalizations have been
conducted to estimate the concentrations of sediment contaminants which are bioavail-
able over different sampling locations. Sediment TOC and AVS normalizations are
recommended to account for the variability in bioavailable sediment contaminant
concentrations between locations.
Lipids appear to be a storage site of organochlorines, hydrocarbons, and other hydro-
phobic contaminants in a variety of organisms (de Boer, 1988). In fact, just as TOC
and AVS are considered by many to be important parameters in defining organic and
metal concentrations in sediments, lipid content is considered the salient parameter in
defining hydrophobic residue concentrations found in tissues (Phillips and Segar,
1986). Likewise, tissue lipid normalizations have been suggested to account for the
variability in tissue contaminant concentrations between individuals.
Normalization of sediment contaminant concentrations to TOC, AVS, and tissue
contaminant concentrations to the tissue lipid concentrations have been made to permit
comparisons of toxic chemical residues in tissues between studies, locations, individu-
als, and tissue types. These sediment and tissue normalizations assume the following:
• contaminants partition predominantly to sediment TOC, sediment AVS,
and organismal lipids
• contaminants partition reversibly between the sediment particles and
organism
• rapid steady-state kinetics of contaminants
• sediment is the only source for the bioaccumulation
Lipid normalization has been performed in order that individuals with differing body
fat levels may be compared. Again, it is essential that all analyses be conducted with
the same tissue type from the same species in order to ensure that scientifically and
statistically valid comparisons may be made. TOC/lipid normalized accumulation
factors (AF) have also been used to predict tissue residue concentrations (Ferraro
etal, 1990; Lake etal., 1987).
-------
y> st s
/•*"
Three-fold differences in lipid concentration may result as a consequence of various
lipid analysis techniques. Development of protocol standardization or intercalibration
between lipid techniques are required before chemical residues in tissues can be
compared among studies. Microtechniques have.been developed for analyses of lipids
from single individuals (Gardner et ol,, 1986).
Finally, the decision to normalize for TOG, AVS, and percent lipid will depend upon
monitoring program objectives. For example, if the monitoring objective is to identify
"hot spots" - i.e., those areas where high body burdens present a risk to human and
ecological health - normalization may not be appropriate. Whereas, if the objective is
to identify temporal trends in bioaccumulation rates, normalization may be justified.
Selection of Sampling Period
The timing of sampling should be based on biological cycles which influence an
organism's susceptibility to bioaccumulation. The frequency of sampling should be
related to the expected rate of change in tissue concentrations of contaminants. A
consistent sampling period is recommended in order that spatial and temporal compari-
sons may be conducted.
For crustaceans, just after molting, before hardening of the integument occurs restrict-
ing its permeability, there is a significant increase in potential for bioaccumulation of
toxic contaminants.
For many aquatic vertebrate and invertebrate organisms, the reproductive cycle exerts
a major influence on tissue concentrations of many contaminants - especially hydro-
phobic compounds (Phillips, 1980). For many species, lipophilic (hydrophobic)
contaminants arc transferred from the muscle and liver to the eggs as lipids are mobi-
lized and transported to the egg during oogenesis (Spies et al., 1988; Gardner et al.,
1985). There is evidence that lipophilic contaminants have deleterious effects on the
developing egg and/or the larvae (Spies et al, 1988; Hansen et al., 1985; Niimi, 1983).
Transfer of contaminants from adult to egg, and its potential impacts to future fisheries
recruitment and fisheries production are under investigation.
It is recommended that target species be sampled when tissue contaminant concentrations
are expected to be at their highest level in order to evaluate worst-case scenarios. For
those organisms where the body is consumed whole, contaminant levels are usually at
their highest at or just before spawning. For organisms where the muscle tissue is con-
sumed, contaminants in muscle tissue usually reach a peak well before spawning.
-------
B-209
B10.3 Questions to be considered during the choice of an appropriate analytical method
Existing include the parameters of interest, desired detection limits, sample size requirements or
Analytical restrictions, methods of preservation, technical and practical holding times, and matrix
Methods interferences. Several U.S. EPA documents (1986a, 1987a, 1990b) discuss the com-
mon analytical problems encountered during monitoring analyses of tissue samples.
Chemical Residue Analyses
Several factors determine achievable detection limits for a specific contaminant,
regardless of analytical procedure - a list of analytical procedures and EPA method
numbers is given in Table B10-4. These factors include:
• size of the tissue available for analysis - in general, the ability to detect
low contaminant levels is improved with greater amounts of tissue avail-
able for analysis; a minimum of 30 g (wet weight) is usually considered
adequate (U.S. EPA, 1987a)
• presence of interfering substances
• range of pollutants to be analyzed - an optimal method for a specific
pollutant may be developed without regard to potential effects on other
parameters
• level of confirmation - qualitative (e.g., presence or absence) or quantita-
tive (e.g., residue concentrations) analyses
• level of pollutant found in the field and in analytical blanks
Selection of appropriate methods ought to be based on a trade-off between full-scan
analyses, which are economical but cannot provide optimal sensitivity for some
compounds, and alternate methods that are more sensitive for specific compounds but
can result in higher analytical costs.
Metals and Metalloids- Nitric acid/perchloric acid digestions should be considered
for analysis of tissue samples (U.S. EPA, 1987a). Perchloric acid is especially useful
for the dissolution of fats.
Trace element analyses by ICP (U.S. EPA Method 6010) allows for several elements
to be measured simultaneously. Detection limits of ICP for most metals are generally
-------
B-210]
"••Mv.
comparable to those achieved by graphite furnace AAS (GFAAS); however, detection
limits for several metals are significantly lower using AAS: arsenic, selenium, and
mercury.
GFAAS is the recommended analytical method for detection of arsenic, selenium, lead
and thallium in tissues (U.S. EPA, 1987a). Cold vapor AAS (U.S. EPA method 7470)
analysis is the only recommended technique for mercury (U.S. EPA, 1986a).
GFAAS is more sensitive (i.e., lower detection limits) than flame AAS, but is more
subject to matrix and spectral influences - GFAAS requires particular caution with regard
to laboratory contamination. GFAAS requires more skilled laboratory technicians. Both
AAS methods require that the concentration of each element be determined by a separate
analysis, making the analysis of a large number of contaminant metals both labor-intensive
and relatively expensive compared to ICP. ICP is therefore the preferred method of
analysis for metals, except arsenic, selenium, lead, mercury and thallium.
Semi-Volatile Organic Compounds - Analysis of semi-volatile organic compounds
involves a solvent extraction of the sample, cleanup of the characteristically complex
extract, GC analysis, and quantification (U.S. EPA, 1987a; U.S. EPA, 1986a), There
arc two GC/MS options for detecting extractable organic compounds: internal stan-
dard technique and isotope dilution. The isotope dilution technique is recommended
because reliable recovery corrections can be made for each analyte with a labeled
analog or a chemically similar analog (U.S. EPA, 1986a). The isotope dilution method
is more expensive and less widely employed than the internal standard technique,
which is the current method of choice in the Contract Laboratory Program (CLP).
Mass spectrometry provides positive compound identification by comparison of both
retention time and spectral patterns with standard compounds.
The identification of pesticides and PCB congeners can be made by GC/ECD analysis
(U.S. EPA Method 8080). GC/ECD provides greater sensitivity relative to GC/MS,
however GC/ECD does not provide positive compound identification. Confirmation of
pesticides and PCBs on an alternative GC/ECD or preferably by GC/MS, when sufficient
concentrations occur, is recommended for reliable results (U.S. EPA, 1985b). All other
organic compound groups are recommended for analysis by GC/MS.
Volatile Organic Compounds- Analysis of volatile organic compounds also in-
volves GC analysis and quantification (U.S. EPA, 1987a; U.S. EPA, 1986a). The
purge and trap GC/MS technique is employed for detecting volatile organic com-
pounds in water.
-------
B-211
TABLE B10-4. LIST OF EXISTING ANALYTICAL TECHNIQUES
-------
B-2121
for chemical analyses require all sampling equipment - e.g., siphon hoses, scoops,
containers—be made of noncontaminating material and be cleaned appropriately prior
to use. Efforts should be made to minimize handling and to avoid sources of contami-
nation including potential airborne contamination (e.g., stack gases, cigarette smoke).
To avoid contamination from ice, whole samples should be placed in watertight plastic
bags, and immediately cooled in a covered ice chest. Only if the samples will not be
analyzed for metals, they may be wrapped in acid-rinsed aluminum foil with the shiny
side out before being placed in plastic bags. Furthermore, samples and sampling
containers should not be touched with ungloved fingers.
Sample Handling and Storage
For analyses of metals, samples should be frozen and kept at -20°C (Table B10-5).
Although specific tissue sampling holding times have not been recommended by the
EPA, a maximum of 6 months (8 days for mercury; ASTM, 1991) would be consistent
TABLE B1Q-5.
SAMPLE PRESERVATION AND STORAGE PARAMETERS
Sample,
Storage
Analyte
Container3 'Size
Preservative Lifetime
All metals (except Hg)
Hg
P,G r
Freezed " ', 71,6 mo®
Freeze*! s ,,,,£daysc
a P s linear polyethylene, G =* boroslllcate glass, TFB «tefrafluoroethylene*
b Wet weight s"s ^ ,"•.*"
"" \ s' "" "•
c Suggested. No EPA criteria' exist. J|g holding time 8 days, " " -
%? ..-MX:''
d Post-dissection. ... -^
e If aliquot for Hg (aken from this 1 liter sample, cannot use linear jroiyethylene.
t * %
f Weight Is a minimum for one $ainple.%\StudieS using ;$pecificsorgans may requite
more. . " >' : . ..
-------
B-213
with that for water samples. For analyses of volatile compounds, samples should be
stored in the dark at 4°C (Table B10-6; U.S. EPA, 1987a). Analyses of volatile
compounds should be performed within 14 days of collection as recommended by U.S.
EPA (1990c). If analyses of semivolatile compounds will not be performed within the
recommended 7-day holding time, freezing of the samples at -20°C is advised. Hold-
ing times for frozen samples have not been established by the U.S. EPA. A general
guideline of a maximum of 6 months would be consistent with that for water samples
(U.S. EPA, 1987a).
Resectioning tissue samples for chemical analyses should only be conducted with
noncontaminating tools under "clean room" conditions. It is recommended that whole
tissue samples not be frozen prior to resection if analyses will be conducted only on
selected tissues because freezing may cause internal organs to rupture and contaminate
other tissues. A separate set of utensils for removing outer tissue and for resectioning
tissue for analysis is highly recommended. Incision troughs are subject to contamina-
tion and should not be included in the analysis.
TABLE B10-6,
SUMMARY OF SAMPLE COLLECTION AND PREPARATION
QA/QC REQUIREMENTS FOR ORGANIC COMPOUNDS
Maximum
Variable Sample Size3 Container1* Preservation Holding Time
Tissues (After Resection)
Semivolatiles 25 g
Votaries 5 g
<3,T
Freeze
Freeze
$ months0
^ Recommended field sample sizes for one laboratory analysis, If additional
laboratory analyses are required (ie., replicates). The field sample size should be
adjusted accordingly. ' "T,
~b G = Glass, A=Wrapped in altimihwn foil, placed in watertight plastic bags,
T«3?TFE (Teflon). - „-
c This is a suggested holding time. No IXS. EPA criteria exist for the preservatiGir~ -"
of this variable. - - , ,,„..,
-------
B-214 I
Laboratory Analyses
Laboratory performance as measured by method QA/QC protocols should be used to
evaluate and select appropriate analytical methods. Changes to laboratory protocols
should only be considered if new protocols meet established performance criteria.
Suggested QA/QC protocols and analytical performance criteria are discussed below.
QA/QC reports should describe the results of quantitative QA/QC analyses, as well as
other elements critical to the laboratory analyses of water, sediments, and tissues to
ensure proper interpretation of the results. It is recommended that these reports be
recorded and stored in a database for future reference.
Field QA/QC Checks - Travel blanks will indicate whether any contamination was
introduced by reagents in the field or during shipping of samples. Rinsate blanks are
designed to verify the absence of contamination carried over from one sample to
another due to inadequate cleaning of field equipment.
Held splits treated and identified as separate samples, may be sent to the same labora-
tory for analysis or one sample sent to a "reference" laboratory for comparison. In
addition, standard reference material may also be placed in a sample container at the
time of collection and sent "blind" to the laboratory.
Instrument QA/QC Checks - Calibration standards should be analyzed at the begin-
ning of sample analysis, and should be verified at the end of each 12-hr shift during
which analyses are performed (U.S. EPA, 1987b). The concentrations of calibration
standards should bracket the expected sample concentrations, sample dilutions or
sample handling modifications (i.e., reduced sample size).
Method QA/QC Checks- Analysis of method blanks should be conducted to demon-
strate the absence of contamination from sampling or sample handling in the labora-
tory. At least one method blank must be included with each batch of samples and
should constitute at least 5% of all samples analyzed.
Spike recovery analyses are required to assess method performance for the particular
sample matrix. Spike recoveries serve as an indication of analytical accuracy, whereas
analysis of standard reference materials measure extraction efficiency. Recommended
control limits include 75-125 percent recovery for spikes, and 80-120 percent recovery
for the analysis of standard reference materials.
-------
B-215
Replicates must be analyzed to monitor the precision of laboratory analyses. A
minimum of 5% of the analyses should be laboratory replicates. The control limits are
±20 percent relative percent difference for duplicates. Triplicates should be analyzed
on one of every 20 samples or on one sample per batch if less than 20 samples are
analyzed.
Tables B10-7 and B10-8 provide a brief summary of QA/QC for laboratory analyses.
A discussion of detection limits may be found in QA/QC Considerations in the Water
Column Chemistry chapter (Section B2.4).
B10.5 Statistical strategies may mitigate the high costs of collecting sufficient tissue biomass.
Statistical Design See also Statistical Design Considerations: Composite Sampling, Statistical Power,
Considerations and Power-Cost Analysis (Section B.3).
B10.6 Results of the bioaccumulation analyses can be used to establish spatial and temporal
Use Of Data trends in the bioaccumulation of toxicants of selected estuarine fish and
macroinvertebrates. Geographic maps of these trends would provide a survey of
trends throughout the estuary.
In addition, monitoring studies might identify existing and potential problem areas for
fish and macroinvertebrate contamination. Human health risk, as well as environmen-
tal risk assessment calculations may also be determined from data collected from these
studies. The information may be used to develop management strategies for commer-
cial and recreational fishing.
B10.7
Summary and
Recommendations
Rationale
Presence of toxics in waters and sediments of estuaries can have adverse
ecological and human health effects
Monitoring the accumulation of chemical residues in tissues of estuarine
organisms will provide information essential in relating the presence of
selected contaminants in estuarine waters and sediment to its transfer and
accumulation in estuarine organisms
-------
B-216]
VAC*,
TABLE B10-7.
SUMMARY OF QUALITY CONTROL SAMPLE
Analysis Type
Recommended Frequency of Analysis*
Surrogate spikes
Method blank
Standard reference
materials
Matrix spikes
Spiked method blanks
Analytical replicates
Held replicates
Required in every sample - minimum 3 neutral, 2 acid Spikes,
plus 1 spike for pesticide/PCB analyses, arid'3 spikes for
volatiles. Isotope dilution techniques (i.e., with all available
labeled surrogates) is recommended for full scan analyses and
to enable recovery corrections to be applied to data.
One per extraction batch (semi volatile organics) One per
extraction or one per 12 hour shift, which ever is most frequent
(volatileorganics) , "'" ,~ " ;'/!
<50 Samples: one per set of samples submitted, to lab
>50 samples; one per'SQ samples analyzed
Npi required if complete isotope dilution technique used "
<20 samples; one per set of samples submitted to lab
£20 samples: 5 percent of total! number of samples.
As many as required to establish confidence in method before
analysis of samples (i.e., when using a method for the first time
or after any method modification), "' - ', £'
<20 samples: one per set of .samples submitted to lab
5:20 samples: one triplicate and additional duplicates 'fora
minimum of 5 percent total replication,
•,*, s ••
At the discretion of the project coordinator.
a Frequencies listed are minimums: some programs may require higher levels of effort.
-------
wrtVASWW.
B-217
x ,,
TABLE B10-&
SUMMARY OF WARNING
AND CONTROL LIMITS FOR QUALITY CONTROL SAMPLE
Analysis Type
Recommended
Warning Limit
Recommended
Control Limit
Surrogate Spikes
Method Blank
Phthalate,
Acetone
Other Organic
Compounds
Standard
Reference Materials
Matrix Spikes
Spiked Method Blanks
Analytical Replicates
Field Replicates
Ongoing Calibration
10 percent recovery
30 percent of the analy te
1 jog total or 5 percent
oftheanalyte ,,, ,
confidence interval
(50-65 percent recovery)
(50-65 percent recovery)
{50 perceritirecovery)
5 yg total or 50 percent
oftheanalyte
2J5 }1§ total or 5 percent
of the analy te
95 percenfconfidence
interval for Certified
Reference Material
(50 percent recoveryy
(50 percent recovery)
±100 percent
coefficient o£ variation
25 percent 0L...
-------
B-218
ij«!,;:,as; a,,
• Monitoring program may provide information necessary to assess environ-
mental and human health risks associated with measured levels of bioaccu-
mulation
Monitoring Design Considerations
• It is recommended that consistent types of target species and tissues, data
normalization, and location and timing of sample collection be imple-
mented to allow for comparisons among studies
• Target species should possess the following characteristics:
- high bioaccumulation potential for selected contaminants of
concern
- metabolic regulation of selected contaminants should be weak or
absent
- abundant, temporally and spatially, to allow for adequate
sampling
- large enough to provide adequate amounts of tissue for analysis
- sessile, or sedentary in nature to assure bioaccumulation is
representative of the study area
- easily collected
• Analyses should be conducted on a number of different target species to
ensure sensitive measures of bioaccumulation for the contaminants of
concern
• Caged indicator species
- method allows for control of initial temporal and spatial variation
of individuals and/or biomass
- method allows for the use of specific age, size and/or genetic
stocks
Indigenous species
- results obtained will relate directly to those species which may be
impacted
- no possibility of loss of holding apparatus and test organisms
Target tissues
- fish liver or macroinvertebrate hepatopancreas analyses may be
used to estimate the range of contaminants being assimilated
-------
B-219
- muscle tissue analyses are appropriate for human exposure
assessments and quantitative health risk determinations
- whole body analyses ought to be conducted when predators
consume the whole body of the target organism
• TOC/AVS/Lipid Normalization
- recommend normalization to allow comparisons of chemical
residue concentrations in tissues between locations, individuals,
and tissue type - normalizing the data will depend upon the
objectives of the individual monitoring program
- differences in lipid analysis techniques will result in differences
in lipid detected - caution in comparing data between analytical
methods
• Time of Sampling
- hydrophobic contaminants are mobilized and transported to the
egg during oogenesis
- timing of sampling should be based on biological cycles which
influence an organism's susceptibility to bioaccumulate
- it is recommended that target species be sampled when tissue
contamination concentrations are expected to be at their highest
levels
Existing Analytical Methods
• It is recommended that consistent types of analytical protocols be imple-
mented to allow for comparisons among studies
• Metals/Metalloids
- GFA AS is the recommended method for the detection of metals
and metalloids (U.S. EPA, 1987a)
- cold vapor AAS is the recommended protocol for mercury
detection
Organics
- GC/MS in conjunction with isotope dilution is recommended for
the detection of semi-volatile organic compounds
- vacuum super-cooled trap in conjunction with a purge and trap
device is recommended for the detection of volatile organics
(Hiatt, 1981)
-------
B-220 ]
- isotope dilution option is recommended as it provides reliable
recovery data for each analyte (U.S. EPA, 1986a)
QAIQC Considerations
• Reports delineating the essential elements of the bioaccumulation compo-
nent of the program should be included with the quantitative QA/QC
analyses
Statistical Design Considerations
• Six individuals comprising each of five replicate composites should be
adequate to detect a treatment difference equal to 100 percent of the overall
mean among treatments
Use of Data
• Establish spatial and temporal trends in the bioaccumulation of toxicants of
selected estuarine fish and macroinvertebrates
• Identify existing and potential problem areas for fish and macroinverte-
brate contamination
• Supply data which can be used to calculate the human health risk of
consuming estuarine fish and shellfish
ASTM. 1991. Standard guide for collection, storage, characterization, and manipula- B10.8
lion of sediments for toxicological testing. ASTM Designation E1391-90. In: Annual Literature Cited
Book of ASTM Standards. American Society for Testing and Standards, Philadelphia, and References
PA.
Boehm, P.D. 1984. The Status and Trends Program: Recommendations for design
and implementation of the chemical measurement segment. Workshop Report.
Rockvillc,MD:NOAA.
dcBocr, J. 1988. Chlorobiphenyls in bound and non-bound lipids of fishes: Compari-
son of different extraction methods. Chemosphere 17:1803-1810.
-------
B-221
WWW.V.W.'W.
DeWitt,T. 1991. Hatfield Marine Science Center, Newport, Oregon. Personal
communication.
DiToro, D.M, J.D. Mahony, D.J. Hansen, K.J. Scott, A.R. Carlson, and G.T. Ankley.
In Press. Acid volatile sulfide predicts the acute toxicity of cadmium and nickel in
sediments.
Farrington, J.W., E.D. Goldberg, R.W. Risebrough, J.H. Martin, and V.T. Bowen.
1983. U.S. "Mussel Watch" 1976-1978: An overview of the trace-metal, DDE, PCB,
hydrocarbon, and artificial radionuclide data. Environ. Sci. Technol. 17:490-496.
Ferraro, S.P., H. Lee, R.J. Ozretich, and D.T. Specht. 1990. Predicting bioaccumula-
tion potential: A test of a fugacity-based model. Arch. Environ. Contain. Toxicol.
19:386-394.
Fowler, S.W. 1982. Biological transfer and transport processes. In: Pollutant Trans-
fer and Transport in the Sea, vol 2. (Kullenberg, G., ed). Boca Raton, FL: CRC Press
Gardner, W.S., T.F. Nalepa, W.A. Frez, E.A. Cichocki, and P.P. Landrum. 1985.
Seasonal patterns in lipid content of Lake Michigan macroinvertebrates. Can. J. Fish.
Aquat. Sci. 42:1827-1832.
Gardner, W.S., W.A. Frez, E.A. Cichocki, and C.C. Parrish. 1986. Micromethod for
lipids in aquatic invertebrates. Limnol. Oceanogr. 30:1099-1105.
Goldberg, E.D., V.T. Bowen, G.H. Farrington, J.H. Martin, P.L. Parker, R.W.
Risebrough, W. Robertson, E. Schneider and E. Gamble. 1978. The mussel watch.
Environ. Conserv. 5:101-125.
Hansen, P.D., H. Von Westerhagen, and H. Rosenthal. 1985. Chlorinated hydrocar-
bons and hatching success in Baltic herring spring spawners. Mar. Environ. Res.
15:59-76.
Hiatt, M.H. 1981. Analysis of fish and sediment for volatile priority pollutants. Anal.
Chem. 53:1541-1543.
Karickhoff, S.W., D.S. Brown, and T.A. Scott. 1979. Sorption of hydrophobic
pollutants on natural sediments. Wat. Res. 13:241-248.
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B-222 ]
lB»w
Knczovich, J.P. and F.L. Harrison. 1987. A new method for determining the concen-
tration of volatile organic compounds in sediment interstitial water. Bull. Environ.
Contam. Toxicol. 38:837-940.
Ladd, J.M., S.P. Hayes, M. Martin, M.D. Stephenson, S.L. Coale, J. Linfield, and M.
Brown. 1984. California state mussel watch: 1981-1983. Trace metals and synthetic
organic compounds in mussels from California's coast, bays, and estuaries. Biennial
Report. Sacramento, CA: Water Quality Monitoring Report No. 83-6TS.
Lake, J.L., N.I. Rubinstein, and S. Pavignano. 1987. Predicting bioaccumulation:
Development of a partitioning model for use as a screen tool in regulating ocean
disposal of wastes. In: Fate and Effects of Sediment-bound Chemicals In Aquatic
Systems. (Dickson, K.L., A.W. Maki, and W.A. Brungs, eds). Florissant, CO: Sixth
Pellston Workshop.
Landrum, P.P. and J.A. Robbins. 1990. Bioavailability of sediment-associated
contaminants to benthic invertebrates. In: Sediments: Chemistry and Toxicitv of In-
Place Pollutants (Baudo, R., J.P. Giesy, and H. Muntau, eds). Lewis Publishers
Lee, H., II, B.L. Boese, J. Pelletier, M. Winsor, D.T. Specht, and R.C. Randall. 1989.
Guidance manual: Bedded sediment bioaccumulation tests. EPA ERLN Document
Nlll, Pacific Ecosystems Branch, Newport, OR.
Lee, H., II, M. Winsor, J. Pelletier, R.C. Randall, J. Bertling and B. Coleman. 1990.
Computerized risk and bioaccumulation system (Ver. 1.0). EPA ERLN Document
N137, Pacific Ecosystems Branch, Newport, OR.
Niimi, A.J. 1983. Biological and toxicological effects of environmental contaminants
in fish and their eggs. Can. J. Fish. Aquat. Sci. 40:306-312.
Pearson, T.H. and R. Rosenberg. 1978. Macrobenthic succession in relation to
Organic enrichment and pollution of the marine environment. Oceanogr. Mar. Biol.
Ann. Rev. 16:229-311.
Phillips, DJ.H. 1980. Quantitative Aquatic Biological Indicators. London, England:
Applied Science Publ. Ltd.
Phillips, DJ.H. and D.S. Segar. 1986. Use of bio-indicators in monitoring conserva-
tive contaminants: Programme design imperatives. Mar. Poll. Bull. 17:10-17.
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Plumb, R.H. 1981. Procedure for handling and chemical analysis of sediment and
water samples. Technical Report EPA/CE-81-1. U.S. EPA and Corps of Engineers,
U.S. Army Engineers Waterways Experimental Station, Vicksburg, MS.
Rubinstein, N.I., J.L. Lake, R.J. Pruell, H. Lee, B. Taplin, J. Heltshe, R. Bowen and S.
Pavignano. 1987. Predicting bioaccumulation of sediment-associated organic con-
taminants: Development of a regulatory tool for dredged material evaluation. Internal
Report. Narragansett, RI: US EPA.
Spies, R.B., D.W. Rice and J. Felton. 1988. Effects of organic contaminants on
reproduction of the starry flounder Platichthys stellatus in San Francisco Bay. Mar.
Biol. 98:181-189.
U.S. EPA. 1982. Method for use of caged mussels to monitor for bioaccumulation
and selected biological responses of toxic substances in municipal wastewater dis-
charges to marine waters. Draft. Environmental Monitoring Science Laboratory,
Cincinnati, OH.
U.S. EPA. 1985a. Bioaccumulation monitoring guidance: Selection of target species
and review of available bioaccumulation data. vol. 2. EPA 403/9-86-006. Office of
Marine and Estuarine Protection, Washington, D.C. 52 pp.
U.S. EPA. 1985b. Bioaccumulation Monitoring Guidance: Recommended Analytical
Detection Limits, vol. 3. EPA 503/6-90-001. Office of Marine and Estuarine Protec-
tion, Washington, D.C. 23 pp.
U.S. EPA. 1985c. Bioaccumulation monitoring guidance:Estimating the potential for
bioaccumulation of priority pollutants and 301(h) pesticides discharged into marine
and estuarine waters, vol 1. EPA 503/3-90-001. Office of Marine and Estuarine
Protection, Washington, D.C. 56 pp.
U.S. EPA. 1986a. Bioaccumulation Monitoring Guidance: Analytical methods for
U.S. EPA priority pollutants and 301(h) pesticides in tissues from estuarine and marine
organisms. Vol.4. EPA 503/6-90-002. Office of Marine and Estuarine Protection,
Washington, D.C. 108pp.
U.S. EPA. 1986b. Test methods for evaluating solid wastes, physical/chemical
methods. SW-846, 3rd Edition. Environmental Protection Agency, Washington, D.C.
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U.S. EPA. 1987a. Quality Assurance/Quality Control (QA/QC) for 301(h) Monitor-
ing Programs: Guidance on Field and Laboratory Methods. EPA 430/9-86-004.
Office of Marine and Estuarine Protection, Washington, D.C.
U.S. EPA. 1987b. Puget Sound protocols. Final Report. 31pp. Prepared for
Region X, Office of Puget Sound.
U.S. EPA. 1987c. Bioaccumulation Monitoring Guidance: Strategies for Sample
Replication and Compositing, vol.5. EPA 430/9-87-003. Office of Marine and
Estuarine Protection, Washington, D.C. 51 pp.
U.S. EPA. 1987d. Technical support document for ODES statistical power analysis.
EPA 430/9-87-005. Office of Marine and Estuarine Protection, Washington, D.C. 34pp.
U.S. EPA. 1989. Assessing human health risks from chemically contaminated fish
and shellfish: A guidance manual. EPA 503/8-89-002. Office of Marine and Estuarine
Protection, Washington, D.C. 136 pp.
U.S. EPA. 1990a. Assessment and Control of Bioconcentratable Contaminants in
Surface Waters, (draft). Office of Water Enforcement and Permits, Washington, D.C.
U.S. EPA. 19905. Analytical procedures and quality assurance plan for the determi-
nation of xenobiotic chemical contaminants in fish. EPA 600/3-90-023. Office of
Research and Development, Environmental Research Laboratory, Duluth, MN. 23 pp.
U.S. EPA. 1990c. Statement of work for organics analysis: multi-media, multi-
concentration. Document No. OLM01.0. U.S. Environmental Protection Agency,
Contract Laboratory Program. Washington, D.C.
U.S. EPA. 1991. Statement of work for inorganic analysis: multi-media, multi-
concentration. Document No. DLM02.0. U.S. Environmental Protection Agency,
Contract Laboratory Program. Washington, D.C.
Young, D.R., AJ. Mearns, and R.W. Gosset. 1990. Bioaccumulation and
biomagnification of DDT and PCB residues in a benthic and a pelagic food web of
Southern California.
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B11.0 Bacterial and Viral Pathogens
Monitoring for pathogenic microorganisms is currently conducted by state environ-
mental and human health agencies in shellfish harvesting areas and bathing beaches.
National Pollution Discharge Elimination System (NPDES) pathogen monitoring
programs may be underway, at selected locations in the estuary, in order to assess both
the condition of water in the vicinity of discharges and surrounding areas and to assess
relative pathogen contributions from these permitted effluent discharges.
The National Shellfish Sanitation Program (NSSP) has been established as a joint
effort between the Food and Drug Administration (FDA), state agencies, and the
shellfish industry to set forth guidelines for the management of state shellfish pro-
grams. As part of the NSSP, FDA provides technical assistance to states in studying
specific pollution problems; provides data to establish closure levels for shellfish
harvesting; conducts applied research on various contaminants to assist in developing
standards and criteria; and evaluates the effectiveness of state shellfish sanitary control
programs. In addition, since 1966, data have been compiled periodically by FDA and
the National Oceanic and Atmospheric Administration (NOAA) on classification by
states of coastal and estuarine waters with regard to suitability for shellfishing activi-
ties. In addition to classifying their waters as to their suitability as shellfish harvesting
areas, states also issue beach closures. These closures are typically based on water
quality criteria developed by the Federal government.
B11.1 Human pathogens found in the estuarine environment include viruses and bacteria.
Rationale In the United States, viruses and bacteria are the most important human pathogens,
both in terms of the number of organisms released to the environment and in the
severity of the diseases they cause (OTA, 1987).
Humans can be exposed to pathogens by direct contact with contaminated waters (e.g.,
swimming, surfing, diving) and/or indirectly through ingestion of contaminated food
(e.g., molluscan shellfish). The preponderance of evidence indicates that the etiologic
agent for waterborne outbreaks of acute gastroenteritis (AGI) are the Norwalk-like
viruses (Kaplan et al., 1982). Hepatitis A has been linked to the consumption of raw
or partially cooked molluscan shellfish (Feingold, 1973; CDC, 1979; Ohara etal,
1983). Bacteria responsible for typhoid and cholera are known to be water- and
seafood-borne. These pathogens can enter the estuarine environment through the
discharge of raw sewage, wastewater effluent from sewage treatment plants, failing
septic tanks, and the dumping of sewage sludge (OTA, 1987).
Monitoring of human pathogens provides information essential in relating the presence
of pathogens in marine waters and shellfish to outbreaks of disease. Monitoring data
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B-2261
1 * „ *, S V\
may be used to identify potential sources of pathogens. The assessment of pathogen
contamination should be a component of a monitoring program where pathogens may
present a risk to human health and economic vitality of an estuary.
Water Column Sampling
Bacteria are not uniformly distributed throughout the water column (Gameson, 1983);
bacterial abundances, several orders of magnitude greater than underlying waters, can
be found in a thin microlayer on the surface of the water (Hardy, 1982). If feasible, it
is recommended that samples of this microlayer be collected; separate samples of
underlying waters should also be taken. However, standardized methods for sampling
this microlayer have not been established. If it is not feasible to sample both the
microlayer and underlying waters, the "scoop" method should be used to ensure that
the surface microlayer is sampled (U.S. EPA, 1978).
Water samples for bacterial analyses are frequently collected using sterilized plastic
bags (e.g., Whirl-pak) or screw-cap wide-mouthed bottles. Several depths may be
sampled during one cast and/or replicate samples may be collected at a particular depth
using a Kemmerer or Niskin sampler (U.S. EPA, 1978). Any device which collects
water samples in unsterilized tubes should not be used for collecting bacteriological
samples without first obtaining data that support its use. Pumps may be used to
sample large volumes of the water column (U.S. EPA, 1978).
Sentinel Organisms
Analyses of sentinel shellfish tissue (e.g., mussels and oysters) offer several advan-
tages:
• they concentrate pathogens; they may be useful for viral analyses since
viruses are often present in low numbers
• they provide a means of temporally integrating water quality conditions
• they may be direct measures of human exposure to pathogens
(i.e., consumption of contaminated food)
• they may be deployed and maintained in a number of diverse locales
B11.2
Monitoring Design
Considerations
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B-227
,
^:"
However, the disadvantages of sentinel organisms include:
• the cost of transplanting organisms
• the possibility of loss of the cage/buoy system
• species may not be tolerant of all test site conditions (e.g., low salinity,
high turbidity)
It is essential that monitoring design elements be standardized to allow for compari-
sons among estuarine studies. The selection of a standard sentinel species is recom-
mended; interspecies differences would not allow comparisons of body burdens.
B11.3 Many pathogens can be present in wastes, contaminated media, and infected organ-
Analytical isms. Human pathogens are of concern in estuarine waters because they are:
Methods
Considerations • associated with major debilitating diseases (e.g., hepatitis, cholera)
• infectious at low doses
• resistant to environmental stress
• not readily enumerated due to low numbers
Table Bl 1-1 provides examples of pathogenic organisms known to cause adverse
human health effects. Due to difficulties in culturing and identifying pathogens that
occur in very low abundances, standardized protocols have not been developed for
their identification and enumeration. The alternative to directly identifying pathogens
of concern has been the enumeration of bacteria which are indicators of human waste
contamination or indicators of human illness.
Indicators of Human Health Risks
Due to the inability to enumerate pathogens of concern, indicators of human pathogen
densities have been used in order to assess human health risks. Indicator organisms
useful in predicting infectious disease rates should have the following characteristics:
• high abundances should be consistently found in human fecal wastes
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B-228I
«* "
X
jww wfjwvfjwff
TABLE
MICROORGANISMS RESPONSIBtCFOR CAUSING ADVERSE
HUMAN HEALTH EFFECTS* (NOAA, 1988)
Disease
Pathogenic
Organism
Seafood
Source
Hepatitis Hepatitis A virus
Non-A and non-B hepatitis
•• '*'* x
Gastroenteritis Aeromenas hydrophilia \ s
and Plesiomonas^shigettaides"
Vibrio
Vibrio parahaentiolyticus
Vibrio •
Vibrio cholera 0 group -T
•. H "X
Vibrio cholera, Non-O group I
Norwalkvirus ,\ „,, ,"
Raw oysters
Steamed and raw clams^.
, Cockles;
*••>•• s s
Raw molltiscan shellfish
Shellfish
Raw oysters
Clams and sna||s
Raw oysters
Crab
Shrimp
Lobster
Raw oysters ;
Kaw'oysters
boiled shrimp, bailed crab
"Raw oysters - - -J
Raw oysters
Small round structured vims „ ^Raw oysters
Campylobacterjejunt^f'^ \^-T3RawClaras" ••'"^
* Includes naturally occurring microorganisms a$ well as microorganisms associated
with pollution.
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B-229
* s-i^f" ' '^WPP^"? '
Xfc-Xv. •.
• should not have significant extra-human fecal sources
• must provide temporally and spatially reliable and accurate appraisals of
the pathogen of concern
Currently, there is no consensus on which indicator organism, if any, is specific for
human feces.
Coliform Bacteria - For decades, the concentration of coliform bacteria, either total or
fecal coliform, has been considered a reliable indicator of the presence and densities of
pathogens. The use of total coliform bacterial criteria to protect human health from
disease associated with contaminated water is widespread. Furthermore, total coliform
concentrations have the advantage of providing a basis for comparison with historical
data (U.S. EPA, 1988).
However, these indicators and their corresponding water quality standards have not
been related to incidences of disease through epidemiological studies. Controversy
exists on the efficacy of using coliform bacteria to predict the presence of other types
of pathogens (Pederson, 1980). In fact, recent studies indicate that fecal coliform
bacteria may not be a reliable indicator for predicting the risks associated with direct
exposures to pathogens in the marine environment (Cabelli et al., 1979 and 1982).
Fecal coliforms are not pathogenic, and are less resistant to environmental stress
compared to many pathogens (Borrego et al., 1983). Furthermore, fecal coliform
bacteria are not specific to mammalian fecal pollution. The lack of this specificity
prompted the development of methods for enumerating fecal coliform bacteria specific
to mammalian fecal pollution (e.g., Escherichia coli). However, with respect to
recreational water quality criteria, the EPA has not recommended the use of E. coli for
marine and estuarine waters.
Enterococci - Enterococci are streptococcus bacteria indigenous to the intestines of
warm-blooded animals. Cabelli et al. (1983) found that the densities of enterococci
were highly correlated with the incidence of gastrointestinal symptoms (GI) among
swimmers; reported swimming-associated GI symptoms were poorly correlated with
fecal coliform densities. Enterococci also have the following advantageous character-
istics:
tolerant to high salinity and are of particular value in the analysis of
estuarine and marine waters
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B-230!
• taxonomic identifications are relatively simple and can reveal the kinds of
mammalian pollution (e.g., humans, livestock)
• genetic fingerprinting techniques have been developed which can link
these bacteria in the environment to specific sources of contamination
State-of-the-art genetic fingerprinting techniques are very costly and require further
study in order to assess their reliability. The EPA has adopted enterococci as an
indicator of microbiological water quality for recreational marine waters.
Viruses - Viruses are being recognized as major etiologic agents for many outbreaks
of human illness as a result of swimming in contaminated water and consumption of
contaminated seafood. Viruses are excreted only by infected individuals; they are not
normal flora in the intestinal tract. Traditional indicators of microbial contaminants
appear to be inadequate for predicting human health risks associated with both con-
suming molluscan shellfish and swimming in waters that contain sewage associated
viruses. However, examination of water for enteric viruses is not recommended at this
time, except in special circumstances, due to limitations of methodologies. Even state-
of-the-art methods for concentrating viruses from water still are being researched and
continue to be modified and improved. None of the available virus detection methods
have been tested adequately with representatives from all of the virus groups of public
health importance. In addition, some of these methods require expensive equipment
and materials for sample processing and all virus assay and identification procedures
require expensive cell culture and related virology laboratory facilities.
Laboratory Techniques
It should also be noted that no single procedure is adequate to isolate all microorgan-
isms from water, and the presence of one microorganism does not signify the presence
or absence of any other (Table B11-2). A more detailed description of analytical
methods for a number of pathogens is presented in Standards Methods for the Exami-
nation of Water and Wastewater CAPHA. 1989).
Fecal Bacteria - Two standard methods are presented here for the detection of fecal
bacteria: membrane filter procedure, and multiple-tube fermentation procedure.
The membrane filter (MF) technique involves sample filtration followed by direct
plating for detection and enumeration of coliform bacterial densities. The MF tech-
nique can be used to test relatively large volumes of samples, and yields numerical
results more rapidly than the multiple-tube procedure. The statistical reliability of the
-------
B-231
TABLE BM-2.
LABORATORY PROCEDURES FOR BACTERIAL INDICATOR^
Laboratory Procedures
Test Organisms!
Sediment
Tissue
Fecal colifbm
bacteria - -
Fecalcolifonn -
MPN tubes using A-I
broth (APHA, 1989)-
(fecal coliform
MPN tubes using A-I
broth CAPHA* 1989)
(fecal colifonn
bacteria/100 ml)
MPN tubes using EC
broth (APHA, 19B9)
(fecal colifoiin
bacteria/100 ml) -
Enterococci
mTEC (DuFour <# ,
1981)(E.cbnyiQOml)
mB (Levin «-«t» 1975)a
(enterococci/100 ml)
rciCP by membrane
isson and
ml)
mCP by membrane
filtration (Bmeison and
Cabelli, 1982)
(C
MPN tubes using iron
milk(St John y membrane filtrafion foT%water (Bisson and
Cabelli 1975) andsedlment (Emerson and Cafcelli 1982), and teon roilk tube using MPN techniqii (St John- ^
)» The Bisson and Cabelli (1979) method is preferred^ s "^
MF technique is greater than the Most Probable Number (MPN) procedures (APHA,
1989). However, the MF technique has limitations, particularly in testing waters with
high turbidity and noncoliform (background) bacteria. The MF technique can be used
to measure bacterial densities of Escherichia coli (E. coli) and enterococci in ambient
waters (U.S. EPA, 1985); the EPA has approved this technique for use in seawater.
The multiple-tube fermentation technique examines a series of fermentation tubes
containing growth media which are inoculated with the appropriate decimal dilutions
of water (multiples and submultiples of 10 ml), based on the probable coliform den-
sity. Formation of gas in any amount of time constitutes a preliminary positive for the
presence of coliform bacteria; confirmation phase analyses are then conducted. The
precision of the test depends on the number of tubes examined in each dilution.
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B-2321
Adequate mixing of the sample in order to yield a homogenous dispersion of bacteria
in the test portions is critical to the accuracy of the test; clumping of bacteria will
result in underestimations of actual bacterial density. This technique is commonly
used in assessing bacterial levels in shellfish.
The most probably number (MPN) index is an index of the number of coliform bacte-
ria that, most probably, would produce the results observed in the laboratory examina-
tion (APHA, 1989). Values for the MPN index can be obtained from standard tables
based on the results of the multiple-tube fermentation technique or from Thomas'
formula (APHA, 1989).
Viruses - Detecting viruses in estuarine waters requires collecting a representative
water sample, concentrating viruses in the sample, and identifying and estimating
abundances of these concentrated viruses. Difficulties in detecting virus abundances
include:
• viruses are very small (20-100 nm)
* virus concentrations in waters are spatially and temporally variable, and
typically low
• dissolved and suspended material in the sample interferes with accurate
virus detection
Because virus concentrations may be very low, significant volumes of water may be
required (e.g., on the order of tens to thousands of liters). Dose-response curves are
lacking for most pathogens; however, as few as 10 to 100 bacteria are capable of
inducing disease (OTA, 1987).
Three different techniques for concentrating and enumerating viruses are presented in
Standard Methods (APHA. 1989):
• adsorption to and elution from microporous filters
• aluminum hydroxide adsorption and precipitation
• polyethylene glycol hydroextraction-dialysis
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B-233
The adsorption and elution technique pressure-filters viruses on microporous filters
and then elutes them from the filter in a small liquid volume. Generally, two types of
filters are available: electronegative and electropositive filters. Currently, insufficient
documentation on the efficacy of electropositive filters exists. Limitations to this
technique include:
• clogging of adsorbent filter by suspended material
• dissolved colloid material may interfere by competing with viruses for
adsorption sites on the filter
• viruses adsorbed to suspended material may be removed during suspended
material clean-up procedures
Methods for recovering solids-associated viruses are found in Standard Methods
(APHA, 1989). In spite of these limitations, the adsorption and elution technique
remains the most promising technique for detecting viruses.
The aluminum hydroxide adsorption and precipitation, and the polyethylene glycol
hydroextraction-dialysis techniques are used to reconcentrate viruses in proteinaceous
and organic buffer eluates. These methods may be used to concentrate viruses in
waters having high virus densities (e.g., wastewaters). They may also be used as a
second-step concentration procedure following processing of large fluid volumes
through microporous filters. However, these two techniques are impractical for
primary processing of large fluid volumes (APHA, 1989).
New Techniques - Traditional techniques lack the ability to detect apparently viable,
but non-culturable microorganisms. Recently developed monoclonal antibody and
gene probe techniques permit the detection and enumeration of both culturable and
non-culturable microorganisms. Since these methods have the ability to detect non-
culturable organisms, they may serve as more precise techniques for monitoring
microbiological water quality. Furthermore, these new techniques will allow direct
monitoring of pathogens of concern; the promise of monitoring specific pathogens,
such as Salmonella, Shigella, Giardia, or Legionella may soon be realized.
Sample Handling
Preservation and storage of water samples can introduce significant sources of error.
Sample bottles must be resistant to sterilization procedures.
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B-2341
Samplcs should be refrigerated (1-4°C) during transport to a laboratory and analyzed
within six hours of collection (U.S. EPA, 1978).
It is recommended that sterile distilled water be transported to the field, transferred to a
sample bottle, and processed routinely to ensure that samples were not contaminated
during collection and transport. It is also recommended that ten percent of the samples
be analyzed in duplicate. Furthermore, ten percent of the samples should be split and
analyzed by two or more laboratories.
Intra-laboratory and inter-laboratory quality control practices should be documented
and QA/QC reports should be available for inspection. Further recommended quality
assurance guidelines for a microbiology laboratory are available (Inhom, 1977;
Bordner et a/., 1978) and are also discussed in Standard Methods (APHA, 1989).
B11.4
QA/QC
Considerations
Bacterial counts often are characterized as having a skewed distribution because of
many low values and a few high ones (APHA, 1989). Application of parametric
statistical techniques requires the assumption of symmetrical distributions such as the
normal curve. An approximately normal distribution can be obtained from positively
skewed data by converting numbers to their logarithms (APHA, 1989). Accordingly,
the preferred statistic for measuring central tendency of microbiological data is the
geometric mean.
Consideration of statistical strategies will mitigate the high costs of collecting and
processing samples. Analyses of power-costs are necessary in selecting appropriate
sample/replicate number, sample location and sampling frequency (Ferraro et al,
1989). See also Statistical Design Considerations: Composite Sampling, Statistical
Power, and Power-Cost Analysis (Section B.3).
B11.5
Statistical Design
Considerations
The assessment of pathogen contamination should be a component of a monitoring
program where pathogens may be present a risk to human health and the economic
vitality of an estuary. Monitoring of human pathogens provides information essential
in relating the temporal and spatial distribution of infectious agents in estuarine waters
and shellfish to the epidemiology of pathogens of concern (e.g., affected human
populations, locations, and timing of the outbreak).
B11.6
Data Use
-------
B-235
Furthennore, monitoring data can be used to identify discharges which may be signifi-
cant sources of pathogens and to ensure that water quality standards are maintained.
Monitoring data can also be used to verify fate and transport, human health risk
assessment, and epidemiological modeling predictions.
B11.7
Summary and
Recommendations
Rationale
• The objective is to detect and describe spatial and temporal changes in
abundances of indicators of human health pathogens
• Monitoring indicators of human pathogens provides information essential
in relating the presence of infectious agents in estiiarine waters and shell-
fish to the incidence of disease outbreaks and potential sources of these
agents.
Monitoring Design Considerations
• If feasible, samples of the surface microlayer should be collected separate
from samples of underlying waters. If not feasible, samples containing
both underlying and microlayer waters should be collected. It is highly
recommended that consistent types of sampling protocols be implemented
to allow for comparisons among studies
• Analysis of tissues of sentinel organisms (e.g., mussels and oysters)
confers the following advantages:
- they concentrate pathogens
1 - they provide a means of temporally integrating water quality
conditions
- they may be deployed and maintained in a number of locales
Analytical Methods Considerations
• Indicators of human pathogens should have the following characteristics:
- be consistently found in high abundance in human fecal wastes
- should not have significant extra-human fecal sources
- must provide temporally and spatially reliable and accurate
appraisals of the pathogen of concern
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B-2361
• Fecal Coliform Bacteria Densities
- may not be a reliable indicator for predicting the risk associated
with direct exposures to pathogens
- this measure provides a means to compare to historical data
- Laboratory analyses include membrane filter technique and
multiple-tube fermentation technique
• Enterococci Densities
- are highly correlated with the incidence of gastrointestinal
symptoms
- taxonomic identifications are relatively simple and can reveal the
kinds of mammalian pollution
- state-of-the-art genetic fingerprinting techniques can link bacteria
in the estuary to specific sources; however these techniques are
very costly and require further study in order to assess their
reliability
- Laboratory analyses include membrane filter technique
• Viral Densities
- recognized as major etiologic agents for many outbreaks of
human illness
- examination of water samples for enteric viruses is not recom-
mended due to limitations of methodologies
- Laboratory analyses include absorption to and elution from
microporous filters, aluminum hydroxide absorption-precipita-
tion, and polyethylene glycol hydroextraction-dialysis
• Monoclonal antibodies and gene probes show promise in detecting and
enumerating both culturable and non-culturable microorganisms
QAIQC Considerations
• Sterile distilled water should be transported and transferred to sample
bottles in order to assess contamination during collection and transport
• Ten percent of the samples should be analyzed in duplicate
• Ten percent should be split and analyzed by two or more laboratories
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B-237
Statistical Design Considerations
• Preferred statistic for measuring central tendency of microbiological data is
the geometric mean
Use of Data
• Provide essential information in order to assess threats to human health
• Establishes temporal and spatial trends in pathogen densities and identifies
potential relationships between the presence of the indicator and incidence
of human illness
• Provides data which can be used to verify fate and transport, human health
risk, and epidemiology models
B11.8 Andrews, W.H. and M. W. Presnell. 1972. Rapid recovery of Escherichia coli from
Literature Cited estuarine water. Appl. Microbiol. 23:521.
and References
APHA. 1989. American Public Health Association, American Water Works Associa-
tion, and Water Pollution Control Federation, Standard methods for the examination of
water and wastewater, 17th Edition. American Public Health Association, Washing-
ton, D.C.
Bisson, J.W. and V.J. Cabelli. 1979. Membrane filtration enumeration method for
Clostridiwnperfringens. Appl. Environ. Microbiol. 37:55-66.
Bitton, G., B.N. Feldberg, and S.R. Farrah. 1979. Concentration of enteroviruses from
seawater and tap water by organic flocculation using non-fat dry milk and casein
water. Air Soil Pollut. 10:187.
Booze-Allen and Hamilton. 1983. A background document on pathogenic organisms
commonly found in municipal sludge. Prepared for ECAO, US EPA. Cincinnati, OH.
Bordner, R.H., J.A. Winter, and P.V. Scarpino, eds. 1978. Microbiological Methods for
Monitoring the Environment, Water and Waste. EPA 600/8-78-017. Environmental
Monitoring and Support Lab., U.S. Environmental Protection Agency, Cincinnati, Ohio.
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B-238 |
Borrego, J.J., F. Arrabal, A. de Vicente, L.F. Gomez, and P. Romero. 1983. Study of
microbial inactivation in the marine environment. J. Water Pollut. Control Fed.
55:297-302.
Brezenski, F.T. and J.A. Winter. 1979. Use of the delayed incubation membrane filter
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