&EPA
 U.S. Army Corps
 of Engineers
             United States
             Environmental Protection
             Agency
              Department of The Arniy
              US Army Corps of Engineers
EPA-823-B-94-002
June 1994
             Office of Water (4305)
Evaluation of Dredged Material
Proposed For Discharge in
Waters of the U.S. - Testing
Manual (Draft)

Inland Testing Manual

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               EVALUATION OF DREDGED MATERIAL
PROPOSED FOR DISCHARGE IN WATERS OF THE U.S. - TESTING MANUAL
                            (DRAFT)

                   (INLAND TESTING MANUAL)
                           Prepared by

              ENVIRONMENTAL PROTECTION AGENCY
                         Office of Water
                   Office of Science and Technology
                        Washington, D.C.

                             and

                   DEPARTMENT OF THE ARMY
                United States Army Corps of Engineers
                        Washington, D.C.

                           June  1994

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                                        11
                           TABLE OF CONTENTS
                                                                          Page No.

Title Page                                                                         i
Table of Contents                                                                   ii
List of Tables                                                                     vi
List of Figures                                                                   vii

PREFACE                                                                        1
DEFINITIONS                                                                    4
LIST OF ACRONYMS                                                            12
CONVERSIONS                                                                  16

PART I • GENERAL CONSIDERATIONS  	   17

1.0         INTRODUCTION	   18
1.1         Background  	   18
1.2         Statutory/Regulatory Overview	   18
1.2.1        Statutory Overview  	   18
1.2.2        Section 404 Regulatory Overview  	   19
1.2.2.1                   The Section 404(b)(l) Guidelines  	  21
1.2.2.2                   Particulars of Sections 230.50 and 230.61   	  22
1.2.3        Relationship to Section 401 CWA Water Quality Certification 	  23

2.0         SCOPE AND APPLICABILITY	  25
2.1         This Manual is Intended to Address:	  25
2.2         This Manual is Not Intended to Address:	  25
2.3         Dredged Material Discharge for Beneficial Uses	  26
2.4         The Role of Biological Evaluations (Toxicity and/or Bioaccumulation Tests)
            in the Manual	  27
2.5         The Role of Water and Sediment Chemical Evaluations in the Manual	  28
2.6         Water Column Effects  	  29
2.7         Mixing	  29
2.8         Benthic Effects  	  29
2.9         Management Options	  30
2.10        The Relationship of the  Inland Testing Manual to Other USACE/EPA
            Dredged Material Management Efforts  	  30
2.10.1       Relationship of the Manual to the USACE/EPA Framework Document  ....  30
2.10.2       Relationship of the Manual to the EPA/USACE Green Book  	  31
2.10.3       Relationship of the Manual to EPA's Contaminated Sediment Strategy and
            Sediment Quality Criteria	  31

PART II - EVALUATION OF POTENTIAL ENVIRONMENTAL IMPACT	  33
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                                          Ill

3,0          OVERVIEW OF TESTING AND EVALUATION	   34
3.1          Tiered Testing and Evaluation	   34
3.2          Control and Reference Sediments 	   35
3.2.1         Reference Sediment Sampling	   39
3.2.2         Reference Sediment Sampling Plan	   40

4.0          TIER I EVALUATION	   42
4.1          Compilation of Existing Information	   44
4.2          Identification of Contaminants of Concern	   47
4.2.1         Microbial Contamination	   48
4.2.2         Chemical Contamination	   48
4.3          Tier I Conclusions	   52

5.0          TIER II EVALUATION	   55
5.1          Water Column Impact  	   55
5.1.1         Screen Relative To WQS	   56
5.1.2         Elutriate Analysis Relative To WQS  	   56
5.2          Benthic Impact  	   56
5.3          Tier II Conclusions  	   57

6.0          TIER III EVALUATION  	   59
6.1          Water Column Toxicity Tests   	   60
6.2          Benthic Toxicity Tests	   60
6.3          Benthic Bioaccumulation	   61
6.4          Tier III  Conclusions	   65

7.0          TIER IV EVALUATION  	   69
7.1          Toxicity Tests	   69
7.2          Benthic Bioaccumulation	   70

PART III - SAMPLING AND ANALYSIS	   71

8.0          SAMPLING	   72
8.1          Preparation For Sampling	   72
8.2          Components Of A Sampling Plan	   75
8.2.1         Review of Dredging Plan	   75
8.2.2         Historical Data	   76
8.2.3         Subdivision of Dredging Area	   77
8.2.4         Selection of Sampling Locations and Number of Samples	   78
8.2.5         Sample Collection Methods 	   83
8.2.5.1                    Sediment Sample Collection   	   83
8.2.5.2                    Water Sample Collection  	   85
8.2.5.3                    Organism Collection  	   85
8.2.6         Sample Handling, Preservation, and Storage	   85
8.2.6.1                    Sample Handling	   85
8.2.6.2                    Sample Preservation  	   91
8.2.6.3                    Sample Storage	   92
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                                          IV

8.2.7         Logistical Considerations and Safety Precautions	   92
8.2.8         Non-Indigenous Test Species  	   93

9.0          PHYSICAL ANALYSIS OF SEDIMENT AND CHEMICAL ANALYSIS OF
             SEDIMENT, WATER, AND TISSUE SAMPLES  	   95
9.1          Physical Analysis of Sediment   	   95
9.2          Target Detection Limits  	   96
9.3          Chemical Analysis of Sediment	   97
9.3.1         Target Analytes	   97
9.3.2         Selection of Analytical Techniques	  100
9.4          Chemical Analysis of Water	  107
9.4.1         Analytical Targets	  107
9.4.2         Analytical Techniques	  107
9.5          Chemical Analysis of Tissues	  109
9.5.1         Target Analytes	  109
9.5.2         Analytical Techniques	  113

10.0         GUIDANCE FOR PERFORMING TIER II EVALUATIONS  	  117
10.1         Tier II:  Water Column Effects  	  117
10.1.1        Screen Relative To WQS	  117
10.1.2        Elutriate Analysis Relative To WQS  	  118
10.1.2.1                   Standard Elutriate Preparation  	  118
10.1.2.2                   Chemical Analysis	  119
10.1.2.3                   Comparison with WQS (Standard Elutriate Test)  	  119
10.2         Theoretical   Bioaccumulation  Potential  (TBP)  of  Nonpolar  Organic
             Chemicals	  120

11.0         GUIDANCE FOR PERFORMING BIOLOGICAL  EFFECTS TESTS	  127
11.1         Tier III:  Water Column Toxicity Tests	  127
11.1.1        Species Selection  	  128
11.1.2        Apparatus  	  130
11.1.3        Laboratory  Conditions  	  130
11.1.4        Laboratory  Procedures	  131
11.1.5        Data Presentation and Analysis	  132
11.1.6        Conclusions  	  133
11.2         Tier III: Benthic Toxicity Tests  	  134
11.2.1        Species Selection  	  134
11.2.2        Laboratory  Procedures	  137
11.2.3        Chronic/Sublethal Tests  	  140
11.2.4        Data Presentation and Analysis	  141
11.2.5        Conclusions  	  141
11.3         Tier IV: Chronic/Sublethal Effects Evaluations	  142
11.4         Tier IV: Case Specific Evaluations  	  142

12.0         GUIDANCE FOR PERFORMING BIOACCUMULATION TESTS  	  145
12.1         Tier III:  Determination Of Bioavailability	  145
12.1.1        Species Selection and Apparatus	  145
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12.1.2
12.1.3
12.1.4
12.1.5
12.2
12.2.1
12.2.2
12.2.2.1
12.2.2.2
12.2.2.3
12.2.2.4
12.2.2.5
12.2.2.6
12.2.2.7
12.2.2.8

13.0
Experimental Conditions	  147
Chemical Analysis	  148
Data Presentation and Analysis	  148
Conclusions  	  149
Tier IV:  Determination Of Steady State Bioaccumulation	  149
Laboratory Testing  	  149
Field Assessment of Steady State Bioaccumulation	  ISO
            Apparatus	  151
            Species Selection	  151
            Sampling Design and Conduct	  152
            Basis for Evaluation of Bioaccumulation	  152
            Sample Collection and Handling  	  152
            Chemical Analysis	  153
            Data Presentation and Analysis 	  153
            Conclusions	  153
REFERENCES	  154
APPENDICES
APPENDIX A:
APPENDIX B:

APPENDIX C:
APPENDIX D:
APPENDIX E:

APPENDIX F:

APPENDIX G:
       40 CFR PART 230
       GUIDANCE FOR EVALUATION OF EFFLUENT DISCHARGES FROM
       CONFINED DISPOSAL FACILITIES
       EVALUATION OF MIXING
       STATISTICAL METHODS
       SUMMARY OF TEST CONDITIONS  AND  TEST ACCEPTABILITY
       CRITERIA FOR TIER III BIOASSAYS
       METHODOLOGIES FOR IDENTIFYING AMMONIA AS A TOXICANT
       IN DREDGED-MATERIAL  TOXICITY TESTS
       QUALITY   ASSURANCE/QUALITY   CONTROL  (QA/QC)
       CONSIDERATIONS
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                                            VI
                                  LIST OF TABLES
Table 1.
Industries Associated with Sediment Contaminants.
Page No.

      46
Table 2.       Food  and Drug Administration  (FDA)  Action  Levels for Poisonous and
              Deleterious Substances in Fish and Shellfish for Human Food.                    63

Table 3.       Type of Samples Which May Be Required Following Tier I to Conduct Dredged-
              Material Evaluation Tests.                                                   73

Table 4.       Summary of Recommended Procedures for Sample Collection, Preservation, and
              Storage.                                                                    86

Table 5.       Potential Contaminants of Concern Listed According to  Structural Compound
              Class.                                                                     98

Table 6.       PCDD and PCDF Compounds Determined by Method 1613                     102

Table 7.       Polychlorinated Biphenyl (PCB) Congeners Recommended  for Quantitation  as
              Potential Contaminants of Concern.                                          103

Table 8.       Methodology for Toxicity Equivalency Factors                                 106

Table 9.       Octanol/Water Partition  Coefficients  (K^)  for Organic Compound  Priority
              Pollutants and 301 (h) Pesticides.                                             110

Table 10.      Bioconcentration Factors (BCF) of Inorganic Priority Pollutants.                 112

Table 11.      Candidate Toxicity Test Species for Determining Potential Water Column Impact
              of Dredged Material Disposal.                                               129

Table 12.      Candidate Acute Toxicity Test Species for Determining Potential Benthic Impact
              of Dredged-Material Disposal.                                               135

Table 13.      Candidate Test Species for Determining Potential Bioaccumulation from Whole
              Sediment Tests.                                                            146
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                                            Vll
                                 LIST OF FIGURES
                                                                                 Page No.
Figure 1.      Simplified  Overview of Tiered Approach to Evaluating  Potential Impact of
              Aquatic Disposal of Dredged Material.                                         36

Figure 2.      Illustration of Tiered Approach to Evaluating Potential Water Column Impacts of
              Dredged Material.                                                           37

Figure 3.      Illustration of Tiered Approach to Evaluating  Potential  Benthic Impacts of
              Deposited Dredged Material.                                                 38

Figure 4.      Nomograph for Determining Theoretical Bioaccumulation Potential.              123
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Vlll
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                                         PREFACE
The "Evaluation of Dredged Material Proposed for Discharge in Waters of the U.S. - Testing Manual".
commonly referred to as the Inland Testing Manual represents a major effort by the U.S. Army Corps
of Engineers  (USAGE)  and the Environmental Protection Agency (EPA) to establish procedures
applicable to the evaluation of potential contaminant-related environmental impacts associated with the
discharge of dredged material in inland waters, near coastal waters, and surrounding environs (that is,
all waters other than the ocean and the territorial seas, regulated pursuant to Section 404, CWA). This
manual is consistent, to the maximum extent practicable, with the procedures established for ocean waters
(i.e., the "Green Book" entitled "Evaluation of Dredged Material Proposed for Ocean Disposal - Testing
Manual" - EPA/USACE, 1991). The USAGE and EPA have statutory and regulatory responsibilities with
regard to the management of dredged material discharge activities in inland and near coastal waters.  The
USAGE  is responsible for regulating non-Federal dredging and dredged material  discharge activities
through a permit program, and for conducting Federal dredging and dredged material discharge activities
in conjunction with its Civil Works Program. EPA is responsible for establishing,  in conjunction with
the USAGE, guidelines pertaining to the evaluation of these activities, and performing oversight actions.
Specifically, Section 404 of the Federal Water Pollution Control Act of 1972 (FWPCA), Public Law 92-
500, as  amended by the Clean Water Act of 1977 (CWA), Public Law 95-217, requires, among other
things, that the discharge of dredged or fill material into waters of the U.S. be permitted by the USAGE.
The USAGE also conducts Civil Works dredging and dredged material discharge activities in accordance
with Section 404. Section 404 further requires that discharge sites be specified though the application of
the Section 404(b)(l) Guidelines (Guidelines) developed by EPA in conjunction with the USAGE. Section
404 requires that the "guidelines shall be based upon criteria comparable to the criteria applicable to the
territorial seas, contiguous zone, and the ocean". Thus,  a clear connection for comparable testing for
ocean, inland and near coastal waters was established as early as 1972.

The Guidelines, which impart other requirements in addition to those associated with contaminant-related
impacts, are published at 40 CFR 230. This manual provides testing procedures applicable to determining
the  potential for contaminant-related environmental impacts associated with the discharge of dredged
material. Dredged material evaluated under the procedures described in this manual must also  satisfy all
other applicable requirements of 40 CFR 230-232, 33  CFR 320-330, and 33 CFR 335-338 in order to
comply  with the Guidelines and to be authorized for discharge.

This manual,  which is designed  to allow for regional  flexibility in implementation and application
including development of regional manuals and documentation, will be periodically revised and updated
as warranted by advances in regulatory practice and technical understanding. This manual replaces the
May 1976 proposed testing protocol, "Ecological Evaluation of Proposed Discharge of Dredged or Fill
Material Into Navigable Waters", which will no longer be applicable. The 1976 protocol was developed
in response to  a requirement in the Federal Register notice of the Guidelines, Vol. 40, No. 173, Friday,
5 September 1975. That notice states the "EPA in conjunction with the Corps of Engineers will publish
a procedures manual that will cover summary and description of tests, definitions, sample collection and
preservation, procedures, calculations and references."  In December 1980, the Guidelines were revised
and finalized in the Federal  Register Vol. 45, No. 249.  The present joint effort by EPA and USAGE
contains up-to-date testing procedures to implement the Guidelines at Sections 230.60 and 230.61, and
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is intended to bring compatibility and a comparable level of environmental protection for dredged material
testing in ocean, inland and near coastal waters.

This manual is one of a series of guidance documents jointly developed by EPA and the USAGE
pertaining to  dredged  material disposal.  This  series includes  a document  entitled  "Evaluating
Environmental Effects of Dredged Material Management Alternatives  - A Technical Framework"
(Framework Document - USACE/EPA, 1992). The Framework Document articulates those factors to
be considered in identifying the environmental effects of dredged material management alternatives on
a continuum of discharge sites from uplands to the oceans (management alternatives include open water,
confined  and beneficial use situations) that  meet the substantive and  procedural  requirements of the
National  Environmental Policy Act (NEPA), the  CWA  and the  Marine Protection,  Research, and
Sanctuaries Act (MPRSA). The Green Book (EPA/USACE, 1991) is included in the series. Application
of the testing guidance in this manual in addition to guidance provided in the Framework Document and
the Green Book will allow for consistency in decision making with  respect to technical considerations,
across statutory boundaries and the continuum of dredged material discharge options.

The contributions made by many individuals from both agencies are gratefully acknowledged. The first
and second drafts of the manual were completed by the Environmental  Laboratory (EL) of the USAGE
Waterways Experiment Station (WES):  Thomas Wright, primary author; Michael Palermo, author of
Appendix B; Paul Schroeder, Michael Palermo, Robert Randall and  Billy Johnson, authors of Appendix
C. Succeeding drafts were completed by an EPA/USAGE Workgroup established by EPA's Office of
Science and Technology (OST) within the Office of Water (OW). Mike Kravitz of OST was the Work
Assignment Manager. Appendix D was written by Dennis Brandon and  Joan Clarke (WES) and Michael
Paine (EVS Consultants). Appendix F was written by Gary Ankley  (EPA). Appendix G was written by
Sandra Salazar and  Peter Chapman (EVS Consultants). Henry Lee and Bruce Boese (EPA) contributed
valuable information pertaining to  sediment bioaccumulation testing.

The Workgroup was comprised of individuals from  headquarters, field offices and research laboratories
of both agencies with scientific and/or programmatic experience related to dredged material discharge
activities.

Co-Chairs:     Betsy Southerland            EPA/OW/OST
              Kirk Stark                   USACE/Headquarters
Members:     Gary Ankley                EPA/Duluth Research Lab
              Tom Dillon                 USACEAVES
              Wade Eakle                 USACE/San Francisco Dist.
              Robert Engler               USACE/WES
              John Goodin                EPA/OW/OWOW
              Mike Kravitz                EPA/OW/OST
              John Malek                 EPA/Region 10
              David Mathis                USACE/Headquarters
              Jan Miller                   USACE/North Central Div.
              Michael Palermo             USACE/WES
              William Peltier              EPA/Region 4
              David Redford              EPA/OW/OWOW
              Susan Ivester Rees           USACE/Mobile District
              James Reese                 USACE/North Pacific Div.
              Brian Ross                  EPA/Region 9
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              Norm Rubinstein             EPA/Narragansett Lab
              Dave Tomey                 EPA/Region 1
              Joe Wilson                   USACE/Headquarters
              Thomas Wright               USACE/WES
              Howard Zar                  EPA/Region 5

Contractor:    Peter Chapman               EVS Consultants

Review of this manual was conducted by EPA through OW [OST and the Office of Wetlands, Oceans
and Watersheds (OWOW)] and by USAGE through the Office of the Chief of Engineers (Regulatory
Branch, Dredging and Navigation Branch, Office of Environmental Policy) and EL of WES. In addition,
the results of the EPA's Science Advisory Board (SAB, 1992) review of the 1991 Green Book were
considered in detail, where applicable, during development of this manual. The results of EPA's SAB
(1994) review of the present draft Inland Testing Manual were considered during its finalization. Regional
issues which have National relevance were provided by EPA Region and USAGE Division and District
staff,  and were incorporated into  the appropriate sections of this  document.  This manual provides
comprehensive testing guidance from a national perspective. Within the framework of this document,
EPA  Regions and USAGE  Districts and  Divisions will  develop  region-specific guidance and/or
procedures, as necessary (e.g., region-specific test species), to provide sufficient information to make
informed dredged material discharge decisions.
This manual should be cited as follows:

EPA/USACE.  1994. Evaluation of dredged material proposed for discharge in waters of the U.S. -
Testing manual (Draft). EPA-823-B-94-002, Washington, D.C.
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                                      DEFINITIONS
The following definitions of words and terms are specific to the use of this manual and, where applicable,
are quoted verbatim from the Guidelines  (cf.  Definitions  at 40 CFR 230.3 and/or other parts; such
definitions are starred*). The importance of definitions has long been recognized by the American Society
for Testing and Materials (e.g., Pratt, 1991) whose Committee on Terminology frequently invoke the
Humpty Dumpty quotation, "When I use a word, it means just what I choose it to mean - neither more
nor less" (Schindler, 1991). Thorough familiarization with the following definitions is required prior to
use of this manual.

Accuracy: The ability to obtain a true value; determined by the degree of agreement between an observed
       value and an accepted reference value.

Acid volatile sulfide (AYS): The sulfides removed from sediment by cold acid extraction, consisting
       mainly of H2S and FeS. AVS is a possible predictive tool for divalent metal sediment toxicity.

Acute: Having a sudden onset, lasting a short time.

Acute toxicity:   Short-term toxicity to organism(s) that  have been affected by  the  properties  of a
       substance,  such as  contaminated  sediment.  The  acute toxicity  of a sediment is  generally
       determined by quantifying the mortality of appropriately sensitive organisms that are put into
       contact with the sediment, under either field or laboratory conditions, for a specified period.

* Adjacent: Bordering, contiguous or neighboring. Wetlands separated from other waters of the United
       States by man-made dikes or barriers, natural river berms, beach dunes and the like are "adjacent
       wetlands".

Application factor (AF): A numerical, unitless value, calculated as the threshold chronically toxic
       concentration of a test substance divided by its acutely toxic concentration. The AF is  usually
       reported as a range and is multiplied by the median lethal concentration as determined in a short-
       term (acute) toxicity test to estimate an expected no-effect concentration under chronic exposure.

Benchmark organism: Test organism designated by USAGE and EPA as appropriately sensitive and
       useful for determining biological data applicable to the real  world.  Test protocols with  such
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       organisms are published, reproducible and standardized; they can be performed by any competent
       bioassay laboratory irrespective of geographic location.

Bioaccumulation:  The accumulation of contaminants in the tissue of organisms  through any route,
       including respiration, ingestion, or direct contact with contaminated water, sediment, pore water
       or dredged material. [The regulations require that bioaccumulation be considered as part of the
       environmental evaluation of dredged material proposed for disposal. This consideration involves
       predicting whether there will be a cause-and-effect relationship between an organism's presence
       in the area influenced by the dredged material and an environmentally important elevation of its
       tissue content or body burden of contaminants above that in similar animals not influenced by the
       disposal of the dredged  material].

Bioaccumulation factor: The degree to which an organism accumulates a chemical compared to the
       source. It is a dimensionless  number  or  factor derived  by dividing the concentration in the
       organism by that in the  source.

Bioassay: A bioassay is  a test  using  a biological system. It involves exposing an organism to a test
       material and determining a response. There are two major types of bioassays differentiated by
       response: toxicity tests which measure  an effect (e.g., acute toxicity, sublethal/chronic toxicity)
       and bioaccumulation tests which measure a phenomenon (e.g.,  the uptake of contaminants into
       tissues).

Bioavailable:  Can be taken up by organisms,  i.e., from water, sediment, suspended particles, food.

Bioconcentration:  Uptake of a substance  from water.

Biomagnification:  Bioaccumulation up the food chain, e.g., the route of accumulation is solely through
       food.  Organisms at higher trophic levels will  have higher body burdens than those at lower
       trophic levels.

Biota sediment accumulation factor:  Relative concentration of a substance in the tissues of an organism
       compared to the concentration of the same substance in the sediment.

Bulk sediment chemistry:  Results of chemical analyses of whole sediments (in terms of wet or dry
       weight), without normalization (e.g., to organic carbon, grain-size, acid volatile sulfide).

Can: Is used to mean "is able to".
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Chronic: Involving a stimulus that is lingering or which continues for a long time.

Chronic toxicity:  See sublethal/chronic toxicity.

Comparability: The confidence with which one data set can be compared to others and the expression
       of results consistent with other organizations reporting similar data. Comparability of procedures
       also implies using methodologies that produce results comparable in terms of precision and bias.

Completeness: A measure of the amount of valid data obtained versus the amount of data originally
       intended to be collected.

Confined  disposal: A disposal method that isolates the dredged material from the environment. Confined
       disposal is placement of dredged material within diked confined disposal facilities via pipeline or
       other means.

Confined  disposal  facility (CDF):  A diked  area, either in-water or upland, used to contain dredged
       material. The terms confined disposal facility (CDF), dredged material containment area, diked
       disposal facility, and confined disposal area are used interchangeably.

Constituents:  Chemical substances, solids, liquids, organic matter, and organisms  associated with or
       contained in or on dredged material.

•Contaminant: A  chemical or biological substance in a form that can be incorporated into, onto or be
       ingested by and that harms aquatic organisms, consumers of aquatic organisms, or users of the
       aquatic environment, and includes but is not limited to the substances on the 307(a)(l) list of toxic
       pollutants promulgated  on January 31, 1978 (43 FR 4109). [Note:  A contaminant  that causes
       actual harm is technically referred to as a pollutant, but the regulatory definition of a "pollutant"
       in the Guidelines is different, reflecting the intent of the CWA.]

Contaminant of concern:  A contaminant present in a given sediment thought to have the potential for
       unacceptable adverse environmental impact due to a proposed discharge.

Control sediment:  A sediment essentially free of contaminants and compatible with the biological needs
       of the test organisms such that it has no discernable influence on the  response being measured in
       the test. Control sediment may be the sediment from which the test organisms are collected or
       a laboratory sediment, provided the organisms meet control standards. Test procedures are
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       conducted with the control sediment in the same way as the reference sediment and dredged
       material. The purpose of the control sediment is to confirm the biological acceptability of the test
       conditions and to help verify the health of the organisms during the test. Excessive mortality in
       the control sediment indicates a problem with the test conditions or organisms, and can invalidate
       the results of the corresponding dredged material test.

Data quality indicators:  Quantitative statistics and qualitative descriptors which are used to  interpret
       the degree of acceptability or utility of data to the user; include bias (systematic error), precision,
       accuracy, comparability, completeness, representativeness, detectability and statistical confidence.

Data quality objectives (DQOs):  Qualitative and quantitative statements of the overall uncertainty that
       a decision maker is willing to accept in results or decisions derived  from environmental data.
       DQOs provide the framework for planning environmental data operations consistent with the data
       user's needs.

Discharge of dredged material:  Any addition of dredged material into waters of the United States.
       [Dredged  material discharges  include:  open water  discharges;  discharges  resulting  from
       unconfmed disposal operations (such as beach nourishment or other beneficial uses); discharges
       from confined disposal facilities which enter waters of the United States (such as effluent, surface
       runoff, or leachate);  and,  overflow  from dredge hoppers, scows, or other transport  vessels].
       Material resuspended during normal  dredging operations is considered "de minimus" and is not
       regulated under Section 404 as a dredged material discharge.  See 33 CFR 323.2 for a detailed
       definition. The potential impact of resuspension due to dredging can be addressed under NEPA.

'Disposal site:  That portion of the "waters of the United States" where  specific disposal activities are
       permitted and consist of a bottom surface area and any overlying volume of water. In the case
       of wetlands on which surface water is not present, the disposal site consists of the wetland surface
       area. [Note: upland locations, although not mentioned  in this definition in the Regulations, can
       also be disposal sites].

District: A USAGE administrative area.

*Dredged material: Material that  is excavated or dredged from waters of the United States.  [A general
       discussion of the nature of dredged material is provided by Engler et  al. (1991a)].
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	8	

ECM: The median effective concentration. The concentration of a substance that causes a specified effect
       (generally sublethal rather than acutely lethal) in 50% of the organisms tested in a laboratory
       toxicity test of specified duration.

Elutriate: Material prepared from the sediment dilution water and used for chemical analyses and toxicity
       testing. Different types of elutriates are prepared for two  different procedures as noted in this
       manual.

Evaluation:  The process of judging data in order to reach a decision.

*Factual determination: A determination in writing of the potential short-term or long-term effects of
       a proposed discharge of dredged or fill material  on the physical,  chemical and biological
       components of the aquatic environment in light of Subparts C-F of the Guidelines.

Federal Standard: The dredged material disposal  alternative(s) identified by the U.S. Army Corps of
       Engineers that represent the least costly, environmentally acceptable alternative(s) consistent with
       sound engineering practices and which meet the environmental standards  established  by the
       404(b)(l) evaluation process. [See Engler et al. (1988) and 33 CFR 335-338].

*Fill material: Any material used for the primary purpose of replacing an aquatic area with dry  land or
       changing the bottom elevation of a water body for any purpose.   The term does not include any
       pollutant discharged into the water  primarily to  dispose of waste, as  that activity is regulated
       under Section 402 of the Clean Water Act. [Note: dredged material can be used as fill material].

Grain-size effects:  Mortality or other effects  in laboratory toxicity tests  due to sediment granulometry,
       not chemical toxicity. It is clearly best to use test organisms which are not likely to react to grain-
       size but, if this is not reasonably possible, then testing must account for any grain-size effects.

Guidelines:  Substantive environmental criteria by  which proposed discharges of dredged material are
       evaluated. CWA Section 404(b)(l) final rule (40 CFR 230) promulgated December 24,  1980.

LCjo: The median lethal concentration.  The concentration of a substance that kills 50% of the organisms
       tested in a laboratory toxicity test of specified duration.

Leachate:  Water or any other liquid that  may contain  dissolved (leached) soluble materials,  such as
       organic salts and mineral salts, derived from a solid material.
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Lethal: Causing death.

Loading density:  The ratio of organism biomass or numbers to  the volume of test solution in an
       exposure chamber.

Management  actions:  Those actions considered necessary to rapidly render harmless the material
       proposed for discharge (e.g., non-toxic, non-bioaccumulative) and which may include containment
       in or out of the waters of the U.S. (see 40 CFR Subpart H). Management actions are employed
       to reduce adverse impacts of proposed discharges of dredged material.

Management unit: A manageable, dredgeable unit of sediment which can be differentiated by sampling
       and which  can be separately dredged and disposed within a larger dredging  area. Management
       units are not differentiated solely on physical or other measures or tests but are also based on site-
       and project-specific considerations.

May:  Is used to mean "is allowed to".

Method detection  limit (MDL): The minimum concentration of a  substance which can be identified,
       measured, and reported with 99% confidence that the analyte concentration is greater than zero.

Might: Is used to mean "could possibly."

*Mixing zone: A limited volume of water serving as a zone of initial dilution in the immediate vicinity
       of a discharge point where receiving water quality may not meet quality  standards or other
       requirements otherwise applicable to the receiving water.  [The mixing zone  may be defined by
       the volume and/or the surface  area of the disposal site or specific mixing zone definitions in State
       water quality standards].

Must: In this manual refers to requirements that have to be addressed in the context of compliance with
       the Guidelines.

Open water disposal: Placement of dredged material in rivers, lakes or estuaries via pipeline or surface
       release from hopper dredges or barges.

Pathway: In the case of bioavailable contaminants, the route of exposure (e.g., water, food).
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'Pollution: The man-made or man-induced alteration of the chemical, physical, biological or radiological
       integrity of an aquatic ecosystem. [See definition of contaminant].

'Practicable: Available and  capable of being  done  after  taking into consideration  cost, existing
       technology, and logistics in light of overall  project purposes.

Practical quantitation  limit  (PQL):  The lowest concentration that can be reliably quantified with
       specified limits of precision and accuracy during routine laboratory operating conditions.

Precision: The ability to replicate a value; the degree to which observations or measurements of the same
       property, usually obtained under similar conditions, conform to themselves. Usually expressed
       as standard deviation,  variance or range.

QA: Quality assurance, the total integrated program for assuring the reliability of data. A system for
       integrating the quality planning,  quality control, quality assessment, and quality improvement
       efforts to  meet user  requirements  and defined standards  of quality with  a  stated level  of
       confidence.

QC: Quality control, the overall  system of technical activities for obtaining prescribed standards of
       performance in the monitoring and measurement process to meet user requirements.
                       i»
Reason to believe: Subpart G of the 404(b) (1) guidelines requires the use of available information to
       make a preliminary determination concerning the need for testing of the material proposed for
       dredging. This  principle is  commonly known as "reason  to believe",  and  is used in Tier I
       evaluations to determine acceptability of the material for discharge without testing. The decision
       to not perform additional testing based on prior information must be documented, in order to
       provide  a "reasonable assurance that the  proposed discharge material  is  not a  carrier  of
       contaminants" (230.60(b)).

Reference sediment: A sediment, substantially free of contaminants, that is as similar as practicable to
       the grain size of the dredged material and the sediment at the disposal site, and that reflects the
       conditions that would  exist in the vicinity of the disposal site had no dredged-material disposal
       ever taken place, but had all other influences on sediment condition taken place. These conditions
       should be met to the maximum extent possible. For waters of the United States it is recognized
       that background levels of contaminants from sources other than dredged material discharges may
       be substantial and  that consequently,  in some cases  (e.g.,  when the whole area within which
       dredging  and discharge occur is contaminated), additional clarification on this issue  may  be
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       provided in Regional applications. The reference sediment serves as a point of comparison to
       identify potential effects of contaminants in the dredged material. [NOTE: The reference sediment
       concept is the subject of a CWA Section 404 ndemaldng under development.  The rule will be
       proposed in the Federal Register in 1994 for public comment and will be promulgated prior to
       issuance of the final Inland Testing Manual.]

Reference site: The location from which reference sediment is obtained.

Region:  An EPA administrative area.

region: A geographical area.

Regulations:  Procedures and concepts published in the Code of Federal Regulations for evaluating the
       discharge of dredged material into waters of the United States.

Representativeness: The degree to which  sample  data depict an existing environmental  condition; a
       measure of the total variability associated with sampling and measuring that includes the two
       major error components:  systematic error (bias) and random error. Sampling representativeness
       is  accomplished through proper selection of sampling locations and  sampling techniques,
       collection of sufficient number of samples, and use of appropriate subsampling  and handling
       techniques.

Sediment: Material, such as sand, silt, or clay, suspended in or settled on the bottom of a water body.
       The term dredged material refers to material which has been dredged from a water body, while
       the term sediment refers to material in a water body prior to the dredging process.

Should:  Is used to state that the specified condition is recommended and ought  to be met unless there
       are clear and definite reasons not to do so.

Standard operating procedure (SOP): A written document which details an operation, analysis, or
       action  whose mechanisms are thoroughly prescribed and which  is commonly  accepted as the
       method for performing certain routine or repetitive tasks.

Standardized:  In the case of methodology, a published procedure which has been peer reviewed (e.g.,
       journal, technical report), and generally accepted by the relevant technical  community of experts.
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Sublethal: Not directly causing death; producing less obvious effects on behavior, biochemical and/or
       physiological function, histology of organisms.

Sublethal/chronic toxicity:  Biological tests which use such factors as abnormal development, growth
       and reproduction, rather than solely lethality, as end-points. These tests involve all or at least an
       important, sensitive portion of an organism's life-history. A sublethal endpoint may result either
       from short-term or long-term (chronic) exposures.

Target detection limit:  A performance goal set by consensus between the lowest, technically feasible,
       detection limit for routine analytical methods and available regulatory criteria or guidelines for
       evaluating dredged material. The target detection limit is, therefore, equal to or greater than the
       lowest amount of a  chemical that can be reliably detected based on the variability of the blank
       response of routine analytical methods.  However, the reliability  of a chemical  measurement
       generally increases as the concentration increases.  Analytical costs may also be lower  at higher
       detection limits.   For these reasons, a target detection limit is typically set at  not less than  10
       times lower than available dredged material guidelines.

Tests/testing: Specific procedures which generate biological, chemical,  and/or physical data to be used
       in  evaluations.  The data  are usually quantitative but may  be qualitative (e.g.,  taste, odor,
       organism behavior). Testing for discharges of dredged material in waters of the United States is
       specified at 40 CFR  230.60 and 230.61 and is implemented through the procedures  in this
       manual.

Tiered approach: A structured, hierarchical procedure for determining  data needs relative to  decision-
       making, which involves a series of tiers or levels of intensity of investigation. Typically, tiered
       testing involves decreased uncertainty and increased available information with increasing tiers.
       This approach is intended to ensure the maintenance and protection of environmental quality, as
       well as the optimal use of resources. Specifically, least effort is required in situations where clear
       determinations can be made of whether (or not) unacceptable adverse impacts are likely to occur
       based on available information.  Most effort is required where  clear determinations cannot be
       made with available information.

Toxicity:  see Acute toxicity; Sublethal/chronic toxicity, Toxicity test.

Toxicity test: A bioassay which measures an effect (e.g., acute toxicity,  sublethal/chronic toxicity). Not
       a bioaccumulation test (see definition of bioassay).
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Water quality certification: A state certification, pursuant to Section 401 of the Clean Water Act, that
       the proposed discharge of dredged material will comply with the applicable provisions of Sections
       301, 303, 306 and 307 of the Clean Water Act and relevant State laws. Typically this certification
       is provided by the affected  State.  In instances where the State lacks jurisdiction (e.g.,  Tribal
       Lands), such  certification is provided  by EPA or the Tribe (with an approved certification
       program).

Water quality standard: A law or regulation that consists of the beneficial designated use or uses of a
       water body, the numeric and narrative water quality criteria that are necessary to protect the use
       or uses of that particular water body, and an anti-degradation statement.

Waters of the U.S.:  In general,  all waters landward of the baseline of the territorial sea and the
       territorial sea. Specifically, all waters defined  in Section  230.3  (s)  of  the  Guidelines. [See
       Appendix A].

Whole sediment: The sediment and interstitial waters  of the proposed dredged  material or reference
       sediment before it has undergone any processing that might alter its chemical or lexicological
       properties. For purposes of this manual, press-sieving to remove organisms from test sediments,
       homogenization of test sediments,  compositing of sediment samples, and  additions of small
       amounts  of water to facilitate homogenizing or compositing sediments may be necessary to
       conducting bioassay tests. These procedures are considered unlikely to substantially alter chemical
       or lexicological properties of the respective whole sediments  except in the  case of AVS (acid
       volatile sulfide) measurements (EPA, 199la) which are not presently required. Alternatively, wet
       sieving, elutriation, or freezing and thawing of sediments may alter chemical and/or lexicological
       properties, and sediment so processed should nol be considered as  whole sediment for bioassay
       purposes.
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                                           14
                              LIST OF ACRONYMS
AAS - Atomic Absorption Spectroscopy
AF - Application Factor
AVS - Acid Volatile Sulfide
BAF - Bioaccumulation Factor
BCF - Bioconcentration Factor
BSAF - Biota Sediment Accumulation Factor
CDF - Confined Disposal Facility
CFR - Code of Federal Regulations
CLP - Contract Laboratory Program
CWA - Clean Water Act
BCD - Electron Capture Detection
EO - Executive Orders
EPA - Environmental Protection Agency
FDA - Food and Drug Administration
FR - Federal Register
GC - Gas Chromatography
GFAAS - Graphite Furnace Atomic Absorption Spectroscopy
IAEA - International Atomic Energy Agency
ICP - Inductively Coupled Plasma
ITM - Inland Testing Manual
LBP - Lipid Bioaccumulation Potential
MPRSA - Marine Protection, Research and Sanctuaries Act
MS - Mass Spectrometry
NBS - National Bureau of Standards
NEPA - National Environmental Policy Act
NIST - National Institute for Standards and Technology
NOAA - National Oceanic Atmospheric Administration
NPDES - National  Pollutant Discharge Elimination System
NRC - National Research Council of Canada
PAH - Polynuclear Aromatic Hydrocarbons
PCB - Polychlorinated Biphenyl
QA - Quality Assurance
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QC - Quality Control
QSAR - Quantitative Structure Activity Relationship
RHA - Rivers and Harbors Act of 1899
SAB - Science Advisory Board
SIM - Selected Ion Monitoring
SOP - Standard Operating Procedure
SQC - Sediment Quality Criteria
SQS - Sediment Quality Standards
SRM - Standard Reference Material
TBP - Theoretical Bioaccumulation Potential
TDL - Target Detection Limit
TOC - Total Organic Carbon
TIE - Toxicity Identification Evaluation
USAGE - U.S. Army Corps of Engineers
USCS -  Unified Soil Classification System
WQC - Water Quality Criteria
WQS - Water Quality Standards
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                                        16
                                CONVERSIONS
      METRIC TO IMPERIAL
                                                  IMPERIAL TO METRIC
WEIGHT:
      IKg =  lOOOg = 2.2051b
      Ig = lOOOmg = 2.205 x 10%
      1 mg = lOOOjig = 2.205 x
                                                  lib = 16 oz = 0.4536Kg
LENGTH:
      1m = 100cm = 3.28 ft. = 39.370in
      1cm = 10mm = 0.3937in
      1mm = 1000/ig = 0.03937in
                                                  1 foot (ft) = 12in = 0.3048m
CONCENTRATION:
      Ippm = Img/L = Img/Kg = l/*g/g = ImL/m3
      Ig/cc = IKg/L = 8.3454 Ib/gallon (US)
      lg/m3 = Img/L = 6.243 x 10-5lb/ft3

VOLUME:
      1L = lOOOmL
      ImL = 1000/tL
      Ice = lO-'m3
                                                  1 Ib/gal = 7.4811b/ft3 = 0.120g/cc =
                                                     119.826g/L = 119.826Kg/m3
                                                  1 oz/gal = 7.489Kg/m3
                                                  lyd3 = 27ft3 = 764.555 L = 0.7646m3
                                                  1 acre-ft  = 1233.482m3
                                                  1 gallon (US) = 3785cc
                                                  1 ft3 = 0.0283m3= 28.3168 L
FLOW:
      Im/s = 196.850 ft/min = 3.281 ft/s
      1 m3/s = 35.7 ft3/s
AREA:
      1 m2 = 10.764ft2
      1 hectare (ha) = 10000m2 = 2.471 acres
                                                  1 ftVs = 1699.011 L/min = 28.317 L/s
                                                  1 ft2/hr = 2.778 x 10"4 ft2/s = 2.581 x
                                                     10-5m2/s
                                                  1 ft/s = 0.03048m/s
                                                  1 yd3/min = 0.45ft3/s
                                                  yd'/s = 3.366 gal/s = 12.743 L/s
                                                  1 ft2 = 0.0929m2
                                                  1 acre = 4046.856m2 = 0.405 ha
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              17
PART I - GENERAL CONSIDERATIONS
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1.0           INTRODUCTION

1.1           Background

The "Evaluation of Dredged Material Proposed for Discharge in Waters of the U.S. - Testing Manual".
commonly referred to as the Inland Testing Manual, updates and replaces  "Ecological Evaluation of
Proposed Discharge of Dredged or Fill Material into Navigable Waters"  (USAGE, 1976). This updated
manual contains technical  guidance for determining the potential for contaminant-related impacts
associated with the discharge of dredged material in waters regulated under Section 404 of the CWA
(inland waters, near coastal waters, and surrounding environs) through chemical, physical, and biological
evaluations. The technical guidance in the manual is intended for use  by Army Corps of Engineers
(USAGE) and Environmental Protection Agency (EPA) personnel, state regulatory personnel, as well as
dredging permit  applicants  and  others (e.g., scientists, managers, and other involved or concerned
individuals). The results obtained will be utilized within the context of regulatory requirements (discussed
in the following sections), to facilitate  decision-making  with  regard to the management of dredged
material.

Key changes to the 1976 testing protocol include a tiered testing approach, accommodation for sediment
quality standards (SQS), 28-d bioaccumulation testing, comparison of benthic test results with those of
the reference sediment, improved statistics,  improved model  applications, and new test  organisms.
Because this manual is national in scope, the guidance provided is generic and may need to be modified
in certain  instances. Application  of this guidance in some site- and case-specific situations will require
best professional judgment, appropriately  documented.  Permit applicants and others are strongly
encouraged to consult with their  appropriate Regional and District experts for additional guidance.
1.2           Statutory/Regulatory Overview

The following sections provide a discussion of the statutory and regulatory framework of the Federal
programs within which decisions regarding the management of dredged material discharge activities are
made.
1.2.1         Statutory Overview

The USAGE and EPA share the Federal responsibility for regulating the discharge of dredged material.
The Glean Water Act (CWA) governs discharges of dredged material into  "waters of the United States",
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including all waters landward of the baseline of the territorial sea. The Marine Protection, Research, and
Sanctuaries Act (MPRSA) governs the transportation of dredged material seaward of the baseline (in
ocean waters) for the purpose of disposal. In addition, all activities regulated by these statutes must
comply with the applicable requirements of the National Environmental Policy Act (NEPA), as well as
other Federal laws, regulations and Executive Orders which apply to activities involving the discharge
of dredged material.

The CWA was enacted by Congress to "restore and maintain the chemical, physical,  and biological
integrity of the Nation's waters."  The CWA created three permit programs, under Section 401 (as a
certification), Section 402 and Section 404, to regulate the point-source discharge of pollutants into waters
of the U.S. EPA administers Section 402 which established the National Pollutant Discharge Elimination
System (NPDES) Program to regulate discharges of chemicals, heavy metals, and biological wastes,
primarily in  waste water from  industrial processes,  publicly owned sewage treatment works, and
stormwater discharges.  The Section 402 program may be delegated by EPA to the States to administer.
EPA and USAGE each administer specific aspects of Section 404 which established a permit program and
technical guidelines to regulate discharges of dredged or fill material  (dredged material and fill material
disposal sites must be "specified"). States may assume (and most of them have) the program administered
by EPA under Section  401  and must  grant,  deny, or waive certification for activities permitted  or
conducted by USAGE based on the potential impacts to water quality which may result from a discharge
of dredged or fill material to waters of the U.S.

The USAGE also administers a regulatory  program under Section  10 of the Rivers and Harbors Act of
1899 (RHA) which regulates dredging and other construction activities in navigable waters. The USAGE
also operates  a  Federal Civil Works  navigation program in conjunction  with  the  CWA and  with
requirements  established within Congressional authorization and appropriation statutes, which involves
extensive dredging and dredged material discharge  activities. These USAGE programs are operated in
accordance with NEPA which requires, among other things, the analysis and  documentation of potential
primary and secondary impacts, including those associated with dredging and dredged material discharges.
1.2.2         Section 404 Regulatory Overview

The USAGE has die primary responsibility for the Section 404 regulatory permit program [the USAGE
regulatory program also administers Section 10 RHA, as well as Section 103 of the MPRSA  (for the
transport of dredged material to the ocean for the purpose of disposal)] and is authorized, after notice and
opportunity for public comment, to issue permits specifying sites for the discharge of dredged or fill
material. EPA has the primary role in developing the  environmental guidelines, in conjunction  with
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USAGE [the Section 404(b)(l) Guidelines (Guidelines)], by which permit applications must be evaluated.
EPA is also responsible for commenting on proposed  USAGE permits,  prohibiting discharges  with
unacceptable adverse aquatic environmental impacts, approving and overseeing State assumption of the
program, establishing jurisdiction, and interpreting exemptions. Both USAGE and EPA share enforcement
authority.

The USAGE'S evaluation of a Section 404 permit application involves determining whether the proposed
project complies with the Guidelines (40 CFR 230) and USAGE permit regulations (33 CFR 320-330)
which  require a public interest review of the project. [Public interest factors (listed in 33 CFR 320.4)
considered with respect to dredged material contaminant-related impacts include water quality, water
supply and conservation, safety, and fish and wildlife impacts]. A permit is issued provided the proposed
project complies with the Guidelines and is not contrary to the public interest.  The USAGE issues
individual permits  and general permits. Individual permits are issued on a project-by-preject basis  after
the Guidelines compliance and public interest determinations are made for the specific project at issue.
General permits, on the other  hand, are issued  for classes of activities and/or activities  conducted in
certain classes of waters of the U.S. after the USAGE conducts the Guidelines compliance and public
interest reviews and determines that issuance of the general permit will not result in more  than minimal
adverse impacts to the aquatic environment from either a site-specific or cumulative standpoint.  General
permits require little or no reporting, analysis, or paperwork, compared to individual permits.

There  are three types of general permits issued by the USAGE, nationwide permits,  regional general
permits and programmatic general permits. Nationwide permits are issued by the Chief of Engineers and
apply nationwide. Regional permits are issued by District and Division Engineers and are  applicable on
district or  State-wide basis. Programmatic permits are issued (by the  Chief of Engineers, as well as
District and Division Engineers) to other federal, State or local agencies with the  intention of providing
the appropriate level of environmental protection and avoiding unnecessary duplication of effort with the
agency regulatory  activities at issue.

There  are  currently four nationwide permits that pertain to dredging and the  discharge  of dredged
material. One authorizes the discharge and return water from confined disposal areas (provided the
associated dredging is authorized pursuant to Section 10 of the River and Harbor Act of 1899); two other
nationwide permits authorize the dredging  and discharge,  respectively, of up  to  25 cubic  yards of
material; and a fourth authorizes maintenance dredging of existing marina basins (provided  that the
dredged material is deposited on uplands; return water from a confined disposal  area requires separate
authorization pursuant to  Section 404  of the Clean Water Act). As stated  in  the preamble to the
nationwide permit regulations (FR56, 226, November 22, 1991), the USAGE  depends on its districts'
knowledge of potentially contaminated areas and on the discretionary authority of District and Division
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Engineers to develop special conditions and/or require individual permits where contaminated sediments
are present. General permits are not intended to apply to projects involving the dredging or the discharge
of contaminated materials.

USAGE Civil Works activities are conducted in accordance with the Guidelines and the USAGE operation
and maintenance regulations (33 CFR 335-338). The USAGE specifies sites for the discharge of dredged
material in conjunction with its regulatory and civil works responsibilities. (Permits are not actually issued
in conjunction with USAGE discharge activities).
1.2.2.1                       The Section 404(b)(l) Guidelines

The Guidelines provide the substantive environmental criteria used in evaluating proposed discharges of
dredged or fill material into waters of the United States. Fundamental to these Guidelines is the precept
that dredged or fill material should not be discharged  into the aquatic ecosystem, unless it can  be
demonstrated that such a discharge will not have an unacceptable adverse impact either individually or
in combination with  known and/or probable impacts of other activities affecting the  ecosystems  of
concern.

For proposed discharges of dredged material to comply with the Guidelines, they must  satisfy four
requirements found in Section 230.10 as follows. Section 230.10(a) addresses those impacts associated
with the loss of aquatic site functions and  values of the proposed discharge site, by requiring that the
discharge site represent the least environmentally damaging, practicable alternative. Section 230.10(b)
requires  compliance with established legal  standards (e.g., issuance or waiver of a State water quality
certification).  Section 230.10(c)  requires that discharge of dredged material not result in significant
degradation of the aquatic ecosystem. Section 230.10(d) requires that all practicable means be utilized
to minimize for adverse environmental impacts.

Testing as described in this manual is part of the larger evaluation of a proposed discharge activity to
determine its compliance with the Guidelines. Sections 230.60 and 230.61 of the Guidelines provide the
basis  for certain  factual  determinations  with regard to dredged  material discharge activities. Section
230.60 provides for a general evaluation of the material and establishes a framework to determine, based
on existing information on the proposed dredging and discharge sites, whether the material at issue may
be exempted from further testing. If the conditions at 230.60 cannot be met  or are not applicable, the
testing requirements  of Section 230.61  must be applied.  This manual details the testing procedures
outlined  in 230.60 and 230.61. Conclusions reached utilizing this manual will be used to make factual
determinations of the potential effects of a proposed discharge of dredged or fill material on the physical,
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chemical and biological components of the aquatic environment. Such factual determinations are used to
make findings of compliance or noncompliance with relevant parts of Sections  230.10(b) (including
compliance  with established water quality standards)  and  230.10(c)  (determinations  of potential
contaminant-related impacts to aquatic resources). All specifications of discharge sites must also comply
with Section 230.10 (a) and Section 230.10(d). Site monitoring and/or management activities developed
following the use of this manual  may be said to contribute to satisfying the aforementioned requirements
of Section 230.10(d).

Once compliance with the Guidelines is established, information developed utilizing the manual will also
be factored  into the USAGE public interest determination which  is required by  its regulatory permit
regulations for proposed non-Federal dredged material discharge activities, or its determinations required
by the operation and maintenance regulations pertaining to Federal Civil Works  activities.  In making
determinations with regard to its regulatory and  civil works responsibilities, the USAGE considers a
continuum of discharge options, on a project-specific basis, including alternative sites, mitigation and
specific site management and monitoring conditions.  Determination of whether a material, which would
not otherwise comply with the Guidelines or with other USAGE regulatory and civil works requirements,
could be brought into compliance through appropriate management actions or other discharge methods,
is beyond the scope of this manual.
1.2.2.2                       Particulars of Sections 230.60 and 230.61

Reason to Believe - Subpart G of the 404(b)(l) guidelines requires the use of available information to
make a preliminary determination concerning the need for testing of the material proposed for dredging.
This principle is commonly known as "reason to believe", and is used to determine acceptability of the
material for discharge  without further testing.  The  decision to not perform testing based on prior
information must be documented in order to provide a "reasonable assurance that the proposed discharge
material is not a carrier of contaminants" (by virtue of the fact that it is sufficiently removed from sources
of pollution) [230.60(b)]. The reason to believe that no testing is required is based on the type of material
to be dredged and/or its potential to be contaminated. For example, dredged material is most likely to
be free of contaminants  if the material is composed primarily of sand, gravel, or other inert material and
is found in  areas of high current or wave energy [230.60(a)]. In addition, knowledge of the proposed
dredging site proximity to  other sources of contamination, as well  as that gained from previous testing
or through experience and  knowledge of the area to be dredged, may be utilized to conclude that there
is no reason to believe that  contaminants are present [230.60(b)] and, therefore, no need for testing. This
general evaluation comprises procedures found in Tier I of the manual's tiered-testing framework. Tier
I is a comprehensive analysis of all existing and readily available information on the proposed dredging
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project, including all previously collected physical, chemical, and biological data for both the proposed
dredging and discharge sites. A more complete discussion of technical factors to consider with respect
to Sections 230.60(a) and (b) in Tier I is provided in Section 4.0.

Exclusions From Testing - Sections  230.60(c) and (d) provide for specific circumstances in which the
discharge of dredged material which is suspected to be contaminated may be conducted without further
testing. Section 230.60(c) provides that where the proposed discharge and dredging sites are adjacent and
are comprised of similar materials and subject to the same source(s) of contaminants, disposal may be
conducted without  further testing because the discharge is not likely to result  in degradation of the
discharge site, as  long  as  the  potential  spread of contaminants to less contaminated areas can  be
prevented. Section 230.60(d) provides that  the discharge of contaminated  dredged material may  be
conducted without further testing if constraints, acceptable to USAGE and EPA, are available to reduce
contamination to acceptable levels within the discharge  site, and to prevent contaminants from  being
transported beyond the proposed discharge site boundaries.

Conclusions reached with regard to dredged material discharges without testing, in accordance with
Section 230.60,  must be described in the appropriate factual determination.  Even though material may
be excluded from testing under the  manual the water quality certifying  agency may require testing to
demonstrate compliance with state  laws.  Even in  cases where the discharge site  is  adjacent to the
dredging site,  potential differences in contaminant bioavailability may occur.

Reference Site -  The manual requires comparison of testing results between the proposed dredging site
and a reference site (see previous Definitions section). The USAGE and EPA believe that the use of a
reference  site  provides an accurate information  base for predicting cumulative bioaccumulation and
benthic impacts resulting from the discharge  of dredged material.
1.2.3          Relationship to Section 401 CWA Water Quality Certification

Section 401 of the CWA requires that all Federal permits and licenses, including those for the discharge
of dredged material into waters of the United States, authorized pursuant to Section 404 of the CWA,
must be certified as complying with applicable State water quality standards (WQS). The Guidelines at
40 CFR 230.10(b) state in part that "No discharge of dredged or fill material shall be permitted if it:  (1)
Causes or contributes, after consideration of disposal site dilution and dispersion, to violations of any
applicable State water quality standard." This applies at the edge of a State designated mixing zone.
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The process for adoption of State WQS is prescribed at 40 CFR 131. States must issue, condition, deny,
or waive a Water Quality Certification for activities permitted or conducted by USAGE, certifying that
no adverse water quality impacts will occur based on determinations of compliance with applicable State
WQS which have been adopted in accordance with the above regulation. State water quality standards
consist of designated uses, narrative and numeric criteria designed to support those uses, and anti-
degradation provisions. This testing manual is intended  to provide guidance for the dredged material
testing necessary to determine compliance with such State WQS.

States may, at their discretion, include in their State standards policies generally affecting their application
and implementation, e.g. mixing zones (40 CFR 131.13). A mixing zone is a limited volume of water
serving as a zone of initial dilution in the immediate vicinity of a discharge point where receiving water
may not meet quality standards or other requirements otherwise applicable to the receiving water (40 CFR
230.3). Where mixing  zone provisions are part of the State  standards,  the State  should describe the
procedures for defining mixing zones.

According to EPA (1991b), mixing zone concentrations should not exceed acute water quality standards
and, considering likely pathways of exposure, there should be no significant human health risks. For
dredged material discharges which only occur periodically, water quality  standard compliance in the
mixing zone is generally focused on aquatic life, not on human health, which  is  based on long-term
exposures to contaminants.  (Long-term exposures resulting from accumulations  of dredged material at
the disposal site can be evaluated by such means as bioaccumulation tests). Acute or chronic standards
may be appropriate, depending on the duration of discharge and characteristics of the discharge site.

Many States have statutory or regulatory requirements for use of State-owned lands, including aquatic
(marine and freshwater) bedlands. For discharges of  dredged or fill materials into waters of the U.S.
which are also waters of State or State-owned lands, specific requirements (including testing) for "use"
of State lands may exist which need to be considered. The responsible State land-management agency may
be different from the agency which normally issues the  WQS or coastal zone certification. At a minimum,
coordination with the responsible State agency should occur to avoid conflicts with or impacts to existing
and/or future uses of State lands. In parts of the country, cooperative State-federal dredged material or
sediment  management ventures are in place or are being  pursued to  identify  disposal sites, develop
consistent regional management standards, and to monitor maintenance of those standards [e.g., the Puget
Sound  Dredged Material Disposal  Analysis  (State  of  Washington) and  San  Francisco Long-Term
Management Strategy  (LTMS - State  of California)]. These programs are intended to streamline the
regulatory process associated with dredging and dredged material disposal.
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2.0            SCOPE AND APPLICABILITY

This manual is primarily directed towards evaluation of open water discharges. It utilizes both chemical
and biological analyses as necessary, to provide effects-based conclusions within a tiered framework with
regard to the potential for contaminant-related water column, benthic toxicity and benthic bioaccumulation
impacts. The  tiered-testing procedure detailed in Section  3.1 is comprised of four levels (tiers)  of
increasing  investigative intensity which  generate information to  assist in making contaminant-related
determinations. Tiers I and II use existing or easily acquired information and apply relatively inexpensive
and rapid tests to predict environmental effects. Tiers III and IV contain biological evaluations which are
more intensive and require field sampling, laboratory testing, and rigorous data  analysis.
2.1           This Manual is Intended to Address:

       •      contaminant-related impacts associated with discharges of dredged material in open water
              disposal areas, including wetlands.

       •      contaminant-related impacts to surface water and surrounding environs associated with
              dredged material effluent discharged from confined disposal areas. Guidance on evaluation
              of effluent discharges is provided in Appendix B.  As additional testing guidance becomes
              available on issues such as leachate and runoff it will be added to this manual.
2.2           This Manual is Not Intended to Address:

       •      impacts associated with the dredging activity itself.

       •      impacts associated with the discharge of fill material. Fill material, by its very nature, is
              not usually a significant carrier of contaminants and indeed, a standard USAGE permit
              condition  states  that the  fill  material authorized to be  discharged  shall be  free  of
              contaminants; most concern is with physical effects. An exception to this would be where
              dredged material is used for fill and there is a reason  to believe that contaminants may
              be released. In such an instance, the procedures of this manual are applicable.

       •      microbiological impacts except for impacts in conjunction  with the State designated use
              of a waterbody and human health considerations. The manual provides a list of applicable
              references, as the technology for analyzing other potential impacts from microorganisms
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              (e.g., modeling potential pathways of contamination) is in various stages of development.
              Although scientifically accepted mechanisms  for predicting the degree of potential
              microbiological impacts are not yet available, site management techniques are available
              (but are beyond the scope of this manual) to address potential impacts (e.g., aerating
              dredged material to kill anaerobic organisms).
2.3           Dredged Material Discharge for Beneficial Uses

This manual  also covers testing procedures for non-conventional dredged material discharge practices
including wetlands creation and other beneficial uses of dredged material. Dredged material in these
circumstances should be tested in accordance with this manual as appropriate. For some beneficial use
applications,  other environmental evaluations may be appropriate.   For instance, in some cases site-
specific plant bioassays (not included in this national manual) may be appropriate in addition to or in lieu
of the animal tests (USACE/EPA, 1992).

The potential effects of a discharge of dredged material into waters of the United States may range from
unmeasured to substantial (e.g., EPA,  1987a; Engler et al.,  1991b; Adams et al., 1992). However, the
vast majority  of dredged material is not sufficiently contaminated to prevent its potential use for discharge
in beneficial use applications such as construction of wetlands, oyster reefs, submerged aquatic vegetation
beds, or other purposes. Dredged material used for such applications  generally will not require rigorous
testing or evaluation. The focus is generally on compliance  with other aspects of the  Guidelines.
However, if there is a reason to believe that dredged material proposed for discharge at, for instance, a
beneficial use site contains contaminants, the evaluation of contaminant effects contained in this manual
is applicable.

This manual  applies to dredged material used for beach nourishment.  Beach nourishment normally
involves hydraulic or mechanical placement of uncontaminated materials on or near a shoreline. Dredged
material proposed for beach  nourishment is usually excluded from chemical or biological testing; the
focus is on analysis to  determine physical compatibility  as measured by grain  size and total organic
carbon (see Section 9.1). However, if mere is a reason to believe that contaminants are present, further
evaluation should be performed.
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2.4            The Role of Biological Evaluations (Toxicity and/or Bioaccumulation Tests) in the
               Manual

As noted in Section 230.61 of the Guidelines, the evaluation process will usually entail investigation of
potential biological effects, rather than merely chemical presence, of the possible contaminants. Biological
evaluations serve to integrate the chemical and biological interactions of the suite of contaminants which
may be  present in a dredged material sample,  including their availability for biological uptake, by
measuring their effects on test organisms. Within the constraints of experimental conditions and the end-
points of effects measured, biological evaluations provide for a quantitative comparison of the potential
effects of a dredged material when compared to reference sediments. Thus, a specified level of change
compared to reference conditions and a statistically significant result in this comparison indicate that the
discharge of the dredged material in question may cause a direct and specific biological effect under test
conditions and, therefore, has the potential to cause an ecologically undesirable impact. Guidance for the
conduct of biological tests is given in Sections 11 and 12.

Dredged material potentially contains a myriad of chemical contaminants  which may adversely impact
aquatic organisms.  The literature is replete with examples where aquatic organism sensitivity varies with
the type  of contaminant (e.g., see Rand and Petrocelli, 1985) and, as a result, a suite of aquatic species
are routinely recommended to fully assess the impact of contaminants on a biological community. In this
manual, three sensitive species are recommended for the water column and whole sediment toxicity tests.
In the case of the latter, two species can  be used, provided they cover three functional characteristics:
filter feeder, deposit feeder, burrower. In both cases, at least one of these species must be a sensitive
"benchmark" species. For assessing bioaccumulation, adequate  tissue  biomass and the ability to ingest
sediments is more important than taxon sensitivity. Where possible, two species should be used to assess
potential bioaccumulation unless adequate regional data are available to justify single species testing.

It is important to recognize that dredged material bioassays (toxicity and  bioaccumulation tests) are subject
to interpretation and are not precise predictors of environmental effects. This manual does not provide
quantitative guidance  on interpreting the ecological meaning  of  such  effects (e.g.,  the  ecological
consequences of a given tissue concentration of a bioaccumulated contaminant or the consequences of that
body burden to the animal). Rather, the manual considers statistically significant increases above certain
levels compared to the reference sediment as potentially undesirable. Because a statistically significant
difference is not a quantitative prediction that an ecologically important impact would occur in the field
or vice versa, this manual discusses additional factors to be weighed in evaluating potential ecological
impact. This is more likely to result in environmentally sound evaluations than is reliance on statistical
significance alone.
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Bioaccumulation evaluations indicate biological availability of contaminants in dredged material, which
may bioaccumulate and bioconcentrate in (or, for a few chemicals, biomagnify up) aquatic food webs to
levels which might be harmful to consumers, including human beings, without killing the intermediate
organisms. To use bioaccumulation data, it is necessary to predict whether there will be a cause-and-effect
relationship between the animal's exposure to dredged material and a meaningful adverse elevation of
body burden of contaminants above that of similar animals not exposed to the dredged material.
2.5            The Role of Water and Sediment Chemical Evaluations in the Manual

Chemical evaluations of water and sediments are conducted for the following reasons:

       •      to determine contaminant concentrations in the dredged material

       •      to determine contaminant concentrations in the discharge or reference sites

       •      to determine compliance with water quality standards (WQS).

Chemical evaluations may be made on the basis of previous chemical inventories, when there is a reason
to believe that the dredged material contains no new contaminants, or that there is no difference between
contaminants  in the dredged  material and  the  disposal site [Tier  I;  Section 230.60(a)-(c)  of the
Guidelines]. The latter may be the case where the discharge site is adjacent to the dredging site, and
potential differences  in contaminant bioavailability are considered unlikely.  There may, however, be
concern with potential water column effects which would warrant evaluation of such potential effects (Tier
II; Section 2.6). In  particular, it must be shown that unacceptable levels of dissolved  and suspended
contaminants from the discharge either will not be released and transported to less contaminated areas,
or can be managed.

Initial evaluation of water column chemistry may be carried out through the use of a numerical dispersion
model based on bulk sediment chemistry (Section 5.1.1). If this model  indicates the potential for adverse
effects, a chemical evaluation of potential water column effects may be conducted through the use of elu-
triate tests [Tier II; Section 230.61(b)(2) of the Guidelines]. In this procedure an aqueous extract (i.e.,
an elutriate) is prepared from the material to be discharged, and the dissolved contaminants are compared
to water quality standards with consideration of mixing.  This comparison  requires  that  dissolved
contaminants in reference water (ambient condition) also be analyzed.
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The above elutriate test is used to determine compliance with WQS with consideration of mixing. The
elutriate test provides an indirect evaluation of potential biological effects, because WQS are derived from
toxicity tests of solutions of various contaminants. Even if WQS are met, biological evaluations (see
Section 2.4) must be considered.
2.6            Water Column Effects

The dredged material impact in the water column must be within the available WQS for all contaminants
of concern outside of the mixing zone. If disposal operations result in long-term exposures, compliance
with chronic aquatic and/or human health standards should be evaluated. Wildlife standards, if available,
should also be considered. Water column toxicity tests  are used to provide information on the toxicity
of contaminants not included in water quality standards, and also to indicate possible interactive effects
of multiple contaminants.
2.7            Mixing

Appendix C describes the method to be used for estimating the effect of mixing for water column
evaluations. 40 CFR 230.11(0(2) describes the factors to be considered in defining mixing zones; States
may use additional factors in such definition. This method  is applied in evaluating the potential for
impacts of the portion of dredged material that remains in the water column; all water quality and water
column toxicity data must be interpreted in light of mixing  [Section 230.61 (b)(2)(ii) of the Guidelines].
This is necessary because biological effects (which are  the basis for WQS) are a function of the
biologically available contaminant concentration and exposure time of the organisms. Laboratory toxicity
tests expose organisms to specific concentrations  for fixed periods of time,  whereas in the field  both
concentration and exposure time to contaminants change continuously due to mixing and dilution.  Both
factors interact to control the degree of biological impact. Thus, it is necessary to incorporate the mixing
expected at the discharge site into the interpretation of data.
2.8            Benthic Effects

Generally, the  greatest potential for environmental effects from dredged material discharge lies in the
benthic environment. Deposited dredged material is not mixed and dispersed as rapidly or as greatly as
the portion of the material that may remain in the water column, and bottom dwelling animals living and
feeding on deposited material for extended periods represent the most likely pathways by which adverse
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effects to aquatic biota can occur. Therefore, the major evaluative effort must be placed on deposited
material and the benthic environment, unless there is a compelling reason to do otherwise. The approach
in this manual is  conservative (i.e., protective)  as  it uses whole-sediment bioassays (toxicity and
bioaccumulation tests) to evaluate the solid phase of the dredged material. Sediment chemical analyses
currently cannot be used to directly evaluate the biological effects of any contaminants which may be
present in dredged  material because such potential effects are a function of bioavailability. However, as
noted in Section 2.5, there are circumstances where it may be reasonably assumed that bioavailability in
the dredged material and the discharge site are similar.  When decisions cannot be made using evaluations
in Section  230.60  of the Guidelines, bioaccumulation tests should be used to directly  determine the
bioavailability of potential contaminants.
2.9           Management Options

Some dredged material evaluated in accordance with technical procedures in this manual may demonstrate
a potential for unacceptable environmental impacts or not meet Federally approved State WQS. If so, a
careful case-by-case evaluation of management options (e.g., alternative dredging and discharge methods,
alternative discharge sites, confined disposal, capping, site controls such as covers and/or liners) will be
necessary to determine whether the proposed discharge can be made acceptable or can be brought into
compliance with the Guidelines and State WQS. As previously noted, it is beyond the scope of this
manual to determine whether  a material which would not otherwise comply with the Guidelines, could
be brought into compliance through appropriate management actions or other discharge methods.
2.10          The Relationship of the Inland Testing Manual  to  Other USACE/EPA Dredged
              Material Management Efforts

2.10.1        Relationship of the Manual to the USACE/EPA Framework Document

EPA and USAGE have long recognized the need for a consistent technical framework for decision-making
regarding the discharge  of dredged material  in ocean,  near  coastal,  and inland waters (e.g., see
Francingues et al., 1985; Wright and Saunders, 1990). This manual  is one of a series of guidance
documents jointly  developed by  EPA and the USAGE in response to that recognition. This series of
guidance documents includes the "Evaluating Environmental Effects of Dredged Material Management
Alternatives - A Technical Framework" (USACE/EPA, 1992) which articulates those factors (including
the potential for  and degree of contaminant-related impacts) to  be considered in identifying the
environmental effects of dredged material management alternatives on a continuum from uplands to
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oceans, and which meet the substantive and"procedural requirements of NEPA, CWA and MPRSA. The
companion testing manual for ocean disposal, the Green Book (EPA/USACE, 1991) is included in the
series. Application of the testing guidance in this manual within the context of the Framework Document
will allow for consistency in decision-making with respect to technical considerations, across statutory
boundaries and  with consideration of the continuum of dredged material discharge options.
2.10.2        Relationship of the Manual to the EPA/USACE Green Book

Although the Ocean Dumping and the CWA programs carry out their functions under different mandates
and different environments (estuarine, lake and riverine versus ocean), there is a considerable overlap in
terms of practical application. The Guidelines are statutorily directed to be based upon criteria comparable
to those developed under Section 403(c) for the territorial seas, contiguous zone, and ocean. Additionally,
in previous guidance both EPA and USACE have acknowledged the ecological similarity of all aquatic
areas and the need for a consistent technological analysis framework, particularly when the waters of the
United States under consideration for a discharge are near-coastal. While  details  of this manual are
necessarily different from one addressing only ocean waters, the tiered testing framework and concepts
of the Green Book are an appropriate paradigm. The Inland Testing Manual also utilizes  the  Green
Book's reference site approach which provides a more accurate data base for cumulative impact analysis.

Dredged material transported  for purposes of dumping or disposal  seaward of the baseline of the
territorial  sea will continue to be regulated  under the MPRSA (commonly referred to as  the Ocean
Dumping Act). MPRSA-regulated dredged material disposal will be tested in accordance with procedures
outlined in the Green Book (EPA/USACE,  1991). As previously discussed, dredged material used as fill
within the territorial sea, such as  for beach nourishment, is regulated under the CWA and will be tested
in accordance with this manual.
2.10.3        Relationship of the Manual to EPA's Contaminated Sediment Strategy and Sediment
              Quality Criteria

EPA is developing a Contaminated Sediment Management Strategy (Strategy) which is a multi-program
effort to address contaminated aquatic sediments in the United States. The Strategy is intended to improve
the understanding of the extent and severity of sediment contamination and to propose prevention, control,
and remediation programs. The Strategy describes the policy framework and specific actions EPA could
take to  promote the  consideration of and reduction of ecological and human health risks posed by
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sediment contamination. The Strategy also recommends a comprehensive research program and outreach
activities with other agencies and the general public.

One component of the Strategy is the development of Sediment Quality Criteria (SQC), which are derived
numerical values representing the concentration of chemicals  in sediment which are determined to
adversely affect benthic organisms. SQC are included in EPA's approach to defining contamination in
sediments, and  are envisioned to play a range of roles in all programs, from assessment to remediation.
When finalized, SQC likely will be incorporated into the Inland Testing Manual in Tier II. SQC could
also form the basis for State SQS. The Inland Testing Manual is structured such that evolving science
may be readily  merged into the document.
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PART H - EVALUATION OF POTENTIAL ENVIRONMENTAL IMPACT
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3.0            OVERVIEW OF TESTING AND EVALUATION

As noted in Section 1.2.2.1,  conclusions reached  utilizing this manual will be used to  make factual
determinations of the potential effects of a proposed discharge of dredged or fill material on the physical,
chemical and biological components of the aquatic environment. Such factual determinations are used to
make findings of  compliance or noncompliance with relevant parts of Sections 230.10(b)  (including
compliance with  established  water  quality  standards) and 230.10(c)  (determinations  of potential
contaminant-related impacts to aquatic resources).
3.1            Tiered Testing and Evaluation

The tiered approach to testing used in this manual must be initiated at Tier I. It is designed to aid in
generating physical, chemical, toxicity and bioaccumulation information, but not more information than
is necessary to make factual determinations. This allows optimal use of resources by focusing the least
effort on disposal operations where the potential (or lack thereof) for unacceptable adverse impact is
clear, and expending the most effort on operations requiring more extensive investigation to determine
the potential (or lack thereof) for impact. To  achieve this objective, the procedures in this manual are
arranged in a series of tiers, or levels of intensity (and cost) of investigation. Tiered testing results in
environmental protection in the context of more efficient completion of necessary evaluations and reduced
costs, especially to low-risk operations. Disposal operations that obviously have low environmental impact
generally should not  require  intensive investigation  to  make factual determinations.  Evaluation at
successive tiers is based on more extensive and specific information about the potential impact of the
dredged material, that may be more time-consuming and expensive to generate, but that allows more and
more comprehensive evaluations of the potential for environmental effects.  At any tier except for Tier
IV, failure to satisfactorily determine the potential for unacceptable aquatic environmental impact, or to
develop sufficient information to make factual determinations, results in additional testing at a subsequent,
more complex tier unless a decision is made  to seek other disposal alternatives (thereby avoiding the
potential for unacceptable aquatic environmental impacts).

It is necessary to proceed through the tiers only until information sufficient to make factual determinations
has been obtained. For example, if the available information is sufficient to make factual determinations,
no further testing is required.

The initial tier (Tier I) uses readily available,  existing information (including all previous testing). For
certain  dredged materials  with readily apparent potential  for environmental impact (or lack thereof),
information collected in Tier I may be sufficient for making  factual determinations.  However, more
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extensive evaluation (Tiers II, III and IV) may be needed for other materials with less clear potential for
impact or for which Tier I information is inadequate.

Tier II is concerned solely with sediment and water chemistry. Tier III is concerned with well-defined,
nationally accepted toxicity and bioaccumulation testing procedures.  Tier IV allows for  case-specific
laboratory and field testing, and is intended for use in unusual circumstances.

The approach is to enter Tier I and proceed as far as necessary to make factual determinations. Although
it is not always necessary that all dredged material be evaluated through all tiers, there must be enough
information available to make determinations on all aspects of the Guidelines relating to water column
impact, benthic toxicity and benthic bioaccumulation. It is acceptable to carry water-column and benthic
evaluations, or toxicity and bioaccumulation evaluations, to  different  tiers to generate the information
necessary and sufficient to make these determinations.

Prior to initiating  testing,  it is essential  that the informational requirements of preceding  tiers be
thoroughly understood and that the information necessary for interpreting results at the advanced tier be
assembled. For example, it is always appropriate to gather all relevant available information and identify
the chemicals of concern for the dredged material  in question even though it may be clear without formal
Tier I evaluation that further assessment will be necessary.

The tests in this manual reflect the present state-of-the-art procedures for dredged material evaluation.
However, it is recognized that the evaluation of dredged material is an evolving field. It is anticipated
that, as new  methods of evaluation are developed and accepted, they  will be integrated  into the tiered
framework. The tiered approach will be maintained because of the efficiency afforded by its hierarchical
design.

The tiered approach used in the manual is summarized in Figure 1,  and  additional detail on water column
and benthic evaluation is presented in Figures 2 and 3. These flowcharts should be used  in conjunction
with a careful reading of the corresponding guidance presented in  this  manual,  in particular Sections 4,
5, 6 and 7. The sections or figures  in  the manual that present  the technical  guidance shown by the
flowcharts are indicated in the boxes  on the figures.
3.2            Control and Reference Sediments

It is important to clearly distinguish between control and reference sediments in the context of testing for
benthic impacts. In general, control sediment is that within which the organisms resided prior to
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                                                                      37
                                                                                                                TIER H
                                                                     FROM FIGURE 1
                                                             EVALUATE POTENTIAL WATER-COLUMN
                                                                        IMPACT
                                                                                     WATER-COLUMN TOXIOTY
                                     WQS SCREEN,
                                    MODEL ASSUMED
                               TOTAL RELEASE OF SEDIMENT
                                 CONTAMINANTS TO THE
                                    WATER COLUMN
                                       (10.1.1)
                                                                MEASURE DISSOLVED
                                                           CONCENTRATIONS OF CONTAMINANTS
                                                             OF CONCERN m WATER COLUMN
                                                                     (10.1.2)
                                                                 MODEL DISSOLVED
                                                          CONCENTRATIONS OF CONTAMINANTS
                                                            OF CONCERN IN WATER COLUMN
                                                                     (10
                                             MEASURE TOMCITY
                                             OF DM SUSPENSION
                                                   (111)
                                               TO»CITY>10*
                                              DIFFERENCE AND
                                           SIGNIFICANTLY DIFFERENT
                                               THAN DILUENT
                                                 WATER?
                                                MODEL DM
                                             SUSPENDED PHASE
                                             IN WATER COLUMN
                                                  11.1.7)
                        WATER-COLUMN
                           TOMCITY
                                                                                                               UNUSUAL
                                                                                                            CIRCUMSTANCE
                                                        INSUFFICIENT INFORMATION
                                                                       CONDUCT CASE-SPECIFIC
                                                                          TOMCITY TESTS
                                                                              (11.4)
                           KEY TO NOMENCLATURE

                    DM  DREDGED MATERIAL
                   Was  WATER QUALITY STANDARDS
                                                       DM NOT
                                                      PREDICTED
                                                     TO RESULT IN
                                                    WATER-COLUMN
                                                       TOMCITY
      ARE
  CASE-SPECIFIC
CRITERIA MET AFTER
  INITIAL MIXING?
     DM
  PREDICTED
 TO RESULT IN
WATER-COLUMN
   TOXCITY
                 TIER  IE
     LETHAL CONCENTRATION TO SOX OF
     TEST ORGANISMS. EQUAL TO
     ACUTE TOXIdTY CONCENTRATION

EC jo EFFECTS CONCENTRATION. EQUIVALENT
     TO LC so FOR NON-LETHAL ACUTE EFFECTS
TIER3Z
Figure 2.        Illustration of Tiered Approach to Evaluating Potential Water Column Impacts of Dredged
                     Material.
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                                                                     38
                                                             FROM FIGURE 1
                                                       EVALUATE POTENTIAL BENTHIC
                                                                IMPACT
                                                                                    BENTHIC BIOACCUMULATION
                                                                            CALCULATE THEORETICAL
                                                                           BIOACCUMULATION POTENTIAL
                                                                                     (10.2)
                                                           DM NOT
                                                          PREDICTED
                                                         TO RESULT IN
                                                           BENTHIC
                                                          BIOACCUMU-
                                                            LATION
                                                           OFNPO
                                                                                      OTHER
                                                                                   CONTAMINANTS
                                                                                    OF CONCERN
                                                                  DM
                                                              EXCEEDS REF?
                                                                  (5.2)
                                                                            MEASURE BIOAVAILABILITY
                                                                                    (12.1)
                          MEASURE TOXICITY
                               (11.2)
                           DM
                        PREDICTED
                       TO RESULT IN
                          ACUTE
                         BENTHIC
                         TOXICITY
                                DM>
                               REF BY
                        MORE THAN ALLOWABLE
                            PERCENTAGE?
                                (6.2)
                                                 DM
                                               PREDICTED
                                               TO RESULT
                                                IN ACUTE
                                                BENTHIC
                                                TOXICITY
                                              DM
                                           PREDICTED
                                          TO RESULT IN
                                            BENTHIC
                                           BIOACCUMU-
                                             LATION
                                                                                            DM NOT
                                                                                          PREDICTED
                                                                                         TO RESULT IN
                                                                                            BENTHIC
                                                                                          BIOACCUMU-
                                                                                            LATION
    ARE
CASE-SPECIFIC
CRITERIA MET?
     .3)
                                                                                       INSUFFICIENT
                                                                                       INFORMATION
        UNUSUAL
     CIRCUMSTANCES
                                         CONDUCT CASE-SPECIFIC
                                             TOXICITY TESTS
                                                 (11.4)
                                                        MEASURE EMPIRICAL STEADY
                                                          STATE BIOACCUMULATION
                                                                 (12.2)
                           DM NOT
                         PREDICTED
                        TO RESULT IN
                           BENTHIC
                          TOXIOTY
                                ARE
                           CASE-SPECIFIC
                           CRITERIA MET?
                                 .1
                                                  DM
                                               PREDICTED
                                              TO RESULT IN
                                                BENTHIC
                                                TOXICITY
KEY TO NOMENCLATURE
                                                                                    DM
                                                                               FIELD ORGANISMS?
                                                                                    (7.2)
                                                                DM
                                                             PREDICTED
                                                            TO RESULT IN
                                                              BENTHIC
                                                             BIOACCUMU-
                                                               LATION
                         DM NOT
                        PREDICTED
                       TO RESULT IN
                         BENTHIC
                        BIOACCUMU-
                         LATION
                                                                  ARE
                                                              CASE-SPECIFIC
                                                              CRITERIA MET?
                                                                   .2)
                                                                                                   TIER  I
                                                                                                         TIER  IK
                                                                                                           UNUSUAL
                                                                                                        CIRCUMSTANCES
                                                                                                   TIER  1Z
 DM  DREDGED MATERIAL

REF  REFERENCE SEDIMENT

NPO  MOM POLAR ORGANICS

 >•  STATISTICALLY GREATER THAN

FDA  USFDA ACTION LEVELS
     FOR POISONOUS AND
     DELETERIOUS SUBSTANCES
     IN FISH AND SHELLFISH
     FOR HUMAN FOOD
1        Figure 3.        Dlustration of Tiered  Approach to Evaluating Potential Benthic  Impacts of Deposited
2                           Dredged Material.
                                                                                                                 DRAFT

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	39	

collection in the field or is that within which they were cultured in the laboratory, and serves to confirm
the health of the test animals and the acceptability of the test conditions. Generic control sediments are
also possible and consist of field-collected or laboratory prepared sediment. Reference sediment is the key
to the evaluation of dredged material. Results of tests using reference sediment provide the point of
comparison (reference point) to which benthic effects  of dredged material are compared.

In some cases, it may be appropriate to use more than one reference sediment for a single  dredging
project. This could occur when the dredged material or the disposal site has a wide range of grain-sizes,
when management needs suggest that disposal of different dredged materials at different locations in the
disposal site is desirable, or when discharge at more than one site is being considered. One reference site
can serve more than one disposal site.
3.2.1          Reference Sediment Sampling

Reference sediment fNOTE: The  reference sediment concept is  the subject of a CWA Section 404
rulemaking under development.  The rule will be proposed in the  Federal Register in 1994 for public
comment and will be promulgated prior to issuance of the final Inland Testing Manual.] is generally
collected outside the influence of previous disposal operations at a dredged material disposal site, but near
enough to the  disposal site that the reference sediment is subject to all die same influences (except
previously disposed dredged material) as the disposal site. If there  is a potential for sediment migration
or there is a reason to believe that previously disposed sediment has migrated, reference sediment should
be collected from an area outside the disposal site that is not expected to be influenced by material from
the site. There  are four potential reference sampling approaches  as discussed below.

Reference Point Approach: This approach  is used  when the disposal site is known to be sufficiently
homogeneous that a single reference location is representative of the disposal site. A single reference
location is sampled and the sediment is tested concurrently with the dredged material. The bioassay results
from the reference sediment are  statistically compared to those  obtained from benthic toxicity and
bioaccumulation tests of the material to be dredged.

Reference Area Approach: This approach is used when the disposal site is known to be heterogeneous and
more than one  reference location must be sampled to  adequately characterize the disposal site.  Several
reference locations are sampled and a composite of all of the sediments are tested concurrently with the
dredged material. The bioassay results from the reference sediment composite are statistically compared
to those obtained from benthic toxicity and bioaccumulation tests of the material to be dredged.
                                                                                 DRAFT

-------
	40	

Periodic Reference Point Approach: This approach could, theoretically, be used when it is not desirable
or possible to sample the reference location each time that dredged material is to be tested. Values from
the homogeneous reference location collected over a period of time are used to develop decision guidance
values which are statistically compared to those obtained from benthic toxicity and bioaccumulation tests
of the material  to be dredged.

Periodic Reference Area Approach: This approach could, theoretically, be used when it is not desirable
or possible to sample the heterogeneous reference locations each time that dredged material is  to be
tested. Values from heterogeneous reference locations collected over a period of time are used to develop
decision guidance values which are statistically  compared to those obtained from benthic toxicity and
bioaccumulation tests of the material to be dredged.

Appendix D, Statistical Methods, provides  guidance for  conducting statistical comparisons  for the
reference point and reference area approaches. It does not provide guidance for the  use of either of the
"periodic"  approaches.  Although the Ocean Testing  Manual (EPA/USACE,  1991)  suggests that a
"periodic" approach may be used, subsequent scrutiny has shown that the conditions and assumptions
required for either of the "periodic" approaches can very rarely, if ever, be met. Hence, the use of these
approaches is strongly discouraged and is not discussed further in this manual.
3.2.2          Reference Sediment Sampling Plan

The importance of thoughtful selection of the reference sampling approach cannot be overemphasized.
To ensure that an appropriate approach is used, information gathered during the site specification process
or other studies should be consulted for both the disposal and the reference sites. In some instances there
are differences in the statistical methods used in comparing results from the various reference sampling
methods to those obtained from the dredged material being evaluated. There may also be differences in
costs among the approaches. Prior to selecting an approach, it is imperative that Appendix D be consulted
to determine which approach best fits specific concerns and conditions, including feasibility, technical
validity, and cost.

A well-designed sampling plan is essential to the collection, preservation, and storage of samples so that
potential toxicity and bioaccumulation can be accurately assessed (Section 8). The implementation of such
a plan is equally essential for dredged material, control sediment, and reference sediment.
                                                                                 DRAFT

-------
41
                      DRAFT

-------
                                     42
                  EVALUATE EX&TJN(3
    WATER COLUMN
      BENTHOS
    MEASURE AND
  MODEL DISSOLVED
   CONTAMINANTS;
  COMPARE TO WQS
CALCULATE THEORETICAL
   BIOACCUMULATION
  POTENTIAL; COMPARE
    TO REFERENCE
 MEASURE TOXICITY;
 MODELSUSPENDED
 PHASE; DETERMINE
TOXICITY AFTER MIXING
  MEASURE TOXICITY;
      MEASURE
  BIOACCUMULATION;
 COMPARE TO FDA LIMITS
  AND TO REFERENCE
     CONDUCT
   CASE-SPECIFIC
   TOXICITY TESTS
      CONDUCT
    CASE-SPECIFIC
      TOXICITY;
   BIOACCUMULATION;
     OTHER TESTS
                                                                  TJE8 I
                                                                  dVireP
                                                            EXISTS ^FORMATION)
     TIER II
(SOLELY CONCERNED
 WITH CHEMISTRY)
     TIER III
(GENERIC BIOASSAY
   [TOXICITY AND
 BIOACCUMULATION]
      TESTS)
    TIER IV
(SPECIFIC BIOASSAY
   [TOXICITY AND
 BIOACCUMULATION]
 AND OTHER TESTS)
                                                                    DRAFT

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	43	

4.0            TIER I EVALUATION

One of the purposes of Tier I is to determine whether factual determinations can be made on the basis
of existing  information.  Tier I  is  a comprehensive analysis of all existing and readily  available,
assembled,  and interpreted information on  the proposed dredging project,  including all  previously
collected physical, chemical, and biological monitoring data and testing for both the dredged material
excavation site and the proposed disposal site. Only limited testing, to determine the applicability of
exclusions, may be necessary in this tier.

If the information set compiled in Tier I is adequate to meet the exclusions or is complete and comparable
to that which would satisfy Tier II, III, or IV, as appropriate, factual determinations can be made without
proceeding into the higher tiers (Figure 1). For an evaluation to be completed within Tier  I,  the burden
of evidence  of the collected information must be adequate to make factual determinations.

The initial focus of the Tier I evaluation is on information relevant to Sections 230.60 (a), (b), (c), and
(d) of the Guidelines and the potential for contaminant-associated impacts upon discharge. These four
sections of the Guidelines fully define the exclusions from testing, which are summarized  below.

If an evaluation of the dredging site indicates that the dredged material is not a "carrier of contaminants",
testing may not be necessary. Such situations  are most likely to arise if:   the dredged material is
composed primarily of sand, gravel and/or inert materials; the sediments are from locations far removed
from sources of contaminants; the sediments are from depths deposited  in preindustrial times and  not
exposed to modern sources of pollution. However, potential  impacts from natural mineral deposits must
also be considered.

Testing may also not be necessary "where the discharge site is adjacent to the excavation site and subject
to the same sources of contaminants, and materials at the two sites are substantially similar "(Section
230.60 (c)).  However, some physical and chemical testing may be necessary to confirm that the two sites
are "substantially similar". The rationale behind this exclusion from testing is that when 1)  the discharge
and  excavation sites  are adjacent,  2) the concentration of contaminants  in the  two  sites  are not
substantially different, and 3) the geochemical  environments are similar, then the bioavailability  of
contaminants at the two sites are likely to be similar.  This  exclusion can apply even if the dredged
material is a carrier of contaminants, providing that "dissolved materials and suspended particulates can
be controlled to prevent carrying pollutants to less contaminated areas".

Section 230.60 (d) states that testing may not be necessary with material likely to be a carrier  of
contaminants if constraints acceptable to the USAGE District Engineer and EPA Regional Administrator
                                                                                 DRAFT

-------
	44	

are available to "reduce  contamination  to  acceptable levels  within the disposal site and to  prevent
contaminants from being transported beyond the boundaries of the disposal site". Such constraints may
involve technologies such as capping and underwater containment. Design and monitoring requirements
for such constraints should be determined by the Regional Administrator and District Engineer on a case-
by-case basis.

If the exclusionary criteria are satisfied, factual determinations  for the dredged material can be made and
no further evaluation is necessary. If the exclusionary criteria are not met, the material is evaluated based
on all existing information. This information should include chemical  information  and, if appropriate,
existing data on the toxicity and bioaccumulation potential of the dredged material and of the reference
sediment. The information must be sufficient to determine if water quality standards are met and, if
appropriate, whether  1 % of the LCjo or ECjo of each tested species will or will not be exceeded in the
water column following mixing. If adequate information is not available for a Tier I  evaluation, the
process moves to Tier II.

Even if factual  determinations cannot be made  on the basis of Tier I information,  the information
collected can be put to use in later tier analyses. Another purpose of Tier I is to identify the contaminants
of concern (if any) in the dredged material.  This information  is used to select analyses  in Tiers II, III,
and  IV. Similarly, other information collected  in Tier I may be used to satisfy all  or portions of
evaluations in other tiers. It is necessary to  proceed through the tiers only until a factual determination
is reached. Rigorous information collection and assessment in Tier I inevitably saves time and resources
in making  final determinations.

Annual or  episodic dredging,  undertaken to maintain existing  navigation improvements, may warrant a
periodic Tier I  reevaluation.  The  general recommendation of EPA and USAGE is that the  interval
between reevaluation of Tier I data for these projects not exceed three years or  the dredging cycle,
whichever  is longest.  If there is reason to believe that conditions have changed, then the time interval for
reevaluation  may be less than three years. As a minimum, this reevaluation should include a technical
reassessment of all new and previously evaluated  physical, chemical  and biological data, changes in
sediment composition or deposition (e.g.,  industrial development in the watershed), improvements in
analytical methods and contaminant detectability, quality assurance considerations and any regulatory
changes.
                                                                                  DRAFT

-------
	45	

4.1           Compilation of Existing Information

The potential for contaminants  to  have been  introduced  to  the dredged  material, evaluated with
consideration of the physical nature of the dredged material, and the proposed disposal site, allows case-
by-case determinations  of whether the proposed discharge  of dredged  material may  result in
contamination, bioaccumulation or toxicity above reference levels.  Section 230.60 (b) of the Guidelines
lists a number of factors which should be considered when evaluating the potential for contamination at
the dredging (i.e., extraction) site. These  factors represent sources of contamination, pathways of
contaminant transport, and naturally occurring substances which may be harmful to aquatic biota:

       •      urban and agricultural runoff
       •      sewer overflows/bypassing
       •      industrial and municipal wastewater discharges
       •      previous dredged or fill discharges
       •      landfill leachate/groundwater discharge
       •      spills of oil or chemicals
       •      releases from Superfund and other hazardous waste sites
       •      illegal discharges
       •      air deposition
       •      biological production (detritus)
       •      mineral deposits.

The information gathering phase of Tier I evaluations has to be as complete  as is reasonably possible,
including existing information from all reasonably available sources. This will increase the likelihood that
determinations concerning the impact of dredged material  may be made at initial tiers.  Sources of
available information include the following,  without limitation:

       •      Results of prior physical, chemical,  and biological  tests and monitoring of the material
              proposed to be disposed.
       •      Information describing the source of the material to be disposed which would be relevant
              to the identification of potential contaminants of concern.
       •      Existing data contained in files of agencies such as EPA or USAGE or otherwise available
              from public or private sources.  Examples of sources from which relevant information
              might be obtained include:
              •        Selected Chemical Spill Listing (EPA)
              •        Pesticide Spill Reporting System  (EPA)
              •        Pollution Incident Reporting System  (United States Coast Guard)
                                                                                 DRAFT

-------
	46	

              •       Identification of In-Place Pollutants and Priorities for Removal (EPA)
              •       Hazardous waste sites and management facilities reports (EPA)
              •       USAGE studies of sediment pollution and sediments
              •       Federal STORET,  BIOS, CETIS, and ODES databases (EPA)
              •       Water and sediment data on major tributaries (Geological Survey)
              •       NPDES permit records
              •       Agencies with contaminant or related information, for instance, Fish and Wildlife
                      Service (FWS), National Oceanic and  Atmospheric Administration  (NOAA),
                      regional planning commissions, state resource/survey agencies
              •       CWA 404(b)(l) evaluations
              •       Pertinent and applicable research reports
              •       MPRSA  103  evaluations
              •       Port and  marina authorities
              •       Colleges/Universities
              •       Records of State agencies, (e.g., environmental,  water survey, transportation,
                      health)
              •       Superfund sites, hazardous waste sites
              •       Published scientific literature.

Sources may contribute differing types and quantities of contaminants to sediments. For example, a matrix
of potential correlations between industrial sources and specific contaminants is provided in Table 1. This
matrix is, however, not all inclusive and makes  no accounting  for current pollution control practices.

There are also a number of factors which influence the pathways between contaminant sources and the
dredging and disposal sites, including:

       •      bathymetry
       •      water current patterns
       •      tributary flows
       •      watershed hydrology and land uses
       •      sediment and soil types
       •      sediment deposition rates.

More detailed site-specific guidance  for reaching  administrative decisions concerning the impact  of a
dredged  material discharge may  be  developed  by particular EPA Regions and USAGE Districts by
considering available scientific information and locally important concerns. In evaluating the likelihood
                                                                                DRAFT

-------
8
ON
1
Boat Refueling
Boat Manufacturing/
Boat Repair
Potential non-point
Sources
Waste Water
Treatment Plants
Valuable Mineral
Mining
Utilities
Textiles
SulfurlcAcId
Steel/Iron
Steam Power
Rubber
Pulp and Paper Mills
Printing Plates
Plastics
Pigments/Inks
Photographic
Phosphorus
Phosphate Mining
Petroleum Refining
Peatlcldes/Fertil ttere
Perfume
Oxide Manufacturing
NHricAcW
Manufacturing
Metallurgical Processes
Metal Finishing Refining
Meat Products
Leather/Tanning
Fruits and Vegetables
Rat Glass
Explosives
Electrical
Dye
Detergents /Surfactants
Dairy
Corrosion Metallurgy
. Commercial Farming
Chemical Manufacturing
Batteries
Automobile
Anti-FoulIng Paints
Ammunitions
Aluminum
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	48	

that discharge of a dredged material may cause contaminant associated impacts, concern decreases with
the increase of factors such as:

       •      isolation  of  the dredging operation from  known existing  and historical sources of
              contamination
       •      time since historical sources of contamination have been remediated
       •      number and frequency of maintenance dredging operations since abatement of the source
              of contamination
       •      mixing and dilution occurring between the contamination source and the dredging site
       •      transport and potential deposition of sediment in the dredging area from sources other
              than those potentially affected by contamination
       •      grain size of the dredged material.

Concern regarding contaminant associated impacts increases with the increase of factors  such as the
number,  amount, and lexicological importance of contaminants:

       •      known to have been introduced to the dredging  site
       •      suspected to  have been introduced to the dredging site
       •      included in continuing input from existing sources
       •      included in historical sources.

These and other considerations are complexly interrelated; i.e., the acceptable degree of isolation from
sources of  contamination depends  on the  number,  amount,  and toxicological importance of the
contaminants as well  as on all other factors. These considerations have to be evaluated for all dredged
material. Even so,  it is desirable that local guidance be developed, based on technical evaluations, that
describes the emphasis on factors deemed appropriate  in each area. In all cases, the decisions that are
based on these factors must  be compatible with the Guidelines.
4.2            Identification of Contaminants of Concern

In the Tier I decision sequence (Figure 1), the first possibility is that more information is required to
make a factual determination.  A critical prerequisite to generating this information and one which is
crucial to the success of the testing program is deciding, on a case-by-case basis, which contaminants are
of concern, particularly for 401 certification, in the dredged material being evaluated. To determine the
contaminants  of concern,  it may be  necessary to  supplement available information with additional
chemical analyses of the dredged material.  Contaminants of concern are not restricted to compounds
                                                                                 DRAFT

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	49	

which inhibit organisms but also those which promote undesirable organisms or growth (e.g., nutrients
such as phosphorous - Nakaniski et al., 1986). However note that in at least some cases nutrient releases
may be minimal and of no environmental concern (e.g., Tavolaro and Mansky,  1985).
4.2.1         Microbial Contamination

As noted hi Section 2.2, this manual only addresses microbiological  concerns to the extent that they
address State 401 certification requirements. To this end, major areas of concern and pertinent sources
of information addressing these and other relevant microbiological issues are provided below.

If sediments are suspected to have high levels of microbial contamination and dredging or disposal sites
are close to shellfish beds, swimming beaches or drinking water intakes, then microbial sediment analyses
may be required. Useful references include: EPA (1978); Gerba et al. (1979); Dutka et  al.  (1988) and
Helmer et al. (1991). Appropriate state health and water quality agencies should be consulted for guidance
and appropriate methods for measuring microbial contamination.

There are three major areas of concern for microbiological contamination and  effects related to dredged
sediments:  (1) contamination of harvestable shellfish (e.g., Hood et al., 1983; Bruckhardt et al.,  1992;
Martinez-Manzanares et al.,  1992); (2)  body contact, generally related  to swimming beaches  (e.g.,
Fleisher, 1991; Helmer et al., 1991); (3) contamination of drinking water (e.g., Geldreich, 1991; Helmer
et al.,  1991). As noted in the Guidelines (e.g., 230.21, Suspended Particulates, and elsewhere), the
ultimate concern is that "...pathogens and viruses...may be biologically available".

Sediments generally  contain higher concentrations of indicators of fecal contamination and  pathogens,
such as Salmonella and viruses,  than occur in the water column (e.g., Chen et al.,  1979; Gerba et al.,
1979; LaBelle et al., 1980). Further, these microorganisms survive longer in  the sediments  than in the
water column (e.g., DeFlora et al., 1975; Smith et al.,  1978; Borrego et al., 1983; Rao et  al., 1984).
Sediments  have  been shown  to  be a source of microorganisms released to  the water  column  (e.g.,
VanDonsel and Geldreich, 1971; Shiharis et al., 1987; Hardina and Fujioka, 1991). More specifically,
dredging and disposal have been  shown to release these microorganisms (e.g., Grimes, 1975; Babinchak
et al.,  1977).
                                                                                DRAFT

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	50	

4.2.2          Chemical Contamination

Nationally,  it  is  difficult to  specify  a single set  of contaminants  that  adequately addresses all
environmental concerns. However, regions may develop their own general contaminants of concern list
for routine permitting purposes. In some dredged materials, there may be no contaminants of concern.
Different disposal operations may  have their own  set of contaminants of environmental concern that
should be adequately evaluated for each operation.

Identifying specific contaminants that are of concern in a particular dredged material is dependent on the
information collected for Tier I. In some instances, it may be sufficient to perform confirmatory analyses
for specific contaminants of concern identified in Tier I. In other cases, where the initial evaluation
indicates that a variety of contaminants of concern may be present, chemical analysis of the dredged
material  could provide a useful inventory, and bulk sediment chemistry analysis conducted according to
the guidance in Section 9.3 may be appropriate and, in  fact,  would be necessary to conduct the Tier II
water quality screen and the theoretical bioaccumulation potential determination. Contaminants always
of interest, if present, are those for which there are  FDA limits or state fish advisories and where WQS
exceedances exist. Other contaminants that should be included are those that might reasonably be expected
to cause  an  unacceptable adverse impact if the dredged material is discharged.

The contaminants of concern in each dredged material should be identified on the basis of the following,
keeping in mind the discussion in Sections 9.3, 9.4, and 9.5:

       •      presence in the  dredged material
       •      presence in the  dredged material relative to the concentration in the reference sediment
       •      lexicological importance
       •      persistence in the environment
       •      propensity to bioaccumulate from sediments.
               The major chemical properties controlling the propensity to bioaccumulate are:

                       Hydrophobicity
                       Literally,  "fear of water"; the property of neutral  (i.e., uncharged) organic
                       molecules that causes them  to associate with surfaces or  organic solvents rather
                       than to be in aqueous  solution. The presence of a neutral surface  such as an
                       uncharged organic molecule causes water molecules to become structured around
                       the intruding entity. This structuring is energetically unfavorable, and the neutral
                       organic molecule tends to  be partitioned to  a less energetic phase if one is
                       available. In an operational sense, hydrophobicity is the reverse of aqueous solu-
                                                                                 DRAFT

-------
	51	

 bility.  The octanol/water partition  coefficient (K^, log K^, or log P)  is a
 measure of hydrophobicity. The tendency for organic chemicals to bioaccumulate
 is  related to  their  hydrophobicity.  Bioaccumulation  factors increase with
 increasing hydrophobicity up to  a log K^ of about 6.00. At hydrophobicities
 greater than about log K^ = 6.00, bioaccumulation factors tend not to increase
 due, most likely, to reduced bioavailability.

 Aqueous Solubility
 Chemicals such  as acids, bases,  and salts that speciate (dissociate) as charged
 entities tend to  be water-soluble and those  that do  not speciate (neutral and
 nonpolar organic compounds) tend to be insoluble, or nearly so. Solubility favors
 rapid uptake  of chemicals by organisms, but at the same  time  favors rapid
 elimination, with the result that soluble chemicals generally do not bioaccumulate
 to  a great  extent. The soluble free ions of certain heavy metals are exceptional
 in that they bind  with tissues and thus are actively bioaccumulated by organisms.

 Stability
 For chemicals to bioaccumulate, they must be stable, conservative, and resistant
 to degradation (although some contaminants degrade to other contaminants which
 do bioaccumulate). Organic compounds with structures that protect them from
 the catalytic  action  of enzymes or from  nonenzymatic hydrolysis tend to
 bioaccumulate. Phosphate ester pesticides do not bioaccumulate because they are
 easily hydrolyzed. Unsubstituted  polynuclear aromatic hydrocarbons (PAH) can
 be broken  down by oxidative metabolism and subsequent conjugation with polar
 molecules. The presence  of electron-withdrawing substituents tends to stabilize
 an organic molecule. Chlorines, for example, are bulky, highly electronegative
 atoms that tend to protect the nucleus of an organic molecule against chemical
 attack. Chlorinated  organic compounds tend to bioaccumulate to high levels
 because they  are easily taken  up by organisms, and, once  in the body, they
 cannot be  readily broken  down and eliminated.

 Stereochemistry
 The spatial configuration  (i.e., stereochemistry) of a neutral molecule affects its
 tendency to bioaccumulate. Molecules that are planar tend  to be more lipid-
 soluble (lipophilic) than do globular molecules of similar molecular weight. For
 neutral organic molecules, planarity can correlate with higher bioaccumulation
 unless the  molecule  is easily metabolized by  an organism.
                                                           DRAFT

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	52	

4.3            Tier I Conclusions

After consideration of all available information, one of the following conclusions is reached (Figure 1):

       •       Existing information does not provide a sufficient basis for making factual determinations.
               In this case, further evaluation in  higher tiers is appropriate.

       •       Existing information provides a sufficient basis for making factual determinations. In this
               case, one of the following decisions  is reached (Figure 1):

               •       The material  meets the exclusion criteria.

               •       The material  does not meet the exclusion criteria but information concerning the
                       potential impact of the material is sufficient to make factual determinations.
                                                                                  DRAFT

-------
53
                       DRAFT

-------
                                     54
                   EVALUATE EXISTING
                 INFORMATION; (POSSIBLE
                    LIMITED TESTING
                    FOR EXCLUSIONS)
                                        BENTHOS
    MEASUREAND
CALCULATE THEORETICAL
   BK>A<5CUMUiAtiafcl
  |>OTENTIAI; COMPARE
  MEASURE TOXICITY;
  MODEL SUSPENDED
  PHASE; DETERMINE
TOXICITY AFTER MIXING
   MEASURE TOXICITY;
       MEASURE
   BIOACCUMULATION;
 COMPARE TO FDA LIMITS
   AND TO REFERENCE
      CONDUCT
   CASE-SPECIFIC
   TOXICITY TESTS
       CONDUCT
    CASE-SPECIFIC
       TOXICITY;
   BIOACCUMULATION;
     OTHER TESTS
                                  TIER I
                           (GENERALLY REPRESENTS
                            EXISTING INFORMATION)
                                                                   TIER II
    TIER III
(GENERIC BIOASSAY
  [TOXICITY AND
BIOACCUMULATION]
     TESTS)
    TIER IV
(SPECIFIC BIOASSAY
  [TOXICITY AND
BIOACCUMULATION]
AND OTHER TESTS)
                                                                     DRAFT

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                                  55
5.0        TIER II EVALUATION

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56

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	55	

5.0           TIER II EVALUATION

Tier II provides useful information through screening tools, but not all possible determinations can be
reached at this tier. It consists of evaluation of State water quality standard (WQS) compliance using a
numerical mixing model of the disposal site conditions (Figure 2 and Appendix C) and an evaluation of
the potential for benthic impact using calculations of theoretical bioaccumulation potential (TBP) (Figure
3 and Section 10.2).

Tier II is ultimately expected to provide a reliable, rapid screen to determine potential dredged material
contaminant effects. The dredged material discharge must meet applicable WQS for all contaminants of
concern outside the mixing zone. Water column impact must also be evaluated by toxicity testing in Tier
III (Figure 2) when there  are contaminants of concern for which applicable WQS are not available or
where interactive effects are of concern.

When national sediment quality criteria (SQC) are proposed and finalized they are expected to provide
a basis for  State sediment quality standards (SQS). State SQS will be incorporated into Tier II benthic
impact evaluations. The incorporation of these standards into Tier II will be implemented in this testing
manual and regional manuals as  appropriate.

At present, only the bioaccumulation impact of nonpolar organic compounds in dredged material on
benthic organisms can be evaluated in Tier II (Figure 3). The approved procedure calculates the TBP for
a test organism  by factoring the concentration of the nonpolar organic chemical(s), the total organic
carbon in the sediment, and the percent lipid concentration in the organism. This calculation predicts the
magnitude of bioaccumulation likely to be associated with nonpolar organic contaminants in the dredged
material. Additional guidance for identifying potential bioaccumulating contaminants is provided by EPA
(1994a).
5.1           Water Column Impact

Program experience (primarily in marine, near coastal and estuarine waters) has shown that in most cases
the existing data are sufficient to make water column determinations. However, Tier I evaluation may
show that the existing information is insufficient to make a determination. If a WQS determination cannot
be made in Tier I, Tier II evaluation is mandatory (Figure 2). The discharge of dredged material cannot
cause the WQS to be exceeded outside the mixing  zone unless the State provides a variance to the
standard or waives its right to certify.
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	56	

There are two approaches for the Tier II water column evaluation for WQS compliance. One approach
is to use the numerical models provided in Appendix C as a screen, assuming that all of the contaminants
in the dredged material are released into  the water  column during the disposal  process.  The other
approach applies the same model with results from chemical analysis of the elutriate test.
5.1.1          Screen Relative To WQS

The assumption that all of the contaminants in the dredged material are completely released into the water
column during the discharge operation  is conservative because,  in virtually  all cases, most of the
contaminants remain within the dredged material. If the numerical model (Appendix C) predicts that the
concentrations of all contaminants of concern after consideration of mixing are less than the available,
applicable WQS, the dredged material complies with WQS. If the screen/model, as applied indicates that
the WQS is exceeded, the elutriate analysis approach (Section S.I.2) should be employed.
5.1.2          Elutriate Analysis Relative To WQS

For an elutriate analysis, the numerical mixing model (Appendix C) is run with chemical data obtained
from an elutriate test conducted on the dredged material. The standard elutriate analysis is described in
Section 10.1.2.1 and the analytical procedures for measuring constituents in the water are provided in
Section 9.4.2. The  model  is, in effect,  using  data  that more accurately represent the contaminant
concentrations that will be present in the water column after consideration of mixing. If the numerical
model (Appendix  C) predicts that the concentration of all  contaminants  of concern at the edge of the
mixing zone is  less than the available, applicable WQS,  the dredged material complies with WQS.
Otherwise, it does not.
5.2            Benthic Impact

The currently available Tier II procedure for evaluating potential benthic impact consists of evaluating
the TBP, calculated according to the guidance in Section 10.2. A comparison is made between the TBP
calculated  for  the nonpolar organic contaminants of concern  in dredged material  and for the same
constituents in the reference sediment. At present, this calculation can be performed for nonpolar organic
compounds,  but not for polar organic compounds,  organometals, or metals. If  such  constituents are
contaminants of concern in a dredged material requiring bioaccumulation evaluation, further evaluation
has to take place in Tier III.
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	57	

Even if the dredged material contains other contaminants of concern than nonpolar organic contaminants,
it is still useful to calculate the TBP. The TBP provides an indication of the magnitude of bioaccumulation
of nonpolar organics that may be encountered in actual testing (Tiers III and/or IV). Additionally, the
calculation may eliminate the need for further evaluation of nonpolar organics and thereby reduce efforts
in higher tiers.
5.3           Tier II Conclusions

One of two possible conclusions is reached regarding the potential water column impact of the proposed
dredged material:

       •      The available WQS requirements are met. Further information on water column toxicity
              must be evaluated in Tier III when there are contaminants of concern for which applicable
              WQS are not available or where interactive effects are of concern.

       •      Concentrations of one or more of the dissolved contaminants of concern, after allowance
              for mixing, exceed available WQS beyond the boundaries of the mixing zone. In this
              case, the proposed discharge of dredged material does not comply with WQS.

For nonpolar organics, one of the following conclusions is reached based on comparison between the TBP
for the dredged material and for the same contaminants in the reference sediment:

       •      The TBP for the nonpolar organic contaminants of concern in the dredged material does
              not exceed the TBP for the reference sediment and,  therefore, the dredged material is
              predicted not to result in benthic  bioaccumulation of the measured non-polar organic
              compounds. However, further evaluation of biological effects in Tier III is required.

       •      The TBP for the  nonpolar organic contaminants of concern  in the dredged material
              exceeds the TBP for the reference sediment. In this case, the information is not sufficient
              to predict whether the dredged material will result in benthic bioaccumulation of the
              measured non-polar organic compounds, and further evaluation of bioaccumulation in Tier
              III is required.
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                                        58
                   EVALUATE EXISTING
                 INFORMATION; (POSSIBLE
                    LIMITED TESTING
                   FOR EXCLUSIONS)
    WATER COLUMN
      BENTHOS
    MEASURE AND
  MODEL DISSOLVED
   CONTAMINANTS;
  COMPARE TO WQS
CALCULATE THEORETICAL
   BIOACCUMULATION
  POTENTIAL; COMPARE
    TO REFERENCE
  MQOei SUSPENDED
•  PWSg;PSTER**NS
TOXICmr AFTER *4JXJKG
  MEASURE TOXiCITY?
      MEASURE
  etOAOOUMULATtON;
COMPARE TO FDA UMTFS
  AMD TO REFERENCE
      CONDUCT
   CASE-SPECIFIC
   TOXICITY TESTS
      CONDUCT
    CASE-SPECIFIC
      TOXICITY;
  BIOACCUMULATION;
     OTHER TESTS
                                 TIER I
                          (GENERALLY REPRESENTS
                           EXISTING INFORMATION)
     TIER II
(SOLELY CONCERNED
 WITH CHEMISTRY)
                                                                  TIER 10
                                                              fGENERlC-BtQASSAY
                                                                   TESTS)
     TIER IV
 (SPECIFIC BIOASSAY
   tTOXICITY AND
 BIOACCUMULATION]
 AND OTHER TESTS)
                                                                        DRAFT

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	59	

6.0           TIER HI EVALUATION

Tier III testing assesses the impact of contaminants in the dredged material on appropriately sensitive and
benchmark  organisms  to determine  if  there  is  the  potential  for  an unacceptable  (toxicity  or
bioaccumulation) impact at the disposal site. Lists of candidate test species (Sections 11 and 12: Tables
11-13) include consideration of: (1) appropriate sensitivity such that testing cannot occur with insensitive
organisms; (2) allowing appropriate Regional flexibility based on the list provided in this manual or the
approved regional implementation manual; (3) providing some benchmark species for comparing (where
appropriate) the sensitivity of regional  species not widely used for such testing.

The Tier HI assessment methods are bioassays (toxicity and bioaccumulation tests) (Figures 1 through 3).
Generic guidance provided in this manual may have to be modified for specific species. Where possible
and appropriate, organisms representative of the water column and benthic biota  and conditions at the
disposal site or the appropriate reference area should be used. Also, exposure routes must be appropriate
(e.g., benthic test species  must be truly benthic, that is, living on or in the sediment).

Presently, Tier  III  toxicity  tests primarily  use lethality as the endpoint. Chronic/sublethal tests for
sediments are under development; none are considered to be currently suitable for wide-spread national
use and hence are not included in this manual. New,  appropriate benthic and water column tests,
including  sediment chronic/sublethal  tests, will  be included  in future revisions  of this  manual as
appropriate.

The recommended procedures for water-column toxicity tests (Figure 2) use appropriate sensitive water
column organisms (Section 11.1.1, Table 11). The assay for benthic impact (Figure 3) uses deposited
sediment and appropriately sensitive benthic organisms (Section 11.2.1, Table 12).

Bioaccumulation also has  to  be  considered to fully evaluate potential benthic impact (Figure 3). The
results of bioaccumulation  tests are  used  to predict  the  potential  for uptake of  dredged-material
contaminants by organisms (Kay, 1984).

Tier III information is usually sufficient for making factual determinations. Only in unusual cases  is
further information on toxicity or bioaccumulation (or both) required at Tier IV.
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	60	

6.1            Water Column Toxicity Tests

Tier III (Figure 2) considers the effects on water  column organisms,  after allowance for  mixing, of
dissolved contaminants plus those associated with suspended participates. The toxicity and mixing data
results are generated as described in Section 11.1.

After considering the tests and considering mixing, one of the following conclusions is reached:
               If the 100% dredged material elutriate toxicity is not statistically higher than the dilution
               water (see Section 8.0, Table 3), the dredged material is not predicted to be acutely toxic
               to water column organisms.

               The concentration of dissolved plus suspended contaminants, after allowance for mixing,
               does not exceed 0.01 of the toxic (LCX or ECjo) concentration beyond the boundaries of
               the mixing zone. Therefore the dredged material is predicted not to be acutely toxic to
               water column organisms.  However,  benthic  impact has to be  considered.  If the
               information warrants, it is acceptable to determine water column effects at Tier III and
               benthic effects at another tier.

               The concentration of dissolved plus suspended contaminants, after allowance for mixing,
               exceeds 0.01 of the toxic (LCX or ECX) concentration beyond the boundaries of the mix-
               ing zone. Therefore, the dredged material is predicted to be acutely toxic to water column
               organisms.
6.2            Benthic Toxicity Tests

Evaluation of benthic (i.e., sediment) toxicity tests in Tier III (Figure 3) is based on data generated
according to the guidance in Section  11.2. Dredged material is predicted to be acutely toxic to benthic
organisms when mean test organism mortality:

        •       is statistically greater  than in the reference sediment, and

        •       exceeds mortality (or  other appropriate end point) in the reference sediment by at least
               10% (the 10% value should be used unless  a different value has been developed for
               specific test species and end-points for regulatory use, and is technically defensible; e.g.,
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	61	

              a 20% value for lethality can be used for the amphipods Ampelisca abdita, Rhepoxynius
              abronius and Eohaustorius estuarius (Swartz et al., 1985; Mearns et al.,  1986;  SAIC,
              1992a,b)).

However, even if there is a certain level of toxicity (e.g., marginal mortalities for a single non-benchmark
species), the preponderance of evidence could suggest that the sediment  is not acutely toxic to benthic
organisms. Acute toxicity testing of contaminants in the dredged material  in Tier III will result in one of
the following possible conclusions:

       •      Mortality (or other appropriate endpoint) in the dredged material is not statistically greater
              than in the reference sediment,  or does not exceed  mortality (or  other appropriate
              endpoint) in the reference sediment  by at least 10%. Therefore, the dredged material is
              predicted not to be acutely toxic to benthic  organisms.  However, bioaccumulation of
              contaminants also has to be considered. If the information warrants,  it is acceptable to
              determine benthic toxicity at Tier III and bioaccumulation at another tier.

       •      Mortality (or other appropriate endpoint) in the dredged material is statistically greater
              than in the reference  sediment and exceeds mortality (or  other appropriate endpoint) in
              the reference sediment by at least 10%. In this case, the dredged material is predicted to
              be acutely toxic to benthic organisms.
6.3           Benthic Bioaccumulation

Body burdens of chemicals are of concern for both ecological and human health reasons. The Tier III
benthic bioaccumulation tests (Section 12.1) are conducted for a subset of the contaminant of concern list
based on the contaminant bioaccumulation properties discussed in Sections 4.2 and 10.2. These tests
provide for the determination of  bioavailability  through 28-day  exposure tests. For  purposes of
comparison with an action or tolerance level such as from Food and Drug Administration (FDA) as
described below (or when conducting a Tier IV risk assessment), the duration of a bioaccumulation test
should be sufficient for organisms to reach steady-state tissue residues for all compounds. However, the
time to reach or approach steady-state varies among different compounds and, to a lesser extent, among
species. Test designs that assure that steady-state has been attained require a large number of samples and
substantial  expense. As a cost-effective compromise, it is recommended that a 28 day exposure be used
for the "standard"  bedded sediment bioaccumulation test for neutral organics and metals. For most
compounds, 80% to 100% of steady-state tissue residues will be attained within 28 days. For a few
slowly accumulated compounds, such as dioxin, DDT compounds, and PCBs, 28 days is inadequate to
                                                                                DRAFT

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	62	

attain 80% of steady-state (Boese and Lee, 1992). For these compounds, steady-state tissue residues
should be estimated by multiplying the 28-day tissue residue values by a "steady-state correction factor".
The "steady-state correction factor" is simply the reciprocal of the decimal  fraction of the amount of
steady-state tissue residue obtained after 28 days. For example, if the tissue residue after 28 days is 0.33
of the residue at 90 days (or whatever duration is required to obtain steady state), the correction factor
would be 1/0.33 =  3. These correction values should be obtained from previously conducted lab studies
(e.g., Boese and Lee, 1992). Data evaluated to date indicate that the correction factor is on the order of
2 to 4 (e.g., see Lee et al., 1994). Thus, in the absence of specific laboratory studies a correction factor
of 3 is recommended.

Bioaccumulation of most compounds, if it occurs, will be detectable after the 28-day exposure period,
even though steady state may not have been reached. Thus, Tier III bioaccumulation tests provide useful
information about the potential for bioaccumulation (i.e., bioavailability), even when steady-state tissue
residues are not determined, e.g.  when comparing to a reference sediment.

Concentrations of contaminants of concern in tissues of benthic organisms following dredged material
exposure are compared to applicable Food and Drug Administration (FDA) Action or Tolerance Levels
for Poisonous or Deleterious Substances in Fish and Shellfish for Human Food, when such levels (i.e.,
limits) have been set for the contaminants. The FDA levels (Table 2) are based on human-health as well
as economic  considerations (21 CFR  109 and 509), but do  not indicate the potential for environmental
impact on the contaminated organisms or the potential for biomagnification. Because contamination of
food in  excess of FDA levels is considered  a threat to human health, EPA and USAGE consider
concentrations in excess of such levels in any test species to be predictive of benthic bioaccumulation of
contaminants. This  guidance applies even though the test species may not be  a typical human food item
partly because certain  contaminants can be transferred through aquatic food  webs, but  mainly because
uptake to FDA levels in relatively short term tests with  one species may indicate the potential for
accumulation in other species.

Based on tissue comparisons with FDA levels, one of the following conclusions is reached:

        •      Tissue concentrations of one or more contaminants  are not statistically less than the FDA
               levels.  Therefore, the  dredged material  is predicted to result in benthic bioaccumulation
               of contaminants.

        •      Tissue  concentrations  of all contaminants either are statistically less than FDA levels or
               there are no FDA levels for the contaminants. In this case, the information is insufficient
               to reach a conclusion with respect to benthic bioaccumulation of contaminants. The
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	63	

Table 2.       Food and Drug Administration (FDA) Action Levels for Poisonous and Deleterious
              Substances in Fish and Shellfish for Human Food."
Substance                                                                   Action Level1*

Metals

  Methyl Mercury                                                            1.0 ppm

Pesticides

  Benzene Hexachloride (BHC)                                                 0.3 ppm
  Chlordane                                                                  0.3 ppm
  Chlordecone (Kepone)                                                       0.3 ppm
  DDT + DDE                                                              5.0 ppm
  Dichlorophenoxyacetic acid                                                  1.0 ppm
  Dieldrin + Aldrin                                                          0.3 ppm
  Endrin                                                                    0.3 ppm
  Fluridone                                                                  0.5 ppm
  Heptachlor + Heptachlor Epoxide                                            0.3 ppm
  Hexachlorobenzene (HBC)                                                   0.3 ppm
  Isopropylamine                                                             0.25 ppm
  Mirex                                                                     0.1 ppm
  Simazine                                                                   12.0 ppm
  Toxaphene                                                                 5.0 ppm

Industrial Chemicals

  PCBsc                                                                     (2.0 ppm)
    Action levels are established, revised, and revoked through notices published in the Federal Register.
    It is the responsibility of the users of the list to keep up to date on any amendments to this list. For
    further information on current action levels, users may contact the Food and Drug Administration,
    Center for Food Safety and Applied Nutrition, Industry Programs Branch [HFF-326, 200 C Street,
    S.W., Washington, DC 10204; (202) 205-5251].

    Action levels are reported in wet weight.

    There is no FDA action level for PCBs as a tolerance level has now been established (21 CFR part
    109.30), which is equal to die previous action level.
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	64	

       dredged material has to be further evaluated in Tier III as described below for bioaccumulation
       potential before a conclusion can be reached.

Tissue contaminant concentrations following exposure to dredged material which are statistically less than
FDA levels, or for which there are no such levels, are compared to tissue contaminant concentrations for
organisms similarly exposed to reference sediment. One of the following conclusions is reached based
on this comparison:

     •  Tissue concentrations of contaminants of concern in organisms exposed to dredged material do
       not statistically exceed those of organisms exposed to the reference sediment; therefore, the
       dredged material is predicted not to result in benthic bioaccumulation of contaminants. However,
       benthic toxicity effects also have to be considered.

     •  Tissue concentrations  of contaminants of concern in organisms  exposed to  dredged material
       statistically exceed those of  organisms exposed to the reference material. In this case, the  final
       conclusion regarding benthic bioaccumulation of contaminants would be based upon technical
       evaluations that emphasize the various factors deemed appropriate in a particular region (see last
       paragraph in this section). Additional testing (Tier IV) may be required.

One other possibility exists: tissue concentrations are above FDA limits but are not statistically different
from the reference (or disposal) site. This situation represents an exceptional case which can only be dealt
with at the regional level.

The above comparisons to FDA values address human health concerns, and follow from EPA/USACE
(1991). Other approaches which  should be considered in addition to the use of FDA values include
comparisons to state fish advisories,  cancer and non-cancer risk models,  existing ambient  fish
concentration data. State fish advisories  exist for the following chemicals for which EPA risk-based
screening values are  being developed:  (carcinogens) chlordane,  DDT,  dieldrin,  hexachlorobenzene,
lindane,  toxaphene,   PAH,  PCBs,  2,3,7,8-TCDD; (noncarcinogens)  endosulfan, mirex, cadmium,
mercury, selenium, endrin. Methods to  calculate carcinogenic and non-carcinogenic health risks are
summarized in EPA (1989a).  "Computerized Risk and Bioaccumulation  System", an  expert system for
PC computers, is available to predict tissue residues in sediment-dwelling  shellfish and the associated
excess cancer risk (Lee et  al., 1990). Note that this program does not  calculate risks  associated  with
mobile invertebrates or fishes, and  that it should be used only to supplement data derived from other
methods.
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	65	

Reference comparisons are made  for the protection of aquatic  life as well as human health because
bioaccumulation is both undesirable and an indicator of bioavailability (Figure 3). It is recognized that
residue effects information does not exist to fully interpret bioaccumulation data; the approach followed
in this manual is the best presently available.

When the bioaccumulation of contaminants in dredged-material tests statistically exceeds that in reference-
material tests, five factors should be assessed. Where available,  regional  guidance should be consulted
regarding the relative importance of these factors:

    •  What is the lexicological importance of the contaminants (e.g., Do they biomagnify? Do they
       have effects at low concentrations?) whose bioaccumulation from the dredged material statistically
       exceeds that from the reference material?

    •  By what magnitude does bioaccumulation from the dredged material exceed bioaccumulation from
       the reference material?

    •  What is the  propensity for the contaminants  with statistically significant bioaccumulation to
       biomagnify within aquatic food webs (Kay, 1984)? Contaminants which biomagnify appear to be
       few in number but widespread, and include DDT, PCB,  methylmercury and, possibly, dioxins
       and furans.

    •  What is the magnitude by which contaminants whose bioaccumulation from the dredged material
       exceeds that  from  the reference material also  exceeds the concentrations found in comparable
       species living in the vicinity of the proposed disposal site?

    •  For how many contaminants is bioaccumulation from the dredged material  statistically greater than
       bioaccumulation from the reference material?
6.4    Tier III Conclusions

The above five factors and perhaps other factors are complexly interrelated; i.e., the importance of each
factor depends on its interaction with all other factors. These factors have to be considered in case-
specific determinations (if needed) for dredged material assessed for bioaccumulation in the final step of
Tier III. After considering these factors, one of the following Tier III conclusions is reached:
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	66	

 • Discharge of the dredged material is predicted not to result in above-reference toxicity or benthic
   bioaccumulation of contaminants.

 • Discharge of  the  dredged  material  is  predicted  to result  in above-reference  toxicity or
   bioaccumulation of contaminants.

 • Further information is required to make factual determinations, specifically in Tier IV.
                                                                              DRAFT

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67
                       DRAFT

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                                   68
                   EVALUATE EXISTING
                INFORMATION; (POSSIBLE
                    LIMITED TESTING
                   FOR EXCLUSIONS)
    WATER COLUMN
      BENTHOS
    MEASURE AND
  MODEL DISSOLVED
   CONTAMINANTS;
  COMPARE TO WQS
CALCULATE THEORETICAL
   BIOACCUMULATION
 POTENTIAL; COMPARE
    TO REFERENCE
  MEASURE TOXICITY;
  MODEL SUSPENDED
  PHASE; DETERMINE
TOXICITY AFTER MIXING
  MEASURE TOXICITY;
      MEASURE
  BIOACCUMULATION;
COMPARE TO FDA LIMITS
  AND TO REFERENCE
                                       CONDUCT
                                       $e-speciR
                                       TOXK3TY;
                                       CCU*«JLATI
                                      OTHER teSTS
                                 TIER I
                          (GENERALLY REPRESENTS
                           EXISTING INFORMATION)
     TIER II
(SOLELY CONCERNED
 WITH CHEMISTRY)
     TIER III
(GENERIC BIOASSAY
   [TOXICITY AND
 BIOACCUMULATION]
      TESTS)
                                 TIER IV
                            {SPECim BIOASSAY
                             8IOACCUMULATIONJ
                             AND OTHER TESTS}
                                                                    DRAFT

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	69	

7.0            TIER IV EVALUATION

Tier IV involves case-specific, state-of-the-art testing for toxicity and/or bioaccumulation and is to be used
on a case-by-case basis only when lower tiered testing is judged to be insufficient to make complete
factual determinations. Insufficient information for a determination may include: inability to reach a clear
conclusion based on existing  data;  statistical differences are  inconclusive; evidence  is conflicting.
Experience to date suggests that Tier IV should only be used in a very few cases. When methods are
suitable for wide-spread national use, sediment chronic/sublethal testing will be part of Tier III. Until
such time as sediment chronic/sublethal tests are approved for national use in Tier III, they should only
be used in Tier IV. However, regional testing manuals may apply appropriate sediment chronic/sublethal
tests in Tier III  in advance of their inclusion in  this national manual provided this is done with a
benchmark species (Section 11.2.1) or in addition to the testing presently required in Tier III.

Tier IV tests may be  conducted for water column evaluations (Figure 2) or benthic evaluations (Figure
3). In both cases, tests should be carefully selected to address the specific issues relevant to the case in
question. Tier IV can further consider human and ecological health concerns, including risk assessment.
Case-specific evaluative criteria for Tier IV tests must be:

       •       agreed upon by EPA and USAGE and, where appropriate, the State

       •       adequate to make factual determinations.
7.1            Toxicity Tests

Tier IV toxicity tests (Figure 2) should measure end-points of clear ecological importance, for example
survival, growth and reproduction. Differences from Tier III tests may include:

       •       longer duration of exposure
       •       different species
       •       different end-points
       •       exposure in the disposal site environs.

Toxicity determinations in this tier can involve laboratory or field testing or field assessments of resident
benthic communities. Field  assessments can be difficult to interpret but can yield valuable  information
on responses of resident organisms to in-place contaminants at the dredging site as compared to a disposal
site or site environs as appropriate.
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	70	

Toxicity identification evaluation procedures (e.g., Ankley et al., 1992a) can also be used in this tier.
Such procedures can be applied to sediments when ammonia or hydrogen sulfide could be responsible
for toxicity.
7.2           Benthic Bioaccumulation

Tier IV bioaccumulation tests (Figure 3) differ from Tier III tests in that steady-state tissue concentrations
of contaminants of concern are always determined.  Such determinations  can be made by:  longer
laboratory exposures than used hi Tier III, collecting tissue samples from the field (Section 12.2.2), or
in situ exposures using transplanted organisms.

Tissue concentrations  determined in Tier IV are subject to the same comparisons  as  in  Tier  III,
specifically to FDA action limits, and to comparisons  with organisms  exposed to reference  sediment.
Conclusions possible from such comparisons and evaluative factors which should be assessed are detailed
in Section 6.3 and can include risk assessments and no effects levels for aquatic life, rather than solely
the first two comparisons.

Prediction of the movement of contaminants from sediment into and  through  pelagic food webs is
technically challenging and should only be dealt with if a Tier IV evaluation is necessary. One approach
is bioenergetic-based toxicokinetic modeling. These models have been successfully applied  to marine
(Connolly and Tonelli, 1985) and freshwater (Norstrom et al., 1976) fishes, theoretical  food chains
(Thomann, 1989), and more recently to sediment organisms (Boese et al., 1990). These models are very
data intensive to apply on a chemical and site-specific basis. It is  possible to use values determined
through QSAR (EPA,  1994a), though the default values may substantially overestimate tissue residues
in metabolizable compounds, such as PAH. Another general approach is to bracket likely concentrations
of specific contaminants at different trophic levels based on an empirical model derived from a variety
of marine food webs (Young, 1988).
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              71
PART III - SAMPLING AND ANALYSIS
                                     DRAFT

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	72	

8.0           SAMPLING

When testing is necessary, samples of dredged material, reference sediment, control sediment, organisms,
and water will be needed for physical evaluations, chemical analysis, and for bioassay tests. This section
provides general guidance for the development of a sampling plan including collection, handling and
storage.

Sampling is the foundation upon which all testing rests but there are so many case-specific factors that
influence sampling needs that detailed guidance of National scope  is impractical. Some regions of the
country have developed specific technical requirements and agency review/approvals of sampling and
analysis plans. Regional guidance from local EPA and USAGE offices should be sought for developing
project-specific sampling plans as for information gathered at Tier  I. The type of samples that may be
required to complete the evaluations of Tiers II, III, and IV are outlined in Table 3. This manual provides
general guidance on items of major importance to consider when designing a  sampling plan. Additional
guidance is provided by EPA (1994b).
8.1           Preparation For Sampling

A well-designed sampling plan is essential when evaluating the potential impact of dredged material
discharge upon the aquatic environment. Before any sampling is initiated, the sampling plan has to be
tailored to meet clearly defined  objectives  for  individual  dredging operations. Factors such as the
availability and content of historical data, the degree of sediment heterogeneity, the dredging depth, the
number  and geographical  distribution of  sample-collection   sites, the  procedures  for  collection,
preservation, storage, and tracking of samples,  and the necessity for adequate quality assurance and
quality control (Appendix G; EPA, 1994b) must be careftilly considered. The magnitude of the dredging
operation and its time and budgetary constraints should also be considered.

It is recommended that a written plan for sediment sampling and analyses be prepared  and provided to
the appropriate Federal and State agencies for coordination prior to sampling, where practicable. The Tier
I evaluation would be a logical attachment to the sampling and analysis plan for agency review and
comment. This coordination can reduce the chance of having to repeat costly procedures and can assist
in keeping projects on schedule. An adequate amount of sediment and water should be collected to
conduct planned evaluations and allow for any contingencies. Maximum allowable and recommended
sample and organism holding times as well as the exigencies of resampling should be given careful
consideration.
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The importance of sampling is underscored by the fact that any evaluation is only as complete and reliable
as the sampling (and sample handling and storage) upon which it is based. Thus, inadequacies or biases
in sampling will limit the accuracy and/or the usefulness of the study results.

The primary objective of sediment and water collection is to obtain samples to adequately and accurately
characterize the dredging and reference area. Sample size should be large enough to attain the appropriate
detection limits but small enough to be conveniently handled and transported within the requirements for
all planned analyses. The quality of the information obtained through the testing process  is impacted by
the following four factors:

       •      collecting representative samples
       •      collecting an appropriate number of samples
       •      using appropriate sampling techniques
       •      protecting or preserving the samples until they  are tested.

Ideally, the importance  of each of these three factors will  be fully  understood and appropriately
implemented.  In practice, however,  this is not always the case. There may be occasions when study
needs, time, costs or other resource constraints will limit the amount of information that should or can
be gathered. When this is the case, the relative importance of each of these factors has  to be carefully
considered in light of the specific study purposes.

An important  component of any field sampling program is a preproject meeting with all concerned
personnel.  Personnel involved  may  include management, field personnel, laboratory personnel, data
management/analysis personnel, and representatives of regulatory agencies, the permit applicant, and the
dredging company. To assure sampling quality, at least one individual familiar with the study area should
be included in the preproject meeting. The purposes of the meeting include:

       •      defining the objectives of the sampling program
       •      ensuring communication among participating groups
       •      ensuring agreement on methods, QA/QC details and contingency plans.

The more explicitly the objectives of a testing program can be stated, the easier it  will be to design an
appropriate sampling plan. A complete sampling plan will result in a level of detail such that all sampling
procedures and locations are clearly defined,  sample volumes are clearly established, all  logistical
concerns are fully addressed, and target analytes are identified to class of compound.
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	75	

8.2           Components Of A Sampling Plan

The following steps will help to ensure that all essential sampling plan information is provided:

       •      Review the plan for the proposed dredging operation, including the dimensions of the
              dredging area, the dredging depth(s), side-slopes, the volume of sediment for disposal,
              and the type of dredge equipment (e.g., clamshell, hydraulic) for determining  composite
              sampling or delineating representative project segments.
       •      Evaluate the prior history and the existing database for the area, in particular, information
              gathered in Tier I. Identify relevant data and the need for additional data. Identify areas
              of potential environmental concern within the confines of the dredging operation.
       •      If appropriate, subdivide the dredging  area into project segments on the basis of an
              assessment of level of environmental concern within the dredging area. This may be an
              iterative process that starts before sampling,  using available  information, and that is
              refined after sampling, based on new data.
       •      Determine the number of samples to be collected and select sampling locations. Choose
              methods and equipment for positioning vessels at established stations.
       •      Determine what sampling methods will be used.
       •      Define procedures for sample handling, preservation, storage, and  (if applicable) field or
              shipboard analysis.
       •      Identify logistical considerations and safety precautions.

The subsections that follow discuss each of these steps and provide general guidance for their conduct.
An essential step, preparation of a quality assurance/quality control (QA/QC) project plan, is discussed
in detail in Appendix G and EPA (1994b)  and  must be integral to the project.  The QA/QC plan is
essential to ensure that there will be sufficient and appropriate data of known and documented quality to
make decisions with confidence  and to defend  those decisions. Properly prepared, a  QA/QC  plan
expedites project coordination.
8.2.1         Review of Dredging Plan

A review of the plan for the dredging operation provides a basis for determining the sampling strategy.
The volume of material  to be dredged and the method of dredging are two important factors which will
help to determine the number of samples required. The  number of samples required  is generally a
judgement which considers the cost, resolution, and the risk of an incorrect decision regarding the volume
of material to be dredged.  Knowledge of the depth and physical characteristics of the  material to be
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	76	
dredged will help to determine the kind of sampling equipment that is required. The boundaries of the
dredging area have to be known to ensure that the number and location of samples are appropriate.
Sampling should generally be to the project depth (including overdredging) unless the sediments are
known to be vertically homogeneous.
8.2.2          Historical Data

All information relevant to the dredging site should be reviewed. Using pertinent available information
to determine project segments and station locations within the dredging area is both cost and technically
effective. If a review of historical data identifies possible sources of contamination, skewing the sampling
effort toward these areas may be justified for thorough characterization of these areas, but can lead to an
incomplete  assessment of contamination in  the whole area.  In  areas  of  unequally distributed
contamination, the total sampling effort should be increased to ensure representative, but not necessarily
equal, sampling of the entire site.  Sediment sampling techniques are detailed in Mudroch and MacKnight
(1991). The information gathered for the Tier I evaluation (discussed in Section 4.1) should be reviewed
for assistance  in designing the sampling plan, in particular the following:

       •      Geotechnical and hydrodynamic data
               The grain size, specific gravity, water or solids content, total organic carbon (TOC) and
               identification of sediment horizons are helpful in making operational decisions. Areas of
               high currents and high wave energy tend to have larger grain-sized sediments than do
               quieter areas. Many contaminants have a greater affinity for clay and silt than for sand.
               Horizontal  and vertical gradients may exist  within the sediment. Local  groundwater
               quality and movement should be determined if groundwater  is a potential source of
               contamination.

       •      Quality and age of available data
               The value of the available data should be critically weighed. Existing high-quality data
               might lower costs by reducing the number of analytes measured or tests required for the
               proposed dredging operation. Existing data that do not meet all  quality assurance/quality
               control  (QA/QC)  standards  may still  be useful  if appropriate  calibration  and
               documentation are available; they are less useful if older methods with higher detection
               limits  were used. Information from such studies might be helpful in identifying areas of
               contamination, but not  in accurately assessing the degree of contamination.
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       •      Known distribution of contaminants
              All evidence regarding contaminants within or near the dredging area, including spill
              data, may be an important consideration in  identifying locations for sampling and/or
              determining sampling intensity.

       •      Dredging history
              Knowledge of prior dredging may  dramatically  affect sampling plans.  If the area is
              frequently dredged (every  1-2 years) or if the sediments are subject to frequent mixing
              by wave  action, currents,  or ship  traffic, the sediments are likely  to  be relatively
              homogeneous.  Assuming that there is no major contaminant  input, the sampling effort
              may be minimal. However, if there  is  information regarding possible contamination or
              heterogeneity is possible,  a more extensive sampling  effort may be indicated. New
              excavations of material  unaffected by anthropogenic input may  require less  intensive
              sampling than maintenance dredging.
8.2.3         Subdivision of Dredging Area

Sediment characteristics are likely to vary substantially within the limits of the area to be dredged as a
result of geographical and hydrological features. Areas of low hydrodynamic energy will be characterized
by fine sediments that have  a  greater tendency to accumulate contaminants than do coarser-grained
sediments.  (However note that contaminants, if present  in  coarse-grained  sediments,  may  be more
bioavailable than if present in fine-grained sediments). Sediments in and downstream of heavily urbanized
or industrialized areas are more likely to accumulate contaminants than sediments farther removed from
direct contaminant input.

Many dredging operations can be subdivided into project segments (horizontal and/or vertical) which can
be treated as separate management units.  A project segment is an area expected to  have relatively
consistent characteristics that differ substantially from the characteristics of adjacent segments. Project
segments may be sampled with various intensities and, if warranted by  the study objectives and test
results, the dredged material from various project segments can be managed differently during dredging
and disposal to limit environmental impact. When the sampling plan is developed, project segments can
be designated, based on factors including but not limited to: historical data, sediment characteristics,
geographical  configuration,  anticipated method of dredging, depth of  cut, sampling- or dredging-
equipment limitations, results of pilot studies, and  known or  suspected contaminant  concentrations.
Surface sediments might be considered separately from subsurface sediments at the same location if
vertical stratification of contamination  is expected or encountered. Large dredging operations located
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	78	

within industrialized areas might require subdivision into several project segments horizontally and into
one or more segments vertically.  A dredging operation characterized by relatively uniform distribution
of sediment type in a nonindustdalized location might be considered as a single project segment. Vertical
subdivisions usually are not appropriate in areas of rapid shoaling or in areas of high sediment mixing
by ship scour, which are likely to be relatively homogenous vertically. Vertical subdivisions smaller than
about 1 m are usually impractical because dredge operators generally cannot reliably control excavation
with any finer precision; vertical subdivisions should reflect the actual removal precision to be employed
during the dredging operation. If analytical data and test results for two or more project segments prove
to be similar,  these segments may be treated as one larger segment when considering disposal options.
If the analytical and test results demonstrate important differences between project segments, alternative
disposal options may be necessary for portions of the  total sediment volume.

Any established sampling program should be sufficiently flexible  to  allow changes based  on field
observations;  however, any deviations  from the sampling plan must be documented, along  with the
rationale for such  deviations. Certain characteristics of the sediments, such as color  or texture, can be
an indication of patchiness. The greater the patchiness, the larger the number  of samples that will be
required to  adequately characterize the area. The project manager can refine a sampling program based
on historical data and/or a preliminary sampling survey of the dredging area.
8.2.4          Selection of Sampling Locations and Number of Samples

Generally a single sampling strategy will be adequate for most circumstances. However, in some cases,
two sampling strategies may be required. For instance, when sampling involves both uncontaminated and
highly contaminated sediments with interfaces between the two,  a single sampling strategy may not be
sufficient to adequately characterize these sediments, which will  probably be treated differently.

The method of dredging, the volume of sediment to be removed, the areal extent of the dredging project,
and the horizontal and vertical heterogeneity of the sediment are key to determining station locations and
the number of samples to be collected  for the total  dredging  operation and for  each project segment.
When appropriate to testing objectives,  samples  may be composited prior to analysis (with attention to
the discussion later in this section). The appropriate number of samples and the proper use of compositing
should be  determined for each operation on a case-by-case  basis. Note  that the following detailed
discussion  is not appropriate to all dredging operations. Sampling a number of small, isolated shoals is
very different than sampling a large, contiguous open area.
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	79	

Factors to Consider:

The following factors, many of which follow from information gathered in Tier I, should be among those
considered in sampling station and pattern selection:

       •      objectives of the testing program
       •      bathymetry
       •      area of the dredging project
       •      accessibility
       •      flows (currents, tides)
       •      mixing (hydrology)
       •      sediment heterogeneity
       •      contaminant source locations
       •      land use activities
       •      available resources
       •      other physical characteristics.

Station Locations:

Station locations within the dredging area should include locations downstream from major point sources
and in quiescent areas,  such as turning basins, side channels, and inside channel bends, where fine-
grained sediments are most likely to settle. Characteristics which help to define the representativeness of
station(s) within a segment include:

       •      The distribution of sediments to be dredged is clearly defined.
       •      The project segment being sampled is clearly defined.
       •      The sampling locations are distributed appropriately within each project segment.
       •      Multiple samples should be collected if sample variability is suspected.
       •      When sediment variability is unknown, it may be necessary  to conduct a preliminary
              survey of the dredging area to better define the final sampling program.

Sample Replication:

Within a station, samples  may be collected for replicate testing. For this manual, laboratory replicates
are generally recommended as opposed to field replicates. The former (subsamples of a composite sample
of the replicates) involves pseudo-replication compared to separate samples for each replicate, but is more
appropriate for dredged material evaluations where sediments will be homogenized by the dredging and
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discharge process. The latter involves true replication but is more appropriate for field investigations of
the extent and degree (or not) of homogeneity of sediment toxicity.

Depth Considerations:

Sediment composition can vary vertically as well  as horizontally. Samples should be collected over the
entire dredging depth (including over-dredging), unless the sediments are known to be vertically homoge-
neous or there are adequate data to demonstrate that contamination does not extend throughout the depth
to be excavated. Separate analyses of defined sediment horizons may be useful to determine the vertical
distribution of contamination if warranted by the study objectives. A major consideration of vertical
compositing is the anticipated depth  of dredging.  For example,  even though sediments in a 1 m shoal
may vary in composition, the material would be mixed as a result of the dredging process.

Sampling Bias:

Ideally, the composition of an area and the composition of the samples obtained from that area will be
the same. However, in practice, there often are differences due to bias in the sampling program, including
disproportionate intensity of sampling in different parts of the dredging area and equipment limitations.

In some cases, to minimize bias, it may be useful to develop a sampling grid for each  project segment.
The horizontal dimensions of each project segment may be subdivided into grid cells of equal size, which
are numbered sequentially. Cells are then selected for sampling either randomly or in an stratified random
manner. It can be important to  collect more than the minimum number of samples required, especially
in areas suspected of having high or highly variable contamination.  In some cases, although additional
costs and logistic considerations will  apply, extra samples may be archived (for long time periods in the
case of physical characterization or chemical analyses and for short time periods in the case of biological
tests), should reexamination of particular project segment(s) be warranted.

In other cases, a sampling grid may not be desirable. This is particularly the case where dredging sites
are not continuous open areas, but are rather a series of separate humps, bumps, reaches and pockets with
varying depths and surface  areas. In these latter cases, sample  distribution is  commonly biased with
intent.

Level of Effort:

In some cases, it may be advisable to consider varying the level of sampling effort. Project segments
suspected or known to be contaminated  may be targeted for an increased  level of effort so that the
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boundaries and characteristics of the contamination  can be identified. A weighting approach can be
applied whereby project segments are ranked in increasing order of concern, and level of concern can
then be used as a factor when determining the number of samples within each project segment relative
to other project segments.

Number of Samples:

In general, the number of samples that should be collected within each  project segment is inversely
proportional to the amount of known information, and is proportional to the level of confidence that is
desired in the results and the suspected level of contamination. No specific guidance can be provided, but
the following factors should be considered:

       •      the greater the number of samples collected, the better the areal and/or vertical definition

       •      single measurements are inadequate to describe variability

       •      the means of several measurements at each station within a project segment generally are
              less variable than individual measurements at each  station.

Time and Funding Constraints:

In all cases, the ultimate objective is to obtain sufficient information to evaluate the environmental impact
of a dredged  material  disposal operation. The realities of time  and  funding constraints have to  be
recognized, although such do not justify inadequate environmental evaluation. Possible responses to cost
constraints have been discussed by Higgins (1988). If the original sampling design does not seem to fit
time or funding constraints, several options are available, all of which increase the risk of an incorrect
determination:

       •      Reduce the number of project segments into which the project is divided, but maintain the
              same total number of samples.

       •      Maintain (or even increase)  the number of stations sampled, and composite  multiple
              samples from within a project segment so that a lower number of analyses are performed
              per project segment.
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Project Segments:

Regardless of the final decision on project segments and the number of sample stations and replicates per
project segment, expected or known degree of contamination will  be the dominant factor in initially
describing the proposed project segments. If variation in potential dredged material impact within a
project segment is likely, where possible it may be advisable either to use a stratified random-sampling
approach or to redefine project-segment boundaries. Once sampling  data are  available, it is advisable to
reconsider the boundaries of the project segments to be used  in the actual dredging in order to maximize
homogeneity within segments.

Sample Compositing:

The objective of obtaining an accurate representation and definition of the dredging area and method has
to be  satisfied  when compositing samples.  Compositing provides a  way to control  cost while still
analyzing sediments from a large number of stations. Compositing results in a less detailed description
of the area sampled than would individual analysis at each station. However if, for example, five analyses
can be performed  to characterize a project segment,  the increased  coverage afforded by collecting 15
individual samples and combining sets of three into five composite samples for analysis may justify the
increased time and cost of collecting the extra 10 samples. Compositing can also provide the large sample
volumes  required for some biological  tests.  Composite  samples represent the  "average"  of the
characteristics of the individual samples  making up  the composite and  are generally appropriate for
logistic and other reasons; however, composite samples which serve to "dilute" a highly toxic but
localized  sediment "hot spot"  are not recommended. Further, compositing should generally  avoid
combining physical/chemical samples or stations with very different sediment grain size characteristics.

Sample Definition:

When  a sediment sample is collected,  a decision has to be made as to whether  the entire sediment volume
is to be considered as the sample or whether the sediment volume represents separate samples.  For
instance, based on observed stratification, the top 1 m of a core might be considered to be  a separate
sample from the remainder of the core. After the sediment to be considered as a sample is identified, it
should be thoroughly homogenized.  Samples may be split before  compositing,  with one half of the
original sediment archived for possible later analysis, and the other half combined with parts of other
samples. These are  then thoroughly homogenized (using clean instruments until  color  and textural
homogeneity are achieved), producing the composite sample.
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8.2.5         Sample Collection Methods
                                           i
Sample collection requires an adequately trained crew, an adequate vessel equipped with navigational and
supporting equipment appropriate to the site and the study, and noncontaminating sampling apparatus
capable of obtaining representative samples. Divers may  also be used in some cases to collect some
samples; in such cases divers must be certified and approved diver safety management plans must be in
place. To assure sampling quality, at least one individual familiar with the study area should be present
during the sampling activities. Sampling effort for a proposed dredging operation  is primarily oriented
toward collection of sediment samples for physical and chemical characterization and for biological tests.
Collection of water samples is  also required to evaluate potential water column impact. Collection of
organisms near  the disposal site might be  necessary if  there  is a need  to  characterize  indigenous
populations or to assess  concentrations of contaminants in tissues.  Organisms for use in toxicity  and
bioaccumulation tests may also  be field-collected.

In general, a hierarchy for sample collection should be established  to prevent contamination from the
previous sample, especially when using the same sampling apparatus  to collect samples for different
analyses. Where possible, the known, or expected, least contaminated stations should be sampled first.
At a station where water and sediment are to be collected, water samples should  be collected prior to
sediment samples. The vessel should ideally be positioned downwind or  downcurrent of the sampling
device. When raising or lowering sampling devices, care should be taken to avoid  visible surface slicks
and the vessel's exhaust. The deck and sample handling area should be kept clean to help reduce the
possibility of contamination.
8.2.5.1                       Sediment Sample Collection

Mudroch and MacKnight (1991) provide useful reference information. Higgins and Lee (1987) provide
a perspective on sediment collection and analysis as commonly practiced in USAGE Districts. ASTM
(1991a) and Burton (1991) provide guidelines for collecting sediments for lexicological testing. Guidance
provided in these publications may be followed on all points that do not conflict with this manual.

Care should be taken to avoid contamination  of sediment samples during  collection and handling. A
detailed procedure for handling sampling equipment and sample containers should be clearly stated in the
sampling plan associated with a specific project. This may be accomplished by using standard operating
procedures (SOPs). For example, samples designated for trace metal analysis should not come into contact
with metal surfaces (except stainless steel, unless specifically prohibited for a project), and samples
designated for organic analysis should not come into contact with plastic surfaces. Samples for biological
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tests may be stored in clean polypropylene containers. Subsamples for particular groups of analytes may
be removed from areas of the sample not in physical contact with the collecting instrument.

A coring device with appropriate liners is recommended whenever sampling to depth is required. The
choice of corer design depends upon factors including the objectives of the sampling program, sediment
volumes required for testing, sediment type, water depth, sediment depth, and currents or tides. A gravity
corer may be limited to cores of 1-2 m in depth, depending upon sediment grain size, degree of sediment
compactness, and velocity  of the drop. For penetration greater than 2 m, a vibratory corer or a piston
corer is generally preferable. These types of coring devices are generally limited to soft, unconsolidated
sediments. A split-spoon core may be used for more compacted sediment. The length of core that can be
collected is usually limited to 10 core diameters in sand substrate and 20 core diameters in clay substrate.
Longer cores can be obtained, but substantial sample disturbance results from internal friction between
the sample and the core liner.

Freefall  cores can cause compaction of the vertical structure of sediment samples. Therefore, if the
vertical stratification in a core sample is of interest, a piston or vibra corer should be used. Piston corers
use both gravity and hydrostatic pressure. As the cutting edge penetrates the sediments, an internal piston
remains  at the level of the  sediment/water interface, preventing sediment compression  and overcoming
internal friction. A vibra corer is a more complex piece of equipment but is  capable of obtaining 3- to
7-m cores in a wide range of sediment types by vibrating a large diameter core barrel through the
sediment column with little compaction. If the samples will not be sectioned prior to analysis, compaction
is not a problem, and noncontaminating freefall corers are a suitable alternative.

Corers are the samplers of preference in most cases because of the variation in contamination with depth
that can occur in sediment deposits. Substantial variation with depth is less likely in shallow channel areas
without major direct contaminant inputs, that have frequent ship traffic, and  from which sediments are
dredged at short intervals.  Generally, in these situations,  accumulating sediments are resuspended and
mixed semicontinuously by ship scour and turbulence,  effectively preventing stratification. In such cases,
surface grab samples can be representative of the mixed sediment column, and  corers should be necessary
only if excavation of infrequently disturbed sediments below the mixed layer is planned.

Grab samplers are also appropriate for collecting surficial samples of reference or control sediments. A
grab can be Teflon-coated to prevent potential contamination of trace metal samples. The sampling device
should at least be rinsed  with clean water between samples and possibly also solvent-rinsed.
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	85	

8.2.5.2                      Water Sample Collection

If water samples are necessary, they should be collected with either a noncontaminating pump or a dis-
crete water sampler. When sampling with a pump, the potential for contamination can be minimized by
using a peristaltic or a magnetically coupled impeller-design pump. The system should be flushed with
the equivalent of 10 times the volume of the collection tubing. Also, any components within several
meters of the sample intake should be noncontaminating (i.e., sheathed in polypropylene or epoxy-
coated). Potential sample contamination must be avoided, including vessel emissions and other sampling
apparatus.

A discrete water sampler should be of the close/open/close type so that only the target water sample
comes into contact with internal sampler  surfaces.  Seals should be Teflon-coated whenever possible.
Water sampling devices should be acid-rinsed (1:1 nitric acid) prior to use for collection of trace-metal
samples, and solvent-rinsed prior to collection of samples for organic analyses.
8.2.5.3                      Organism Collection

Benthic organism collection methods may be species specific and can include, but are not restricted to,
bottom trawling, grabs or  cores. If organisms are to  be maintained  alive, they should be transferred
immediately to containers with clean, well-oxygenated water, and sediment as appropriate. Care must be
taken to prevent organisms from coming  into contact with potentially contaminated areas or fuels, oils,
natural rubber, trace metals, or other contaminants.
8.2.6         Sample Handling, Preservation, and Storage

Detailed procedures for sample handling, preservation, and storage should be part of the  standard
operating procedures and protocols developed for each sampling operation. Samples are subject to
chemical, biological, and physical changes as soon as they are collected. Sample handling, preservation,
and storage techniques have to be designed to minimize any changes in composition of the sample by
retarding chemical and/or biological  activity and by avoiding contamination.  Collection methods, volume
requirements, container specifications, preservation techniques, storage conditions and holding times
(from the time of sample collection) for sediment, water, and tissue samples are discussed below and
summarized in Table 4.
8.2.6.1                      Sample Handling

Sufficient sample volume must be collected to:


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	91	

        •       perform the necessary analyses

        •       partition  the samples,  either in the field or as soon  as possible after  sampling,  for
               respective storage and/or analytical requirements (e.g., freezing for trace metal analysis,
               refrigeration for bioassays)

        •       archive portions of the sample for possible later analysis.

Sample handling is project and analysis specific as well as being based on what is practical and possible.
Generally, samples to be analyzed for trace metals should not come into contact with metals, and samples
to be analyzed for organic compounds should not come into contact with plastics. All sample containers
should be appropriately cleaned (acid-rinsed for analysis of metals; solvent-rinsed for analysis of organic
compounds).

For analysis of volatile compounds, samples should completely fill the storage container, leaving no air-
space. These samples should be refrigerated but never frozen or the containers will crack. Samples  for
other kinds of chemical analysis are sometimes frozen. If the sample is to be frozen, just enough air space
should be allowed for expansion to take place. Container labels have to withstand soaking, drying, and
freezing without becoming detached or illegible. The labelling system should be tested prior to use in  the
field.

Sediment  samples for biological testing should have at least the larger living organisms removed from
the sediment prior to testing. This may be accomplished by press-sieving the sediments through a 1-mm-
mesh screen. Other matter retained on the screen with the organisms,  such as shell fragments, gravel,
and debris, should be recorded and discarded. Prior to use in bioassays, individual test sediments should
be thoroughly homogenized with clean instruments  (until color  and textural homogeneity is achieved).
8.2.6.2                       Sample Preservation

Preservation steps  should be  taken  immediately upon sediment  collection.  There  is no  universal
preservation or storage technique although storage in the dark at 4°C is generally used for all samples
held for any length of time prior to partitioning, and for some samples after partitioning. A technique for
one group of analyses  may interfere with other analyses. This problem can be overcome by collecting
sufficient sample volume to utilize specific preservation or storage techniques for specific analytes or
tests. Preservation, whether by refrigeration, freezing, or addition of chemicals, should be accomplished
                                                                                 DRAFT

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	92	

onboard the collecting vessel whenever possible. If final preservation techniques cannot be implemented
in the field, the sample should be temporarily preserved in a manner that retains its integrity.

Onboard refrigeration is generally accomplished with coolers and ice; however, samples  should be
segregated from melting ice or cooling water. Samples which are to be frozen on board may be stored
in an onboard freezer or may simply be placed in a cooler with dry ice or blue ice. Sediment samples
for biological analysis should be preserved at 4°C, never frozen or dried. Additional guidance on sample
preservation is given in Table 4.
8.2.6.3                       Sample Storage

The elapsed time between sample collection and analysis should be as short as possible. Sample holding
times for chemical evaluations  are analysis-specific (Table 4). Sediments for bioassay (toxicity and/or
bioaccumulation) testing should be tested as soon as possible, preferably within 2 weeks of collection.
Studies to  date suggest that sediment storage time should  not exceed 8 weeks (at 4°C, in the dark,
excluding air) (Becker and Ginn, 1990; Tatem et al., 1991). Toxicity may change with storage time.
Sample storage conditions (e.g., temperature, location of samples) should be documented.
8.2.7         Logistical Considerations and Safety Precautions

A number of frustrations in sample collection and handling can be minimized  by carefully thinking
through the process and requirements before going to the field (e.g., see EPA, 1994b). Contingency plans
are essential. Well trained, qualified, and experienced field crews should be used. Backup equipment and
sampling gear, and appropriate repair parts, are advisable. A surplus of sampling containers and field data
sheets should be available. Sufficient ice and adequate ice-chest capacity should be provided, and the
necessity of replenishing ice before reaching the laboratory should be considered. A vessel with adequate
deck  space is safer  and allows for more efficient work than an overcrowded vessel. Unforeseeable
circumstances (e.g., weather delays) are to be expected in field sampling, and time to  adequately deal
with the unforeseen has to be included in sampling schedules.

Appropriate safety and health precautions must be observed during field sampling activities. EPA (1984)
should be used as a guidance document to prepare a site-specific health  and safety plan. The health and
safety plan should be prepared as a separate document from the QA project plan. Requirements set forth
in the Occupational Safety and Health Administration 29 CFR § 1910.120 (Federal Register, Vol. 54,
No. 43) should be met for medical surveillance, personal protection,  respirator fit testing (if applicable),
and hazardous waste operations training (if applicable) by all personnel working in contaminated areas
or working with contaminated media.
                                                                                 DRAFT

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	93	

The procedures and practices established in the site-specific health and safety plan must be observed by
all individuals participating in the field activities. Safety requirements should also be met by all observers
present during field audits and inspections. The plan should include the following information:

       •      site location and history

       •      scope of work

       •      site control

       •      hazard assessment (chemical and physical hazards)

       •      levels of protection and required safety equipment

       •      field monitoring requirements

       •      decontamination

       •      training and medical monitoring requirements

       •      emergency planning and emergency contacts.

Samples must be properly disposed when  no longer needed (e.g., see EPA,  1994b). Ordinary sample-
disposal methods are usually acceptable, and special precautions are seldom appropriate. Under Federal
law [40 CFR 261.5(a)], where highly contaminated wastes are involved, if the waste generated is less
than 100 Kg per month, the generator is  conditionally exempt  as a small-quantity generator and may
accumulate up to 1,000 Kg of waste on the property without being subject to the requirements of Federal
hazardous waste regulations. However, State  and local regulations may require special handling and
disposal of contaminated samples.  When samples  have to be shipped, 49 CFR  100-177 should be
consulted for current Department of Transportation regulations on packing and shipping.


8.2.8         Non-Indigenous Test Species

Over the last few years, there has been a growing awareness of the ecological and economic damage
caused by introduced species. Because both east and west coast species are often used in bioaccumulation
tests, there is a real potential of introducing bioaccumulation test species or associated fauna and flora
(e.g., pathogens, algae used in transporting the worms). It is the responsibility of the persons conducting
the bioaccumulation or toxicity tests to assure that no non-indigenous species are  released.
                                                                                DRAFT

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	94	

The general procedures to contain non-indigenous species are to collect  and then poison all water,
sediment, organisms and associated packing materials (e.g., algae, sediment) before disposal. Chlorine
bleach can be used as the poison. A double containment system is used to keep any spillage from going
down the drain. Guidance on procedures used in toxicity tests can be found  in Appendix B of DeWitt et
al.  (1992a). Flow-through tests can generate large quantities of water, and researchers should plan on
having sufficient storage facilities.
                                                                                 DRAFT

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	95	

9.0           PHYSICAL ANALYSIS OF  SEDIMENT  AND CHEMICAL  ANALYSIS OF
              SEDIMENT, WATER, AND TISSUE SAMPLES

This section provides guidance on the selection of chemical and physical analyses to aid in the evaluation
of dredged material for proposed disposal, and on the methods used to analyze these parameters. QA/QC
guidance is provided in Appendix G and EPA (1994b).

The methods cited in this section may be used to develop the required chemical information. However,
other methods may provide similar results, and the final choice of analytical procedures depends upon
the needs of each evaluation. In all cases, proven,  state-of-the-art methods should be used.

Any dredged material from estuarine or marine areas contains salt. The salt can interfere with the results
obtained from some analytical methods. Any methods proposed for the analysis of sediment and water
from estuarine or marine environments must explicitly address steps taken to control salt interference.
9.1           Physical Analysis of Sediment

Physical characteristics of the dredged material must be determined to help assess the impact of disposal
on the benthic environment and the water column at the disposal site. This is the first step in the overall
process  of sediment  characterization, and also  helps to identify  appropriate control and  reference
sediments for biological tests. In addition, physical analyses can be helpful in evaluating the results of
analyses and  tests conducted later in the characterization process.

The general analyses may include (1) grain size, (2) total solids and (3) specific gravity.

Grain-size analysis defines the frequency distribution of the size ranges of the particles that make up the
project sediment (e.g., Plumb, 1981; Folk, 1980). The general size classes of gravel, sand, silt, and clay
are the most useful in describing the size distribution of particles in dredged-material samples. Use of the
Unified Soil Classification System (USCS) for physical characterization is recommended for the purpose
of consistency with USAGE engineering evaluations (ASTM, 1992).

Total solids is a gravimetric determination of the organic and inorganic material remaining in a sample
after it has been dried at a  specified temperature. The total solids values generally are used to convert
concentrations of contaminants from a wet weight to a dry weight basis.
                                                                               DRAFT

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	96	

The specific gravity of a sample is the ratio of the mass of a given volume of material to an equal volume
of distilled water at the same temperature (Plumb, 1981). The specific gravity of a dredged-material
sample helps to predict the behavior (i.e., dispersal and settling characteristics) of dredged material after
disposal.

Other physical/engineering properties (e.g.,  Atterburg limits, hydrometer analysis, settling properties,
etc.) may be needed to evaluate the quality of any effluent discharged from confined disposal facilities.
Guidance in this regard is provided in Appendix B.
9.2           Target Detection Limits

The selection of appropriate target detection limits (TDLs) is vital (e.g., TetraTech, 1986a; EPA, 1986a).
TDLs  should be lower than the appropriate values against which the data are to be  compared  for
interpretation. Different analytical methods are capable of detecting different concentrations of a chemical
in a sample.  For example, a highly sensitive technique can detect a much lower chemical concentration
than can a screening technique for the same chemical. The accuracy of measurements also differs among
analytical techniques. In general, as the sensitivity and accuracy of a technique increases, so does the
cost.  Recommended TDLs that are judged to be feasible,  cost effective, and to meet the requirements
for dredged material evaluations are summarized in EPA (1994b),  along with example analytical methods
that are capable of  meeting  those TDLs.  However, any method  that can  achieve those TDLs is
acceptable, provided that the appropriate documentation of the method performance is generated for the
project.

The TDL is a performance goal set between the lowest, technically feasible  detection limit for routine
analytical methods and available regulatory criteria or guidelines  for evaluating dredged material.  The
TDL is, therefore, equal to or greater than the lowest amount of a chemical that can be reliably detected
based on the variability  of the blank  response of routine analytical methods (see EPA  [1994b]  for
discussion of method blank response).  However, the reliability  of a chemical measurement generally
increases as the concentration increases. Analytical costs may also be lower at higher detection limits.
For these reasons, the TDLs in EPA (1994b) have been set  at not less than 10 times lower than available
regional or international dredged  material guidelines for  potential biological effects associated with
sediment chemical contamination.

All data generated for dredged material  evaluation should meet  the  TDLs in EPA (1994b) unless
prevented by sample-specific  interferences. Any sample-specific interferences must be well  documented
by the laboratory. If significantly higher or lower TDLs  are required to meet rigorously defined data
                                                                                DRAFT

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	97	

quality objectives (e.g., for human health risk assessments) for a specific project then, on a project-
specific basis, modification to existing analytical procedures may be necessary. Such modifications must
be documented in the QA project plan.  An experienced analytical chemist should be consulted so the
most appropriate method modifications can be assessed, the appropriate coordination with the analytical
laboratory can be implemented, and the data quality objectives can be met. A more detailed discussion
of method modifications is provided in EPA (1994b).
9.3           Chemical Analysis of Sediment

9.3.1         Target Analytes

Chemical analysis provides information about the chemicals present in the dredged material that, if
biologically available, could cause toxicity and/or be bioaccumulated. This information is valuable for
exposure assessment and for deciding which of the contaminants present in the dredged material to
measure in tissue samples.

If the historical review conducted in Tier  I (Section 4.1) establishes  a reason to believe that sediment
contaminants may be present, but fails to  produce sufficient information to develop a definitive list of
potential contaminants, a list of target analytes has to be compiled. Target analytes should be selected
from, but  not necessarily limited  to, the  compounds in Table 5  and  from the historical review
information. The target list should include contaminants that historical  information or commercial and/or
agricultural applications suggest could be present at a specific dredging site — for example, tributyltin
near shipyards, berthing areas, and marinas where these compounds have been applied. Analysis of
polynuclear aromatic hydrocarbons (PAH) in dredged material should focus on those PAH compounds
that are on the priority pollutant list (Clarke and Gibson, 1987).

All PCB analyses  should be made using congener-specific methods. The sum of the concentrations of
specific congeners is an appropriate measure of total PCBs (NOAA, 1989).

Sediments should be analyzed for total organic carbon (TOC). This is  particularly important if there are
hydrophobic  organics on the contaminant of concern list developed  in Tier I.  The TOC  content of
sediment is a measure of the total amount of oxidizable organic material in  a sample and also affects
contaminant bioaccumulation by, and  effects to, organisms (e.g.,  Di  Toro et al., 1991; DeWitt et al.,
1992b).
                                                                                DRAFT

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                                                          98
Table 5.
Potential Contaminants of Concern Listed According to Structural Compound Class.
Structural Compound
Class
                            Contaminant
                                               Structural Compound
                                               Clan
                                                                                               Contaminant
Phenols
Substituted Phenols
Organonitrogen
Compounds
Low Molecular Weight
Polynuclear Aromatic
Hydrocarbons (PAH)
High Molecular Weight
Polynuclear Aromatic
Hydrocarbons (PAH)
Chlorinated Aromatic
Hydrocarbons
Chlorinate Aliphatic
Hydrocarbons
           phenol
           2,4-dimethylphenol
           2-methylphenol
           4-methylphenol

           2,4,6-trtehlorophenol
           para-chloro-meta-cresol
           2-chk>rophenol
           2,4-dichtorophenol
           2-nitro phenol
           4-nrtrophenol
           2,4-dinitrophenol
           4,6-dinitro-o-cresol
           pentachlorophenol

           benzidine
           3,3'-dichk>robenzidine
           2,4-dinitrotoluene
           2,6-dinitrotoluene
           1 ,2-diphenylhydrazine
           nitrobenzene
           /V-nftrosodimethylamine
           /V-nitrosodiphenylamine
           /V-nitrosodipropylamine

           acenaphthene
           naphthalene
           acenaphthylene
           anthracene
           phenanthrene
           fluorene
           1 -methylnapthalene
           2 -methylnapthalene

           fluoranthene
           benzofcVanthracene
           benzofr^pyrene
           benzo/bjfluoranthene
           benzole Wuoranthene
           chrysene
           dibenzofc,/>yanthracene
           idenod ,2,3-crf)pyrene
           pyrene

           1 ,2,4-trichlorobenzene
           hexachlorobenzene
           2-chloronaphthalene
           1 ,2-dichlorobenzene
           1 ,3-dichlorobenzene
           1 ,4-dichk>robenzene

           hexachlorobutadiene
           hexachloroethane
           hexachlorocyck) pentad iene
                                                                 Halogenated Ethers
                                                                 Phthalates
Polychlorinated
Biphenyls (PCB)
as Arochlors8
Miscellaneous
Oxygenated
Compounds

Pesticides
bis(2-chloroethyl)ether
4-chlorophenyl ether
4-bromophenyl ether
bis(2-chloroisopropyl)
ether
bis(2-chlorethoxy)methane

bis(2-ethylhexyl)phthalate
butyl benzyl phthalate
di-n-butyl phthalate
di-n-octyl phthalate
diethyl phthalate
dimethyl phthalate

PCB-1242
PCB-1254
PCB-1221
PCB-1232
PCB-1248
PCB-12 60
PCB-1016

TCDD (dk>xin)b
PCDF (furan)
isophorone

aldrin
dieldrin
chtordane
chlorbenside
dacthal
DDT°
endosulfand
endrin
endrin aldehyde
heptachlor
heptachlor epoxide
a-hexachtorocyctohexane
^-hexachlorocyctohexana
4-hexachk>rocyck>hexane
K-hexachlorocyclohexane
toxaphene
mirex
methoxychlor
parathion
malathion
guthion
demeton
                                                                                                    DRAFT

-------
                                                        99
Table 5.  (continued)
Structural Compound
Class
Volatile Halogenated
Alkanes
Volatile Halogenated
Alkenes
Volatile Aromatic
Hydrocarbons
Chlorinated
Benzenes
Contaminant
                                   Structural Compound
                                   Class
tetrechloromethane
1,2-dichloroethane
1,1,1 -trichloroethane
1,1-dichloroethane
1,1,2-trichloroethane
1,1,2,2-tetrachloroethane
chloroethane
chloroform
1,2-dichloropropane
dichloromethane
chloromethane
bromomethane
bromoform
dichlorobromoethane
fluorotrichloromethane
dichlorodifluoromethane
chlorodibromomethane

1,1 -dichlorethylene
1,2-f/-a/7s-dichlorethylene
trans-1,3-dichloropropene
c/s-1,3-dichloropropene
tetrachlorethene
trichlorethene
vinyl chloride

benzene
ethylbenzene
toluene

1,3-dichlorobenzene
1,4-dichlorobenzene
1,2-dichlorobenzene
1,2,4-trichlorobenzene
hexachlorobenzene
Volatile Unsaturated
Carbonyl Compounds

Volatile Ethers
                                                               Metals
                                                              Miscellaneous
                             Contaminant
acrolein
acrylonitrile

2-chlorethylvinylether
bis(chloromethyl)ether
                             aluminum
                             antimony
                             arsenic
                             beryllium
                             butyl tins
                             cadmium
                             chromium (hexavalent)
                             cobalt
                             copper
                             iron
                             lead
                             manganese
                             mercury
                             nickel
                             selenium
                             silver
                             thallium
                             tin
                             zinc

                             ammonia*
                             asbestos
                             benzoic acid
                             cyanide
                             guaiacols
                             methylethyl ketone
                             resin acids
'It is recommended that PCS analyses use congener-specific methods. The sum of the concentrations of specific congeners is
an appropriate measure of total PCBs (see Table 7).
bAdditional dioxin and furan (e.g., TCDF) compounds are listed in Table 6.
'Includes DDT, DDD, and DDE
dlncludes 
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	100	

Sediments in which metals are suspected to be contaminants of concern may also be analyzed for acid
volatile sulfide (AVS) (Di Toro et  al., 1990; EPA, 1991a). Although  acceptable guidance on the
interpretation of AVS  measurements is not yet available, and AVS  measurements  are not generally
recommended at this time, such measurements can provide information on the bioavailability of metals
in anoxic sediments. Presently,  AVS studies  represent  an area of on-going research which may be
formally included in the manual if and when decision criteria are determined.
9.3.2         Selection of Analytical Techniques

Once the list of target analytes for sediments has been established, analytical  methods have  to be
determined. The methods will, to some degree, dictate the amount of sediment sample required for each
analysis. General sample sizes are provided in Table 4, and include possible requirements for more than
one analysis for each group of analytes. The amount of sample used in an analysis affects the detection
limits attainable by a particular method.

TOC analyses should be based on high-temperature combustion rather than on chemical oxidation.  Some
classes of organic compounds are not fully degraded by chemical/ultraviolet techniques. The volatile and
nonvolatile  organic components make up the TOC  of  a sample.  Because inorganic carbon  (e.g.,
carbonates and bicarbonates) can be a significant proportion of the total carbon in some sediment, the
sample has to be treated with acid to remove  the inorganic carbon prior to TOC analysis. The method
of Plumb (1981)  recommends HC1 as the acid.  An alternative choice might be sulfuric acid since it is
nonvolatile, is used as the preservative, and does not add to the chloride burden of the sample. Whatever
acid is used, it has to be demonstrated on sodium chloride blanks that there is no interference generated
from the combined action of acid and salt in the sample. Acceptable methods for TOC analysis are
available from EPA (1994b).

For many metals  analyses in marine/estuarine  areas, the concentration of salt may  be much greater than
the analyte of interest and can cause unacceptable interferences in certain analytical techniques. In such
cases, the freshwater approach of acid digestion followed by inductively coupled plasma-atomic emission
spectrometry (ICP) or graphite furnace atomic  absorption spectroscopy (GFAAS) should be coupled with
appropriate techniques for controlling this  interference. The Hg method in EPA (1986a; Method  7471)
may be used for the analysis of Hg in sediment. Tributyltin may be analyzed by the method of Rice et
al. (1987), and selenium and arsenic by the method of EPRI (1986). A total extraction of metal ions is
neither necessary nor desirable for dredged material evaluations. The standard aqua regia extraction  yields
consistent and reproducible results.
                                                                                DRAFT

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	101	

The recommended method for analysis of semivolatile and volatile priority pollutants in sediment is
described by Tetra Tech (1986a). Analysis for organic compounds should always use capillary-column
gas chromatography (GC): gas chromatography/mass spectrometry (GC/MS) techniques for semi-volatile
and volatile  priority pollutants,  and dual  column gas  chromatography/electron-capture  detection
(GC/ECD) for pesticides and PCBs (NOAA, 1989). Alternatively, GC/MS using selected ion monitoring
can be used for PCB and pesticide analysis. These analytically sound techniques yield accurate data on
the concentrations of chemicals in the sediment matrix. The analytical techniques for semivolatile organic
compounds generally involve solvent extraction from the sediment matrix and subsequent analysis, after
cleanup, using GC or GC/MS. Extensive cleanup is necessitated by the likelihood of (1) biological macro-
molecules, (2) sulfur from sediments with low or no oxygen, and (3) oil and/or grease in the sediment.
The analysis of volatile organic compounds incorporates purge-and-trap techniques with analysis by either
GC or GC/MS. If dioxin (i.e., 2,3,7,8, - TCDD) analysis is being performed, the methods of Kuehl et
al. (1987), Smith et al. (1984), EPA (1989b; Method 8290), or EPA (1989c,d; 1990c; Method 1613) and
summary in EPA (1994b) should be consulted. EPA Method 1613 is the recommended procedure for
measuring the tetra- through octa- chlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs).
This method has been developed for analysis of water, soil, sediment, sludge, and tissue.  Table 6 shows
the 17 compounds determined by Method 1613.

Techniques for  analysis of chemical constituents have some inherent limitations for sediment samples.
Interferences  encountered  as part of the sediment matrix, particularly  in samples from heavily
contaminated  areas, may limit the ability of a method to detect or quantify some  analytes. The most
selective methods using GC/MS techniques are recommended for all nonchlorinated  organic compounds
because such analysis can often avoid problems due to matrix interferences. Gas chromatography /electron-
capture detection (GC/ECD) methods are recommended as the primary analytical tool for all PCB and
pesticide analyses because GC/ECD  analysis  will result in lower detection limits. The  analysis  and
identification  of PCBs by GC/ECD methods are based upon relative retention times  and peak shapes.
Matrix interferences may result in the reporting  of false  negatives, although congener-specific PCB
analysis reduces this concern relative to use of the historical Arochlor* matching procedure.

PCBs have traditionally been quantified with respect to Arochlor* mixtures.  This procedure can result
in errors in determining concentrations (Brown et al.,  1984).  For dredged material evaluations, the
concentration of total PCBs should be determined by summing the concentrations of specific individual
PCB congeners identified in the sample (see Table 7).  The minimum  number of PCB  congeners  that
should be analyzed are listed in the first column of Table 7 (i.e., "summation" column) (NOAA, 1989).
This summation is considered the most accurate representation of the  PCB concentration in samples.
McFarland et al. (1986) note that the most toxic PCB congeners lie mainly within the tetra-, penta-, and
hexa- chlorobiphenyl groups.  McFarland and Clarke (1989) recommend the PCB congeners in Table 7
                                                                              DRAFT

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	102	
Table 6.      PCDD and PCDF Compounds Determined by Method 1613
 Native Compound1              2,3,7,8-TCDF
                               2,3,7,8-TCDD
                             1,2,3,7,8-PeCDF
                             2,3,4,7,8-PeCDF
                             1,2,3,7,8-PeCDD
                           1,2,3,4,7,8-HxCDF
                           1,2,3,6,7,8-HxCDF
                           2,3,4,6,7,8-HxCDF
                           1,2,3,4,7,8-HxCDD
                           1,2,3,6,7,8-HxCDD
                           1,2,3,7,8,9-HxCDD
                           1,2,3,7,8,9-HxCDF
                          1,2,3,4,6,7,8-HpCDF
                         1,2,3,4,6,7,8-HpCDD
                          1,2,3,4,7,8,9-HpCDF
                                      OCDD
                                      OCDF

1 Polychlorinated dioxins and furans:
  TCDD   =  Tetrachlorodibenzo-p-dioxin
  TCDF   =  Tetrachlorodibenzofuran
  PeCDD  =  Pentachlorodibenzo-p-dioxin
  PeCDF  =  Pentachlorodibenzofuran
  HxCDD =  Hexachlorodibenzo-p-dioxin
  HxCDF  =  Hexachlorodibenzofuran
  HpCDD =  Heptachlorodibenzo-p-dioxin
  HpCDF  =  Heptachlorodibenzofuran
  OCDD  =  Octachlorodibenzo-p-dioxin
  OCDF   =  Octachlorodibenzofuran
                                                                         DRAFT

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                                           103
Table?.   Polychlorinated Biphenyl (PCB) Congeners Recommended  for Quantitation  as Potential
          Contaminants of Concern.
PCB Congener*


2,4' diCB
2,2',5 triCB
2,4,4' triCB
3,4,4' triCB
2,2',3,5' tetraCB
2,2',4,5' tetraCB
2,2',5,5' tetraCB
2,3',4,4' tetraCB
2,3',4',5 tetraCB
2,4,4',5 tetraCB
3,3',4,4' tetraCB
3,4,4' ,5 tetraCB
2,2',3,4,5' pentaCB
2,2',3,4',5 pentaCB
2,2',4,5,5' pentaCB
2,3,3',4,4' pentaCB
2,3,4,4' ,5 pentaCB
2,3',4,4',5 pentaCB
2,3',4,4',6 pentaCB
2',3,4,4',5 pentaCB
3,3',4,4',5 pentaCB
2',3,3',4,4' hexaCB
2,2',3,4,4',5' hexaCB
2,2',3,5,5',6 hexaCB
2,2',4,4',5,5' hexaCB
2,3,3',4,4',5 hexaCB
2,3,3',4,4',5 hexaCB
2,3,3',4,4',6 hexaCB
2,3',4,4',5,5' hexaCB
2,3',4,4',5',6 hexaCB
3,3',4,4',5,5' hexaCB
2,2',3,3',4,4',5 heptaCB
2,2',3,4,4',5,5' heptaCB
2,2',3,4,4',5',6 heptaCB
2,2',3,4,4',6,6' heptaCB
2,2',3,4',5,5',6 heptaCB
2,3,3',4,4',5,5' heptaCB

Congener Number1*

Summation*
8
18
28

44

52
66


77



101
105

118


126f
128
138

153





169f
170
180


187


Highest
Priority"










77

87
49
101
105

118


126f
128
138

153
156




169f
170
180
183
184



Second
Priority1

18

37
44
99
52

70
74

81




114

119
123



151


157
158
167
168





187
189

(continued)
                                                                          DRAFT

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	104

Table 7.      (continued)
PCB Congener*
2,2',3,3',4,4',5,6 octaCB
2,2',3,3',4,5,5',6' octaCB
2,2',3,3',4,4',5,5',6 nonaCB
2,2',3,3',4,4',5,5',6,6' decaCB

Congener Number1*
Highest
Summation* Priority*1
195
206
209
Second
Priority*
201
•PCB  congeners   recommended   for   quantitation,   from  dichlorobiphenyl   (diCB)   through
 decachlorobiphenyl (decaCB).

""Congeners are identified by their International Union of Pure and Applied Chemistry (IUPAC) number,
as referenced in Ballschmiter and Zell (1980) and Mullin et al. (1984).

These congeners are summed to determine total PCB concentration following the   approach  in
NOAA (1989).

dPCB congeners having highest priority for potential environmental importance based on potential for
toxicity,  frequency  of occurrence in environmental samples, and   relative  abundance in  animal
tissues (McFarland and Clarke, 1989).

ePCB congeners having second priority for potential environmental importance based on potential for
toxicity,  frequency  of  occurrence  in environmental samples, and  relative  abundance in  animal
tissues (McFarland and Clarke, 1989).

To separate PCBs 126 and  169, it is necessary to initially utilize an enrichment step with an activated
carbon column (Smith, 1981).
                                                                              DRAFT

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	105	

for analysis based  on environmental abundance,  persistence, and biological importance.  Sample
preparation for PCB congener analysis should follow the techniques described by Tetra Tech (1986a) or
EPA (1986a), but with instrumental analysis and quantification using standard capillary GC columns on
individual PCB isomers according to the methods reported by NOAA (1989) (see also Dunn et al., 1984;
Schwartz et al., 1984; Mullin et al., 1984; Stalling et al.,  1987).

Although the methods mentioned above are adequate for detecting and quantifying concentrations of those
PCB congeners comprising the majority of total PCBs hi environmental samples, they are not appropriate
for separating and quantifying PCB congeners which may coelute with other congeners and/or may be
present at relatively  small concentrations in the total PCB mixture. Included in this latter group of
compounds, for example, are PCBs 126 and 169, two of the more toxic nonortho-substituted (coplanar)
PCB congeners (Table 7). In order to separate these  (and other toxic nonortho-substituted congeners), it
is necessary to initially utilize an enrichment step with an activated carbon column (Smith, 1981). Various
types of carbon columns have been used, ranging from simple gravity columns (e.g., in a Pasteur pipette)
to more elaborate (and efficient) columns using high pressure liquid chromatography (HPLC) systems (see
Schwartz et al., 1993). The preferred method of separation and quantitation of the enriched PCB mixture
has been via high resolution GC-MS with isotope dilution (Kuehl  et al., 1991;  Ankley et al.,  1993;
Schwartz et al., 1993). However, recent studies have shown  that  if the carbon enrichment is done via
HPLC, the nonortho-substituted PCB congeners of  concern also may be  quantifiable via more widely
available GC/ECD systems (Schwartz et al., 1993).

The overall toxicity of nonortho-substituted PCBs at a site can be assessed based on a comparison with
the toxicity of 2,3,7,8-tetrachloro-dibenzo-p-dioxin (TCDD). A similar procedure can  be used for
assessing the toxicity of a mixture  of dioxins and furans. In this "toxicity equivalency factor" (TEF)
approach, potency values of individual congeners (relative to TCDD) and their respective sediment
concentrations are used to derive a "summed" 2,3,7,8-TCDD equivalent (TCDD-EQ) (EPA, 1989c; Table
8).  Ankley et al. (1992b) provide an example of the use of this approach.

TEFs have been derived for human health purposes. For aquatic organisms  the relative toxicities of
different PCB congeners and dioxins are likely to be quite  different.  For instance, wildlife or fish TEF
for PCBs are not equivalent to those for humans (Walker et al., 1992).

To  ensure that contaminants not included in the list of target analytes are not overlooked in the chemical
characterization of the dredged material,  the analytical results should  also be scrutinized by trained
personnel. The presence of  persistent major unknown analytes  should be noted.  Methods involving
GC/MS techniques for organic compounds are recommended for the identification of any unknown
analytes.
                                                                               DRAFT

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                                              106
Table 8.       Methodology for Toxicity Equivalency Factors

Because toxicity information on some dioxin and furan species is scarce, a structure-activity relationship
has been assumed.  The toxicity of each congener is expressed as a fraction of the toxicity of 2,3,7,8
TCDD.
Compound
2,3,7,8 TCDD
other TCDD
2,3,7,8-PeCDDs
other PeCDDs
2,3,7,8-HxCDDs
other HxCDDs
2,3,7,8-HpCDDs
other HpCDDs
OCDD
2,3,7,8-TCDF
other TCDFs
1,2,3,7,8-PeCDF
2,3,4,7,8-PeCDF
other PeCDFs
2,3,7,8-HxCDFs
other HxCDFs
2,3,7,8-HpCDFs
other HpCDFs
OCDF
TEF
1
0
0.5
0
0.1
0
0.01
0
0.001
0.1
0
0.05
0.5
0
0.1
0
0.01
0
0.001
                                                                               DRAFT

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	107	

9.4            Chemical Analysis of Water

9.4.1          Analytical Targets

Analysis to determine the potential release of dissolved contaminants from the dredged material (standard
elutriate) may be necessary to make a factual determination. Elutriate tests (Section 10.1.2.1) involve
mixing dredged material with dredging site water and allowing the mixture to settle. The portion of the
dredged  material that is considered to have the potential to impact the water column is the supernatant
remaining after undisturbed settling and centrifugation. Chemical analysis of the elutriate allows a direct
comparison, after allowance for mixing, to applicable water quality standards (WQS).  When collecting
samples  for elutriate testing, consideration should be given to adequate volumes of water and sediment
required to prepare samples for analysis including replicates where appropriate. In some instances, when
there is  poor settling,  the elutriate  preparation has to be  performed successively several times to
accumulate enough water for testing.

Historical water quality information from the disposal  site (Tier I) should be evaluated along with data
obtained from the chemical analysis of sediment samples to select target analytes. Chemical  evaluation
of the dredged material provides a known list of constituents which might affect the water column. All
target analytes identified in the sediment should initially be considered potential targets for water analysis.
Nonpriority-pollutant chemical components which are found in measurable concentrations in the sediments
should be included as targets if review of the literature indicates that these analytes have the potential to
bioaccumulate  in animals [i.e., have a high  K^ or  bioconcentration factor  (BCF)] and/or  are of
lexicological concern.
9.4.2          Analytical Techniques

In contrast to freshwater, there generally are no EPA approved methods for analysis of saline water
although widely accepted methods have existed for  some time  (e.g.,  Strickland and Parsons,  1972;
Grasshoff et al.,  1983; Parsons et al.,  1984). Application of the freshwater methods  to saltwater will
frequently result in higher detection limits than are common for freshwater unless care is taken to control
the effects of salt on the analytical signal. Modifications or substitute methods (e.g., additional extract
concentration steps, larger sample sizes, or concentration of extracts  to smaller volumes) might  be
necessary to properly determine analyte concentration in seawater or to meet the desired target detection
limits (TDLs). It is extremely important to ascertain a laboratory's ability to execute methods and attain
acceptable detection limits in matrices containing up to 3% sodium chloride.
                                                                                 DRAFT

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	108	

Once the list of target analytes for water has been established, analytical methods have to be determined.
The water volume required for specific analytical methods may vary. A minimum of 1  L of elutriate
should be prepared for metals analysis (as little as 100 mL may be analyzed). One liter of elutriate should
be analyzed for organic compounds. Sample size should also include the additional volume required for
the organic matrix spike and matrix spike duplicate analyses required as part of the analytical procedure.
Samples from the dredging site and, where appropriate, disposal site, should be delivered for organic and
metals analysis.  Sample  size is one of the limiting factors in determining detection limits for water
analyses, but TDLs below the WQS must be the goal hi  all  cases. Participating laboratories should
routinely report detection limits achieved for a given analyte.

Detailed methods for the analysis of organic and inorganic priority pollutants in water are referenced in
the Federal Register (1984, Vol. 49, No. 209) and in EPA (1983). Additional approved methods include
EPA (1986a,b; 1988a,b,c; 1990b,c);  APHA (1989); ASTM (1991b); Tetra Tech (1985).  Most of these
methods will require modification to achieve low detection limits  in saline waters.  Analysis of the
semivolatile organic priority pollutants involves a solvent extraction of water with an optional sample
cleanup procedure and  analysis using GC or GC/MS. The volatile priority pollutants are determined by
using purge-and-trap techniques and are analyzed by either GC or GC/MS. If dioxin (i.e., 2,3,7,8, -
TCDD) analysis is necessary, Kuehl  et al. (1987), Smith et al.  (1984), EPA (1989b;  Method 8290), or
EPA (1990c; Method 1613) should be consulted.

A primary requirement for analysis of inorganic and organic priority pollutants is to obtain detection
limits which will result in usable, quantitative  data that can subsequently be compared against applicable
WQS to determine compliance with the water quality certification requirement under Section 401. Exist-
ing EPA methods for  freshwater analysis need to be adapted  to achieve environmentally meaningful
detection limits in  saline waters  because of  matrix interferences caused by salt. For example, it is
recommended that sample extracts be concentrated to the lowest possible volume prior to instrumental
analysis, and that instrumental injection volumes be increased to lower the detection limits. All PCB and
pesticide analytes should  be analyzed by using GC/ECD, since the GC/ECD methods are more sensitive
to these compounds and will lower the detection limits. PCBs should  be quantified as specific congeners
(Mullin et al., 1984; Stalling et al., 1987) and as total  PCBs based on  the summation of particular
congeners (NOAA, 1989).

Analysis of saline water for metals is subject to matrix interferences from salts,  particularly sodium and
chloride ions, when the samples are concentrated prior to instrumental analysis.  The gold-amalgamation
method using cold-vapor atomic absorption spectrophotometry (AAS) analysis  is recommended to
eliminate saline water matrix interferences for mercury analysis. Methods using solvent extraction and
AAS analysis may be required to reduce saline water matrix interferences for other target metals. Other
                                                                                DRAFT

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	109	

methods  appropriate for metals include:  cadmium, copper, lead, iron, zinc, silver (Danielson et al.,
1978); arsenic (EPRI,  1986); selenium and antimony (Sturgeon et al., 1985); low levels of mercury
(Bloom et al., 1983); and, tributyltin  (Rice et  al.,  1987). Graphite-furnace AAS techniques  after
extraction are recommended for the analysis of metals, with the exception of mercury.
9.5           Chemical Analysis of Tissues

9.5.1         Target Analytes

Bioaccumulation is evaluated by analyzing tissues of test organisms for contaminants determined to be
of concern for a specific dredged material. Sediment contaminant data and available information on the
bioaccumulation potential of those analytes have to be interpreted to establish target compounds.

The n-octanol/water  partition coefficient  (KM)   is  used  to estimate the  BCFs of  chemicals in
organism/water systems (Chiou  et al., 1977; Kenaga  and Goring,  1980; Veith et al., 1980; Mackay,
1982). The potential for bioaccumulation generally increases as K^ increases, particularly for compounds
with log KM less than approximately 6. Above this value, there is less of a tendency for bioaccumulation
potential to increase with  increasing K^. Consequently, the relative potential for bioaccumulation of
organic compounds can be estimated from the K^ of the compounds. EPA (1985) recommends  that
compounds  for  which  the  log Kw is  greater than 3.5  be considered for further evaluation of
bioaccumulation potential. The organic compound classes of priority pollutants with the greatest potential
to bioaccumulate are PAHs, PCBs, pesticides, and some phthalate esters. Generally, the volatile organic,
phenol, and organonitrogen priority pollutants are not readily bioaccumulated, but exceptions include the
chlorinated benzenes and the chlorinated phenols. Table 9 provides data for organic priority pollutants
based on JTW. Specific target analytes for PCBs and PAHs are discussed in Section 9.3.1. The water
content and  percent  lipids  should  be routinely  determined as  part  of tissue  analyses for organic
contaminants.

Table  10 ranks the bioaccumulation potential  of the inorganic priority pollutants based on calculated
BCFs. Dredged material contaminants with BCFs greater than 1,000 Gog BCF >3) should be further
evaluated for bioaccumulation potential.

Tables 9 and  10 should be used with caution because they are based on calculated bioconcentration from
water. Sediment bioaccumulation tests, in contrast, are concerned with accumulation from a complex
medium via all possible routes of uptake. The  appropriate use of the tables is to help in selecting
                                                                                DRAFT

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	no	

Table 9.        Octanol/Water Partition Coefficients (K^) for Organic Compound Priority Pollutants and
                301(h) Pesticides.
Pollutant
Octanol/Water
Partition
Coefficients
Pollutant
Di-n-octyl phthalate              9.2
Indeno(l,2,3-crf)pyrene           7.7
Benzo(g/»0perylene              7.0
PCB-1260                       6.9
Mirex"                          6.9
Benzo(A:)fluoranthene             6.8
Benzo(6)fluoranthene             6.6
PCB-1248                       6.1
2,3,7,8-TCDD (dioxin)           6.1
Benzo(a)pyrene                  6.0
Chlordane                       6.0
PCB-1242                       6.0
4,4'-DDD                       6.0
Dibenzo(a,h)anthracene           6.0
PCB-1016                       5.9
4,4'-DDT                       5.7
4,4'-DDE                       5.7
Benzo(a)anthracene              5.6
Chrysene                        5.6
Endrin aldehyde                 5.6
Fluorantheoe                    5.5
Hexachlorocyclopentadiene       5.5
Dieldrin                        5.5
Heptachlor                      5.4
Heptachlor epoxide              5.4
Hexachlorobenzene              5.2
Di-w-butyl phthalate              5.1
4-Bromophenyl phenyl ether      5.1
Pentachlorophenol               5.0
4-Chlorophenyl phenyl ether      4.9
Pyrene                          4.9
2-Chloronaphthalene             4.7
Endrin                          4.6
PCB-1232                       4.5
Phenanthrene                    4.5
Fluorene                        4.4
Anthracene                      4.3
Methoxychlor*                  4.3
Hexachlorobutadiene             4.3
1,2,4-trichlorobenzene           4.2
Bis(2-ethylhexyl)phthalate        4.2
Octanol/Water
Partition
Coefficients
OoglU
                            Acenaphthylene                  4.1
                            Butyl benzyl phthalate            4.0
                            PCB-1221                       4.0
                            Hexachloroethane                3.9
                            Acenaphthene                   3.9
                            a-hexachlorocyclohexane         3.8
                            6-hexachlorocyclohexane         3.8
                            6-hexachlorocyclohexane         3.8
                            •y-hexachlorocyclohexane         3.8
                            Parathionb                       3.8
                            Chlorobenzene                  3.8
                            2,4,6-trichlorophenol             3.7
                            fi-endosulfan                    3.6
                            Endosulfan sulfate               3.6
                            a-endosulfan                    3.6
                            Naphthalene                     3.6
                            Fluorotrichloromethane"          3.5
                            1,4-dichlorobenzene              3.5
                            1,3-dichlorobenzene              3.4
                            1,2-dichlorobenzene              3.4
                            Toxaphene                      3.3
                            Ethylbenzene                    3.1
                            N-nitrosodiphenylamine          3.1
                            P-cbloro-m cresol                3.1
                            2,4-dichlorophenol               3.1
                            3,3'-dichlorobenzene             3.0
                            Aldrin                          3.0
                            1,2-diphenylhydrazine            2.9
                            4-nitrophenol                    2.9
                            Malathionb                      2.9
                            Tetrachloroethene                2.9
                            4,6-dinitro-o-cresol              2.8
                            Tetrachloroethene                2.6
                            Bis(2-chloroisopropyl)ether       2.6
                            1,1,1 -trichloroethane             2.5
                            Trichloroethene                  2.4
                            2,4-dimethylphenol              2.4
                            1,1,2,2-tetrachloroethane         2.4
                            Bromoform                     2.3
                            1,2-dichloropropane              2.3
                            Toluene                         2.2
                                                                                       DRAFT

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                                                  Ill
Table 9.    (continued)
Pollutant
Odanol/Water
Partition
Coefficients
Pollutant
1,1,2-trichloroethane             2.2
Guthionb                        2.2
Dichlorodiflouromethane0         2.2
2-chlorophenol                  2.2
Benzene                         2.1
Chlorodibromomethane           2.1
2,4-dinitrotoluene                2.1
2,6-dinitrotoluene                2.0
rra/«-l,2-dichloropropene        2.0
Cw-l,3-dichloropropene          2.0
Demetonb                       1.9
Chloroform                     1.9
Dichlorobromomethane           1.9
Nitrobenzene                    1.9
Benzidine                       1.8
1,1-dichloroethane               1.8
2-nitrophenol                    1.8
Isophorone                      1.7
OctanolAVater
Partition
Coefficients
                             Dimethyl phthalate              1.6
                             Chloroethane                    1.5
                             2,4-dinitrophenol                l.S
                             1,1-dichloroethylene             l.S
                             Phenol                         l.S
                             1,2-dichloroethane              1.4
                             Diethyl phthalate                1.4
                             A'-ni trosodipropy lamine          1.3
                             Dichloromethane                1.3
                             2-chloroethy Iviny lether          1.3
                             Bis(2-chloroethoxy)methane      1.3
                             Acrylonitrile                    1.2
                             Bis(2-chloroethyl)ether           1.1
                             Bromomethane                  1.0
                             Acrolein                        0.9
                             Chloromethane                  0.9
                             Vinyl chloride                  0.6
                             JV-nitrosodimethylamine          0.6
•Adapted from Tetra Tech (1985).
"•SOlfli) pesticides not on the priority pollutant list.
°No longer on priority pollutant or 301(h) list.

[Note: Mixtures, such as PCB Arochlors*, cannot have discrete K^, values, however, the value given is a rough
estimate for the mean. It is recommended that all PCB analyses use congener-specific methods. All PCB congeners
have a log K^ >4 (L. Burkhardt, EPA Duluth, pers. comm.).]
                                                                                      DRAFT

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	H2	

Table 10.   Bioconcentration Factors (BCF) of Inorganic Priority Pollutants.


Inorganic Pollutant                          Log BCF*
Metals
Methylmercury
Phenylmercury
Mercuric acetate
Copper
Zinc
Arsenic
Cadmium
Lead
Chromium IV
Chromium III
Mercury
Nickel
Thallium
Antimony
Silver
Selenium
Beryllium
Nonmetals
Cyanide
Asbestos

4.6
4.6
3.5
3.1
2.8
2.5
2.5
2.2
2.1
2.1
2.0
1.7
1.2
ND
ND
ND
ND

ND
ND
•Adapted from Tetra Tech (1986b).
"ND: No data.
                                                                              DRAFT

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	113	

contaminants of concern for bioaccumulation analysis by providing a general indication of the relative
potential for various chemicals to accumulate in tissues.

The strategy for selecting contaminants for tissue analysis should include three considerations,  all of
which are related to regulatory concern:

      •     the target analyte is a contaminant of concern and is present in the sediment as determined
            by sediment chemical analyses

      •     the target analyte has a high potential to accumulate and persist in tissues

      •     the target analyte is of toxicological concern.

Contaminants with a lower potential to bioaccumulate, but which are present at very high concentrations
in the sediments, should also be included in the target list  because bioavailability can increase with
concentration. Conversely, contaminants with a high accumulation potential and of high toxicological
concern should  be  considered as targets, even  if they are only present  at low concentrations in the
sediment. Nonpriority-pollutant contaminants which are found  in  measurable  concentrations in the
sediments should be included as targets for tissue analysis if they have the potential to bioaccumulate and
persist in tissues, and are of toxicological concern.
9.5.2       Analytical Techniques

At  present, formally approved standard methods for the analysis of priority pollutants and  other
contaminants in tissues are not available. However, studies conducted for EPA and other agencies have
developed analytical methods capable of identifying and quantifying most organic and inorganic priority
pollutants in tissues. The amount of tissue required for analysis is dependent on the analytical procedure
and the tissue moisture content. General guidance, but not firm recommendations, for the amount of
tissue required, is provided in Table 4. The required amounts may vary depending on the analytes,
matrices,  detection  limits,  and particular  analytical laboratory.  Tissue moisture content must  be
determined for each sample to convert applicable data from a wet-weight to a dry-weight basis, however
both wet- and dry-weight data should be reported.

Detection limits depend on the sample size as well as the specific analytical procedure. TDLs should be
determined for all analytes according to initial guidance in 40 CFR 136 and more definitive guidance in
                                                                                 DRAFT

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	H4	

EPA (1994b; cf. Section 9.2). Detection limits should be specified based on the intended use of the data
and specific needs of each evaluation.

Existing methods for priority pollutant tissue analysis involve two separate procedures: one for organic
compounds and another for metals. The recommended methods for the analysis of semivolatile organic
pollutants are described in NOAA (1989). The procedure involves serial extraction of homogenized tissue
samples with methylene chloride, followed by alumina and gel-permeation column cleanup procedures
that remove coextracted lipids. An automated gel-permeation procedure described by Krahn et al. (1988)
is recommended  for rapid,  efficient, reproducible sample cleanup.  The extract is concentrated  and
analyzed for semivolatile organic pollutants using GC with capillary fused-silica columns to achieve
sufficient analyte resolution. If dioxin (i.e., 2,3,7,8-TCDD) analysis is being performed, the methods of
Mehrle et al. (1988), Kuehl et al. (1987), Smith et al.  (1984), EPA (1989b; Method 8290), or EPA
(1990c; Method 1613) should be consulted.

Chlorinated hydrocarbons (e.g., PCBs and chlorinated pesticides) should be analyzed by GC/ECD. PCBs
should be quantitated as specific congeners  (Mullin et al., 1984; Stalling et al., 1987) and  not by
industrial  formulations (e.g.,  arochlors) because the  levels  of PCBs in tissues  result from complex
processes, including selective accumulation and metabolism (see the discussion of PCBs in Section 9.3.2).
Lower detection  limits and positive identification of PCBs  and pesticides can be obtained  by using
chemical ionization mass spectrometry.

The  same tissue  extract  is analyzed for other  semivolatile  pollutants (e.g.,  PAHs,  phthalate esters,
nitrosamines, phenols, etc.) using GC/MS as described by NOAA (1989), Battelle (1985), and Tetra Tech
(1986b). These GC/MS methods are similar to EPA Method 8270 for solid wastes  and soils (EPA,
1986a). Lowest detection limits are achieved by operating the mass spectrometer in the SIM mode.
Decisions to perform analysis of nonchlorinated hydrocarbons and resulting data interpretation should
consider that many of these analytes are  readily metabolized by most fish  and many  invertebrates.
Analytical methods for analysis of tissue samples for volatile priority pollutants are found in Tetra Tech
(1986b).

Tissue lipid content is of importance in  the interpretation  of bioaccumulation  information. A lipid
determination should be performed on biota submitted for organic analysis if: (1) food chain models will
be used; (2) test organisms could spawn during the test; (3) special circumstances occur (Tier  IV), such
as those requiring risk assessment. Bligh and Dyer (1959) provide an acceptable method, and the various
available methods are evaluated by Randall et al. (1991).
                                                                                DRAFT

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	115	

Analysis for priority pollutant metals involves a nitric acid or nitric acid/perchloric acid digestion of the
tissue sample and subsequent analysis of the acid extract using AAS or inductively coupled plasma (ICP)
techniques. Procedures in Tetra Tech  (1986b)  and EPA (1991c)  are generally recommended.  NOAA
(1989) methods may also be used and are recommended when very low detection levels are required.
Microwave technology may be used for tissue digestion to reduce contamination and to improve recovery
of metals (Nakashima et al., 1988). This methodology is consistent with tissue analyses performed by
NOAA (1989), except for the microwave heating steps. Mercury analysis requires the use of cold-vapor
AAS methods (EPA, 1991c). The matrix interferences encountered in analysis of metals in tissue may
require case-specific techniques for overcoming interference problems. If tributyltin analysis is being
performed, the methods of Rice et al. (1987) or Uhler et al. (1989) should be consulted.
                                                                               DRAFT

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                                       116
                   EVALUATE EXISTING
                 INFORMATION; (POSSIBLE
                    LIMITED TESTING
                    FOR EXCLUSIONS)
  MEASURE TOXICITY;
  MODEL SUSPENDED
  PHASE; DETERMINE
TOXICITY AFTER MIXING
      CONDUCT
   CASE-SPECIFIC
   TOXICITY TESTS
  MEASURE TOXICITY;
      MEASURE
  BIOACCUMULATION;
COMPARE TO FDA LIMITS
  AND TO REFERENCE
      CONDUCT
    CASE-SPECIFIC
      TOXICITY;
  BIOACCUMULATION;
    OTHER TESTS
                                 TIER I
                          (GENERALLY REPRESENTS
                           EXISTING INFORMATION)
    TIER III
(GENERIC BIOASSAY
  [TOXICITYAND
BIOACCUMULATION]
     TESTS)
    TIER IV

(SPECIFIC BIOASSAY
  [TOXICITYAND
BIOACCUMULATION]
AND OTHER TESTS)
                                                                        DRAFT

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	in	

10.0          GUIDANCE FOR PERFORMING TIER II EVALUATIONS

10.1          Ho-II:  Water Column Effects

If a water column determination cannot be made in Tier I, the Tier II water column evaluation must be
conducted for comparison with numeric water-quality standards (WQS) (Section 5.1). There are two
approaches for the Tier II water column evaluation for WQS  compliance. One approach is  to  use
numerical models provided in Appendix  C of this manual as a screen, assuming conservatively that all
of the contaminants in the dredged material are released into  the water column during the disposal
process. The other approach applies the  same model,  using the results from a chemical analysis of an
elutriate prepared from the dredged material (Section 10.1.2.1).


10.1.1        Screen Relative To WQS

A screening approach may reduce the evaluation effort for dredged material that will cause only minimal
water column impact. In a typical  disposal operation, most contaminants  remain associated with the
dredged material that settles to the bottom and cause limited water column  impact during descent. The
screen is not a requirement but is intended to reduce the effort required to develop information required
for factual determinations.

Appendix  C provides guidance on which numerical computer or analytical models should be applied to
particular  dredged material disposal projects and the information  that is necessary to perform  the
evaluations. Versions of models for use on IBM-compatible  microcomputers and example applications
are provided on the diskettes in  the pocket inside the back cover of this  manual. The output of the
appropriate model is used to determine if additional testing is needed.

The model need be run only for the contaminant of concern that requires the greatest dilution.  If this
contaminant is shown to meet the WQS, all of the other contaminants that require less dilution will also
meet the WQS. The contaminant requiring the greatest dilution is determined by calculating the dilution
that would be required to meet  the WQS. To determine the dilution D, the following equation is  solved
for each contaminant of concern in terms of dissolved concentrations:

                             D = [(C. x SS/1000) - Cy / (C^ - CJ

where        C.      =      concentration of the contaminant in the dredged material expressed as
                             micrograms per kilogram (/ig/Kg), on a dry weight basis;
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                                               118 _

               SS      =      suspended  solids  concentration  in the  dredged  material  discharge
                              expressed as grams per liter (g/L);
               1000    =      conversion factor, g to Kg;
               Q,     =      WQS in micrograms per liter (/tg/L); and
               Q.      =      background concentration of the contaminant at  the  disposal site in
                              micrograms per liter (/tg/L).
Note that if the concentration of the constituent in the dredged material (C. x SS/1000) is less than
no calculation is necessary since no dilution is required. Note also that, if the ambient disposal-site water
concentration (CJ of a constituent is greater than Q,, water quality at the disposal site cannot be met
by  dilution.  Appendix C provides  detailed information for performing the  above calculations and
identifying the contaminant of concern requiring the greatest dilution.

The concentration of this contaminant is then modeled to determine its maximum concentration in the
water column outside the boundary of the mixing zone. If this concentration is below the applicable WQS,
no additional testing is necessary to make a determination regarding WQS. If the concentration is higher,
additional testing is necessary,  as described in Section 10.1.2.

Note that the procedure described above cannot be used to evaluate water column impact. It can be used
only to determine whether additional testing for potential water-column impact, as described in Section
10.1.2, is necessary.
10.1.2         Elutriate Analysis Relative To WQS

For an elutriate analysis, the numerical mixing model (Appendix C) is run with chemical data obtained
from an elutriate test conducted on the dredged material. The standard elutriate analysis is described in
Section 10.1.2.1 and the analytical procedures for measuring constituents in the water are provided in
Section 9.4.2. The model  is, in effect, using data that more accurately represent the contaminant
concentrations that will be present in the water column after consideration of mixing. If the numerical
model (Appendix  C) predicts that the concentration of all contaminants of concern at the edge of the
mixing zone is less than the available, applicable WQS, the  dredged material complies  with WQS.
Otherwise, it does not.
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	119	

10.1.2.1                      Standard Elutriate Preparation

The standard elutriate test is used to predict the release of contaminants to the water column resulting
from open water disposal. Prior to use, all labware should be thoroughly cleaned as appropriate for the
contaminant analysis. At a minimum, labware should be washed with detergent, rinsed with acetone, five
times with tap water, placed in a clean 10% HC1 acid bath for a minimum of 4 h, rinsed five times with
tap water, and then thoroughly flushed with either distilled or deionized water.

The elutriate should be prepared by using water from the dredging site. Enough  elutriate should be
prepared for the chemical analyses and for the water column toxicity tests in Tier III.

The elutriate is prepared by subsampling approximately 1 L of the dredged material from the well-mixed
original sample.  The dredged material and unfiltered water are then combined in a sediment-to-water ratio
of 1:4 on a volume basis at room temperature (22 ±  2°C). This  is best  accomplished by volumetric
displacement.  After the correct ratio is achieved, the mixture is stirred vigorously for 30 min with a
mechanical or magnetic  stirrer. At 10 min intervals,  the  mixture is also stirred manually to ensure
complete  mixing.  After the 30 min  mixing period, the mixture is allowed to settle for 1 h.  The
supernatant is then  siphoned off without disturbing the settled material, and centrifuged to remove
particulates prior to chemical analysis (approximately 2,000 rpm  for 30 min, until visually clear). If the
elutriate is to be used for toxicity testing, refer to the procedures in Section  11.1.4.
10.1.2.2                      Chemical Analysis

Analytical procedures for specific constituents in water are provided in Section 9.4.2.


10.1.2.3                      Comparison with WQS (Standard Elutriate Test)

The model need be run only for the contaminant that requires the greatest dilution to make a WQS
determination. This contaminant may or may not be the same as that run in the screen (Section 10.1.1).
Calculations must therefore be conducted for all of the contaminants detected during analysis of the
elutriate to determine which one requires the greatest dilution. The contaminant requiring the greatest
dilution is determined by calculating the dilution that would be required to meet the WQS. To determine
the dilution D, the following equation is solved for each contaminant of concern in terms of dissolved
concentrations:
                                                                                 DRAFT

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                                              120

                             D =
       C.      =      concentration of the dissolved contaminant in the standard elutriate in micrograms
                      per liter G*g/L). All other terms are as previously defined in Section 10.1.1.
10.2           Theoretical Bioaccumulation Potential (TBP) of Nonpolar Organic Chemicals

The TBP is an approximation of the equilibrium concentration in tissues if the dredged material in
question were the only source of contaminant to the organisms. The TBP calculation in Tier II is applied
as a coarse screen to predict the magnitude of bioaccuraulation likely to be associated with nonpolar
organic contaminants in the dredged material. At present the TBP calculation can be performed only for
nonpolar organic chemicals such as PCBs. However, methods for TBP calculations with metals and polar
organic compounds are under development  and may be added to this  manual in the  future. For the
present, bioaccumulation potential of polar organic compounds, organometals, and metals in dredged
material can only  be tested (in Tiers III or IV), not calculated. However, it is still useful to calculate the
TBP,  which provides an indication of the magnitude of bioaccumulation of nonpolar organic compounds
that may be encountered in testing at higher tiers. Additionally, if the TBP of the nonpolar organic com-
pounds indicates that these contaminants are not bioavailable, this calculation may eliminate the need for
further evaluation of these compounds and thereby reduce efforts in higher tiers.

Nonpolar organic chemicals include all organic compounds that do not dissociate or form ions.  This
includes the chlorinated hydrocarbon pesticides,  many other halogenated hydrocarbons, PCBs, many
PAHs including all the priority pollutant PAHs, dioxins and furans. It does not include metals and metal
compounds, organic acids or salts, or organometallic complexes such as tributyltin or methyl mercury.

The environmental distribution of nonpolar organic chemicals is controlled largely by their solubility in
various media.  Therefore, in sediments they tend to occur primarily in association with organic matter
(Karickhoff, 1981). In organisms they are found primarily in the body fats or lipids (Konemann and van
Leeuwen, 1980;  Geyer et  al.,  1982; Mackay,  1982; Bierman, 1990).  Bioaccumulation of nonpolar
organic compounds from dredged material can be estimated from the organic carbon content of the mate-
rial, the lipid content of the organism, and the relative affinities of the chemical for sediment organic
carbon and animal lipid content.

The TBP  calculation assumes that various lipids in different organisms and organic carbon in different
sediments are similar and have similar distributional properties. Other simplifying assumptions are that
chemicals are  freely exchanged  between the  sediments   and  tissues  and that compounds  behave
                                                                                DRAFT

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	121	

conservatively. In reality, compound size and structure may influence accumulation,  and portions of
organic compounds  present  on suspended participates  may  have kinetic  or structural barriers to
availability. Another important assumption implicit in the TBP calculations is that there is no metabolic
degradation or biotransformation of the chemical. Organic-carbon normalized contaminant concentrations
are used such that the sediment-associated chemical can be characterized as totally bioavailable to the
organism. Calculations based on these assumptions yield an environmentally conservative TBP value for
the dredged material if the dredged material in question is the only source of the contaminant for the
organism. However, note that TBP calculations are not valid for sediments with TOC  ^ 0.2%.

It  is possible to  relate the concentration of a chemical  in  one phase of a two-phase system to the
concentration in the second phase when the system is in equilibrium. The TBP calculation focuses on the
equilibrium distribution of a chemical between  the dredged material or  reference sediment and the
organism. By normalizing nonpolar organic chemical concentration data for lipid content in organisms,
and organic carbon in dredged material or reference sediment, it is possible to estimate the preference
of a chemical for either phase. This approach is  based on the work of Konemann and van  Leeuwen (1980)
and Karickhoff (1981).

McFarland (1984) took the approach one step farther. He calculated that the equilibrium concentration
of nonpolar organic chemicals, which the lipids of an organism could accumulate as a result of exposure
to dredged material,  would be about 1.7  times the organic carbon-normalized concentration of the
chemical in the dredged  material. Concentrations are directly  proportional to the lipid content of the
organism and the contaminant content of the dredged material or reference sediment, and are inversely
proportional to the organic carbon content of the dredged or  reference material (Lake et al., 1987).

The possible chemical concentration in an organism's lipids [the lipid bioaccumulation potential (LBP)]
would theoretically be 1.7 times  the concentration of that chemical in the sediment  organic carbon.
Rubinstein et al. (1987) have shown, based  on  field studies with PCBs, that a value of 4 for calculating
LBP is appropriate, and this is the value that is used in this manual. However, note that in future more
precise values  for specific chemicals  may  be available.  LBP represents the potential contaminant
concentration in  lipid if the  sediment is the only source of that contaminant  to the  organism. It is
generally desirable to convert LBP to whole-body bioaccumulation potential for a particular organism of
interest. This is done by multiplying LBP by that organism's lipid content, as determined  by lipid analysis
or from reported data. Soft-bodied invertebrate lipid contents  may range from 1-2%  wet weight (based
on data from an oligochaete, midge,  and amphipod species [G. Ankley, EPA Duluth and H. Lee, EPA
Newport, pers. comm.]).
                                                                                DRAFT

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	122	

Theoretical bioaccumulation potential (TBP) can be calculated relative to the biota sediment accumulation
factor (BSAF) as
                      TBP = BSAF (C. /  %TOC) %L

where TBP is expressed on a whole-body wet-weight basis in the same units of concentration as C., and

        C,      =  concentration of nonpolar organic chemical in the dredged  material or reference
                  sediment (any units of concentration may be used);

        BSAF  =  4 (Ankley et al., 1992c)

        %TOC =  total organic carbon content of the dredged  material or reference sediment expressed
                  as a decimal fraction (i.e., 2% = 0.02); and

        %L    =  organism lipid content expressed as a decimal fraction (i.e., 3%  = 0.03) of whole-
                  body wet weight.

This calculation is based on work by McFarland and Clarke (1987), who also developed the nomograph
in Figure 4 by which TBP can be determined graphically. Using the nomograph, it is possible to quickly
estimate the TBP for organisms of various lipid contents, provided that the contaminant concentration (C.)
and organic carbon content (% TOC) of the dredged-material  or reference sediment are known. Even
though the nomograph does not provide as precise an answer as the equation, it is sufficient for Tier II
applications. Because the TBP does  not predict expected environmental concentrations but indicates the
upper range, exact evaluation is not necessary. The procedure  for using the nomograph is as follows:

        Step 1. Determine the lipid  content of an organism of interest, either from previously reported
               values or from laboratory analysis, and express the lipid content as percent of whole-body
               wet weight rather than as a  decimal fraction.

        Step 2. Locate the value on the right hand vertical axis that corresponds most  closely to that lipid
               content.

        Step 3. Follow the sloped  line until it intersects the dredged-material  or  reference-sediment
               concentration C.. C, may be expressed in any units of concentration from any of four
               ranges:  0.1-1.0; 1-10; 10-100; or 100-1,000.
                                                                                DRAFT

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                                          123
 UJ
 i—
 o
 0.

 So"

 Ss
 2 V)
 O 2
 S3
 O UJ
 mi
 1"
 o
 tr
 O
 UJ
 I
                                                                            -71  10
             SEDIMENT
         ORGANIC CARBON
                0.1
                1
                10
                100
          0.1     0.2    0.3   0.4   0.5   0.6  0.7   0.8   09   1.0
          1      2      345678g     10
          10     20    30   40    50    60   70    80    90    100
          100    200   300  400   500   600  700   800   900   1,000
                                                                      x
                                                                      3E
                                                                      V\
                                                                     C3
                                                                     o:
                                                                     o
                                                                     o
                                                                     o
                                                                     Q
                                                                     Q.
Figure 4.
                        Cs (ANY UNITS)

Nomograph for Determining Theoretical Bioaccumulation Potential. Nomograph is based
onaBSAFof4.



                                                            DRAFT

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	124	

       Step 4. From that point, read across to the left hand vertical axis and select the TBP value from
              the appropriate sediment organic  carbon column (which must be  >  0.2%  TOC)
              expressed as percent of sediment dry weight.

       Step 5. Multiply the TBP by the factor (0.1, 1, 10, 100) corresponding to the selected C,  range.
              The TBP will then be in the same units of concentration as C,.

The lipid scale and the C. scale of the nomograph can be changed by orders of magnitude by adjusting
the TBP scale in the same manner. For example, if the organism of interest is a mussel having 0.3 % lipid
content, one would simply follow the 3% lipid line  and divide the appropriate resulting theoretical
bioaccumulation  value by  10.  If the dredged material or reference sediment concentration  (C.) of a
contaminant lies  above or below the Ct ranges shown on the nomograph, the units of concentration can
be changed (e.g., change 0.02 parts per million to 20 parts per billion). Interpolation between lipid lines
or between organic carbon  columns is straightforward because all  relationships are proportional. For
example, for dredged material  or reference sediment with an organic carbon content of 3%, the TBP
would be 1/3 the TBP at 1% carbon, 5/3 the TBP at 5% organic carbon, 10/3 the TBP at 10% organic
carbon, or 20/3 the TBP at  20% organic carbon.

The following  illustration of the use of the nomograph  determines the TBP of total PCB by a fish of 6%
lipid content exposed to a sediment containing 4 ppm  PCB and 4.6% total organic carbon. Follow the
6% lipid line to a C. value of 4 and then read across to the 5% organic carbon column to obtain  a TBP
of about 19 x  1 or 19  ppm.  Because the organic carbon content of the sediment  is actually 4.6% rather
than 5%, a more precise estimate can be made by multiplying 19 by 5/4.6 to obtain a TBP of 20.6 ppm.
This value is then evaluated as detailed in Section 5.2 to determine whether a determination can be made
or further testing is necessary.
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125
                        DRAFT

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                                    126
                   EVALUATE EXISTING
                INFORMATION; (POSSIBLE
                    LIMITED TESTING
                   FOR EXCLUSIONS)
    WATER COLUMN
      BENTHOS
    MEASURE AND
  MODEL DISSOLVED
   CONTAMINANTS;
  COMPARE TO WQS
CALCULATE THEORETICAL
   BIOACCUMULATION
  POTENTIAL; COMPARE
    TO REFERENCE
  MEASURE tQXKSHYr
  MOOEU SUSPENDED
TOXICJTY AFTER MtXtNS
  ^MEASURE TOSplTY; :
       MEASURE
   BIOACCUMULATION;
 COMPARE TO FDA LIMITS
   AND TO REFERENCE
     CONDUCT
   CASE-SPECIFIC
   TOXICHY TESTS
                                       .CONDUCT  f
                                    BIOACCU MU LATION ;
                                      OTHER TESTS-  :
                                  TIER I
                          (GENERALLY REPRESENTS
                           EXISTING INFORMATION)
     TIER II
(SOLELY CONCERNED
  WITH CHEMISTRY)
     TIER III
 (GENERIC BIOASSAY
   [TOXICITY AND
 BIOACCUMULATION]
      TESTS)
                                 TIER IV
                             (SPECIFIC BIOASSAY
                               [TOXICITY AND
                             BIOACCUMULATION]
                             AND OTHER TESTS)
                                                                    DRAFT

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	127	

11.0          GUIDANCE FOR PERFORMING BIOLOGICAL EFFECTS TESTS

Biological effects tests, i.e., toxicity tests, may be necessary if Tier I evaluations conclude that the
dredged material contains contaminants which might result in an unacceptable adverse impact to the ben-
thic environment and/or the water column. Toxicity tests with whole sediment are used to determine the
potential for effects on benthic (bottom dwelling) organisms; toxicity tests with suspensions/solutions of
dredged material are conducted to determine the potential effects on water column organisms.

The objective of water column toxicity tests is to determine the potential impact  of dissolved and
suspended contaminants on organisms in the water column,  after considering mixing. Test organisms
should be representative of appropriately sensitive  water column species existing in the vicinity of the
disposal site.

The objective of benthic toxicity tests is to determine the potential impact of whole sediment on benthic
organisms at and beyond the boundaries of the disposal site. The organisms used in  testing should be
representative of appropriately sensitive infaunal or epifaunal organisms existing in the vicinity of the
disposal site. Benthic toxicity tests are intended to determine the potential chemical toxicity of a dredged
material as distinct from its physical (e.g., grain-size) effects. Some organisms are affected by differences
in sediment textures or absence of sediments (McFarland,  1981;  DeWitt et al.,  1988). Control and
reference sediments should be selected to minimize  any artifactual effects of differences in grain size. If
the sediment texture varies considerably between  the dredged material and the control or reference
sediments, any possible effects of grain size have to be determined and considered when designing the
tests and evaluating the test results (e.g., DeWitt et al., 1988).
11.1          Tier III:  Water Column Toxicity Tests

Tests to evaluate dredged-material impact on the water column involve exposing test organisms to an
elutriate dilution series containing both dissolved and suspended components of the dredged material. The
test organisms are added to the exposure chambers and exposed for a prescribed period (usually 96 h
though some tests, e.g., bivalve larvae, may be run for shorter periods).  The surviving organisms are
examined at specified intervals and/or at the end of the test to determine if the test material is producing
an effect. An introductory guide to general toxicity testing is presented in Part 8000 of APHA (1989) and
in ASTM (1991c). Biological testing aspects of these reference publications may be followed as long as
they do not conflict with  this  manual.
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11.1.1         Species Selection

Three species are recommended for use in the water column exposure and should represent different
phyla where possible (Table 11). The rationale for testing more than a single species is to cover the
potential range of differing species sensitivities and to be environmentally protective. Of the species
tested, at least one must be a sensitive benchmark (starred) species; however, this does not preclude the
use of more than one benchmark species. Those non-benchmark species listed hi Table 11 can be used
to satisfy minimum requirements if a summary of test conditions and test acceptability criteria similar to
the  starred benchmark   species  are  established,  and  data  from reference  toxicity  tests  (see
Appendix G.3.17.2) are provided on the sensitivity of the species. Species proposed for use regionally
and not listed in Table 11 must meet the species characteristics criteria, provided later hi this Section,
and proponents must generate the following supporting information:

       •      data from toxicity tests using a set of reference chemicals with differing modes of action
               demonstrating that the proposed species is as sensitive or more sensitive than the species
               in Table 11

       •      summary of test conditions and test acceptability criteria.

Note that regions cannot substitute all species in Table 11 but that, for species used regionally in addition
to the minimum requirements,  the above supporting information is desirable but not required.

The test organisms may be from healthy laboratory cultures or may be field collected, but not from within
the influence of former or active disposal sites or other discharges. Ideally, the test species should be the
same or closely related to those species that naturally dominate biological assemblages in the vicinity of
the disposal site. Species characteristics to consider when designing water-column tests include, not in
order of importance:

       •      readily available year-round
       •      tolerate handling and laboratory conditions
       •      give consistent, reproducible response to toxicants
       •      related phylogenetically and/or by ecological requirements to species characteristic of the
               water column of the disposal site area in the season of the proposed disposal
       •      standardized test protocols are available
       •      can be readily tested as juveniles or larvae to increase sensitivity
       •      important ecologically, economically, and/or recreationally
       •      appropriately sensitive.
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Table 11.     Candidate Toxicity Test Species  for Determining Potential Water Column Impact of
              Dredged Material Disposal. Details of testing procedures are provided in Appendix E.
 Crustaceans
   Mysid shrimp, Mysidopsis sp.* (N)4
                Neomysis americana* (N)
                Holmesimysis costata* (N)
   Grass shrimp,  Palaemonetes sp. (N)
   Commercial shrimp, Penaeus sp. (N)
   Cladocerans, Daphnia magna* (F)d
                Daphnia pulex* (F)d
                Ceriodaphnia dubia* (F)d

 Fish
   Silversides, Menidia sp.* (N) (E)d
   Sheepshead minnow,
 Cyprinodon variegatus* (N)d
   Speckled sanddab, Citharicthys stigmaeus (N)
   Grunion, Leuresthes tenuls (N)
   Fathead minnow,  Pimephales promelas* (F)d
 Bluegill sunfish, Lepomis macrochirus (F)
 Channel catfish, Ictalurus punctatus (F)
 Rainbow trout, Oncorhynchus mykiss* (F)

Bivalves
 Larvae of
    Oyster, Crassostreasp.* (N,E)*
    Mussel, Mytilus edulis*
Echinoderms
 Larvae of
     Sea urchins, Strongylocentrotus sp.*1*
                 (N)
                 Lytechinus pictus* (N)
     Sanddollar,  Dendrastersp.*** (N)
Note:  Examples are not presented in order of importance; however, the asterisks indicate sensitive
       recommended benchmark species. Benchmark species comprise a substantial data base, represent
       the sensitive range  of a variety of ecosystems, and provide comparative data on the relative
       sensitivity of local test species. Other species may be designated in future as benchmark species
       by EPA and USACE when the data on their response to contaminants are adequate.

*      fertilized egg to hinged, D-shaped prodissoconch I larvae. Note that these two species can be used
       in estuarine waters down to appropriate low levels of salinity (see Appendix E).
b      fertilized egg to pluteus larvae
c      sperm fertilization
d      These species can also be used in sublethal, chronic testing (methods for such testing are available
       but not detailed in this manual).

For the purpose of this manual, related to the tolerances of the test animals, (F) = Freshwater, salinity
 £ 1 %a  (N) = Near Coastal, salinity ^ 25%o   (E)  = Estuarine, salinity l-25%o. It is recognized that
the commonly accepted salinity range for estuaries is l-35%o and near coastal salinity is usually greater
than 30%c salinity.
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In addition to species occurring at the disposal site, other representative commercially available species
or sensitive life stages of economically important species may be used. Mysids of the genera Mysidopsis,
Neomysis, or Holmesimysis are highly recommended as test species. Embryo-larval stages of echinoderms,
crustaceans, molluscs, or fish are also appropriate organisms.  Adult fish  and molluscs and  large
crustaceans must not be  used for  water column toxicity testing because of their generally greater
resistance to contaminants, except as  additional test organisms where data on economically important
species are necessary to address public or regional concerns.

Regardless of theu: source, test organisms should be collected and handled as gently as possible.  They
should be gradually  acclimated to the test conditions if test conditions differ from holding conditions.
Field collected organisms must be tested within  2 weeks  of collection. Animals from established
laboratory cultures can be held indefinitely.
11.1.2         Apparatus

Water column toxicity tests are generally conducted as static exposures in pre-cleaned glass chambers
equipped with covers to minimize evaporation. The size of the chambers depends on the size of the test
species. Before use, all glassware should be washed with detergent, rinsed five times with tap water,
placed in a clean 10% HC1 acid bath for a minimum of 4 h, rinsed with acetone, five times with tap
water, and then thoroughly flushed with either distilled or deionized water.

Equipment and facilities must provide acceptable lighting requirements and temperature control. An
environmental incubator or  a  water-bath system that allows  temperature control within ±1°C  is
recommended.
11.13         Laboratory Conditions

Water column toxicity tests should be conducted under conditions known to be non-stressful to the test
organisms. Salinity for marine/estuarine organisms should be stable within ±2%x> and, for all organisms,
temperature  should  be  stable  within ±2°C  throughout  the exposure  period.  Dissolved-oxygen
concentration should not be allowed to fall below an absolute minimum of 40% saturation for warm water
species and 60% for cold water species. The temperature, salinity (if appropriate), dissolved oxygen, and
pH in the test containers should be measured and recorded daily. Measurements of other parameters, for
instance ammonia, may also be useful but need not be done daily.
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11.1.4         Laboratory Procedures

Elutriate Preparation
Elutriate should be prepared using water collected from the dredging site. Disposal site water, clean
seawater or freshwater, or artificial sea/salt mixtures should be used as  dilution water for the tests. If
sea/salt mixtures are used, they must be prepared in strict accordance with  the manufacturer's instructions
and allowed to age (with aeration) to ensure that all salts are in solution and Ph has stabilized before use
in any test.  The elutriate is prepared by subsampling approximately 1 L of the homogenized dredged-
material  sample. The dredged material  and unfiltered dredging site water  are then combined  in  a
sediment-to-water volumetric ratio of 1:4 at  room temperature (22 ± 2°C). The mixture is then stirred
vigorously for 30 min with a mechanical or magnetic stirrer. At 10 min intervals, the mixture is also
stirred manually to ensure complete mixing. After the 30 min mixing period,  the mixture is allowed to
settle for 1 h. The liquid plus the material remaining in suspension after the settling period represents the
100% liquid plus suspended paniculate phase. The supernatant is then carefully siphoned off, without
disturbing the settled material, and immediately used for testing. With some very fine-grained dredged
materials, it may be necessary to centrifuge the supernatant until the suspension is clear enough  for the
organisms to be visible in the testing chamber. Note that 15-40 L of elutriate may need to be prepared
to test some species.

Test Design
The number of replicate exposure chambers per treatment should be determined according to the guidance
in Appendix E. A minimum  of five replicates per treatment and 10 organisms  (except zooplankton or
larvae) per replicate is generally  recommended. Organism loading density must be low enough to avoid
overcrowding stress.

At least three concentrations of the dredged-material elutriate should be tested; recommended treatments
are 100%, 50%, and 10%. Water from the same source in which the animals were held prior to testing
must be  included as  a control  treatment subject  to  test survival  acceptability  criteria for controls
(Appendix G). To properly evaluate the test  results, any toxicity at 100% dilution water should also be
determined.

The test  organisms should be approximately of equal size and/or age  and assigned randomly to the
different  treatments. Zooplankton and larvae are usually transferred with the aid of a pipette. Air must
not be trapped on or under the animals during the transfer process. Larger animals may be transferred
in fine-mesh nets. Animals which are dropped or exhibit abnormal behavior should be discarded.
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The test chambers should be covered and randomly placed in an incubator or water bath. The test type
is static non-renewal; the control and test solutions are not replaced. During the exposure period, aeration
should not be supplied (unless necessary to keep dissolved oxygen concentration above 40% saturation
for warm water species or 60% for cold water species), and the test solutions should not be stirred. Some
species of crustaceans, particularly larval forms, may require feeding during the test. All food used must
be analyzed to ensure that it is free of contaminants.

Recommended test duration is 48-96 h for zooplankton and some larvae (e.g., oysters) and up to 96 h
for other organisms. For bivalve larvae, the ASTM (199Id) procedure should be used. Useful procedures
for other organisms are given in ASTM (1991c,e). For some tests, intermediate time observations may
be made of  survival but, for other tests, survival is only assessed at the end of the testing period.  For
intermediate observations, care must be taken to  minimize any stress to the test organisms. Only the
number of living organisms are counted, not the number of dead. An animal is judged dead if it does not
move either after the water is gently swirled or after a sensitive part of its body is gently touched with
a probe. At intermediate observations, a pipette or forceps is used to remove dead organisms, molted
exoskeletons, and food debris.

If greater  than acceptable mean mortality or abnormal development occurs in  the control as defined in
the procedures for proper conduct of that test, the test must be repeated. Further QA/QC considerations
are provided in Appendix G.3.17.
11.1.5         Data Presentation and Analysis

Data Presentation
The data for each test  species should be presented  in  separate tables  that include the following
information:

        •      the scientific name of the test species
        •      the number of organisms in each treatment at the start of the test
        •      the number of organisms alive at each observation period, if applicable
        •      the number of organisms recovered alive and/or in normal health from each chamber at
               the end of the test
        •      additional information including water quality and any behavioral or other abnormalities.
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Data Analysis
It is possible that no mortality or other effects will be observed in any of the treatments or that survival
or other effects in the dredged material treatments will be equal to or higher than in the control or in the
dilution water treatments. In either of these situations, there is no  need for statistical analysis and no
indication of water column toxicity attributable to the dredged material. However, if survival or other
effects in the dilution water treatment is at least 10% greater than the 100% dredged-material treatment,
the data have to be evaluated statistically to determine whether the dredged-material suspension  is
significantly more toxic than the dilution water. If the 100% dredged-material treatment is not statistically
different from the dilution water, the dredged material is predicted not to be acutely toxic to water column
organisms. An LC*, should not be calculated unless at least 50% of the test organisms die in at least one
of the serial dilutions .  If there are no mortalities greater than 50%, then the LCX is  assumed to be
^ 100%. If a statistical difference exists and greater than 50% mortality or other effects occur in all of
the treatments, it is not possible to  calculate an LCX or ECX value. If the conditions are highly toxic,
such that  the  10% treatment has  greater than 50%  mortality, further dilution must be made  (new
treatments of less than  10% dredged material) to attain a survival of greater than 50% and determine the
LCjo or ECjo by interpolation. Statistical procedures recommended for analyzing the test data are
described in detail in Appendix D.
11,1.6         Conclusions

The Tier III water-column effects evaluation involves using a numerical model comparison with the WQS.
Descriptions of the models and applications are given in Appendix C, and the models are provided on
the diskettes that can be found in the pocket inside the back cover of this manual.

The modeled concentrations of the dredged material (expressed as percentages) are compared to 0.01 of
the 48- or 96-h LCX  or  EC*,, depending on the test duration. The maximum allowable concentration
outside the mixing zone is 0.01 LCM or EC^,. Note that the 0.01 factor is intended for acute mortality
data (e.g., relating acute to chronic toxicity) and not for more subtle effects such as abnormalities, growth
or reproduction, including EC*, data (NAS, 1972). However, in the absence of other alternatives, the
0.01 application factor should be applied to EC^ data although it is recognized that these results will be
conservative and that derivation  of this historic application factor was  largely  a matter of "best
professional judgement" by the NAS (1972). Thus, site-specific review may be required in some cases
to determine compliance.
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11.2           Her III: Benthic Toxicity Tests

Toxicity tests with whole sediment are designed to determine whether the dredged material is likely to
produce unacceptable adverse effects on benthic organisms. In acute tests, the test animals are exposed
to the test sediment for 10 days and the number of survivors is recorded.


11.2.1         Species Selection

Species representing three life history strategies are recommended for use in the whole sediment toxicity
tests, one each representing a filter feeder, deposit feeder and a burrowing organism where possible
(Table 12). The rationale for testing more than a single species is to cover the range of differing species
sensitivities and to be environmentally protective. No single species is adequately protective of the broad
range of possible chemical contaminants nor of the equally broad range of possible biological responses.
Of the  species tested, at least one sensitive  benchmark (starred) species  must be used in  all cases;
however, this does  not preclude the  use of benchmark species representative of all  three required
categories. If only two different species are being tested they should, together, cover the following three
life history strategies: filter feeder, deposit feeder,  burrower.  Since amphipods are excellent organisms
for short term toxicity, they are recommended as one of the species to be tested. Non-benchmark species
listed in Table 12 can be used if a summary  of test conditions and test acceptability criteria  similar to
the starred  benchmark species are established and data from reference toxicity tests  (see  Appendix
G.3.17.2) are provided on the sensitivity of the species. Species proposed for  use regionally and not listed
in Table 12 must meet  the species  characteristics criteria provided later in  this section and proponents
must provide the following supporting information:

        •      data from toxicity tests using a set of reference chemicals with differing modes of action
               demonstrating that the proposed species is as sensitive or more sensitive than the species
               in Table 12

        •      summary of test conditions and test acceptability criteria.

Note that regions cannot substitute all species in Table 12 but that, for species used regionally in addition
to the minimum requirements, the  above supporting information is desirable but not required.

Benthic organisms are used to evaluate the potential benthic impact of dredged material disposal. Testing
of contaminated  sediments  (e.g., Word et al., 1989; Gentile et al., 1988;  Rogerson et al., 1985) and
regulatory program experience since 1977 under the Marine Protection, Research, and Sanctuaries Act
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Table 12.      Candidate Acute Toxicity Test  Species for Determining Potential Benthic Impact  of
               Dredged-Material Disposal. Details of testing procedures are provided in Appendix E.
 Amphipod Crustaceans
   Ampelisca abdita* (N)* [d,b]
   Rhepoxynius abronius* (N) [d,b]
   Grandidierellajaponica (N) [d,b]
   Corophium sp. (N) [f,d,b]
   Leptocheirus plumulosus* (E,N)" [d,b]
   Eohaustorius estuarius* (E)  [d,b]
   Hyalella azteca* (£,?)• [d,b]

 Polvchaetes
   Neanthes arenaceodentata (N)" [d,b]

 Juvenile Bivalves (clams)
   Paper pondshell clam,  Anodonta imbedllis (F)
                                     [f,b]
            other than Amphioods
 Mysid shrimp, Mysidopsis sp. (N) [f,d]
                Neamysis americana (N) [f]
                Holmesimysis costata (N) [f]
 Commercial shrimp, Penaeus sp. (N) [d,b]
 Grass shrimp, Pdaemonetes sp.  (N,E)k [d]

Insect Larvae
 Midges,   Chironomus tentans*  (F)" [d,b]
            C.  riparius * (F)* [d,b]
 Mayfly, Hexagenia limbata (F) [d,b]

Oligochaetes
 Prori/w teirfyi (F) [d,b]
 Tubifex tubifex (F)« [d,b]
 Lumbriculus variegatus (F)" [d,b]
Note:  Examples are not presented in order of importance; however, the asterisks  indicate sensitive
       recommended benchmark species. Benchmark species comprise a substantial database, represent
       the sensitive range  of a variety of ecosystems, and provide comparative data on the  relative
       sensitivity of local test species. Other species may be designated in future as benchmark species
       by EPA and the USACE when the data on their response to contaminants are adequate. Only
       benthic species should be tested. Although sediment dwellers are preferable, intimate contact with
       sediment is acceptable. Note that testing with all recommended taxa is not required; however, at
       least one starred amphipod taxon must be tested (ASTM,  1991f,g).
                            *
[f = filter feeder; d = deposit feeder; b = borrower]. Note that A.  abdita, L. plumulosus, C. tentans,
and H. limbata are not direct filter feeders, but are suspension feeders.

*      These species can also be used in sublethal, chronic testing (methods for such testing are available
       but not detailed in this manual).

b      This species can  be used  in estuarine waters down to appropriate low levels  of salinity (see
       Appendix E).

For the purposes of this manual, related to  the tolerances of the test animals, (F) = Freshwater, salinity
 «£ l%o  (N)  = Near Coastal, salinity  ^ 25%o  (E) = Estuarine, salinity  1-25%o. It  is recognized that
the commonly accepted salinity range for estuaries is l-35%o and near coastal  water  is usually greater
than 30%o salinity.
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and the Clean Water Act have shown that different species have various degrees of sensitivity to the
physical and chemical composition of sediments.

To accurately evaluate potential benthic impact, appropriately sensitive toxicity test species should be
related as closely as possible, both phylogenetically and ecologically, to benthic organisms in the disposal
site area. Commercially important but possibly less sensitive benthic species hi the vicinity of the disposal
site may also be considered for testing.

Sediment grain size is likely to vary substantially between the dredged material, the reference sediment,
and the control sediment. If candidate test species  are overly sensitive to the different grain sizes (for
instance, excessive mortality in the reference sediments attributable to grain size and not to other factors),
either this must be taken into account (e.g., DeWitt et al., 1988) or other, more grain-size tolerant species
should be considered for the project.

Final selection of test species for a particular dredged material disposal project should be made in
consultation with regional regulatory and scientific personnel. Two phylogenetically and ecologically
different species are recommended to account for different  sensitivities to contaminants. The following
is a list, not necessarily in order of importance, of characteristics to consider for species selection:

   •   readily available year-round
   •   preferably ingest sediments
   •   tolerate grain sizes of dredged material and control and reference sediments equally well or
        differences should be accounted for
   •   give consistent, reproducible response to toxicants
   •   tolerate handling and  laboratory conditions
   •   related phylogenetically and/or by ecological requirements to species characteristic of the benthic
        environment of the disposal site area in the season of the proposed disposal
   •   standardized test protocols  are available
   •   important ecologically, economically, and/or recreationally
   •   appropriately sensitive.

Infaunal amphipods are excellent organisms for short term toxicity tests with whole sediment (Swartz et
al., 1979, 1985; Mearns and  Word, 1982; Rogerson et al.,, 1985; Nebeker et al.,  1984; Gentile et al.,
1988; Scott and Redmond,  1989; Word et al., 1989; Burton, 1991),  and are strongly recommended as
appropriate test species for  acute toxicity bioassays in marine/estuarine/fresh waters.  Guidance on
available testing procedures (static, 10-d exposures) provided in ASTM (1991f,g) may be followed on
all points that do not conflict  with this manual.  Infaunal amphipods are:
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    •  sensitive
    •  readily available
    •  as a group, tolerant of a wide range of grain sizes and laboratory exposure conditions
    •  ecologically relevant to most dredged material disposal sites.

The identity of all species should be verified by experienced taxonomists, particularly for animals col-
lected in the field. If the toxicity test animals are also to be used hi estimating bioaccumulation potential,
the factors discussed hi Section 12.1.1 for species selection should also be considered.
11.2.2         Laboratory Procedures

General Test Procedures
Acceptable water quality parameters during testing include but are not necessarily restricted to:

    •   the correct temperature and pH range
    •   adequate oxygen levels
    •   proper lighting
    •   the correct salinity range (near coastal and estuarine organisms)
    •   the correct hardness range (fresh water organisms)
    •   the absence of, or insignificant concentrations of, toxicants such as ammonia.

Amphipod and other small organism tests are often, but not always, conducted in 1 L containers under
static conditions (Appendix E). Static renewal or even flow-through methods such as those described by
Redmond et al. (1989) or Benoit et al. (1993) may be required for certain tests or where static conditions
would result in unacceptable build-up of, for instance, ammonia and/or sulfides. Tests with large aquaria
(^20 L) are recommended for larger species, and should be run under continuous-flow conditions with
no more than 90% of the water volume replaced every 6-12 h. If a continuous-flow water supply is not
available, a static-renewal design is acceptable. In the latter case, no more than 75% of the water in each
exposure  chamber should be renewed 1 h before and 48 h after test  initiation and  at 48 h intervals
thereafter. Care should be taken to minimize resuspension of the sediments during these water changes.
The water should be changed more frequently if acceptable water quality cannot be maintained.

Before use, all glassware should be washed with detergent, rinsed with acetone, five times with tap water,
placed in  a clean 10% HC1 acid bath for a minimum of 4 h, rinsed five times with tap water, and then
thoroughly  flushed with either distilled or deionized water. Equipment and facilities  must provide
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acceptable lighting requirements and temperature control. An environmental incubator or a water-bath
system that allows temperature control within ±  1 °C is recommended.

Dilution water should not be stressful to the test organisms, and should be stable throughout the exposure
period.  Salinity for marine/estuarine organisms should be stable within ± 2%o and, for all organisms,
temperature should  be stable  within ±  2°C  throughout the exposure period. Dissolved oxygen
concentration should not be allowed to fall below an absolute minimum of 40% saturation for warm water
species and 60% for cold water species. The flow to the exposure chamber should be directed to achieve
good mixing without disturbing the sediment on the bottom of the chamber.

A minimum of five  replicate exposure chambers for the dredged material, reference, and control is
recommended. The standard test duration is 10 d.

The quantity of sediment needed depends on the size of the exposure chambers. The sediment should be
deep enough to meet  the biological needs of the test organisms, i.e.,  allow organisms to burrow in their
normal position, etc. Overcrowding of organisms must be avoided.

Prior to use in toxicity tests, sediments must be thoroughly homogenized. Very small amounts of clean
diluent water may be added to facilitate mixing. If separation  into liquid and solid phases occurs in
posthomogenization storage, remixing will be required prior to usage.

The reference and control sediments, as well as the dredged material being tested,  may contain live
organisms. Macrobenthic organisms should be removed by press-sieving the sediments through a 1-mm-
mesh screen. The material remaining on the screen should be noted  and discarded. The sieved dredged
material is returned to its storage container and held at 4°C. The sieved sediments should be used as soon
as practical after the macroinvertebrates are removed.

The experimental procedure described in ASTM  (199If) should be followed for preparing the exposure
chambers for amphipod toxicity tests. For larger exposure chambers, sediment should be placed on the
bottom of the exposure chamber and covered with clean diluent water; any sediment suspended during
placement should be allowed to settle for 24 h before introducing the test organisms. In continuous-flow
tests, the flow should be established after most of the suspended sediment has settled, usually 12 to 24
h, but at least 1 h before introducing the test organisms.

During the exposure period, daily records should be kept of obvious mortalities, emergence of infaunal
organisms, formation of tubes or burrows, and any other or unusual behavior. Daily records of water
quality (e.g., dissolved oxygen, salinity (if appropriate), ammonia,  temperature, pH) should be main-
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tained. In flow-through or static-renewal systems, water quality may be kept within acceptable bounds
by increasing the flow rate or frequency of water changes.

After the exposure period, live organisms are removed to clean diluent water, which may include sieving
the sediments, and then counted. If greater than acceptable mean mortality occurs in the control, as
defined in the procedures for proper conduct of mat test, the test must be repeated. Organisms which
show any response to gentle probing of sensitive parts or  gentle swirling of the  water should be
considered alive. Sediment dwellers (e.g., amphipods)  not recovered at the end of the test have to be
considered dead. If organisms from these toxicity tests are to be used in estimating bioaccumulation
potential, the survivors are gently and rapidly counted and then treated as described in Section 12.

Ammonia and Sulfide Toxicity
In order to identify elutriate or solid phase dredged material toxicity due to ammonia, it is essential to
make routine measurements of ammonia on appropriate test fractions. These measurements are compared
to water-only toxicity data for the same species used in the dredged material test (see Appendix F). The
water-only toxicity data generated separately should be generated under conditions (e.g., pH, test length)
reasonably similar to those in the test with the dredged material. If ammonia concentrations are too  low
to have potentially caused the observed toxicity in the dredged material sample, other  contaminants are
responsible for the toxicity. If ammonia concentrations are high enough  to have caused the observed
toxicity, toxicity identification evaluation (TIE) procedures should be used to confirm this suspicion.
When there is no TIE confirmation that ammonia is responsible for sediment toxicity, it must be assumed
that persistent contaminants other than ammonia are causing toxicity. Full details of procedures to identify
ammonia as a toxicant in toxicity tests with dredged material are provided in Appendix F.

Whenever chemical evidence of ammonia is present at lexicologically important levels, i.e. ammonia
concentrations exceed the species-specific acceptability ranges  shown below (or 20 mg/L for freshwater
organisms),  and ammonia is not a contaminant of concern at the disposal site, the laboratory analyst
should set up one or more beakers explicitly for the purpose of measuring interstitial ammonia. Ammonia
in the sediment interstitial water should be reduced to below the species-specific level  shown below (or
to below 20 mg/L for freshwater  organisms) before  adding the benthic  test organisms. Ammonia
concentrations in the interstitial water can be reduced by  sufficiently aerating the sample at saturation and
replacing two volumes of water per day. The analyst should measure interstitial ammonia each day until
it reaches a concentration below the appropriate species-specific level (or below 20 mg/L for freshwater
organisms).  After placing the test organisms in the sediment, the analyst should ensure mat ammonia
concentrations remain within an acceptable range by conducting the toxicity test with continuous flow or
volume replacement not to exceed two volumes per day. Peer-reviewed papers that deal with this issue
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include: Dewitt et al. (1988), Scott and Redmond (1989), Burton (1991), ASTM (1991e), EPA (1992),
Benoit et al. (1993), Ankley et al. (1991, 1992a, 1992c, 1994).
   Acceptability Ranges for Ammonia in Marine and Estuarine Amphipod Sediment Toxicity Tests.
Parameter
Ammonia (total mg/L, pH 7.7)
Ammonia (unionized mg/L, pH 7.7)
Rhepoxynius
<30
<0.4
Ampelisca
<30
<0.4
Eohaustorius
<60
<0.8
Leptocheirus
<60
<0.8
The chemistry and toxicology of sulfides is less well-understood than that of ammonia. However, sulfides
are not likely to be a problem in most open-water situations, or in  bioassays where adequate oxygen
levels are maintained in the overlying water.
11.2J
Chronic/Sublethal Tests
Chronic/sublethal responses to sediment are presently only available, in addition to the end-point of
survival, for a very few toxicity tests, for example: the amphipods Hydella azteca, Ampelisca abdita and
Leptocheirusplumulosus; the midges Chironomus tentans and C. riparius; the oligochaetes Tubifex tubifex
and Lumbriculus variegatus, and the polychaete Neanthes arenaceodentata. However, unlike acute toxicity
tests, there is presently no consensus as to what level of chronic/sublethal effects (e.g., reduction of
growth,  reproduction, fecundity, survival of young) is  cause  for concern. Further, there is also no
consensus as to when such effects would preclude disposal or would constitute unacceptable adverse
effects requiring some type of management action. Hence, chronic/sublethal tests are not presently part
of Tier  III  in this  national  manual.  However, regional testing  manuals  may apply  appropriate
chronic/sublethal tests to sediments in advance of their inclusion in this national manual provided this is
done with a benchmark species or in addition to the required benchmark testing.

Guidance for conducting the above tests may be found  in publications including  Nebeker and Miller
(1988), Nebeker et al. (1984),  Johns and Ginn (1990), Johns et al. (1990), ASTM (199le), Ingersoll and
Nelson (1990), Dillon et al. (1993), Phipps et al.  (1993), McGee et al. (1993). Burton (1991) provides
a comprehensive review of freshwater sediment  toxicity tests. Survival and growth are the endpoints of
all of these tests. In addition, some tests also measure reproductive end-points.
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Criteria for control acceptability for chronic/sublethal tests are specific to the test and organism. If control
criteria are exceeded, the test must be repeated.
11.2.4         Data Presentation and Analysis

Data Presentation
The data for each  test species should be presented in separate tables  that include the  following
information:

    •   scientific name of the test species
    •   number of organisms in each treatment at the start of the test
    •   number of organisms recovered alive and/or in normal health from each chamber at the end of
        the test (including positive and negative controls)
    •   information regarding emergence, burrowing,  tube building, behavioral abnormalities, growth,
        reproduction, and any other observations
    •   water-quality data for each test chamber for each day.

Data Analysis
It is possible that neither mortality nor other effects will be observed in any of the treatments or that
survival in the dredged material will be equal  to or higher than survival  in the reference or control
sediments. In either of these situations, there is  no need for statistical analysis and no indication of
adverse effects due to the dredged material. Similarly, if survival is higher in test sediments than in the
control, but lower than in the reference area, and control survival  is at acceptable levels (i.e., 90% or
greater survival), there is no need for statistical analysis and no indication of benthic toxicity due to the
dredged material. However,  if survival in the reference sediment is higher than in the dredged material
treatments and exceeds the allowable percent difference between the two treatments,  the data have to be
analyzed statistically to determine  whether there is a significant difference between the reference and
dredged material. Statistical procedures recommended  for analyzing benthic  acute  toxicity data  are
described in detail in Appendix D. Local guidance must be developed to interpret chronic/sublethal tests.
11.2.5         Conclusions

Guidance on the use of the results to reach a determination is provided in Section 6.2.
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11J           Her IV: Chronic/Sublethal Effects Evaluations

At present, it is not appropriate to incorporate sediment chronic/sublethal effects testing in this national
manual (see Sections 6.0 and 11.2.3). When standardized chronic effects tests are approved, they will
be incorporated in Tier HI. Until then, such non-standard tests should be used in Tier IV except where
regional testing manuals apply such tests in advance of their inclusion in future revisions of this national
manual, provided this is done with a benchmark series or in addition to the required testing.
11.4           Tier IV: Case Specific Evaluations

Biological effects tests in Tier IV should be used only in situations that warrant special investigative
procedures. They may include chronic/sublethal tests, field studies such as benthic infaunal studies (EPA,
1992), experimental studies such as in situ toxicity tests or toxicity identification evaluation (Ankley et
al., 1992a), risk assessments and/or no effects levels for aquatic life. In such cases, test procedures have
to be tailored for specific situations, and general guidance cannot be offered. Such studies have to be
selected, designed, and evaluated as the need arises, with the assistance  of administrative and scientific
expertise from EPA and USAGE, and other sources as appropriate.
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                                      144
                   EVALUATE EXISTING
                 INFORMATION; (POSSIBLE
                    LIMITED TESTING
                   FOR EXCLUSIONS)
    WATER COLUMN
      BENTHOS
    MEASURE AND
  MODEL DISSOLVED
   CONTAMINANTS;
  COMPARE TO WQS
CALCULATE THEORETICAL
   BIOACCUMULATION
  POTENTIAL; COMPARE
    TO REFERENCE
  MEASURE TOXICITY;
  MODEL SUSPENDED
  PHASE; DETERMINE
TOXICITY AFTER MIXING
  MEASURE TOXICITY;
      MEASURE
COMPARE TO PDA UMTTS
  AMD TO REFERENCE
      CONDUCT
   CASE-SPECIFIC
   TOXICITY TESTS
      CONDUCT
    CASE-SPECIFIC
      TOXICITY;
                                      OTHER TESTS
                                  TIER I
                          (GENERALLY REPRESENTS
                           EXISTING INFORMATION)
     TIER II
(SOLELY CONCERNED
 WITH CHEMISTRY)
     TIER III
 (GENERIC BIOASSAY
   [TOXICITY AND
 BIOACCUMULATION]
      TESTS)
     TIER IV
 (SPECIFIC BIOASSAY
   [TOXICITY AND
 BIOACCUMULATION]
 AND OTHER TESTS)
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12.0          GUIDANCE FOR PERFORMING BIOACCUMULATION TESTS

Bioaccumulation is defined in relation to disposal activities in the Definitions section at the beginning of
this manual.


12.1          Tier III:  Determination Of Bioavailability

Unavailability tests  are  designed to evaluate  die  potential of benthic organisms to bioaccumulate
contaminants of concern from the proposed dredged material. Lee et al. (1989) and Boese and Lee (1992)
discuss bioaccumulation methodology in detail and may be followed on any matter that does not conflict
with this manual. Tier III bioavailability tests are based on analysis of tissues of organisms after 28 d of
exposure (see Section 6.3).  Although time series testing is a component of Tier IV bioaccumulation
testing, it may also be appropriate in Tier III, for instance where K^ values are greater than 5.5 (see
Section 12.2.1).


12.1.1        Species Selection and Apparatus

The selection of aquatic organisms for use in the determination of bioaccumulation will depend on their
inability to metabolize some types of organic compounds, and their ability to survive exposure to the test
sediments. Two  species should be used in bioaccumulation testing where possible (Table  13), unless
adequate regional data are available to justify single species testing. Test species should provide adequate
biomass for chemical analysis, and preferably  ingest sediments and survive in dredged material and
control and reference sediments equally well (or where differences can be accounted for). The rationale
for testing more than a single species is to cover the range of differing species contaminant accumulation
and to be environmentally protective. Of the species tested, at least one must be a benchmark species;
however, this does not preclude the use of more than one benchmark species. Non-benchmark species
listed in Table 13 can achieve benchmark status if a summary of test conditions and test acceptability
criteria similar to the starred benchmark species are provided that meet the required species characteristics
criteria.  Species proposed for use regionally and not listed in Table 13 must also meet  the species
characteristics criteria and proponents must provide  a summary of test conditions and test acceptability
criteria except where species are to be tested in addition to the minimum requirement. In this latter case,
this information  is desirable but not required.
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Table 13.     Candidate Test Species for Determining Potential Bioaccumulation from Whole Sediment
              Tests. Details of testing procedures are provided in Appendix E.
 Polychaetes
        Neanthes arenaceodentata* (N)
        Nereis rtrens* (N.E)1
        Arenicda marina (N)
 Oligochaetcs
        Lumbrictdus variegatus (F)*

 Insect Larvae
    Mayfly, Hexagenia limbata or sp. (F)
Bivalves
  Macoma clam, Macoma 7uuufa*(N,E)*
  Yoldia clam, Yoldia ttmatula (N)   •
Crustaceans
  Diporeia sp. (F)
Note:  Examples are not presented in order of importance; however, the asterisks indicate recommended
       benchmark species. Other species may be designated in future as benchmark species by EPA and
       USACE when the data on their response to contaminants  are adequate. Only benthic species
       should be tested. Although sediment ingesters are preferable, intimate contact with sediment is
       acceptable.

       Only tests which do not require feeding of the organisms  are included. Feeding is a research
       issue; for the present, food is not to be added because it provides additional organic carbon and
       can alter contaminant partitioning during testing.

For the purpose of this manual, related to the tolerances of the test  animals, (F)  = Freshwater, salinity
^ l%o (N)  = Near Coastal, salinity  ^ 25%o  (E) = Estuarine, salinity 1-25 %o.  It is recognized that
the commonly accepted salinity range for estuaries is l-35%o and near coastal water is usually greater
than 30 %c  salinity.

   *   Macoma nasuta and Nereis virens bioaccumulation tests are in the process of standardization by
       EPA; it is expected that these will, hi future, be the primary benchmark species for near coastal
       waters.  Further, these two species can be used  in estuarine waters down to appropriate low
       levels of salinity (see Appendix E).
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Apparatus to be used for testing is described in Section 11.2.2. Additional requirements for voiding gut
contents are described in Section 12.1.2. Species characteristics to consider when designing  bio-
accumulation tests include, not in order of importance:

    •   readily available year-round
    •   provide adequate biomass for analysis
    •   preferably ingest sediments
    •   survive in dredged material and control and reference sediments equally well, allowing adequate
        tissue for analysis
    •   tolerate handling and laboratory conditions
    •   related phylogenetically and/or by ecological requirements to species characteristic of the disposal
        site area in the season of the proposed discharge
    •   important ecologically, economically, and/or recreationally
    •   inefficient metabolizers of contaminants, particularly PAH.

Regional scientists and regulatory personnel should be consulted  for additional guidance. A minimum
amount of tissue is  required for  analysis, otherwise it will be impossible to quantify the amount of
contaminant present (Section 9.S.2). Examples  of the amounts of tissue which may be required are
provided in Table 4. However, the amounts shown are not set amounts; more or less may be required
depending on the analytes, matrices, detection limits, and particular analytical laboratory. If the biological
needs  of the  organisms  or adequate  voiding  (e.g.,  clams)  require  the  presence  of sediment,
uncontaminated sand should be used (Section 12.1.2). Data in the form of "concentration below detection
limits" are not quantitative; definitive concentration measurements are the goal, where such are possible
within reasonable method and target detection limits.
12.1.2         Experimental Conditions

Test conditions are similar to those described  in Section  11.2.2 for whole sediment toxicity tests.
Overlying water renewal may be required to maintain adequate water quality. Food or additional sediment
should not be provided during the test. Control animals should be sampled and archived at both the
beginning and the end of testing. If discrepancies are found during data analysis, the archived samples
can be analyzed to  possibly resolve any problem(s). Due care should be taken not to exceed species-
specific biomass loadings (overcrowding; APHA, 1989).

Digestive tracts of the  animals should be emptied or removed immediately  after termination of the
exposure period. Sediment in digestive tracts may contain  inert constituents and  the contaminants of
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concern in forms which are not biologically available but which may be incorrectly identified as such
during chemical analysis (e.g., see Lobel et al.,  1991).

If the animals are large enough to make it practical, the best procedure is to excise the digestive tract.
However, test organisms are seldom large enough to allow this, and most organisms have to be allowed
to void the material, in separate aquaria in clean, sediment-free water. Some organisms will pass material
through the digestive tract only if more material is ingested. These animals have to be purged in aquaria
with clean sand.  Animals are not fed during the purging period. Fecal material is siphoned from the
aquaria twice during the 24-h purging period. To minimize the possibility of loss of contaminants from
tissues, purging for longer periods is not recommended. Shells or exoskeletons which generally contain
low levels of contaminants are, where possible, removed and not included in the analysis as their weight
would give an artificially low indication of bioavailability.

An initial time-zero of each sample is collected for tissue analysis. Tissue contaminant concentrations in
control  animals must  be determined  to  ensure that background levels  are not inordinate.  Although
procedures for Tier III and IV laboratory bioaccumulation tests have been discussed separately, it may
be possible to combine these procedures in practice.  This can be done by following the steady state (Tier
IV) bioaccumulation procedure which involves sequential time-series analyses, but initially analyzing only
the 28 d sample  and  freezing the other samples. If these data, as part of the Tier III bioavailability
evaluation, do  not allow a determination to be made, then the remaining time series samples may be
analyzed and used in the Tier IV steady-state bioaccumulation evaluation.
12.1.3         Chemical Analysis

Chemical analysis will  involve some or all of the contaminants identified in Sections 4.2 and 9.5.1.
Analytical procedures are provided in Section 9.S.2.
12.1.4         Data Presentation and Analysis

Data Presentation
Data should be presented in tabular format, listing tissue concentration of each contaminant, by organism
and by sediment type (e.g., dredged and reference). Similar information to that detailed in Section 11.2.4
should  be provided. Although bioaccumulation species/tests  cannot be used  to  determine toxicity
requirements, any mortalities which occur during bioaccumulation testing must be documented.
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Data Analysis
Contaminant tissue concentrations in test organisms are statistically compared to the FDA Action Levels
(Table 2) (refer to Figure 3). These tissue concentrations are also statistically compared with reference
organism concentrations (Appendix D). In some cases, tissue concentrations in organisms exposed to one
or more of the dredged-material samples may be less than or equal to reference organism concentrations.
Providing the reference data are appropriate, this result indicates that bioavailability of the contaminants
of concern in the dredged material is not greater than in the reference area sediment.

The sample of organisms archived at the initiation of the exposure can be useful in interpreting results.
It can add perspective to the magnitude of uptake during the exposure period. In some  cases, elevated
body burdens may not be due to the dredged material or reference sediment, but may have been already
present in the organisms at the start of the test.
12.1.5         Conclusions

Guidance on reaching a determination is provided in Section 6.3.


12.2           Tier IV: Determination Of Steady State Bioaccumulation

Tier IV bioaccumulation evaluation, if necessary, provides for determination, either by laboratory testing
(ASTM,  1984) or by collection of field samples, of the steady state concentrations of contaminants in
organisms exposed to the dredged material  as compared with organisms exposed to the reference site
material. Testing options include longer laboratory exposures (not discussed), collection of organisms
living in the material to be dredged and at the reference site for body burden determinations (Section
12.2.2) or in situ exposures using transplanted organisms, for instance caged mussels (not discussed). Tier
IV determinations follow the guidance in Section 7.2.


12.2.1         Laboratory Testing

The necessary species, apparatus and  test conditions  for laboratory testing  are  those  for Tier III
bioaccumulation testing (see Sections 12.1.1 and 12.1.2). Tissue samples taken at different times during
the exposure period provide the basis for determining the rate of uptake and elimination of contaminants.
From these rate data, the steady state concentration of contaminants in the tissues can be calculated, even
though the steady state might not have been reached during the actual exposure. For the purposes of this
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test, steady state is defined as the concentration of contaminant that would occur hi tissue after constant
exposure conditions.

An initial time-zero sample of each species is collected for tissue analysis. Additional tissue samples are
collected from each of the five replicate reference and dredged-material exposure chambers at intervals
of, for instance, 2,4,7,10,18, and 28 d. It is critical that enough tissue is available to allow for interval
body burden analyses at the specified detection limits.

Complete tissue concentration data  should  be presented in tabular format.  Recommended  statistical
methods for fitting a curve to determine steady-state tissue concentration are provided in Appendix D.
The statistical procedures use an iterative  curve-fitting process to determine the key variables (k^C. the
uptake rate-constant tunes the contaminant concentration in the sediment, and ^ the depuration rate con-
stant).  An initial value for C. has to be supplied. When the sediment concentration of the contaminant of
concern is used, the ratio  of kjk^ is the  sediment bioaccumulation factor (BAF) (Lake et al., 1987;
Rubinstein et al., 1987),  the ratio of steady-state tissue concentration to sediment concentration.

A determination is made based  on the magnitude of bioaccumulation from the dredged material,  its
comparison with the available FDA levels,  steady-state bioaccumulation from the reference sediment, and
the body burden of reference organisms. Guidance for making determinations based on these comparisons
is provided in Section 7.2 and can include risk assessment and no effects levels for aquatic life.

Guidance on quality assurance/quality  control (QA/QC) considerations for  bioaccumulation testing are
provided hi Appendix G.3.17 and EPA (1994b).
12.2.2         Field Assessment of Steady State Bioaccumulation

Field sampling programs obviate difficulties related to quantitatively considering field-exposure conditions
in the interpretation of test results, since the animals are exposed to the conditions of mixing and sediment
transport actually occurring at the disposal site. Difficulties related to the time required to conduct labora-
tory bioaccumulation studies are also overcome if organisms already living at the disposal site are used
for field bioaccumulation studies. This approach is technically valid for predictive purposes only where
there is a true historical precedent for the proposed operation being evaluated. That is, a field assessment
can be used only where the quality of the sediment to be dredged can be shown not to have deteriorated
or become more contaminated since the last dredging and disposal operation. In addition, disposal has
to be proposed for the site at which the dredged material in question has been previously disposed or for
a site of similar sediment type supporting a similar biological community. This approach is generally not
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appropriate for multi-user disposal sites. Knowledge of the contaminant body burden of the organisms
living around the proposed disposal site is used in evaluating bioaccumulation results in Tier IV (Section
7.2).
12.2.2.1              Apparatus

Major items required include:

    •   a vessel capable of operating at the disposal site and equipped to handle benthic sampling devices;
        navigation equipment has to allow precise positioning
    •   sampling devices such as a box corer, Smith-Maclntyre, Van Veen, Petersen, Ponar, Ekman or
        other benthic grab
    •   stainless steel screens to remove animals from the sediment
    •   tanks for transporting the animals to the laboratory in collection site water
    •   laboratory facilities for holding the animals prior to analysis
    •   chemical and analytical facilities as required for the desired analyses.
12.2.2.2              Species Selection

The species selected for analysis have to be present in sufficient numbers for adequate sample collection
at all stations and to provide sufficient tissue for analysis (see Section 12.1.1). The same species must
be collected at all stations because bioaccumulation cannot be compared  across species lines. If these
conditions cannot be met, the field assessment approach cannot be implemented.

If possible, several samples of sufficient size for analysis should be collected at each station to provide
a statistical estimate of variability in tissue contaminant content. Collection of more than one sample per
station, however, may prove impractical if a composite of many small organisms has to be  used or if
suitable organisms are not abundant at the disposal site.

To minimize the numbers and collection effort required, it is desirable to select the largest appropriate
species. However, highly mobile epifauna  (such as crustaceans, certain molluscs, and fish) should not
be used, because a relationship cannot be established between their location when collected and their
body burden at the time of collection. Therefore, relatively large, immobile species are the most desirable
organisms. However, analyses should not be conducted on single organisms as the objective is to obtain
representative data for the entire population of organisms.  Any relatively immobile species collectable
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in sufficient numbers at all stations may be used, but the required collection effort increases sharply as
organism size decreases.

As discussed previously, if PAH are contaminants of concern, it is essential that bioaccumulation studies
include one or more species with very low ability to metabolize PAH. Bivalve molluscs and oligochaetes
are widely accepted as meeting this requirement.
12.2.2 J              Sampling Design and Conduct

Sufficient tissue to obtain definitive body burden data has to be collected using the same species from
each of at least three stations within the disposal site boundaries and from an acceptable reference site.
It is mandatory that several stations be sampled, rather than collecting all of the animals at one station,
in order to provide a measure of the variability that exists in tissue concentrations in the animals in the
area. Samples from all stations should be collected on the same day if possible.
12.2.2.4              Basis for Evaluation of Bioaccumulation

Evaluations are made by comparison to contaminant concentrations in field organisms living around, but
not affected by, the disposal site, similar to the reference area approach (Section 3.1). In this case,
reference data involve at least three stations located in an uncontaminated material sedimentologically
similar to that within the disposal site, in a direction perpendicular to (i.e., not in the direction of) the
net bottom transport. If the  direction of  net bottom transport  is not known,  at least six stations
surrounding the disposal site should be established in sediments sedimentologically similar to those within
the disposal site.
12.2.2.5               Sample Collection and Handling

Repeated collections should be made at the same location until an adequate tissue volume is obtained.
Gently wash the sediment obtained by the sampler through 1-mm mesh stainless-steel screens, and place
the retained organisms of the desired species in holding tanks.

Label the samples clearly and return the organisms to the laboratory, being careful to keep them separated
and to maintain nonstressful levels of temperature and dissolved oxygen. In the laboratory, maintain them
in clean water hi separate containers. Do not place any sediment in the containers and do not feed the
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organisms. Immediately discard any organisms that die. Remove sediment from the digestive tracts of
the organisms and, as possible, shells or exoskeletons (Section 12.1.2).
12.2.2.6              Chemical Analysis

Chemical  analysis will involve some or all of the contaminants identified in Sections 4.2 and 9.5.1.
Analytical procedures are provided in Section 9.5.2.
12.2.2.7              Data Presentation and Analysis

Complete tissue concentration data for all samples should be presented in tabular format as previously
described. Since Tier IV  testing will generally use non-standard methods  and approaches,  complete
documentation is critical. Recommended statistical methods presented in Appendix D may not include all
data analyses necessary for all Tier IV tests.
12.2.2.8              Conclusions

A determination is made based on the magnitude of bioaccumulation in organisms collected within the
boundaries of the reference site, compared with bioaccumulation in organisms living within the area to
be dredged. Guidance for making a determination based on these comparisons is provided in Section 7.2.
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       Molluscs. Standard Practice No. E-1022-84. American  Society for Testing and Materials,
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       Species of Saltwater Bivalve Molluscs. Method E724-89. In: Annual Book of ASTM Standards,
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Babinchak, J. A., J. T. Graikoski, S. Dudley and M. F. Nitkowski. 1977. Effect of dredge spoil
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Ballschmiter, K. and M. Zell. 1980. Analysis of poly chlorinated biphenyls (PCB's) by glass capillary gas
       chromatography, composition of technical arochlor- and clophen-PCB mixtures. Fresenius Anal.
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       Tech.  Rept. EPA/CE-81-1 prepared by Great Lakes Laboratory, State University College at
       Buffalo,  Buffalo, NY, for the U.S. Environmental Protection Agency/U.S. Army  Corps of
       Engineers Technical Committee on Criteria for Dredged and Fill Material. Published by the U.S.
       Army Engineer Waterways Experiment Station, Vicksburg, MS.

Pratt, H. T. 1991.  Terminology management in ASTM Committees.  Standardization News December
       1991:16-19. American Society for Testing and Materials, Philadelphia, PA.
                                                                            DRAFT

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	168	

Rand, G. M. and S. R. Petrocelli. 1985. Fundamentals of Aquatic Toxicology .-Methods and Applications.
       Hemisphere Pub. Co., Washington, D.C. 666 pp.

Randall, R.C., H. Lee II and R.J. Ozretich. 1991. Evaluation of selected lipid methods for normalizing
       pollutant bioaccumulation. Environ. Toxicol. Chem.  10:1431-1436.

Rao, V.  C., K. M. Seidel, S. M. Goyal, T.  G. Metcalf and J.  L. Melnick. 1984. Isolation of
       enteroviruses from water,  suspended solids and sediments from Galveston Bay:  survival of
       poliovirus and rotavirus adsorbed to sediments. Appl. Environ. Microbiol. 48:404-409.

Redmond, M.S., K.J. Scott, K.M. McKenna and D.  Robson. 1989. Evaluation of the relative acute
       toxicity of nearshore sediments in Narragansett Bay. Report prepared by Science Applications
       International Corporation for U.S. EPA, Narragansett Bay Program Office. 27pp. + appendices.

Rice, C., F. Espourteille  and R.  Huggett.  1987.  A method for analysis  of tributyltin in estuarial
       sediments and oyster tissue, Crassostreavirginica. Appl. Organomet. Chem. 1:541-544.

Rogerson, P.P., S.C. Schimmel and G. Hoffman.  1985.  Chemical and biological characterization of
       Black Rock Harbor dredged material. Tech. Rept. D-85-9 prepared by the U.S. Environmental
       Protection Agency,  Narragansett,  RI, for the U.S. Army Engineer Waterways Experiment
       Station, Vicksburg, MS.

Rubinstein, N.I., J.L. Lake, R.J. Pruell, H. Lee, II, B. Taplin, J. Heltshe, R. Bowen and S. Pavignano.
       1987. Predicting bioaccumulation of sediment associated organic contaminants:  Development of
       a regulatory tool for dredged material  evaluation. Tech. Rept. D-87  prepared by the U.S.
       Environmental Protection Agency,  Narragansett, RI, for the U.S. Army Engineer Waterways
       Experiment Station, Vicksburg, MS. 59 pp.

SAB. 1992. Technical Review of "Evaluation of Dredged Materials Proposed for Ocean  Disposal -
       Testing Manual". Report of the Sediment Quality Subcommittee of the Ecological Processes and
       Effects Committee,  U.S. EPA Science Advisory Board, Washington, D.C. EPA-SAB-EPEC-92-
       014. 20 pp.

SAB. 1994. An SAB report: Evaluation  of a  Testing Manual for  Dredged Material Proposed for
       Discharge in Inland and Near Coastal Waters. Report of the Sediment Quality Subcommittee of
       the Ecological Processes and Effects Committee, U.S. EPA Science Advisory Board, Washington,
       DC. EPA-SAB-EPEC-94-007. 16 pp.
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	169	

SAIC. 1992a.  Survey of sediment toxicity in Long Island Sound. Final report prepared by Science
       Applications International Corporation for NOAA, Rockville, MD, under Task Order 55-DSNC-
       01-00006.

SAIC. 1992b. Survey of sediment toxicity in the Hudson/Raritan Bay estuary. Final report prepared by
       Science Applications International Corporation for NOAA, Rockville, MD, under Contract No.
       68-C8-0066.

Schindler, B. 1991. Verbal reliability. Editorial, Standardization News December 1991:1.  American
       Society for Testing and Materials, Philadelphia, PA.

Schwartz, T.R., R.D. Campbell, D.L. Stalling, R.L. Little, J.D. Petty, J.W. Hogan and E.M. Kaiser.
       1984. Laboratory data base for isomer-specific determination of polychlorinated biphenyls. Anal.
       Chem.  56:1303-1308.

Schwartz, T.R., D.E. Tillitt, K.P. Feltz and P.H. Peterman.  1993. Determination of nono- and non-O,O'
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       Chemosphere 26:1443-1460.

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       populations of the tubiculous  amphipod Ampelisca abdlta. ASTM STP 1027. American Society
       for Testing and Materials, Philadelphia, PA

Shiharis, M. P., A. C. Rex, G. W. Pettibone, K. Keay,  P. McManus, M.  A. Res, J. Ebersole and E.
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       in the estuarine environment. Appl. Environ. Microbiol. 35:685-689.

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       fractionation. Anal. Chem. 53:2152-2154.

Smith, L.M.,  D.L.  Stalling  and I.L.  Johnson.  1984.  Determination of part-per-trillion levels of
       polychlorinated dibenzofurans and dioxins in environmental samples. Anal. Chem. 56:1830-1842.
                                                                              DRAFT

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	170	

Stalling, D.L., T.R.  Schwartz, W.J. Dunn, HI and S. Wold. 1987. Classification of polychlorinated
       biphenyl residues. Anal. Chem. 59:1853-1859.

Strickland, J.D.H. and T.R. Parsons. 1972. A practical handbook of seawater analysis. Bull. Fish. Res.
       Bd. Canada 122:1-172.

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       Assessment and Remediation. National Academy Press, Washington, DC.

Swartz, R.C., W.A.  DeBen  and F.A. Cole. 1979. A bioassay for the toxicity of sediment  to marine
       macrobenthos. Water Pollut. Control Fed. 51:944-950.

Swartz, R.C., W.A. DeBen, J.K.P.  Jones, J.O.  Lamberson and  F.A. Cole. 1985. Phoxocephalid
       amphipod  bioassay for marine sediment toxicity. Pp. 284-307 In: R.D. Cardwell, R. Purdy, and
       R.C. Bahner  (Eds.),  Aquatic  Toxicology and Hazard Assessment. Seventh Symposium. ASTM
       STP 854.  American Society for Testing and Materials, Philadelphia, PA.

Tatem, H.E., D.L. Brandon, C.R. Lee, A.S. Jarvis and R.G. Rhett.  1991.  Effects of storage on
       sediment toxicity, bioaccumulation potential and chemistry. U.S. Army Corps of Engineers,
       Waterways Experiment Station Misc. Paper EL-91-2.  62  pp.

Tavolaro,  J. F. and J. M. Mansky. 1985. Effects  of dredging operations on nutrients and suspended
       sediment concentrations. Northeast Environ. Sci. 3:208-216.

Tetra Tech. 1985.  Bioaccumulation monitoring guidance:  1. Estimating the potential for bioaccumulation
       of priority pollutants and 301(h) pesticides discharged into marine and estuarine waters. Final
       report prepared by Tetra Tech,  Inc.,  for  the U.S. Environmental Protection Agency under
       Contract No.  68-01-6938.

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       Protection Agency under Contract No. 69-01-6938.
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	171	

Tetra Tech. 1986b. Bioaccumulation monitoring guidance: 4. Analytical methods for U.S. EPA priority
       pollutants and 301(h) pesticides in tissues from estuarine and marine organisms.  Final report
       prepared by Tetra Tech, Inc., for the U.S. Environmental Protection Agency under Contract No.
       68-01-6938.

Thomann, R.V. 1989. Bioaccumulation model of organic chemical distribution in aquatic  food chains.
       Environ. Sci. Technol. 23:699-707.

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       1989. Findings of tributyltin, dibutyltin and monobutyltin in bivalves from selected U.S. coastal
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       (Federal Water Pollution Control Act Amendments of 1972). Misc. Paper D-76-17, U.S. Army
       Engineer Waterways Experiment Station, Vicksburg, MS.

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       A  Technical  Framework.  EPA  842-H-92-008.  U.S.  Army  Corps  of  Engineers and U.S.
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       sediments. Water Res. 5:1079-1087.

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       and water solubility to estimate bioconcentration factors for organic chemicals in fish.  Pp.  116-
       129 la: J.G. Eaton, P.R. Parish, and A.C. Hendricks (Eds.), Aquatic Toxicology.  ASTM  STP
       707. American Society for Testing and Materials, Philadelphia, PA.

Walker, M.K., L.C. Hufnagle,  M.K. Clayton and R.E.  Peterson. 1992.  An egg injection method for
       assessing early life stage mortality of polychlorinated dibenzo-p-dioxins, dibenzofurans and
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WDNR/USGS.  1992.  Sources of pollutants in Wisconsin stormwater, April 1992. Unpublished report
       prepared by Wisconsin Department of Natural Resources and U.S. Geological Survey.
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	172	

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       amphipod bioassays. Pp. 467-473 Isi Oceans '89, Vol. 2: Ocean Pollution. Institute of Electrical
       and Electronics Engineers,  New York, NY.

Wright, T.D.  and L.H. Saunders. 1990. U.S. Army Corps  of Engineers dredged material testing
       procedures. Environ. Profess. 12:13-17.

Young, D.  1988. Report on the Assessment and Application of Pollutant Biomagnification Potential in
       Near Coastal Waters. EPA Report 600/X-88/295. U.S. Environmental Protection Agency, ERL-
       Narragansett N-065.
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                                                                                                i

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  APPENDIX A
40 CFR PART 230
                          DRAFT

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Wednesday
December 24, 1980
Part IV



Environmental

Protection Agency

Guidelines for Specification of Disposal
Sites for Dredged or Fill Material

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 65336    Federal Register / Vol. 45. No. 249 / Wednesday. December 24. 1980 / Rules and Regulations
  NVIRONMENTAL PROTECTION
  GENCY
 40 CFR Part 230

 [WH-FHL 1647-7]

 Guidelines for Specification of
 Disposal Sites for Dredged or Fill
 Material
 AGENCY: Err. Ironrrental P:o:r>  : ,jn
 \gency.
 ACTION: Rule.

 SUMMARY: The 404(b)(l) Guidelines are
 the substantive criteria used in
 evaluating discharges of dredged or fill
 material under section 404 of the Clean
 Water Act. These Guidelines revise and
 clarify the September 5. 1975 Interim
 final Guidelines regarding discharge of
 dredged or fill material into waters of
 the United States in order to:
  (1) Reflect the 1977 Amendments of
 Section 404 of the Clean Water Act
 (CWA);
  (2) Correct inadequacies in the interim
 final Guidelines by filling gaps in
 explanations of unacceptable adverse
 ir.pacts on aquatic ecosystems jnd by
 r6quTing documentation of compliance
 \% ,:h the Guidelines: and
  ( J| Produce a final rulcmaking
  FECTIVE DATE: These Guidelines will
?Tpr'!v in al! 404 permit decisions made
after M.irch 23, 1981. In the case of civil
vu,rks projects of the United States
Ai'.-i'.y Corps of Engineers involving the
c!- > harge of dredged or fill material for
uh.c". there is no permit application or
permit as such, these Guidel.nes will
app'> to all projects on which
construction or dredging contracts are
issued, or on which dredging is initiated
for Corps operations not performed
under contract, after October 1. 1981.  In
the case of Federal construction projects
meeting the criteria in section 404(r).
these Guidelines will apply to all
projects for which a final environmental
impact statement is filed with EPA after
April 1, 1981.
FOR FURTHER INFORMATION CONTACT:
Joseph Krivak. Director. Criteria and
Standards Division (WH-585).
Environmental Protection Agency. 401 M
Street. S.W., Washington. D.C. 20460.
telephone (202) 755-0100.
SUPPLEMENTARY INFORMATION:

Background
  The section 404 program for the
evaluation of permits for the  discharge
of dredged or fill material was originally
  acted as part of the Federal Water
  llution Control Amendments of 1972.
  e section authorized the Secretary of
 the Army acting through the Chief of
 Engineers to issue permits specifying
 disposal sites in accordance with the
 section 404(bj(l) Guidelines. Section
 404(b)(2) allowed the Secretary to issue
 permits otherwise prohibited by the
 Guidelines, based on consideration of
 the economics of anchorage and
 navigation. Section 404(c) authorized the
 Administrator of the Environmental
 Protection Agency to prohibit or
 withdraw the specification of a site.
 upon a determination that use of the site
 would have an unacceptable adverse
 ef feet on municipal water supplies.
 shellfish beds and fishery areas
 (including spawning and breeding
 areas), wildlife, or recreational areas.
  Under section 404(b)(l),  the
 Guidelines are to be based on criteria
 comparable to those in section 403(c) of
 the Act. for the territorial seas.
 contiguous zone, and oceans. Unlike
 403(c). 404 applies to all waters of the
 United States.  Characteristics of waters
 of the United States vary greatly, both
 from region to  region and within a
 region. There is a wide range of size,
 flow, substrate, water quality, and use.
 In addition, the materials to be
 discharged, the methods of discharge.
 and the activities associated with the
 discharge also vary widely. These and
 other variations make it unrealistic at
 this time to arrive at numerical criteria
 cr standards for toxic or hazardous
 substances to be applied on a
nationwide basis. The susceptibility of
 the  aquatic ecosystem to degradation by
 purely physical placement of dredged or
 fill material further complicates the
 problem of arriving at nationwide
 standards. As a result, the Guidelines
concentrate on specifying the tools to be
used in evaluating and testing the
impact of dredged or fill material
discharges on waters of the United
States rather than on simply listing
numerical pass-fail points.
  The first section 404(b)(l) Guidelines
 were promulgated by the Administrator
 in interim final form on September 5,
1975, after consultation with the Corps
of Engineers. Since promulgation of the
interim final Guidelines,  the Act has
been substantially amended. The Clean
Water Act of 1977 established a
procedure for transferring certain
permitting authorities to  the states,
exempted certain discharges from any
section 404 permit requirements, and
gave the Corps enforcement authority.
These amendements also increased the
 importance of the section 404(b)(l)
Guidelines, since some of the
exemptions are based on alternative
ways of applying the Guidelines. These
changes, plus the experience of EPA and
the Corps in working with the interim
final Guidelines, have prompted a
revision of the Guidelines. The proposed
revision attempted to reorganize the
Guidelines, to make it clearer what had
to be considered-in evaluating a
discharge and what weight should be
given to such considerations. The
proposed revision also tightened up the
requirements for the perrr.ittir.g
authority s documentation of the
application of the Guidelines.
  After extensive consultation w:th the
Corps, the proposed revisions were put
out for public comment (44 FR 54222.
September 18. 1979). EPA has reviewed.
and. after additional consultation with
the Corps, revised the proposal in light
of these comments. This preamble
addresses the significant comments
received, explains the changes made in
the cegulation,  and attempts to clear up
some misunderstandings which were
revealed by the comments. Response to
Significant Comments

Regulation  Versus Guideline

  A number of commenters objected to
the proposed Guidelines on the grounds
that they were too "regulatory." These
commente.-: ..-gued that the term
"guidelines" which appears in section
404(b)(l) requires a document with less
binding effect than a regulation. EPA
disagrees. The Clean Water Act does
not use the word "guideline"  to
distinguish advisory information from
regulatory requirements. Section
404(b)(2) clearly demonstrates that
Congress contemplated that discharges
could be "prohibited" by the  Guidelines.
Section 403 (which is a model for the 404
(b)(l) Guidelines) also provides for
"guidelines" which are clearly
regulatory in nature. Consequently, we
have not changed the regulation to make
it simply advisory. Of course, as the
regulation itself makes clear, a certain
amount of flexibility is still intended.
For example, while the ultimate
conditions of compliance are
"regulatory", the Guidelines  allow some
room for judgment in determining what
must be done to arrive at a conclusion
that those conditions have or have not
been met. See. for example, § 230.8 and
§ 230.60,  and introductory sentence in
§ 230.10.

Statutory Scheme and How the
Guidelines Fit Into It

  A number of commenters with
objections appeared confused about
EPA's role in the section 404 program.
Some wondered why EPA was issuing
Guidelines since EPA could stop an
unacceptable discharge  under section
404(c). Others were uncertain how the

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         Federal  Register / Vol. 45. No.  249 / Wednesday. December 24. 1980 /  Rules and Regulations  85337
 Guidelines related to other section 404
 regulations.
   The Clean Water Act prohibits the
 discharge of dredged or fill material
 except in compliance with section 404.
 Section 404 sets up a procedure fur
 issuing permits specifying discharge
 sites. Certain discharges (e g. emergency
 repairs, certain farm and forest roads.
 and other discharges id>fi!if.»d in
 sections 404(f) •'nd (r))  *rc exempted
 from the permit requirements. The
 permitting authority  (either the Corps of
 Engineers or an approved State
 program) approves discharges at
 particular sites through application of
 the section 404(b)(l)  Guidelines, which
 are the substantive criteria for dredged
 and fill material discharges under the
 Clean Water Act. The Corps also
 conducts a Public Interest Review.
 which ensures that the  discharge will
 comply with the applicable
 requirements of other statutes and be in
 the public interest. The Corps or the
 State, as the case may be. must provide
 an opportuni'y for a public hearing
 before making its decision whether to
 approve or deny. If the  Corps concludes
 that the discharge does not comply with
 thn Guidelines, it may still issue the
 permit under 404(b)(2) if it concludes
 that the economics of navigat:on and
 anchorage warrant. Section 404(b)(2)
 Rives the Secretary a limited authority lo
 issue permits prohibited by the
 Guidelines; it does not. as some
 coinmer.ters suggested, require the
 Guidelines to consider the economics of
 navigation and anchorage. Conversely,
 because of 404(b)(2).  the. fact that a
 discharge of dredged material does not
 comply with the Guidelines does net
 mean that it can  never be permitted. The
 Act recognizes the concerns of ports in
 section 4O4(bl(2). not 404(b)(l). Many
 readers apparently misunderstood this
 point.
  EPA's role under section 404 is
 several-fold. First, EPA has the
 responsibility for developing the
404(b)(l)  Guidelines in conjunction with
 the Corps. Second. EPA reviews permit
applications and gives its comments (if
any) to the permitting authority. The
Corps may issue a permit even if EPA
comments adversely, after consultation
 takes place.  In the case of state
programs, the State director may not
issue a permit over EPA's unresolved
objection. Third. EPA has the
responsibility for approving and
overseeing State 404  programs. In
addition. EPA has enforcement
responsibilities under section 309.
Finally, under either the Federal or State
program, the Administrator may also
prohibit the specification of a discharge
site, or restrict its use, by following the
procedures set out in section 404(c), if he
determines that discharge would have
an unacceptable adverse effect on fish
and shellfish areas (including spawning
and breeding areas), municipal water
supplies, wildlife or recreation areas. He
may do so in advance of a planned
discharge or while a permit application
is being evaluated or even, in unusual
circumstances, after issuance of a
permit. (See preamble to 40 CFR Part
231. 44 fR 58076. October 9. 1979.) If the
Administrator uses 404(c), he may block
the issuance of a permit by the Corps or
a State 404 program. Where the
Administrator has exercised  his section
4C4(c) authority to prohibit, withhold, or
restrict the specification of a  site for
disposal, his action may not be
overridden under section 404(b)(2). The
fact that EPA has 404(c) authority does
not lessen EPA's responsibility for
developing the 404(b)(l) Guidelines for
use by the permitting authority. Indeed.
if the Guidelines are properly applied.
EPA will rarely have to use its 404(c)
veto.
  The Clean Water Act provides for
several uses of the Guidelines in
addition to the individual permit
application review process described
above. For example, the Corps or an
approved state may issue General
permits  for a category of similar
activities where it determines, on the
b;isis of the 404(b)(V) Guidelines, that
the  activities will cause only  minimal
adverse environmental effects both
individually and cumulatively (Section
404(e) and (g)(l)). In addition, some of
the  exemptions from the permit
requirements involve application of the
Guidelines. Section 404(r) exempts
discharges associated with Federal
construction projects where,  among
other things, there is an Environmental
Impact Statement which considers the
404(b)(l) Guidelines. Section  404(f)(l)(F)
exempts discharges covered by best
management practices (BMP"s)
approved under section 208(b)(4)(B) and
(c),  the approval of which is based in
part on consistency with the  404(b)(l)
Guidelines.
  Several commenters asked for a
statement on the applicability of the
Guidelines to enforcement procedures.
Under sections 309,404(h)(l)(G), and
404(s). EPA. approved States, and the
Corps all play a role in enforcing the
section 404 permit requirements.
Enforcement actions are appropriate
when someone is discharging dredged or
fill material without a required permit
or violates the terms and conditions of a
permit. The Guidelines as such are
generally irrelevant to a determination
of either kind of violation, although they
may represent the basis for particular
permit conditions which are violated.
Under the Corps' procedural regulations.
the Corps may accept an application for
an after-the-fact permit, in lieu of
immediately commencing an
enforcement action. Such after-the-fact
permits may be issued only if they
comply with the 404(b)fl) Guidelines as
well as other requirements set out ;n ihe
Corps' regulations.  Criteria and
procedures for exercising the various
enforcement options are outside the
scope of the section 404(b)(l)
Guidelines.
  Some commenters suggested that we
either include specific permit processing
procedures or that we cross-reference
regulations containing them. Such
procedures are described in 33 CFR Part
320-327 (Corps' procedures) and in 40
CFR Part 122-124 (minimum State
procedures). When specific State 4O4
programs are approved, their regulations
should also be consulted.
How Future Changes In the Testing
Provision Relate to Promulgation of This
Final Rule
  The September 18,1979, proposal
contained testing provisions which -Aere
essentially the same as those in the
Interim Final regulations. The Preamble
to that proposal explained that it was
our intention to propose changes in the
testing provisions, but that a proposal
was not yet ready.  Consequently, while
we  have been revising the rest of the
Guidelines, we have also been working
on a proposal for reorganizing and
updating the testing provisions. Now
that we have finalized the rest of the
Guidelines, two options are available  to
us. First, we could delay issuing any
final revisions to our 1979 proposal until
we  could propose a revised testing
package, consider comments on it. and
finalize the testing  provisions. We could
then put together the Guidelines and the
revised testing section in one final
regulation. The 1975 interim final
Guidelines would apply in their entirety
until then. Second, we could publish the
final Guidelines (with the 1975 testing
provisions) and simultaneously propose
changes to the testing provision. It is our
present belief that  proposed changes to
the testing provision would not affect
the rest of the Guidelines, but the public
would be allowed to comment on any
inconsistencies it saw between the rest
of the Guidelines and the testing
proposal. Then, when the comments to
the testing proposal had been
considered, we would issue a new final
regulation incorporating both the
previously promulgated final Guide!.ne
and the final revised testing provision

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 8S338  Federal Register / Vol. 45. No.  249 / Wednesday.  December  24. 1980 / Rules and Regulations
   We have selected the second option
 Because this approach ensures that.
 needed improvements to the Guidelines
 are made effective at the earliest
 possible date, it gives the public ample
 opportunity to comment on the revised
 testing section, and it maintains the 1975
 testing requirements in effect during the
 interim which would be the case in any
 event.
 Guid line Organization
   M-ny readers objected to the length
 and complexity of the Guidelines. We
 have substantially reorganized the
 regulation to eliminate duplicative
 material and to provide a more logical
 sequence. These changes should make it
 easier for applicants to understand the
 criteria and for State and Corps permit
 evaluators and the Administrator to
 apply the criteria. Throughout the
 document, we have also made numerous
 minor language changes to improve the
 clarity of the regulations, often at  the
 suggestion of commenters.
   Following general introductory
 material and the actual compliance
 requirements, the regulations  are now
 orgarir.ed to more closely follow the
 steps the permitting authority will take
 in arriving at his ultimate decision on
 compliance  with She Guidelines.
   By reorganizing the Guidelines in this
 asriion. we  were also able to  identify
 'and eliminate duplicative material. For
 example, the proposed Guidelines listed
 ways to minimize impacts in many
 separate sections. Since there .was
 substantial overlap in the specific
 methods suggested in those sections, v\e
 consolidated them into new Subpart H.
 Other individual sections have been
 made more concise. In addition, we
 have decreased the number of
 comments, moving them to the Preamble
 or making them part of the Regulation,
 as appropriate.
 General Permits
  When issued after proper
 consideration of the Guidelines, General
 permits arc a useful tool in protecting
 the envirorjmnt  with a minimum of red
 tapp and delay. We expect that their use
 will expand  in the future.
  Some commenters were confused
 about how General permits work.  A
 General permit will be issued  only after
the permitting authority has applied  the
 Guidelines to the class of discharges to
 be covered by the permit. Therefore.
 there is no need to repeat the process al
 the time a particular discharge covered
 by the permit takes place. Of course.
 under both the Corps' regulations and
 EPA's regulations for State programs.
 the permitting authority may suspend
 General permits or require individual
 permits where environmental concerns
 make it appropriate. For example.
 cumulative impacts may turn out to be
 more serious than predicted. This
 regulation is not intended to establish
 the procedures for issuance of General
 permits. That is the responsibility of the
 permitting authority in accordance with
 the requirements of section 404.
 Burden of Proof
  A number of commenters objected to
 the presumption in the regulations in
 general, and in proposed § 230.l(c) in
 particular, that dredged or fill material
 should not be discharged unless it is
 demonstrated that the planned
 discharge meets the Guidelines. These
 commenters thought that it was unfair
 and  inconsistent with section 404(c) of
 the Act.
  We disagree with these objections.
 and  have retained the presumption
 against discharge and the existing
 burden of proof. However, the section
 has been rewritten for clarity.
  The.Clean Water Act itself declares a
 national goal to be the elimination of the
 discharge of pollutants into the
 navigable waters (section 101(a)(l)).
 This goal is  implemented by section 301,
 which states that such discharges are
 unlawful except in compliance  with.
 inter alia, section 404. Section 404 in
 turn authorizes the permitting authority
 to allow discharges of dredged or fill
 material if they comply with the
 404(b)(l) Guidelines. The statutory
 scheme makes it clear that discharges
 shall not take place until they have been
 found acceptable. Of course, this finding
 may be made through the General
 permit process and the statutory
 exemptions  as well as through
 individual permits.
  The commenters who argued that
 section 404(c) shifts the usual burden to
 the EPA Administrator misunderstood
 the relationship between section 404(c)
 and  the permitting process. The
Administrator's authority to prohibit or
restrict a site under section 404(c)
operates independently of  the Secretary
of the Army's permitting authority in
404(a). The Administrator may  use
404(c) whether or not a permit
 application is pending. Conversely, the
 Secretary may deny a permit on the
 basis of the  Guidelines, whether or not
 EPA initiates a 404{c) proceeding. If the
Administrator uses his 404(c) "veto,"
 then he does have the burden to justify
 his action, but that burden does not
come into play until he begins a 404(c)
 proceeding (See 40 CFR Part 231).

Toxic Pollutants
  Many commenters  objected
 strenuously  to the presumptions in the
Guidelines that toxic pollutants on the
section 307(a)(l) list are present in the
aquatic environment unless
demonstrated not to be. and that such
pollutants are biologically available
unless demonstrated otherwise. These
commenters argned that rebutting these
presumptions could involve individual
testing for dozens of substances every
time a discharge is proposed, imposing
an onerous task.
  The proposed regulation attempted to
avoid unnecessary testing by providing
that when the § 230.22(b) "reason to
believe" process indicated that toxics
were not present in the discharge
material, no testing was required. On
the other hand, contaminants other than
toxics required testing if that same
"reason to believe" process indicated
they might be present in the discharge
material. This is in fact a distinction
without a difference. In practical
application, toxic  and non-toxic
contaminants are  treated the same: if
either may be there, tests are performed
to get the information for the
determinations: if it is believed they are
not present, no testing is done. Because
the additional  presumption for toxics
did not actually *°rve a purpose, and
because it was a possible source of
confusion, we  have eliminated it. and
now treat "toxics" and other
contaminants alike, under the "reason to
believe test" (§ 230.60). We have
provided in  § 230.3 a definition of
"contaminants" which encompdsses the
307(a")(l) toxics.

Water Dependency
  One of the provisions in the proposed
Guidelines which received the most
objections was the so-called "water
dependency test" in the proposed
§ 230.10(e). This provision imposed an
additional requirement on fills in
wetlands associated with non-water
dependent activities, namely a  showing
that the activity was "necessary."  Many
environmentalists objected to what they
saw as a substantial weakening of the
1975 version of the water dependency
test. Industry and development-oriented
groups, on the other hand, objected to
the "necessary" requirement because it
was too subjective, and to the provision
as a whole to the  extent that it  seemed
designed to block discharges in
wetlands automatically.
  We have reviewed the water
dependency test  its original purpose.
and its relationship to the rest of the
Guidelines in light of these comments.
The original purpose, which many
commenters commended, was to
recognize the special values of wetlands
and to avoid their unnecessary
destruction, particularly when

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         Federal Register  /  Vol. 45.  No. 249  /  Wednesday. December  24.  1980 / Rules and Regulations  85339
 practicable alternatives were available
 in non-aquatic areas to achieve the
 basic purposes of the proposal. We still
 support this goal, but we have  changed
 the water-dependency test to better
 achieve it.
  First, we agree with the comments
 from both sides that the '"necessary" test
 unpcsed by 'he 1S79 proposal is not
 i'kely to be workable in practice, and
 may spawn more disputes than it settles.
 However, if the "necessary" test is
 simply deleted, section 230.tO(e) does
 not provide any special recognition of or
 protection for wetlands, and thus
 defeats its purpose. Furthermore, even if
 the "necessary" test were retained, the
 provision applies only to discharges of
 f-.il material, not discharges  of dredged
 material, a distinction which lessens the
 effectiveness of the provision. Thus, we
 have decided, in accordance with the
 comments, that the proposal is
 unsatisfactory.
  We have therefore decided to focus
 on. round out. and strengthen the
 approach of the so-called "wntsr
 dependency" provision of the 1975
 reeuUtion. We have rejected tha
 suggestion that we simply go back to the
 1975 language, in part because  it would
 n it mesh easily with the revised general
 provisions of the Guidelines. Instead.
our revised "water de-pendency"
provision creates a presumption that
 Jnere are practicable alternatives to
 "ndn-wa'er dependent" discharges
proposed for speci.il aquatic sites. "Non-
 •.vj'.-r dependent" discharges are those
associated wi:h activities which do not
 require access or proximity  to or siting
 >v:'.h;n the special aquatic site to fulfill
 their basic purpose. An example is a fill
 to create a restaurant site, since
restaurants do not need to be in
wetlands to fulfill their  basic purpose of
feeding people. In the case of such
activities, it is reasonable to assume
there will generally be a practicable site
available upland or in a less vulnerable
part of the aquatic ecosystem. The mere
 fact that an alternative  may cost
 somewhat more does not necessarily
 mean it  is not practicable (see
 § 230.10(a)(2) and discussion below).
Because the applicant may rebut the
 presumption through a clear showing in
 a given case, no unreasonable hardship
should be worked.  At the same time,
 this presumption should have the effect
 of forcing a hard look at the feasibility
of using environmentally preferable
sites. This presumption responds to the
 overwhelming number of commentera
 who urged us to retain a water
 dependency test to discourage
 avoidable discharges in wetlands.
  In addition, the 1975 provision
effectively created a special.
irrebuttable presumption that
alternatives to wetlands were always
less damaging to the aquatic ecosystem.
Because our experience and the
comments indicate that this is not
always the case, and because there
could be substantial impacts on other
fVrrents of the environment and  only
minor impacts on wetlands, we have
chosen instead to impose an explicit, but
rebuttable, presumption that
alternatives to discharges in special
aquatic sites are less damaging to the
aquatic ecosystem and are
environmentally preferable. Of course,
the general requirement that impacts on
the aquatic ecosystem not be
unacceptable also applies. The
legislative history of the Clean Water
Act. Executive Order 11990, and a large
body of scientific information support
th's presumption.
  Apart from the fact that it may  be
rebutted, this second presumption
reir.corporates the key elements of the
19."5 provision. Moreover, it strengthens
it because the recognition of the special
environmental role of wetlands now
applies to all discharges  in special
aquatic sites,  whether of dredged or fill
ma!erial. and whether or not water
dependent. At the same time, this
presumption, like the first cne described
above, retains sufficient flexibility to
reflect the circumstances of unusual
cases.
  Consistent with the general burden of
proof under these Guidelines, where an
applicant proposes to discharge in a
special aquatic site it is his
responsibility to persuade the permitting
authority that both of these
presumptions have clearly been rebutted
in order to pass the alternatives portion
of these Guidelines.
  Therefore, we believe that the new
§ 230.10(a)(3), which replaces proposed
230.10(e). will give special protection to
wetlands and other special  aquatic sites
regardless of material discharged, allay
industry's concerns about the
"necessary" test, recognize the
possibility of impacts on air and upland
sv stems, and acknowledge the
variability among aquatic sites and
discharge activities.

Alternatives
  Some commenters objected at length
to the scope of alternatives which the
Guidelines require to be considered, and
to the requirement that a permit be
denied unless the least harmful such
alternative were selected. Others wrote
to urge us to retain these requirements.
In our judgment, a number of the
objections were based on a
misunderstanding of what the proposed
alternatives analysis required.
Therefore, we have decided to clarify
the regulation, but have not changed its
basic thrust.
  Section 403(c) clearly requires that
alternatives be considered, and provides
the basic legal basis for our requirement.
While the statutory provision leaves the
Agency some discretion to decide how
alternatives are to be considered, we
believe  that the policies and goals of the
Act, as well as the other authorities
cited in the Preamble to the proposed
Guidelines, would be best served by the
approach we  have taken.
  First,  we  emphasize that the only
alternatives which must be considered
are practicable alternatives. What is
practicable depends on cost, technical.
and logistic factors. We have changed
the word "economic" to "cost". Our
intent is to  consider those alternatives
which are reasonable in terms of the
overall scope/cost of the proposed
project. The term economic might be
construed to include consideration of
the applicant's financial standing, or
investment, or market share, a
cumbersome  inquiry which is not
necessarily material to the objectives of
the Guidelines. We consider it implicit
that, to be practicable, an alternative
must be capable of achieving the basic
purpose of  the proposed activity.
Nonetheless,  we have made this explicit
to allay widespread concern. Both
"internal" and "external" alternatives.
as described  in the September 18.1979
Preamble, must satisfy the practicable
test. In order  for an "external"
alternative to be practicable, it must be
reasonably available or obtainable.
However, the mere fact of ownership or
lack thereof, does not necessarily
determine reasonable availability. Some
readers were apparently confused by
the Preamble to the Proposed
Regulation, which referred'to the fact
the National Environmental Policy Act
(NEPA) may require consideration of
courses of action beyond the authority
of the agency involved. We did not
mean to suggest that the Guidelines
were necessarily imposing such a
requirement on private individuals but,
rather, to suggest that what we were
requiring was well within the
alternatives analyses required by NEPA.
   Second, once these practicable
alternatives have been identified in this
fashion, the permitting authority should
consider whether any of them, including
land disposal options, are less
environmentally harmful than the
proposed discharge project Of course,
where there is no significant or easily
identifiable difference in impact, the

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  85340  Federal Register  /  Vol. 45.  No. 249  /  Wednesday. December 24.  I960 / Rules and Regulations
   I tentative need not be considered to
  have "less adverse" impact
   Several commenters questioned the
  legal basis for requiring the permitting
  authority to select the least damaging
  alternative.  (The use of the term "select"
  may have been misleading. Strictly
  speaking, the permitting authority does
  not select anything; he denies the permit
  if the guidelines requirements have not
  been complied with.) As mentioned
  above, the statute leaves to EPA's
  discretion the exact implementation of
  the alternative requirement in section
  403 of the Act. In large part, the
  approach taken by these regulations is
  very similar to that taken by the recent
  section 403(c) regulations (45 FR 65942.
  October 3.1980). There is one difference;
  the Guidelines always prohibit
  discharges where there is a practicable,
  less damaging alternative, while the
  section 403(c) regulations only apply this
  prohibition in some cases. This
  difference reflects the wide range of
  water systems subject to 404 and the
 extreme sensitivity of many of them to
 physical destruction. These waters form
 a priceless mosaic. Thus, if destruction
 of an  area of waters of the United States
 may reasonably be avoided, it should be
 avoided. Of course, where a category of
  04  discharges is so minimal in its
  ffects that it has been placed under a
 general permit, there is no need to
 perform a case-by-case alternatives
 analysis. This feature corresponds, in a
 sense, to the  category of discharges
 under section 403 for which no
 alternatives analysis is required.
   Third, some commenters  were
 concerned that the alternative
 consideration was unduly focused  on
 water quality, and that a better
 alternative from a water quality
 standpoint might be less desirable  from,
 say. an air quality point of view. This
 concern overlooks the explicit provision
 that the existence of an alternative
 which is less damaging to the aquatic
 ecosystem does not disqualify a
 discharge if that alternative has other
 significant advene environmental
 consequences. This laiit provision gives
 the permitting authority an opportunity
 to take into account evidence of damage
 to other ecosystems in deciding whether
 there is a "better" alternative.
  Fourth, a number of commenters were
 concerned that the Guidelines ensure
 coordination  with planning processes
 under the Coastal Zone Management
 Act.  J  208 of the CWA. and other
 programs. We agree that where an
 idequate alternatives analysis has
   ady been  developed, it would be
  asteful not to incorporate it into the
404 process. New 1230.10(a){5) makes it
  clear that where alternatives have been
  reviewed under another process, the
  permitting authority shall consider such
  analysis. However, if the prior analysis
  is not as complete as the alternatives
  analysis required under the Guidelines.
  he must supplement it as needed to
  determine whether the proposed
  discharge complies with the Guidelines.
  Section 230.10(a)(4) recognizes that the
  range of alternatives considered in
  NEPA documents will be sufficient for
  section 404 purposes, where the Corps is
  the permitting authority. (However, a
  greater level of detail may be needed in
  particular cases to be adequate for the
  404(b)(l) Guidelines analysis.) This
  distinction between the Corps and State
  permitting authorities is based on the
  fact that it is the Corps' policy, in
  carrying out its own NEPA
  responsibilities, to supplement ( or
  require a supplement to) a lead agency's
  environmental assessment or impact
  statement where such document docs
  not contain sufficient information. State
  permitting agencies, on the other hand.
  are not subject to NEPA in this manner.
   We have moved proposed
  § 230.10(a)(l) (iii). concerning "other
  particular volumes and concentrations
  of pollutants at other specific rates",
  from the list of alternatives in § 230.10 to
 Subpart H. Minimizing Adverse Effects.
 because it more properly belongs there.

 Definitions (§ 230.3)
   A number of the terms defined in
 § 230.3 are also defined in (he Corps'
 regulation's at 33 CFR 323.2. applicable
 to the Corps' regulatory program. The
 Corps has recently proposed some
 revisions to those regulations and
 expects to receive comments on the
 definitions. To ensure coordination of
 these two sets of regulations, we have
' decided to reserve the definitions of
 "discharge of dredged material."
 "discharge of fill material." "dredged
 material." and "fill material." which
 otherwise would have appeared at
 i 230.3 (f). (g). (j). and (1).
   Although the term "waters of the
 United States" also appears in the
 Corps' regulations, we have retained a
 definition here, in view of the
 importance of this key jurisdictional
 term and the numerous comments
 received. The definition and the
 comments are explained below.
   Until new definitions are published.
 directly or by reference to the Corps'
 revised regulations, users of these
 Guidelines should refer to the
 definitions in 33 CFR 323 J (except in the
 case of state 404 programs, to which the
 definitions in 40 CFR 1122.3 apply.)
   Waters of the United States: A
 number of commenters objected to the
 definition of "waters of the United
 States" because it was allegedly outside
 the scope of the Clean Water Act or of
 the Constitution or because it was not
 identical to the Corps' definition. We
 have retained the proposed definition
 with a few minor changes for clarity for
 several reasons. First a number of
 courts have held that  this basic
 definition of waters of the United States
 reasonably implements section 502(7) of
 the Clean Water Act.  and that it is
 constitutional (e.g.. United States v.
 Byrd. 6O9 F.2d 1204. 7th dr. 1979: Leslie
 Salt Company v. Froehlke. 578 F.2d 742.
 9th Cir. 1978). Second, we agree that it is
 preferable to have a uniform definition
 for waters of the United States, and for
 all regulations and programs under the
 CWA. We have decided to  use the
 wording in the recent  Consolidated
 Permit Regulations. 45 Fed. Reg. 33290.
 May 19.1980. as the standard.*
   Some commenters suggested that the
 reference in the definition to waters
 from which fish are taken to be sold m
 interstate commerce be expanded to
 include areas where such fish spawn.
 While we have not made this change
 because we wish to maintain
 consistency with the wording of the
 Consolidated Permit regulations, we do
 not intend to suggest that a spawning
 area may not have significance for
 commerce. The portion of the definition
 at issue lists major examples, not all the
 ways which commerce may be involved.
   Some reviewers questioned the
 statement in proposed { 230.72(c) (now
 § 230.11(h)) that activities on fast land
 created by a discharge of dredged or fill
 material are considered to be in waters
 of the United States for purposes of
 these Guidelines. The proposed
 language was misleading and we have
 changed it to more accurately reflect our
 intent. When a portion of the Waters of
 the United States has  been legally
 converted to fast land by a discharge of
 dredged or fill material it does not
 remain waters of the United States
 subject to section 301(a).  The discharge
 may be legal because  it was authorized
 by a permit or because it was made
before there was a permit requirement.
 In the case of an illegal discharge, the
 fast land may remain subject to the
jurisdiction of the Act until the
government determines not to seek
restoration. However, in authorizing a
  • The Consolidated Ptnnlt RafulaUont txcludt
certain wait* trtatawnt lyttemi from wattn of ih«
United Suit*. Tht axact terns of thli txclution «r.
unduipini technical raviitont and an txpacted 10
chaos* ihortly. For this nuea tbtM Guidalinei ••
publidMd do not oralaia tbt txcluiioa u orijm.llv
worded ta tht Couolidated Pttmlt Regulation*.
Whta publidMd. tba comcted txchuUm will apply
to OM CofakUaM M w«U u (fa* Couolidated Ptrm.i
RanuUUoat.

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         Federal Register  /  Vol. 45. No. 249  / Wednesday. December  24. 1980 / Rules and Regulations  85341
 discharge which will create fast lands.
 the permitting authority should consider.
 in addition to the direct effects of the fill
 mself, the effects on the aquatic
 environment of any reasonably
 foreseeable activities to be conducted
 on (hut fast land
   Section 230.51 (proposed 230.41) deals
 with impacts on parks, national and
 historical monuments, national sea
 shores, wilderness areas, research sites.
 dad similar preserves. Some readers
 were concerned that we intended the
 Guidelines to apply to activities in such
 preserves whether or not  the activities
 took place in waters of the United
 States. We intended, and we think the
 context makes it clear, that the
 Guidelines apply only to the
 specification of discharge sites in the
 waters of the United States, as defined
 in § 230.3. We have included this section
 because the fact that a water of the
 United States may be located in one of
 these preserves is significant in
 evaluating the  impacts of a discharge
 into that water.
  Wetlands: Many wetlands are waters
 of the United States under the Clean
 Water Act. Wetlands are  also the
 subject of Federal Executive Order No.
 11900. and various Federal and State
 laws and regulations. A number of these
 other programs and laws have
 developed slightly different wetlands
 definitions, in part to accommodate or
 emphasize specialized needs. Some of
 these definitions .nclude.  not only
 wetlands as these Guidelines define
 them, but also mud flats and vegetated
 and unvegetated shallows. Under the
 Guidelines some of these  other'areas are
grouped with wetlands as "Special
Aquatic Sites" (Subpart E) and as such
 their values are given special
 recognition. (See discussion of Water
 Dependency above.) We agree with the
 comment that the National Inventory of
 Wetlands prepared by the U.S. Fish and
Wildlife Service, while not necessarily
 exactly coinciding with the scope of
 waters of the United States under the
Clean Water Act or wetlands under
 these regulations, may help avoid
 construction in wetlands, and be a
 useful long-term planning tool
  Various commenters objected to the
 definition of wetlands in the Guidelines
 as too broad or too vague. This
 proposed definition has been upheld by
 the courts as reasonable and consistent
 with the Clean Water Act and is being
 retained in the final regulation.
 I low ever, we do agree  that \egetative
xuides and other background matenal
 m.iy  be helpful in applying the definition
 in the field. FJ>A and the Corps are
 pledged to work on joint research to aid
in jurisdictional determinations. As we
develop such materials, we will make
them available to the public.
  Other commenters suggested that we
expand the list of examples in the
second sentence of the wetland
definition. While their suggested
additions could legally be added, we
have not done so. The list is one of
examples only, and does not serve as a
limitation on the basic definition. We
are reluctant to start expanding the list.
since there are many kinds of wetlands
which could be included, and the list
could become very unwieldy.
  In addition, we wish to avoid the
confusion which could result from listing
as examples, not only areas which
generally fit the wetland definitions, but
also areas which may or not meet the
definition depending on the particular
circumstances of a given site. In sum. if
an area meets the definition, it is a
wetland for purposes of the Clean Water
Act, whether or not it falls into one of
the listed examples. Of course, more
often than not, it will be one of the listed
examples.
  A few commenters cited alleged
inconsistencies between the definition
of wetlands in § 230.3 and J 230.42.
While we see no inconsistency, we have
shortened the latter section as part of
our effort to eliminate unnecessary
comments.
  Unvegetated Shallows: One of the
special aquatic areas listed in the
proposal was "unvegetated shallows"
(S 230.44). Since special aquatic areas
are subject to the presumptions in
§ 230.10(a)(3), it is important that they
be clearly defined so that the permitting
authority may readily know when to
apply the presumptions. We were
unable to develop, at this time, a
definition for unvegetated shallows
which was both easy to apply and not
too inclusive or exclusive. Therefore, we
have decided the wiser course is to
delete unvegetated shallows from the
special aquatic area classification. Of
course, as waters of the United States,
they are still subject to the rest of the
Guidelines.
  "Fill Material": We are temporarily
reserving J 230.3(1). Both the proposed
Guidelines and the proposed
Consolidated Permit Regulations
defined fill material as material
discharged for the primary purpose of
replacing an aquatic area with dryland
or of changing the bottom elevation of a
water body, reserving to the NPDES
program discharges with the same effect
which  are primarily for the purpose of
disposing of waste. Both proposals
solicited comments on this distinction.
referred to as the primary purpose test.
On May 19.1980, acting  under a court-
imposed deadline. EPA issued final
Consolidated Permit Regulations while
the 404(b)(l) Guidelines rulemaking was
still pending. These Consolidated Permit
Regulations contained a new definition
of fill material which eliminated the
primary purpose test and included as fill
material all pollutants which have the
effect of fill,  that is, which replace part
of the waters of the United States with
dryland or which change the bottom
elevation of a water body for any
purpose. This new definition is similar
to the one used before 1977.
  During the section 404(b)(l)
rulemaking. the  Corps has raised certain
questions about the implementation of
such a definition. Because of the
importance of making the Final
Guidelines available without further
delay, and because of our desire to
cooperate with the Corps in resolving
their concerns about fill material, we
have decided to temporarily reserve
§ 230.3(1) pending further discussion.
This action does not affect the
effectiveness of the Consolidated Permit
Regulations. Consequently, there is a
discrepency between those regulations
and the Corps' regulations, which still
contain the old  definition.
  Therefore, to  avoid any uncertainty
from this situation. EPA wishes to make
clear its enforcement policy for
unpermitted discharges of solid waste.
EPA has authority under section 309 of
the CWA to issue administrative orders
against violations of section 301.
Unpermitted discharges of solid waste
into waters of the United States violate
section 301.
  Under the present circumstances. EPA
plans to issue solid waste administrative
orders with two basic elements. First
the orders will require the violator to
apply to the Corps of Engineers for a
section 404 permit within a specified
period of time. (The Corps has agreed to
accept these applications and to hold
them until it resolves its position on the
definition of fill material.)
  Second, the order will constrain
further  discharges by the  violator. In
extreme cases,  an order may require
that discharges cease immediately.
However, because we recognize that
there will be a lapse of time before
decisions are made on this kind of
permit application, these orders may
expressly allow unpermitted discharges
to continue subject to specific conditions
set forth by EPA in the order. These
conditions will be designed to avoid
further environmental damage.
  Of course, these orders will not
influence the ultimate issuance or non-
issuance of a permit or determine the
conditions that may be specified in sue
a permit. Nor will such orders limit the

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 85342  Federal Register / Vol. 45. No. 249 / Wednesday. December 24. 1980  / Rules  and Regulations
 (Administrator's authority under section
 309(b) or the right of a citizen to bring
 suit against a violator under section 505
 of the CWA.
   Permitting Authority: We have used
 the new term "permitting authority,"
 instead of "District Engineer."
 throughout these regulations, in
 recognition of the fact that under the
 1977 amendments approved States may
 also  issue permits.
 Coastal Zone Management Plans
   Several commenters were concerned
 about the relationship between section
 404 and approved Coastal Zone
 Management (CZM) plans. Some
 expressed concern that the Guidelines
 might authorize a discharge prohibited
 by a  CZM plan: others objected to the
 fact that the Guidelines might prohibit a
 discharge which was consistent with a
 CZM plan.
   Under section 307(b) of the CZM Act
 no Federal permits may be issued until
 the applicant furnishes a certification
 that the discharge is consistent with an
 approved CZM plan, if there is one, and
 the State  concurs in the certification or
 waives review. Section 325.2(b)(2) of the
 Corps' regulation, which applies to all
 Federal 404 permits, implements this
 requirement for section 404. Because  the
 Corps' regulations adequately address
 the CZM consistency requirement, we
 have  not duplicated § 325.2(b)(2) in the
 Guidelines. Where a State issues State
 404 permits, it may of course require
 consistency with  its CZM plan under
 State law.
   The second concern, that the 404
 Guidelines might  be stricter than a CZM
 plan,  points out a possible problem with
 CZM plans, not with the Guidelines.
 Under 307(f) of CZMA. all CZM plans
 must  provide for compliance with
 applicable requirements of the Clean
 Water Act. The Guidelines are one such
 requirement. Of course, to the extent
 that a CZM plan is general and area-
 wide, it may be impossible to include in
 its development the same project-
 specific consideration of impacts and
 alternatives required under the
 Guidelines. Nonetheless, it cannot
 authorize  or mandate a discharge of
 dredged or fill material which fails to
 comply with the requirements of these
 Guidelines. Often CZM plans contain a
 requirement that all activities conducted
 under it meet the permit requirements of
 the Clean Water Act. In such a case,
 there  could of course be no conflict
 between the CZM plan and the
requirements of the Guidelines.
  We agree with commenters who urge
that delay and duplication of effort be
avoided by consolidating alternatives
studies required under different statutes,
 including the Coastal Zone Management
 Act. However, since some planning
 processes do not deal with specific
 projects, their consideration of
 alternatives may not be sufficient for the
 Guidelines. Where another alternative
 analysis is less complete than that
 contemplated under section 404. it may
 not be used to weaken the requirements
 of the Guidelines.
 Advanced Identification of Dredged or
 Fill Material Disposal Sites
   A large number of commenters
 objected to the way proposed § 230.70.
 new Subpart I. had been changed from
 the 1975 regulations. A few objected to
 the section itself. Most of the comments
 also revealed a misunderstanding about
 the significance of identifying an area.
 First, the fact that an area has been
 identified as unsuitable for a potential
 discharge site does not mean that
 someone cannot apply  for and obtain a
 permit to discharge  there as long as the
 Guidelines and other applicable
 requirements are satisified.' Conversely,
 the fact that an area has been identified
 as a potential site does not mean that a
 permit is unnecessary or that one will
 automatically be forthcoming. The intent
 of this section was to aid applicants by
 giving advance notice that they would
 have a relatively easy or difficult time
 qualifying for a permit to use particular
 areas. Such advance notice should
 facilitate applicant planning and shorten
•permit processing time.
   Most of the objectors focused on
 EPA's "abandonment"  of its "authority"
 to identify sites. While that "authority"
 is perhaps less "authoritative" than the
 commenters suggested  (see above), we
 agree that there is no reason to decrease
 EPA's role in the process. Therefore, we
 have changed new § 230.80(a) to read:
  "Consistent with these Cuidelinea. EPA
 and the permitting authority on their own
 initiative or at the request of any other party,
 and after consultation with any affected State
 that is not the permitting authority, may
 identify sites which will be considered as:"
   We have also deleted proposed
 § 230.70(a)(3), because it did not seem to
 accomplish much. Consideration of the
 point at which cumulative and
 secondary impacts become
 unacceptable and warrant emergency
 action will generally be more
 appropriate in a permit-by-permit
 context Once that point has been so
 determined, of course, the area can be
 identified as "unsuitable" under the new
 5 230.80(a)(2).
  • EPA may fondofc the UM of a lilt by
exercising ill authority under lection 404(c). The
•dvince identification referred to In thii lection ii
not • Mction «M(c) prohibition.
Executive Order 12044
  A number of commenters took the
position that Executive Order 12044
requires EPA to prepare a "regulatory
analysis" in connection with these
regulations. EPA disagrees. These
regulations are not. strictly speaking,
new regulations. They do not impose
new standards or requirements, but
rather substantially clarify and
reorganize the existing in'.enrr. final
regulations
  Under EPA's criteria implementing
Executive Order 12044. EPA will prepare
a Regulatory Analysis for any regulation
which imposes additional annual costs
totalling $100 million or which will result
in a total additional cost of production
of any major product or service  which
exceeds 5% of its selling price. While
many commenters. particularly
members of the American Association
of Port Authorities (AAPA), requested a
regulatory analysis and claimed that the
regulations were too burdensome, none
of them explained how that burden was
an additional one attributable to this
revision. A close comparison of the new
regulation and the explicit and implicit
requirements in the interim final
Guidelines reveals that there has been
very little real change  in the  criteria by
which discharges are to be judged or in
the tests that must be conducted:
therefore, we stand by our original
determination that a regulatory  anal; sis
is not required.
  Perhaps the most significant area in
which the regulations are more explicit
and arguably stricter is in the
consideration of alternatives. Howexer.
even the 1975 regulations required the
permitting authority to consider "the
availability of alternate sites and
methods of disposal that are less
damaging to the environment." and (o
avoid activities which would have
significant adverse effects. We do not
think  that the revised Guidelines' more
explicit direction to avoid adverse
effects that could be prevented through
selection of a clearly less damaging site
or method is a change imposing a
substantial new burden on the regulated
public.
  Because the revised regulations are
more explicit than the interim final
regulations in some respects, it is
possible that permit reviewers will do a
more thorough job evaluating proposed
discharges. This may result in somewhat
more carefully drawn permit conditions.
However, even if. for purposes of
argument the possible cost of complying
with these conditions is considered an
additional cost there is no reason to
believe that it alone will be anywhere
near $100 million annually.

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         Federal Register  /  Vol. 45.  No. 249  /  Wednesday. December  24. 1980 / Rules  and  Regulations  85343
   We also believe that it is appropriate
 to recognize the regulatory benefits from
 these more carefully drafted final
 regulations. Because they are much
 clearer about what should be considered
 and documented, we expect.thrre will
 ho fewer delays n re\ iPivinR p.Tnuis
 ..  J th i! mit'H! ^pri« :TS to •>• 'ue
                                   '0
       authority Th"s<. b' nef is r.re
 expected to offset ;ir:y potential cc^t
 inr.rease.
   Some commenters suggested that
 documentation requirements would
 generate an additional cost of
 operations. The Corps' procedural
 regulations at 33 CFR 325.8 ui.d 325.11
 already to quire extensive
 documentation for individual permits
 being denied or being referred to higher
 authority for resolution of a conflict
 between agencies.

 Economic Factors
   A number of commfinters asked EPA
 !o include consideration of economic
 factors in the Guidelines. We believe
 that the regulation  already recognizes
 economic factors to the extent
 contemplated by the statute. First, the
 Guidelines explicitly include the concept
 of "practicability" in connection with
 both altern.itivt-s and steps to minimize
 impar ;s  If an alleged alternative is
 •jnrsasonably expensive to the
 .ippl;cant. the alternative is not
 ' practicjble. ' In addition. Ihe
 C'.aidehnes also consider cc,oncmiLS
 indirectly in lhat they are structured to
 ;.'void the expense of unnecessary
 testing through  the  "reason-to-believe-
 tust." Second, the statute expressly
 provides that the economics of
 anchorage and navigation may be
 considered, but only after application of
 Ihe section 404(b)(l) Guidelines. (See
section 404(b)(2).)
 Borrow Sites
  A number of highway departments
objected because they felt the
Guidelines would require them to
 identify specific borrow sites at the time
of application, which would disrupt their
normal contracting process and increase
cost. These objections were based on a
 misunderstanding of the Guideline's
requirements. Under those Guidelines.
 the actual borrow sites need not be
 identified, if the application and the
permit specify that the discharge
material  must come from clean upland
sites which are removed from sources of
contamination and otherwise satisfy the
reason-to-believe test A condition that
 the material come from such a site
would enable the permitting authority to
 make his determinations and find
compliance with the conditions of
§ 230.10. without requiring highway
departments to specify in advance the
specific borrow sites to be used.
Consultation With Fish and Wildlife
Agencies
  One commenter wanted us to put in a
statement that the Fish and Wildlife
Coordination Act requires consultation
with fish and wildlife agencies. We have
not added new language because (1) the
Fish and Wildlife Act only applies to
Federal permitting agencies and not to
State permitting agencies, and (2) the
Corps' regulations already provide for
such consultation by the only Federal
404 permitting  agency. However, we
agree with the  commenter that Federal
and State  fish and wildlife agencies may
often provide valuable assistance in
evaluating the  impacts of discharges of
dredged or fill  material.
The Importance of Appropriate
Documentation
  Specific documentation is important
to ensure an understanding of the basis
for each decision to allow, condition, or
prohibit a  discharge through application
of the Guidelines. Documentation of
information is  required for (1) facts and
data gathered in the evaluation and
testing of the extraction site, the
material to be discharged, and the
disposal site; (2)  factual determinations
regarding changes that can be expected
at the disposal site if the discharge is
made as proposed; and (3) findings
regarding compliance with § 230.10
conditions. This documentation provides
a record of actions taken that can be
evaluated  for adequacy and accuracy
and ensures consideration of all
important  impacts in the evaluation of a
proposed discharge of dredged or fill
material.
  The specific  information documented
under (1) and (2) above in any given
case depends on the level of
investigation necessary to provide for a
reasonable understanding of the impact
on the aquatic  ecosystems. We
anticipate that a number of individual
and most General permit applications
will be for routine, minor activities with
little potential  for significant adverse
envircnmental impacts. In such cases,
the permitting  authority  will not have to
require extensive testing or analysis to
make his findings of compliance. The
level of documentation should reflect
the significance and complexity of the
proposed discharge activity.

Factual Determinations
  Proposed section 230.20, "Factual
Determinations" (now $ 230.11) has
been significantly reorganized in
response to comments. First we have
changed (e) to reflect our elimination of
the artificial distinction between the
section 307(a)(l) toxics and other
contaminants. Second, we  have
eliminated proposed (f) (Biological
Availabi'ityj, since the necessary
information will be provided by (d) and
new (e). Proposed (f) was intended  to
reflect the presumption that toxics were
present and biologically available. We
have modified proposed (g). now (f). to
focus  on the size of the disposal site and
the size  and shape of the mixing zone.
The specific requirement to document
the site has been deleted; where such
information is relevant, it will
automatically be considered  in making
the other determinations. We have also
deleted proposed (h) (Special
Determinations) since it did not provide
any useful information whi.ch would not
already  be considered in making the
other  factual determinations.
  Finally, in response to many
comments, we have moved the
provisions on cumulative and secondary
impact to the Factual Determination
section to give them further emphasis.
We agree that such impacts are an
important consideration in evaluating
the acceptability of a discharge site.
Water Quality Standards
  One commenter was concerned that
the reference $  230.10(b) to water
quality standards and criteria
"approved or promulgated under section
303" might encourage permit authorities
to ignore other water quality
requirements. Under section 303. all
State  water quality standards are to be
submitted to EPA for approval. If the
submitted standards are incomplete or
insufficiently stringent EPA  may
promulgate standards to replace or
supplant the State standards.
Disapproved standards remain in effect
until replaced. Therefore, to refer to
"EPA approved or promulgated
standards" is to ignore those State
standards which have been neither
approved nor replaced. We have
therefore changed the wording*of this
requirement as follows:	any
applicable State water quality
standard" We  have also dropped the
reference to "criteria", to be consistent
with the Agency's general position that
water quality criteria are not regulatory.
Other Requirements for Discharge
   Section 230.10(c) provides that
discharges are  not permitted if they will
have  "significantly" adverse effects on
various aquatic resources. In this
context "significant" and  "significantly'
mean more than "trivial",  that is.
significant in a conceptual rather than a
statistical sense. Not all effects which

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85344  Federal Register / Vol. 45, No. 249 /  Wednesday, December 24, 1980 / Rules  and  Regulations
  re statistically significant in the
  laboratory are significantly adverse in
 the field.
   Section 320.10(d) uses the term
 "minimize" to indicate that all
 reasonable reduction in impacts be
 obtained. As indicated by the
 "appropriate and practicable" provision.
 steps which would be unreasonably
 costly or would be infeasible or which
 would accomplish only inconsequential
 reductions in impact need not be taken.
 Habitat Development and Restoration of
 Water Bodies
   Habitat development and restoration
 involve changes in open water and
 wetlands that minimize adverse effects
 of proposed changes or that neutralize
 or reverse the effects of past changes on
 the ecosystem. Development may
 produce a new or modified ecological
 state by displacement of some or all of
 the existing environmental
 characteristics. Restoration has the
 potential to return degraded
 environments to their former ecological
 state.
   Habitat development and restoration
 car. contribute to the maintenance  and
 enhancement of a viable aquatic
 ecosystem at the discharge site. From an
 ^environmental point of view, a project
  volving the discharge of dredged and
  11 material should be designed and
 managed to emulate a natural
 ecosystem. Research, demonstration
 projects, and full scale implementation
 have been done in many categories of
 development and restoration. The U.S.
 Fish and Wildlife Service has programs
 to develop and restore habitat. The U.S.
 Army Engineer Waterways Experiment
 Station has published guidelines for
 using dredged material to develop
 wetland habitat, for establishing marsh
 vegetation, and for building islands that
 attract colonies of nesting birds. The
 EPA has a Clean Lakes program which
 supplies funds to States and localities to
 enhance or restore degraded lakes. This
 may involve dredging nutrient-laden
 sediments from a lake and ensuring that
 nutrient inflows  to the lake are
 controlled. Restoration and habitat
 development techniques can be used to
 minimize adverse impacts and
 compensate for destroyed habitat
 Restoration and  habitat development
 may also provide secondary benefits
 such as improved opportunities for .
 outdoor recreation and positive use for
 dredged materials.
  The development and restoration of
 viable habitats in water bodies requires
 fanning and construction practices that
 ' tegrate the new or improved habitat
  to the existing environment. Planning
requires a model or standard, the
                                        achievement of which is attempted by
                                        manipulating design and implementation
                                        of the activity. This model or standard
                                        should be based on characteristics of a
                                        natural ecosystem in the vicinity of a
                                        proposed activity. Such use of a natural
                                        ecosystem ensures that the developed or
                                        restored area, once established, will be
                                        nourished and maintained physically,
                                        chemically and biologically by natural
                                        processes.  Some examples of natural
                                        ecosystems include, but are not limited
                                        to. the following: salt marsh, cattail
                                        marsh, turtle grass bed, small island, etc.
                                         Habitat development and restoration,
                                        by definition, should have
                                        environmental enhancement and
                                        maintenance as their initial purpose.
                                        Human uses may benefit but they are
                                        not the primary purpose. Where such
                                        projects are not founded on the
                                        objectives of maintaining ecosystem
                                        function and integrity, some values may
                                        be favored at the expense of others. The
                                        ecosystem  affected must be considered
                                        in order to  achieve the  desired result of
                                        development and restoration. In the
                                        final analysis, selection of the
                                        ecosystem  to be emulated is of critical
                                        importance and a loss of value can
                                        occur if the wrong model or an
                                        incomplete model is selected. Of equal
                                        importance is the planning and
                                        management of habitat development
                                        and restoration on a case-by-case basis.
                                         Specific measures to minimize
                                        impacts on the aquatic ecosystem by
                                        enhancement and restoration projects
                                        include  but are not limited to:
                                         (t) Selecting the nearest similar
                                        natural ecosystem as the model in the
                                        implementation of the activity.
                                         Obviously degraded or significantly
                                        less productive habitats may be
                                       considered prime candidates for habitat
                                       restoration. One viable habitat,
                                       however should not be sacrificed in an
                                       attempt to create another, i.e.. a
                                       productive  vegetated shallow water
                                       area should not be destroyed in an
                                       attempt to create a wetland in its place.
                                         (2) Using development and restoration
                                       techniques  that have been demonstrated
                                       to be effective in circumstances similar
                                       to those under consideration wherever
                                       possible.
                                         (3) Where development and
                                       restoration techniques proposed for use
                                       have not yet advanced  to the pilot
                                       demonsVration or implementation stage.
                                       initiate their use on a small scale to
                                       allow corrective action if unanticipated
                                       adverse impacts occur.
                                         (4) Where Federal funds are spent to
                                       clean up waters of the U.S. through
                                       dredging, scientifically  defensible levels
                                       of pollutant concentration in the return
                                       discharge should be agreed upon with
                                       the funding authority in addition to any
applicable water quality standards in
order to maintain the desired improved
water quality.
  (5) When a significant ecological
change in the aquatic environment is
proposed by the discharge of dredged or
fill material, the permitting authority
should consider the ecosystem that will
be lost as well as the environmental
benefits of the new system.
  Dated. December 12. 1980.
Douglas M. Cotlle,
Administrator. Environmental Protection
Agency.
  Part 230 is revised to read as follows:

PART 230—SECTION 404(bX1)
QUIOEUNES FOR SPECIFICATION OR
DISPOSAL SITES FOR DREDGED OF
FILL MATERIAL

Subpart A—General

Sec.
230.1  Purpose and policy.
230.2  Applicability.
230.3  Definitions.
230.4  Organization.
230.5  General procedures to be followed.
230.6  Adaptability.
230.7  General permits.

Subpart B—Compliance With the Guidelines
230.10  Restrictions on discharge.
230.11  Factual determinations.
230.12  Findings of compliance or  non-
    compliance with the restrictions on
    discharge.

Subpart C—Potential Impacts on  Physical
and Chemical Characteristics of the
Aquatic Ecosystem
230 20  Substrate.
230.21  Suspended particulates/turbidity
230 22  Water.
230.23  Current patterns and water
    circulation.
230.24  Normal water fluctuations.
230.25  Salinity gradients.

Subpart 0—Potential Impacts on  Biological
Characteristics of the Aquatic Ecosystem
230.30  Threatened and endangered species.
230.31  Fish, crustaceans, mollusks. and
    other aquatic organism* in the food web.
230.32  Other wildlife.

Subpart E—Potential Impacts on Special
Aquatic Sites
       Sanctuaries and refuges.
       Wetlands.
       Mud flats.
       Vegetated shallows.
       Coral reefs.
       Riffle and pool complexes.
230.40
230.41
2J0.42
230.43
230.44
230.45

Subpart F—Potential Effects on Human Use
Characteristic*
230.50  Municipal and private water
    supplies.
230.51  Recreational and commercial
    fisheries.
230.52  Water-related recreation.
230.53  Aesthetics.

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         Federal  Register / Vol.  45.  No. 249  /  Wednesday. December 24, 1980 / Rules  and  Regulations  85345
 Sec.
 230.54 Parks, national and historic
    monuments, national seashores.
    wilderness areas, research sites and
    similar preserves.
 Subpart Q—Evaluation and Testing
 230.60 General evaluation of dredged or fill
    material.
 230 61 Chemical biological and physical
    evaluation and testing.
 Subpart H—Actions to Minimize Adverse
 Effects
 230 "0 Actions concerning the location of
    the discharge.
 23071 Actions concerning the material to be
    discharged.
 23(1.72 Actions controlling the material after
    discharge.
 230.73 Ac'ions affecting the method of
    dispersion.
 230.74 Actions related to technology.
 230.75 Actions affecting plant and animal
    popu'ations.
 230.78 Actions affecting human use.
 230.77 Other actions.
 Subpart I—Planning To Shorten Permit
 Processing Time
 230 80 Advanced identification of disposal
    areas.
  Authority: This regulation is issued under
 authority of Sections 404(b) and 5Cl(a) of the
 Clean Wdter Act of 1977. 33 U S C. § I344(b)
 d.-,d §  1361 (a).

 Subpart A—General
 §23.1  Purpose and policy.
  (a)  The purpose of these Guidelines is
 to restore and maintain the chemical,
 physical, and biological integrity of
 waters of the United States through the
 control of discharges of dredged or fill
 material.
  (b)  Congress has expressed a number
 of policies in the Clean Water Act.
 These Guidelines are intended to be
 consistent with and to implement those
 policies.
  (r) Fundamental to these Guidelines is
 the precept that dredged or Till material
 should not be discharged into the
 aquatic ecosystem, uHess it can be
 demonstrated that sudi a discharge will
 not have an unacceptable adverse
 impact either individually or in
 combination with known and/or
 probable impacts of other activities
 affecting the ecosystem* of concern.
  (d)  From a national perspective, the
 degradation or destruction of special
 aquatic sites, such as filling operations
 in wetlands, is considered to be among
 the most severe environmental impacts
covered by these Guidelines. The
guiding principle should be that
degradation or destruction of special
sites may represent an irreversible loss
of valuable aquatic resources.
{230.2  Applicability.
  (a) These Guidelines have been
developed by the Administrator of the
Environmental Protection Agency in
conjunction with the Secretary of the
Army acting through the Chief of
Engineers under section 404(b)(l) of the
Clean Water Act (33 U.S.C. 1344). The
Guidelines  are applicable to the
specification of disposal sites for
discharges  of dredged or fill material
into waters of the United States. Sites
may be specified through:
  (1) The regulatory program of the U.S.
Army Corps of Engineers under sections
404(a) and (e) of the Act (see 33 CFR
320. 323 and 325);
  (2) The civil works program of the U.S.
Army Corps of Engineers (see 33  CFR
209.145 and section 150, of Pub. L 94-587.
Water Resources Development Act of
1976);
  (3) Permit programs of States
approved by the Administrator of the
Environmental Protection Agency in
accordance with sections 404(g) and (h)
of the Act (see 40 CFR 122.123 and 124);
  (4) Statewide dredged or fill material
regulatory programs with best
management practices  approved  under
section 208(b)(41(B) and (C) of the Act
(see 40 CFR 35.1560);'
  (5) Federal construction projects
which meet criteria specified in section
404(r) of the Act.
  (b) These Guidelines will be applied
in the review of proposed discharges of
dredged or  fill material into navigable
waters which lie inside the baseline
from which the territorial sea  is
measured, and the discharge of fill
material into the territorial sea, pursuant
to the procedures referred to in
paragraphs (a)(l) and (a)(2) above. The
discharge of dredged material into the
territorial sea is governed by the Marine
Protection,  Research, and Sanctuaries
Act of 1972, Pub. L 92-532. and
regulations and criteria issued pursuant
thereto (40  CFR Part 220-228).
  (c) Guidance on  interpreting and
implementing these Guidelines may be
prepared jointly by EPA and the Corps
at the national or regional level from
time to time. No modifications to the
basic application, meaning, or intent of
these Guidelines will be made without
rulemaking by the  Administrator under
the Administrative Procedure Act (5
U.S.C. SSlefse?.).

§2304  DeflnlUone.
  For purposes of this Part, the
following terms shall have the meanings
indicated:
  (a) The term "Act" means the Clean
Water Act  (also known as the Federal
Water Pollution Control Act or FWPCA)
Pub. L. 92-500, as amended by Pub. L.
95-217. 33 U.S.C. 1251. et seq.
  (b) The term "adjacenf'means
bordering, contiguous, or neighboring.
Wetlands separated from other waters
of the United States by man-made dikes
or barriers, natural river berras. beach
dunes, and the like are "adjacent
wetlands."
  (c) The terms "aquatic environment"
and "aquatic ecosystem" mean waters
of the United States, including  wetlands.
that serve as habitat for interrelated and
interacting commumtiss and populations
of plants and animals.
  (d) The term "carrier of contarn.nant"
means dredged or fill material that
contains contaminants.
  (e) The term "contaminant" means a
chemical or biological substance in a
form that can be incorporated into, onto
or be ingested by and that harms
aquatic organisms, consumers of aquatic
organisms, or users of the aquatic
environment, and includes but is not
limited to  the substances on the
307(a)(l) list of toxic pollutants
promulgated on January 31, 1978 (43 FR
4109).
  (f) [Reserved]
  (g) [Reserved)
  (h) The term "discharge point" means
the point within the disposal site at
which the dredged or fill material  is
released.
  (H The term "disposal site" means
that portion of the "waters of the L'nited
States" where specific disposal
activities are permitted and consist of a
bottom surface area and any o\eHying
volume of water. In the case of wetlands
on which surface water is not present.
the disposal site consists of the wetland
surface area.
  (j) [Reserved]
  (k) The term "extraction sito" means
the place from which the dredged or fill
material proposed for discharge is to be
removed.
  (1) [Reserved]
  (m) The term "mixing zone"  means a
limited volume of water serving as a
zone of initial dilution in the immediate
vicinity of a discharge point where
receiving water quality may not meet
quality standards or other requirements
otherwise applicable to the receiving
water. The mixing zone should be
considered as a place where wastes and
water mix and not as a place where
effluents are treated.
  (n) The term "permitting authority"
means the District Engineer of the U.S.
Army Corps of Engineers or such other
individual as may be designated by the
Secretary of the Army to issue or deny
permits under section 404 of the Act or
the State Director of a permit program

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  85346 Federal Register / Vol. 45. No.  249 / Wednesday. December 24. 1930  /  Rules and Regulations
  approved by EPA under } 404(g) and
  S 404(h) or his delegated representative.
   (o) The term "pollutant" means
  dredged spoil, solid waste, incinerator
  residue, sewage, garbage, sewage
  sludge, munitions, chemical wastes,
  biological materials, radioactive
  materials not covered by the A'orr.ic
  Energy Act. heat, wreck r-d or d scarded
  equipment, rock. sand, cellar dirt, and
  industrial, municipal, and agricultural
  waste discharged into water. The
  legislative history of the Act reflects that
  "radioactive materials" as included
 within tr>e definition of "pollutant"  in
 section 502 of the Act means only
 radioactive materials which are not
 encompassed in the definition of source,
 byproduct, or special nuclear materials
 as defined by the Atomic Energy Act of
 1954. as amended, and regulated under
 the Atomic Energy Act. Examples of
 radioactive materials not covered by the
 Atomic Energy Act and, therefore,
 included within the term "pollutant", are
 radium and accelerator produced
 isotopes See Train v Colorado Pub/:c
 '•• Seres- 1 Research Croup. Inc.. 426 U S. 1
 i l'.TG)
   (p) The term "pollution ' moans the
 man-n^d? or man-induced alteration of
 the chemical, physical, biological or
          al integrity of an aquatic
 ecosystem.
   [c,j The term "practicable" means
 a\ ailable and capable of being done
 after taking into consideration cost,
 ex'siinj technology, and logistics in light
 of overall project purposes.
   (q-1) "Special aquatic sites" means
 those sites identified in Subpart E. They
 are geographic areas, large or small.
 possessing special ecological
 characteristics of productivity, habitat,
 wildlife protection, or other important
 and easily disrupted ecological values.
 These areas are generally recognized as
 significantly influencing or positively
 contributing to the general overall
 environmental health or vitality of the
 entire ecosystem of a region. (See
 230.10(a)(3))
  {rj The term "territorial  sea" means
 the belt of the sea measured from the
 baseline as determined in accordance
 with the Conventon on the Territorial
 Sea and the Contiguous Zone and
 extending seaward a distance of three
 miles.
  (s) The term "waters of  the united
 States" means:
  (1) All waters  which are currently
used, or were used in the past, or may
be susceptible to use in interstate or
foreign commerce, including all waters
which are subject to the ebb and flow of
the tide:
  (2) All interstate waters including
interstate  wetlands;
   (3) All other waters auch as intrastate
 lakes, rivers, streams (including
 intermittent streams), mudflats,
 sandflats. wetlands, sloughs, prairie
 potholes, wet meadows, playa lakes, or
 natural ponds, the use, degradation or
 destruction of which could affect
 interstate or foreign commerce including
 any such waters:
   (i) Which are or could be used by
 interstate or foreign travelers for
 recreational or other purposes: or
   (ii) From which fish or shellfish are or
 could be taken and sold in interstate or
 foreign commerce; or
   (lii) Which are used or could be used
 for industrial purposes by industries in
 interstate commerce;
   (4) All impoundments of waters
 otherwise defined as waters of the
 United States under this definition.
   (5) Tributaries of waters identified in
 paragraphs (t)-(4) of this section:
   (6) The territorial sea;
   (7) Wetlands adjacent to waters
 (other than waters that are themselves
 wetlands) identified in paragraphs  (s)
 (l)-(fi) of this section; waste  treatment
 systems, including treatment ponds or
 lagoons designed to meet the
 requirements of CWA (other than
 cooling ponds as defined in 40 CFR
 § 423.ll(m) which also meet  the criteria
 of this definition) are not waters of the
 United States.
   (t) The term "wetlands" means those-.
 areas that are inundated or saturated by
 surface or ground water at a frequency
 and duration  sufficient to support, and
 that under normal circumstances do
 support, a prevalence of vegetation
 typically adapted for life in saturated
 soil conditions. Wetlands generally
 include swamps, marshes, bogs and
 similar areas.

 §230.4 Organization.
  The Guidelines are divided into eight
 subparts. Subpart A presents those
provisions of general applicability,  such
as purpose and definitions. Subpart B
establishes the four conditions which
 must be satisfied in order to  make a
 finding that a proposed discharge of
dredged or fill material complies with
 the Guidelines. Section 230.11 of Subpart
B. sets forth factual determinations
 which are to be considered in
determining whether or not a proposed
discharge satisfies the Subpart B
conditions of compliance. Subpart C
describes the physical and chemical
components of a site and provides
guidance as to how proposed discharges
of dredged or fill material may affect
 these components. Subparts  D-F detail
 the special characteristics of particular
 aquatic ecosystems in terms  of their
 values, and the possible loss of these
values due to discharges of dredged or
fill material. Subpart G prescribes a
number of physical, chemical, and
biological evaluations and testing
procedures to be used in reaching the
required factual determinations. Subpart
H details the means to prevent or
mimimize adverse effects. Subpart  I
concerns advanced identifica'ion r:
disposal areas.

§ 230.5  General procedures to b«
followed.
  In evaluating whether a particular
discharge site may be specified, the
permitting authority should use  these
Guidelines in the following sequence:
  (a) In order to obtain an overs iew of
the principal regulatory provisions  of the
Guidelines, review the restrictions  en
discharge in § 230.10(a)-(d). the -
measures to mimimize adverse inpnct cf
Subpart H, and the required factu-21.
determinations  of § 230.11.
  (b) Determine if a General pern-.:1.
(§ 230.7) is applicable; if so. the
applicant needs merely to corr.piv :\>.:h
:ts terms, and no further artion bv  -ue
permitting authority is necessary
Specia. conditions for evaluat.nn ™f
proposed General permits are c(
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         Federal Register  /  Vol. 45. No. 249 / Wednesday.  December 24.  1980 / Rules and Regulations  85347
   (i) If there is a reasonable probability
 of chemical contamination, conduct the
 appropriate tests according to the
 section on Evaluation and Testing
 l§ 23061).
   !jl Identify appropriate and
 practicable changes to the project plan
 to minimize the environmental impact of
 'he ii:s<. barge, bdsed upon the
 •.;)! i '.ir.zed methods of min.mizjtion of
 ..-rpacts in Subpart H.
   |k) Make and document Factual
 Determinations in § 230.11.
   (i) Make and document Findings of
 Compliance (§ 230.12) by comparing
 Factual Determinations with the
 requirements for discharge of §  230.10.
 This outline of the steps to follow in
 usinq the Guidelines is simplified for
 purposes of illustration. The actual
 process followed may be iterative, with
 the results erf one step leading to a
 reexdmination of previous steps. The
 permitting authority must address all of
 the relevant provisions of the Guidelines
 in rsng a  Finding of Compliance in
 an individual case.

 * 210.6  Adaptability.
   ;.i! TIP -r.anr.rr In '.vhich these
 Guiue'°.!iL-s are usc-d depends on the
 physi'-ai. biological, and chemical nature
 of tht; proposed extraction site,  the
 mdtfvuii to Lu discharged, dnd the
 r. '.r.didaie disposal site, including any
 other important components of the
 c.',.j<:>s'em !ierng evaluated.
 Documentation to demonstrate
 knouiedge about the extraction site.
 aidteridls to be extracted, and the
 candidate disposal site is an essential
component of guideline application.
These Guidelines allow evaluation and
documentation for a variety of activities,
 ranging from those with large, complex
 impacts on the aquatic environment to
 those for which the impact is likely to be
 innocuous. It  is unlikely that the
 Guidelines will apply in their entirety to
 any one activity, no matter how
 complex. It is anticipated that
 substantial numbers of permit
 applications will be for minor, routine
 activities that have little, if any.
 potential for significant degradation of
 the aquatic environment It generally is
 not intended or expected that extensive
 testing, evaluation or analysis will be
 needed  to make findings of compliance
 in such routine cases. Where the
conditions for General permits are met,
and where numerous applications for
 similar activities are likely, the  use of
 General permits will eliminate repetitive
 evaluation and documentation for
 individual discharges.
   (b) The Guidelines user, including the
 agency or agencies responsible  for
implementing the Guidelines, must
recognize the different levels of effort
that should be associated with varying
degrees of impact and require or prepare
commensurate documentation. The level
of documentation should reflect the
significance and complexity of the
discharge activity.
  (c) An essential part of the evaluation
process involves making determinations
as to the relevance of any portion(s) of
the  Guidelines and conducting further
evaluation only as needed. However,
where portions of the Guidelines review
procedure are "short form" evaluations,
there still must be sufficient information
(including consideration of both
individual and cumulative impacts) to
support the decision of whether to
specify the site for disposal of dredged
or fill material and  to support the
decision to curtail or abbreviate the
evaluation process. The presumption
against the discharge in J 230.1 applies
to this decision-making.
  (d) In the case of activities covered by
General permits or 208(b)(4)(B) and (C)
Best Management Practices, the analysis
and documentation required by the
Ciuuelir.es will be performed at the time
of General permit issuance or
208(b)(4)iB) and (C) Best Management
Practices promulgation and will not be
repeated when activities are conducted
under a General permit or 208(b)(4)(B)
and (C) Best Management Practices
control. These Guidelines do not require
reporting or formal written
communication at the time individual
activities are initiated under a General
permit or 208(b)(4)(B) and (C) Best
Management Practices. However, a
particular General permit may require
appropriate reporting.
5 230.7  G«n«ral p«rmlt«.
  (a) Conditions for the issuance of
General permits. A General permit for a
category of activities involving the
discharge of dredged or fill material
complies with the Guidelines if it meets
the  applicable restrictions on the
discharge in $ 230.10 and if the
permitting authority determines that:
  (1) The activities in such category are
similar in nature and similar in their
impact upon water quality and the
aquatic environment;
  (2) The activities in such category will
have only minimal adverse effects when
performed separately; and
  (3) The activities in such category will
have only minimal cumulative adverse
effects on water quality and the aquatic
environment.
  (b) Evaluation process. To reach the
determinations required in paragraph (a)
of this section, the permitting authority
shall set forth in writing an evaluation of
the potential individual and cumulative
impacts of the category of activities to
be regulated under the General permit.
While some of the information
necessary for this evaluation can be
obtained from potential permittees and
others through the proposal of General
permits  for public review, the evaluation
must be completed before any General
permit is issued, and the results must be
published  with the final permit.
  (1) This  evaluation shall be based
upon consideration of the prohibitions
listed in $  230.10(b) and the factors
listed in {  230.10(c), and shall include
documented information supporting
each factual determination in § 230.11 of
the Guidelines (consideration of
alternatives in $ 230.10(a) are not
directly applicable to General permits):
  (2) The evaluation shall include a
precise description of the activities ',o be
permitted  under the General permit.
explaining why they are sufficiently
similar in  nature and in environmental
impact to  warrant regulation under a
single General permit based on Subparis
C-F of the Guidelines. Allowable
differences between activities which
will be regulated under the same
General permit shall be specified.
Activities  otherwise similar in na'ure
may differ in environmental impact due
to their  location in or near ecologically
sensitive areas, areas with unique
chemical or physical characteristics.
areas containing concentrations of toxic
substances, or areas regulated for
specific human uses or by specific land
or water.management plans (e.g..  areas
regulated  under an approved Coastal
Zone Management Plan). If there  are
specific geographic areas within the
purview of a proposed General permit
(called a draft General permit under a
State 404  program), which are more
appropriately regulated by individual
permit due to the considerations cited u>
this paragraph, they shall be clearly
delineated in the evaluation and
excluded  from the permit. In addition
the permitting authority may require an
individual permit for any proposed
activity under a General permit v% here
the nature or location of the activity-
makes an individual permit more
appropriate.
   (3) To predict cumulative effects, 'he
evaluation shall include the number of
individual discharge activities hkeiv '•>
be regulated under a General permit
until its expiration, including repetitions
of individual discharge activities at d
single location.

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  85348
Federal  Register / Vol. 45. No.  249 /  Wednesday. December 24. 1980  / Rules  and  Regulations
  Subpart B—Compliance With th«
  Guideline*

  {230.10 Restriction* on discharge.   _
    Note.—Became other laws may apply to
  particular discharge* and because the Corps
  of Engineers or State 404 agency may have
  additional procedural fad substantive
  requirements, a discharge complying with the
  requirement of these Guidelines will not
  automatically receive a permit
    Although ail requirements in § 230.10
  must be met. the compliance evaluation
  procedures will vary to reflect the
  seriousness of the potential for adverse
  impact* on the aquatic ecosystems
  posed by specific dredged or fill
  material discharge activities.
    (a) Except as provided under
  § 4O4(b)(2). no discharge of dredged or
  fill material shall be permitted if there is
  a practicable alternative to the proposed
  discharge which would have less
  adverse impact on the aquatic
  ecosystem, so long as the alternative
  does not have other-significant adverse
  environmental consequences.
    (1) For the purpose of this
  requirement, practicable alternatives
  include, but are not limited to:
    (i) Activities which do not involve a
  discharge of dredged or Till material into
I  the waters  of the United States or ocean
  waters;
   (ii) Discharges of dredged or fill
  material at other locations in waters of
  the United  States or ocean waters:
   (2) An alternative is practicable if it is
 available and capable of being done
  after taking into consideration cost.
 existing technology, and logistics in light
 of overall project purposes. If it is
 otherwise a practicable alternative, an
 area not presently owned by the
 applicant which could reasonably be
 obtained, utilized, expanded or managed
 in order to fulfill the basic purpose of
 the proposed activity may be
 considered.
   (3) Where the activity associated with
 a discharge which is proposed for a
 special aquatic site (as defined in
 Subpart E) does not require access or
 proximity to or siting within the special
 aquatic site in question to fulfill its basic
 purpose (i.e., is not "water dependent"),
 practicable alternative* that do not
 involve special aquatic sites are
 presumed to be available, unless clearly
 demonstrated otherwise. In addition.
 where a discharge is proposed for a
 special aquatic site, all practicable
 alternative! to the proposed discharge
 which do not involve a discharge into a
 special aquatic cite are presumed to
  iave  lest advene impact on the aquatic
  cosystem, unleta clearly demostrated
'otherwise.
                                  (4) For actions subject to NEPA.
                                where the Corps of Engineers is the
                                permitting agency, the analysis of
                                alternatives required for NEPA
                                environmental documents, including
                                supplemental Corps NEPA documents,
                                will in most cases provide the
                                information for the evaluation of
                                alternatives under these Guidelines. On
                                occasion, these NEPA documents may
                                address a broader range of alternatives
                                than required to be considered under
                                this paragraph or may not have
                                considered the alternatives in sufficient
                                detail to respond to the requirements of
                                these Guidelines. In the latter case, it
                                may be necessary to supplement these
                                NEPA documents with this additional
                                information.
                                  (5) To the extent that practicable
                                alternatives have been identified and
                                evaluated under a Coastal Zone
                                Management program, a { 208 program,
                                or other planning process, such
                                evaluation shall be considered by the
                                permitting authority as part of the
                                consideration of alternatives under the
                                Guidelines. Where such evaluation is
                                less complete than that contemplated
                                under this subsection, it must be
                                supplemented accordingly.
                                  (b) No discharge of dredged or fill
                                material shall be permitted if it:
                                  (1) Causes or contributes, after
                                consideration of disposal site dilution
                                and dispersion, to violations  of any
                               applicable State water quality standard;
                                 (2) Violates any applicable toxic
                               effluent standard or prohibition under
                               section 307 of the Act;
                                 (3) Jeopardizes the continued
                               existence of species listed as
                               endangered or threatened under the
                               Endangered Species Act of 1973, as
                               amended, or results m likelihood of the
                               destruction or adverse modification of a
                               habitat which is determined by the
                               Secretary of Interior or Commerce, as
                               appropriate, to be a critical habitat
                               under the Endangered Species Act of
                               1973, as amended. If an exemption has
                               been granted by the Endangered Species
                               Committee, the terms of such exemption
                               shall apply in lieu of this subparagraph:
                                 (4) Violates any requirement imposed
                               by the Secretary of Commerce to protect
                               any marine sanctuary designated under
                               Title III  of the Marine Protection.
                               Research, and Sanctuaries Act of 1972.
                                 (c) Except as provided under
                               S 404(b)(2), no discharge of dredged or
                               fill material shall be permitted which
                               will cause or contribute to significant
                               degradation of the waters of the United
                               States. Findings of significant
                               degradation related to the proposed
                               discharge shall be based upon
                               appropriate factual determination*.
                               e\ aluations, and tests required by
 Subparts B and G. after consideration of
 Subparts C-F. with special emphasis on
 the persistence and permanence of the
 effects outlined in those Subparts. Under
 these Guidelines, effects contributing to
 significant degradation considered
 individually or collectively, include:
   (1) Significantly adverse effects of the
 discharge of pollutants on human health
 or welfare, including but not limited to
 effects on municipal water supplies,
 plankton, fish, shellfish, wildlife, and
 special aquatic sites.
   (2) Significantly adverse effects of the
 discharge of pollutants on life stages of
 aquatic life and other wildlife dependent
 on aquatic ecosystems,  including the
 transfer, concentration, and spread of
 pollutants or their byproducts outside of
 the disposal site through biological,
 physical, and chemical processes:
   (3) Significantly adverse effects of the
 discharge of pollutants on aquatic
 ecosystem diversity, productivity, and
 stability. Such effects may include, but
 are not limited to,  loss of fish and
 wildlife habitat or loss of the capacity of
 a wetland to assimilate nutrients, punfy
 water, or reduce wave energy; or
   (4) Significantly adverse effects of
 discharge of pollutants on recreational.
 aesthetic, and economic values.
   (d) Except as provided under
 § 404(b)(2). no discharge of dredged or
 fill material shall be permitted unless
 appropriate and practicable steps have
 been taken which  will minimize
 potential adverse impacts of the
 discharge on the aquatic ecosystem.
 Subpart  H identifies such possible steps.

 § 230.11  Factual determination*.
  The permitting authority shall
 determine in writing the potential short-
 term or long-term effects of a proposed
 discharge of dredged or fill material on
 the physical, chemical, and biological
 components of the aquatic environment
 in light of Subparts C-F. Such factual
 determinations shall be used  in S 230.12
 in making findings of compliance or non-
 compliance with the restrictions on
 discharge in §  230.10. The evaluation
 and testing procedures described in
 S 230.60 and § 230.61 of Subpart G shall
 be used as necessary to make, and shall
 be described in. such determination. The
 determinations of effects of each
 proposed discharge shall include the
 following:
  (a) Physical substrate determinations.
 Determine the nature and degree of
effect that the proposed discharge will
have, individually  and cumulatively, on
 the characteristic* of the substrate at
 the proposed disposal site.
Consideration shall be given to the
similarity in particle size, shape, and
degree of compaction of the material

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         Federal  Register / Vol. 45. No.  249 / Wednesday. December 24. 1980 /  Rules and Regulations  85349
 proposed for discharge and the material
 constituting the substrate at the disposal
 site, and any potential changes in
 substrate elevation and bottom
 contours, including changes outside of
 the disposal site which may occur as a
 result of erosion, slumpage, or other
 movement of the discharged material.
 The duration and physical extent of
 substrate changes shall also be
 considered. The possible loss of
 environmental values (§ 230.20] and
 actions to minimize impact (Subpart H)
 shall also be considered in making these
 determinations. Potential changes in
 substrate elevation and bottom contours
 shall be predicted on the basis of the
 proposed method, volume; location, and
 rate of discharge, as well as on the
 individual and combined effects of
 current patterns,  water circulation, wind
 and wave action, and other physical
 factors that may affect the movement of
 the discharged material.
  (b) Water circulation, fluctuation, and
 salinity determinations. Determine the
 nature and degree of effect that the
 proposed discharge will have
 individually and cumulatively on water,
 current patterns, circulation including
 downstream flows, and normal water
 fluctuation. Consideration shall be given
 to water chemistry, salinity, clarity.
 color, odor, taste, dissolved gas levels,
 temperature, nutrients, and
 outrophication ptus other appropriate
 characteristics. Consideration shall also
 be given to the potential diversion or
 obstruction of flow, alterations of
 bottom contours,  or other significant
 changes in the hydrologic regime.
 Additional consideration of the possible
 loss of environmental values (§ 230.23-
 .25) and actions to minimize impacts
 (Subpart H). shall be used in making
 these determinations. Potential
 significant effects on the current
 patterns, water circulation, normal
 water fluctuation and salinity shall be
evaluated on the basis  of the proposed
 method, volume, location, and rate of
discharge.
  (c) Suspended particulate/turbidity
 determinations. Determine the nature
 and degrpe of effect that the proposed
 discharge  will have, individually and
 cumulatively, in terms of potential
changes in the kinds and concentrations
 of suspended particulate/turbidity in the
 vicinity of the disposal site.
 Consideration shall be given to the grain
 size of the material proposed for
 discharge, the shape and size of the
 plume of suspended particulates. the
 duration of the discharge and resulting
 plume and whether or not the. potential
 changes will cause violations of
 applicable water  quality standards.
Consideration should also be given to
the possible loss of environmental
values (S 230.21) -and to actions for
minimizing impacts (Subpart H).
Consideration shall include the
proposed method, volume, location, and
rate of discharge, as well as the
individual and combined effects of
current patterns, water circulation and
fluctuations, wind and wave action, and
other physical factors on the movement
cf suspended particulates.
  (d) Contaminant determinations.
Determine the degree to  which the
material proposed for discharge will
introduce, relocate, or increase
contaminants. This determination shall
consider the material to  be discharged.
the aquatic environment at the proposed
disposal site, and the availability of
contaminants.
  (e) Aquatic ecosystem and organism
determinations. Determine the nature
and degree of effect that the proposed
discharge will have, both individually
and cumulatively, on the structure and
function of the aquatic ecosystem and
organisms. Consideration shall be given
to the effect at the proposed disposal
site of potential changes in substrate
characteristics and elevation,  water or
substrate chemistry, nutrients, currents,
circulation, fluctuation, and salinity, on
the recolonization and existence of
indigenous aquatic organisms  or
communities. Possible loss of
environmental values (5  230.31), and
actions to minimize impacts (Subpart H)
shall be examined. Tests as described in
§ 230.61 (Evaluation and Testing), may
be required to provide information on
the effect of the discharge material on
communities or populations of
organisms expected to be exposed to it.
  (f) Proposed disposal site
determinations. (I) Each disposal site
shall be specified through the
application of these Guidelines. The
mixing zone shall be confined to the
smallest practicable zone within each
specified disposal site that is consistent
with the type of dispersion determined
to be appropriate by the  application of
these Guidelines. In a few special cases
under unique environmental conditions.
where there is adequate  justification to
show that widespread dispersion by
natural means will result in no
significantly adverse environmental
effects, the discharged material may be
intended to be spread naturally in a very
thin layer over a large area of the
substrate rather than be contained
within  the disposal site.
  (2) The permitting authority and the
Regional Administrator  shall consider
the following factors  in determining the
acceptability of a proposed mixing zone:
  (i) Depth of water at the disposal site;
  (ii) Current velocity, direction, and
variability at the disposal sitr.
  (iii) Degree of turbulence;
  (iv) Stratification attributable to
causes such as obstructions, salinity or
density profiles at the disposal site;
  (v) Discharge vessel speed and
direction, if appropriate;
  (vi) Rate  of discharge;
  (vii) Ambient concentration of
constituents of interest;
  (viii) Dredged material characteristics.
particularly concentrations of
constituents, amount of material,  type of
material (sand, silt clay, etc.) and
settling velocities;
  (ix) Number of discharge actions per
unit of time:
  (x) Other factors of the disposal site
that affect the rates and patterns  of
mixing.
  (g) Determination of cumulative
effects on the aquatic ecosystem. (I)
Cumulative impacts are the changes in
an aquatic ecosystem that are
attributable to the collective effect of a
number of individual discharges of
dredged or fill material. Although the
impact of a particular discharge may
constitute a minor change in itself, the
cumulative effect of numerous  such
piecemeal changes can result in a major
impairment of the water resources and
interfere with the productivity  and
water quality of existing aquatic
ecosystems.
  (2) Cumulative effects attributable to
the discharge of dredged or fill material
in waters of the United States should be
predicted to the extent reasonable and
practical. The permitting authority shall
collect information and solicit
information from other sources about
the cumulative impacts on the  aquatic
ecosystem. This information shall be
documented and considered during the
decision-making process concerning the
evaluation  of individual permit
applications, the issuance of a  Genejal
permit, and monitoring and enforcement
of existing permits.
  (h) Determination of secondary
effects on the aquatic ecosystem. (1)
Secondary effects are effects on an
aquatic ecosystem that are associated
with a discharge of dredged or fill
materials, but do not result from  the
actual placement of the dredged or fill
material. Information about secondary
effects on aquatic ecosystems  shall be
considered prior to the time final section
404 action is taken by permitting
authorities.
  (2) Some examples of secondary
effects on an aquatic ecosystem are
fluctuating water levels in an
impoundment and downstream
associated with the operation  of a dam,
septic tank leaching and surface runoff

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 85350  Federal  Register / Vol. 45..No.  249 / Wednesday. December 24, 1980 / Rules  and  Regulations
 from residential or commercial
 developments on Till, and leachate and
 runoff from a sanitary landfill located in
 waters of the US. Activities to be
 r j.iducted on fast land created by the
 ."..schnrjie nf dredged or fill material in
 -•voters of :he United States rr.ay have
 -.econdary impacts with.n !nose 'Alters
 whi-.h should be considered in
 evaluating the impact of creating those
 fast lands.

 § 230.12   Findings of compliance or non-
 contpltanca with the restrictions on
 discharge.
   (ai On the basis of these Guidelines
 (Subparis C through C) the proposed
 disposal Files for the dischaige of
 dredged or fill material must be:
   (1) Specified as complying with the
 requirements of these Guidelines: or
   (2) Specified as complying with the
 requirements of these Guidelines with
 the inclusion of appropriate and
 practicable discharge conditions (se°
 Subpart H) to minimize pollution or  •
 adverse effects to the affected aquatic
 ecosystems; or
   (3) Specified as failing to comply with
 the requirements of these Guidelines
 where:
   (i) There is a pucticdblo alternative to
 the proposed discharge that would have
 less adverse effect on the aquatic
  ccsybtem. so long as such alternative
'does not have other signific.tnt adverse
 environmental consequences; or
   fii) The proposed discharge will result
 in significant degradation of the aquatic
 ecosystem under § 230.10(b) or (c); or
  (Hi) The proposed discharge does not
 include all appropriate and practicable
 measures to minimize potential harm to
 the aquatic ecosystem; or
   (iv) There does not exist sufficient
 information to make a reasonable
 judgment as to whether the proposed
 discharge will comply with these
 Guidelines.
   (b) Findings under  this section shall
 be set forth in writing by the permitting
 authunty for each proposed discharge
 and made available to the permit
 applicant. These f.f dings shall include
 the factual determ -.ations required by
 § 230.11. and a brief explanation of any
 adaptation of these Guidelines to the
 activity under consideration. In the case
of a General permit, such findings shall
be prepared at the time of issuance of
 that permit rather than for each
subsequent discharge under the
authority of that permit.

Subpart C—Potential Impacts on
 Physical and Chemical Characteristics
of the Aquatic Ecosystem

  Note.—The effects described in this
 suboart should be considered in making the
factual determinations and the findings of
compliance or non-compliance in Subpart B.

§230.20  Substrate.
  (aj The substrate of the aquatic
ecosystem underlies open waters of the
United States and constitutes the
surface of wetlands. It consists of
organic and inorganic solid materials
and includes water and other liquids or
gases that fill the spaces between solid
particles.
  (b) Possible loss of environmental
characteristics and values: The
discharge of dredged or fill material can
result in varying degrees of change in
the complex physical, chemical, and
biological characteristics of the
substrate. Discharges which alter
substrate elevation or contours can
result in changes in water circulation.
depth, current pattern, water fluctuation
and water temperature. Discharges mny
adversely affect bottom-dwelling
organisms at the site by smothering
immobile forms or forcing mobile forms
to migrate. Benthic forms present prior
to a discharge are unlikely to recolonize
on the discharged material if it is very
dissimilar from  that of the discharge
sue. Erosion, slumping, or lateral
displacement of surrounding bottom of
such deposits can adversely atfect areas
of the substrate outside the perimeters
of the disposal site by changing or
destroying habitat. The bulk and
composition of the discharged material
and the location, method, and timing of
discharges may all influence the degree
of impact on the substrate.

§ 230.21   Suspended partlculatee/turbldlty.
  (a) Suspended particulates in the
aquatic ecosystem consist of fine-
grained mineral particles, usually
smaller than silt, and organic particles.
Suspended particulates may enter water
bodies as a result of land runoff.
flooding, vegetative and planktonic
breakdown, resuspension of bottom
sediments, and  man's activities
including dredging and filling.
Particulates may remain suspended in
the water column for variable periods of
time as  a result of such factors as
agitation of the  water mass, particulate
specific gravity, particle shape, and
physical and chemical properties of
particle surfaces.
  (b) Possible loss of environmental
characteristics and values: The
discharge of dredged or fill  material can
result in greatly elevated levels of
suspended particulates in the water
column for varying lengths of time.
These new levels may reduce light
penetration and lower the rate of
photosynthesis  and the primary
productivity of an aquatic area if they
last long enough. Sight-dependent
species may suffer reduced feeding
ability leading to limited growth and
lowered resistance to disease if high
levels of suspended particulates persist.
The biological and the chemical content
of the suspended material may react
with the dissolved oxygen in the water.
which can result in e-.jgen depletion.
Toxic metals and organics. pathogens.
and viruses absorbed or adsorbed to
fine-grained particulates in the material
may become biologically available to
organisms either in the water column or
on the substrate. Significant increases in
suspended particulate levels create
turbid plumes which are highly visible
and aesthetically displeasing. The
extent and persistence of these adverse
impacts caused by discharges dap end
upon the relative increase in suspended
particulates above the amount occurring
naturally,  the duration of the higher
levels,  the current patterns, water level.
and fluctuations present when such
discharges occur, the volume, rate, and
duration of the discharge, particulate
deposition, and the seasonal tim.ng of
the disrh  "ge.

§230.22 Water.
  (a) Water is the part of the aqiuhr
ecosystem in which organic and
inorganic constituents are dissolved and
suspended. It constitutes part of the
liquid phase and is contained by the
substrate. Water forms part of a
dynamic aquatic life-supporting system
Water clarity,  nutrients and chemical
content, physical and biological content
dissolved  gas levels, pH, and
temperature contribute to its life-
sustaining capabilities.
  (b) Possible loss of environmental
characteristics and values: The
discharge of dredged or fill material can
change the chemistry and the physical
characteristics of the receiving water at
a disposal site through the introduction
of chemical constituents in suspended or
dissolved form. Changes in the clarity.
color, odor, and taste of water and the
addition of contaminants can reduce or
eliminate  the suitability of water bodies
for populations of aquatic organisms.
and for human consumption, recreation
and aesthetics. The introduction of
nutrients or organic material to the
water column as a result of the
discharge can lead to a high biochen\;td'.
oxygen demand (BOD), which in tui n
can lead to reduced dissolved oxygen
thereby potentially affecting the sur\ •. i.
of many aquatic organisms. Increases -.
nutrients can favor one group of
organisms such as algae  to the dot: ~- •
of other more desirable types such JM
submerged aquatic vegetation.
potentially causing adverse health

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         Federal  Register / Vol. 45. No.  249 / Wednesday.  December 24. 1980 /  Rules and Regulations  85351
 effects, objectionable tastes and odors,
 and other problems.

 § 230.23  Current pattern* and water
 circulation.
   (a) Current patterns and water
 circulation are the physical movements
 of water in the aquatic ecosystem.
 Currents and circulation respond to
 natural forces as modified by basin
 shjpe and cover, phj sira! and chemical
 characteristics of water strata and
 masses, and  energy dissipating factors.
   (b) Possible loss of environmental
 characteristics and values: The
 discharge of  dredged or fill material can
 modify current patterns and water
 circulation by obstructing flow, changing
 the direction or velocity of water flow,
 changing the direction or velocity of
 water flow and circulation, or otherwise
 changing the dimensions of a water
 body. As a result, adverse changes can
 occur in: location, structure,  and
 dynamics of aquatic communities;
 shoreline and substrate erosion and
 depositon rates: the deposition of
 suspended particulates: the rate and
 extent of mixing of dissolved and
 suspended 'Components of the water
 body: and water stratification.

 § 230.24  Normal water fluctuations.
   (a) Normal water fluctuations in a
 natural aquatic system consist of daily,
 seasonal, and annual tidal and flood
 fluctuations in water level. Biological
 and physical  components of such a
 system are either attuned to  or
 characterized by these periodic water
 fluctuations.
  (b) Possible loss of environmental
 characteristics  and values: The
discharge of dredged or fill material can
 alter the normal water-level  fluctuation
 pattern of an  area, resulting in
 prolonged periods of inundation.
 exaggerated extremes of high and low
 water, or a static, nonfluctuating  water
level. Such water level modifications
 may charge salinity patterns, alter
erosion or sedimentation rates.
aggravate water temperature extremes.
and upset the nutrient and dissolved
oxygen balance of the aquatic
ecosystem. In addition, these
 modifications can alter or destroy
communities  and populations of aquatic
animals and vegetation, induce
populations of nuisance organisms,
 modify habitat, reduce food supplies.
 restrict movement of aquatic fauna,
destroy spawning areas, and change
adjacent, upstream, and downstream
 areas.
§230.25  Salinity gradient*.
  (a) Salinity gradients form where salt
water from the-ocean meets and mixes
with fresh water from land.
  (b) Possible loss of environmental
characteristics and values: Obstructions
which divert or restrict flow of either
fresh or salt water may change existing
salinity gradients. For example, partial
blocking of the entrance to an estuary or
river mouth that significantly restricts
the movement of the salt water into and
out of that area can effectively lower the
volume of salt water available for
mixing within that estuary. The
downstream migration of the salinity
gradient can occur, displacing the
maximum sedimentation zone and
requiring salinity-dependent aquatic
biota to adjust to the new conditions.
move to new locations if possible, or
perish. In the  freshwater zone, discharge
operations in the upstream regions can
have equally adverse impacts. A
significant reduction in the volume of
fresh water moving into an estuary
below that which  is considered normal
can affect the location and type of
mixing thereby changing the
characteristic salinity patterns. The
resulting changed circulation pattern
can cause the upstream migration of the
salinity gradient displacing the maximim
sedimentation zone. This migration may
affect those organisms that are adapted
to freshwater environments. It may also
affect municipal water supplies.
  Note.—Possible actions to minimize
adverse impacts regarding site characteristics
c
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85352  Federal  Register / VoL 45: No.  249 / Wednesday. December 24. 1980 /  Rules and Regulations
 •;

 I
  Iensitive to the discharge of material
  uring periods of reproduction and
 growth and development due primarily
 to their limited mobility. They can be
 rendered unfit for human consumption
 by tainting, by production and
 .accumulation of toxir:<: or rjy ingestion
 ,md retention of pathogenic organisms.
 viruses, heavy metals or persistent
 synthetic organic chemicals. The
 discharge of dredged or fill material can
 redirect, delay, or stop the reproductive
 and feeding movements of some species
 of fish and Crustacea, thus preventing
 their aggregation in accustomed places
 such as spawning or nursery grounds
 and potentially leading to reduced
 populations. Reduction of detrital
 feeding species or other representatives
 of lower trophic levels can impair the
 flow of energy from primary consumers
 to higher trophic levels. The reduction or
 potential elimination of food chain
 organism populations decreases the
 overall productmty and nutrient export
 '•jpabihty of the ecosvstem.

 $ 230.32  Other wildlife.
  (a) Wildlife  associated with  aquatic
 fcosystems are resident and transient
 mammals, birds, reptiles, and
 amphibians..
  (0) Possible  loss of values: The
  scharge of dredged or fill material can
 esult in the loss or change of breeding
 ,ir.d nesting areas, escape cover, travel
 corridors, and preferred food sources for
 rebident and transient wildlife  species
 associated with the aquatic ecosystem.
 Those adverse impacts upon wildlife
 h.ibitat  may result from changes  in
 water levels, water flow and circulation.
 salinity, chemical content, and substrate
 characteristics and elevation. Increased
 water U'rbidity can adversely affect
 w.ldlite species which rely upon sight to
 feed, and disrupt the respiration and
 feeding of certain aquatic wildlife and
 food chain organisms. The availability
 of vontaminants  from the discharge of
 dredged or fill  material may lead to the
 bioaccumulation of such contaminants
 in wildlife. Changes in such physical
 and chemical factors of the environment
 may favor the introduction of
 undesirable plant and animal species at
 the expense of resident species and
communities. In some aquatic
environments lowering plant and animal
species diversity may disrupt the normal
functions of the ecosystem and lead to
reductions in overall biological
productivity.
 Note.—Possible jclions to minimize
 I verse impacts  regarding characteristics  of
 blogical components of the aquatic
 icosystem can be found in Subpart H.
 Subpart E—Potential Impacts on
 Special Aquatic Sites

   Note.—The impacts described in this
 subpart should be considered in making the
 factual determinations and the findings of
 compliance or non-compliance in Subpart B
 The definition of special aquatic, sites is
 found in § 2303(q-l).

 § 230.40 Sanctuaries, and reluyea.
   (a) Sanctuaries and refuges consist cf
 areas designated under State and
 Federal laws or local ordinances to be
 managed principally for the preservation
 and use of fish and wildlife resources.
   (b) Possible loss of values:
 Sanctuaries and refuges may be affected
 by discharges of dredged or fill material
 which will:
   (1)  Disrupt the breeding, spawning.
 migratory movements or other critical
 life requirements  of resident or transient
 fish and wildlife resources:
   (2)  Create unplanned, easy and
 incompatible human access to remote
 aquatic areas:
   (3)  Create the need for frequent
 maintenance activity;
   (4) Result in the establishment of
 undesirable competitive species of
 plants and animals:
   (5) Change the balance of water and
 land areas needed to provide cover.
 food,  and other fish  and wildlife habitat
 requirements in a way that modifies
 sanctuary or refuge  management
 practices:
   (6) Result in any of the other adverse
 impacts discussed in Subparts C and D
 as they relate to a particular sanctuary
 or refuge.

 § 230.41 Wetlands.
   (a)[l) Wetlands consist of areas that
 are inundated or saturated by surface or
 ground water at a frequency and
 duration sufficient to support, and that
 under normal circumstances do support.
 a prevalence of vegetation typically
 adapted for life in saturated soil
 conditions.
  (2) Where wetlands are adjacent to
 open water, they generally constitute the
 transition to upland. The margin
 between wetland  and open water can
 best be established by specialists
 familiar with the local environment,
 particularly where emergent vegetation
 merges with submerged vegetation  over
 a broad area in such places as the
 lateral margins of open water,
 headwaters, rainwater catch basins, and
groundwater seeps. The landward
margin of wetlands also can best be
 identified by specialists familiar with
 the local environment when vegetation
 from the two regions merges over a
 broad area.
  (3) Wetland vegetation consists of
plants that require saturated soils to
survive (obligate wetland plants) as well
as plants, including certain trees, that
gam a  competitive advantage over
others because they can tolerate
prolonged wet soil conditions and their
competitors cannot. In addition to plant
populations and communities, wetlands
are delimited by hydrological and
physical characteristics of the
environment. These characteristics
should be considered when information
about them is needed to supplement
information available about vegetation.
or where wetland vegetation has been
removed or is dormant.
  (b) Possible loss of values: The
discharge of dredged or Till material in
wetlands is likely to damage or destroy
habitat and adversely affect  the
biological productivity of wetlands
ecosystems by smothering, by
dewatering. by permanently  flooding, or
by altering substrate elevation or
periodicity of water movement. The
addition of dredged or fill jnaterial may
destrov wetland vegetation or result in
advancement of succession to dry land
species. It may reduce or eliminate
nutrient exchange by a reduction of the
system's productivity,  or by altering
current patterns and velocities.
Disruption or elimination of the wetland
system can degrade water quality by
obstructing circulation patterns that
flush large expanses of wetland
systems, by interfering with the
filtration function of wetlands, or by
changing t) » aquifer recharge capability
of a wetland. Discharges can also
change the wetland habitat value for
fish and wildlife as discussed in Subpart
D. When disruptions in flow  and
circulation patterns occur, apparently
minor loss of wetland acreage may
result in major losses through secondary
impacts. Discharging fill material in
wetlands as part of municipal, industrial
or recreational development  may modify
the capacity of wetlands to retain and
store floodwaters and to serve as a
buffer zone shielding upland  areas from
wave actions, storm damage  and
erosion.
§230.42  Mudflats
  (a) Mud flats are broad flat areas
along the sea coast and in coastal rivers
to the head of tidal influence and in
inland lakes, ponds, and riverine
systems. When mud flats are inundated.
wind and wave action may resuspend
bottom sediments. Coastal mud flats are
exposed at extremely low tides and
inundated at high tides with  the water
table at or near the surface of the
substrate. The substrate of mud flats
contains organic material and particles

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         Federal Register  /  Vol. 45.  No. 249 / Wednesday.  December 24. 1960  /  Rulea and Regulations  85353
 smaller in size than sand. They are
 either unvegetated or vegetated only by
 algal mats.
   (b) Possible loss of values: The
 discharge of dredged or fill material can
 cause changes in water circulation
 patterns which may permanently flood
 or dewater the mud flat or disrupt
 periodic inundation,  resulting in an
 increase in the rate of erosion or
 accretion. Such changes can deplete or
 eliminate mud flat biota, foraging areas.
 ji.d nursery areas. Changes in
 inundation patterns can affect the
 chemical and biological exchange and
 decomposition process occurring on the
 mud flat and charge the deposition of
 suspended material affecting the
 productivity of the area. Changes may
 reduce the mud flat's capacity to
 dissipate storm surge runoff.
 § 230.43  Vegetated (hallow*.
   (a) Vegetated shallows are
 permanently inundated areas that under
 normal circumstances support
 communities of rooted aquatic
 vegetation, such as turtle grass and
 eelgrass  in estuarine or marine systems
 as well as a number of freshwater
 species in rivers and lakes.
  (b) Possible loss of values: The
 discharge of dredged or fill material can
 smother  vegetation and benthic
 organisms, it may also create unsuitable
 conditions for their continued vigor by:
 (1) changing water circulation patterns:
 (2} releasing nutrients that increase
 undesirable algal populations; (3)
releasing chemicals that adversely
 affect plants and animals; (4) increasing
 turbidity levels, thereby reducing light
penetration and hence photosynthesis:
 and (5) changing the capacity of a
vegetated shallow to stabilize bottom
materials and decrease channel
shoaling. The discharge of dredged or
fill material may reduce the value of
vegetated shallows as nesting.
spawning, nursery, cover, and forage
areas, as well as their value in
protecting shorelines from erosion and
wave actions. It may also encourage the
growth of nuisance vegetation.

§230.44  Coral reef*.
  (a) Coral reefs consist of the skeletal
deposit, usually of calcareous or
silicaceous materials, produced by the
vital activities of anthozoan polyps or
other invertebrate organisms present in
growing portions of the reef.
  (b) Possible loss of values: The
discharge of dredged or fill material can
adversely affect colonies of reef building
organisms by burying them, by releasing
contaminants such as hydrocarbons into
 the water column, by reducing light
penetration through the water, and by
increasing the level of suspended
particulates. Coral organisms are
extremely sensitive to even slight
reductions in light penetration or
increases in suspended particulates.
These adverse effects will cause a loss
of productive colonies which in turn
provide habitat for many species of
highly specialized aquatic organisms.

§ 230.45  Riffle and pool complexes.
  (n) Steep gradient sections of streams
are sometimes characterized by riffle
and pool complexes. Such stream
sections are recognizable by their
hydraulic characteristics. The rapid
movement of water over a coarse
substrate in riffles results in a rough
flow, a turbulent surface, and high
dissolved oxygen levels in the water.
Pools are deeper areas associated with
riffles. Pools are characterized by a
slower stream velocity, a steaming flow,
a smooth surface, and a finer substrate.
Rifle and pool complexes are
particularly valuable habitat for fish and
wildlife.
  (b) Possible loss of values: Discharge
of dredged or fill material can eliminate
riffle and pool areas by displacement,
hydrologic modification, or
sedimentation. Activities which affect
riffle and pool areas and especially
riffle/pool ratios,  may reduce the
aeration and filtration capabilities at the
discharge site and downstream, may
reduce stream habitat diversity, and
may retard repopulation'of the disposal
site and downstream waters through
sedimentation and the creation of
unsuitable habitat. The discharge of
dredged or fill material which alters
stream hydrology may cause scouring or
sedimentation of riffles and pools. *
Sedimentation induced through
hydrological modification or as a direct
result of the deposition of
unconsolidated dredged or fill material
may clog riffle and pool areas,  destroy
habitats, and create anaerobic
conditions. Eliminating pools and
meanders by  the discharge of dredged or
fill material can reduce water holding
capacity of streams and cause  rapid
runoff from a watershed. Rapid runoff
can deliver large quantities  of flood
water in a short time to downstream
areas resulting in the destruction of
natural habitat, high property loss, and
the need for further hydraulic
modification.
  Note.—Possible actions to minimize
advene impacts on tile or material
characteristics can be found in Subpart H.
Subpart F—Potential Effects on
Human Use Characteristics

  Note.—The effects described in this
subpart should be considered in making the
factual determinations and the findingi of
compliance or non-compliance in Subpart B.

§ 230.50 Municipal and private water
supplies.
  (a) Municipal and private water
supplies consist of surface water or
ground water which is directed to  '.he
intake of a municipal or private water
supply system.
  (b) Possible loss of values: Discharges
can affect the quality of water supplies
with respect to color, taste, odor.
chemical content and suspended
particulate concentration, in such a way
as to reduce the fitness of the water  for
consumption. Water can be rendered
unpalatable or unhealthy by the
addition of suspended particulates.
viruses and pathogenic organisms, and
dissolved  materials. The expense of
removing such substances before the
water is delivered for consumption can
be high. Discharges  may also affect the
quantity of water available  for
municipal and  private water sapplies In
addition, certain commonly used water
treatment chemicals have the potential
for combining with some suspended or
dissolved substances from dredged or
fill material to  form  other products that
can have a toxic effect  on consumers

§230.51  Recreational and commercial
fisheries.
  (a) Recreational and  commercial
fisheries consist of harvestable fish.
crustaceans, shellfish, and other aquatic
organisms used by man.
  (b) Possible loss of values: The
discharge of dredged or fill materials
can affect the suitability of recreational
and commercial fishing grounds as
habitat for populations of consumable
aquatic organisms. Discharges can result
in the chemical contamination of
recreational or commercial  fisheries.
They may also interfere with the
reproductive success of recreational and
commercially important aquatic species
through disruption of migration and
spawning areas. The introduction of
pollutants at critical times in their life
cycle may directly reduce populations cf
commercially important aquatic
organisms or indirectly reduce them by
reducing organisms upon which they
depend for food. Any of these impacts
can be of short duration or prolonged.
depending upon the physical and
chemical impacts of the discharge and
the biological availability of
contaminants to aquatic organisms.

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       85354  Federal Register  /  Vol. 45. No.  249 / Wednesday. December 24. 1980  /  Rules  and  Regulations
      n
•
   230.52  Water-related recreation.
   (a) Water-related recreation
 encompasses activities undertaken for
 amusement and relaxation. Activities
 eniompass two broad categories of use:
 consumptive, e.g.. harvesting resources
 by hunting and fishing; and non-
 u-Tfjmplive. e g. canoe-ny and sight-
 SCl T.S.
   ;hj Possible loss of values: One of the
 more important direct impacts of
 dredged or fill disposal is to impair or
 destroy the resources which support
 recreation activities. The disposal of
 dredged or fill material may adversely
 rr. odify or destroy water jse for
 recreation by changing turbidity.
 suspended particulates. temperature.
 dissolved oxygen, dissolved materials.
 toxic materials, pathogenic organisms,
 quality of habitat, and the aesthetic
 qualities of sight, taste, odor, and color.

 5 230.53  Aesthetics.
   Id) Aesthetics associated  with the
 ,. .j-.ia'ic ecosystem consist of ths
 perception of beauty by one or a
 roT.hination of the senses of sight.
 r.earng, touch, and smell  Aesthetics of
 juuatic ecosystems apply to '.he quality
 of iife enjoyed by the genera! public and
 property owners.
   P(b) Possible loss of values: The
 ischarge of dredged or fill material can
 :ar the beauty of natural aquatic
 i-ovvstems by degrading water quality,
 i. resting distracting disposal sites,
 •r.d'irmg inappropriate development.
 t eicoiiragirg unplanned and
 incompatible human access, and by
 iJostroying vital elements that contribute
 to the compositional  harmony or unity,
 \ isual distinctiveness. or diversity of an
 ai  ea. The discharge of dredged or fill
 material can adversely affect the
 particular features, traits, or
 characteristics of an  aquatic area which
 Tv-ke it valuable to property owners.
 Activities which degrade water quality,
 disrupt natural substrate and
 vrgptational characteristic*, deny
 jccrss to or visibility of the resource, or
 >vs':it in changes in odor, air quality, or
 noise levels may reduce the value of an
 .-iquatic area to private property owners.

 §230.54  Parks, national and historical
 monument*, national seashores, wHdemess
areas, research sites, and similar
preserves.
  (a)  These preserves consist of areas
designated under Federal and State
 la AS  or local ordinances to be managed
for their aesthetic, educational.
histoiical, recreational, or scientific
value.
  (b)  Possible loss of values: The
 ischarge of dredged or fill material into
such areas may modify the aesthetic.
 educational, historical, recreational*
 and/or scientific qualities thereby
 reducing or eliminating the uses for
 which such sites are set aside and
 managed.
  Note.—Possible actions tc minimize
 adverse .mpacls regarding site or material
 (.haracteristics can be found in Subpart H.

 Subpart G—Evaluation and Testing

 § 230.60  General evaluation of dredged or
 (ill material.
  The purpose of these evaluation
 procedures and the chemical and
 biological testing sequence outlined in
 § 230.61 is to provide information  to
 reach the determinations required by
 § 230.11. Where  the results of prior
 evaluations, chemical and biological
 tests, scientific research, and experience
 can provide information helpful in
 making a determination, these should be
 used. Such prior results may make new
 testing unnecessary. The information
 used shall be documented. Where the
 same information applies to more  than
 one determination, it may be
 documented once and referenced  in
 later determinations.
  (a) If the evaluation under paragraph
 (b) indicates the dredged or fill material
 is not a carrier of contaminants, then the
 required determinations pertaining to
 the presence and effects of
 contaminants can be made without
 testing. Dredged or fill material is  most
 likely to be free from chemical,
 biological, or other pollutants where it is
 composed primarily of sand, gravel, or
 other naturally occurring inert material.
 Dredged material so composed is
generally found in areas of high current
 or wave energy such as streams with
large bed loads or coastal areas with
 shifting bars and channels. However,
 when such material is discolored or
 contains other indications that
contaminants may be present, further
 inquiry should be made.
  (b) The extraction site shall be
 examined in order to assess whether it
 is sufficiently removed from sources of
pollution to provide reasonable
assurance that the proposed discharge
material is not a carrier of
contaminants. Factors to be considered
include but are not limited to:
  [1] Potential routes of contaminants or
contaminated sediments to the
extraction site, based on hydrographic
or other maps, aerial photography, or
other materials that show watercourses,
surface relief, proximity to tidal
movement, private and public roads.
location of buildings, municipal and
industrial areas, and agricultural or
forest lands.
  (2) Pertinent results from tests
previously carried out on the material at
the extraction site, or carried out on
similar material for other permitted
projects in the vicinity. Materials shall
be considered similar if the sources of
contamination,  the physical
configuration of the sites and the
sediment composition of the materials
are comparable, in light of water
circulation and stratification, sediment
accumulation and general sediment
characteristics. Tests from other sites
may be relied on only if no changes
have occurred at the extraction sites  to
render the results irrelevant.
  (3) Any potential for significant
introduction of persistent pesticides
from land runoff or percolation:
  (4) Any records of spills or disposal of
petroleum products or substances
designated as hazardous under section
311 of the Clean Water Act (See 40 CFR
116);
  (5) Information in Federal. State and
local records indicating significant
introduction of pollutants from
industries, municipalities, or other
sources, including types and amounts cf
wast-.- t..^terials discharged along the
potential routes of contiminants to the
extraction site: and
  (6) Any possibility of the presence  of
substantial natural  deposits of minerals
or other substances which could be
released to the aquatic environment in
harmful quantities by man-induced
discharge activities.
  (c) To reach the determinations in
§ 230.11 involving potential effects of the
discharge on the characteristics of the
disposal site, the narrative guidance in
Subparts C-F shall be used along with
the general evaluation procedure in
5 230.60 and. if necessary, the chemical
and biological testing sequence in
§ 230.61. Where the discharge site is
adjacent to the extraction site and
subject to the same sources of
contaminants, and materials at the two
sites are substantially similar, the fact
that the material to be discharged may
be a carrier of contaminants is not likely
to result in degradation of the disposal
site. In such circumstances, when
dissolved material and suspended
particulates can be controlled to prevent
carrying pollutants  to less contaminated
areas, testing will not be required.
  (d) Even if the § 230.60(b) evaluation
(previous tests, the  presence of polluting
industries and information about their
discharge or runoff into waters of the
U.S., bioinventories. etc.) leads to the
conclusion that there is  a high
probability that the material proposed
for  discharge is a carrier of
contaminants, testing may not be
necessary if constraints are available to

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         Federal  Register / Vol. 45. No.  249 /  Wednesday. December 24. 1980 / Rules  and  Regulations   83355
 reduce contamination to acceptable
 levels within the disposal site and to-
 prevent contaminants from being
 transported beyond the boundaries of
 the disposal site, if such constraints are
 acceptable to the permitting authority
 and the Regional Administrator, and if
 the potential discharger is willing and
 able to implement such  constraints.
 However, even if tests are not
 performed, the permitting authority must
 still determine the probable impact of
 the operation on the receiving aquatic
 ecosystem. Any decision not to test
 must be explained in the determinations
 made under § 230.11.

 § 230.61  Chemical, biological, and physical
 •valuation and testing.
  Note.—The Agency is today proposing
 revised testing guidelines. The evaluation and
 testing procedures in this section are based
 on the 1975 i 404(b)(l) interim final
 Guidelines and shall remain in effect until the
 revised testing guidelines are published as
 fir.dl regulations.
  (a) No single test or approach can be
 applied in all cases to evaluate the
 effects of proposed discharges of
 dredged or fill materials. This section
 provides some guidance in. determining
 which test and/or evaluation procedures
 are appropriate in a given case. Interim
 guidance to applicants concerning the
 applicability of specific approaches or
 procedures will be furnished by the
 permitting authority.
  (b) Chemical-biological interactive
 ejects. The principal concerns of
 discharge of dredged or  fill material  that
 contain contaminants are the potential
 effects on the water column and on
 communities of aquatic organisms.
  (I) Evaluation of chemical-biological
interactive effects. Dredged or fill
material may be excluded from  the
evaluation procedures specified in
paragraphs (b)(2) and (3) of this section
if it is determined, on the basis of the
evaluation in § 230.60. that the
l.kelihood of contamination by
contaminants is acceptably low. unless
 the permitting authority, after evaluating
and considering any comments received
 from the Regional Administrator.
determines that these procedures are
 necessary. The Regional Administrator
may require, on a case-by-case basis,
 testing approaches and procedures by
stating what additional information is
needed through further analyses and
how the results of the analyses will be
of value in evaluating potential
environmental effects.
  If the General Evaluation indicates the
presence of a sufficiently large number
of chemicals to render impractical the
 identification of all contaminants by
chemical testing, information may be
obtained from bioassays in lieu of
chemical tests.
  (2)  Water column effects, (i)
Sediments normally contain constituents
that exist in various chemical forms and
in various concentrations in several
locations within the sediment. An
elutriate teat may be used to predict the
effect on water quality due to release of
contaminants from the sediment to the
water column. However, in the case of
fill material  originating on land which
may be a carrier of contaminants, a
water leachate test is appropriate.
  (ii) Major constituents to be analyzed
in the elutriate are those deemed critical
by the permitting authority, after
evaluating and considering any
comments received from the Regional
Administrator, and'considering results
of the evaluation in { 230.60. Elutriate
concentrations should be compared to
concentrations of the same constituents
in water from the disposal site. Results
should be evaluated in light of the
volume and rate of the intended
discharge, the type of discharge, the
hydrodynamic regime at the disposal
sitp. and other information relevant to
the impact on water quality. The
permitting authority should consider the
mixing zone in evaluating water column
effects. The permitting authority may
specify bioassays when such procedures
will be of value.
  (3) Effects on benthos. The permitting
authority may  use an appropriate
benthic bioassay (including
bioaccumulation tests) when such
procedures will be of value in assessing
ecological effects and in establishing
discharge conditions.
  (c) Procedure for comparison of sites.
  (t) When an inventory of the total
concentration of contaminants would be
of value in comparing sediment at the
dredging site with sediment at the
disposal site, the permitting authority
may require a sediment chemical
analysis. Markedly different
concentrations of contaminants between
the excavation and disposal sites may
aid in making an environmental
assessment of the proposed disposal
operation. Such differences should be
interpreted in terms of the potential for
harm  as supported by any pertinent
scientific literature.
  (2) When an analysis of biological
community structure will be of value  to
assess the potential for adverse
environmental impact at the proposed
disposal site, a comparison of the
biological characteristics between  the
excavation and disposal sites may be
required by  the permitting authority.
Biological indicator species may be
useful in evaluating the existing degree
of stress at both sites. Sensitive species
representing community components
colonizing various substrate types
within the sites should be identified as
possible bioassay organisms if tests for
toxicity are required. Community
structure studies should be performed
only when they will be of value in
determining discharge conditions. This
is particularly applicable to large
quantities of dredged material known to
contain adverse quantities  of toxic
materials. Community studies should
include benthic organisms such as
microbiota and harvestable shellfish
and finfish. Abundance, diversity, and
distribution should be documented and
correlated with substrate type and other
appropriate physical and chemical
environmental characteristics.
  (d) Physical tests and evaluation. The
effect of a discharge of dredged or fill
material on physical substrate
characteristics at the disposal site, as
well as on the water circulation.
fluctuation, salinity, and suspended
particulates content there, is important
in making factual determinations  in
§ 230.11. Where information on such
effects is not  otherwise available  to
make these factual determinations, the
permitting authority shall require
appropriate physical tests and
evaluations as are justified and deemed
necessary. Such tests  may  include sieve
tests, settleability tests, compaction
tests, mixing zone and suspended
particulate plume determinations, and
site assessments of water flow.
circulation, and salinity characteristics.

Subpart H—Actions To Minimize
Adverse Effects

  Note.—There are many actions which can
be undertaken  in response to § 203 10|d) lu
minimize the adverse effects of discharges of
dredged or fill material. Some  of these.
grouped by type of activity, are listed in th-s
subpart.

§ 230.70  Actions concerning the location
of the discharge.
  The effects of the discharge can be
minimized by the choice of the disposal
site. Some of the ways to accomplish
this  are by:
  (a) Locating and confining the
discharge to minimize smothering of
organisms:
  (b) Designing the discharge to avoid  a
disruption of periodic water inundation
patterns:
  (c) Selecting  a disposal site that has
been used previously for dredged
material discharge;
  (d) Selecting a disposal site at which
the substrate is composed  of material
similar to that being discharged, such as
discharging sand on sand or mud on
mud:

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 85356  Federal Register  /  Vol. 45. No. 249 / Wednesday.  December 24. 1980 /  Rules and Regulations
    [>•) Selecting (he disposal site, the
 'discharge point, and the method of
 discharge to minimize the extent of any
 pKirne:
    If) Designing the discharge of dredged
 01 fill material lo minimize or prevent
 Jp.e rn ution of standing bodies pf water
 IP ,.ieds of normally fluctuating wjter
 li".ds H,-.d minimize or prevent the
 •ir IIP !2e of dredS ••'ubject to such
 fluctuations.

 $ 230.71   Actions concerning the material
 to be discharged.
   The effects of a discharge can be
 minimiz?d by treatment of. or
 limitations on the material itself, such
   (d) D.sposal of Jredged material in
 si.ch d manner that physiochemical
 conditions are maintained and the
 potency and availability of pollutants
 are reduced.
   (b) Limiting the solid, liquid, and
 gaseous components of material to be
 discharged at a particular site;
   (c) Adding treatment substances to
 'he discharge material:
   (d) Utilizing chemical flocculants to
 ' nmince fhe deposition of suspended
 p.-rticu!.ites in diked  disposal areas.

 ; 230.72 Actions controlling the material
 after discharge.
   The effects of the dredged or fill
  idterial after discharge may be
    lrolled by:
   (a) Selecting discharge methods and
 disposal sites where the potential for
 erosion, slumping or leaching of
 rn.itenals into  the surrounding aquatic
 11 usvsiem will be reduced. These sites
 or methods include, but are not limited
 :.)•
  (1) L'smg containment levees, sediment
 hdsins. and cover crops to  reduce
 erosion;
  (2} Using lined containment areas to
 i educe leaching where leaching of
 ..hemical constituents from the
 discharged material is expected to be a
 problem:
  (b) Capping in-place contaminated
 material with clean material or
 selectively discharging the most
 contaminated material first to be capped
 with the remaining material:
  [c) Maintaining and containing
 discharged material properly to prevent
 point and nonpoint sources of pollution:
  (d) Timing the discharge to minimize
 impact, for instance during periods of
 unusual high  water flows, wind. wave.
 and tidal actions.

 § 230.73  Actions affecting the method  of
Dispersion.
  The effects of a discharge can be
  inimized by the manner in which it  is
'dispersed, such as:
   (a) Where environmentally desirable.
 distributing the dredged material widely
 in a thin layer at the disposal site to
 maintain natural substrate contours and
 elevation;
   (b) Orienting a dredged or fill rr.d'.eiial
 mound to minimize undesirable
 obstruction to the water current or
 cirrnldtion pattern, and utilizing natural
 boiton contours to minimize 'he s:zu of
 the mound:
   (c) Using silt screens or other
 appropriate methods to confine
 suspended participate/turbidity to a
 small area where settling or removal can
 occur
   (d) Making use of currents and
 circulation patterns to mix, disperse and
 dilute the discharge:
   (e) Minimizing water column turbidity
 by using a submerged diffuser system. A
 similar effect can be accomplished by
 submerging pipeline discharges or
 otherwise releasing materials near the
 bottom;
   (0 Selecting sites or managing
 discharges to confine and minimize the
 release of suspended partic ulates to give
 decreased turbidity levels and to
 m.untam light penetration for organisms:
   (g) Setting limitations on the amount
 of material to be discharged per unit of
 time or volume of receiving water.

 § 230.74  Actions related to technology.
  Discharge technology should be
 adapted to the needs of each site. In
 determining whether the discharge
 operation sufficiently minimizes adverse
 environmental impacts, the applicant
 should consider:
  (a) Using appropriate equipment or
 machinery, including protective devices.
 and the use of such equipment or
 machinery in activities related to the
 discharge of dredged or fill material:
  (b) Employing appropriate
 maintenance and operation on
 equipment or machinery, including
 adequate training, staffing, and working
 procedures;
  (c) Using machinery and techniques
 that are especially  designed to reduce
 damage to wetlands. This may include
 machines equipped with devices that
 scatter rather than mound excavated
 materials, machines with specially
designed wheels or tracks, and the use
of mats under heavy machines to reduce
 wetland surface compaction and rutting;
  (d) Designing access roads and
channel spanning structures using
culverts, open channels, and diversions
 that will pass both low and high water
flows, accommodate fluctuating water
levels, and maintain circulation and
 faunal movement;
  (e) Employing appropriate machinery
and methods of transport of the material
for discharge.

§ 230.75  Actions affecting plant and
animal populations.
  Minimization of adverse effects on
populations of plants and animals can
!jo dchieved by:
  (.i) Avoiding chdPges in water current
dnd Circulation patterns which '.vould
interfere with the movement of animals:
  (b) Selecting sites or managing
discharges to prevent or avoid creating
habitat conducive to the development of
undesirable predators or species which
have a competitive edge ecologically
over indigenous plants or animals;
  (c) Avoiding sites having unique
habitat or other value, including habitat
of threatened or endangered species:
  (d) Using planning and construction
practices to institute habitat
development and restoration to produce
a new or modified environmental state
of higher ecological value by
displacement of some or all of the
existing environmental characteristics.
Habitat development and restoration
techniques can be used to minimize
adverse impacts and to compensate for
destroyed habitat. Use techniques that
have been demonstrated to be effective
in circumstances similar to  those under
consideration wherever possible. Where
proposed development and restoration
techniques have not yet advanced to the
pilot demonstration stage, initiate  their
use on a small scale 'o allow corrective
action if unanticipated adverse impacts
occur.
  (e) Timing discharge to avoid
spawning or migration seasons and
other biologically critical time  periods:
  {f) Avoiding the destruction of
remnant natural sites within areas
already affected by development.

§ 230.76  Action* affecting human use.
  Minimization of adverse effects on
human use potential may be achieved
by:
  (a) Selecting discharge sites  and
following discharge procedures to
prevent or minimize any potential
damage to  the aesthetically pleasing
features of the aquatic site (e.g.
v-iewscapes), particularly with respect to
water quality;
  (b) Selecting disposal sites which are
not valuable as natural aquatic areas;
  (c) Timing the discharge to avoid the
seasons or periods when human
recreational activity associated with the
aquatic site is most important:
  (d) Following discharge procedures
which avoid or minimize the disturbance
of aesthetic features of an aquatic site or
ecosystem.

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         Federal  Register /  Vol.  45.  No. 249  /  Wednesday.  December 24, 1980  /  Rules  and  Regulations  85357
   (e) Selecting sites that will not be
 detiimental or increase incompatible
 human activity or require the need for
 frequent dredge or fill maintenance
 activity m remote fish and wildlife
 areas.
   (f) Locating the disposal site outside
 of !hp vic:n t\ of a public water -.apply
 intake

 § 230 77  Other actions.
   (a; In the case of fills, controlling
 runoff and other discharges from
 activities to be conducted on the fill:
   (b) In the case of dams, designing
 watrr releases to accommodate the
 needs of Fish and wildlife.
   (c) In dredging projects funded by
 Federal agencies other than the Corps of
 Engineers, maintain desired water
 quality of the return discharge through
 agreement with the Federal  funding
 authority on scientifically defensible
 pollutant concentration levels in
 addition to any applicable water quality
 standards.
   (d) When a significant ecological
 change in the aquatic environment is
 proposed by the discharge of dredged or
 fill material, the permitting authority
 should consider the ecosystem that will
 be lost as well as the environmental
 benefits of the new system.

 Subpart I—Planning To Shorten Permit
 Processing Time

 § 230.80  Advanced identification of
 disposal areas.
  (a) Consistent with  these Guidelines,
 F.PA and the permitting authority, on
 their own initiative or at the request of
 any other party and after consultation
 with any-affected State that is not the
permitting authority, may identify sites
 which  will be considered as:
  (1) Possible future disposal sites,
 including existing disposal sites and
non-sensitive areas; or
  (2) Areas generally  unsuitable for
disposal site specification:
  (b) The identification of any area as a
 possible future disposal site should not
be deemed to constitute a permit for the
discharge of dredged of fill material
 within such area or a specification of a
disposal site. The identification of areas
 that generally will not be available for
 disposal site specification should not be
 deemed as prohibiting applications for
 permits to discharge dredged or fill
 material in such areas. Either type of
 identification constitutes information  to
 facilitate individual or General permit
 application and processing.
   (c) An appropriate public notice of the
 proposed identification of such arias
 shall be issued:
   (d) To provide the basis for advanced
 identification of disposal areas, and
 areas unsuitable for disposal, EPA and
 the permitting authority shall consider
 the likelihood that use of the area in
 question for dredged or fill material
 disposal will comply with these
 Guidelines. To facilitate this analysis.
 EPA and the permitting authority should
• review available water resources
 management data including data
 available from the public, other Federal
 and State agencies, and information
 from  approved Coastal Zone
 Management programs and River Basin
 Plans.
   (e)  The permitting authority should
.maintain a public record of the
 identified areas and a  written statement
 of the basis for identification.
 |FR Doc 80-W001 Filld 12-23-8O 545 ami
 BILLING COOC (MO-01-V

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       APPENDIX B
GUIDANCE FOR EVALUA TJON
 OF EFFLUENT DISCHARGES
 FROM CONFINED DISPOSAL
       FACILITIES
                              DRAFT

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DRAFT

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                           TABLE OF CONTENTS
                                                                          Page No.

Table of Contents                                                                   i
List of Tables                                                                     Hi
List of Figures                                                                    iv
APPENDIX B -      GUIDANCE FOR EVALUATION OF EFFLUENT DISCHARGES
                   FROM CONFINED DISPOSAL FACILITIES  	  B-l

Bl.O        INTRODUCTION	  B-l
Bl.l        Background	  B-l
B1.2        Purpose and Scope	  B-3
B1.3        Regulatory Considerations  	  B-3
B1.4        Applicability	  B-4
Bl.4.1       Hydraulic Filling	  B-4
Bl.4.2       Flow Through Dikes   	  B-5
Bl.4.3       Mechanical Filling	  B-5
Bl.4.4       Surface Runoff and Leachate	  B-5

B2.0        OVERVIEW OF EVALUATIONS FOR EFFLUENT DISCHARGES	  B-6
B2.1        Water Quality Standards  	  B-6
B2.2        Mixing Zones	  B-6
B2.3        Basis of Evaluations	  B-7
B2.4        Contaminant Controls  	  B-7
B2.5        Tiered Approach	  B-7
B2.6        Tier II: Water Quality Evaluations  	  B-8
B2.6.1       Screen Relative to WQS	  B-8
B2.6.2       Testing for Evaluation of Effluent Water Quality  	B-10
B2.7        Tier III:  Toxicity Evaluations	B-ll

B3.0        TESTING PROCEDURES FOR EFFLUENT DISCHARGES 	B-ll
B3.1        Data Requirements  	B-12
B3.1.1       Disposal Area Design	B-12
B3.1.2       Sampling Requirements  	B-14
B3.2        Column Settling Tests  	B-14
B3.2.1       Apparatus	B-15
B3.2.2       Test Procedure	B-15
B3.2.3       Data Analysis	B-18
B3.2.4       Determination of Effluent Suspended Solids Concentration  	B-19
B3.2.5       Determination of Field Mean Retention Time	B-21
B3.3        Modified Elutriate Test Procedure  	B-22
B3.3.1       Apparatus	B-22
                                                                     DRAFT

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                                           11

B3.3.2       Test Procedure	B-23
B3 3 .3       Chemical Analyses	B-25
B3.3.4       Dissolved Concentrations of Contaminants	B-25
B3.3.5       Calculation of Total Concentrations of Contaminants	B-25
B3.4         Water Column Toxicity Test Procedure	B-26
B3.4.1       Apparatus	B-26
B3.4.2       Test Procedure	B-27

B4.0         EXAMPLE CALCULATIONS	B-29
B4.1         Example  1:   Evaluation of  Effluent  Water  Quality For  an Existing
             Disposal Area	B-29
B4.1.1       Project Information	B-29
B4.1.2       Modified Elutriate Testing  	B-29
B4.1J       Column Settling Tests  	B-30
B4.1.4       Prediction  of Effluent Suspended Solids Concentration	B-33
B4.1.5       Prediction  of Contaminant Concentrations	B-33
B4.2         Example 2:   Determination  of Disposal Area  Requirements to Meet  a
             Given Effluent Quality Standard 	B-34
B4.2.1       Project Information	B-34
B4.2.2       Modified Elutriate Testing  	B-34
B4.2.3       Column Settling Tests  	B-34
B4.2.4       Determination of Allowed Effluent Suspended Solids Concentrations	B-35
B4.2.5       Determination of Ponded Volume and Surface Area	B-36

BS.O         REFERENCES	B-38
                                                                          DRAFT

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                                           iii
                                 LIST OF TABLES
                                                                                Page No.

Table Bl.      Summary of Data Requirements for Prediction of the Quality of Effluent from
              Confined Dredged Material Disposal Areas.                                 B-13

Table B2.      Recommended Resuspension Factors for Various Ponded Areas and Depths.      B-20

Table B3.      Observed Flocculent Settling Data.                                         B-31

Table B4.      Percentage  of Initial Concentration and Suspended Solids Concentrations vs.
              Time, Assumed Depth of Influence of 2 ft.                                  B-33
                                                                          DRAFT

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                                            IV
                                 LIST OF FIGURES
                                                                                 Page No.

Figure Bl.     Schematic of Supernatant Water Interaction in an Active Confined Disposal
              Facility.                                                                 B-2

Figure B2.     Flowchart  Illustrating Approach for Evaluating Potential Effluent  Impacts
              from Confined Dredged Material Disposal Areas.                              B-9

Figure B3a.    Specifications for Settling Column and Plan for Sedimentation Column.         B-16

Figure B3b.    Plans for Top and Bottom Columns.                                        B-17

Figure B4.     Concentration Profile Diagram.                                            B-31

Figure B5.     Plot of Supernatant Suspended Solids Concentration vs. Time from  Column
              Settling Tests.                                                           B-32
                                                                           DRAFT

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	B-l	

APPENDIX B -       GUIDANCE FOR EVALUATION OF EFFLUENT DISCHARGES FROM
                      CONFINED DISPOSAL FACILITIES

Bl.O          INTRODUCTION

Bl.l          Background

Dredged material may be placed in diked disposal areas sometimes called confined disposal facilities
(CDFs).  CDFs  may  be considered as an  alternative  for  contaminated  dredged material that  is
unsuitable for disposal in open water.  Possible contaminant migration pathways for confined disposal
facilities include effluent discharges to surface water during filling operations, surface runoff due  to
precipitation, leachate into groundwater, volatilization to the atmosphere, and direct uptake by plants
and animals.  Subsequent cycling through food webs to animal  populations living in close association
with the dredged material should also  be considered.  Each pathway may have its own standards and
criteria defined  by the  water  quality certification  or other applicable  laws  and regulations.   If
standards or criteria  are not met, management options  may be considered including operational
modifications, treatment or containment options such as covers or liners.

This appendix provides  technical  guidance  for evaluation of  the effluent pathway.   Guidance for
evaluation of other pathways  and for management actions and control measures for CDFs is found  in
USACE/EPA (1992).

Dredged material may be placed in CDFs in  several ways.  The most common method of filling is by
direct  hydraulic pipeline from cutterhead dredges.  Pumpout operations from hopper dredges  or
hydraulic reslurry from barges  results in intermittent hydraulic filling.  Direct mechanical placement
of dredged material from barges (or possibly from trucks) can be done with equipment located at the
CDF.  All of these operations result in some sort of effluent discharge, defined for purposes of this
manual as that material discharged  directly to receiving waters during the filling operation (this would
include water discharged directly over  weir structures or through filter cells or retaining dikes).

A schematic of an active hydraulically filled CDF is shown in Figure Bl. Dredged material hydra-
ulically placed in a confined disposal area settles, resulting in a thickened deposit of material overlaid
by  a clarified supernatant.  The supernatant waters are discharged from  the site as  effluent during
active  dredging  operations.   The  effluent may contain  both dissolved contaminants  and suspended
colloidal particles with associated (adsorbed  or held by ion exchange) contaminants.  A large portion
of the total contaminant concentration is particle associated.
                                                                               DRAFT

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                                                B-2
  INCOMING
  DREDGED MATERIAL
                                                                          WEIR
                        TURBULENT MIXING
                        AND OXYGENATION
                               fC>« I «P»l/^J^  -  •  ,  -r.ii ; -- ^ .
                               OF FINE  V  ^ ',•'•_•• ^\ • \ '-.  - _  •
                              ./C£.ES__,-   fJ • •• ;§  '%.'if)\ ^\ .' .  '
                            • ^	~ "FLUX OF 'DISSOLVED FRACTION > <1} ' '  -
                           .^. .         PLUS P»BT/C1.ES ;  ) ; • •, !-"-{- -
                                                        SEDIMENTATION OF '
                                                     .SUPERNATANT PARTICLES
Figure Bl.     Schematic of Supernatant Water Interaction in an Active Confined Disposal Facility.

Supernatant waters from confined disposal sites are discharged after a retention  time of up  to several
days.  Furthermore,  actual withdrawal of the supernatant is governed by the hydraulic characteristics
of the ponded area and the discharge weir.  Several factors influence the concentration of suspended
particles present in  supernatant waters.  Fine particles become suspended in the disposal area water
column at the point of entry due to turbulence and  mixing.  The suspended particles are partially
removed from the water column by sedimentation.  However, particle  concentrations  may be  main-
tained by flow of water through the slurry  mass during settling.  Wind  and/or surface wave  action
may also resuspend  additional particles.  All solids cannot be  retained during  the  disposal process,
and associated contaminants are transported in dissolved form and with the particles in the effluent.
                                                                                   DRAFT

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	B-3	

B1.2          Purpose and Scope

The purpose of this appendix is to describe procedures for evaluation of effluent discharges from
CDFs.  The procedures provide an estimation of potential contaminant release and/or biological effect
under laboratory-simulated  confined disposal conditions and consider the sedimentation behavior of
dredged material, the retention tune of the proposed containment area, and the physicochemical envi-
ronment in ponded water during active disposal into the containment area.
B1.3          Regulatory Considerations

The quality of effluent discharged from these sites is an environmental concern and is regulated as a
discharge under Section  404 of the Clean Water Act.   In addition, Section 401 provides the States a
certification role as to project compliance with  applicable State water  quality  standards;  effluent
standards may be set as a condition of the certification.

The discharge of effluent from a CDF is defined as a dredged material discharge in 33 CFR 323.2 (d)
and 40 CFR 232.2 (e):

       The term "discharge  of dredged material" means any addition of dredged material into
       waters  of the  United States.   The term includes,  without limitation,  the addition of
       dredged material  to  a  specified discharge site located in waters  of the United States
       and the runoff or overflow from a contained land or water disposal area.

Section 230.10 (c)(l) of the Guidelines states that no discharge of dredged material shall be permitted
which will result in significant adverse impacts on  municipal water supplies.  Section 230.50 (a)
defines municipal water supplies as surface or groundwater directed to the intake of any municipal or
private water supply system.  Therefore, the potential impacts of leachate into groundwater must also
be considered.

The USAGE  issues  individual permits  and general permits.   Individual permits  are  issued  on a
project-by-project basis after the Guidelines compliance and public interest determinations are  made
for the specific project  at issue.   General permits,  on the other hand,  are issued for  classes of
activities  and/or activities conducted  in certain classes of  waters  of the U.S. after the US ACE
conducts the Guidelines  compliance and public interest reviews and determines  that issuance of the
general permit  will not result in more than minimal adverse impacts to the aquatic environment from
either a site-specific or cumulative standpoint. General  permits require little or no reporting, analysis,
or paperwork, compared to individual  permits.
                                                                                 DRAFT

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	B-4	

There are three types of general permits issued by the USAGE, nationwide permits, regional general
permits and programmatic general permits.  Nationwide permits are issued by the Chief of Engineers
and  apply nationwide.   Regional permits  are issued by district and  division engineers  and  are
applicable  on  district  or State-wide basis.   Programmatic permits  are issued (by  the  Chief  of
Engineers, as well as district and division engineers) to other federal, State or local agencies with the
intention of  providing  the appropriate level of environmental protection and avoiding unnecessary
duplication of effort with  the agency regulatory activities at issue.

There  are  currently four nationwide permits that pertain to  dredging and the discharge of dredged
material. One  authorizes  the discharge and  return water from  confined  disposal areas  (provided the
associated dredging is authorized pursuant to Section 10 of the River and Harbor Act of 1899); two
other nationwide permits  authorize the dredging and discharge, respectively, of up to 25 cubic yards
of material; and a fourth  authorizes maintenance dredging of existing marina basins (provided that the
dredged material is deposited on uplands; return water from a confined disposal area  requires separate
authorization pursuant  to  section 404 of  the Clean  Water Act).  As stated in the  preamble to  the
nationwide permit regulations (FR56, 226, November 22, 1991), the USAGE depends on its districts'
knowledge of potentially contaminated areas  and on the discretionary authority of district and division
engineers  to develop  special  conditions and/or require individual permits  where  contaminated
sediments are present.  General permits are not intended to apply to projects involving the dredging
or the discharge of contaminated materials.
B1.4          Applicability

B 1.4.1         Hydraulic Filling

The techniques for  evaluation of  effluent discharges described in this appendix  are specifically
designed  for  the  case of hydraulic placement  of material into CDFs with the effluent  discharge
occurring from an outlet pipe or weir structure or structures.  Hydraulic placement can be in the form
of direct pipeline  inflow from cutterhead or similar hydraulic suction dredges, intermittent hydraulic
placement from hopper dredge pumpout operations, or  intermittent hydraulic placement by reslurrying
material from barges (which  may have  been  filled by  mechanical  dredges).   Such  placement
operations would normally have an effluent discharge flowrate roughly equal to that of the inflow.
                                                                                  DRAFT

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	B-5	

Bl.4.2         Flow Through Dikes

Some CDFs may be designed to allow flow of effluent water through  filter cells or permeable dike
sections.  The techniques described in this appendix may be applied to this case, but the influence of
the filter media  in adsorption of contaminants from  the effluent  discharge  should be considered
(Krizek et al., 1976).


B 1.4.3         Mechanical Filling

Dredged material may be placed in some CDFs by direct mechanical means  such as rehandling from
barges or by truck.  Although such filling operations normally  involve handling relatively  little free
water, there may still be an effluent discharge.  Also, there may be ponded water in the CDF before
filling begins,  especially  for CDFs constructed in water.  For the case of mechanical filling, the
effluent discharge involves the free water which  is released during the mechanical placement operation
or the existing pond water which  is displaced by the operation.  No laboratory-developed and field-
verified  techniques now exist  for  the  case of  direct mechanical  placement of materials in CDFs,
however the procedures described  here  may  be used in the  interim for  the case of mechanical
placement and are considered conservative for such evaluations.


Bl.4.4         Surface Runoff and Leachate

Long-term geochemical changes may occur following disposal, site dewatering,  and  subsequent drying
of the  dredged  material.   The  quality of  the surface  runoff  or leachate   to  surface  water or
groundwater from disposal sites after these long-term changes occur may  be  markedly different from
that of the effluent discharged during active disposal.  The techniques described  in this appendix apply
only to  conditions  during active filling of the site and do  not account  for long-term geochemical
changes.  Therefore, they should not be used to evaluate the quality  of surface  runoff or leachate.  In
accordance with  33 CFR 336.1 (b)(8)  and Corps  Regulatory  Guidance Letter 87-8,  the  technical
procedures contained in USACE/EPA (1992) should be used as a guide for developing the appropriate
tests and evaluating surface runoff and leachate and possible management options.
                                                                                DRAFT

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	B-6	

B2.0          OVERVIEW OF EVALUATIONS  FOR EFFLUENT DISCHARGES

The discharge of effluent from CDFs has the potential for water column effects only.  Any solids in
the effluent would be dispersed and mixed.  Because CDFs are designed to retain virtually all of the
solid fraction of dredged material,  the evaluation of benthic effects is usually not applicable.

The evaluation  of  water column  effects resulting from  effluent discharges  uses  a tiered approach
generally patterned after that for  discharges of material  into open water.  General guidance in the
main body of this manual [pertaining  to  Tier  I evaluations (Sections 4.0  and  4.1), selection  of
contaminants  of concern  (Section 4.2), sample  collection and  preservation  (Section 8),  analytical
procedures  (Section 9)  and  general procedures  for  toxicity tests  (Section 10)]  is applicable for
evaluation of effluent discharges.
B2.1          Water Quality Standards

Section 401 of the CWA requires that all Federal permits and licenses, including those for effluent or
other discharges into waters of the United States, authorized pursuant to Section 404 of the CWA,
must be certified as complying with applicable State water quality standards (WQS). Violations of any
applicable State water quality standard apply at the edge of a State designated mixing zone.

The process for adoption of State  WQS is prescribed at 40 CFR  131. States must issue, condition,
deny,  or  waive a  Water  Quality  Certification for  activities  permitted  or  conducted  by USAGE,
certifying that no adverse water quality impacts will occur based on determinations of compliance
with applicable State WQS which  have been  adopted in accordance with the above regulation. State
water quality standards consist of designated uses,  narrative and numeric criteria designed to support
those uses, and anti-degradation provisions.
B2.2          Mixing Zones

The  evaluation  of  effluent discharges  must  consider the  effects  of mixing  and  dispersion.
Section 230.3(m) of the Guidelines defines the mixing zone:

        The term "mixing zone" means a limited volume of water serving as  a zone of initial
        dilution in  the  immediate vicinity  of a discharge point where receiving water quality
        may not meet  quality  standards or other requirements otherwise applicable  to  the
                                                                                 DRAFT

-------
	B-7	

       receiving water.  The mixing zone should be considered as a place where wastes and
       water mix and not as a place where effluents are treated.

Mixing zones are normally defined by the State regulatory agency as part  of the 401  water quality
certification. Detailed procedures for  evaluation of mixing zones for  CDF discharges are found in
Section B4.0.
B2.3          Basis of Evaluations

Chemical analyses are performed for contaminants that may be released from dredged material placed
in CDFs and  the  results are  compared  to water  quality standards for these contaminants  after
allowance for mixing.  This provides an indirect evaluation of potential biological impacts.  If water
quality  standards are met for all contaminants of concern, the material  discharged  as effluent in the
water column may also be evaluated for toxicity after mixing. Toxicity tests provide information on
the  toxicity  of contaminants not  included in the  water quality standards, and  indicate possible
interactive effects of multiple contaminants.  Bioaccumulation from the material discharged as effluent
in the water column is typically considered to be of minor concern due to the short exposure time and
low  exposure concentrations resulting from rapid dispersion and dilution.
B2.4          Contaminant Controls

If the testing  and associated  analysis  of the  effluent pathway indicates  applicable water  quality
standards will  not be met  after consideration of mixing, appropriate contaminant controls may be
considered to reduce impacts to acceptable levels.  Controls for effluent may include  modification of
the operation (e.g., use of a smaller dredge with reduced inflow rate, providing increased ponded area
and depth of the  CDF,  or relocation  of the  inflow and effluent  discharge points), treatment or
filtration of effluent to reduce the concentration of suspended solids and associated contaminants in the
effluent, and treatment of effluent to  remove dissolved contaminants. Additional  information  on
contaminant controls is found in USACE/EPA (1992).
B2.5          Tiered Approach

The tiered approach for evaluation of water column effects for effluent discharges from CDFs is
generally patterned  after that for  discharges of material into open water (Section 3.1).   The Tier I
evaluations  should  be  conducted  as  described  in Section 4.0.   Procedures  in this appendix for
                                                                                 DRAFT

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	B-8	

evaluation of effluent  discharges are  performed in Tiers II  and III.  A  flowchart  illustrating the
approach  for evaluating potential effluent impact is shown in Figure B2. Detailed descriptions of the
test procedures  are given in Section 63.0.    Tier IV evaluations for effluent discharges, if deemed
required,  would be performed considering the guidance in Section 11.3.

Tier II  evaluations for effluent discharges  consist of determinations of a screen  relative  to  WQS
compliance and  perhaps conduct of additional water column testing.  Water column testing should be
conducted only  if shown by the evaluation to be necessary.  Water column impacts are evaluated (if
necessary) by comparison of applicable water quality standards to the contaminant concentrations in
the effluent discharge after  consideration of mixing. Water column impact must also be evaluated by
toxicity testing in Tier III when there are contaminants of concern for which applicable WQS are not
available or where interactive effects are of concern..
B2.6          Tier II:  Water Quality Evaluations

B2.6.1         Screen Relative to WQS

The screen relative to  WQS  determines  the need  for additional  testing by  considering  the  bulk
concentration of contaminants in the dredged  material, the mixing at the disposal site, and applicable
water quality standards.  If the need for additional testing is not demonstrated,  the effluent discharge
complies  with  WQS. If additional testing  is needed, it is conducted according to  the guidance in
Section B3.0 as appropriate.

The screen involves a determination of whether the water quality standards,  after  consideration of
mixing, would be met if the bulk concentration of contaminants present in the sediment were to be
completely dissolved in the effluent water discharged from the disposal site.

The contaminant that would require the greatest dilution is determined by calculating the dilution that
would be required to meet the applicable water  quality  standard.  To determine the dilution (D) the
following equation is solved for each contaminant of concern:

                              D = [(C. x SS/1000) -

where          C.      =      concentration of the contaminant in the dredged material expressed as
                              micrograms  per kilogram (/ig/Kg), on a dry weight basis;
                SS     =      suspended solids  concentration in the effluent discharge expressed as
                              grams per liter (g/L);
                                                                                 DRAFT

-------
                                                            B-9
                                                     EVALUATE PROJECT DATA
                                                                            EFFLUENT TOXICITY
                      WQS SCREEN,
                         ASSUME
                TOTAL RELEASE OF SEDIMENT
                   CONTAMINANTS IN THE
                        EFFLUENT
                         (B2.S1)
                                                    MEASURE DISSOLVED
                                              CONCENTRATIONS OF CONTAMINANTS
                                                  OF CONCERN IN EFFLUENT
                                                         (101.2)
EXCEED WOS AFTER
                                                     MODEL DISSOLVED
                                              CONCENTRATIONS OF CONTAMINANTS
                                                  OF CONCERN IN EFFLUENT
                                                          (10.1.2)
                                                    EXCEED WOS AFTER
                                MEASURE TOXJCJTY
                                OF DM SUSPENSION
                                     (11.1)
                                 TOXICITY > 10%
                                 DIFFERENCE AND
                             SIGNIFICANTLY DIFFERENT
                                 THAN DILUENT
                                    WATER?
                                   MODEL DM
                               SUSPENDED PHASE
                               IN WATER COLUMN
                                    (11.1.7)
            DM NOT
           PREDICTED
          TO RESULT IN
            ACUTE
         WATER-COLUMN
            TOXICITY
               MODELED
            CONCENTRATION
         EXCEEDS 0.01 OF LC KjOfl
           ECjo AFTER INITIAL
                MIXING?
                (11.1.8)
    DM
 PREDICTED
 TO RESULT IN
WATER-COLUMN
   TOXICITY
                                                                                                      TIER  H
                                                                                  TIER  ILT
                     KEY TO NOMENCLATURE

             DM  DREDGED MATERIAL

            WOS  WATER QUALITY STANDARDS

            LCjo  LETHAL CONCENTRATION TO 50% OF
                 TEST ORGANISMS. EQUAL TO
                 ACUTE TOXKXTY CONCENTRATION

            ECjo  EFFECTS CONCENTRATION: EQUIVALENT
                 TO LC50 FOR NON-LETHAL ACUTE EFFECTS
Figure 62.      Flowchart  Illustrating  Approach  for  Evaluating  Potential  Effluent  Impacts   from
                   Confined Dredged Material Disposal Areas.
                                                                                                        DRAFT

-------
	B-10	

               1000   =      conversion factor, g to Kg;
               Q,     =      WQS in micrograms per liter Otg/L); and
               Cj.     =      background concentration of  the  contaminant at the disposal  site in
                              micrograms per liter (/tg/L).

The mixing zone evaluation is then made for the contaminant that would require the greatest dilution.
If the concentration after mixing is below the applicable water quality standard, the effluent discharge
complies with WQS.  If this concentration exceeds  the applicable  water quality standard, additional
testing must be conducted according to the guidance in Sections B2.6.2 and B3.0.
B2.6.2        Testing for Evaluation of Effluent Water Quality

The Tier II water  column  evaluation considers concentrations of contaminants of concern released
from the dredged material (in contrast to bulk concentrations used in Section B2.6.1), after allowance
for  initial mixing,  compared with  applicable water  quality standards.   The  evaluation therefore
requires a prediction of the CDF effluent quality.

Depending on the basis of applicable water quality standards, the prediction of the quality of effluent
from CDFs must account for the dissolved concentration of contaminants and  may also consider that
fraction associated  with the released total suspended solids.   A modified elutriate procedure has been
developed for this  purpose (Palermo, 1986, 1988; Palermo and Thackston, 1988a and b).  This test
defines dissolved concentrations of contaminants  in milligrams per liter and contaminant fractions in
the  suspended solids  (SS)  in milligrams per kilogram SS  under quiescent  settling conditions and
considers the geochemical  changes occurring  in  the disposal area during  active disposal operations.
Refinements and extensions of column settling test procedures (Averett et al., 1988;  Montgomery et
al.,  1983; and Palermo and Thackston,  1988c) have also been developed to define the concentration
of SS in the effluent for  a  given operational condition (i.e., ponded area and depth, inflow rate, and
hydraulic efficiency).   Using  results from both of these analyses,  a prediction of the total  concen-
tration of contaminants in the effluent can be made.  If no standards exist for effluent total  suspended
solids, turbidity, or whole water concentrations, the column settling test need not be conducted.  If no
standards for whole  water contaminants exist,  the  modified elutriate need  only be analyzed for
dissolved concentrations.

Predicted contaminant concentrations based on the results of a modified elutriate test can be used with
applicable water quality  standards to determine if the discharge  is  in compliance with the standards
after consideration  of mixing.  To determine the dilution (D)  required to meet the standards, the
following equation is solved for each contaminant of concern:
                                                                                  DRAFT

-------
                                             B-n
                             D = (CL - C^,) /  (CU, - CJ
where  C^ =       concentration  of the  dissolved  contaminant  in  the  modified elutriate  in
                      micrograms per liter Gig/L). All other terms  are  as  previously defined  in
                      Section B2.6.1.

The mixing zone evaluation is then made -for the contaminant that would require the greatest dilution.
If the concentration  after mixing is below  the  applicable water  quality  standard,  the  discharge
complies with WQS. Otherwise, it does not.
B2.7          Tier III: Toxicity Evaluations

Tier III testing assesses the impacts of contaminants in the dredged material on appropriate sensitive
organisms to determine if there is potential for the dredged material to have an unacceptable adverse
impact.  The Tier III  assessment  methods  are  toxicity  tests,  which  use lethality  as  the  primary
endpoint because  the  importance of this  endpoint  is easily interpreted.   These  acute tests use
organisms representative of the water column at the disposal site.  The recommended procedures for
water column toxicity tests for evaluation of effluent discharges are conducted in generally the same
manner as those for discharges of material  into open water (Section 11.1).  The only exception  is that
the toxicity test medium is  prepared using a modified elutriate procedure.

The results of the water column  toxicity tests must be interpreted considering the effects of mixing.
If the concentration  of dissolved  plus suspended  contaminants, after allowance for mixing, does not
exceed 0.01 of the toxic (LC50 or EC50) concentration beyond the boundaries of the  mixing zone,
the discharge  is predicted not to be acutely toxic to water column organisms. If the concentration of
dissolved  plus suspended  contaminants,  after allowance for mixing,  exceeds  0.01  of the  toxic
concentration, the discharge is predicted to  be acutely toxic to water column organisms.
B3.0           TESTING PROCEDURES FOR EFFLUENT DISCHARGES

This section describes the data requirements, testing procedures, and evaluation techniques necessary
to predict effluent contaminant concentrations for the Tier II evaluation and to conduct water column
toxicity tests for the Tier III evaluation. Example calculations are presented as appropriate.

The predictive techniques can be applied  to evaluate the performance of existing sites and to design
new sites.  For existing sites, the technique can be used to predict the effluent quality for a given set
                                                                                DRAFT

-------
	B-12	

of anticipated operational conditions (known flow and containment area size).  In a similar manner,
the required operational conditions for a new site (size, geometry, maximum allowable dredge size,
etc.)  can be determined  to  meet a given  effluent quality requirement  by comparing the predicted
effluent quality for a variety of assumed operational conditions.  In either case evaluation of effluent
quality must be considered in conjunction with a sound design of the CDF for retention of suspended
solids and initial storage of the sediments to be dredged.
B3.1          Data Requirements

Data requirements   for  prediction  of  effluent quality include  those pertaining  to  operational
considerations  (i.e.,  CDF site characteristics and dredge characteristics) and those  pertaining to the
properties of the dredged material (i.e., contaminant release characteristics and sedimentation charac-
teristics).  Data relating to operational  considerations  are usually determined  by the disposal area
design and by  past experience in dredging and disposal  activities for the project under consideration
or for similar  projects.  Data  relating  to  the dredged  material characteristics  must be obtained by
sampling the sediments to be dredged and testing  them.  A  summary of the data  requirements for
effluent quality prediction is given in Table Bl.
B3.1.1        Disposal Area Design

When the quality of the effluent from a CDF is of concern, the design, operation, and management of
the site should be carefully controlled.  This includes aspects relating to both the volume required for
effective sedimentation and the  storage  capacity  of the site.  Procedures for such evaluations are
presented in Engineer Manual 1110-2-2-5027 (USAGE, 1987), and should be considered prior to the
prediction of the quality of the effluent for the project.  These design procedures will determine the
surface area and ponding depth required to achieve effective sedimentation, the required containment
volume for storage (including required  freeboard),  and the proper sizing of weir structures.  The
prediction of the quality of the effluent described in this appendix is an extension and refinement of
the design procedures.  A list of data items required from the design evaluation is shown in Table Bl.

The process described in Section 4.2 should identify which contaminants are of  concern and which
therefore should be considered for subsequent analysis in the modified elutriate testing.  The modified
elutriate tests and the column settling tests provide the remaining data required for prediction of the
quality of the effluent.
                                                                                  DRAFT

-------
	B-13	

Table Bl.      Summary  of Data Requirements  for Prediction  of the Quality of  Effluent  from
               Confined Dredged Material Disposal Areas.
      Data Required
Dredge inflow rate

Dredge inflow solids
concentration

Ponded area in disposal site

Average ponding depth in
disposal site and at die weir

Hydraulic efficiency factor
Effluent total suspended
solids concentration

Dissolved concentration of
contaminant in effluent

Fraction of contaminant in
the total suspended solids in
effluent
Smbol
       Source of Data
  A,

  IV


  HEF


  sseff
   SS
Project information; site design

Project information; site design


Project information; site design

Project information; site design
Dye tracer or theoretical
determination

Laboratory column settling tests
                     Modified elutriate tests
                     Modified elutriate tests
        This summary includes only those data required for effluent quality prediction.  It is assumed
        that the disposal area under consideration is designed for effective sedimentation and storage
        capacity.   Data  requirements for such design or  evaluation are found in EM 1110-2-5027
        (USAGE, 1987).
                                                                                DRAFT

-------
	B-14	

B3.1.2         Sampling Requirements

Samples of channel sediment and water from the dredging site are required for conducting modified
elutriate tests and column settling tests, and toxicity tests,  and for characterizing the sediment to be
dredged.  The level of effort, including number of sampling stations, quantity  of material, and any
schemes used for compositing samples, is  highly  project-specific.  If at all  possible, the sampling
operations required for sediment characterization (both physical and chemical), design and evaluation
of the disposal site, and conducting the modified elutriate tests or toxicity tests should be conducted
simultaneously to avoid duplication of effort.  Note that water from the dredging site is used in tests
for evaluation of effluent  discharges.  Dredging site water is used since the  effluent  discharge only
involves a  small fraction of  dredged material  solids and the fractionation of contaminants  to  the
dissolved phase will be influenced primarily by that water. Note that disposal site water samples must
also be taken and analyzed for evaluation of mixing.  The guidance in Section  8 should be used for
obtaining samples.
B3.2          Column Settling Tests

Settling tests are necessary to provide data for design or evaluation of disposal areas for retention of
suspended solids.  These tests are designed to define the settling behavior of a particular sediment and
to provide information concerning the volumes occupied by newly placed layers of dredged material.
For purposes of effluent water quality prediction, the column setting tests need only be performed if
the water quality standards for chemical contaminants are defined in terms of total concentrations or if
there are  water quality  standards  for  total suspended solids or turbidity.  If standards exist  for
turbidity, a sediment-specific correlation of suspended solids and turbidity must be developed.

Sedimentation of freshwater slurries of  concentration less than 100 g/L can generally be  characterized
as  flocculent settling.   As slurry  concentrations are  increased,  the sedimentation process may be
characterized as a zone settling process, in which a clearly defined interface is formed between the
clarified supernatant water and the more concentrated settled material.  Zone settling also occurs when
the sediment/water  salinity is approximately 3 ppt or greater.  Flocculent  settling  also describes the
behavior  of residual suspended solids  in the clarified supernatant water  above the sediment/water
interface   for  slurries  exhibiting  an  interface.    The  procedures  described below   define  the
sedimentation  of  suspended solids under  flocculent  settling  conditions  or above  the  settled
material/water  interface under  zone setting conditions.   The settling test procedures  consist  of
withdrawing  samples from  the settling  column at various depths  and  times  and  measuring  the
concentrations of suspended solids.
                                                                                   DRAFT

-------
	B-15	

B3.2.1         Apparatus

An 8-inch diameter settling column such as shown in Figure B3  is used.  The test column depth
should approximate the effective settling depth of the proposed disposal area. A practical limit on the
depth of the test is 6 ft.  The column should be at least 8 in. in diameter with interchangeable sections
and with sample ports at 1/2-ft or closer intervals.  The column should have provisions to bubble air
from the bottom to keep the slurry mixed during  the column filling period.
B3.2.2         Test Procedure

The following test procedure should be used:

Step  1.   Mix the sediment  slurry  to a suspended  solids concentration C  equal to the expected
concentration of the dredged  material influent Q .  The slurry should be mixed in a container with
sufficient volume to fill the test column.  Field studies indicate that for maintenance dredging of fine-
grained  material,  the disposal  concentration  will  average  about  ISO grams   per  liter.    This
concentration should be used in the test if better data are not available.

Step 2.   Pump or pour the slurry into the test column  using compressed air or mechanical agitation to
maintain a uniform concentration during the filling  period.  Any coarse material which settles to  the
bottom of the mixing  container during transfer of the slurry to the test column need not be added to
the column.

Step 3.   When the slurry is completely mixed in the column, cut off the compressed air or mechanical
agitation and immediately draw off samples at each  sample port  and determine their suspended solids
concentration.  Use the average of these values as  the initial  slurry concentration  at the start of  the
test.  The test is considered initiated when the first samples are drawn.

Step 4a.  If an interface has  not  formed during the first day, flocculent  settling is occurring in  the
entire slurry mass.   Allow the slurry to settle and  withdraw samples from each sampling  port at
regular  time intervals to  determine the suspended  solids concentrations.   Record  the water  surface
height and time at the start of the sampling period.  Analyze each sample for total suspended solids.
Substantial reductions of suspended  solids will occur  during the early part of the test, but reductions
will decrease with  longer retention times.  Therefore, the intervals can be  extended  as  the test pro-
gresses.  Recommended sampling intervals are 1, 2, 4, 6, 12,  24, 48 hours, etc., until the end of the
test.  As a rule, a 50-milliliter sample should be taken  from each  port.  Continue the test until either
                                                                                 DRAFT

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fluid above the  interface is less than 1 gram per liter or until the suspended solids concentrations  in
extracted samples shows no decrease.

Step 4b.   If an interface forms  the  first day, zone settling is occurring  in the slurry  below the
interface, and flocculent settling is occurring in the supernatant water.  For this case, samples should
be extracted from  all side ports above the falling interface.  The first of  these samples should be
extracted  immediately after  the interface has fallen  sufficiently below the uppermost port to allow
extraction  or sufficient sample can be withdrawn  from the surface without disturbing the interface.
This sample can usually be extracted within a few hours after the beginning of the test.  Record the
time of extraction,  water surface height, and port height for each port  sample taken and analyze each
sample for suspended solids.  As  the interface continues to fall, extract samples from all ports above
the interface at  regular time intervals.  As before, a suggested sequence of sampling intervals  would
be 1, 2, 4, 6, 12,  24, 48, 96 hours,  etc.   The samples  should continue to be taken until either the
suspended solids concentration of the extracted samples shows no decrease or for a maximum time  of
15 days.  For this case, the suspended solids in the samples should be  less than 1 gram per liter, and
filtration  will  be  required  to  determine the  concentrations.    The  data  should be expressed  in
milligrams per liter for these samples.  In reducing the data for this case, the concentration of the first
port sample taken above the falling interface is considered the initial concentration SS0.
B3.2.3        Data Analysis

A flocculent data analysis procedure,  as outlined in the following paragraphs,  is required.  Example
calculations are also shown in Section  B3.4.

Step  1.  Arrange the flocculent settling test data from the laboratory test as shown in Table B3 and
compute values of the  depth of sampling  below the fluid surface, z.  In computing the fractions  of
suspended solids remaining 4>, the highest concentration of the first port samples taken is considered
the initial concentration SS0.

Step 2.  Plot the values of # and  z  using the data from  the table as shown in Figure B4, forming a
concentration profile diagram. Concentration-depth profiles should be plotted for each time of sample
extraction.

Step  3.   Use  the  concentration  profile diagram to graphically determine percentages of suspended
solids removed R for the various time intervals for the anticipated ponding depth D^,, (the minimum
recommended ponding  depth is 2 feet).  This is done  by graphically determining the area to the right
                                                                                  DRAFT

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	B-19	
of each concentration-depth profile and its ratio to the total area above the depth D^.  The removal
percentage is:
                            R m Area to right of profile  /10Q)
                                        Total areal
Step 4.  Compute the percentage  P remaining as simply 100 minus the percentage removed, or:

                                         p =  100 - R


Step 5.  Compute values for suspended solids for each time of extraction as:

                                         SS, = P,(SSa)


Arrange the data as shown in Table B4.

Step 6.   Plot a relationship for suspended solids concentration versus time using the value for  each
time of extraction,  as shown in Figure BS.  An exponential or power  curve fitted through the data
points is recommended.

By repeating steps 4 through 6  for each  of several values of D^  ,  a  family  of curves showing
suspended solids remaining versus retention time for each of several assumed ponding depths may be
developed.  These curves may be used for prediction of  effluent suspended solids concentrations
under ideal, quiescent settling  conditions for any estimated ponding  depth  and field mean retention
time. Simply enter a curve with the estimated field mean retention time  Td, and select the value of
effluent suspended solids predicted by  the column test SScol.  Guidance for determination of the  field
mean retention time is given in Section B3.2.5.   Guidance for adjusting  the value derived from the
column  test for anticipated resuspension is given  in the Section B3.2.4.


B3.2.4         Determination of Effluent Suspended Solids Concentration

A prediction  of the concentration of  total suspended  solids  in the effluent  must  consider the
anticipated actual mean retention time in the disposal area and  must account  for possible resuspension
of settled material because of wind-generated turbulence.  The relationship  of supernatant  suspended
                                                                                DRAFT

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	B-20	

solids versus time developed from the column settling test is based on quiescent settling conditions
found  in  the  laboratory.   The anticipated  actual mean retention  time  in the disposal  area under
consideration  can  be  used  to  determine  a  predicted  suspended solids concentration from the
relationship. This predicted value can be considered a minimum value which could only be achieved
in the field if there were little or no turbulence or resuspension of settled material.   However, an
adjustment for anticipated resuspension is necessary for real conditions. The minimum expected value
and the value adjusted for resuspension would provide a range of anticipated suspended solids concen-
trations for use in predicting the total  concentrations of contaminants in the effluent.   The value
adjusted for anticipated resuspension is:
                                        SS0 = SScol x RF

where
        SS^ =  suspended solids  concentration of effluent considering  anticipated  resuspension, mg
        suspended solids/L of water

        SSed = suspended solids concentration of effluent as estimated from column settling tests, mg
        suspended solids/L of water

        RF  = resuspension factor  selected from Table B2

Table B2 summarizes  recommended resuspension factors based on comparisons of suspended solids
concentrations predicted from column settling tests and field data from a number of sites with varying
site conditions.  For dredged material slurries exhibiting flocculent settling behavior, the concentration
of particles  in the ponded water is on the order of 1 g/L or higher.  The resuspension resulting from
normal wind conditions will not significantly increase this concentration.  Therefore, an adjustment
for resuspension is not required for the flocculent settling case.
Table B2.      Recommended Resuspension Factors for Various Ponded Areas and Depths.
                                    Resuspension Factor for Anticipated Average Ponded Depth
     Anticipated Ponded Area             Less than 2 ft.                  2 ft. or Greater
  Less than 100 acres                           2.0                             1.5
  Greater than 100 acres                        2.5                             2.0
                                                                                 DRAFT

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	B-21	

B3.2.5         Determination of Field Mean Retention Time

Estimates of the  field mean retention  time for expected operational  conditions  are  required  for
selecting appropriate settling times in the modified  elutriate test and for determination of suspended
solids concentrations  in the  effluent.  Estimates of the retention time  must consider the hydraulic
efficiency of the disposal area, defined as the ratio  of mean retention time to theoretical  volumetric
retention tune.  Field  mean  retention time   T4  can  be estimated for a given flow  rate and ponding
conditions by applying a hydraulic efficiency correction factor (HECF) to the theoretical detention
time as follows:
                                                „
                                               (HECF)
where
        Ta = mean detention time, h
        T = theoretical detention time, h
        HECF  = hydraulic efficiency correction factor (HECF  >  1.0) defined as the inverse of the
        hydraulic efficiency

The theoretical detention time is calculated as follows:
                                     v
                                 T =     (12.1)  = _     (12.1)
where
        Vp = volume ponded, acre-ft
        Qi = average inflow rate, cfs
        Ap = area ponded, acres
        Dp = average depth of ponding, ft
        12. 1 = conversion factor, acre-ft/cfs to h

The hydraulic efficiency correction factor HECF can be estimated by several methods.  The most
accurate estimate  is that made from dye  tracer studies to determine  Td at the actual  site under
operational conditions at a previous time, with the conditions similar to those for the operation under
consideration.  This approach can be used only for existing sites.

Alternatively, the ratio T/T = 1/HECF can be estimated from the equation:
                                                                                DRAFT

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                                             B-22
                                     -0.9  1 -exp-0.3
where LAV  is the  length-to-width  ratio of the proposed basin.   The LAV ratio can be increased
greatly by the use of internal spur dikes, resulting in a higher hydraulic efficiency and  a lower
required  total area.   In the absence of dye  tracer data or values obtained from other theoretical
approaches, a value for HECF of 2.25 may be used based on field studies conducted at several sites
(Montgomery, 1983; Montgomery et al., 1983).
B3.3           Modified Elutriate Test Procedure

The modified  elutriate tests  should be conducted,  and appropriate chemical analyses should  be
performed, as soon as possible after sample collection.  The volume of elutriate  sample needed for
chemical  analyses will vary  depending upon the number  and types  of chemical  analyses  to  be
conducted. Both dissolved and total concentrations of contaminants may be determined.  The volume
required for each analysis, the number of parameters  measured, and the desired analytical replication
will influence the total elutriate sample volume required.  A 4 L cylinder is normally used for the
test, and the supernatant volume available for sample  extraction will vary from approximately 500 to
1,000 mL, depending  on the sediment properties, settling  times,  and initial  concentration of the
slurry.  It may be necessary to composite several  extracted sample volumes or to  use large diameter
cylinders to obtain the  total required volume.
B3.3.1         Apparatus

The following items are required:
       a.      Laboratory mixer, preferably with Teflon shaft and blades.
       b.      Several  4 L graduated cylinders.  Larger cylinders may be used if large  sample
               volumes are required for  analytical purposes.  Nalgene cylinders are acceptable for
               testing involving analysis of inorganic compounds such as metals and nutrients.  Glass
               cylinders are required for testing involving analysis of organic compounds.
       c.      Assorted glassware for sample extraction and handling.
       d.      Compressed air  source with deionized water trap and tubing for bubble aeration of
               slurry.
                                                                                DRAFT

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	B-23	

       e.      Vacuum or pressure filtration equipment, including vacuum pump or compressed air
               source and an appropriate filter holder capable of accommodating 47-,  105-, or 155-
               mm-diam filters.
       f.      Presoaked filters with a 0.45 fim pore-size diameter.
       g.      Plastic sample bottles, 500 mL capacity for storage of water and liquid phase samples
               for metal and nutrient analyses.
       h.      Wide-mouth, 1 gal capacity  glass jars with Teflon-lined screw-type lids for sample
               mixing. These jars should also be used for sample containers when samples are to be
               analyzed for pesticides.

Prior to use, all glassware, filtration equipment,  and filters should be thoroughly cleaned.  Wash all
glassware with detergent, rinse  five times with tap  water, place in  a clean 10 percent (or stronger)
HCl acid bath for a minimum of 4 h, rinse five times with tap water, and then rinse five times with
distilled or deionized water.  Soak filters for a minimum of 2 h  in a 5 M HCl bath, and then rinse
10 times with distilled water.  It is also a good practice  to discard the first 50  mL of water or liquid
phase filtered.
B3.3.2         Test Procedure

The step-by-step procedure for conducting the modified elutriate test is outlined below.

Step  1  - Slurry preparation.  The sediment and  water  from the proposed dredging  site should be
mixed to a concentration approximately equal to the expected average field inflow concentration.  If
estimates of the  average field inflow concentration cannot be made based  on past  data, a  slurry
concentration of 150 g/L (dry weight basis) should be used.  Predetermine the concentration  of the
well-mixed sediment in grams per liter (dry weight basis) by oven drying a small subsample of known
volume. Each  4 L cylinder to be filled will require a mixed slurry volume of 3-3/4 L. The volumes
of sediment and water  to be mixed for a 3-3/4 L slurry volume may be calculated using the following
expressions:
and
                                     ^ = 3.75 -V.

where
               = volume of sediment, in L
                                                                                DRAFT

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                                              B-24

       3.75 = volume of slurry for 4 L cylinder, L
             ~ desired concentration of slurry, g/L (dry weight basis)
               = predetermined concentration of sediment, g/L (dry weight basis)
             = volume of disposal site water, in L
Step 2 - Mixing.  Mix the 3-3/4 L of slurry by placing appropriate volumes of sediment and water
from the proposed dredging site in a 1 gal glass jar and mixing for 5 min with the laboratory mixer.
The slurry should be mixed to a uniform consistency, with no unmixed agglomerations of sediment.

Step 3 - Aeration.   The prepared slurry  must be aerated to ensure  that oxidizing conditions will be
present in the supernatant water during the subsequent settling phase.  Bubble aeration is therefore
used as a method of sample agitation.  Pour the mixed slurry into a 4 L graduated  cylinder.  Attach
glass tubing to the aeration source and insert the tubing to the bottom of the cylinder.  The tubing can
be held in place by insertion through  a predrilled No. 4 stopper placed in the  top of the  cylinder.
Compressed air should be passed through  a  deionized water  trap, through the tubing, and  bubbled
through the slurry.  The flow  rate should be adjusted to agitate the mixture vigorously for 1 h.

Step 4 - Settling.  Remove the tubing, and allow the aerated slurry to undergo quiescent settling for a
time period equal to the anticipated field mean retention time,  up to  a  maximum of 24 h. If the field
mean retention time is not known, allow settling for 24 h.  Guidance for estimating the field mean
retention is given in Section B3.2.S.

Step 5 -  Sample extraction.    After the appropriate period  of quiescent settling,  an interface will
usually be evident between the supernatant water, with a low concentration of suspended solids above,
and the more concentrated settled material below the interface.  Samples of the supernatant water
should be extracted from the cylinder at a point midway between the water surface and interface using
syringe and tubing.  Care should be taken not to resuspend the settled material.

Step 6 - Sample preservation  and analyses.  The sample  should be analyzed as soon as possible after
extraction.   Total  suspended   solids  in  milligrams per  liter dissolved  and, if  required,  total
concentrations of desired  analytes in milligrams  per liter should be  determined.   (If water quality
standards for chemical contaminants  are in terms of dissolved concentrations,  the total concentration
of  contaminants and total  suspended solids in the modified  elutriate need not  be determined.)  If
required,  the fraction of analytes in the total suspended solids  in milligrams per kilogram SS can then
be  calculated  for appropriate  analytes.  Filtration using 0.4S  pm  filters should be  used to obtain
subsamples for  analysis of dissolved concentrations.   Samples to be analyzed for dissolved pesticides
or polychlorinated  biphenyls  (PCBs) must  be free of particles but should not be filtered due to the
tendency  for these materials  to adsorb on the filter.  However, paniculate matter can be removed
                                                                                  DRAFT

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	B-25	

before analysis by high-speed centrifugation at 10,000 times gravity using Teflon, glass, or aluminum
centrifuge tubes (Fulk et al., 1915).  The total suspended solids concentration can also be determined
by filtration (0.45 /un).  The fraction of analytes in the total suspended solids may be calculated  in
terms of milligrams per kilogram SS as follows:
                                   r, = (i x 10") ^L_
where
        FSJ = fraction of analyte in the total suspended solids, mg analyte/Kg of suspended solids
        Qrt.,  = total concentration, mg analyte/L of sample
        C&. = dissolved concentration mg, analyte/L of sample
        SS = total suspended solids concentration, mg solids/L of samples
B3.3.3        Chemical Analyses

Chemical analyses of the modified elutriate samples should be performed according to the guidance in
Section 9.
B3.3.4        Dissolved Concentrations of Contaminants

The dissolved concentrations of chemical  contaminants in the modified  elutriate are compared with
water quality standards after consideration of mixing (as described in Section 5.1.2) if such standards
are defined in terms of dissolved concentrations.
B3.3.5         Calculation of Total Concentrations of Contaminants

The modified elutriate test procedure estimates dissolved contaminant concentrations in milligrams per
liter and fractions of contaminants in the total suspended solids in milligrams per kilogram SS under
quiescent settling conditions  and accounts for geochemical changes  occurring in  the disposal area
during active disposal operations.  Using these test results, the total  contaminant concentration in
milligrams per liter in the effluent may be estimated as:
                                                                                 DRAFT

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                                             B-26
                                        C-, = C*
                                      tout     dbv
                                                 (1 x 10«)

where
                      estimated total concentration in effluent, mg analyte/L of water
       C&. =        dissolved concentration determined by modified  elutriate tests, mg analyte/L
                      of sample
       FM =         fraction  of  analyte in the total  suspended solids calculated  from modified
                      elutriate  results, mg analyte/Kg of suspended solids
       SS^ =        suspended  solids  concentration  of effluent  estimated from  evaluation  of
                      sedimentation performance, mg suspended solids/L of water
       (1 x  10") =   conversion factor, mg/mg to mg/kg

The acceptability of the proposed confined disposal operation can then be evaluated by comparing the
predicted total contaminant concentrations for the various contaminants with applicable water quality
standards, considering an appropriate mixing zone.
B3.4          Water Column Toxicity Test Procedure

The procedures for performing  toxicity  tests to evaluate  water  column effects of effluent discharges
from CDFs are generally  the same as those for  evaluation of dredged material  discharges in open
water (see Section S.I).  However, the preparation of the dredged material (dissolved plus suspended
contaminants) should done using the modified elutriate procedure as described below.

The volume required for each analysis, the number of parameters measured, and the desired analytical
replication will influence the total elutriate sample volume required.  A 4 L cylinder is normally used
for the test, and the supernatant volume  available  for sample extraction will vary from approximately
500 to 1,000 mL, depending on the sediment properties, settling times, and initial concentration of the
slurry.  It may be necessary to  composite several extracted sample volumes or to use large diameter
cylinders to obtain the total required volume.
B3.4.1         Apparatus

The following items are required:

        a.      Laboratory mixer, preferably with Teflon shaft and blades.
                                                                                 DRAFT

-------
	B-27	

       b.      Several 4 L graduated cylinders.   Larger cylinders  may be used if  large sample
               volumes  are required for analytical  purposes.   Nalgene cylinders  are acceptable for
               testing involving analysis of inorganic compounds such as metals and nutrients.  Glass
               cylinders are required for testing involving analysis of organic compounds.
       c.      Assorted glassware for sample extraction and handling.
       d.      Compressed air source with deionized  water trap and tubing for bubble  aeration of
               slurry.
       e.      Wide-mouth, 1-gal capacity glass jars  with Teflon-lined screw-type lids  for sample
               mixing.  These jars should also be used for sample containers when samples are to be
               analyzed for pesticides.

Prior to use, all glassware should be thoroughly cleaned.  Wash all glassware with detergent, rinse
five times with tap water, place in a clean bath for a minimum of 4 h, rinse five times with tap water,
and then rinse five times with distilled or deionized water.
B3.4.2         Test Procedure

The step-by-step procedure  for  conducting the modified elutriate test for use  in  toxicity tests is
outlined below.

Step 1  - Slurry preparation.   The sediment  and water from  the  proposed dredging site should be
mixed to a concentration approximately equal to the expected  average field inflow concentration.  If
estimates  of the average field inflow  concentration cannot be made based on  past data,  a slurry
concentration of 150 g/L (dry weight basis) should be used.  Predetermine the concentration of the
well-mixed sediment in grams per liter (dry weight basis) by  oven drying a small subsample of known
volume. Each  4 L cylinder to be filled will require a mixed slurry volume of 3-3/4 L.  The volumes
of sediment and water to be mixed for a 3-3/4 L slurry volume may be calculated using the following
expressions:
and

                                      v*atr = 3-75  -

where
       V^dimoit = volume of sediment, in L
                                                                                 DRAFT

-------
                                              B-28

       3.75 = volume of slurry for 4 L cylinder, L
             = desired concentration of slurry, g/L (dry weight basis)
               = predetermined concentration of sediment, g/L (dry weight basis)
              = volume of dredging site water, in L
Step 2 - Mixing.  Mix the 3-3/4 L of slurry by placing appropriate volumes of sediment and water
from the proposed dredging site  in a 1-gal glass jar and mixing for 5 min with the laboratory mixer.
The slurry should be mixed to a uniform consistency, with no unmixed agglomerations of sediment.

Step 3 - Aeration.   The prepared slurry  must be aerated to ensure that oxidizing conditions will be
present hi the supernatant water during the subsequent settling phase.  Bubble aeration is therefore
used as a method of sample agitation.  Pour the mixed slurry into a 4 L graduated cylinder.  Attach
glass tubing to the aeration source and insert the tubing to the bottom of the cylinder.  The tubing can
be held in place by insertion through  a predrilled No. 4 stopper placed  in  the top of the cylinder.
Compressed  air should be passed through  a deionized water  trap, through the tubing,  and bubbled
through the slurry.  The flow rate should be adjusted to agitate the mixture vigorously for 1 h.

Step 4 - Settling.  Remove the tubing, and allow the aerated slurry to undergo quiescent settling for a
time period equal to the anticipated field mean retention time,  up  to a maximum of 24 h.  If the field
mean retention time is not known, allow settling for 24 h.   Guidance  for estimating the field mean
retention is given in Section B3.2.S.

Step 5 -  Sample extraction.   After the appropriate  period  of quiescent  settling, an interface will
usually be evident between the supernatant water, with a low concentration of suspended solids above,
and the more concentrated settled material below the interface.

The liquid plus  the  material  remaining  in suspension  after  the  settling period  represents  the
100 percent  liquid plus  suspended paniculate phase.   Carefully siphon the  supernatant, without
disturbing the settled  material, and immediately  use it  for toxicity testing.  With some very fine-
grained dredged  materials, it may  be necessary to centrifuge  the supernatant for  a short time.  The
suspension should be clear enough at the first observation time for the  organisms  to be visible. The
general guidance in Section 10 should be followed in performing the toxicity tests.
                                                                                 DRAFT

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	B-29	

B4.0           EXAMPLE CALCULATIONS

B4.1           Example 1:  Evaluation of Effluent Water Quality For an Existing Disposal Area

This example illustrates the evaluation of a proposed effluent discharge for an existing CDF in which
effluent standards exist for dissolved contaminants and total suspended solids.


B4.1.1         Project Information

Dredged material from a maintenance project will be placed in an existing disposal site.  The ponded
area will be approximately 35 acres.   The  design  indicated  that the surface area  is adequate for
sedimentation if a minimum ponding depth of 2 ft is maintained.  The dredging equipment and pump-
ing conditions anticipated will result in a flow rate of approximately  30 cfs.  A dye tracer  test was
previously run  at this disposal site under similar operational conditions, and the field mean retention
time was 20 h.  Previous sampling  of inflow from the dredge pipe under similar conditions indicated
that the influent solids concentration was approximately ISO g/L, which is considered a conservative
maximum.

The quality  of effluent must be predicted and  compared with applicable water quality standards so that
the acceptability of the proposed discharge may be evaluated.  A field evaluation of dispersion at the
disposal  site determined that a dilution factor of 38 would  occur in the mixing zone.  For purposes of
this example, copper is  the parameter requiring the greatest dilution and will be used to illustrate the
calculations. The water quality standard for dissolved copper at the perimeter of the mixing zone was
set at 0.004 mg/L, while that for total suspended solids was set at 50 mg/L.
B4.1.2         Modified Elutriate Testing

Modified elutriate tests were conducted on  samples  of sediment and disposal site water from  three
stations at the site. The modified elutriate tests were run at the anticipated influent concentration, in
this case 150 g/L. Sediment samples for each sampling station to be tested were homogenized, and a
sediment concentration of 450 g/L was determined by oven drying a sample of known volume.   The
volumes of sediment and water mixed  for this sample for a 3-3/4  L slurry volume were determined
as:
                                                                               DRAFT

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                                             B-30
and
                              =  3.75 - V_.	 •= 3.75  - 1.25  =  2.50 L
The modified elutriate tests were completed with the retention time used  in the tests equal to the
anticipated field mean retention time of 20 h.  Samples were extracted for the replicate tests  and
analyzed for dissolved concentrations of desired parameters.  The mean concentration of dissolved
copper was 0.06 mg/L.
B4.1.3         Column Settling Tests

A column setting test  was required because of the  water quality standard for suspended  solids.
Samples  from all stations were homogenized into a composite for the  column settling test.  The test
used for  prediction of effluent suspended solids was run at a slurry concentration of ISO g/L, equal to
the anticipated influent  slurry concentration.  The interface was formed early in the  test.  Samples
were  extracted  from settling  column  ports at  3, 7,  14, 24,  and  48 h.   Data for the  solids
concentrations and for various depths and extraction times are shown in Table B3.

The concentration-depth profile  diagram  was  then constructed from the data, and is  shown in
Figure B4.   Ratios of  suspended solids  removed as  a  function of time  were  then  determined
graphically using the step-by-step procedure described previously.  Since an interface formed  in the
test, the slurry  mass  was  undergoing zone settling.   Therefore,  the  initial  supernatant  solids
concentration  SS0  was assumed to be the highest concentration of the first samples taken, 169 mg/L.
The concentration-depth  profile  diagram  was   therefore  constructed  using 169 mg/L  as   #  =
100 percent.  The lower horizontal boundaries for the area determinations corresponded to a range of
assumed  depths of withdrawal  influence at the outlet weir,  in this  case 1,  2,  and 3 ft.  An example
calculation of the removal  ratio  for the concentration-depth profile  at   t = 14 h and  a  depth of
influence of 2 ft is:
                 „  _  Area to right of the profile  _  Area 1230* - n  73
                  14              Total area               Area 1240
*  Areas are designated by circled numbers in Figure B5.  The areas were determined by planimeter.
                                                                                DRAFT

-------
                                           B-31
Table 63.     Observed Flocculent Settling Data.
    Sample Extraction
         Time
          t(h)
Depth of Sample
   Extraction
     z(ft)
Suspended Solids
   SS (mg/L)
Fraction of Initial SS
     $  (percent)
3
3
7
7
14
14
14
24
24
24
48
48
48
0.2
1.0
1.0
2.0
1.0
2.0
3.0
1.0
2.0
3.0
1.0
2.0
3.0
93
169
100
105
45
43
50
19
18
20
15
7
14
55
100
59
62
27
25
30
11
11
12
9
4
8
                           PERCENT OF INITIAL CONCENTRATION, %
                            30      40       50      60
                                                            100
                                                                    LEGEND

                                                                  D T=3 MRS

                                                                  O T = 7 MRS

                                                                  V T= 14 MRS
                                                                     T = 24HRS

                                                                      = 48HRS
             •~-T=24HRS


             -T=48HRS
Figure B4.     Concentration Profile Diagram.
                                                                            DRAFT

-------
	B-32	

The portion remaining at t =  14 h is:

                              P14  - 1 - *M  « 1 - 0.78 = 0.22
The value for the suspended solids remaining is:
                          SSU - PM (SS.) - 0.22  (169) - 37 »g/L
Values at other times  were determined  in  a similar manner.  The summary data are shown  hi
Table B4.  Similar calculations for other assumed ponding depths were made.  Curves were fitted  to
the data for total suspended solids versus retention time for depths of influence of 1, 2, and 3 ft and
are shown in Figure B5.
                                          LEGEND

                                           DEPTH OF INFLUENCE = 1.0 FT
                                           DEPTH OF INFLUENCE = 2.0 FT
                                           DEPTH OF INFLUENCE = 3.0 FT
                                EXAMPLE 1:

                                FIELD MEAN RETENTION = 20 HRS; SSCOL = 24 MG/L
                                          XAMPLE 2: SSCOL = 12 MG/L
                                         FIELD MEAN RETENTION = 36 HRS
                                         20            30

                                       RETENTION TIME, HRS
Figure BS.    Plot of Supernatant Suspended Solids Concentration vs. Time from Column Settling
              Tests.
                                                                            DRAFT

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	B-33	

Table B4.      Percentage of Initial Concentration and  Suspended Solids Concentrations vs. Time,
               Assumed Depth of Influence of 2 ft.
    Sample Extraction
          Time           Removal Percentage,    Remaining Percentage,      Suspended Solids
          t (h)                     R,                      P,                  SS (mg/L)
3 14
7 47
14 78
24 90
48 94
86
53
22
10
6
145
90
37
17
10
B4.1.4         Prediction of Effluent Suspended Solids Concentration

A value for the estimated effluent suspended solids can be determined for quiescent settling conditions
using the column test relationship.  In this case, the field mean retention time of 20 h corresponds to
a suspended solids concentration of 24 mg/L,  as shown in Figure B5.  This value should be adjusted
for anticipated resuspension using the factors shown in Table B2.  In this case, for a surface area less
than 100 acres  and an assumed average ponding depth of 2 ft,  the resuspension factor is  1.5.  The
predicted total suspended solids concentration in the effluent is calculated as :

                         SSeff = SScal x RF = 24  mg/L x  1.5 =36 mg/L
The acceptability of the discharge for suspended solids can be evaluated by comparing the estimated
effluent concentration with the water quality standard, considering the appropriate mixing zone.  For
suspended solids, the estimated concentration of 36 mg/L is less than the  water quality standard of
50 mg/L, therefore the discharge is acceptable for suspended solids prior to considering mixing.
B4.1.5         Prediction of Contaminant Concentrations

The acceptability of  the  proposed discharge for  contaminants  can be evaluated by  comparing  the
estimated effluent concentrations with applicable water quality standards, considering an appropriate
mixing  zone.   For  a mixing  zone  dilution of 38 and  a copper  standard  of 0.004 mg/L,  the
concentration of copper at the point of discharge must be less than 0.15 mg/L. The estimated concen-
tration of 0.06 mg/L from the modified elutriate test at the point of discharge is  less than the limiting
value of 0.15 mg/L. The discharge would therefore be acceptable.
                                                                                DRAFT

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	B-34	

B4.2           Example 2:   Determination  of Disposal Area Requirements  to Meet a  Given
               Effluent Quality Standard

This example illustrates  the  evaluation of a proposed effluent discharge for a new CDF in which
effluent standards are defined in terms of total  concentrations of contaminants. The required retention
time of the new CDF to meet the standards is determined.
B4.2.1         Project Information

A disposal area is planned for  contaminated  sediment  from a small maintenance dredging project.
Dredging equipment traditionally  used in the project area is capable of flow rates  up to IS cfs.
Available real  estate in the project vicinity is scarce, with the maximum available area limited  to
60 acres.  The disposal area required to meet  applicable water quality standards must be determined.
The CDF design indicated that a minimum ponded surface area of 20 acres was required for effective
sedimentation,  assuming a flow rate of 15 cfs and  an assumed minimum ponding depth of 2 ft.   A
mixing  evaluation was conducted using a computer model and a  dilution factor of 20  was estimated
for the  allowable mixing zone.  As for Example 1, copper  is the contaminant requiring the greatest
dilution.  The water quality  standard for copper  at the perimeter  of the mixing zone  was set  at
0.0029  mg/L (total copper).
B4.2.2         Modified Elutriate Testing

Modified elutriate tests were conducted and calculations were made as described for Example 1.  For
this example, the anticipated mean field retention time is not known beforehand, so  the maximum
laboratory retention of 24 h should be used for the tests.  Since the inflow concentration is not known
beforehand, the tests should  be run at a slurry  concentration of 150 g/L which is considered to be a
conservative maximum.   Results for the replicate tests  for  this example  are 0.006 mg/L for  the
concentration of dissolved copper and  2888  mg/Kg SS for the fraction  of copper in  the total
suspended solids.
B4.2.3        Column Settling Tests

Column settling tests were performed, and the resulting concentration-depth profile was developed as
was illustrated in Example 1.   The  column tests were  run at  a concentration of 150 g/L for this
                                                                                DRAFT

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	B-35	

example.  For simplicity, the test results from column tests used in the first example will also be used
in this example (see Figures B4 and BS).

B4.2.4        Determination of Allowed Effluent Suspended Solids Concentrations

Since this example requires determination of the disposal site characteristics necessary to meet a given
water  quality standard,  the calculations would proceed in a manner similar to Example 1,  but in
reverse sequence.  The concentration  of effluent suspended solids  required to meet water quality
standards  must first be determined.  For total copper, the standard (after consideration of mixing) is
0.0029 mg/L.  For a dilution of 20, the concentration of copper at the point of discharge must be less
than O.OS8 mg/L.  The suspended solids concentration required to meet this standard is:
                                    c    - c   +   FSS
                                    C    ~ C
or transposed,
            SS# =  (1 * 1Q6)  (C^ - C^)  = iflfl.1(? (0.058 - 0.006) =  18 mg/L
                       f cc                    ZOOO*U
Based on the modified  elutriate test data,  the effluent suspended  solids concentration cannot exceed
18 mg/L without  exceeding  the  standard.   Similar determinations should also be  made  for  other
contaminants being considered in order to define the  minimum value  for the required  effluent sus-
pended solids concentration.  For this example, 18 mg/L is the minimum value.

An appropriate value should be selected from Table B2 for the resuspension factor.  The  minimum
ponding  depth of  2 ft required by the  site design was selected.   A resuspension  factor of 1.5 was
selected, corresponding to an available area < 100 acres and the selected ponding depth of 2  ft.

The value of 18 mg/L  of SS which must  be achieved at the point  of discharge includes anticipated
resuspension.   The  corresponding value  for total  suspended solids  concentration under  quiescent
settling conditions  is determined as:
or transposed,
                                                                                DRAFT

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                                             B-36
                                      ssa  =  18 mg/L
                                                 .5
                                                        ,,
                                                        12
The disposal area must provide a retention time which will allow the necessary sedimentation.  The
required retention  time to  achieve  12 mg/L under quiescent  settling  conditions  may be determined
from the relationship of suspended solids versus retention time for the laboratory column.  Using the
concentration profile data and the selected depth of ponding at the weir of 2 ft, the relationship for
suspended solids versus field mean retention was developed as was previously shown in Figure BS.
Using Figure B5,  12 mg/L corresponds to a field mean retention tune of 36 h.   To determine the
required disposal site geometry, the theoretical volumetric retention time should be used.  Since  no
other data were available, the hydraulic  efficiency correction  factor was assumed to be 2.2S.  The
theoretical volumetric retention time was calculated as:
                                          •   (HECF)


or transposed,

                              T = Td (HEF) =36  (2.25)  = 81 h
B4.2.5         Determination of Ponded Volume and Surface Area

The required disposal area ponded volume can now be determined using data on anticipated flow rate
and the theoretical volumetric retention time.  Since the dredging equipment  available in the project
area is capable of flow rates up to 15 cfs, the high value  should be assumed.

The ponded volume required is calculated as:
or transposed,
                         V, = JL = 81 Ax 15 cfs  = 100 acre_ft
                                                                               DRAFT

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	B-37	

A ponding depth of 2 ft is the minimum allowed.  This same depth should be maintained over the
entire  ponded  surface area  and  at  the weir.   The disposal site should  therefore  encompass
approximately SO acres of ponded surface area with an average depth of 2 ft if the dredge selected for
the project has  an effective flow rate not greater than 15 cfs.  The surface area of 50 acres required to
meet the water quality standard controls the design  instead of the calculated surface area of 20 acres
required for effective sedimentation.
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	B-38	

B5.0          REFERENCES

Averett, D. E., M.R. Palermo and R. Wade. 1988.  Verification of procedures for design of dredged
       material  containment areas  for  solids retention.   Technical  Report D-88-2, U.S.  Army
       Engineer Waterways Experiment Station, Vicksburg,  MS.

Fulk, R.,  D. Gruber and R. Wullschleger.  1915.  Laboratory study of the release of pesticide and
       PCB materials to the water column during dredging and disposal operations.  Contract Report
       D-75-6, prepared by Envirex, Inc., under contract to the US Army Engineer  Waterways
       Experiment Station, Vicksburg, MS.

Krizek, R.J., J.A.  Fit/Patrick and D.K. Atmatzidis.  1976. Investigation of effluent filtering systems
       for dredged material containment facilities.  Contract  Report D-76-8, prepared by  Dept. of
       Civil Engineering,  Northwestern University, under contract to the U.S.  Army  Engineer
       Waterways Experiment Station, Vicksburg, M.S.

Montgomery, R.L. 1983. Methodology for design of fine-grained dredged material containment areas
       for solids retention.  Technical Report D-78-56, U.S. Army Engineer Waterways Experiment
       Station, Vicksburg,  MS.

Montgomery, R. L., E. L. Thackston and F. Parker.  1983.  Dredged material  sedimentation basin
       design. J. Environ. Engineer, Am. Soc. Civil Engineers.  109.

Palermo, M. R.  1986.  Development of a modified elutriate test for predicting the quality of effluent
       discharged from confined dredged  material  disposal areas. Technical Report D-86-4, U.S.
       Army  Engineer Waterways Experiment Station, Vicksburg, MS.

Palermo, M. R.  1988. Field evaluations of the quality of effluent from  confined dredged material
       disposal  areas.   Technical  Report D-88-1,  U.S. Army  Engineer  Waterways  Experiment
       Station, Vicksburg,  MS.

Palermo,  M. R.  and E. L. Thackston.   1988a.   Test for dredged material effluent  quality.  J.
       Environ. Engineer., Am. Soc. Civil  Engineers  114:  1295-1309.

Palermo,  M. R.  and E. L. Thackston.   1988b.   Verification of predictions of dredged material
       effluent quality. J. Environ. Engineer., Am.  Soc. Civil Engineers  114: 1310-1330.
                                                                              DRAFT

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	B-39	

Palermo, M. R.  and E.L.  Thackston.  1988c.  Refinement of column settling test procedures for
       estimating the quality of effluent from confined dredged material disposal areas.  Technical
       Report D-88-9, U.S. Army Engineer Waterways Experiment Station, Vicksburg, MS.

USAGE.  1987.   Confined Disposal Material.   Engineer Manual  1110-2-5027, Office, Chief of
       Engineers, U. S. Army Corps of Engineers, Washington, D.C.

USACE/EPA. 1992. Evaluating Environmental Effects of Dredged Material Management Alternatives
       - A Technical Framework. EPA842-B-92-008, U.S. Environmental Protection Agency and
       U.S. Army Corps of Engineers, Washington, D.C.
                                                                           DRAFT

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B-40
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     APPENDIXC
EVALUA TION OF MIXING
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                             TABLE OF CONTENTS
                                                                             Page No.

Table of Contents                                                                     i
List of Tables                                                                       iii
List of Figures                                                                       iv

APPENDIX C - EVALUATION OF INITIAL MIXING
Cl.O         INTRODUCTION	  C-l
Cl.l         Background	  C-l
C1.2         Regulatory Considerations  	  C-2
C1.3         Potential Applications of Initial Mixing	  C-2
Cl.3.1       Screen to Determine Need for Additional Water Column Testing  	  C-3
Cl.3.2       Evaluation of Dissolved Contaminant Concentrations by Comparison with
             Water Quality Standards  	  C-3
Cl.3.3       Evaluation of Concentrations of Suspended Plus Dissolved Constituents by
             Comparison with Toxicity Test Results 	  C-3
C1.4         Physical Characteristics of Dredged Material Discharges	  C-4
Cl.4.1       Barge Discharge	  C-4
Cl.4.2       Hopper Dredge Discharge	  C-4
Cl.4.3       Pipeline Dredge Discharge	  C-5
Cl.4.4       Confined Disposal Facility (CDF) Effluent Discharge	  C-5
C1.5         Applicability of Models and Techniques  	  C-6
Cl.5.1       General Considerations  	  C-6
C 1.5.2       Considerations for Tidally Influenced Rivers and Estuaries	  C-7
C 1.5.3       Recommended Models and Techniques  	  C-8

C2.0         SHORT TERM  FATE MODEL  FOR OPEN WATER BARGE  AND
             HOPPER DISCHARGES (STFATE)	  C-10
C2.1         Introduction	  C-10
C2.2         Theoretical Basis	  C-10
C2.2.1       Convective Descent  	  C-ll
C2.2.2       Dynamic Collapse	  C-12
C2.2.3       Transport-diffusion	  C-12
C2.3         Model Capabilities	  C-15
C2.3.1       Disposal Methods  	  C-15
C2.3.2       Ambient Environment  	  C-16
C2.3.3       Time-varying Fall Velocities  	  C-16
C2.3.4       Conservative Constituent Computations	  C-17
C2.4         Model Input	  C-17
C2.5         Model Output	  C-17
C2.6         General Instructions for Running the Model	  C-21
C2.6.1       Target Hardware Environment  	  C-21
C2.6.2       Installation and Starting	  C-22
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                                         11

C2.6J       User Interface	  C-22
C2.7         Steps in Using the Model  	  C-22
C2.8         STFATE Application Examples	  C-29
C2.8.1       Split-hull Barge or Scow Example  	  C-30
C2.8.1.1      Entering STFATE and the Input Data File Selection Menu	  C-30
C2.8.1.1.1           Site Description Data	  C-33
C2.8.1.1.2           Velocity Data	  C-33
C2.8.1.1.3           Input, Execution and Output Keys	  C-3S
C2.8.1.1.4           Material Description Data	  C-35
C2.8.1.1.5           Disposal Operation Data	  C-36
C2.8.1.1.6           Coefficients  	  C-36
C2.8.1.1.7           Saving Input Data Menu	  C-37
C2.8.1.2      Description of Barge Disposal Example Output	  C-37
C2.8.1.2.1           Barge  Disposal  Water   Column   Concentrations   and  Area
                    Distribution	  C-38
C2.8.1.2.2           Barge Disposal Water Column Concentrations  	  C-38
C2.8.1.2.3           Plots  of Concentration Following Barge Disposal	  C-42
C2.8.2       Multiple-bin Hopper Dredge Example	  C-45
C2.8.2.1      Entering STFATE and the Input Data File Selection Menu	  C-45
C2.8.2.1.1           Site Description Data	  C-45
C2.8.2.1.2           Velocity Data for Hopper Disposal Example	  C-48
C2.8.2.1.3           Input, Execution and Output Keys	  C-49
C2.8.2.1.4           Material Description Data	  C-50
C2.8.2.1.5           Operation Data	  C-50
C2.8.2.1.6           Coefficients  	  C-51
C2.8.2.1.7           Saving Input Data Menu	  C-51
C2.8.2.2      Description of Example Hopper Disposal Output	  C-52
C2.8.2.2.1           Hopper  Disposal  Water   Column  Concentrations  and  Area
                    Distribution	  C-53
C2.8.2.2.2           Hopper Water Column Concentrations 	  C-53
C2.8.2.2.3           Hopper Maximum Concentration and Contour Graphs	  C-57

C3.0         CORNELL MIXING ZONE EXPERT SYSTEM  (CORMIX)  	  C-60

C4.0         MACINTYRE  ANALYTICAL METHOD FOR CDF DISCHARGE IN
             RIVERINE CONDITIONS	  C-61
C4.1         Introduction	  C-61
C4.2         Data Requirements  	  C-62
C4 J         Calculation Procedure  	  C-62
C4.4         Example Mixing-Zone Calculation  	  C-66

C5.0         FASTTABS   MODELING   SYSTEM   FOR  EVALUATION   OF
             HYDRODYNAMIC TRANSPORT	  C-72

C6.0         DILUTION VOLUME METHOD FOR CDF EFFLUENT DISCHARGES .  C-73
C6.1         Approach	  C-73
C6.2         Sample Computations	  C-76

C7.0         REFERENCES	  C-78
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                                           Ill
                                 LIST OF TABLES
                                                                               Page No.

Table C-l.    Summary  of Discharge Types, Hydrodynamic Conditions, and Applicable
             Models and Methods for Evaluation of Initial Mixing.                         C-9

Table C-2.    STFATE Model Input Parameters.                                         C-18

Table C-3.    STFATE  Input  Variables  for  Section  404(b)(l)  Regulatory  Analysis  of
             Navigable Waters Using a Scow/Barge Disposal.                            C-31

Table C-4.    STFATE  Input  Variables  for  Section  404(b)(l)  Regulatory  Analysis  of
             Navigable Waters Using a Multiple-bin Hopper Dredge Disposal.              C-46
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IV


| LIST OF FIGURES |

Page No.
Figure C-l.
Figure C-2.
Figure C-3.
Figure C-4.
Figure C-5.
Figure C-6.
Figure C-7.
Figure C-8.
Figure C-9.
Figure C-10.
Figure C-l 1.
Figure C-12.
Figure C-13.
Figure C-14.
Figure C-15.
Figure C-16.
Figure C-17.
Figure C-18.
Figure C-19.
Illustration of Placement Processes.
Velocity Profile Available for Use in PC Model.
Menu Tree for STFATE Model.
STFATE Input Menus.
Schematic of Example Disposal Site for Barge Disposal.
X-direction Velocity Profile for Barge Example.
Selected Output for Barge Disposal.
Peak Lead Concentration in Water Column as a F(Time) for the Barge
Disposal Example.
Lead Concentration Contours for 15 ft Depth at 3600 sec for the Barge
Disposal Example.
Plot of Required Mixing Zone for the Barge Disposal Example.
Schematic of Example Disposal Site for Multiple-bin Hopper Dredge
Disposal.
X-direction Velocity Profile for Hopper Dredge Example.
Schematic of Hopper Dredge with 6 Bins.
Selected Output for Hopper Dredge Disposal.
Peak Fluid Ratio as a F(Time) for the Hopper Dredge Disposal Example.
Fluid Ratio Contours for 20 ft Depth at 3600 sec for the Hopper Dredge
Disposal Example.
Plot of Required Mixing Zone for the Hopper Disposal Example.
Schematic of a Mixing Zone for a Single Effluent Source.
Projected Surface Area and Volume Equations for CDF Effluent Discharge
with Prevailing Current.
C-ll
C-16
C-23
C-24
C-34
C-34
C-39
C-43
C^4
C-44
C-48
C-49
C-51
C-54
C-57
C-58
C-59
C-61
C-74
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                                              C-l

Cl.O          INTRODUCTION

This appendix  presents a variety  of techniques for evaluating the size of mixing zones for dredged
material discharges.  These techniques include analytical approaches and computer models for evalu-
ation of discrete discharges from barges or hoppers, for continuous discharges from pipelines,  and for
effluent discharges from confined disposal facilities (CDFs).  Discussions  of the applicability  and
limitations of the techniques and  stepwise procedures  for performing the required calculations or
applying the models are presented.


Cl.l          Background

Whenever contaminant concentrations in  a  dredged material discharge are above water quality stan-
dards,  there will be some limited initial mixing zone (or zone of dilution) in the vicinity of the dis-
charge point where receiving water quality standards may be exceeded.   The Guidelines recognize
that it  is not possible  to set universal  standards for the acceptable size of mixing zones since receiving
water conditions vary so much from  one  location to  another.   The Guidelines therefore instruct that,
as part of the dredging permit process, the size of any proposed mixing zone should be estimated  and
submitted to the permitting authority.  The permitting authority must then  consider receiving water
conditions at the proposed site and decide if the proposed mixing-zone size is acceptable.

Many state regulatory agencies may specify a limit to mixing-zone dimensions as a condition in grant-
ing the State water quality  certification.  In this  case the mixing zone necessary to meet applicable
standards must be smaller than the specified limits.

The size  of a  mixing zone depends  on a number of factors  including the contaminant  or dredged
material  concentrations in the  discharge,  concentrations in the  receiving water,  the  applicable water
quality standards, discharge density and flow rate, receiving water flow rate and turbulence, and the
geometry  of the discharge vessel, pipeline, or outlet structure and  the receiving water boundaries.
Since the  maximum allowable mixing zone specified by regulatory agencies is usually on the order of
hundreds  of meters, the  evaluation of mixing-zone sizes must necessarily be based on calculation of
near-field dilution and dispersion processes.

There  are a variety of possible estimation  techniques for most real mixing-zone problems, but  any
choice  of a suitable technique involves some tradeoffs. The available techniques may be thought of as
ranging  from  sophisticated computer models,   which  are  sometimes  capable  of very accurate
predictions,  to simple approximations that yield order-of-magnitude estimates. The most sophisticated
models may not run on a microcomputer, and they may require a considerable  amount of measured
                                                                                 DRAFT

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                                               C-2

data and personpower for calibration of the model to a single site.   By contrast, the simplest of
approximations may be made on the basis of several simplifying assumptions and hand calculations.
C1.2          Regulatory Considerations

Any evaluation of potential water column effects has to consider the effects of mixing.  Section 230.3-
(m) of the Guidelines defines the mixing zone as follows:

       The term "mixing  zone" means a limited volume of water serving as a zone of initial
       dilution in the immediate vicinity of the discharge point where receiving water quality
       may  not  meet  quality standards or other requirements  otherwise  applicable to the
       receiving water. The mixing zone should be considered as a place  where wastes and
       water mix and not  as a place where wastes are treated.

Further, Section 230.11(f)  requires that:

       The  mixing  zone  shall  be confined  to the  smallest  practicable zone within each
       specified  disposal site that is consistent with the type  of dispersion determined to be
       appropriate by the  application of these Guidelines. In a few special cases under unique
       environmental  conditions,  where there  is   adequate  justification  to  show  that
       widespread  dispersion  by natural means  will result  in no  significantly  adverse
       environmental effects, the discharged  material may  be intended to be spread naturally
       in a very thin layer over a large area rather than be  contained within the disposal site.
C1.3          Potential Applications of Initial Mixing

        There are three potential applications of initial mixing evaluations:

        a)      screen to determine the need for additional water column testing under Tier II

        b)      evaluate dissolved contaminant  concentrations  by  comparison  with  water quality
               standards after allowance for mixing under Tier II

        c)      evaluate concentrations of suspended plus dissolved constituents  by comparison with
               toxicity test results after  allowance for mixing under Tier III.
                                                                                  DRAFT

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                                              C-3

Cl J.I         Screw to Determine Need for Additional Water Column Testing

The screen determines the necessity for additional water column testing.  This determination is based
on a standardized calculation comparing the bulk  contamination of the dredged material with water
quality standards,  considering the effects of initial  mixing. This "worst case" approach assumes that
all of the contaminants from the dredged material are released to the fluid fraction and subsequently to
the water column.  Mixing evaluations  need only be made for the contaminant requiring the greatest
dilution to meet its  water quality standard.  The  key parameter derived from the evaluation is the
maximum concentration of the  contaminant  in the water column at the boundary of the mixing zone.
This concentration is compared with the applicable water quality standard to determine if additional
water column testing is necessary.  This evaluation cannot be used to predict water column impacts
but only to determine the need for additional water column testing.
Cl.3.2         Evaluation of Dissolved Contaminant Concentrations by Comparison with Water
               Quality Standards

If additional water column testing  is necessary, the potential for water column impacts  may  be
evaluated under  Tier  II by  comparison of  predicted  dissolved  contaminant  concentrations,  as
determined by  an elutriate test,  with the  water quality standards, considering the effects of mixing.
This approach  is used if there are water  quality standards for all contaminants of concern; if these
conditions are not met, the procedure in Section Cl.3.3 is used.   The mixing evaluation need only be
made for the contaminant requiring the greatest dilution to meet its water quality standard.  The key
parameters derived from the model  are the maximum dissolved concentration  of  the contaminant at
the boundary of  the mixing zone.   This  concentration is compared  to the applicable  water quality
standard to determine if the discharge complies with the Guidelines.
Cl.3.3         Evaluation  of  Concentrations  of Suspended  Plus  Dissolved  Constituents  by
               Comparison with Toxicity Test Results

If additional water  column  testing is  necessary,  the potential  for  water column impact  may  be
evaluated under Tier III by comparison of predicted concentrations of the suspended  plus dissolved
constituents of the dredged material with toxicity test results, considering the effects of mixing. For
this case, the dilution of the dredged-material elutriate expressed as a percent of the initial volume of
disposed fluid in a given volume of water column is calculated.  The key parameters derived from the
evaluation are the maximum concentration of dredged-material elutriate in the water  column  at the
                                                                                DRAFT

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                                              C-4
boundary of the mixing zone.  These concentrations are  compared to 0.01 of the LCjo or EC*, as
determined by toxicity tests to determine if the discharge complies with the Guidelines.
C1.4          Physical Characteristics of Dredged Material Discharges

Knowledge  of the physical  characteristics of dredged  material discharges is  necessary for proper
selection of a technique or model for evaluation of initial mixing.  Dredged material can be placed in
open-water sites using direct pipeline discharge, direct mechanical  placement, or release from hopper
dredges or scows.  Discharges of effluent from CDFs can be introduced to the receiving waters in a
variety of ways including direct pipeline outfalls or open channels.  For  purposes of evaluation of
initial mixing, barges or hopper  dredge discharges are discrete discharges, while direct discharge
from a pipeline dredge or CDF effluent should be considered continuous discharges.
Cl.4.1         Barge Discharge

Bucket or clamshell dredges remove the sediment being dredged at nearly its in situ density and place
it on a barge or scow for transportation to the disposal area.  Although several barges may be used so
that the dredging is essentially continuous, disposal occurs as a series of discrete discharges.  Barges
are designed with bottom doors or with a split-hull, and the contents may be emptied within seconds,
essentially as an instantaneous discharge.  Often sediments dredged by clamshell remain in fairly large
consolidated clumps  and reach the bottom in this form.  Whatever its form, the dredged material
descends rapidly through the  water column to the bottom,  and only a small amount of the material
remains  suspended.   Clamshell dredge operations may  also be  used for  direct material  placement
adjacent to the area being dredged. In these instances, the material also falls directly to the bottom as
consolidated clumps.
Cl.4.2        Hopper Dredge Discharge

The characteristics and operation of hopper dredges result in a mixture of water and solids stored in
the hopper for transport to the disposal site.  At the disposal site, hopper doors in the bottom of the
ship's hull are opened, and the  entire hopper contents are emptied in a matter of minutes; the dredge
then returns to the dredging site to reload.  This procedure produces a series  of discrete discharges at
intervals of perhaps one to several hours.  Upon release from the hopper dredge at the disposal site,
the dredged material falls through the water column as a well-defined jet of high-density fluid which
may  contain blocks  of  solid material.  Ambient  water is  entrained during  descent.   After  it hits
                                                                                 DRAFT

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                                              C-5

bottom, most of the dredged material comes to rest.  Some material enters the horizontally spreading
bottom surge formed by the impact and is carried away from the impact point until the turbulence of
the surge is sufficiently reduced to permit its deposition.
C1.4J         Pipeline Dredge Discharge

Pipeline dredges are commonly used for open-water disposal adjacent to channels.  Material from this
dredging operation consists of a slurry with solids concentration ranging from a few grams per liter to
several hundred grams  per liter.  Depending on material characteristics, the slurry may contain clay
balls, gravel, or coarse sand material.  This coarse material quickly settles to the bottom.   The mix-
ture of dredging  site water and finer particles has a higher density than the disposal site water and
therefore can descend to the bottom forming a fluid mud layer.  Continuing the discharge  may cause
the fluid mud layer to  spread. There will be a vertical gradient  of fine suspended solids  forming a
turbidity layer above  the  fluid  mud  layer,  created  by  the discharge momentum  and resulting
turbulence  and entrainment  of disposal site water  into the discharge plume.  The suspended solids
concentration of the fluid mud layer is  typically  10 g/L or greater while the overlying turbidity layer
is defined  as  less than 10 g/L.  Characteristics of the plume are determined  by:   discharge rate,
characteristics of the slurry (both  water and solids), water depth,  currents, meteorological conditions,
salinity of receiving  water, and discharge configuration.
Cl.4.4         Confined Disposal Facility (CDF) Effluent Discharge

Dredged material hydraulically placed in a confined disposal area settles, resulting in a thickened de-
posit of material overlaid by a clarified supernatant.  The supernatant waters are discharged from the
site  as  effluent  during active dredging  operations.   The  effluent  may  contain  both dissolved
contaminants and  suspended colloidal particles with associated (adsorbed or held by ion exchange)
contaminants.  Supernatant waters from confined disposal sites  are discharged after a retention time of
up to several days.  Furthermore, actual withdrawal of the supernatant is governed by the hydraulic
characteristics of the ponded area and the discharge weir. The  effluent  suspended solids concentration
is typically less than 100 mg/L for sediments dredged  from estuarine environments  and less than a
few grams per liter for sediments dredged from freshwater environments.
                                                                                 DRAFT

-------
                                              C-6

C1.5          Applicability of Models and Techniques

Cl.5.1         General Considerations

Equations can be derived from a simplistic approach to the problem of estimating mixing-zone size
that  make  it possible to  use a combination of empirical  and  analytical  solutions.   However, the
simplifications  that make the calculations easily manageable are somewhat restrictive, and a  more
advanced set of similar empirical and analytical solutions could be used to estimate mixing-zone sizes
under more  complex conditions.   The more  advanced  analytical  solutions  involve many  more
computations, and  for this reason they  are more easily  dealt  with  by use of  a  computer.   The
simplicity and limited data requirements of analytical solutions make them  an attractive tool.  How-
ever, analytical solutions cannot be used for receiving water where there are complex hydrodynamic
conditions,  nor can  they be  applied  under dynamic (unsteady) flow  conditions.   Where  these
conditions  exist, a numerical model  must be used, and numerical  dispersion  models  are not
susceptible to hand calculation.  In addition to requiring  a computer solution  technique, numerical
models generally require a much more detailed set of input data, and the collection  of such data can
be expensive.

No models have been identified that are suitable for a broad range of mixing  zone conditions, and
there are no  readily available models suitable for modeling the first few hundred metres downstream
from the  discharge point.   This is because the overwhelming  majority  of computer  models are
concerned  with far-field  solutions  where  concentrations  can be  adequately described  by  a  two-
dimensional or a one-dimensional model and the initial characteristics of the discharge are relatively
unimportant. These models are generally inadequate  in the immediate vicinity of a discharge, where
a three-dimensional description of concentrations is often necessary and where the initial  characteris-
tics of the discharge can be highly significant.  Within the  first few hundred metres of the discharge,
there are  several different processes that may be  significant,  so a general model  must be  able to
estimate each of the  processes (for  example, momentum,  buoyancy, dispersion) and  to  identify the
zones within which the processes are dominant.  A general mixing-zone model must therefore be a
series of submodels,  each of which can handle a zone that is  dominated  by  one of the principal
mixing processes. The sub-models must be capable of determining the limits of their applicable zones
and passing concentration values at these limits on  to other submodels so that the entire mixing zone
may be estimated.  The following tabulation presents a summary of the steady-state physical processes
that  might be  suitable for inclusion as submodels in a general mixing-zone model.  Sources that
presently seem to present the most  promising  empirical and analytical solutions to these sub-model
processes are also presented in the tabulation.
                                                                                DRAFT

-------
                                              C-7
  Physical Process to be Handled by a Submodel	Source

  Momentum and/or buoyancy-dominated jets       Zeller et al. (1971)
                                                 Motz and Benedict (1972)
                                                 Buhler and Hauenstein (1981)
                                                 Jirka et al.  (1981)
                                                 Wright (1984)
 	Doneckar and Jirka (1990)

  3-dimensional dispersion in receiving water       Prakash (1977)
                                                 Fischer et al. (1979)
                                                 Johnson et  al. (1994)
 	King (1992)	

  2-dimensional vertically averaged dispersion       Stefan and  Gulliver (1978)
                                                 Paily and Sayre (1978)
                                                 Gowda (1984a, b)
                                                 Thomas and McAnally (1990)
Cl.5.2        Considerations for Tidally Influenced Rivers and Estuaries


The assumptions necessary for evaluation of mixing are more difficult to satisfy in estuaries and the
tidally influenced portions of rivers. The assumption that velocities in the water body near the mixing
zone can be represented by a single mean velocity parallel to the bank is usually a reasonable one in
the non-tidally influenced  portion  of  a river.  However,  it is not always acceptable in  estuaries.
Typically the downstream  section  of an estuary exhibits horizontal circulation patterns, so that the
horizontal water velocity and direction vary with distance parallel to the bank, distance perpendicular
to the bank,  and time.  Under these conditions, water near the mixing zone may not always travel
parallel to the bank.   Therefore, simple mixing-zone equations may  not be applicable to  the wide,
open low-velocity sections of estuaries.


Also, mixing-zone equations  are not theoretically applicable as the mean velocity tends to zero.  This
is because the equations are dependent upon the process of advection, which does  not  exist in the
absence of a flow velocity, and also because the primary source of dispersion is assumed  to be the
turbulence caused by the horizontal movement of water.   However,  in a real water body, as  the
velocity tends to zero, the primary sources of turbulence and dispersion are die wind and waves.


The rate of change of water velocity due to tidal effects can also cause problems.   The time taken for
material to travel the length of the mixing zone should be an order of magnitude smaller than the time
taken for a 10-percent change in the mean water velocity. It may be possible to satisfy this  condition
                                                                                DRAFT

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                                              C-8

in a river, but it will probably not be possible to do so in most estuaries during a significant portion
of the tidal cycle.

Another potential difficulty in estuaries is the phenomenon of stratification.  Estuaries with low water
velocities sometimes have a layer of relatively fresh water near the surface with a much more saline
denser layer of water near the bottom and with quite a distinct interface between the two layers.  The
abrupt change of density at the interface tends to inhibit  vertical mixing through the entire depth of
the water column.
Cl.5.3         Recommended Models and Techniques

Several  models and approaches for evaluation of initial mixing are provided in this appendix.  Table
C-l provides a summary of the characteristics of the various types of dredged material discharges,
hydrodynamic environments, and the models recommended for use in evaluation of initial mixing for
those conditions.  Descriptions of each of the models  and details on applying the models are provided
in the following sections of this appendix.
                                                                                DRAFT

-------
                                             C-9
Table C-l.    Summary of Discharge Types, Hydrodynamic Conditions, and Applicable Models and
              Methods for Evaluation of Initial Mixing.
Type of
Discharge
BARGE
HOPPER
PIPELINE

CDF
EFFLUENT


Characteristics
of Discharge
Discrete
Semi-Discrete
Continuous

Continuous


Near-Field
Effects
Strong
Moderate
Moderate

Weak


Applicable
Model or
Technique
STFATE
STFATE
CORMIX1
TABS2
Maclntyre
TABS2
Dilution
Volume
Method
Model
Hydrodynamics
Steady
Non-uniform
Steady
Non-uniform
Steady
Uniform
Unsteady
Non-uniform
Steady
Uniform
Unsteady
Non-uniform
Steady
Uniform
Section
C2.0
C2.0
C3.0
C5.0
C4.0
C5.0
C6.0
1 CORMIX has not been developed and verified for national application. However,  the fundamental
processes contained in CORMIX are applicable for continuous pipeline discharges and this model is
currently under investigation for future use.

2 TABS has not been developed  and verified for national  application  for the indicated discharges.
However, the  fundamental far-field  processes contained  in TABS are applicable for the indicated
discharges and this model can be adapted for use on a regional basis. Note that the TABS model
computes far-field effects only. Some independent near-field  analysis is usually required.
                                                                              DRAFT

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                                           C-10

C2.0          SHORT  TERM FATE MODEL FOR OPEN WATER  BARGE AND HOPPER
              DISCHARGES (STFATE)

C2.1          Introduction

The model described in this section is the STFATE (Short-Term FATE of dredged material disposal
in open  water)  model  (Johnson  et  al., 1994)  developed  from  the  DIFID  (Disposal  From  an
Instantaneous Discharge) model originally prepared by Koh and Chang  (1973).   This model is used
for discrete discharges from barges and hoppers.  STFATE is a module of the Automated Dredging
and Disposal Alternatives Management System (ADDAMS) (Schroeder  and Palermo, 1990) and can
be run  on DOS-based  personal  computers  (PC) having 80386  or higher  processors with  math
coprocessors.  ADDAMS is an interactive computer-based design and analysis system in the field of
dredged-material management. The general goal of ADDAMS is to provide state-of-the-art computer-
based tools that will increase the accuracy,  reliability,  and cost effectiveness of dredged-material
management activities in a timely manner.

An executable version of the STFATE model for use on IBM-compatible microcomputers is found on
the floppy disk in the  pocket inside the back  cover of  this manual.  The  disk contains  a model
appropriate for instantaneous discharges from  barges or scows and sequential discharges from hopper
dredges.
C2.2          Theoretical Basis

The behavior of the material during disposal is assumed to be separated into three phases:  convective
descent, during which the disposal cloud falls under the influence of gravity and its initial momentum
is imparted by gravity; dynamic collapse,  occurring when  the descending cloud either impacts the
bottom or arrives at  a level of neutral buoyancy where descent is retarded and horizontal spreading
dominates; and passive transport-dispersion, commencing when the material transport and spreading
are determined more by ambient  currents and turbulence than by  the  dynamics  of the disposal
operation. Figure C-l illustrates these phases.
                                                                             DRAFT

-------
                                            C-ll
            vCg^ J9- Ji.TW-'.S
                CQNVECTIVE
                 DESCENT
                     DYNAMIC COLLAPSE
                         ON  BOTTOM
LONG-TEEM
  PASSIVE
                          BOTTOM
                        ENCOUNTER
                                    DIFFUSIVE SPREADING  DIFFIISIOM
                                       GREATER THAN
                                     DYNAMIC SPREADING
Figure C-l.   Illustration of Placement Processes.
C2.2.1
Convective Descent
In STFATE,  multiple convecting clouds that maintain a hemispherical shape during  convective
descent are assumed to be released. By representing the disposal as a sequence of convecting clouds
released at a constant time interval during the total time required for the material to leave the disposal
vessel,  real disposal operations can be more accurately simulated.  For example, a moving hopper
dredge disposal can be modeled by assuming that the material in each bin convects downward as one
cloud.  In addition, through the  use of multiple convecting clouds with varying characteristics the
consolidation that often occurs in scows or barges  can be accounted for more accurately.  Since the
solids concentration in discharged  dredged material is usually low, each cloud is expected to behave
as a dense liquid; thus, a basic  assumption  is that a buoyant thermal analysis is appropriate.  The
equations  governing the  motion are those for conservation of mass,  momentum, buoyancy,  solid
particles,  and  vorticity.  These equations  are  straightforward statements  of conservation  principles;
details are presented in Koh and Chang (1973)  and  Brandsma and Divoky (1976). It should be noted
that the entrainment coefficient associated with the entrainment of ambient fluid into a  descending
hemispherical cloud is assumed to vary smoothly between its value for a vortex ring and the value for
                                                                              DRAFT

-------
                                              C-12

turbulent thermals.  Model results are relatively sensitive to the entrainment coefficient, which in turn
is dependent upon the material being disposed (the higher the moisture content, the larger the value of
the entrainment coefficient).   Laboratory  studies by Bowers  and  Goldenblatt  (1978) resulted  hi
analytical expressions  for  the  entrainment, drag, and added mass coefficients as  functions of the
moisture content.  These have been incorporated into STFATE.  As these clouds move downward,
material  and fluid with dissolved  contaminants may be stripped away.  Stripped  material is handled
through the concept of Gaussian clouds discussed below.  The amount of material stripped away and
stored in the Gaussian clouds is computed as a coefficient tunes the downward velocity of the cloud
times the cloud surface  area.  The value of the "stripping"  coefficient is selected so that approxi-
mately 2-5 percent of the total  volume of fine material is stripped away at disposal sites of 100 ft or
less.  Based upon field data collected by Bokuniewicz et al. (1978), this will result  in the amount of
stripped material being on the conservative side.
C2.2.2         Dynamic Collapse

Whether by disposal from a split-hull barge or scow or discharge from a multi-bin hopper dredge, the
disposed material  cloud grows  during convective  descent  as  a  result of entrainment.   Eventually,
either the material reaches the bottom, or the density difference between the discharged material and
the ambient water column becomes  small enough for  a position  of neutral buoyancy to be assumed.
In either case, the vertical motion is  arrested and a dynamic horizontal spreading occurs.

The basic  shape assumed for each  collapsing  cloud is an oblate spheroid  if collapse occurs in the
water column,  whereas a general ellipsoid is assumed for  collapse on a sloping bottom.  With the
exception of vorticity,  which is assumed to have been dissipated by  the stratified ambient water
column,  the same conservation  equations used in  convective descent  but now  written for either an
oblate spheroid or an ellipsoid  are applicable.  For the case of collapse on the bottom, a factional
force between the  bottom and the collapsing cloud is included which accounts for energy dissipation
as a result of the  spreading.  Other than the changes noted above, the same equations presented in
Brandsma and Divoky (1976) apply.
C2.2.3        Transport-diffusion

When the  rate of spreading in the dynamic collapse phase becomes less than an  estimated rate of
spreading due to turbulent diffusion in both the horizontal and vertical directions, the collapse phase is
terminated. Laboratory experiments by Johnson et al. (1994) as well as field data collected by Kraus
(1991) imply that fine material is lost to the water column at the top of the collapsing cloud.  As these
                                                                                 DRAFT

-------
                                              C-13

particles leave the main body of material, they are also stored in small  clouds that are characterized
by a Gaussian distribution, i.e.,
                        m
                                exp
- x0)2 .  (y - y,)3
                                                                                            (1)
                   (2tr)Moxayo,

where
           m  = volume of solids in the cloud, ft3
      ffx»ffy»ff* = standard deviations, ft
        x,y,z  = spatial coordinates, ft
      xo>y<»zo  = coordinates of cloud centroid, ft

At the end of each time-step, each cloud is advected horizontally by the input velocity field.  The new
position of the  cloud centroid is determined by
                                       *„_ = X0- + U  • At


                                       Zo_ - ^ + W  ' At                                    (2)

where
     u,w =  input ambient velocities, fps
      At  =  long-term time-step, sec

In addition to the advection or transport of the cloud, the cloud grows both horizontally and vertically
as a result of  turbulent diffusion.  The horizontal diffusion  is based  upon the commonly assumed
four-thirds power law.   Therefore,  the diffusion coefficient,  Kx^  ,  (up  to a maximum  value of
100 ft2/s) is given as

                                                                                            (3)
where  AL  is an input dissipation parameter and  L   is set equal to four standard deviations.  As
illustrated in Figure 2.4 of Brandsma and Divoky (1976),  a value of 100 ft2/sec for the horizontal
diffusion coefficient corresponds to a length  scale of 10M04  feet.  With the computational  grid  cell
typically being on the order of 100-500 ft, a length scale greater than  1,000 ft  would normally be
associated with mean flow rather than turbulence.  Thus, restricting  the diffusion coefficient to  less
than 100 rYVsec is reasonable.

Horizontal growth is achieved by employing the Fickian expression
                                                                                 DRAFT

-------
                                             C-14


                                        °x,x  = (2K^t)1/2                                   (4)

where
     a^ = a standard deviation
       t  = time since formation of the cloud
From Equation 4,
                                      do.
                                                                                          ( 5 )
and thus,
                                                   JV
                                                                                          (6)
where
     a,^.  = a^ at the current time step, At
         = a^ at the previous time step, At
In a similar manner, the vertical growth is written as
where  Ky   is a function of the stratification (including  the effect  of the  sediment) of the water
column.   The maximum value of  Ky  is input  as  a model coefficient and occurs when the water
density  is uniform.   It should be  noted  that since  computations are made for  each solid fraction
independently from the remaining material, the effect of the total volume of suspended material on
reducing vertical diffusion is  not modeled.  This can sometimes lead to confusing  results; e.g.,  a
small amount of sand may become diffused over the  entire water column while a much larger amount
of silt might have its vertical  diffusion suppressed due to the larger concentration.  Modifications to
correct this problem are under investigation.

If long-term output is desired at the end of a particular time-step,  the concentration of each solid type
is given at each grid point by summing the contributions from individual clouds to yield
                                                                                DRAFT

-------
                                              C-15
N
Jp
Ll
mi
1 £>vp
(W*,, exp
1
(x ~ X
-------
                                             C-16
C2.3.2
Ambient Environment
As  illustrated in Figure C-2,  time-invariant  velocity profiles that  allow  for flow  reversal can be
prescribed.  These profiles are  applied at each grid point.  Another option is to specify a time-
invariant, spatially varying depth-averaged velocity.   The ambient density profile at the deepest point
on the grid must also be prescribed.
        DU2
                                                                              DW2
Figure C-2.    Velocity Profile Available for Use in PC Model.
C2.3.3
Time-varying Fall Velocities
If a solid fraction is specified as being cohesive, the settling velocity is computed as a function of the
suspended sediment concentration of that solid type.  The algorithm used is
              v.
            0.000034  if  C < 25 mg/L
            0.0000225 + 1.6 x 10'7 C4/3  if  25 < C < 3000 mg/L
            0.0069  if C > 3000 mg/L
(9)
where
     V, = settling velocity, fps
      C = suspended sediment concentration, mg/L
This approach is taken from Ariathurai et al. (1977).
                                                                               DRAFT

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                                              C-17

C2.3.4         Conservative Constituent Computations

STFATE allows for the dredged material to contain a conservative constituent with perhaps a nonzero
background  concentration  of  that  constituent.    Computing  the  resultant time-history  of that
concentration provides information on the dilution that  can be expected over a period of time  at the
disposal site and enables the computation of mixing zones in water column evaluations.


C2.4          Model Input

Input data for the model are grouped into the following general areas:  (1) description of the disposal
site, (2) description of site velocities, (3) controls for  input, execution, and output, (4) description of
the dredged materials, (5) description of the disposal operation, and (6) model coefficients.

Ambient  conditions  include  current  velocity,  density  stratification, and  water  depths  over  a
computational grid. The dredged material is assumed to  consist of a number of solid fractions, a fluid
component, and conservative dissolved  contaminants.   Each  solid fraction has to  have a volumetric
concentration, a specific gravity, a settling velocity, a void ratio for bottom deposition,  critical  shear
stress, and information  on whether or  not the fraction is cohesive  and/or  strippable.   For initial-
mixing calculations, information on initial concentration, background  concentration, and  water quality
standards  for the constituent to be modeled  have to  be specified.   The description of the disposal
operation  includes  the  position of the disposal barge or hopper dredge on the grid;  the  barge  or
hopper dredge velocity, dimensions, and draft; the  volume  of dredged material  to be discharged.
Coefficients are required for the model  to accurately  specify entrainment,  settling, drag, dissipation,
apparent mass, and density  gradient differences. These  coefficients have default values that should  be
used unless other site-specific information is available. Table  C-2  lists the necessary input parameters
with  their corresponding  units.   Table C-2  also lists the  input parameters for determining  the
contaminant  of concern to be  modeled  based on dilution needs.   More detailed descriptions and
guidance for  selection of values for many of the parameters is provided directly online in the system.
C2.5          Model Output

The output  starts  by echoing the input data and then optionally presenting the time history of the
descent and collapse phases.  In descent history the location of the cloud centroid, the velocity of the
cloud centroid, the radius of the hemispherical cloud, the density difference between the cloud and the
ambient water, the conservative  constituent concentration and the total volume and concentration of
each solid fraction are provided as functions of time since release of the material.
                                                                                 DRAFT

-------
                                                 C-18
Table C-2.     STFATE Model Input Parameters.
Parameter
Disposal*
Operation
Types
                                                                         Units
Options**
rVmtflminflnf Selection Data

Solids concentration of dredged material
Contaminant concentration in the
  bulk sediment
Contaminant concentration in the
  elutriate
Contaminant background concentration
  at disposal site
Contaminant water quality standards

Site Description

Number of grid points (left to right)
Number of grid points (top to bottom)
Spacing between grid points
  (left to right)
Spacing between grid points
  (top to bottom)
Constant water depth
Roughness height at bottom of
   disposal site
Slope of bottom in x-direction
Slope of bottom in z-direction
Number of points in density profile
Depth of density profile point
Density at profile point
Salinity of water at disposal site
Temperature of water at disposal site
Grid points depths

Velocity Data

Type of velocity profile
Water Depth for Averaged Velocity
Vertically averaged x-direction
   velocity
Vertically averaged z-direction
   velocity
Water depths for 2-point profile
Velocities for 2-point profile in
   x-direction
Velocities for 2-point profile in
   z-direction
Velocities for entire grid in
   x-direction
Velocities for entire grid in
   z-direction

Input. Execution and Output Keys

Processes  to simulate
Duration of simulation
Long-term time step for diffusion
Convective descent output option
H,B
H,B

H,B

H, B
H,B

H, B
H,B
H,B
H, B
H, B
H,B
H, B
H,B
H, B
                                                        H, B
                                                        H, B

                                                        H, B

                                                        H, B
                                                        H, B

                                                        H, B

                                                        H,B

                                                        H,B

                                                        H,B
                                                        H,B
                                                        H,B
                                                        H, B
                                                        H,B
                                                                         g/L

                                                                         Mg/Kg

                                                                         /tg/L
                                                                         ft

                                                                         ft
                                                                         ft

                                                                         ft
                                                                         degrees
                                                                         degrees
                                                                         ft
                                                                         g/cc
                                                                           , •
                                                                           lsius
                                                                         ft
Optional
Optional
V
                 ft

                 ft/sec

                 ft/sec
                 ft

                 ft/sec

                 ft/sec

                 ft/sec

                 ft/sec
                 sec
                 sec
                                                                                       DRAFT

-------
C-19
TaKl* CJJ STFATR Mrvfol Tnniit PQ ram Ate re frrmt
Parameter
Input. Execution "W^ Output Kevs (continued)
Collapse phase output option
Number of print times for long-term
diffusions
Location of upper left corner of
mixing zone on grid
Location of lower right comer of
mixing zone on grid
Water quality standards at border of
mixing zone for contaminant of
concern
Contaminant of Concern
Contaminant concentration in sediment
Background concentration at disposal
site
Location of upper left corner of
zone of initial dilution (ZID)
on grid
Location of lower right corner of
zone of initial dilution (ZID)
on grid
Water quality standards at border of
ZID for contaminant of concern
Number of depths in water column
for which output is desired
Depths for transport - diffusion output
Predicted initial concentration in
fluid fraction
Dilution required to meet toxicity
standards
Dilution required to meet toxicity
standards at border of ZID
Material Description Data
Total volume of dredged material in
the Hopper dredge
Number of distinct solid fractions
Solid-fraction descriptions
Solid-fraction specific gravity
Solid-fraction volumetric
concentration
Solid-fraction fall velocity
Solid-fraction deposited void ratio
Solid-fraction critical shear stress
Cohesive? (yes or no)
Stripped during descent? (yes or no)
Moisture content of dredged material
as multiple of liquid limit
Water density at dredging site
Salinity of water at dredging site
Temperature of water at dredging site
Desired number of layers
Volume of each layer
Velocity of vessel in x -direction
during dumping of each layer
Velocity of vessel in z-direction
during dumping of each layer
in,,«n
Disposal*
Operation
Types

H, B
H,B

H, B

H, B


H,B
H, B
H, B

H, B


H, B


H,B

H, B

H,B
H, B

H, B

H,B

H, B


H
H,B
H,B
H,B
H,B
H, B
H,B
H, B
H, B
H, B
H, B
H, B
H, B
H,B
B
B

B

B

Units




ft

ft


mg/L

mg/Kg

mg/L


ft


ft

mg/L


ft

mg/L

percent

percent


yd3


yd3/yd3
ft/sec

Ibs/sq ft



g/cc
ppt
Celsius

yd3

ft/sec

ft/sec

Option**










































Cohesive

Optional
Optional






                         DRAFT

-------
                                                  C-20
Table C-2.    STFATE Model Input Parameters (continued)
Parameter
Disposal1"
Operation
Types
Units
Option"
Disposal Operation Data

Location of disposal point from
   top of grid
Location of disposal point from
   left edge of grid
Length of disposal vessel bin
Width of disposal vessel bin
Distance between bins
Pre-disposal draft of Hopper
Post-disposal draft of Hopper
Time required to empty all Hopper bins
Number of Hopper bins opening
   simultaneously
Number of discrete openings of sets
   of Hopper bins
Vessel velocity in x-direction during
   each opening of a set of Hopper bins
Vessel velocity in z-direction during
   each opening of a set of Hopper bins
Bottom depression length in x-direction
Bottom depression length in z-direction
Bottom depression average depth
Pre-disposal draft of disposal vessel
Post-disposal draft of disposal vessel
Time needed to empty disposal vessel

Coefficients

Settling coefficient
Apparent mass coefficient
Drag coefficient
Form drag for collapsing cloud
Skin friction for collapsing cloud
Drag for an ellipsoidal wedge
Drag for a plate
Friction between cloud and bottom
4/3 Law horizontal diffusion
   dissipation factor
Unstratified water vertical
   diffusion coefficient
Cloud/ambient density gradient ratio
Turbulent thermal entrainment
Entrainment in collapse
Stripping factor
H,B

H,B
H,B
H, B
H
H
H
H

H

H

H

H
H, B
H, B
H, B
  B
  B
  B
H, B
H, B
H, B
H, B
H,B
H,B
H,B
H, B

H, B

H, B
H, B
H, B
H,B
H, B
ft

ft
ft
ft
ft
ft
ft
sec
ft/sec

ft/sec
ft
ft
ft
ft
ft
sec
Optional
Optional
Optional
 *      The use of a parameter for disposal operations by a multiple bin hopper dredge is indicated in the table
        by an H while a parameter used for disposal from a split-hull barge or scow is indicated by a B.

 **     The use of a parameter for the constant depth option or variable depth option is indicated in the table
        by a C or V, respectively. Other optional uses for parameters are so indicated.
                                                                                        DRAFT

-------
                                             C-21

At  the  conclusion  of the collapse phase,  time-dependent  information  concerning the size of the
collapsing cloud, its density, and its centroid location and velocity as well as contaminant and solids
concentrations can be requested.  The model performs the numerical integrations of the governing
conservation equations in the descent and collapse phases  with a minimum  of user input.   Various
control parameters that give the user insight into the behavior of these computations are printed before
the output discussed above is provided.

At  various  times,   as  requested through input  data,  output concerning  suspended   sediment
concentrations can  be obtained from  the transport-diffusion computations.    With Gaussian cloud
transport and diffusion, only concentrations at the water depths requested are  provided at each grid
point.

For evaluations of initial mixing, results for water column concentrations  can be computed in terms of
milligrams  per  liter of  dissolved  constituent for Tier  II evaluations  or  in  percent  of  initial
concentration of suspended plus dissolved constituents in the dredged material for Tier III evaluations.
The maximum  concentration within the grid and  the  maximum concentration  at  or outside the
boundary  of the disposal site are tabulated  for  specified time intervals.  Graphics showing the
maximum concentrations inside the disposal-site boundary and anywhere on the grid as a function of
time can  also be generated.  Similarly,  contour  plots of concentration can be generated at the
requested water depths and at the selected print times.
C2.6          General Instructions for Running the Model

C2.6.1         Target Hardware Environment

The system is designed for the 80386 based processor class of personal computers using DOS.  This
does not constitute official endorsement or approval  of these commercial products.   In general, the
system requires a math coprocessor,  640  KB of RAM and  a hard  disk.  The STFATE executable
model requires about 565 KB of free  RAM to run; therefore, it may be necessary to unload network
and TSR software prior to execution.   The model  is written primarily  in Fortran 77 but some of the
higher-level operations and file-management operations are written in BASIC and some of the screen
control operations in the Fortran 77 programs are performed using  an Assembly  language utility
program.
                                                                                DRAFT

-------
                                             C-22

C2.6.2         Installation and Starting

All files contained  on the diskettes in the folder in the back of this manual should be saved in a
directory on the hard disk  dedicated  for the STFATE module, e.g. C:\STFATE.   The files are
archived on the diskettes  and have to be dearchived prior to running the model.  To dearchive the
files, copy the files from each diskette onto the hard drive, call up the README file, and follow the
instructions.


C2.6J         User Interface

The STFATE module of ADDAMS employs a menu-driven environment with a full-screen data entry
method.  In general, single keystrokes (usually the  Fl  through F10 function keys, the number keys,
Esc key or  the arrow keys  and the Enter  key)  are required to  select menu options in the system.
Menus are displayed  on the  screen.  Cursor keys  are used to select from among highlighted input
fields (displayed in  reverse video) much like a spreadsheet program.  To enter alphanumeric data, the
user moves the cursor to the  cell of interest, using the up and down arrows to move,  respectively, up
and down,  the  Tab and Shift-Tab keys to move, respectively, right and left.  The Enter key  is also
used to  move forward through the cells.  The left and  right arrow keys are used to move the  cursor
within a selected cell to edit the cell's contents.  The Backspace key is used to clear a single character
in a cell.  The  spacebar will insert a space in alphanumeric cells.  The PgDn key advances the  cursor
to the next data entry screen and the PgUp key returns  control to the previous data entry screen. The
Esc key returns control  to exit to the previous menu without loss of data.  The Home key permits the
user to  exit from the current data entry screen to the Main Menu for the  application without loss of
data.

Results  from computations are generally displayed  in tabular format on the  screen and/or written to
print files or devices.


C2.7           Steps in Using the Model

The menu-driven environment for applying the model is illustrated in Figures C-3 and C-4.

The general steps and menus  used in applying the model for a disposal operation are as follows:
                                                                                DRAFT

-------
                                                                 C-23
                 ADOAMS AppKoation S.l.o(«m M.nu

       1. SETTLE  . CMfhlW Dlspesal Faeilillss (COFs) Design.
       2. DYECOM - Hydraulic Heteation an* Efttoleacy el COft.
       S. PC DDF  • Consolidation and Daiioadmit of Dialed Fill.
       4. STFATE  • Short-Twin Fate of Disposal In Open Water.
       E. D2M2    - DMdgsd Mslerlal Disposal MaaafemeM.
       I. EFQUAL • Modified Elutriate Test Aaalyslt.
       7. WCT     - Wetlands ivahuiio* Tsolink)ue.
       t. U«t data fll« *««•> lor «H eppllotfloiis.
       9. Pvrfoim hardware eenfiouratlon foi graphic*.
       E»o End ourr.nl ADOAMS Milieu.
         STFATE Ey.lualion S»l«rti
-------
                                                          C-24
                                       STFATE Evaluation Selection Menu

                          F1  - General Open Water Disposal Analyst*.
                          F2  - Section 103 Regulatory Analysis for Ocean Waters.
                          F3  - Section 404(b)(1) Reg. Analysis (or Navigable Waters.
                          F4  - Determine Contaminant of Concern Baaed on Dilution Needs.
                         Esc - Return to STFATE Activity Selection Menu.
                              F1
                              F2
                              F3
         STFATE Input Data File Selection Menu

     F1 . Enter name of Input dale III* to be bull! or edited.
     F2 • Enter DOC p«th to data III* storage location (Optional)
     F3 - Display directory of Input data files.
     F4 • Build or edit input data file.
     Eic • Return to STFATE Activity  Selection Menu
                      F4
                    F4
     STFATE Contaminant Data File Selection Menu

 F1  • Enter name of data lite 10 b» built or edited.
 F2  - Enter DOS path to daw flic ttorafle location. (Optional)
 F3  - Display directory ol contaminant data files.
 F4  - Build or edit contaminant data III*.
Eso - Return to STFATE Evaluation Selection Menu.
             Disposal Operation Selection Menu

      F1  - Disposal from a Multiple Bin Hopper Dredge.
      F2 - Disposal from i Split-Hull Barge or Scow.
     Esc - Stop Input and return to STFATE Activity Menu.
                                                                                                  F4
   Menu for Selection of a Contaminant for Modeling

F1  - Build or edit bulk sediment quality data.
F2  - Build or edit elutriate water quality data.
F3  - Compute bulk sediment dilution*.
F4  - Compute elutriate dilution*.
F5  - Save bulk sediment and elutriate water quality data.
Esc - Quit.
              STFATE Input Selection Menu

       F1 - Site Description.
       F2 • Velocity Data.
       F3 • Input, Execution and Output Keys.
       F4 - Material Description Data.
       FS • Disposal Operation Data.
       FS - Coefficients (Default Values).
       F7 - Saving Input Data Menu.
      Eso - Stop input and return to STFATE Activity Menu.
                                                                                                 F5
    STFATE Contaminant Data File Saving Menu

 F1  - Enter name ol file to be saved.
 F2  - Enter DOS path for data tile. (Optional)
 F3  - Display directory of Input data files.
 F4  - Save data In (or to) the active data file.
 Eac - Relurn to STFATE  Evaluation Selection Menu.
                      F7
             STFATE Input File Saving Menu

         Ft - Enter name of file to be saved.
         F2 - Enter DOS path for data file  (Optional)
         F3 - Dliplay directory of Input data tiles.
         F4 - Save data in (or to) the active file.
        Eso - Return to STFATE input Selection Menu
Figure C-4.     STFATE Input Menus.
                                                                                                       DRAFT

-------
                                           C-25

a.      Starting

       Change the directory to make directory containing the STFATE module the default directory.
       Start the program by  entering ADDAMS or STFATE at the DOS prompt.  If started by
       entering  ADDAMS, the program will display first the ADDAMS logo  and  then  an
       Application Selection Menu.  An  application in the ADDAMS software consists of one or
       more standalone computer programs or numerical models for performing a specific analysis.
       The only  ADDAMS application module provided on diskette  with  this  manual is named
       STFATE.   STFATE consists of  programs  for  evaluating  open-water  disposal  of dredged
       material.   Select the STFATE  application module from the Application Selection Menu. The
       module will display  some logos and then a reference screen with points of contact.  After the
       user strikes any key, the module displays the STFATE  Activity Selection Menu.  If started by
       entering STFATE,  the  module starts with  STFATE logos and  the reference  screen  and
       proceeds in the same manner as if the module was started by entering ADDAMS.

b.      Activity Selection Menu

       The activity selection menu may be considered the main menu for the STFATE  application.
       The first option is used  to build or edit an input data file.  The second option executes the
       simulation. The third option  is used to print or  view output.  The fourth option generates
       graphics.   The fifth option is used  to configure the graphics software  for the hardware
       present.

c.      Evaluation Selection Menu

       Selecting Fl from the STFATE Activity Selection Menu  brings up the  STFATE Evaluation
       Selection Menu.  There are  four options available.  The two options of interest here are F3 -
       Section 404(b)(l)  Reg.  Analysis for  Navigable Waters  (see  step  g)  and F4 - Determine
       Contaminant of Concern Based on Dilution Needs (see step d).  This option  is used only for
       Tier II evaluations.

d.      Contaminant Data File Selection Menu

       Selecting F4 from the Evaluation  Selection Menu brings  up this menu  which has the same
       structure as the Input Data File Selection Menu (see step g).  Use one of the options to select
       an active file.
                                                                            DRAFT

-------
                                              C-26

e.     Menu for Selection of a Contaminant for Modeling

       Selecting F4 from the Contaminant Data File Selection Menu brings up this  menu.  A data
       analysis routine controlled by this menu is used to select a specific contaminant for modeling.
       Such a selection is necessary under the Tier II analysis both for evaluation of the need for
       additional testing  and for water quality comparisons with standards.  Execution of the  open-
       water disposal model for these Tier II analyses allows use of only one contaminant; this
       option is used to select that contaminant.

       Bulk sediment and background  contaminant concentrations and  water quality standards are
       required to compute the required dilutions for the evaluation of the need for additional testing.
       The contaminant requiring the largest dilution should be subsequently modeled.

       Elutriate and background concentrations and water quality  standards are required to compute
       the required dilutions for the dissolved contaminants.  The contaminant requiring the largest
       dilution should be subsequently modeled in the Tier II water quality analysis.

f.     Contaminant Data File Saving Menu

       This menu has  the   same structure as the Input  File Saving Menu (see  step j).   The
       contaminant data files are saved with an extension of .DUD.  The contaminant data files store
       the user-specified data, including contaminant names, paniculate-associated concentrations of
       contaminants  in  the  bulk  sediment, sediment solids  concentration,  standard   elutriate
       concentrations   of  contaminants,   water   quality  standards  for  the  contaminants,  and
       concentrations of contaminants in the background water at the disposal site.

g.     Input Data File Selection Menu

       Selecting F3 from the Evaluation  Selection Menu brings  up the  Input Data File  Selection
       Menu.  An input data file needs to be selected only when the user wants to edit data that were
       previously entered.  The changes can  be saved to the same  file or to a new file.  The first
       option  is used to  specify the name of the file  to be used.  The file specified in this option
       becomes the active data file.  If needed, the second option  is  used to specify the DOS path to
       the location where the data file should be read.  If a path  is not specified, the program will
       use  the default directory, where the STFATE program is,  for file  storage.  The third option
       displays a directory of STFATE input data files for the current path, that is files having an
       extension of .DUI in the directory  specified in the path.  An existing data file name may be
       selected from the list to use as the active data  file name for  reading existing data.  After the
                                                                                 DRAFT

-------
                                              C-27

       input file has been selected, press F4 to build or edit the input data file.  The input data that
       are stored in the selected  file  are then read  and  will  later be displayed on the input data
       screens to be reviewed and edited.  If the specified file could not be found (did not exist), the
       program will provide the user  the opportunity to  initialize  the file and start creating a new
       data set.

h.     Disposal Operation Selection Menu

       Selecting F4 from the Input Data File Selection Menu brings up this menu. The selection of a
       disposal type under this menu controls the input data requests, the type of execution data file
       that will be  built,  and the open-water disposal model that will  be executed.   Select the
       appropriate type of disposal:  Fl -  Disposal  from a Multiple Bin Hopper Dredge, or  F2 -
       Disposal from a Split-Hull Barge or  Scow.  The STFATE Input Selection Menu will then be
       displayed.

i.      Input Selection Menu

       Five types of input data have to be entered as shown in Table C-2 and Figure C-4, plus any
       desired changes in the default set of model coefficients, before an  execution data  file  can be
       written.  Default values are included for all of the model coefficients requested.  Enter data
       by paging down through the data entry screens, making selections and filling in the cells for
       each option.  An input data file  may be written at any point to save all the data that have been
       entered up  to that  point.   After entering all of  the data,  the data must be saved  before
       returning to the STFATE Activity Selection Menu  to avoid losing  the changes.  The data are
       saved in input data and execution data  files by selecting F7 from the Input Selection Menu to
       bring up the Input File Saving Menu.

j.      Input File Saving Menu

       This menu provides the opportunity to write an input data file to save the input data for future
       editing  under the STFATE  Input Selection Menu  and an execution data file for  use  during
       execution of the STFATE model. Execution data files are the actual  input data files used by
       the open-water disposal model to perform the analysis  and  generate output.   These files are
       unique  in structure to  the input requirements of a particular open-water disposal operation and
       contain data only for the specific options selected in the input.  The files are stored with the
       same name  as the input data file but with an extension of .DUE instead of .DUI.  The input
       data file stores data for all possible options in evaluations, disposal operations and methods of
       data entry,  allowing the user to perform comparisons between options  without  re-entering
                                                                                DRAFT

-------
                                             C-28

       previously specified data.  This menu is similar to the Input Data Selection Menu (see step g).
       The only  difference  is that the name of the file to be saved instead of read should be
       specified.  The same file as read can be used but the input and execution  data files will be
       overwritten.  If the input data are complete, an execution data file will also be saved.  After
       the files are  saved, the program returns control to the Input Selection Menu.  At this point
       data entry is  complete and the user should return to the Activity Selection Menu by hitting the
       Esc key.

k.     Execute

       Selecting F2  from the Activity  Selection Menu (see step b) initiates execution by bringing up
       the Execution  Data  File Selection  Menu.    After selecting  the  execution data  file (same
       procedure as the  other file selection  menus),  pressing F4  begins the simulation.  This option
       uses the execution data file to generate an output file and three graphics  files of the same
       name as the  execution data file selected but with an extension of .DUO, .DUP, .DUG, and
       .DUT,  respectively,  instead of .DUE.  The  execution may take a few minutes or several
       hours, depending on the simulation selected and the computer hardware used, but typically 30
       minutes is sufficient.  After termination of the simulation  the program returns to the Activity
       Selection Menu.

1.      Print or View Output

       Select F3 from the Activity Selection Menu to print or view text output. A STFATE Output
       Data File Selection Menu will  be displayed that  is similar to the other file selection menus.
       The output files have the same name as the execution data files used to generate them except
       that they have a  .DUO extension instead of a .DUE extension.  The  other difference in the
       menu is that it has an option to view the output  on the monitor using the LIST.COM utility
       program.  Instructions on using the LIST program are provided on the menu bar and on-line
       by pressing the ? key.  The output  is an ASCII  text file  having 132 characters per line and
       should be printed using compressed print or wide paper.   The program will  automatically use
       compressed print on some printers, mainly Epson and IBM printers.  It may be necessary to
       turn on compressed printing on your printer prior to printing the output, or to print the output
       outside the STFATE program, using the DOS print  command or a  word processor.  The
       output  contains an interpretive listing of the input data, computational  indicators, convective
       descent  results,  collapse results, information on cloud  generation  for transport-diffusion
       simulation,  accumulation  and  thickness of deposited  materials,  spatial distribution  of
       concentrations of materials in the water column, and water quality comparisons with standards
       for determining water quality violations or mixing zone requirements.
                                                                                DRAFT

-------
                                             C-29

m.     Generate Graphics

       Selecting  F4 from the Activity Selection Menu brings up the Graphics File Selection Menu
       which is similar to the other file selection menus.  Unlike the other file selection procedures,
       there are three graphics files, not one as for input, execution and output. All three files have
       the same  name as the execution file used to generate the output and graphics files, but have
       different extensions.  The graphics file selection procedure uses the graphics  file with an
       extension of  .DUT for its  directory listing and  file  searching.   Selecting F4 (Generate
       Graphics  with Selected  File) brings up  the Graphics Generation Menu from which there are
       three options  for plotting the data, one for each of three types of graphics  files.  The three
       options are Fl  -  Maximum Concentrations  Versus  Time,  using the file with a  .DUP
       extension; F2 -  Concentration Contours in Horizontal Plane, using the file with a  .DUC
       extension; and F3 -  Deposition Thickness at End of Simulation, using the file with a  .DUT
       extension.  The plots can be viewed on the monitor, or sent to a printer or plotter as desired.
       However, before plots  can be generated the software must be configured for the  hardware
       present.

n.     Perform Hardware Configuration for Graphics

       Select F5 from the Activity  Selection Menu to choose the proper printer, plotter and video
       information for the computer system being used.

o.     Ending

       To exit the program, press Esc repeatedly until you obtain a DOS prompt.  During execution
       of  a  particular  application's program,  the user has to wait  until the sometimes lengthy
       computations are computed.  The  program can also be terminated by a Control-Break  which
       will stop the execution after the next screen update and provide partial output. Alternatively,
       the program can  be  stopped  by turning off or  rebooting the  computer, but loss of data and
       output will  occur.  These methods of ending  are not recommended.   Similar  methods are
       available during printing of output.
C2.8          STFATE Application Examples

Two example applications of the use  of the numerical  model STFATE are described.  The first
example  addresses the instantaneous disposal of dredged material  from a split-hull barge or  scow.
These barges or scows may hold anywhere from approximately 400  to 6000 yd3 of material  and
                                                                               DRAFT

-------
                                            C-30

dispose of the material by means of opening the split-hull and discharging the material through the
bottom opening.  The material then  descends through the water  column to the bottom of the water
body.  The second example illustrates the modeling of dredged material disposal from  a multiple-bin
hopper dredge.  A  hopper dredge fills its bins with dredged material and then transports it to the
disposal site where  it discharges the material.  Each bin has  a separate opening in the  ship's bottom
through which the  dredged  material is  discharged  into the water  column.  Typically there are
anywhere  from about 4 to 20 bins hi a hopper dredge which  can  carry a total of approximately  1000
to 9000 yd3 of material.  During disposal one or more bins are  opened sequentially until all of the
bins have  been emptied.  The required input data for both examples are described and the results or
output from the STFATE model are illustrated and discussed. Additionally, the input and output files
for each of the examples  are included on diskettes which are  located in the pocket at the back of this
manual  along with  the  STFATE module  of the  Automated  Dredging  and Disposal Alternatives
Management System (ADDAMS).
C2.8.1        Split-hull Barge or Scow Example

An example of dredged material disposal is modeled for an instantaneous disposal using STFATE for
a 3000  yd3 disposal from a split-hull barge at a constant  40 ft  depth site for Section  404(b)(l)
regulatory analysis for water quality.  The input data for this example are given in Table  C-3.  No
mixing zone dimensions  are  specified for this example, therefore the dimensions of a mixing zone
required  to  meet the water quality  standard are calculated.  A description follows for entering the
required  example data and the use of the STFATE module.
C2.8.1.1       Entering STFATE and the Input Data File Selection Menu

The STFATE model is executed from the disk operating system (DOS)  prompt and the "STFATE
Activity Selection Menu" is reached as presented earlier.  The menus are  shown in Figs. C-3 and C-
4.  To proceed, the "Build or edit input data file"  option is selected and  the "STFATE - Short-term
Fate of a Disposal in Open Water Evaluation Selection Menu" appears.  For this example, the option
"Section 404(b)(l) Reg. Analysis for Navigable Waters" is selected.  Next, the "STFATE Input Data
File Selection Menu"  is presented and the key Fl  is pressed to  enter name  of input data file to be
built or edited.  For this example, BARGE is typed  and the ENTER key is pressed.  Option F4,
"Build or edit input data file," is then selected to read the input data file if it exists or to initialize it if
it is a new file.  A descriptive tide, "Barge dump  without specified mixing zone (Tier II W.Q.)"  is
typed and entered (Press ENTER).  The  "Disposal Operation Selection  Menu" is presented next.
"Disposal from a Split-Hull Barge or Scow" is  selected and the "STFATE Input Selection Menu"
                                                                               DRAFT

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                                       C-31

Table C-3.     STFATE Input Variables for Section 404(b)(l)  Regulatory  Analysis  of Navigable
             Waters Using a Scow/Barge Disposal.


INPUT PARAMETER                UNITS                   INPUT VALUED
SITE DESCRIPTION
Number of grid points (L-R. +z dir)
Number of grid points (T-B. +x dir)
Grid spacing (L-R). f(V)
Grid spacing (T-B)r f(V)
Constant water depth
Bottom roughness
Bottom slope fa-dirt
Bottom slope (z-dir)
Number of points in density profile
Density at point one (surface)
Density at point two (bottom)
VELOCITY
Type of velocity profile
Water depth 2-point profile
Vel for 2-point x-direction
Vel for 2-point z-direction
INPUT. EXECUTION & OUTPUT
Process to simulate
Duration of simulation
Time step for diffusion. f(V)
Convective descent output
Collapse phase output option
Number of print times for diffusion
Upper left corner mixing zone
Lower right corner mixing zone
Contaminant
mgyi
Predicted initial concentration in fluid
Background concentration
Number of depths for output
Depths for output


ft .
ft
ft
ft
deg
deg

g/cc
g/cc

ft
ft/s
ft/s
KEYS

s
s



ft
ft
Lead
0.0032
mg/L
mg/L

ft
32
32
50
200
40
0.005
0
0
2
1.0000
1.0002
2pt
30.38
0.5. 0.3
0.0
Disp. from Split-Hull Barge/Scow
3600
300
Yes
Yes
Quarterly
0.0
0.0
WO standard at edge of mixing zone

0.174
0.0002
2
15.39
                                        (Continued)
                                                                     DRAFT

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                                           C-32
Table C-3 (continued).
STFATE Input  Variables for  Section 404(b)(l) Regulatory Analysis  of
Navigable Waters Using a Scow/Barge Disposal.
INPUT PARAMETER UNITS
MATERIAL DESCRIPTION
Number of solids fraction
Solid fraction descriptions
Solid fraction specific gravity
Solid fraction volume concentration vd3/vd3
Solid fraction fall velocity ft/s
Solid fraction depositional void ratio
Solid fraction critical shear stress lb/ft2
Cohesive (Y/N)
Stripped during descentfYYN)
Dredge site water density g/cc
Number of lavers
Volume of each laver vd3
Vessel velocity in x-direction ft/s
Vessel velocity in z-direction ft/s
DISPOSAL OPERATION
Disposal point top of grid ft
Disposal point left edge of grid ft
Length of vessel bin ft
Width of vessel bin ft
Bottom depression length x-direction ft
Bottom depression length z-direction ft
Bottom depression average depth ft
Predisposal draft ft
Postdisposal draft ft
Time to emptv vessel s
COEFFICIENTS
Settling coef (BETA)
Apparent mass coefficient (CM)
Drag coefficient (CD)
Form drag collapse cloud (CDRAG)
Skin friction collapse cloud (CFRIC)
Drag ellipse wedge (CDS')
Drag plate (CD4)
Friction between cloud and bottom (FRICTN)
4/3 Law horizontal diffusion coefficient (ALAMDA^
Unstratified vertical diffusion coefficient (AKYO)
Cloud/ambient density gradient ratio (GAM A)
Turbulent thermal entrainment (ALPHAO)
Entrainment collapse (ALPHAQ
Stripping factor (CSTRIP)
INPUT VALUE
3
clumps, sand clay
1.6. 2.7. 2.65
0.1.0.2.0.05/0.0.0.15.0.10
3.0.0.1.0.002
0.4. 0.6. 5.0
99. 0.025. 0.002
N. N. Y
N. Y. Y
1.0
2
2000 / 1000
6/6
0/0
1300
750
200
50
0
0
0
17
5
20
0.0
1.0
0.5
1.0
0.01
0.1
1.0
0.01
0.001
0.025
0.25
0.235
0.1
0.003
(Concluded)

                                                                            DRAFT

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                                             C-33

appears on the computer monitor.  Now the entering of the input given in Table C-3, developed from
Table C-2, begins.
C2.8.1.1.1        Site Description Data

"Fl - Site Description" is selected from the "STFATE Input Selection Menu" by pressing key Fl or
by using arrow keys to highlight the selection and pressing ENTER.  The number of grid points  is
selected as 32 in both  the x-
-------
                                               C-34
                                   on
                             arid PI. 1
                        o it  and PI. i
                                                                   ISSOtt
                                                              Orld PI. 32
                      4660 It Grid Pt. 32
                                      1 —
                                      1
                                                  760 «
                                         low n
                                                  an/*
                                                        W«Ur
                                                       Currant
                                                     (0 to 0.6 R/«)
                                            W«Ur Depth - 40 ft
OrM
Boundary
Figure C-5.        Schematic of Example Disposal Site for Barge Disposal.
                                                Velocity (It/*)
                                               0.0     0.3   0,6
                                Depth (ft)
Figure C-6.        X-direction Velocity Profile for Barge Example.
                                                                                    DRAFT

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                                            C-35

C2.8.1.1 J        Input, Execution and Output Keys

At this point,  "F3-Input, Execution and Output Keys" is  selected in the same method as previously
described  which brings  the  "Simulation Selection  Menu" to the monitor.   Since initial  mixing
calculations are desired, "F3-DESCENT, COLLAPSE AND LONG-TERM DIFFUSION" is selected.
Next, the "Evaluation Selection  Menu "appears on the monitor,  and  since this  example  requires
comparison  to water  quality  concentrations  for  lead,  the "F2-TIER  II, COMPARE  WATER
QUALITY"  is chosen.  This  selection also provides for calculation of the size (length and width) of
the mixing zone required to prevent violation of a specified water quality standard.  When calculation
of the mixing  zone is desired, zeros are  entered in the boxes for the upper and lower  mixing zone
corners.  In this example, the  contaminant of concern is lead which has a background concentration of
0.0002 mg/L.   The predicted initial concentration in the fluid fraction is 0.174 mg/L and the water
quality standard at border or mixing  zone is  0.0032  mg/L.  After data are entered, press PAGE
DOWN to receive  a screen which asks if a zone of initial dilution is desired.  For this example, the
answer NO is  selected using arrow keys.  Press PAGE DOWN to specify the depths at which water
quality results are  desired.  The number of depths, between 2 and 5,  and the corresponding depth
value are entered.  In  this example,  two depths of 15 and  39 ft are entered.  Using PAGE DOWN, a
data  entry screen for the duration of simulation, long term time step, and specifications for printed
output are displayed.  Since the  example is for water column evaluation, a one hour (3600 s)  duration
and  a time step of 300  s is  input.  Output concerning  convective descent and collapse phase are
requested to permit review of the simulation.  Particular times are not of importance for an example,
so NO is chosen and the output is produced quarterly (900, 1800, 2700 and 3600 s).  Specific times
can be requested by choosing YES  and another screen will appear  for  entering these times,  but the
times must be  increments of the selected time step.  Again, PAGE DOWN is pressed  returning to the
"STFATE Input Selection Menu."
C2.8.1.1.4        Material Description Data

The material description data are entered after choosing the "F4-Material Description Data" from the
"STFATE Input Selection Menu."  Next, the first material description data entry screen appears  and
requests information on the number of layers (1 to 6) of material in the barge and the total volume of
each layer.  In the example, the number of layers is 2 and their volumes are 2000 and  1000 yd3,
respectively.  Press PAGE DOWN to get the next screen which provides for specifying the barge
velocity in terms of x- and z-direction components for each layer.  The  barge velocity is assumed to
be constant at 6 ft/s in the x-direction.  After pressing PAGE DOWN, material separation in the
barge is selected as YES for this example; that is, the concentration of solids vary from layer to layer
in the discharges from the barge.   Also  requested is the number of solid fractions (1  to 4) in the
                                                                              DRAFT

-------
                                             C-36

material, such as clumps, gravel,  sand, silt or clay; this example uses 3 solid  fractions.  The next
screen inputs the physical characteristics of the solids fractions which are entered in the highlighted
boxes on the screen.  Typical values and their ranges are shown at the top of  the screen.   For the
example, the input values are shown in Table C-3. Press PAGE DOWN to get the next screen which
asks if the  adjustment  of the entrainment  and  drag coefficients based on the moisture content is
desired.  Typically, this  is not necessary and NO is selected as was done for the example.  The final
screen for material description requests input on the density of the dredging site water which is in the
barge with the dredged material solids.  For the example, the density is entered directly (YES to first
question) and the value of 1.000 g/cc is accepted.  If the density is different, it can be entered in the
highlighted box.  If salinity and temperature data only are available, then NO is selected and another
screen will appear to allow for the input of those data.  At this point, PAGE DOWN is pressed which
brings back  the "STFATE Input Selection Menu."
C2.8.1.1.5        Disposal Operation Data

To describe the disposal operation, "PS-Disposal Operation Data" is selected which brings up a screen
requesting input concerning location of disposal point,  length and width of barge bin, the barge draft
before and after disposal, and the time needed to empty the barge.  The actual data are entered into
the respective highlighted boxes on  the screen.  For this example the location of the disposal point in
distance from the top of grid of 1000 ft and from the left edge of grid of 750 ft are entered.  The
length and  width of barge bin are entered as 200 and SO ft,  respectively.  The pre- and post-disposal
draft are entered as 17 ft and 5 ft, and the time to empty barge is 20 s.  Pressing PAGE DOWN gets
the next screen  which requests information  concerning disposal in a pre-existing depression.  For the
example no depression is used and  the values of zero  are accepted.  PAGE DOWN is pressed again
completing data entry and returning to  the "STFATE Input Selection Menu."
C2.8.1.1.6         Coefficients

The "STFATE Input Selection Menu" is now displayed on the monitor and the next selection is "F6-
Coefficients (Default Values)".  Highlighting  this selection and pressing  ENTER or pressing F6
shows  the numerical model coefficients.  In most cases in the absence of calibration data, the default
values  should  be chosen, and this is done in the example by pressing PAGE DOWN.  If other values
are required, then enter them in the highlighted box before pressing PAGE DOWN.  Expert guidance
should be obtained before using coefficient values other than the default numbers.
                                                                                DRAFT

-------
                                             C-37

C2.8.1.1.7        Saving Input Data Menu

Data entry is now complete as indicated by asterisks by each data entry option.  The next step is to
save the input data file for  use in the execution of STFATE.  To proceed, press the F7 key or select
"FT-Saving Input Data Menu" from the "STFATE  Input Selection Menu"  and press ENTER.  The
"Saving Input Data Menu"  appears and requests input as to whether a new file name is desired or the
active data file should be used for  storing the data. The file name entered at the beginning of the
input process appears as the active data file.  To save the data or changes in the active data file, select
option "F4-Save data in (or to) the active data file".  For this example, the active data file BARGE is
selected.  If the active data file exists, the program indicates the active  data file already exists and
requests to overwrite the file.  Therefore,  "Y" is  entered to overwrite and the program then requests
the entering or editing of  a descriptive title for  the file.   For this example the title "Barge  dump
without specified mixing zone (Tier II W.Q.)"  is entered.  Sometimes a file is being edited but the
original data  file needs to be kept unchanged.  At this point a new file  name can be selected using
option "Fl-Enter name of file to be saved"; the changes are then saved using option F4. After the
data are saved,  the  "STFATE  Input Selection Menu"  reappears  and all of the selections  show  an
asterisk indicating each selection has been completed.  The final step of the input process is to press
ESC and  return to the  "SFTATE Activity Menu"  which is the screen at which the input process
began.  Once the input  file is complete  and  saved, the STFATE model  can be executed by first
selecting "F2-Execute STFATE" which then requests the input data file to be input.  BARGE is input
to obtain results for this example which are discussed in the next section.
C2.8.1.2   Description of Barge Disposal Example Output

A general description of the output available from the STFATE has been described.  The objective in
this section is to illustrate  and describe selected portions of the results.  The results concerning the
accumulation of sediment on the bottom are not discussed since the emphasis is on results for water
column concentrations for Tier II  and III  evaluations.   However, bottom sediment  accumulation
output is contained in the BARGE.DUO file on the disk enclosed with this manual.   Output related to
the convective descent and collapse phase is also contained in the output file for the barge example
(BARGE.DUO)  for information purposes.  The results discussed are water  column concentrations of
the solids fractions, contaminant and the fluid associated with the dredged material in the barge.

Results are in the form of  tabular output and graphical presentations which can be  accessed through
the "STFATE Activity Selection Menu".   This screen provides two options for output, "F3-Print or
view output" and "F4-Generate graphics."  If option F3 is selected, the "STFATE Output Data File
Selection Menu"  appears, and it provides  several possibilities.  First, the option "Fl-Enter name of
                                                                               DRAFT

-------
                                             C-38

data file used during execution" is used  and for this example,  the filename BARGE  is entered after
selecting this option.  Once the filename is identified, the options "F4-Print selected output file"  or
"FS-View selected output file" are used.  There is considerable model output and all of the output is
usually  not desired.  The FS option provides viewing of the output file by paging through it using the
PAGE UP and PAGE DOWN keys.  The viewing software can also be used to select specific portions
of the output for printing selected hard copy.   Pressing the ESC key returns the "STFATE Activity
Selection Menu".
C2.8.1.2.1        Barge Disposal Water Column Concentrations and Area Distribution

In this  example,  water column concentrations  of  the solids fraction and the  contaminant were
requested at  15  and 39 ft.  Thus, the concentrations for clumps, sand, clay and  lead at every grid
point location for both depths are contained  in  the output file.    In addition, results are  available
showing the maximum concentration occurring at each  grid point  over the depth of the water column
for the duration of the simulation.   The  top  section  of Figure C-7 shows the  output  for lead
concentration in mg/L at 15 ft at the end of the model run (3600 s). The concentration values at each
grid point are must  be multiplied by the appropriate scale factor,  shown at top of the table, to get
actual concentration.  In this example the maximum value on the grid is 1.6 which  is multiplied by
0.001 yielding a maximum lead concentration of  0.0016 mg/L at x- and z-grid locations of 20 and
16, respectively. Recall from the input that the distance between  x-grid points is  150 ft and between
z-grid points is  50 ft.   Therefore, the maximum concentration occurs 2850 ft from top  of grid and
750 ft from left  edge of grid.  The disposal began at an x- and z-distance of 1000 ft (grid point 8) and
750  ft (grid point 16).   Both the barge and water velocity were  in the positive  x-direction so it is
reasonable to find  the  maximum concentration  down grid  from the disposal  point.    The lead
concentration area at 15 ft is evaluated by determining the number of  grid rectangles which have  a
value representing lead concentration and multiplying it by the area of the grid rectangle  (50 x  1500
= 7500 ft2).  A total of 129 grid points have a lead concentration above background of at  least 10~5
mg/L,  an area of 9.675 x 10s ft2.  Since the barge and the water current are moving in the positive x-
direction, it  is  expected that the distance in the x-direction should be longer than that in the z-
direction.  The  display of this area in Figure C-7 appears to be wider than it is long,  but that is
because of the grid spacing. It is actually 1000 ft wide and 1200 ft long (x-direction).
C2.8.1.2.2        Barge Disposal Water Column Concentrations

The water column concentrations over the duration of the simulation are tabulated in  the  middle
sections of Figure C-7.  This shows the clay and lead concentrations. The clumps and sand settled to
                                                                                DRAFT

-------
                                            C-39
CONCENTRATIONS ABOVE BACKGROUND OF LEAD MG/U X THE CLOUD 3800 00 SECONDS AFTER DUMP

         16.00 FT BELOW THE WATER SURFACE
...MULTIPLY DISPLAYED VALUES BY 0.1000E-02     (LEGEND... + . XT. .01 . . IT .0001 0 - .LT. .000001)
MN. 2
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12 OOOO
13 OOOO
14 OOOO

18 OOOO
17 OOOO
18 OOOO
1*0000
20 OOOO
21 OOOO
22 OOOO
23 OOOO

24 OOOO
26 OOOO
28 OOOO
27 OOOO
28 OOOO
28 OOOO
30 OOOO
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.23
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0
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0
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31
ooooo
OOOO
OOOO
OOOO

OOOO
OOOO
OOOO
OOOO
OOOO
OOOO
OOOO
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      SUMMARY OF CONCENTRATIONS FOR CLAY
                  MAX CONC ABOVE
                   BACKGROUND
                                GR
                                  (FT)
TIME
(HR)
0.25
0.50
0.75
1.00
0.25
0.50
0.75
1.00
DEPTH
(FT)
15.0
15.0
15.0
15.0
39.0
39.0
39.0
39.0
ON ENTIRE
(MG/L)
0.306E + 03
0.473E + 03
0.318E + 03
0.293E + 03
0.430E + 04
0.815E + 03
0.427E + 03
0.544E + 02
X-LOC
) (FT)
1500.
1950.
2400.
2400.
1350.
1650.
1950.
2400.
Z-

750.
750.
750.
750.
750.
750.
750.
750.
                              Z-LOC
      SUMMARY OF CONCENTRATIONS FOR  LEAD
                  MAX CONC ABOVE
                   BACKGROUND    MAX CONC
      TIME    DEPTH   ON ENTIRE GRID    ON GRID
      (HR)    (FT)     (MG/L)
      0.08    15.0    0.286E-12
      0.17    15.0    0.212E-02
      0.25    15.0    0.604E-02
      0.33    15.0    0.511E-02
      0.42    15.0    0.435E-02
      0.50    15.0    0.374E-02
      0.58    15.0    0.323E-02
      0.67    15.0    0.281E-02

Figure C-7.
                                 X-LOC   Z-LOC
                (MG/L)    (FT)    (FT)
                 0.200E-03   1200.    750.
                             1350.    750.
                             1500.    750.
                             1650.    750.
                             1800.    750.
                             1950.    750.
                             2100.    750.
                             2250.    750.
0.232E-02
0.624E-02
0.531E-02
0.455E-02
0.394E-02
0.343E-02
0.301E-02
Selected Output for Barge Disposal.
                                                                              DRAFT

-------
C-40
0.75
0.83
0.92
1.00
0.08
0.17
0.25
0.33
0.42
0.50
0.58
0.67
0.75
0.83
0.92
1.00
0.08
0.17
0.25
0.33
0.42
0.50
0.58
0.67
0.75
0.83
0.92
1.00
0.08
0.17
0.25
0.33
0.42
0.50
0.58
0.67
0.75
0.83
0.92
1.00
0.08
0.17
0.25
0.33
0.42
0.50
0.58
0.67
0.75
0.83
0.92
1.00
Figure C-7.
15.0
15.0
15.0
15.0
39.0
39.0
39.0
39.0
39.0
39.0
39.0
39.0
39.0
39.0
39.0
39.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
10.0
10.0
10.0
10.0
10.0
10.0
10.0
10.0
10.0
10.0
10.0
10.0
20.0
20.0
20.0
20.0
20.0
20.0
20.0
20.0
20.0
20.0
20.0
20.0
Selected
0.246E-02
0.216E-02
0.191E-02
0.169E-02
0.103E-01
0.231E-02
0.112E-02
0.950E-03
0.810E-03
0.695E-03
0.601 E-03
0.522E-03
0.457E-03
0.401 E-03
0.355E-03
0.315E-03
0.524E-02
0.135E-02
0.185E-02
0.156E-02
0.133E-02
0.114E-02
0.988E-03
0.859E-03
0.752E-03
0.661 E-03
0.584E-03
0.518E-03
0.237E-01
0.695E-02
0.421E-02
0.357E-02
0.304E-02
0.261E-02
0.226E-02
0.196E-02
0.171E-02
0.151E-02
0.133E-02
0.118E-02
0.393E-01
0.126E-01
0.683E-02
0.578E-02
0.492E-02
0.423E-02
0.365E-02
0.318E-02
0.278E-02
0.244E-02
0.216E-02
0.191E-02
0.266E-02
0.236E-02
0.211E-02
0.189E-02
0.105E-01
0.251E-02
0.132E-02
0.115E-02
0.101E-02
0.895E-03
0.801 E-03
0.722E-03
0.657E-03
0.601 E-03
0.555E-03
0.515E-03
0.544E-02
0.155E-02
0.205E-02
0.176E-02
0.153E-02
0.134E-02
0.119E-02
0.106E-02
0.952E-03
0.861 E-03
0.784E-03
0.718E-03
0.239E-01
0.715E-02
0.441E-02
0.377E-02
0.324E-02
0.281E-02
0.246E-02
0.216E-02
0.191E-02
0.171E-02
0.153E-02
0.138E-02
0.395E-01
0.128E-01
0.703E-02
0.598E-02
0.512E-02
0.443E-02
0.385E-02
0.338E-02
0.298E-02
0.264E-02
0.236E-02
0.211E-02
Output for Barge Disposal.
2400.
2550.
2700.
2850.
1200.
1350.
1500.
1650.
1800.
1950.
2100.
2250.
2400.
2550.
2700.
2850.
1200.
1350.
1500.
1650.
1800.
1950.
2100.
2250.
2400.
2550.
2700.
2850.
1200.
1350.
1500.
1650.
1800.
1950.
2100.
2250.
2400.
2550.
2700.
2850.
1200.
1350.
1500.
1650.
1800.
1950.
2100.
2250.
2400.
2550.
2700.
2850.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
(continued)
                          DRAFT

-------
                                            C-41
      0.08
      0.17
      0.25
      0.33
      0.42
      0.50
      0.58
      0.67
      0.75
      0.83
      0.92
      1.00

      0.08
      0.17
      0.25
      0.33
      0.42
      0.50
      0.58
      0.67
      0.75
      0.83
      0.92
      1.00
30.0
30.0
30.0
30.0
30.0
30.0
30.0
30.0
30.0
30.0
30.0
30.0

40.0
40.0
40.0
40.0
40.0
40.0
40.0
40.0
40.0
40.0
40.0
40.0
0.239E-01
0.794E-02
0.414E-02
0.350E-02
0.299E-02
0.256E-02
0.221E-02
0.193E-02
0.168E-02
0.148E-02
0.131E-02
0.116E-02

0.531E-02
0.172E-02
0.924E-03
0.781E-03
0.666E-03
0.572E-03
0.494E-03
0.430E-03
0.376E-03
0.330E-03
0.292E-03
0.259E-03
0.241 E-01
0.814E-02
0.434E-02
0.370E-02
0.319E-02
0.276E-02
0.241E-02
0.213E-02
0.188E-02
0.168E-02
0.151E-02
0.136E-02

0.551E-02
0.192E-02
0.112E-02
0.981E-03
0.866E-03
0.772E-03
0.694E-03
0.630E-03
0.576E-03
0.530E-03
0.492E-03
0.459E-03
1200.
1350.
1500.
1650.
1800.
1950.
2100.
2250.
2400.
2550.
2700.
2850.

1200.
1350.
1500.
1650.
1800.
1950.
2100.
2250.
2400.
2550.
2700.
2850.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.

750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
ESTIMATES OF AREAS CURRENTLY IN VIOLATION (SNAPSHOT) AND MIXING ZONES (ACCUMULATED AREA OF VIOLATION)
TIME
( SEC )
300.0
600.0
900.0
1200.0
1500.0
800.0
2100.0
2400.0
2700.0
3000.0
3300.0
3600.0
SNAPSHOT
ARE A (SO FT)
0.975000E+05
0.600000E+05
0.450000E+05
0.450000E+05
0.375000E+05
0.225000E+05
0.225000E+05
0.750000E+04
O.OOOOOOE+00
O.OOOOOOE+00
O.OOOOOOE+00
O.OOOOOOE+00
L(FT)
570.
391.
391.
391.
292.
212.
212.
158.
0.
0.
0.
0.
W(FT)
171.
154.
115.
115.
129.
106.
106.
47.
0.
0.
0.
0.
ACCUMULATED
AREA(SQ FT)
0.975000E+05
0.112500E+06
0.150000E+06
0.187500E+06
0.225000E+06
0.247500E+06
0.270000E+06
0.277500E+06
0.277500E+06
0.277500E+06
0.277500E+06
0.277500E+06
L(FT)
570.
570.
695.
828.
966.
1107.
1250.
1395.
1395.
1395.
1395.
1395.
W(FT)
171.
197.
216.
227.
233.
224.
216.
199.
199.
199.
199.
199.
Figure C-7.  Selected Output for Barge Disposal (concluded).
                                                                              DRAFT

-------
                                             C-42

the bottom before 900 s so there is no history for those sediment fractions.  Concentrations for clay
are shown to decrease with tune for both requested depths and the quarterly time entered in the input
data.  The lead concentrations are shown for  every time step for the two depths of IS and 39 ft as
well as depths 0, 10, 20, 30 and 40 ft.  The model evaluates concentrations at five additional depths
based on the location of clouds to better estimate the peak concentration of contaminant in the water
column.  The last section of Figure C-7 shows the snapshot mixing zone (at a instant in time)  and
accumulated mixing  zone (from the beginning of simulation).  The snapshot columns  show the area
that exceeds the water quality standard at the time of the results.  For example, at 300s a region 570
ft  long, 171  ft wide and 97,500 ft2 in  area has a lead concentration that  exceeds the water  quality
standard.  Areas were in violation up to and including 2400 seconds.  The accumulated  columns show
the area that exceeds or previously exceeded the standard at the time of the results.  The  appropriate
required mixing zone has an area of 277,500 ft2, a length of 1395 ft, and a width of 199 ft.
C2.8.1.2.3        Plots of Concentration Following Barge Disposal

From the "STFATE Activity Selection  Menu,"  "F4-Generate Graphics" is selected to receive the
"STFATE Graphics File Selection Menu" which has several options.  First,  the name of the data file
used during execution is entered after selecting "Fl-Enter name of data file used during execution".
Next, the "F4-Generate graphics  with selected  file" is  pressed to receive the "STFATE Graphics
Generation Menu".  The first option  is  to select "Fl-Maximum concentrations versus time" which
brings up a screen with selections for how the  graph is to be displayed (screen, printer, plotter),
which solids fraction or contaminant (clumps, sand, clay or lead in this example) is desired, and what
depth (15,  39 or peak) are desired.   Peak means  the water column  depths at which the maximum
concentrations  occur.   The  maximum concentration versus  time for lead in this  barge disposal
example (Fig.  C-8)  shows the maximum (0.039 mg/L)  occurring 5  min after disposal and rapidly
dropping to below the mixing  zone standard (0.0032 mg/L)  at  41  min.  It then stays under the
standard for the remainder of the simulation. Referring back to the middle sections of Figure C-7,  it
can be determined that the depth where the peak occurred is 20 ft.

Selecting the option  "F2-Concentration contours in horizontal plane" displays a screen which provides
the ability to graphically display the concentration contours of the contaminant or the solids fraction.
It also provides capability for graphically displaying the predicted mixing zone required.  As before,
the graphs can be output to  the screen,  plotter  or printer.  Concentration contours are obtained by
selecting the solid fraction or contaminant (clumps, sand, clay or lead) and then selecting the depth
desired (15, 39, or peak in this  example).  The user may select default contours (YES) or specify the
desired contours (NO).   Selecting NO  to  specify contours and  then pressing PAGE DOWN, the
desired contours are entered sequentially on the next screen.  The water quality standards will already
                                                                                DRAFT

-------
                                            C-43
              B.Btt
           C  8.832
           0
           N
           C  8.824
              9.616
              B.BBB
              fl.BOB
                      BARGE DUMP UITHOUI SPECIFIED MIXING ZONE (HER II U.Q.)
                                                                           IXAD
16     24     32     48
        TINE - Minutes
48
56
                                                                       M
                       *  MM CONG ON GRID
                                                — M.Z.
Figure C-8.        Peak Lead  Concentration in Water Column as a F(Time) for the Barge Disposal
                  Example.
be specified when the mixing zone, contaminant or fluid is selected for display.  Figure C-9 shows
contours of concentrations of lead above background at  a depth of IS ft after 3600 s.  The contour
values are  specified  for contours 1, 2  and  3 as 0.0032, 0.0001 and  0.00005 mg/L.   As shown,
contour 1 is not displayed in Figure C-9 indicating the concentration value of 0.0032  mg/L above
background which is the water quality standard entered in the input file is not exceeded on the  grid.
For this example, the  input requested  that the mixing zone be predicted.  Figure C-10 shows the
predicted "peak" mixing zone outside of which the 0.0032 mg/L standard  is not exceeded during the
simulation  at any depth.   To plot the predicted mixing  zone, press the F2  key from the "STFATE
Graphics Generation Menu" which displays  a new screen.   Now, select the box  "Mixing" and the
desired depth.   Press PAGE DOWN  to receive  the  next screen  and select  time and  whether
defaultcontours are used.   In this  example,  choose NO and press PAGE DOWN.   The following
screen requests user-specified  contour  values.  Insert contour values and  press PAGE DOWN to
receive graph.  This completes the example  and the ESC key is pressed repeatedly to return to the
"STFATE Activity Selection Menu" or to quit the program and return to DOS prompt.
                                                                              DRAFT

-------
                                           C-44
BMKE MMF UITHMIT SP
•
X 12H
D
I
C
T
I
0
" xae
4880
Kffin> HIXINB 2



I O )
ma am ii U.Q.) LEAD
NC/L
15.88 n
3688. SBC

CONTOUR UALUE8
sssssssssssssss
2» l!eB8E-83
1 1£M
2 1IHECTION
Figure C-9.        Lead Concentration Contours for 15 ft Depth at 3600 sec for the Barge Disposal
                  Example.
BARGE VJHf WITHOUT SPECIFIED MIXING ZONE (TIER II U.Q.) NIXING


X 1200
D
I
R
J Z40B
I
I
0
M 36BB
4MM


- o
w
V
V




nor b
PEAK FT
3&B8. SEC


COMTOUR VALUES
1 « 3.288E-83




                                    e          1680
                                     Z DIBECTION
Figure C-10.       Plot of Required Mixing Zone for the Barge Disposal Example.
                                                                             DRAFT

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                                             C-45

C2.8.2            Multiple-bin Hopper Dredge Example

An example of dredged material disposal is modeled using STFATE for a 3000 yd3 disposal from a
hopper dredge at a constant 40 ft depth site for Section 404(b)(l) regulatory analysis for water column
toxicity.   A  mixing zone  is specified  for this  example, therefore the calculated dredged  material
concentrations at  the boundary of the mixing zone are compared to the allowable concentrations as
determined by bioassay tests.  A description follows for entering the required example data and the
use of the STFATE module.


C2.8.2.1   Entering STFATE and the Input Data File Selection Menu

Many of the  steps and procedures for entering  the STFATE model for application to a multiple-bin
hopper disposal operation are the same as that previously described for the barge disposal  (Section
2.8.1.2).  Therefore some repetition is  contained in this  section  and a complete description  is given
for the purpose of clarity.

The  STFATE is  executed  from the DOS  prompt, and the "STFATE Activity Selection Menu" is
reached as described previously.  To proceed, the "Build or edit input data file" option  is selected.
This results in the  "STFATE -  Short-Term Fate of a Disposal in Open Water Evaluation Selection
Menu" being presented.  For this example, the option F3  for "Section 404(b)(l) Reg. Analysis for
Navigable Waters"  is selected.  As a result, the "STFATE Input  Data File Selection Menu" appears
and key Fl is pressed to enter the name of the input data file to be built or edited.  In this example,
HOPPER is typed  in the highlighted box  and  then the ENTER  key is pressed. Option F4 is then
selected to read the input data file if it exists  or to initialize it  if a new file.  A descriptive title,
"Example hopper dredge disposal with specified mixing zone (Tier III)," is entered (Press ENTER).
The  "Disposal Operation  Selection Menu"  appears  and  "Disposal  from  a  Multiple-Bin  Hopper
Dredge"  is selected (F2 key) which brings up the "STFATE Input Selection Menu."  Entering  of the
input data given in Table C-4 now begins.


C2.8.2.1.1        Site Description Data

From the "STFATE Input Selection Menu", "Fl-Site Description" is selected by pressing key  Fl  or
by using arrow keys to highlight selection and pressing ENTER.  On the first  data entry screen, the
number of grid points is selected  as 32 in both the x-direction (top to bottom) and z-direction Qeft to
right).  The spacing is picked as 50 ft in the z-direction and 150 ft in the x-direction.  These spacings
are selected because the water velocity, described later, is 0.5 ft/s  in the x-direction and 0.0 ft/s  in the
                                                                               DRAFT

-------
                                        C-46
Table C-4. STFATE Input Variables for Section 404(b)(l) Regulatory Analysis of Navigable Waters
          Using a Multiple-bin Hopper Dredge Disposal.
INPUT PARAMETER
UNITS
INPUT VALUE
SITE DESCRIPTION
Number of grid points (L-R. +z dirt
Number of grid points (T-B. +x dirt
Grid spacing (L-R). f(V)
Grid spacing (T-B) f(V)
Constant water depth
Bottom roughness
Bottom slope (x-dir)
Bottom slope (z-dir)
Number of points for density profile
Density at point one ("surface)
Density at point two (bottom)
VELOCITY
Type of velocity profile
Water depth of averaged velocity
Vertically averaged x-dir velocity
Vertically averaged z-dir velocity
INPUT, EXECUTION & OUTPUT
Process to simulate
Duration of simulation
Time step for diffusion. f(V)
Convective descent output
Collapse phase output option
Number of print times for diffusion
Upper left corner mixing zone
Lower right corner mixing zone
Toxicitv standard for dilution
Number of depths for output
Depths of transport-diffusion output
MATERIAL DESCRIPTION
Number of solids fraction
Solid fraction descriptions
Solid fraction specific gravity
Solid fraction volume concentration
Solid fraction fall velocity


ft
ft
ft
ft
deg
deg

g/cc
g/cc

ft
ft/s
ft/s
KEYS

s
s



ft
ft
%

ft



vd3/vd3
ft/s
32
32
50
200
40
0.005
0
0
2
1.0000
1.0002
1 pt. depth ave.. logarithmic
40
0.5
0
Disp. from Multiple-bin Hopper Dredge
3600
300
No
No
Quarterly
1000. 450
3000. 1050
0.4
2
20/39
3
sand. silt, clay
2.7. 2.65. 2.65
0.1.0.07.0.04
0.1.0.01.0.002
(Continued)
                                                                      DRAFT

-------
                                          C-47

Table C-4 (continued).     STFATE Input Variables for Section 404(b)(l) Regulatory Analysis of
                        Navigable Waters Using a Multiple-bin Hopper Dredge Disposal.
Solid fraction depositional void ratio
Solid fraction critical shear stress
Cohesive CYIN)
Stripped during descentfYVN)
redge site water density
INPUT PARAMETER
DISPOSAL OPERATION
Disposal point top of grid
Disposal point left edge of grid
Length of vessel bin
Width of vessel bin
Distance between bins
Time required to emptv all hopper bins
Number of bins opening simultaneously
Number of discrete openings of bins
Vessel velocity in x-dir for each set
Vessel velocitv in z-dir for each set
Bottom depression length x -direction
Bottom depression length z-direction
Bottom depression average depth
Predisposal draft
Postdisposal draft

Ib/ft2


g/cc
UNITS
ft
ft
ft
ft
ft
s


ft/s
ft/s
ft
ft
ft
ft
ft
0.6. 3.0. 5.0
0.025. 0.01. 0.002
N. N. Y
N. Y. Y
1.0
INPUT VALUE
1300
750
60
20
5
60
2
3
6/6/6
0/0/0
0
0
0
18
5
COEFFICIENTS
Settling coef (BETA)	0.0
Apparent mass coefficient (CM)	1.0
Drag coefficient (CD)	0.5
Form drag collapse cloud (CDRAG)	1.0
Skin friction collapse cloud (CFRIO	0.01
Drag ellipse wedge (CD3)	0.1
Drag plate (CD^	1.0
Friction between cloud and bottom (FRICTN)	0.01
4/3 Law horizontal diffusion coefficient (ALAMDA)	0.001
Unstratified vertical diffusion coefficient (AKYO)	0.025
Cloud/ambient density gradient ratio (GAMA)	0.25
Turbulent thermal  entrapment (ALPHAO)	0.235
Entrainment collapse fALPHAC)	0.1
Stripping factor (CSTRIP)	0.003
                                     (Concluded)
                                                                          DRAFT

-------
                                             C-48
z-direction.  Thus, the disposal site (Fig. C-ll) is  1550 ft wide and 4650 ft long.  A constant water
depth is entered as 40 ft, and the bottom roughness is input as the mid range value of 0.005.  A flat
bottom is assumed  so  a slope of zero is entered for the x- and z-directions.   Once the data entry
screen is complete, the PAGE DOWN key is pressed to move to the next screen.
                                • n
                           OrU Pt. 1
                      > *  OrU Pt 1
                                              1CCOH
                                          Grid Pt. 31
                    40EO It Qrld Pt. M
                r"
                                               750 H
                                           460 ft      10M ft
                                  *X
                                      1000 ft-
                             toW*
                                                       7WH
                                                       Zo««
                                                       Boundary
                                                       (0.4S)
                                                       1000 n
                                                   W«tw
                                                  Curr»it
                                          W«tti Dtpth - 40 n
                                                                   Grid
                                                                   Bouidtiy
Figure C-ll.      Schematic of Example Disposal Site for Multiple-bin Hopper Dredge Disposal.
C2.8.2.1.2
Velocity Data for Hopper Disposal Example
The selection of  "F2-Velocity Data"  from  the  "STFATE Input Selection  Menu" brings up  the
"Velocity Profile Selection Menu."  In this example a depth averaged water velocity profile (Fig. C-
12) for a constant depth is selected by pressing the F2 key or highlighting the selection using arrow
keys and then pressing PAGE DOWN. The next data entry screen appears on the monitor, and the
velocity and constant water depth data are entered in the highlighted boxes.  In this case, x-direction
velocity of 0.5 ft/s at a depth of 40 ft and z-direction velocity of 0 ft/s are entered. Although zero
velocity can be input, it is recommended that the speed of the resultant velocity vector be at least 0.1
ft/s because most open bodies of water have some motion occurring at all times.  When the input of
velocity data is complete, press PAGE DOWN to return to the "STFATE Input Selection Menu."
                                                                               DRAFT

-------
                                            C-49
                                        Velocity (ft/*)
                                       O.P
                                                  0.6
                            Depth (ft)
                                                       Logarithmic
                                                         Profile
Figure C-12.
X-direction Velocity Profile for Hopper Dredge Example.
C2.8.2.1.3
Input, Execution and Output Keys
"F3-Input,  Execution and Output  Keys" is selected as previously  described and  the  "Simulation
Selection Menu" appears.  Initial  mixing calculations are desired for the hopper disposal so "F3-
DESCENT, COLLAPSE  AND LONG-TERM  DIFFUSION"  is selected.   Next,  the  "Evaluation
Selection Menu" appears  on the monitor,  and  since this  example requires  comparison to  toxicity
results, the "F3-TIER III, COMPARE TOXICITY RESULTS"  is chosen.  The first input, execution
and  output keys data  entry  screen appears and  provides   for the  input  of the mixing zone
characteristics  and the toxicity standard for  dilution as  a percentage  of the initial  water  column
concentration prior to disposal.  For  this example, the mixing  zone upper left corner (x  = 750 ft, z
= 450 ft) and lower right corner (x = 3000 ft, z =  1050 ft) are entered and the dilution requirement
to meet the toxicity standard is appropriately  entered.  The percent of initial concentration is 0.4%.
After entering  the data, press PAGE  DOWN  to receive the next screen.  It asks  if a zone of initial
dilution is desired, and the answer  is  NO for this example.  Press PAGE DOWN  to receive the next
screen which requests the number of depths (2 to 5) and depths where  output on concentrations  are
desired.  In this example 2 depths, 20 and  39 ft,  are entered.  Using PAGE DOWN, the next data
entry screen requests  further input concerning the duration of simulation, long-term  time step and
specifications for printed output.   Since the example is for water column evaluation, the one hour
(3600 s)  duration and a  time step  of 300  s are input.   Output  concerning convective  descent and
collapse phase are not of particular interest for this example,  so NO  is selected.  Also, particular
                                                                              DRAFT

-------
                                             C-50

times for printing time diffusion results are not of importance since summary concentration data are
provided for all  time  steps,  so NO is chosen which  causes output to be produced quarterly (900,
1800, 2700 and 3600 s).  Again, PAGE DOWN is pressed and the "STFATE Input Selection Menu"
is returned.
C2.8.2.1.4        Material Description Data

The material description data are entered after choosing the "F4-Material Description Data" from the
"STFATE Input Selection Menu."  The next data entry screen appears and requests information  on
the total volume of dredged material in the hopper dredge (total of all bins) and the number of solids
fractions in the material. For this hopper disposal  example, the total volume  is 3000 yd3, and the
number of solid fractions is 3.   The next screen is used to enter the physical  characteristics of the
solids fractions which are entered in the highlighted boxes on the screen.  Typical values and their
ranges are shown at the top of the screen, and the input values are  shown at the bottom of the data
entry screen and are tabulated in Table C-4.  Press PAGE DOWN to get the next screen which asks if
the adjustment of the entrainment and drag  coefficients based on the moisture  content are desired.
Typically, this is not necessary and NO is selected as is done  in this example.   The final screen for
material description requests input on the density of the dredging site water which is in the hopper
with the dredged  material  solids.  For the  example, the density is entered directly (YES to first
question) and the value of 1.000  g/cc is accepted. If the density is different, it  can be entered  in the
highlighted  box.  If salinity and temperature  data are only  available, then NO is  selected and another
screen will appear to allow for the input of that data.  At this point, PAGE DOWN is selected and the
"STFATE Input Selection Menu" returns.
C2.8.2.1.5        Operation Data

To describe the disposal operation, "PS-Disposal Operation Data" is selected and the first data entry
screen is used to enter data concerning location of disposal point, length and width of disposal vessel
(hopper) bin, distance between bins,  pre- and post-disposal hopper draft, total time needed to empty
all bins and the number of bins that are opened simultaneously.  A schematic of the hopper dredge
bin layout is illustrated in Figure C-13.  The actual data are entered  into the respective highlighted
boxes, and the input data values are given in Table C-4.  For this example, the disposal point  is 1000
ft from the top edge of grid and 750 ft from left edge of grid (Fig.  C-ll).  The length and width of
each is 60 and 20 ft, respectively, and the  distance between the edge of bins is 5 ft.  The pre- and
post-disposal drafts are 18 and 5  ft,  respectively, and it takes 60 s  to empty all bins.  Press PAGE
DOWN to get next screen which requests information concerning the number of discrete openings of
pairs of hopper bins and the velocity of the hopper dredge.  For this example there are three discrete
openings of sets of two hopper bins.  Also, the hopper dredge is  assumed to travel at a constant
                                                                                DRAFT

-------
                                            C-51
                           SETS
                           SET 2
                           SET1
                                       FORWARD
                                       HOPPER
                                       DREDGE
Figure C-13.       Schematic of Hopper Dredge with 6 Bins. In this example, the bins are opened in
                  sets of two (bins 1 & 2, bins 3 & 4 and bins 5 & 6).

velocity of 6 ft/s during  all  three discrete openings.  After entering these data and pressing PAGE
DOWN, the next screen requires data entry concerning disposal in a pre-existing depression, and for
this example values of zero  are accepted.  PAGE  DOWN is pressed again and the  "STFATE Input
Selection Menu" reappears on the monitor.
C2.8.2.1.6
Coefficients
The  "STFATE Input Selection  Menu" is now on  the  monitor and  the  next  selection is  "F6-
Coefficients (Default Values)".   Highlighting this  selection  and pressing ENTER or pressing  F6
displays the numerical model coefficients.  In most cases the default values should be chosen and this
was done for the example by pressing PAGE DOWN.  If other values are required, they may be
entered in the appropriate highlighted box  before pressing PAGE DOWN.  Expert advice should be
obtained before using coefficient values different from the default numbers.
C2.8.2.1.7
Saving Input Data Menu
The entering of data is now complete as indicated by the asterisks by each data entry option.  The
next step is to  save the input data file for use in the execution of STFATE.  On the  "STFATE Input
                                                                             DRAFT

-------
                                             C-52

Selection Menu," press the F7 key or highlight  "F7-Saving Input Data Menu" and press ENTER.
The "Saving Input Data Menu" appears and requests input as to whether a new file name is desired or
the active data file should be used for storing the data.  The file name entered at the beginning of the
input process appears as the active data file.  Sometimes a file is being edited, but the original data
file needs to be kept unchanged.  At this point a new file name can be selected using option "Fl -Enter
name of file to be saved" and then option "F4-Save data in (or to) the active data file" is chosen to
save the data.  For the example,  the active data file HOPPER is selected.  If the active data file
exists, the program  indicates so and requests permission to overwrite the file.  In this example,  "Y" is
entered and the next screen requests a descriptive title for the file.  The title "Hopper Discharge with
specified mixing zone  (Tier III)" is entered.  Next, the  "STFATE Input Selection  Menu" reappears
and all of the selections show  an asterisk indicating each  selection has been completed.   To complete
the process, ESC is pressed  and the "STFATE Activity Menu"  appears.   Now  the input  file is
complete and saved, and the  STFATE  model can be executed by selecting  "F2-Execute STFATE"
which  then requests the input  data file to be entered.  HOPPER is entered to obtain results for this
example which  are discussed int the next section.
C2.8.2.2   Description of Example Hopper Disposal Output

The objective of this  section is  to  illustrate  and describe  selected portions  of the  hopper dredge
disposal results. Accumulation of sediment on the bottom is not discussed since the  emphasis  is on
results for water column  concentrations  for Tier II and III evaluations.  The results discussed are
concentrations of the solids fractions, contaminant and the fluid associated with the dredged material
in the hopper bins.

As previously explained in the barge example, results are accessed through the "STFATE Activity
Selection Menu".  This screen provides two options for output, "F3-Print or view output" and "F4-
Generate graphics."  If option  F3  is selected, the  "STFATE  Output Data  File  Selection Menu"
appears and provides several  possibilities.  First,  the option "Fl-Enter name of data file used during
execution" is used, and  the filename HOPPER is entered after selecting  this option.    Once the
filename is entered, the options "F4-Print selected output file" or "F5-View selected output file" can
be used.  There is considerable output from the model and all of the output  is usually not desired.
The F5 option provides viewing of the output file by paging through it using the PAGE UP and
PAGE DOWN keys.  The viewing software can be used to select specific portions of the output for
printing a hard copy.  Pressing the ESC key returns the "STFATE Activity Selection Menu".
                                                                                DRAFT

-------
                                             C-53

C2.8.2.2.1        Hopper Disposal Water Column Concentrations and Area Distribution

In this  example, water column concentrations  of the  solids fractions  and the fluid volume ratio
(volume of dumped fluid/volume of  ambient water) are requested  at 20 and 39 ft.   Thus,  the
concentrations for sand, silt,  clay  and the fluid  at every grid point location  for both  depths  are
contained in the output file provided the material has not settled to the bottom.  In addition, results
are also available showing the maximum concentration occurring at each grid point for anywhere in
the water column as well  as at the  requested depths throughout the duration of the simulation.  The
top portion of the selected output  shown  in  Figure C-14 shows  the output for fluid volume ratio
(volume of fluid from the  discharge / volume of water column at the grid point)  at 20 ft at the end of
the model run (3600 s). These values at each  grid point are multiplied by the appropriate scale factor
(0.01) as given at the top of the  output. These values would  be multiplied by 100 if the results were
desired  with units of  percent.   In this example the maximum value on the  grid is 1.6 which is
multiplied by 0.01 yielding a maximum fluid volume ratio of 0.016 at  x- and z-grid locations of 21
and  16  respectively.   Recall from  the input  that the distance between  x-grid points  is  150 ft  and
between z-grid points is 50 ft. Therefore the maximum concentration occurs 3000 ft from top of grid
and 750 ft from left edge of grid. The disposal began at an x- and  z-distance of 1000 ft (grid  point 8)
and 750 ft (grid point 16).  Both the hopper dredge and water velocity were in the positive x-direction
so it is  reasonable to find the maximum concentration down grid from the disposal point.  Since the
hopper  and the water current  are moving in the positive x-direction, it is expected that the distance in
the x-direction affected by the discharge should be longer than that in the z-direction.  The display of
the area with  concentrations greater than  10~8 vol/vol in Figure C-14 appears to be wider than  it is
long, but that is because of the difference in grid spacing  between the x- and z-direction.  It is
actually 800 ft wide (z-direction) and 1500 ft long (x-direction).
C2.8.2.2.2        Hopper Water Column Concentrations

The  water  column concentrations over the duration of the simulation  are tabulated  in Figure C-14.
This shows the clay and percent fluid values versus time.  The sand settled to the bottom before 900 s
so there  is no history for this  sediment fraction.  Maximum concentrations for silt  and clay  on the
entire grid  are shown to decrease with time for both requested depths  and the quarterly time entered
in the input data while outside  the mixing zone the concentrations increased as the plume moved out
from the mixing zone.   The fluid volume ratio is shown for every time step and at the two depths of
20 and 39 ft as well as the model selected depths of 0, 10, 20, 30, and 40 ft.  The simulation
                                                                               DRAFT

-------
                                                         C-54
CONCENTRATIONS ABOVE BACKGROUND OF FLUID (VOLUMETRIC RATIO OF DUMP FLUID TO AMBIENT WATER) M THE CLOUD 3600 00 SECONDS AFTER DUMP

       20.00 FT BELOW THE WATER SURFACE
 ...MULTIPLY DISPLAYED VALUES BY 0.1000E-01  (LEGEND  + . .LT. .01 . - .LT  0001 0 > .LT. .000001)
 MN- 2  3   4   6   6   7   8   9   10   11  12  13   14  IE  16  17   18  18  2O  21  22  23   24  26 26 27  28  28 30  31
3OOOO
40000
6 QOOO
6OOOO
70OOO
8 OOOO
eoooo
to oooo
11 OOOO
12 OOOO
130000
14 OOOO
IS OOOO
18 OOOO
17 OOOO

18 OOOO
200000
21 OOOO
22 OOOO
230000
24 OOOO
26 OOOO
28 OOOO
270000
28 OOOO
28 OOOO
30 OOOO
0
0
Q
0
0
0
0
0
0
0
0
0
0
Q

0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
A

0
0
0
0
0
0
0
0
0
0
0
0
0
0
Q
0
0
0
0
0
0
0
0
0
0
0
Q

0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
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A

0
0
0
0
0
0
0
0
0
0
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0
0
0
0
0
0
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0
0







0
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0

+
01
02
.02
.01
+
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0

+
03
07
06
.02
+
0
0
0
0
0
0
0
Q
0
0
Q
0
0
0
0
0
0
0
0

.01
06
16
16
.06
.01
0
0
0
0
0
0
0
Q
0
0
0
0
0
0
0
0
0
0

03
.16
.33
.30
.13
.02
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0


.06
.28
.68
.63
.23
.04
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0


.08
.48
BO
83
36
.06
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0


.13
63
1.2
1.1
.48
.08
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0


.01
.18
.76
1.6
1.3
.68
.10
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0

*
.01
.17
.81
1.6
1.4
62
.10
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0

+
.01
.16
.76
1.6
1.3
.68
.10
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0


.01
.13
.83
1.2
1.1
.48
.08
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0


.08
.46
.80
.83
.36
06
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0


.06
.28
.68
.63
.23
.04
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0

.03
.16
.33
.30
.13
.02
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0

.01
.08
.16
.16
.06
.01
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0

0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
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0
0
0
0
0
0
0
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0
0
0
0
0
0
0
0
0
0
0
0
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0
0
0
0
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0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
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0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
A

0
0
0
0
0
0
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0
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0
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0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
A

0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
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0
0
0
A

0
0
0
0
0
0
0
0
0
0
0
OOOO
OOOO
OOOO
OOOO
OOOO
OOOO
OOOO
OOOO
OOOO
OOOO
OOOO
OOOO
OOOO

OOOO
OOOO
OOOO
OOOO
OOOO
OOOO
OOOO
OOOO
OOOO
OOOO
OOOO
OOOO
 31 OOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOO
INITIAL NIXING COMPUTATIONS RESULTS FOR  SILT:


TINE
(HR)
0.25
0.50
0.75
1.00
0.25
0.50
0.75
1.00
INITIAL NIXING


TINE
(HR)
0.25
0.50
0.75
1.00
0.25
0.50
0.75
1.00


DEPTH
(FT)
20.0
20.0
20.0
20.0
39.0
39.0
39.0
39.0
COMPUTATIONS


DEPTH
(FT)
20.0
20.0
20.0
20.0
39.0
39.0
39.0
39.0
MAX CONC ABOVE
BACKGROUND
ON ENTIRE GRID
(MG/L)
0.493E+03
0.456E+03
0.311E+03
0.217E+03
0.137E+04
0.751E+02
0.511E+02
0.357E+02
RESULTS FOR CLAY
MAX CONC ABOVE
BACKGROUND
ON ENTIRE GRID
(MG/L)
0.477E+03
0.299E+03
0.209E+03
0.147E+03
0.244E+03
0.492E+02
0.343E+02
0.241E+02


X-LOC
(FT)
1800.
2100.
2550.
3000.
1650.
2100.
2550.
3000.
:


X-LOC
(FT)
1800.
2100.
2550.
3000.
1500.
2100.
2550.
3000.


Z-LOC
(FT)
750.
750.
750.
750.
750.
750.
750.
750.



Z-LOC
(FT)
750.
750.
750.
750.
750.
750.
750.
750.
MAX CONC ABOVE
BACKGROUND OUTSIDE
MIXING ZONE
(MG/L)
0.504E-04
0.146E-01
0.507E+00
0.217E+03
0.162E-04
0.240E-02
0.834E-01
0.357E+02

MAX CONC ABOVE
BACKGROUND OUTSIDE
MIXING ZONE
(MG/L)
0.476E-04
0.215E-01
0.516E+00
0.147E+03
0.214E-04
0.353E-02
0.849E-01
0.241E+02
Figure C-14.   Selected Output for Hopper Dredge Disposal.
                                                                                                      DRAFT

-------
                                           C-55
INITIAL NIXING COMPUTATIONS RESULTS FOR FLUID:
MAX CONC ABOVE MAX CONC ABOVE
BACKGROUND BACKGROUND OUTSIDE
TIME DEPTH ON ENTIRE GRID X-LOC Z-LOC
(HR) (FT)
0.08 20.0
0.17 20.0
0.25 20.0
0.33 20.0
0.42 20.0
0.50 20.0
0.58 20.0
0.67 20.0
0.75 20.0
0.83 20.0
(PERCENT) (FT) (FT)
0.203E+01 1350. 750.
0.548E+01 1500. 750.
0.485E+01 1650. 750.
0.427E+01 1800. 750.
0.376E+01 1950. 750.
0.331E+01 2100. 750.
0.292E+01 2250. 750.
0.258E+01 2400. 750.
0.228E+01 2550. 750.
0.202E+01 2700. 750.
THE TOXICITY STANDARD IS VIOLATED OUTSIDE THE MIXING ZONE AT
0.92 20.0
0.180E+01 2850. 750.
THE TOXICITY STANDARD IS VIOLATED OUTSIDE THE MIXING ZONE AT
1.00 20.0
0.160E+01 3000. 750.
THE TOXICITY STANDARD IS VIOLATED OUTSIDE THE MIXING ZONE AT
0.08 39.0
0.17 39.0
0.25 39.0
0.33 39.0
0.42 39.0
0.50 39.0
0.58 39.0
0.67 39.0
0.75 39.0
0.83 39.0
0.92 39.0
1.00 39.0
0.08 0.0
0.17 0.0
0.25 0.0
0.33 0.0
0.42 0.0
0.50 0.0
0.58 0.0
0.67 0.0
0.75 0.0
0.83 0.0
0.92 0.0
0.245E+01 1500. 750.
0.902E+00 1500. 750.
0.797E+00 1650. 750.
0.702E+00 1800. 750.
0.618E+00 1950. 750.
0.544E+00 2100. 750.
0.480E+00 2250. 750.
0.424E+00 2400. 750.
0.375E+00 2550. 750.
0.332E+00 2700. 750.
0.295E+00 2850. 750.
0.263E+00 3000. 750.
0.135E+01 1350. 750.
0.148E+01 1500. 750.
0.131E+01 1650. 750.
0.116E+01 1800. 750.
0.102E+01 1950. 750.
0.896E+00 2100. 750.
0.789E+00 2250. 750.
0.697E+00 2400. 750.
0.617E+00 2550. 750.
0.547E+00 2700. 750.
0.486E+00 2850. 750.
THE TOXICITY STANDARD IS VIOLATED OUTSIDE THE MIXING ZONE AT
1.00 0.0
THE TOXICITY STANDARD
0.08 10.0
0.17 10.0
0.25 10.0
0.33 10.0
0.42 10.0
0.50 10.0
0.58 10.0
0.67 10.0
0.75 10.0
0.83 10.0
0.92 10.0
THE TOXICITY STANDARD
1.00 10.0
THE TOXICITY STANDARD
0.433E+00 3000. 750.
IS VIOLATED OUTSIDE THE MIXING ZONE AT
0.672E+01 1350. 750.
0.338E+01 1500. 750.
0.299E+01 1650. 750.
0.264E+01 1800. 750.
0.232E+01 1950. 750.
0.204E+01 2100. 750.
0.180E+01 2250. 750.
0.159E+01 2400. 750.
0.141E+01 2550. 750.
0.125E+01 2700. 750.
0.111E+01 2850. 750.
IS VIOLATED OUTSIDE THE MIXING ZONE AT
0.989E+00 3000. 750.
IS VIOLATED OUTSIDE THE MIXING ZONE AT
MIXING ZONE
(PERCENT)
0.823E-26
0.305E-05
0.559E-04
0.480E-03
0.240E-02
0.794E-02
0.195E-01
0.386E-01
0.649E-01
0.627E+00
0.83 HOURS.
0.162E+01
0.92 HOURS.
0.160E+01
1.00 HOURS.
0.805E-26
0.503E-06
0.920E-05
0.789E-04
0.395E-03
0.131E-02
0.321E-02
0.634E-02
0.107E-01
0.103E+00
0.267E+00
0.263E+00
0.560E-26
0.735E-06
0.151E-04
0.130E-03
0.649E-03
0.215E-02
0.528E-02
0.104E-01
0.176E-01
0.170E+00
0.439E+00
0.92 HOURS.
0.433E+00
1.00 HOURS.
0.251E-25
0.180E-05
0.345E-04
0.296E-03
0.148E-02
0.490E-02
0.120E-01
0.238E-01
0.401E-01
0.387E+00
0.100E+01
0.92 HOURS.
0.989E+00
1.00 HOURS.
Figure C-14.  Selected Output for Hopper Dredge Disposal, (continued)
                                                                           DRAFT

-------
                                                  C-56
0.08
0.17
0.25
0.33
0.42
0.50
0.58
0.67
0.75
0.83
20.0 0.122E+02 1500. 750.
20.0 0.548E+01 1500. 750.
20.0 0.485E+01 1650. 750.
20.0 0.427E+01 1800. 750.
20.0 0.376E+01 1950. 750.
20.0 0.331E+01 2100. 750.
20.0 0.292E+01 2250. 750.
20.0 0.258E+01 2400. 750.
20.0 0.228E+01 2550. 750.
20.0 0.202E+01 2700. 750.
THE TOX1C1TY STANDARD IS VIOLATED OUTSIDE THE MIXING ZONE AT
0.92
20.0 0.180E+01 2850. 750.
THE TOXICITY STANDARD IS VIOLATED OUTSIDE THE NIXING ZONE AT
1.00
20.0 0.160E+01 3000. 750.
THE TOXICITY STANDARD IS VIOLATED OUTSIDE THE MIXING ZONE AT
0.414E-25
0.305E-05
0.559E-04
0.480E-03
0.240E-02
0.794E-02
0.195E-01
0.386E-01
0.649E-01
0.627E+00
0.83 HOURS.
0.162E+01
0.92 HOURS.
0.160E+01
1.00 HOURS.
0.08
0.17
0.25
0.33
0.42
0.50
0.58
0.67
0.75
0.83
0.92
30.0 0.798E+01 1500. 750.
30.0 0.333E+01 1500. 750.
30.0 0.294E+01 1650. 750.
30.0 0.259E+01 1800. 750.
30.0 0.228E+01 1950. 750.
30.0 0.201E+01 2100. 750.
30.0 0.177E+01 2250. 750.
30.0 0.156E+01 2400. 750.
30.0 0.138E+01 2550. 750.
30.0 0.123E+01 2700. 750.
30.0 0.109E+01 2850. 750.
THE TOXICITY STANDARD IS VIOLATED OUTSIDE THE MIXING ZONE AT
1.00
30.0 0.971E+00 3000. 750.
THE TOXICITY STANDARD IS VIOLATED OUTSIDE THE MIXING ZONE AT
0.251E-25
0.187E-05
0.339E-04
0.291E-03
0.146E-02
0.481E-02
0.118E-01
0.234E-01
0.393E-01
0.380E+00
0.984E+00
0.92 HOURS
0.971E+00
1.00 HOURS
          0.08
          0.17
          0.25
          0.33
          0.42
          0.50
          0.58
          0.67
          0.75
          0.83
          0.92
          1.00
40.0
40.0
40.0
40.0
40.0
40.0
40.0
40.0
40.0
40.0
40.0
40.0
0.167E+01
0.742E+00
0.656E+00
0.578E+00
0.509E+00
0.448E+00
0.395E+00
0.348E+00
0.308E+00
0.273E+00
0.243E+00
0.217E+00
1500.
1500.
1650.
1800.
1950.
2100.
2250.
2400.
2550.
2700.
2850.
3000.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
750.
0.560E-26
0.412E-06
0.757E-05
0.649E-04
0.325E-03
0.107E-02
0.264E-02
0.522E-02
0.878E-02
0.848E-01
0.220E+00
0.217E+00
 RESULT:   THE TOXICITY  STANDARD WAS VIOLATED OUTSIDE  THE MIXING ZONE DURING THE SIMULATION.

Figure C-14.  Selected Output for Hopper Dredge Disposal, (concluded)
                                                                                         DRAFT

-------
                                             C-57
indicates the 0.4% toxicity standard entered in the input phase is violated at depths of 0, 10, 20, and
30 ft, but it is not violated at 39 and 40 ft within the 3600 seconds. Thus the statement at the bottom
of Figure C-14 says the toxicity standard is violated outside the mixing zone.
C2.8.2.2 J     Hopper Maximum Concentration and Contour Graphs

From the "STFATE  Activity Selection Menu," "F4-Generate  Graphics"  is selected  to  receive the
"STFATE Graphics File Selection Menu."  First, the name of the data file used during execution is
entered  after selecting "Fl-Enter name of data file used during execution."  Next, the "F4-Generate
graphics with selected file" is pressed to receive the "STFATE Graphics Generation Menu."  The first
option is to  select  "Fl-Maximum concentrations versus time" which brings  a  screen up  to select
where and what to plot.  Either the screen, printer or plotter must be selected and then the material
(sand, silt,  clay or fluid) and depth (20, 39 or peak).  Peak means the depth at which the maximum
concentrations or fluid ratio  occur.  The maximum  fluid ratio versus time in this hopper disposal
example (Fig.  C-15)  shows  the maximum percent (12.2%) on the grid  occurs about  5 min after
disposal begins and it drops to just below 1.6% near the end of the simulation.  The maximum fluid
percent  outside the mixing zone, defined in the input, is initially below the toxicity standard (0.4%),
but the  standard is violated at  about 49 min after disposal begins  and  remains  in violation  for the
remainder of the simulation.  Referring back to Figure C-14, it can  be seen that the peak depth is 20
ft.
          C
          0
          N
          C
          v
          0
          1
14

12

10

 8

 6

 A

 2

 9
                     HOPPER DISCHMO UITH SPEC IF I ED MIXING ZONE (TIER  III)
                                                                             FLJD
                                                                                  FT
                                      24     32     48
                                       TIME - Hi mi tea
                                                  56
                                     ilDE M.Z.
                                 —  M.Z. STANDARD
Figure C-15.   Peak Fluid Ratio as a F(Time) for the Hopper Dredge Disposal Example.
                                                                               DRAFT

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                                             C-58
Selecting option "F2-Concentration contours in horizontal plane" displays a screen which provides the
ability to graphically display the percent fluid contours or the solids fraction concentration contours.
As before, the graphs can be output to the screen, plotter or printer.  The contours are obtained by
selecting the solid fraction, fluid or mixing zone and then selecting the depth desired (20, 39, or peak
in this example).  Default contours are selected by answering  "YES" or user-specified contours are
selected by answering "NO".  Figure C-16 shows one fluid percent contour at 20 ft for 3600 s after
initiation of disposal.  The  contour value was specified for only one  contour  and the toxicity value
(0.4%) was entered. Thus, Figure C-16 shows the standard was exceeded just outside the predefined
mixing zone at that time. Figure  C-17 shows the required mixing  zone boundary up to 3600 s after
disposal as outlined by  the 0.4% contour.  Since  this contour falls outside the specified mixing, the
figure shows a violation of the standard.  The  highlighted box with  "Mixing" is selected  from the
screen appearing after pressing the F2 option on the "STFATE Graphics Generation Menu"  to obtain
this result.  The ESC key is now pressed repeatedly to return to the "STFATE Activity  Selection
Menu."
                 HOPPER DISCHARGE UITH SPECIFIED NIXING ZONE (HER III)
                                  0
            0
            N
                                1268
                                24B8
                                3680
                                                            FLUID
                                                             PERCENT
                                                           ZB.BB  FT
                                                           %ae. SEC
                                                       COHTOUS UALUES
                                                       1 "  4.0BBE-B1
                                     8           1606
                                      Z DIRECTION
Figure C-16.
Fluid Ratio Contours for 20 ft Depth at 3600 sec for the Hopper Dredge Disposal
Example.
                                                                                DRAFT

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                                             C-59
                 HOPPER DISCHARGE UIIH SPECIFIED NIXING ZONE  (HER III)
                                  B
            T
            I
            0
                               1288
                               Z4B8
                               36B8
     NIXING
      PERCENT
   PEAK   PT
    X0B. SEC
CONTOUR VALUES

1 M  4.BBBE-01
                                                1680
                                      Z DIRECTION
Figure C-17.    Plot of Required Mixing Zone for the Hopper Disposal Example.
                                                                                DRAFT

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                                             C-60

C3.0      CORNELL MIXING ZONE EXPERT SYSTEM (CORMIX)

The Cornell Mixing  Zone  Expert System  (CORMIX) is a steady state  three-dimensional  model
(Donekar  and Jirka, 1990).   CORMIX was developed to predict the dilution and  trajectory of  a
submerged single port discharge of arbitrary density  (positive, neutral, or negative) into a stratified or
uniform-density ambient environment with or without cross-flow.  CORMIX is an integral model that
accounts for most  near-field  and some  far-field  steady  state dynamics.   CORMIX is presently
designed for use in shallow  water systems where the jet mixing processes are expected to encounter
bottom  boundary  interaction.   CORMIX is  capable of representing  negatively buoyant  plume
dynamics through application of mirroring principals;  however, the present version does not include
sediment settling and deposition.

The current version of the CORMIX model  requires some modifications to extend its capabilities to
simulate the characteristics of dredged  material discharges.  Efforts  are underway for adaptations of
the CORMIX model for  simulating the mixing hydrodynamics of several  types of dredged material
discharges.  When these efforts  are completed, the  revised CORMIX model  will  be included in
subsequent revisions of this appendix1.
       The latest release of CORMIX (Version 2.10) can be obtained without charge from U.S. EPA Office of
       Research and Development, Center for Exposure Assessment Modeling (CEAM), Athens Environmental
       Research Laboratory, 960 College Station Road, Athens, Georgia  30605-2720. CORMIX can be either
       downloaded from CEAM's on-line Bulletin Board System by calling 1-706-546-3402 (FTS 250 3402),
       or sent through the mail by sending user-supplied diskettes or 9-track magnetic tapes to the CEAM
       Model Distribution Coordinator at the above address. User documentation is also available from the
       same source.
                                                                               DRAFT

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                                            C-61
C4.0     MACINTYRE  ANALYTICAL  METHOD  FOR  CDF DISCHARGE  IN  RIVERINE
          CONDITIONS

C4.1     Introduction

This section presents a simplified approach that is applicable to relatively shallow confined riverine
water bodies.   The method involves a simplistic  two-dimensional calculation based on dispersion
principles (Maclntyre, 1987).  If the mixing-zone size as calculated using simple approximations is
within mixing-zone guidelines specified by regulatory agencies, more precise calculations may not be
necessary.  The mixing-zone calculations depend on a number of assumptions that are difficult to
satisfy for estuaries and the tidally influenced portions of rivers.  The difficulties are discussed after
the presentation of the procedure to be used for a riverine environment.

The analytical  solution technique for calculating mixing-zone size described in this section is  based on
theoretical and empirical relationships for dispersion as summarized by Fischer et al.  (1979).  Only
equations for calculating mixing-zone size resulting from a single-point discharge are presented.

A schematic illustrating a typical single-source  effluent  discharging into a receiving  water body is
shown in Figure C-18.  For such a condition, the mixing-zone length extends downstream and  the
body of the mixing zone remains close to the shoreline of the receiving water body.
                                   RECEIVING WATER


                                       OUTFALL
                                   DREDGED
                                   MATERIAL    WEIR
                                 CONTAINMENT
                                     AREA
Figure C-18.    Schematic of a Mixing Zone for a Single Effluent Source.
                                                                             DRAFT

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                                              C-62

C4.2      Data Requirements

The following data are required for evaluating mixing-zone sizes for confined disposal area effluents:

       a. Effluent concentrations  at  the  point  of  discharge and  receiving  water  background
          concentrations for all contaminants of concern.
       b. Water  quality  standards applicable  at  the  limit  of the allowable  mixing  zone for  all
          contaminants of concern.
       c. Depth,  cross-sectional  area,  and current velocity  of the receiving water  body  during
          expected low flow conditions during the period of dredging.
       d. Effluent volumetric flow rate.


C4.3      Calculation Procedure

Step 1.   Verify that the assumptions on which the equations depend are reasonable for conditions at
          the proposed discharge  site.

Step 2.   Use  effluent, receiving  water, and water quality standard concentrations of all contaminants
          of concern to identify  the critical contaminant.   The critical contaminant  is the one that
          requires the greatest dilution,  which will define  the boundary of the mixing zone.   If
          mixing  evaluations are conducted for toxicity test results the background  concentration of
          dredged material is assumed to be zero and the percentages of dredged material are used to
          calculate the required dilution.

Step 3.   Use  receiving-water depth and velocity data to calculate a lateral mixing coefficient. This
          coefficient is a measure of how rapidly the effluent is dispersed through the receiving
          water.

Step 4.   Calculate mixing-zone length.

Step 5.   Check assumptions that depend on mixing-zone length.

Step 6.   Calculate the maximum width of the mixing zone.

Step 1 - Assumptions.  In order to apply the analytical solution described in this section, the following
assumptions are required:
                                                                                  DRAFT

-------
                                              C-63

       a. No major change  in  cross-sectional shape,  sharp  bends, major  inflows or outflows, or
          obstructions to flow exist in the receiving water body in proximity to the mixing zone.
       b. The receiving water body can be reasonably approximated by a shallow rectangular cross
          section.
       c. The confined disposal  area effluent enters  the receiving water as a point source at the bank
          with negligible horizontal momentum.
       d. Differences in density between the  effluent  and receiving  water and  in settling  rates of
          suspended particles within the boundary of the mixing zone are negligible.
       e. The flow condition in  the vicinity of the mixing zone can be  approximated as a steady-state
          velocity flowing parallel to the bank of the receiving water.
       f. The major cause of dispersion in the receiving water body is the turbulence and shear flow
          associated with the horizontal water flow.
       g. The effluent plume is vertically well  mixed, so  that contaminant concentrations do not vary
          significantly with depth.
       h. The width of the effluent plume is small enough  that its lateral dispersion is not restricted
          by the opposite bank of the receiving water body.

Step 2 - Identify critical contaminant.  It is necessary to calculate the dilution  required within the
mixing zone in order to reach  applicable water quality  standards  for all  contaminants of concern.
This requires an estimate of the effluent concentrations of regulated contaminants.  The contaminant
that requires the greatest amount  of dilution should be calculated as described in Section 5.3.

The  maximum  boundary  of the  mixing zone  will be  defined as the  isopleth  (line of  constant
concentration) where the concentration of the most critical contaminant is reduced  to the concentration
specified  by  the appropriate water quality  standard.    It  should be noted  that if background
concentrations exceed the water quality standard, the  concept of a mixing zone is inapplicable.

Also, this approach for calculating required  dilution  is not applicable to turbidity  (an optical property
of water), which is  reduced in an nonlinear fashion  by dilution.  A correlation curve for TSS versus
turbidity should be used to define the TSS concentration  corresponding to the water quality standard
for turbidity.

Step 3 - Estimate of lateral mixing coefficient.
Step 3.1. The depth of a simplified rectangular cross section for the receiving water body should be
calculated as follows:
                                                                                  DRAFT

-------
                                              C-64
where
       d =  average depth of the receiving water body channel, m
       A = cross-sectional area of the channel, m2
       W = surface width of the channel, m
Check to ensure that  W   is equal to or greater than 10 times the average depth d.   If not, the
estimate of a lateral mixing coefficient is likely to be inadequate.

Step 3.2.  Estimate the shear velocity by one of the following methods.  In rivers where the mean
channel slope is known, use:

                                          u*  = \lgds
In rivers where the channel slope is not known, use:
                                          u* = o.i u
where
       u* = shear velocity in receiving water, m/sec"1
       g =  gravitational acceleration, 9.81 m/sec"2
       d =  average channel depth, m
       S =  slope of river bed (dimensionless)
       u =  average of instantaneous velocities across the channel cross section, m/sec"1.

If the flow rate of the receiving water is known, u can be calculated as the flow rate divided by the
channel cross-sectional  area.  If the receiving-water flow rate is not known, u must be determined
from  velocity measurements taken at the proposed  site.  It should be noted that u should not be
determined  over a  period  of time during  which  velocity changes  occur  due to changes in the
receiving-water flow rate.

Step 3.3.  Estimate the lateral mixing coefficient by using one of the following equations.
In rivers:
                                                                                 DRAFT

-------
                                             C-65
                                         Et = 0.3 cfu*
In estuaries:
                                         E. - 0.4 du*
where
       E, = lateral mixing coefficient, nWsec"1
       d = average channel depth, m
       u* =  shear velocity, m/sec"1

The values of  lateral mixing coefficient are derived from Fischer et al.  (1979)  and are based on
experimental  studies  of dispersion in various rivers.  Lateral mixing coefficients have been shown to
vary widely from one location to another, and the above equations give the lowest reasonable values
so that estimates of mixing zone size will be conservative.

Step 4 - Estimate mixing-zone length. If the assumptions presented earlier are valid,  the mixing zone
will have a shape similar to the one shown in Figure C-18. The length of the mixing zone (measured
parallel to the bank) can be estimated as:
                                   L  *[**fiij[
-------
                                             C-66

where
       L = predicted mixing zone length, m
       u = cross-sectional average velocity (instantaneous or averaged over a few minutes), m/sec'1
       Tc = time taken for the observed value of u to change by 10 percent, in seconds

Step 5.2.   The lateral dispersion of the effluent plume will not be restricted by  opposite bank of the
receiving water body as long as:
where W = surface width of receiving water channel, m.

Step  6  -  Estimate maximum width  of mixing  zone.   The  maximum width  of the mixing zone
(measured perpendicular to the bank as shown in Figure C-18 can be estimated as:


                                        v _  0.4840Q,C,
                                            u(C, - Cb)d
where Y = maximum width of the mixing zone, m.
C4.4     Example Mixing-Zone Calculation

Following is a hypothetical mixing-zone calculation designed to illustrate the use of the mixing-zone
estimation equations.  A proposed dredged material containment area is expected to discharge into a
river 480 ft  (146.3 m) wide.  From a study of US Geological Survey stream gage  records, it is
anticipated that while  effluent will be discharged, the lowest river  flow will be about 7,600 rVVsec
(212.8 nrYsec) and that the river has a cross-sectional area of 4,000 ft2 (371.6 m2) at this flow rate.
The local  bed slope of the river is known to be very variable due  to sediment transport.  The
containment  area is expected to  have a peak discharge of 15  cfs. The only effluent contaminant that
exceeds water quality standards  will  be cadmium, which  is expected  to have an effluent concentration
of 3.5 ngfL.  The background concentration of cadmium in  the river is below the detection limit of
0.1 jtg/L, and the applicable cadmium water quality standard is 0.25 /ig/L.  It has been specified that
the  maximum  acceptable  mixing-zone  size  is  a 750-ft  (228.6-m)  radius  centered  on the effluent
outfall.
                                                                                DRAFT

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                                             C-67

Step 1 - Assumptions.  Since the purpose of this hypothetical problem is to demonstrate the use of the
mixing-zone calculations,  it has been defined so  that all the assumptions on which the calculations
depend are valid.  Decisions on whether the assumptions are valid depend largely on the professional
judgement of personnel familiar with the disposal site.

Step 2 - Identify critical contaminant.  Cadmium  is the only effluent contaminant that exceeds water
quality standards for this example.  It is therefore unnecessary to determine the critical contaminant.

Step 3 - Estimate lateral mixing coefficient.

Step 3.1.  From the problem statements,

                                  A = 4,000 ft2 (371.6 m2)
                                    V = 480 ft (146.3 m)
Calculate depth from equation 2:
                                                 A
                                                 T7
                                   *-  3,7A'6'a =2.54m
                                        146.3 m
Check that W £ 10 d .  It is.

Step 3.2.   Since the local bed slope is known to vary due to sediment transport, the shear velocity
should be estimated from the mean velocity.  Calculate the mean velocity by dividing the river flow
of 7,600 fVVsec (212.8 nvYsec) by the cross-sectional area of 4,000 ft2 (371.6 m2):
                     - = 7,600 cfs = 1>9£) ftfBec-L  (0.579 m/sec'l)
                          4,000 ft2
                                                                               DRAFT

-------
                                           C-68

and calculate the shear velocity of the receiving waters as follows:


                                        u* « o.i u


                        u*  =  0.1(0.579 m/secr1) = 0.0579  ra/sec'1
Step 3.3. In rivers, the lateral mixing coefficient should be estimated as:
                                       Et = 0.3 d u*
                            E, = 0.3(2.54 m) (0.0579 m/secr1)
                                   E, = 0.0441 fli2/sec
Step 4 - Estimate mixing-zone length.  Estimate using the problem statements:

                               Qe = 15 cfs (0.425 m3/
                             Ct = 3.5 pg/L-'  (3.5 x
                            C, - 0.25 fjg/L'1  (2.5 x 10"4 mg/L'1)
                            Ch< 0.1 j/g/L-1 (1.0 x  10-4 mg/L'1)
In order to be conservative, it would be wise to assume that the background concentration is only just
under the detection limit, rather than zero. Therefore use:
                                                                             DRAFT

-------
                                           C-69
                                  Cb - 1.0 x 10"4 mg/L'
Calculate mixing-zone length:

                              ir(0.0441 mVsec-1) (0.579 m/secr1)
                            (0.425 m2/sec) (3.5 x 10'3i
                               .5 - 1.0) x losing L~'] (2.54 m).
                                   L = 190 m (623 ft)
Step 5 - Check length-dependent assumptions.
Step 5.1:

Verify that the flow of the water body near the mixing zone can be treated as a steady state flow.
therefore:
                                                                           DRAFT

-------
                                          C-70
                                      >  10(190 «)
                                     c   0.579 in/sec-1
                                Tc 2 3,280 sec  (55 min)



This is acceptable since the river flow will certainly not change by 10 percent in less than 1 hour.

Step 5.2:
                                  8(0.0441 jnVsec'1) (190 m)
                                        (0.579
                                       W > 10.8 m



This condition is amply satisfied since W equals 146 m.


Step 6 - Estimate maximum width of mixing zone.  Estimate the maximum mixing zone width as:


                                     r _ 0.484 Q.C,
                                         U(C, - Cb)d
                  Y =     0.484  (0.425 m3/sec-') (3. 5 x 1Q'3 mg/L"')
                      0.579 m/sec-'l (2.5 - 1.0) x 1Q-" mgr/L'1] (2.54 m)
                                   Y = 3.3 m (10.7 ft)
                                                                           DRAFT

-------
                                            C-71

Since the mixing zone is predicted to have a length of 623 ft (190 m) and a maximum width of 10.7
ft (3.3 m), it is within the allowable limits of 750 ft (228.6 m) from the effluent outfall.
                                                                             DRAFT

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                                            C-72

C5.0     FASTTABS MODELING  SYSTEM  FOR  EVALUATION  OF  HYDRODYNAMIC
         TRANSPORT

Rivers, reservoirs, and estuaries have been modeled for a number of years using the USAGE TABS
numerical modeling  system.   TABS  is  a family of two-dimensional numerical models  that  can
simulate hydrodynamic, sediment, and constituent transport processes in these water bodies.  TABS
has been used to simulate far-field dispersion of instantaneous and continuous  dredged material
discharges.  Some independent near-field analysis is usually required.  TABS can handle complex
geometries  and unsteady flow conditions.  Either paniculate or dissolved phases of dredged material
can be modeled.

The TABS system consists of many separate programs that individually address different aspects of
the modeling process (Thomas and  McAnally, 1990).  These include mesh development, geometry
input  file generation, boundary condition definition,  hydrodynamic input  file generation, job  status
monitoring, and post-processing of the results.

A new graphical  implementation  of TABS (FastTABS)  (Lin et al., 1991) has been developed that
successfully addresses the need for efficient model setup, execution,  and analysis.  It is mouse driven
with pull down menus and requires a minimum of manual data entry to complete an application from
start to finish.  FastTABS  was designed to allow easy application of each of the models in the TABS
system which  include hydrodynamics, constituent and sediment transport.  The FastTABS software
runs on  Macintosh and DOS-based personal computers  as well as  most UNIX  workstations.   A
primer, user's manual, and tutorial are available2.
   2 A limited government license allows USAGE office use of the FastTABS software supplied through the USAGE
Waterways Experiment Station (WES). Other users may obtain the software from Brigham Young University, (801)-378-
5713.  The point of contact for additional information is: Mr. David R. Richards, USAGE Waterways Experiment Station,
ATTN: CEWES-HE-S, 3909 Halls Ferry Road, Vicksburg, MS 39180-6199, (601)-634-2126.
                                                                              DRAFT

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                                            C-73

C6.0     DILUTION VOLUME METHOD FOR CDF EFFLUENT DISCHARGES

C6.1     Approach

A simplified approach to evaluation of mixing zones for CDF effluent discharges is presented in this
section  in  which  the volume of  water required  for dilution  is  expressed  as a  rate  of  flow
(Environmental Effects Laboratory,  1976). This approach is generally applicable in both riverine and
estuarine conditions.  However,  the approach should  only  be applied  where there is  a discrete
discharge source such as a conduit or a weir.  Since the effluent discharge will occur at a specified
rate Vp, the volume of ambient site water per unit time that would be required to dilute the discharge
to acceptable levels can be defined as:
                                           ((c* -
where
       VA = volume of site water/unit time required for dilution, cfs
       Vp = rate of effluent discharge, cfs
       Ce = concentration of the contaminant in the effluent in /ig/L
       CBQ = background concentration of the contaminant in the disposal site
          water in jug/L
       CWQ  = applicable water quality standard for the contaminant in /xg/L

It is assumed that the mixing zone associated with an effluent discharge will  resemble the shape in
Figure C-19. Therefore, once the required volume per unit time has been calculated, the next step is
to  determine the  dimensions of the mixing zone.  The  required volume per  unit time can also be
expressed as:
                                         v, = L d va
where
       VA = required volume of water per unit time, cfs
       L =  width of mixing zone at time t, ft
       d = depth, ft
       Vw = velocity of water at disposal site, ft/sec
                                                                              DRAFT

-------
                                            C-74
                    PLAN
        PROJECTED SURFACE AREA
 FRONTAL
ELEVATION
                                                                  VOLUME PER UNIT TIME
               A=  |  — + r I  X
                      2
   VA=LdVw
Figure C-19.    Projected  Surface Area and Volume Equations for CDF Effluent Discharge with
               Prevailing Current.

Since the depth and water velocity are known or can be measured, the width of the front edge of the
mixing zone can be calculated as:
                                          L =
                                              d V.
Based on Brooks (1960) and Johnson and Boyd (1975), the time required for the front edge of the
mixing zone to spread laterally to the required width L can be computed from:
                                                                             DRAFT

-------
                                              C-75
                                t = -i  (0.094 L2/3 - 0.149 r2/3)
where
       t = required time for lateral spreading, sec
       L = necessary width of the front edge of mixing zone, ft
       r = one-half initial width of the plume at point of discharge (radius of initial surface mixing),
          ft
       X = turbulent dissipation parameter

Values for  X  range from 0.00015 to 0.005 with a value of 0.005 being appropriate in a dynamic
environment such as  an estuary (Brandsma and Divoky, 1976).   As discussed earlier, values for  r
will be influenced by the method of disposal and will be site specific.

The calculated time can then be used to determine the longitudinal distance the discharge will travel as
it is spreading to the required width.  This distance can be computed from:
where
       X = longitudinal movement of discharge, ft
       Vw = velocity of water at disposal site, ft/sec
       t = necessary time of travel, sec

The results of the previous  equations can then be combined to estimate the projected surface area of
the proposed discharge.  This  area can be computed as:
                                         A =  L + 2r X
where
       A = surface area, ft2
       L = width of front edge of mixing zone, ft
       r = radius of initial surface mixing, ft
       X = length of the mixing zone, ft
                                                                                 DRAFT

-------
                                             C-76

This approach will  characterize a proposed discharge by defining the volume of dilution  water per
unit time that will be required to achieve some acceptable concentration at the edge of the mixing
zone.   Also,  the length and width (and hence the surface area) of the necessary mixing zone will be
approximated.
C6.2      Sample Computations

The following computations are presented to illustrate the dilution volume method for a continuous
effluent discharge.

The following input values are used in the sample computations:

       Volume of effluent discharge per unit time Vp     = 44 cu ft/sec
       Turbulent dissipation parameter X               = 0.005
       Water column depth d                          = 10 ft
       Water velocity Vw                              =0.5 ft/sec
       Initial width of plume 2r                        = 30 ft
       Background concentration CBQ                   = 0. 1 mg/L
       Effluent discharge concentration Ce              = 30 mg/L
       Applicable water quality standard, C^           = 0.5 mg/L

The required volume per unit time will be:
                           = Vp D = 44
The required width of the mixing zone will be:
                                  __
                                   dVv    (10)  (0.5)
The time required to achieve the lateral spread  L  will be:
                                              3245           f
                                                                               DRAFT

-------
                                          C-77
                      t =    *    [(0.094) (649)2/3 -  (0.149) (15)2/3]
                                       - 1228 sec


The length of the mixing zone will be:       i

                         X =  (0.5  ft/sec) (1228 sec) = 614 ft


Thus the proposed mixing zone would have dimensions of:


                    Surface area  = /30 + 649| 614 = 208,453 sq ft

                        Maximum dimensions = 614 ft by 649 ft


This information would be used in considering  the compatibility of the  size of the mixing  zone
required to dilute the discharge with the available mixing zone.
                                                                          DRAFT

-------
                                           C-78

C7.0     REFERENCES

Ariathurai,  R., R.C. MacArthur and R.B. Krone.  1977.  Mathematical model of estuarial sediment
           transport.   Technical  Report D-77-12, U.S.  Army Engineer  Waterways  Experiment
           Station, Vicksburg, MS.

Bokuniewicz, H.I., et al.  1978.  Field study of the mechanics of the placement  of dredged material
           at open-water disposal  sites.  Technical Report D-78-7,  U.S.  Army Engineer Waterways
           Experiment Station, Vicksburg, MS.

Bowers, G.W. and M.K. Goldenblatt.  1978.  Calibration of a predictive model for instantaneously
           discharged dredged material.   U.S.  Environmental Protection Agency,  Corvallis, OR.
           EPA-699/3-78-089.

Brandsma,  M. G. and D.J. Divoky.  1976.  Development of models for prediction of short-term fate
           of dredged material discharged in  the estuarine environment.  Contract  Report D-76-5,
           DACW39-74-C-0075,  prepared  by Tetra Tech,  Inc.,  under  contract   to  U.S.  Army
           Engineer Waterways Experiment  Station, Vicksburg, MS.

Brandsma,  M. G. and T.C. Sauer, Jr.  1983. Mud discharge model: report and user's guide.  Exxon
           Production Research Co., Houston, TX.

Brooks, N. H.   1960.   Diffusion of sewage effluent in an ocean current.  Proceedings of First
           International Conference on Waste Disposal in the Marine Environment, Pergamon Press,
           NY, NY.

Buhler, J.  and W. Hauenstein.   1981.   Axismetric jets  in  a  crossflow.   19th Congress  of  the
           International Association of Hydraulic Research,  New Delhi, India, February 1-8, 1981.

Donekar, R.L. and Jirka, G.H.  1990. Expert System for  Hydrodynamic Mixing Zone Analysis of
           Conventional and Toxic  Submerged  Singe   Port  Discharges  (CORMIX1).    U.S.
           Environmental Protection Agency. PA/600/3-90/012.

Environmental Effects Laboratory.  1976. Ecological evaluation of  proposed discharge of dredged or
           fill  material  into  navigable  waters,  interim guidance for  implementation  of  Section
           404(b)(l)  of Public Law 92-500 (Federal Water Pollution Control  Act Amendments of
           1972).   Miscellaneous Paper  D-76-17, U.S.  Army Engineer  Waterways  Experiment
           Station, Vicksburg, MS.
                                                                             DRAFT

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                                            C-79

Fischer, H.  R. et al.   1979.   Mixing in Inland and  Coastal Waters.  Academic  Press, Inc., New
          York, NY.

Gowda, T. P.  H. 1984a.  Critical point method for mixing zones in rivers.  J. Environ. Engineer.,
          Am.  Soc. Civil Engineers  110: 244-262.

Gowda, T. P. H. 1984b. Water quality prediction in mixing zones of rivers.  J. Environ. Engineer.,
          Am.  Soc. Civil Engineers  110: 751-769.

Jirka, G. H. et al.  1981.  Buoyant surface jets. J. Hydraul. Engineer. Am. Soc. Civil Engineers
            107: 1467-1487.

Johnson, B. H.  1990.   User's  guide for  models of dredged  material  disposal in open  water.
          Technical Report D-90-5,  US Army Engineer Waterways Experiment Station, Vicksburg,
          MS.

Johnson, B.  and M. B. Boyd.   1975.  Mixing zone estimate for interior guidance.  Unpublished
          Memo.,  Mathematical Hydraulics  Division, Hydraulics Laboratory, U.S. Army Engineer
          Waterways Experiment Station, Vicksburg, MS.

Johnson, B.H., D.N. McComas, D.C. McVan and M.J. Trawle. 1994. Development and verification
          of  numerical models for predicting  the initial fate of dredged material  disposed in open
          water. Report 1, Physical model tests of dredged material disposal from a split-hull barge
          and a multiple bin vessel.  Draft  Technical Report,  U.S. Army Engineer Waterways
          Experiment Station, Vicksburg, MS.

King, I. P.  1992.  A  Finite Element Model for Stratified Flow-RMAlO-User's Guide-Version 4.3.
          Report prepared by Resource Management Associates for U.S. Army Engineer Waterways
          Experiment Station, Vicksburg, MS.

Koh, R.C.Y. and Y.C. Chang.  1973.   Mathematical model for barged ocean disposal  of waste.
          Environmental  Protection  Technology Series EPA 660/2-73-029, U.S. Army Engineer
          Waterways Experiment Station, Vicksburg,  MS.

Kraus,  N.C.   1991.  Mobile,  Alabama, field data collection project, 18 August-2  September 1989.
          Report 1, dredged material plume survey data report.   Technical Report DRP-91-3, U.S.
          Army Engineer Waterways Experiment Station, Vicksburg, MS.
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                                           C-80

Lin, H. J., Jones, N. L., and Richards, D. R.   1991.   A Microcomputer-Based  System for Two-
           Dimensional Flow Modeling. Proceedings of the 1991 National Conference on Hydraulic
           Engineering, ASCE, Nashville, TN.

Maclntyre, D. F.  1987.  Interim procedures for estimating mixing zones for effluent from dredged
           material disposal sites.  Technical Note  EEDP-04-5,  US  Army Engineer  Waterways
           Experiment Station, Vicksburg, MS.

Motz, L. H. and B. A. Benedict.  1972.  Surface jet model for heated discharges.  J. Hydraul. Div.,
           Am. Soc. Civil Engineers  98: 181-199.

Prakash,  A.  1977.  Convective-dispersion in  perennial  streams.  J. Environ. Engineer., Am. Soc.
           Civil Engineers 103: 321-340.

Schroeder, P.R.  and M.R. Palermo.   1990.     Automated  dredging  and disposal   alternatives
           management system, User's Guide.  Technical Note EEDP-06-12, U.S. Army Engineer
           Waterways Experiment Station, Vicksburg, MS.

Stefan, H.,  and J. S.  Gulliver.  1978.  Effluent mixing zone in a shallow river.  J. Environ.
           Engineer., Am. Soc. Civil Engineers  104:  199-213.

Thomas,  W.  A.,  and McAnally, W. H., Jr.   1990.  User's Manual for  the Generalized Computer
           Program System: Open-Channel Flow and Sedimentation, TABS-2. U.S. Army Engineer
           Waterways Experiment Station, Vicksburg, MS.

Wright, S. J.  1984. Buoyant jets in density-stratified crossflow.  J. Hydraul. Engineer., Am. Soc.
           Civil Engineers  110: 643-656.

Zeller, R. W., et al.  1971.  Heated surface jets  in steady crosscurrent.  J. Hydraul. Engineer., Am.
           Soc. Civil Engineers 97: 1403-1426.
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    APPENDIX D
STATISTICAL METHODS
                            DRAFT

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                               TABLE OF CONTENTS
                                                                                   Page No.
Table of Contents
List of Tables
List of Figures
 i
 ii
iii
Dl.O          INTRODUCTION	D-l
Dl.l          Basic Statistics	D-2
D1.2          Hypothesis Testing	D-5
D1.3          Experimental Design	D-7

D2.0          BIOLOGICAL EFFECTS  	D-8
D2.1          Tier III Water Column Toxicity Tests	D-9
D2.1.1         Comparison of 100% Elutriate and Dilution Water   	D-9
D2.1.1.1       Methods	D-9
D2.1.1.2       Analysis of Example Data	D-16
D2.1.2         Calculating Median Lethal Concentration	D-20
D2.1.2.1       Methods For Calculating LCj,	D-20
D2.1.2.2       Analysis of Example Data	D-24
D2.2          Tier III Benthic Toxicity Tests  	D-25
D2.2.1         Methods	D-25
D2.2.2         Analyses of Example Data	D-29

D3.0          BIOACCUMULATION	D-36
D3.1          Tier III Single-Time Point Laboratory Bioaccumulation Study  	D-36
D3.1.1         Comparisons with a Reference Sediment	D-37
D3.1.2         Comparisons with an Action Level 	D-43
D3.2          Tier IV Time-Sequenced Laboratory Bioaccumulation Study	D-46
D3.2.1         Calculating Steady-State Concentrations	D-47
D3.2.2         Comparison with Reference Sediments and Action Levels	D-51
D3.3          Steady-State Bioaccumulation from Field Data	D-S2

D4.0          SAS PROGRAMS AND OUTPUT FOR EXAMPLE DATA  	D-54
D4.1          Program WATTOX.SAS for Water Column Toxicity Test Data Analysis  	D-55
D4.1.1         WATTOX.SAS Program Statements 	D-55
D4.1.2         WATTOX.SAS Program Output  	D-57
D4.2          Program BENTOX.SAS for Benthic Toxicity Test Data Analysis	D-59
D4.2.1         BENTOX.SAS Program Statements	D-60
D4.2.2         BENTOX.SAS Program Output	D-63
D4.3          Program BIOACC.SAS for Single-Time Point Bioaccumulation Test Data Analysis .  D-68
D4.3.1         BIOACC.SAS Program Statements	D-68
D4.3.2         BIOACC.SAS Program Output	D-73
D4.4          Program  BIOACCSS.SAS for   Time-Sequenced Bioaccumulation Test  Data
              Analysis 	D-80
D4.3.1         BIOACCSS.SAS Program Statements 	D-81
D4.4.3         BIOACCSS.SAS Program Output	D-86

D5.0          REFERENCES	  D-102
                                                                            DRAFT

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                                    LIST OF TABLES
Table D-l.

Table D-2.

Table D-3.

Table D-4.


Table D-5.

Table D-6.

Table D-7.


Table D-8.


Table D-9.



Table D-10.
Types of Errors in Hypothesis Testing and Associated Probabilities.

Suggested a Levels to Use for Tests of Assumptions.
Page No.

     D-7

    D-12
Number of Survivors in a Hypothetical Water Column Toxicity Test After 96 h.      D-16
Tests of Assumptions and Hypothesis Tests on Arcsine-Transformed Water Column
Toxicity Test Example Data.

Calculated LC^ Values for Example Water Column Toxicity Test Data.

Number of Survivors in a Hypothetical Benthic Toxicity Test.

Tests of Assumptions and Parametric Tests of Hypotheses on Arcsine-Transformed
Benthic Toxicity Test Example Data.

Tests of Assumptions and Nonparametric Hypothesis Tests on Benthic Toxicity Test
Example Data Converted to Rankits and Ranks.

Results from a Hypothetical Single-Time Point Bioaccumulation Test, Showing
Contaminant  Concentrations  (ng/g) in Tissues of Animals Exposed to Different
Treatments.

Tests of Assumptions and Parametric Hypothesis Tests on Untransformed and
Log10-Transformed Bioaccumulation Example Data.
Table D-ll.    Tests of Assumptions and Nonparametric Hypothesis Tests on Bioaccumulation
               Example Data Converted to Rankits and Ranks.

Table D-12.    Results from a Hypothetical  Time-Sequenced  Bioaccumulation  Test, Showing
               Contaminant Concentrations (jig/g) in  Tissues of Animals Exposed to Different
               Treatments.

Table D-13.    Regression Parameters Estimated from Example Time-Sequenced Bioaccumulation
               Data.

Table D-14.    Tests of Assumptions and Parametric Hypothesis Tests on Untransformed Steady-
               State Bioaccumulation Example Data.
    D-18

    D-25

    D-30


    D-33


    D-34



    D-37


    D-41


    D-42



    D-48


    D-49


    D-53
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                                              Ill
                                   LIST OF FIGURES
                                                                                      Page No.

Figure D-l.    Water Column Toxicity Test Decision Tree.                                     D-17

Figure D-2.    LC,, Decision Tree.                                                          D-21

Figure D-3.    Probit Plot of Water Column Toxicity Test Example Data.                        D-26

Figure D-4A.   Benthic Toxicity Test Decision Tree (Parametric Tests).                           D-31

Figure D-4B.   Benthic Toxicity Test Decision Tree (Nonparametric Tests).                       D-32

Figure D-5A.   Bioaccumulation Test Decision Tree (Parametric Tests).                          D-38

Figure D-5B.   Bioaccumulation Test Decision Tree (Nonparametric Tests).                       D-39

Figure D-6    Comparison of Mean Dredged Sediment Contaminant Tissue Levels (mean) and 95%
              Upper Confidence Level (UCL) with Hypothetical Action Level.                   D-44

Figure D-7.    Comparison of Mean Dredged Sediment Contaminant Steady-State Tissue Levels
              (CM) (mean) and 95%  Upper Confidence Levels (UCL) with Hypothetical Action
              Level.                                                                     D-53

Figure D-8.    Plot of  Time-Sequenced Bioaccumulation Reference  Sediment Example Data by
              Replicate.                                                                  D-89

Figure D-9.    Plot of  Time-Sequenced Bioaccumulation Dredged Sediment 1 Example Data by
              Replicate.                                                                  D-90

Figure D-10.   Plot of  Time-Sequenced Bioaccumulation Dredged Sediment 2 Example Data by
              Replicate.                                                                  D-91

Figure D-ll.   Plot of  Time-Sequenced Bioaccumulation Dredged Sediment 3 Example Data by
              Replicate.                                                                  D-92
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	D-l	

APPENDIX D - STATISTICAL METHODS

Dl.O           INTRODUCTION

This Appendix presents the appropriate statistical methods for analyzing data from toxicity and bioaccumula-
tion tests. The methodology is not intended to be exhaustive, nor is it intended to be a "cook-book" approach
to data analysis. Statistical analyses are routine only under ideal experimental conditions.  The methods
presented here will usually be adequate for the tests conducted under the conditions specified in this
document.  An experienced applied statistician should be consulted whenever there are questions.

The following are examples of departures from ideal experimental conditions that may require additions to
or modifications of the statistical methods presented in this chapter:

        •      Unequal numbers of experimental animals assigned to each treatment container, or loss of
               animals during the experiment

        •      Unequal numbers of replications (i.e., containers or aquaria) of the treatments

        •      Measurements scheduled at selected time intervals actually performed at other times

        •      Different conditions of salinity, pH, dissolved oxygen, temperature, etc., among exposure
               chambers

        •      Differences in placement conditions of the testing containers, or in the animals assigned to
               different treatments

        •      Contaminant concentration data reported as less than detection limit.

Problems such as these, which result in non-ideal data, will be examined and illustrated in detail in an Appli-
cations Guide to be developed by the USAGE as a supplement to this Appendix.

The following statistical methods will be  presented as each applies to a specific test procedure:

        •      Tests of assumptions (normality and equality of variances)

        •      Data-scale transformations

        •      Two-sample Mest

        •      Nonparametric two-sample test
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	D-2	

        •       Power and sample size calculations

        •       LC,0 calculations

        •       Parametric multiple comparisons among treatments

        •       Nonparametric multiple comparisons among treatments

        •       Confidence interval calculations

        •       Comparisons to action levels

Decision trees are included to provide a general overview of each biological test. These trees illustrate which
of the above statistical methods are appropriate for analyzing the results of each biological test, and the order
in which the statistical procedures should be conducted.  The trees include three general levels of decisions
in the biological testing evaluation process: (1) decisions made by evaluating the experimental  QA/QC and
examining dredged material and reference means, (2) decisions concerning which statistical comparison proce-
dure to use based on tests of assumptions, and (3) decisions concerning the significance of statistical compari-
sons.

The statistical methods (with the exception of LC50 procedures) are illustrated in this Appendix with example
data analyzed by SAS IBM-compatible PC programs (SAS Institute, Inc., 1988a-c).  This manual does not
constitute official endorsement or approval of these or any other commercial hardware or software products.
Other equally acceptable hardware and software products are  commercially available and may be  used  to
perform the necessary analyses. For example, all analyses required for this Appendix can be conducted using
SYSTAT (Steinberg, 1988; Wilkinson, 1990; Steinberg and Colla, 1991), with different tests for normality and
equality of variances. If it is necessary to write original programs to perform statistical  analysis, the appro-
priateness of the techniques and accuracy of the calculations must be very carefully verified and documented.

Each example data set included in this Appendix is analyzed using several different statistical methods (usually,
all of the possible tests in the appropriate decision tree) for illustrative purposes only. Note that the results
of different statistical tests will occasionally disagree, and it is never appropriate  to conduct several tests in order
to choose the result one likes best.  Decisions concerning the proper statistical tests to  use should be made a
priori, based on such considerations as experimental design, hypotheses of interest, relative importance of Type
I and Type II error rates (Section  D1.2), and tests of assumptions (Section D2.1.1.1).


Dl.l            Basic Statistics

Statistical methods are used to make inferences about populations, based on samples from those populations.
In most toxicity and bioaccumulation tests, samples of exposed organisms are used to estimate the response
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	D-3	

of the population of laboratory organisms.  The response from the samples is usually compared with the
response to a reference1, or with some fixed standard  such as an FDA action level.  In any toxicity or
bioaccumulation  test, summary  statistics such as means and standard errors  for response variables (e.g.,
survival, contaminant levels in tissue) should be provided for each treatment (e.g., elutriate concentration,
sediment).

In the tests described herein, samples or observations refer to replicates of treatments. Sample size n is the
number of replicates (i.e., experimental units, test containers) in an individual treatment, not the number of
organisms in a test container. Overall sample size N is the total number of replicates in all treatments com-
bined, i.e.,

        N       =      nl + n2 + n3 + ... + nk

where k is the total number of treatments in the experiment.

The statistical methods discussed in this Appendix are described in general statistics texts such as Steel and
Torrie (1980), Sokal and Rohlf (1981), Dixon and Massey (1983), Zar (1984), and  Snedecor  and Cochran
(1989). We recommend that investigators using this Appendix have at least one of these texts on hand.  A
nonparametric statistics text such as Conover (1980) can also be helpful.

Mean

The sample mean (x)  is the average value, or Hr, / n, where

        n       =      number of observations  (replicates)
        jtj       =      ith observation, e.g., x2 is the second observation
        SXj      =      every x summed  = xl + x2 + x3 + .. . + xn; usually written  Sx

Most calculators and statistical  software packages will provide means.
Standard deviation

The sample standard deviation (5) is a measure of the variation of the data around the mean.  The sample
variance, s\ is given by:
    1    Reference is  used generically to refer either to a reference sediment (as in benthic toxicity and
        bioaccumulation testing), or to dilution water or control water (used in water column toxicity testing).
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                                                D-4
                                                n -  1

Standard error of the mean
                                                                                           (Eq.
The standard error of the mean (SE, or shTri) estimates variation among sample means rather than among
individual values. The SE is an estimate of the SD among means that would be obtained from several samples
of n observations each.  Most of the statistical tests in this manual compare means with other means (e.g.,
dredged sediment mean with reference mean) or with a fixed standard (e.g., FDA action level). Therefore,
the "natural" or "random" variation of sample means (estimated by SE),  rather than the variation among
individual observations (estimated by s), is required for the tests.

In addition to the summary statistics above, two other statistics derived from the normal (bell-shaped) fre-
quency distribution are central to statistical testing and to the tests described in this Appendix. These two
statistics are normal deviates (2-scores) and Student's t.

Normal deviates (z)

Z-scores or normal deviates measure distance from the mean  in  standard deviation units in a  normal
distribution.  For example, a point 1 standard deviation greater than the mean has a  z-score of 1; the mean
has a z-score of 0. Z-scores are usually associated with a cumulative probability or proportion. For example,
suppose an investigator wants to know the proportion of values in a normal distribution less than or equal
to the mean plus 1 standard deviation. In this situation z=0.84, i.e., in a normal distribution 84% of values
will be less than or equal to the mean plus 1 standard deviation. Alternatively, an investigator may want to
determine the z-score associated with a specific proportion or probability.  For example, he or she may want
to know the range in which 95% of the values in a normal distribution should fall. That range is the mean
± 1.96 standard deviation (z-scores from -1.96 to +1.96).

Tables of z-scores can be found in most statistical texts, and bear titles such as  "Standard Normal Cumulative
Probabilities," "Ordinates of the Normal Curve," or "Normal Curve Areas." Typically the z-scores are listed
in the column (top) and row (left) margins, with the column marginal value being added to the row marginal
value to obtain the z-score.  The body  of the table contains the probability associated with each z-score.
However, depending on the table, that probability may refer to the proportion of all values less than the z-
score, the proportion of values falling between 0 and the z-score, or the proportion of values greater than the
z-score. For example, if the z-score is 1.96,97.5% of the values in a normal distribution fall below the z-score
(Kleinbaum and Kupper, 1978, Table A-l), 47.5% fall between 0 and the z-score (Rohlf and Sokal, 1969, Table
P), and 2.5% fall above the z-score (Steel and Torrie, 1980, Table A4).  It is important to distinguish which
probability is of interest.

Z-scores can also be obtained from functions in statistical software packages.  For example, in SAS the
PROSIT function will return a z-score  for a specified probability, and the PROBNORM function will compute
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                                                D-5

the proportion of values less than a given z-score.

Student's t

Normal deviates can only be used to make inferences when the standard deviation is known, rather than esti-
mated.  The true population mean (p.) and standard deviation (a) are only known if the entire population is
sampled, which is rare.  In most cases samples are taken randomly from the population, and the s calculated
from those samples is only an estimate of o.  Student's r-values account for this uncertainty, but are otherwise
similar to normal deviates. For example, an investigator may want to determine the range in which 95% of
the values  in a population should fall, based on a sample of 20 observations from that population. If  the
sample consisted of the entire population, /i and a would be known with certainty, and normal deviates would
be used to  estimate the  desired range (as in  the above paragraph).  However, if the sample represented only
a small  proportion of the population, r-values would be used to estimate the desired  range.  The degrees of
freedom for the  test, which is defined as the  sample size minus one (n-1), must be used to obtain the correct
r-value.  Student r-values decrease with increasing sample size, because larger samples provide a more precise
estimate of/i and o. For a probability of 95%, the appropriate range of r-values is -2.09 to +2.09.  In other
words, 95% of the values in the population should lie within the range: sample mean ±2.09 s. Note that this
is wider than the corresponding range calculated using normal deviates.  As sample  size increases, r-values
converge on the z-scores for the same probability.

Tables of r-values typically give the degrees of freedom (df or v) in the row (left) margin and probabilities or
percentiles in the column (top) margin.  Percentiles refer to the cumulative proportion of values less than r,
whereas probabilities (also known as a in this case) refer to the proportion of values less than -r and/or greater
than +r. A two-tailed probability refers to  both "tails"  of the r-distribution curve, i.e., the probability of a
value either >+r or <-r. A one-tailed probability refers to only one of the tails of the curve, e.g., the proba-
bility of a value  >+r.

When using a r table, it is crucial to determine whether the table is based on one-tailed probabilities (such
as Table V in McClave  and Dietrich, 1979,  and Table A-2 in  Kleinbaum and  Kupper, 1978), or two-tailed
probabilities (such as Table A.3 of Steel and Torrie, 1980). Some tables give both (such as Table B.3 of Zar,
1984).  For most applications involving  r-values in this Appendix, one-tailed probabilities are desired. The
body of the table contains the r-value for each df and percentile (or a). The r-value for a one-tailed probability
may be found in a  two-tailed table by  looking up r under the column  for  twice the desired one-tailed
probability. For example, the one-tailed r-value for  a = 0.05 and df = 20 is 1.725, and is found in a two-tailed
table using the column for a = 0.10.

Statistical software packages may also provide functions to determine r-values or their associated probabilities.
In SAS, these functions  are TINV and PROBT.
D1.2           Hypothesis Testing
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	D-6	

The goal  in analyzing toxicity and bioaccumulation test data is to determine whether the mean effect of
exposure to a dredged sediment is significantly greater than the mean effect of exposure to a reference.  Two
formal hypotheses underlie the statistical analysis of data in the two-sample situation. Let/uR denote the mean
effect of exposure to the reference R and let JID denote the mean effect of exposure to the dredged sediment
D.  Then, these two hypotheses are defined as follows:

        Null hypothesis

Case 0:         HO: /ID = /IR
                        There is no difference in mean effect between the  treat-
                        ment (dredged sediment) and reference.

        Alternative hypotheses

Case 1:         H^ /ID < /UR

                        The mean effect of the dredged sediment is less than the
                        mean effect of the reference (e.g., survival).

                                                 OR

Case 2:         H^ j*D > /iR

                        The mean effect of the dredged sediment  is greater than
                        the mean effect of the reference (e.g., bioaccumulation).

Our hypothesis test will either reject H0 for Ht (Case 1 or Case 2), or will be unable to  reject HO (Case 0).
A one-tailed test is used because there is little concern about identifying a lesser negative effect from the
dredged sediment than from  the reference.

In performing the hypothesis test, and in determining the sample size to use in the test, the investigator must
be aware of the probabilities for two types of errors that can occur in the conclusion.  Type I errors occur if,
after analysis of the data, H0 is rejected when it was actually true. In Case 1 for example, a  Type I error occurs
when it is concluded that the mean effect (e.g., survival) of the dredged sediment is less than the mean effect
of the reference when, in fact, the true mean effect of the dredged sediment is not less than that for the
reference.  Type II errors occur when H0 is not rejected when it actually should  have been rejected (e.g., in
Case 2, it is concluded that there is no difference in mean effects of the dredged sediment and reference when,
in fact, the true mean effect of the dredged sediment is greater than that of the reference).

To be environmentally protective in dredged  sediment disposal evaluations,  it is more important to guard
against Type II errors.  A Type II error could result in inappropriate placement  of dredged sediment in the
aquatic environment, while a Type I error could result in more costly  alternatives to aquatic disposal. The
probability of a Type I error is often represented by the letter a; the probability of a Type II error is often
written as p. The significance level or confidence level of a statistical test is 1 - a.  The power of a test is 1 -
 P, which is the probability of rejecting H0 when it should be rejected, or in other words, the power to detect
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	D-7	

true significant differences.  For example, in Case 2 above, the power is the probability of concluding that the
mean effect is greater in the dredged-sediment group when, in fact, this is true.  The types of errors and their
associated probabilities are summarized in Table D-l.

Table D-l.      Types of Errors in Hypothesis Testing and Associated Probabilities.
Hypothesis Test
Conclusion
H« True
(do not reject)
HO False
(reject)
True State of Nature
HO True
Correct
(probability = 1 - a)
Type I Error
(probability = a)
HO False
Type II Error
(probability = P)
Correct
(probability = 1 - p)
In hypothesis testing, the Type I error rate is usually prespecified (biological tests, by convention, generally
set a = 0.05, although there is nothing magical about this probability).  An ideal statistical procedure for
hypothesis testing seeks to maintain the predetermined a, while minimizing the Type II error rate (i.e., maxi-
mizing power).  It  may not be possible to do both, particularly if the sample data  depart from a normal
distribution.  A test that does well in maintaining the predetermined a, regardless of the characteristics of the
sample data, is considered "robust." Tests included in this Appendix were chosen primarily on the basis of
power rather than robustness, as the consequences of Type II error were considered more severe than those
of Type I error.

Simple formulae for calculating the power of the statistical  tests used in this Appendix are presented along
with the descriptions of the tests in Sections D2.1.1.1, D2.2.1, D2.2.2, D3.1.2, and D3.2.2. The formulae may
be used to calculate the sample size required to ensure a specific power of detecting an effect of a given magni-
tude (effect size), assuming that effect  exists.  The  formulae can  also be used to calculate the power  of a
specific sample  size to detect a specified difference.  This latter approach is often more relevant than
calculating required sample sizes because budget or logistical constraints usually limit the number of replicates
that can be used in biological tests. This is especially true if the tests include expensive chemical analyses (e.g.,
Tiers III and IV bioaccumulation tests).
D1.3            Experimental Design

Once the investigator has formulated the null hypotheses to be tested, decided upon significance (a) and power
(1-P) levels for hypothesis testing, and determined the sample size necessary to achieve the desired power, the
next step is to design an experiment  to  test the hypotheses.  Instructions for setting up and conducting
sediment toxicity and bioaccumulation  experiments are outlined in Chapters 11 and 12, but it is important at
this  point to review the basic principles of experimental design.  These  principles include replication,
randomization, interspersion, and controls (Hurlbert, 1984).
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	D-8	

Replication refers to the assignment of a treatment to more than one experimental unit.  The number of
replicates, as stated earlier, is the sample size for that treatment. Recall that an experimental unit or replicate
is the test container (e.g., a  beaker or an aquarium), not an individual organism in the test container.  The
number of organisms in the test container is important only in terms of constituting an adequate measure of
the endpoint  being tested  (e.g., providing sufficient  tissue to  measure contaminant  bioaccumulation).
Replication of treatments is necessary to control for random error in the conduct of the experiment.  Appendix
E includes guidelines for minimum number of replicates for various Tier III and IV bioassays.  However, we
strongly recommend determining sample size a priori using the power formulae  in Sections D2.1.1.1, D2.2.1,
D2.2.2, and  D3.2.2. In many cases, the number of replicates necessary for a powerful statistical test will be
greater than the minimum guidelines.

Randomization and interspersion refer to the actual placement of experimental units in the laboratory setup.
A random numbers table, available in most statistical texts, may be  used to randomly assign treatments to the
experimental units. If the randomization does not achieve a reasonable interspersion of treatments, e.g. if
several experimental units of the same treatment are clumped together, then a new randomization should be
tried.  Randomization and interspersion are necessary to control for investigator bias, for initial or inherent
variability among experimental units, and for variability in environmental conditions such as lighting, water
flow, etc.

Replication, randomization,  and interspersion all function to control extraneous sources of variability in an
experiment. In addition, control treatments) are needed to control  temporal or procedural variability.  In the
broadest sense, the control treatment is simply the treatment against which the other treatments are compared.
This is the dilution water (or control water) in water column toxicity testing, and the reference sediment in
benthic toxicity and bioaccumulation  testing.  Laboratory controls, such  as  a  clean  sand exposure in
bioaccumulation testing, may also be included.  In Tiers III and IV testing, laboratory controls are used for
quality assurance, and are not included in the statistical analyses.

Testing in Tiers III and IV can in most cases be best accomplished using simple experimental designs, either
a completely randomized design or a randomized complete blocks design. These designs are discussed in most
general statistics texts.  In a completely randomized design, treatments are assigned to experimental units
randomly over the entire experimental setup. A randomized complete blocks design should be used when the
experimental units are placed on or in several different tables, benches or water baths (i.e., "blocks").  Each
block holds a certain proportion of the experimental units.  Treatments are assigned to experimental units
randomly within each block,  and each block contains an equal number of replicates of each treatment. Either
of these designs  is acceptable, providing the  principles  of  replication, randomization,  interspersion, and
controls are followed.  Adherence to  the principles of  experimental design ensures  that the most basic
assumption  of statistical hypothesis testing, the assumption that treatments are sampled independently, is met.
D2.0           BIOLOGICAL EFFECTS
                                                                                     DRAFT

-------
	D-9	

D2.1           Tier III Water Column Toxicity Tests

The objective of the analysis of Tier III water column toxicity test data is to assess the evidence for reduced
survival due to toxicity of suspended  plus dissolved dredged sediment constituents.  If reduced survival is
evident, then the median lethal concentration (LC50) or effective sublethal concentration (EC^) of the dredged
sediment is calculated from the serial dilution experiment described in Section 11.1.4. Figures D-l and D-2
provide an overview of water column toxicity test data analysis. Control survival must be *90% or some other
appropriate value, otherwise the test must be repeated (Section 13.3.17.3). At the end of the exposure period,
the effects, if any, on the survival of the test organisms should be clearly manifest in the 100% elutriate con-
centration. When the dilutions are prepared with other than control water, the dilution water treatment is
preferred over the control water for the data analysis.  If the elutriate survival exceeds the control survival,
then the toxicity test indicates no adverse impact from the dredged sediment (Section 11.1.5).
D2.1.1          Comparison of 100% Elutriate and Dilution Water

1)2.1.1.1         Methods

Two-sample Mest

The usual statistical test for comparing two independent samples such as the 100% elutriate and the dilution
water is the two-sample Mest (Snedecor and Cochran, 1989). The Mest will also be used  in some circum-
stances in benthic toxicity and bioaccumulation tests, to compare individual dredged sediments with a reference
(see Figures D-l, D-4A, D-5A).

The /'-statistic for  testing  the equality  of means x^ and x2  from two independent samples with nt and «2
replicates is:
                                                                                            (Eq< 2)
where s^,^, the pooled variance, is calculated as:


                                                       D] /(«!+«,- 2) .                   <** 3)
and where si and si are the sample variances of the two groups.  If the sample sizes are equal (n1 = «2), then:
                                                                                     DRAFT

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                                               D-10
The calculated t is compared with the Student t distribution with nl + n2-2 degrees of freedom.

The use of Eq.2 to calculate t assumes that the variances of the two groups are equal.  If the variances are
unequal (see Tests for Equality of Variances below), t is computed as:
t = (*, - xj) / Jfa + fa .
                                  (^/n,)2 / («, - 1) + (s>2)2 / (rtj - 1)
                                                                                                5>
This statistic is compared with the Student t distribution with degrees of freedom given by Satterthwaite's
(1946) approximation:
                                                                                            (Eq.6)
This formula can result in fractional degrees of freedom, in which case one should round df down to the
nearest integer in order to use a t table. The degrees of freedom for the r-test for unequal variances will
usually be less than the degrees of freedom for the r-test for equal variances.

Tests of Assumptions

The two-sample f-test for equal variances (and other parametric tests such as analysis of variance) is only
appropriate if:

        •       there are independent, replicate experimental units for each treatment,

        •       the observations within each treatment follow a normal distribution, and

        •       variances for both treatments are equal or similar.

The first assumption is an essential component of experimental design (Section D1.3). The second and third
assumptions can be tested using the data obtained from the experiment. Therefore, prior to conducting the
r-test, tests for normality and equality of variances should be performed. In some statistical software packages,
these tests of assumptions are done in conjunction with Mests  or as  part of data summary or screening
routines  that also provide means, s, SE and various diagnostic statistics.
                                                                                     DRAFT

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	D-ll	

Outliers (extreme values) and systematic departures from a normal distribution (e.g., a log-normal distribution)
are the most common causes  of departures from normality and/or equality of variances. An appropriate
transformation will normalize many distributions. In fact, the arcsine transformation (arcsine, in radians, of
•fp, where/? is the survival expressed as a proportion) is so effective, and so frequently necessary, that this
Appendix recommends applying it automatically to all survival data in the analysis of toxicity tests. Problems
with outliers can usually be solved only by using nonparametric tests, but careful  laboratory practices can
reduce the frequency of outliers.
Tests for Normality

The most commonly used test for normality for small sample sizes (
-------
                                               D-12
Table D-2.     Suggested a Levels to Use for Tests of Assumptions.
Test
Normality
Equality of
Variances
Number of
Observations*
N = 3 to 9
N = 10 to 19
N = 20 or more
n = 2 to 9
n = 10 or more
a When Design Is
Balanced
0.10
0.05
0.01
0.10
0.05
Unbalanced6
0.25
0.10
0.05
0.25
0.10
 1N = total number of observations (replicates) in all treatments combined; n = number of observations
     (replicates) in an individual treatment
 b n^ * 2nmin

Tables of quantiles of W can be found in Shapiro and Wilk (1965), Gill (1978), Conover (1980), USEPA
(1989) and other statistical texts.  These references also provide methods of calculating W, although the calcu-
lations can be tedious. For that reason, computer programs are preferred for the calculation of W.  SAS can
calculate Pausing the NORMAL option in PROC UNIVARIATE (see Program WATTOX.SAS in Section
D4.1).

The Kolmogorov-Smirnov (K-S) Test is also an acceptable test for normality for small sample sizes, provided
that the probabilities developed by Lilliefors (1967) are used (Sokal and Rohlf, 1981). The SYSTAT NPAR
module provides the appropriate test, and specifically identifies the test as Lilliefors Test (Wilkinson, 1991).
Other statistical packages providing K-S Tests may not use the Lilliefors probabilities, and the package
documentation should always be checked to determine if the appropriate probabilities are provided. The chi-
square (x2) test for normality can be used for larger sample sizes (e.g., N > 50) (Sokal and Rohlf, 1981).

Tests for Equality of Variances
There are a number of tests for equality of variances. Some of these tests are sensitive to departures from
normality, which is why a test for normality should be performed first.  Bartlett's Test, Levene's Test, and
Cochran's Test (Winer, 1971; Snedecor and Cochran, 1989) all have similar power for small, equal sample sizes
(n=5) (Conover et al., 1981), and any one of these tests is adequate for the analyses in this Appendix.  Many
software packages for Mests and analysis of variance (ANOVA) provide at least one of the tests.  Levene's
Test can easily be performed by comparing the absolute values of residuals between treatments using r-tests
or ANOVA. SAS statements for conducting Levene's Test are provided in BENTOX.SAS, BIOACC.SAS and
BIOACCSS.SAS programs (Sections D4.2.1, D4.3.1 and D4.4.1).

If no tests for equality of variances are included in the available statistical software, Hartley's F^ can easily
                                                                                   DRAFT

-------
	D-13	

be calculated:
                            f^ =  (larger of *i. sl) / ( smaller of si, si)

When FMV is large, the hypothesis of equal variances is more likely to be rejected. F^ is a two-tailed test
because it does not matter which variance is expected to be larger.  Some statistical texts provide critical values
°f Fma (Winer, 1971; Gill, 1978 [includes a table for unequal replication, but only for a  = 0.05]; Rohlf and
Sokal, 1969).  In the two-sample case, Hartley's F^ is the same as the Folded-F or F' test. The F' test is
conducted automatically in the SAS TTEST procedure.

Cochran's Test, where C = the largest variance divided by the sum of the variances, is also simple to calculate
by hand, and is somewhat more powerful  then Hartley's F,^, for small, equal sample sizes (Conover et al.,
1981). However, tables of critical values of Cochran's C are not available in most statistical texts.  Winer
(1971) and Dixon and Massey (1983) include a table for Cochran's Test, but the tables are limited to tests with
equal sample sizes. Tables of critical values for tests such as Cochran's  C and Hartley's F^ may  also be
restricted to one or two a levels (usually 0.05 and 0.01). Because of the limitations of these tables, computer
programs are preferred for tests of equality of variances.

Levels of a for tests of equality of variances are provided in Table D-2; these depend upon number of repli-
cates in a treatment (n) and allotment of replicates among treatments (design).  Relatively high a's are recom-
mended because the power of the above tests for equality of variances is rather low (about 0.3) when  n is
small.  Equality of variances  is rejected if the probability associated with the test statistic is less than the
appropriate a.  If the test for equality of variances is significant  even after transformation, the /-test for
unequal (separate) variances should be selected rather than the r-test for equal (pooled) variances.

Nonparametric Tests

Tests such as the /-test, which analyze the original or transformed data, and which rely on the properties of
the normal distribution, are referred to as parametric tests. Nonparametric tests, which do not require  that
data be normally distributed,  analyze the ranks of data, and generally compare medians rather than  means.
The median of a sample is the middle or 50th percentile observation when the data are ordered from smallest
to largest.  In many cases, nonparametric tests can be performed simply by converting the data to ranks or
normalized ranks, and  then conducting the usual parametric test procedures on the ranks.

Nonparametric tests are  useful because  of their  generality, but may  have less  statistical  power than
corresponding parametric tests when the parametric test assumptions are met.

When parametric tests  are not appropriate for comparisons because the normality assumption is not met, we
recommend converting the data to normalized ranks (rankits). Rankits are simply the z-scores expected for
the rank in a normal distribution.  Thus, using rankits imposes a normal distribution over all the data, al-
though not necessarily  within each treatment.  Rankits can be obtained by ranking the data, then converting
the ranks to rankits using the following formula:
                                                                                     DRAFT

-------
                                               D-14
                                    rankit = *!(«»*- 0375) IV,. (US)]'                            
where z is the normal deviate and W is the total number of observations. For example, the approximate rankit
for the sixth lowest value (rank=6) of 20 would be z[(<.a375)/(20 + a25>]» which is za27g or -0.59.

In SAS, normalized ranks or rankits can be provided in PROC RANK with the NORMAL=BLOM option.
In SYSTAT and other packages, the ranks must be converted to rankits using the formula above (the conver-
sion is a one-line command).  In some programs the conversion may be more difficult to  make, especially if
functions to provide z-scores for any probability  are not available. When rankits cannot easily be calculated,
the original data may be converted to ranks.

In comparisons involving only two treatments, there is no real need to test assumptions on the rankits or
ranks; simply proceed with a one-tailed Mest for unequal variances using the rankits or ranks.

Statistical Power

For a Mest, the basic formula for calculating the sample size (number of replicate experimental units, n) per
treatment necessary to provide a specified  power (1-p) to detect a given effect size (d) is:
where  v = degrees of freedom (df) or (n1 + «2 - 2)
        *!.., = Student f-value for probability 1-a and v df
        ft.j₯ = Student f-value for probability 1-p and v df
        d = the effect size or difference to be detected.

Recall that p is the probability of committing a Type II error.  This formula for n must be solved iteratively,
because an initial value of n must be used to determine v. A new n is then calculated using the initial value,
and the process is repeated until n and v  are consistent. The iterative process can be tedious  if computer
programs are not used. It is easier to use  the following approximate formula (from Alldredge, 1987):
                                 = 2 &-  + Z-  )2 (*2#2> + 0-25(2-) ,                       <**»• 9>
where  za.« = normal deviate for 1-a
        Zj.0 = normal deviate for 1-p
        0.25(zf..) = correction term to increase sample size when n is small
                                                                                     DRAFT

-------
	D-15	

Calculated n derived from this formula should be regarded as approximate for n<5.  Regardless of which
formula is used, a fractional n is always rounded up to the next integer.

A useful exercise when sample sizes are fixed because of budget or logistic constraints is to calculate the power
of the test to detect a specific effect size (d). In a test comparing 100% elutriate survival with dilution water
survival, d is some selected reduction in mean 100% elutriate survival from mean dilution water survival.  Eq.
8 can be rearranged and solved for t^t to determine the power:
We then enter a / table at v df and find the column closest to the value of r,.p; power * l-P, where P is the
probability for that column.  SAS can calculate power more exactly using the PROBT function for t^ and v
df. Note that f-values can be used because both n and v are known. One can also calculate the difference that
can be detected for any given power and sample size:
                                                                                                U>
The simplest power to use is 0.50, because then ft.p=0. Many computer programs will provide this difference,
usually referred to as the "minimum significant difference", "least significant difference" or some similar term.
The term "average detectable difference" would also be applicable, as this is the difference we expect to be able
to detect 50% of the time. In this Appendix, we recommend reporting the minimum significant difference or
some other indication  of power along with the results of statistical analyses.  If power is consistently and
regularly reported, investigators will gain an appreciation of the strengths and limitations of various toxicity
tests and analyses.

If values are transformed prior to analyses, all power calculations should be done on the transformed scale.
In the case of arcsine-transformed survival, a constant effect size d on the percentage or proportion scale will
not be constant on the arcsine scale, because the latter scale spreads out high and low values. Therefore, a
reference  survival must  be  specified and arcsine-transformed, and the effect size  also transformed to a
difference on the arcsine scale.  For example, suppose we wanted to calculate the power of a Mest to detect
a 25% reduction in survival from the reference. A reasonable reference survival (e.g., 90%) would be specified
and arcsine-transformed (=1.249). We would also arcsine-transform a 25% reduction (=65% survival or 0.938
after transformation).  The difference d would then be 1.249 - 0.938 or 0.311,  and that value would be used
in power calculations. Experimentation with arcsine-transformed data will rapidly reveal that toxicity  tests are
more powerful, in terms of the size of differences that can be detected on the original (untransformed) scale,
when reference survival is higher. In other words, we are more likely to detect a 25% reduction in survival
                                                                                     DRAFT

-------
	D-16	

if reference survival is 90% than if reference survival is 75%. This is precisely what happens in real toxicity
tests, which is why the arcsine transformation is used for survival data.

Simple formulae for calculation of sample  size or power  are not available for the tests of assumptions
recommended in this Appendix.
D2.1.1.2
Analysis of Example Data
Table D-3 contains example data from a 96-h water column toxicity test using a dilution water and a dredged-
sediment elutriate at four serial dilutions. In this example, control (laboratory) water was also used for
dilutions, and no separate control was necessary. In other cases, the dilution water may be receiving water and
a separate laboratory control would be required. Analysis of this example data will be conducted using the
decision tree in Figure D-l. Numbers in parentheses in the text refer to numbered nodes of the decision tree.
The SAS program WATTOX and complete results for water column toxicity test data analyses are provided
in Section D4.1; some additional analyses were conducted using SYSTAT programs.

Means (1) and SE for the survival data are provided in Table D-3.  Overall mean survival in the control (=
dilution) water was 98%, indicating that the test was acceptable (2).  The statistical comparison of 100%
elutriate survival and dilution water survival was then conducted because the 100% elutriate survival was at
least 10% lower than the dilution water survival (3). The next step was to arcsine-transform the survival
proportions for the dilution water and 100% elutriate treatments (4).

Table D-3.     Number of Survivors  in a Hypothetical Water Column Toxicity Test After 96 h.
Replicate*
1
2
3
4
5
Total
Mean
SE
Treatment8
Dilution Water0
20
19
20
20
19
98
19.6 (98%)
0.24
100%
6
7
9
5
8
35
7.0 (35%)
0.71
50%
8
8
9
10
11
46
9.2 (46%)
0.58
25%
12
18
15
14
13
72
14.4 (72%)
1.03
12.5%
17
17
18
16
18
86
17.2 (86%)
0.37
* Percent concentrations of dredged-material elutriate:   b
100% =   1 part elutriate plus 0 part dilution water
50% =    1 part elutriate plus 1 part dilution water     c
25% =    1 part elutriate plus 3 parts dilution water
12.5% =   1 part elutriate plus 7 parts dilution water
                                              20 organisms per replicate at initiation of
                                              test
                                              In this example, the dilution water was
                                              control (laboratory) water
                                                                                     DRAFT

-------
                                              D-17
©
©
                   SURVIVAL DATA -
             100% ELUTRIATE VS. DILUTION
 ARCSINE TRANSFORM
                         1
          ARE  DATA NORMALLY DISTRIBUTED?
                ARE VARIANCES EQUAL?
ONE-TAILED T-TEST FOR
  EQUAL VARIANCES
                                                NO
                  CALCULATE MEANS
                 IS CONTROL WATER
               SURVIVAL 2: STANDARD?
                 IS SURVIVAL IN 100%
         ELUTRIATE AT LEAST 10% LESS THAN
              DILUTION WATER SURVIVAL?
                                                      REPEAT TEST
                  IS SURVIVAL IN 100%
                ELUTRIATE SIGNIFICANTLY
           LOWER THAN IN DILUTION WATER?
                      YES
         T
                  PROCEED TO LC50
              CALCULATIONS (FIGURE D-2)
                                                       DREDGED MATERIAL
                                                        MEETS GUIDELINES
                                                       FOR WATER COLUMN
                                                            TOXICITY
                                                NO
                                                      CONVERT DATA TO
                                                      RANKITS OR RANKS
                                                NO
                                                                            T
                                                        ©
                                            1
                                                                    ONE-TAILED T-TEST FOR
                                                                    UNEQUAL VARIANCES ON
                                                                      RANKITS OR RANKS
ONE-TAILED T-TEST FOR
 UNEQUAL VARIANCES
                                  NO
                                                     DREDGED MATERIAL IS
                                                     NOT PREDICTED TO BE
                                                         ACUTELY TOXIC
  Figure D-l.     Water Column Toxicity Test Decision Tree.
                                                                              DRAFT

-------
                                               D-18
Tests of Assumptions
Following arcsine-transformation, the data were tested for normality (5) to determine whether parametric or
nonparametric procedures should be used. Table D-4 provides the results of tests for normality and equality
of variances for the example data. The value of Shapiro-Wilk's Wioi the arcsine-transformed data was 0.846,
with associated probability (P) = 0.051. Because this value of P exceeds 0.05 (o level from Table D-2, N=10,
balanced design), we conclude that the data do not depart significantly from the normal distribution (5), and
we now examine the results of the tests for equality of variances (6).

Bartlett's Test (from SYSTAT) and F' both indicated that the variances of arcsine-transformed data were not
significantly different for the  two treatments, with P>0.10 (a level from Table D-2, n=5, balanced design).
Thus, on the basis of these tests, we would proceed with a /-test for equal variances (7).

Two-sample t-tests
Table D-4 provides the results off-tests for equal (7) and unequal variances (8). The Mest for equal variances
indicated that survival in the 100% elutriate was significantly (P<0.05) less than in the dilution water (9). If
the data had been normally distributed with unequal variances, the Mest for unequal variances would have
been used. With the example data, both test results are the same, but this will not always be the case.
Table D-4.     Tests of Assumptions and Hypothesis Tests on Arcsine-Transformed Water Column Toxicity
               Test Example Data.
Null Hypothesis: Mean 100% Elutriate
Survival Equals Mean Dilution Water Survival3
Test
Normality Assumption:
Shapiro- Wilk's Test
Equality of Variances Assumption:
Bartlett's Test
FTest
Null Hypothesis:
f-Test (equal variances)
/-Test (unequal variances)
Mest on rankits (unequal
variances)
Test
Statistic
W=O.S46
F=0.5
F'=2.18
t= 12.734
t= 12.734
t= 4.631
Probability
P
0.051
0.47
0.468
<0.0001
<0.0001
0.0010
a
0.05
0.25
0.25
0.05
0.05
0.05
Conclusion
do not reject
do not reject
do not reject
reject
reject
reject
    Based on tests of assumptions, appropriate statistical test of null hypothesis is underlined.  Other test
    results are included for illustration only.
                                                                                     DRAFT

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	D-19	

Nonparametric Test

Nonparametric tests would generally not be performed on these data because the data did not depart signifi-
cantly from a normal distribution. However, the data were convened to rankits (10), and a Mest for unequal
variances (11) was conducted on the rankits (SAS Program WATTOX) for illustrative purposes.  The Mest
indicated that median survival in the 100% elutriate was significantly lower than in the dilution water (Table
D-4).

Statistical Power

The difference in survival between the 100% elutriate and the dilution water was so large (63%) that it was
easily detected (declared significant) even though there were only five replicates per treatment.  The power
of a Mest to detect such a large decrease in survival (d=O.S48 on the arcsine scale) when n=5 and 5=0.1055
(also on the arcsine scale) is >0.99.  However, it is reasonable to ask if n=5 is adequate for detecting smaller
differences.   For example, what sample size would be  required  to provide a iO.95  chance  (l-p=0.95;
2i.0=1.645) of detecting a reduction of survival to <;80%, with a=0.05 (z1.11=1.645)? In the example data, mean
arcsine-transformed dilution  water  survival was  1.4806 (=>99%  survival; back-transformation of means of
transformed values will not be the same as means based on original data, although the difference is trivial in
this case); the arcsine-transformed value for 80% survival is  1.1071, giving a reduction (d) of 0.3736 on the
arcsine scale; and the pooled s was 0.1055.  Using Eq. 9:


                    n = 2(1.645 +  1.645)2 (0.10552/0.37362)  + 0.25(1.6452)  =  2.40

Rounding up gives n=3. A more exact iterative computer program (SYSTAT DESIGN) based  on r-values
(Eq. 8) also yields «=3.  The sample size required for a 0.95  probability of detecting a reduction in survival
to 90% is n=6, again calculated with the  iterative program.  The minimum significant difference (i.e., the
difference we have a 0.50 probability of detecting) when n=5 is fa95 8(2s2/«)* or 1.86[2(0.10552/5)]* = 0.1241.
Subtracting that from the mean transformed dilution water survival, and back-transforming gives  95.5% sur-
vival. In other words, given the example data, the test can be expected to detect a reduction in survival from
=<99% to "95-96% approximately half the time.

When dilution water survival is near 100% and variation among replicates is low, as with the example data,
a test with «=5 replicates may be too powerful.  In many cases, we would declare survival of *90% in the
100% elutriate significantly lower than in the dilution water, yet  that *90% survival would be acceptable for
the dilution water.  For this reason, if survival in the 100% elutriate is not at least 10% lower than in the
dilution water, the difference  should not be considered significant and no statistical tests need be  performed.
It is important to remember that a statistically significant difference is not necessarily biologically significant (and
vice versa). If dilution water survival were lower, say 90% instead of 98%, and s remained the same, the Mest
would have less power. For example, n=13 would be required to provide a 0.95 probability of detecting a re-
duction in survival in the 100% elutriate to 80%. Much higher standard deviations can also be expected in
many toxicity tests.
                                                                                     DRAFT

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	D-20	

The SAS program WATTOX (Section D4.1) provides minimum significant difference and power of a /-test.
Power is determined for 10, 20, 30, 40 and 50 percent reductions in true population survival from the mean
dilution water survival.
D2.1.2     Calculating Median Lethal Concentration

In Tier III water column toxicity tests, the median lethal concentration (LCM) or median effective concentra-
tion (EC,,) are calculated when 100% elutriate survival is significantly lower than dilution water survival. The
LCjo is the concentration lethal to 50% of the test organisms; the ECM is the concentration causing some
sublethal effect (e.g., abnormality, immobility) in 50% of the test organisms. The remainder of this section
will discuss the LCM but all comments apply equally to EC50.  Steps and decisions in the LC^ determination
are shown in the decision tree in Figure D-2.  Numbers in parentheses in the text refer to numbered nodes
of the decision  tree.

Ideally, data for at least five elutriate concentrations should be available to calculate an LC50, although most
methods described below can be used for fewer concentrations. The control or dilution water survival is not
included.  Survival in the lowest elutriate concentration must be at least 50% (1); otherwise the test must be
repeated using  lower concentrations (2).  An LC50 should not be calculated unless at least 50% of the test
organisms die in at least one of the serial dilutions (3). If there are no mortalities greater than 50%, then the
LC50 is assumed to be *100% (4).

If the conditions in  (1) and (3) are met,  then replicate mortality data for each concentration are pooled (5)
for calculation of LC50 (6).  The Probit method (7) can be used if the data meet the requirements of the Probit
method listed below and fit the probit model  (8).  The Trimmed Spearman-Karber (TSK) and Logistic
methods (described below) are acceptable substitutes for the Probit method, provided  that the data meet the
requirements of these alternative methods. If the data do not meet the requirements of the Probit method
or alternatives, then the Linear Interpolation method should be used (9).  When an LC50 value  has been
determined, 1% of that value is entered into the mixing  model (10) provided in Appendix C  for mixing zone
evaluation.

Calculation of LC50 values is also recommended for reference toxicant tests to determine the relative health
of the organisms used in toxicity and bioaccumulation testing (Section 13.3.17.2).
D2.1.2.1   Methods For Calculating LCM

Stephan (1977) and Gelber et al. (1985) provide careful reviews of LC50 estimation procedures. In addition,
USEPA (1985) discusses in detail the mechanics of calculating LC50 using current methods and contains, as
an appendix, computer programs for each statistical method.  The most commonly used methods are the
                                                                                    DRAFT

-------
                                                 D-21
                        SURVIVAL DATA -
               ELUTRIATE CONCENTRATION SERIES
©
             IS SURVIVAL IN LOWEST ELUTRIATE
                 CONCENTRATION  a  50%?
  REPEAT TEST USING
LOWER CONCENTRATIONS
             IS SURVIVAL IN AT LEAST ONE ELUTRIATE
                    CONCENTRATION  * 50%?
                                                         NO
                                                                         LCSO > 100% vol./vol.
                                                                    (ASSUME LCSO  3t 100% vol./vol.
                                                                             ELUTRIATE)
                    ENTER POOLED DATA FOR
                     EACH CONCENTRATION
                      RUN LCSO PROGRAM
                 IS PROGRAM ABLE TO CALCULATE
                  LCSO USING PROBIT METHOD? *
                                                         NO
                           YES
                            T
               IS THE PROBABILITY OF GOODNESS OF
                           FIT a 0.05?
                                                         NO
                           YES
                            I
                 USE LCSO  FROM PROBIT METHOD
                                                                USE LCSO FROM LINEAR
                                                                INTERPOLATION METHOD
                                 COMPARE 0.01 LCSO TO PREDICTED CONCENTRATION
                                        BASED ON MIXING MODEL (APPENDIX C)
         Trimmed Spearman-Karber and logistic methods are acceptable substitutes for Probit method
Figure D-2.
                      Decision Tree.
                                                                                     DRAFT

-------
	D-22	

Probit, Trimmed Spearman-Karber (TSK) and Linear Interpolation. This Appendix recommends use of the
Probit, TSK or Logistic methods if the data are appropriate; otherwise the Linear Interpolation method may
be used (Figure D-2). In general, results from different methods should be similar. Programs commonly used
to calculate LC^ are PROBIT, developed for and available from the USEPA (Environmental Monitoring and
Support Laboratory, Cincinnati, OH), and several programs developed by Dr. C.E. Stephan of the USEPA
Environmental Research Laboratory in Duluth, Minnesota.  Procedures  in statistical packages such as SAS
or SYSTAT may not be easily adaptable for routine catenations of LCj^s, and specialized  packages are
generally preferred. This Appendix does not include SAS programs for LC^.

Probit

The Probit method is based on regression of the probit of mortality on the log of concentration.  A probit is
the same as  a z-score; for example, the Probit corresponding to 70% mortality is za70 or »0.52. The LC^ is
calculated from the regression, and is the concentration associated with z=0 (mortality = 50%).  The Probit
method can  be used whenever the following conditions are met:
    •   there are at least two concentrations with partial mortality (i.e., >0 and <100%)

    •   the data points fit the probit regression line reasonably well.


The first condition is necessary because the regression line is estimated from the partial mortalities.  The
second condition, called goodness-of-fit, can be tested by the x2 statistic, which is a measure of the distance
of the data points from the regression line.  A low x2 indicates a good fit. By convention, the fit is considered
adequate if the P-value for %2 is >0.05 (in other words, goodness-of-fit is rejected if PsO.05). Programs such
as PROBIT will only provide %2, in which case %2 should be compared against tabled values with k - 2 df, where
k is the number of partial mortalities. If there are only two partial mortalities (k=2), then there are 0 df, and
the goodness-of-fit cannot be tested (i.e., a line between two points  is always a perfect fit). When there are
only two partial mortalities, the LCj,, is identical to the LC50 which would be calculated by Linear Interpolation
(see below) with mortality expressed on a probit scale. Goodness-of-fit can also be assessed by eye, if the data
are plotted on log-probit paper, or if the computer program provides a plot.

Linear Interpolation Method

The Linear Interpolation method should be used when:

    •   there are 0 or 1 partial mortalities

    •   the data do not fit the Probit (or Logistic) models
                                                                                    DRAFT

-------
	D-23	

The Linear Interpolation method should also be used when LCsos are calculated and compared over an
extended time series (i.e., for tracking reference toxicant results), because inevitably, one or more data sets will
fail to meet the requirements for the Probit, TSK or Logistic methods. Linear Interpolation may also be used
if programs for  the  other  methods are unavailable,  but we strongly recommend that investigators have
programs available for one  or more of the other methods.

The Linear Interpolation method calculates an LCSO by interpolation between the two concentrations with
mortality nearest to, and on either side of 50%.  The interpolation is made on a log concentration scale, using
the following formula:
                                                                                          (Eq.H)
                                                   My ~
where   CL = concentration with mortality nearest to and below 50%
        Cu = concentration with mortality nearest to and above 50%
        ML = % mortality at CL
        MU = % mortality at Cu.

If there are no partial mortalities, the formula simplifies to:
For the example data given in Table D-3, CL=25% elutriate (log= 1.398); ML=28% mortality; Cu=50% elu-
triate (log= 1.699); and Mv=54% mortality. Therefore:

                                      5°  " 28) (L699) * (54 " S0) (1 398)
                          50                        54-28

or 44.9%.

The formula and example given above express  mortality on an arithmetic (untransformed) scale.  Some
computer programs or investigators may use arcsine-transformed mortalities (Stephan, 1977; see Section
D.2.1.1.1 Tests of Assumptions'). One could also express mortality on a probit or logit scale, if there were one
partial mortality on each side of 50%. In those cases, the Linear Interpolation should produce the same LC^
estimate as the Probit or Logistic methods. In this manual, we recommend the use of untransformed mortality
for simplicity and consistency.  However, LCSO estimates using other scales can easily be calculated for
comparison.
                                                                                   DRAFT

-------
	D-24	

Trimmed Spcarman-Karber (TSK.) Method

The TSK method is a nonparametric method that can be calculated by hand using the procedure in Gelber
et al. (1985).  The calculations can be tedious, especially for processing large numbers of tests, and computer
programs are usually used. The method is labelled "trimmed" because extreme values (mortality much higher
or lower than 50%) are "trimmed" or removed prior to calculation of the LC50.  Thus, the LQ,, is calculated
using points near 50% mortality, which may produce a more robust estimate.  The TSK method can be used
in many cases where the Probit method is unsuitable.  Access to appropriate computer programs, and
difficulties in deciding what values to trim are probably the major factors limiting widespread use of the TSK
method. Investigators with access to reliable programs should not hesitate to use the TSK method whenever
there are two or more partial mortalities. Information concerning TSK computer programs may be obtained
from the USEPA Environmental Research Laboratories  in Athens, GA, or Duluth, MN, or CSC/USEPA,
Cincinnati, OH.

Logistic Method

The Logistic method is similar to the Probit method except that mortalities are converted to logits rather than
probits. A logit is log [Af/(100 - M)], where M is % mortality. The LC50 is derived from a regression of logits
on log concentration.  As with the Probit method, the Logistic method can be used whenever there are two
or more partial mortalities, and the data fit the regression line.  Logistic regression is not commonly used in
aquatic toxicology only because  Probit  programs are more available, but the two methods  are equally
acceptable.  Logistic  regression programs in SAS and SYSTAT are designed for  complex analyses and
comparisons of logistic regressions, and may be inconvenient to use for simple and routine  calculations of LC^
for single tests.
D2.1.2.2    Analysis of Example Data

Table D-5 provides LC^ estimates calculated by several different methods using the example data in Table D-3.
In all cases, the data from the five replicates for each concentration were pooled, and entered as the number
responding (dying) out of 100. Because pooling over replicates ignores any additional variance in survival among
replicates (Le., beyond the expected error from sampling the binomial distribution), the confidence limits provided
by the programs may not be accurate and should not be reported or used.  Because the LCSO is required only for
use in the mixing model (Appendix C), confidence limits are not needed.
The Probit LCj,, was calculated with the EPA PROBIT program, and was almost identical to the Logistic
calculated using the SYSTAT LOGISTIC program.  The %2 goodness-of-fit for the Probit line was 1.756,
indicating a good fit (P>0.05 with 4-2 = 2 df), which could be verified by examining the plot provided (Fig-
ure D-3).  The LC50 estimated by Linear Interpolation, with untransformed mortality, was almost identical to
                                                                                   DRAFT

-------
                                               D-25

Table D-5. Calculated LC50 Values for Example Water Column Toxicity Test Data.
Method
Probit
Linear Interpolation
- untransformed mortality
- arcsine-transformed mortality
Trimmed Spearman-Karber
Logistic
LCj, Estimate
(% v/v)
52.6
44.9
45.1
48.4
52.6
the LC50 calculated using arcsine-transformed mortality.  The TSK LCM was calculated using a program
modified from an original program described in Hamilton et al. (1977), and was intermediate between the
Linear Interpolation and regression (Probit and Logistic) estimates.

The various estimates in Table D-5 differed by up to 7.7% elutriate, which is not unusual or alarming. The
Probit or Logistic LC50 would be the preferred estimate, because the regression  lines fit the data well, and the
regression methods use more of the data in such cases.  However, any of the estimates would be adequate for
use in the mixing model in Appendix C, because the imprecision and uncertainty involved in the model
calculations and estimates are undoubtedly far greater than the differences among the LCSO estimates.
D2.2       Tier III Benthic Toxicity Tests

The objective of Tier III benthic toxicity tests is to determine if sediments taken from a potential dredge site
are significantly more toxic than a reference sediment.  The test procedure is described in Section 11.2. The
statistical analysis recommended below assumes that individual dredge sites are relatively large, and that a
decision about potential sediment toxicity, and subsequently about disposal options, will be made indepen-
dently for each site.  If only one dredge site is tested, and compared to a reference sediment, statistical analysis
is the same as that given in Section D2.1.1 for comparison of 100% elutriate and dilution water (Figure D-l
and SAS program WATTOX in Section D4.1). However, in many cases, more than one dredge site is tested
simultaneously with one reference sediment. In those cases, recommended statistical methods will differ from
the two-sample case. Methods for comparison of more than one dredged sediment with a reference sediment
are described below, and computer procedures are given  in SAS program BENTOX (Section D4.2).
D2.2.1     Methods

Fisher's Least Significant Difference (LSD)

Fisher's Least Significant Difference (LSD) is the appropriate parametric statistical test for assessing differ


                                                                                    DRAFT

-------
                                               D-26
Probit
   10+
    9+
    8+
    7 +
    6+
                                              . . .o
      -                                   o . . . .
    5+
      -                          . .o
      ~                       • * • •
    4+                    .O..




      ~~        • •
    3 +





    2+





    1+
     0+
       _+	+	h	+	+	+	1—
       EC01            EC10     EC25      EC50       EC75     EC90            EC99
      Figure D-3.     Probit Plot of Water Column Toricity Test Example Data.
                                                                                DRAFT

-------
_ D-27 _

ences in survival or other response when more than two means are being compared. This a posteriori multiple
comparison technique is discussed in many statistical texts, e.g., Steel and Torrie (1980); SAS Institute, Inc.
(1988b); Snedecor and Cochran (1989); and Wilkinson (1990).  The LSD controls the pairwise Type I error
rate rather than the experimentwise Type I  error rate.  This  means that  the  Type I error rate for each
comparison is held to the preset a even though the overall Type I error rate for all comparisons (i.e.,
experimentwise error rate) may be higher.  A test that controls the pairwise error rate is appropriate because
disposal decisions  are to be made independently for each dredge site regardless of how many sites are
compared to the same reference. The LSD replaces the previously recommended Dunnett's test, which is not
appropriate because it controls experimentwise error rate.

The LSD is usually performed in conjunction with analysis of variance (ANOVA), and only if the data meet
the ANOVA assumptions of normality and equal variances. The ANOVA is conducted primarily to provide
the mean square error (MSE), which is an estimate of the pooled variance across all treatments. The ANOVA
F-statistic and its associated probability are ignored in this application.

The test statistic for the LSD is t, calculated in much the same way as for a Mest:


                                 t-Gt-xjl JMSE (1/n,  + l//ij)                        (E* 13)


This ^-statistic is compared against the distribution of Student's t with N - k degrees of freedom, where N is
the total number of observations (Sn) and k is the number of treatments including the reference. A r-statistic
is computed for each possible pair of treatments in the analysis.

The MSE can be calculated as:
                                MSE = S   (nt - 1)] / S(n. - 1)  ,

where s? and n,  are the variance and number of replicates for the ah treatment.  The term S(n( - 1) is
equivalent to N - k.

If sample sizes are equal, then:


                                  MSE (1/n, + I/BJ)  = 2MSE/n  .                          (Eq. 15)
The major advantage of using the LSD as opposed to conducting individual two-sample Mests comparing each
dredged sediment to the reference is that the MSE is a better estimate of the true population variance than
the pooled variance calculated from only two samples.  Consequently, the LSD test is more powerful, as
                                                                                  DRAFT

-------
	D-28	

reflected in the greater df for the calculated t. It also follows that a pooled variance should only be calculated,
and the LSD test conducted, if the variances for the treatments are not significantly different.

Tests of Assumptions

The Shapiro-Wilk's Test described in Section D2.1.1.1 can be used to test for normality when more than two
treatments are compared.  If the data are not normally distributed, even after an appropriate transformation,
then nonparametric tests should be used (see Nonparametric Tests  below).

Bartlett's Test, Levene's Test, F^ or Cochran's Test can be used to test for equality of variances. If there
are more than two samples, then F^ is equal to the largest variance divided by the smallest variance.  If
variances are significantly unequal, even after transformation, then each dredged sediment should be compared
with the reference using two-sample r-tests.

Nonparametric Tests

When parametric tests are not appropriate for multiple comparisons because the normality assumption is
violated, the data should be converted to rankits, and the rankits should be tested for normality and equality
of variances.  If these assumptions are not violated, an LSD is then performed on the rankits (Conover, 1980,
refers to this as van der Waerden's Test).  Tests  performed on rankits are robust to departures from normality,
and can still be used when the normality assumption is  violated. Rankits will rarely fail tests for normality,
partly because a normal distribution is imposed over the entire data  set. The rankit data may fail the test for
equality of variances, but then Mests can be conducted for each pair of treatments to be compared. If rankit-
transformed data fail normality tests, it is probably safest to use the /-tests for unequal variances, as some tests
for equality of variance are not robust when data are non-normal.

When rankits cannot be easily calculated, the  original  data may be converted to ranks (using SAS PROC
RANK, for example). Equality of variances should be tested after the data are ranked. There is a common
misconception that nonparametric tests can be used when variances are not equal as well as when data are not
normally distributed.  However, nonparametric tests are not very robust if the variances of the ranks are not
similar among treatments. Bartlett's Test should not be used to test equality of variances of ranks, as ranks
will  follow a uniform, rather  than a normal  distribution, and Bartlett's Test  is unduly sensitive to non-
normality.  Other tests discussed in Section D2.1.1.1 Tests for Equality of Variances  may be used on ranks;
there are also nonparametric tests for equality of variances provided in Conover (1980).

If the variances of the ranks are not significantly different, the Conover T-Test (Conover, 1980)  should be
performed.  This test can most easily be conducted by performing an LSD on the ranks.  If the variances of
ranks are significantly unequal, a one-tailed r-test for unequal variances should be performed (using ranks) for
each pair of treatments to be compared.
                                                                                     DRAFT

-------
                                               D-29

Statistical Power

Power calculations for the LSD are the same as for the r-test (see Eq. 8), except that the degrees of freedom
for fj.. and t^ are N - k, and MSB replaces s2 if variances are equal (and an estimate ofMSE is available):
                                     ' 2
If the z-approximation (Eq. 9 with MSE replacing s2) is used to calculate samples size, the result will be a
slight overestimate, although the overestimation is  rarely of practical importance.  Finally,  the minimum
significant difference should be reported for LSD tests.  Note that the test is named the Least Significant
Difference because another way to conduct the test is to compare the observed differences to the minimum
significant difference.

If an increase in power (1-P) is desired, because variance is high or sample size low, one effective method of
increasing power is to increase the number of reference replicates rather than increase the sample size for each
treatment.  It is even possible to increase power without increasing overall sample size by increasing sample
size for the reference, and decreasing sample size for the dredged sediments. The optimal apportionment of
replicates is to make  the sample size for  the reference fk times  the sample size for  the other sediments
(Dunnett, 1955).   Increasing sample size  for the reference  sediment is effective because the reference is
involved  in every comparison, whereas the dredged sediments are involved in only one comparison each.
D2.2.2     Analyses of Example Data

Table D-6 presents survival data from a hypothetical benthic toxicity test comparing survival from  three
dredged sediments with reference sediment survival.  The example data are used to illustrate the steps in
benthic toxicity data analysis, with numbers in parentheses in the text referring to numbered nodes in the
decision tree (Figures D-4A.B). In this example, survival in the control (data not shown) was *90%, indicat-
ing the acceptability of the test (Figure D-4AJ). Mean survival in all dredged sediments was more than 10%
below mean survival in the reference sediment, indicating  that the significance of the reductions should be
tested statistically (2). All data were arcsine-transformed prior to analyses (3). Data were analyzed using SAS
program BENTOX (Section D4.2), and results for the analyses are given in Section D4.2.2.
                                                                                    DRAFT

-------
                                               D-30
Table D-6. Number of Survivors in a Hypothetical Benthic Toxicity Test.
Replicate*
1
2
3
4
5
Total
Mean
SE
Treatment
Reference
20
20
19
19
20
98
19.6 (98%)
0.24
Sediment 1
17
16
18
17
15
83
16.6 (83%)
0.51
Sediment 2
15
16
13
17
11
72
14.4 (72%)
1.08
Sediment 3
17
12
10
16
13
68
13.6 (68%)
1.29
'   20 organisms per replicate at initiation of test

Tests of Assumptions

Following arcsine-transformation, the data were tested for normality (4) to determine whether parametric
(Figure D-4A) or nonparametric (Figure D-4B) procedures should be used.  Results of tests for normality (4)
and equality of variances (5) are provided in Table D-7. The P-value for the Shapiro-Wilk's Test was 0.32,
indicating no significant departure from normality because P exceeds 0.01 (a level in Table D-2 for N=20,
balanced data).  Bartlett's Test, Levene's Test, and Fma[ all indicated that variances were  not  significantly
different among groups, as all P-values were >0.10 (a level in Table D-2 for n=5, balanced data). Note that
these three tests were included for the sake of comparison, but generally only one of them would be
conducted.  Because the data are normally distributed and variances are not significantly different,  the LSD
is the most appropriate test for comparing each dredged sediment to the reference (6).

Parametric Tests

Relevant results from the LSD test are provided in Table D-7 (note that LSD results are given separately for
each dredged sediment-reference sediment comparison, but only one LSD test is actually performed, comparing
each pair of sediments simultaneously).   The P-values for the LSD comparisons of each sediment with the
reference were all much less than 0.05; thus, we conclude that survival in each of the dredged sediments was
significantly less than reference sediment survival (7). SAS output for the LSD test (Section  D4.2.2) does not
provide f-values and probabilities for the individual comparisons, and it is not necessary to calculate these.
                                                                                     DRAFT

-------
                                           D-31
                 SURVIVAL DATA -
         DREDGED SEDIMENTS VS. REFERENCE
                CALCULATE MEANS
                CONTROL SURVIVAL
                   STANDARD?
                                                       REPEAT TEST
            IS SURVIVAL IN AT LEAST ONE
      DREDGED SEDIMENT AT LEAST 10% LOWER
           THAN SURVIVAL IN REFERENCE?
                                               NO DREDGED SEDIMENTS SIGNIFICANTLY
                                                   MORE TOXIC THAN REFERENCE
               ARCSINE TRANSFORM
                                                                     USE NON-PARAMETRIC
                                                                     PROCEDURES (FIG. 4B)
ARE DATA NORMALLY DISTRIBUTED?
                                                     ONE-TAILED T-TESTS FOR EACH
                                                   DREDGED SEDIMENT < REFERENCE
     ARE VARIANCES EQUAL?
               ONE-TAILED LSD TEST
          COMPARING DREDGED SEDIMENTS
                 WITH REFERENCE
                                                             DREDGED SEDIMENT NOT
                                                            SIGNIFICANTLY MORE TOXIC
                                                                THAN REFERENCE
IS SURVIVAL IN DREDGED SEDIMENT
 SIGNIFICANTLY < SURVIVAL IN THE
     REFERENCE SEDIMENT?
         DREDGED SEDIMENT SIGNIFICANTLY
           MORE TOXIC THAN REFERENCE
Figure D-4A.   Benthic Toxicity Test Decision Tree (Parametric Tests).
                                                                            DRAFT

-------
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                                               D-33
Table D-7.     Tests of Assumptions and Parametric Tests of Hypotheses on Arcsine-Transformed Benthic
               Toxitity Test Example Data.
Null Hypothesis: Mean Dredged Sediment Survival Equals Mean Reference Sediment Survival*
Test
Normality Assumption:
Shapiro- Wilk's Test
Equality of Variances Assumption:
Bartlett's Test
Levene's Test
F^Test
Null Hypotheses:
Sediment 1 = Reference
LSD Test
f-Test (unequal variances)
Sediment 2 = Reference
LSD Test
f-Test (unequal variances)
Sediment 3 = Reference
LSD Test
f-Test (unequal variances)
Test
Statistic
W=0.946
f=0.6
F=1.74
F =44
-* max ^'^
t=4.n
f=5.09
f=5.73
f=5.63
t=6.25
t=5.57
Probability
P
0.322
0.61
0.199
>0.25
0.0017
0.0009
0.0002
0.0003
0.0001
0.0004
a
0.01
0.10
0.10
0.10
0.05
0.05
0.05
0.05
0.05
0.05
Conclusion
do not reject
do not reject
do not reject
do not reject
reject
reject
reject
reject
reject
reject
    Based on tests of assumptions, appropriate statistical tests of null hypotheses are underlined.  Other test
     results are included for illustration only.
SAS indicates significant differences by using different letters under the T Grouping" column. Mean reference
survival was highest (A); mean survivals for sediments 1 (B) and 2 (BC) were significantly less than reference
but not different from each other, and sediment 3 mean survival (C) was significantly lower than reference and
sediment 1 but not sediment 2.

If the variances had been unequal, survival data would have been compared using Mests (8). These results
are included in Table D-7 for illustration. Again, the P-values indicate that all dredged sediment survivals
were significantly less than reference sediment survival. Note that these P-values are one-half those given in
the output from  SAS program BENTOX in Section D4.2.2, because the SAS TTEST procedure returns two-
tailed, rather than  one-tailed probabilities.

Nonparametric Tests
Although the arcsine-transformed example data did not violate parametric hypothesis testing assumptions,
nonparametric tests were performed to illustrate the steps in the nonparametric decision tree (Figure D-4B).
The  example data were converted using both rankits (J) and  ranks  (2), and  the  appropriate  tests of
assumptions were conducted (Table D-8).  The rankits passed both the normality (3) and equality of variances
                                                                                    DRAFT

-------
                                               D-34
(4) tests, so the next step would be the LSD on rankits (5). Had either of these assumptions been violated,
Mests for unequal variances would have been performed on the rankits (6).  If the ranks had failed the
Levene's Test for equality of variances (7), /-tests for unequal variances would have been performed on the
ranks (8), rather than the Conover J-Test (9). Results for all of these nonparametric hypothesis tests are
shown in Table D-8. SAS Program BENTOX does not perform Levene's Test on ranks, the Conover 7-Test,
or 2-sample Mests on  ranks, as SAS can easily calculate rankits, and ranks-based tests would not be needed.
The P-values for the nonparametric hypothesis tests in Table D-8 were in most cases slightly greater than those
for the parametric tests, suggesting slightly lower power for the nonparametric tests. Nevertheless, all tests
indicated that survival was significantly reduced in the dredged sediments compared to reference sediment
survival. These results could easily have been predicted prior to analyses, because survival in the  dredged
sediment samples did  not overlap with survival in the reference sediment samples (Table D-6).
Table D-8.     Tests of Assumptions and Nonparametric Hypothesis Tests on Benthic Toxicity Test Example
               Data Converted to Rankits and Ranks.
Null Hypothesis:
Median Dredged Sediment Survival Equals Median Reference Sediment Survival

Test
Normality Assumption:
Shapiro- Wilk's Test (rankits)
Equality of Variances Assumption:
Levene's Test (rankits)
Levene's Test (ranks)
Null Hypotheses:
Sediment 1 = Reference
LSD Test (rankits)
/-Test (rankits, unequal variances)
Conover J-Test (ranks)
/-Test (ranks, unequal variances)
Sediment 2 = Reference
LSD Test (rankits)
/-Test (rankits, unequal variances)
Conover T-Test (ranks)
/-Test (ranks, unequal variances)
Sediment 3 = Reference
LSD Test (rankits)
/-Test (rankits, unequal variances)
Conover 7-Test (ranks)
/-Test (ranks, unequal variances)
Test
Statistic

W=0.982

F=1.18
F=2.25


/=3.05
/=4.57
/=3.04
/=4.27

/=4.71
/=5.44
/=4.90
/=5.80

/=5.28
/=4.91
/=5.30
/=5.51
Probability
P

0.940

0.349
0.122


0.0079
0.0011
0.0080
0.0036

0.0008
0.0007
0.0006
0.0012

0.0004
0.0019
0.0004
0.0018

a

0.01

0.10
0.10


0.05
0.05
0.05
0.05

0.05
0.05
0.05
0.05

0.05
0.05
0.05
0.05

Conclusion

do not reject

do not reject
do not reject


reject
reject
reject
reject

reject
reject
reject
reject

reject
reject
reject
reject
                                                                                    DRAFT

-------
                                               D-35

Statistical Power

From Eq. 11, the minimum significant difference (dmin, when ^=0) for the parametric LSD test was:
= 1.746[2(0.01618)/5]* = 0.1405, where v = 16 df.  Subtracting 0.1405 from the mean arcsine-transformed
survival in the reference (1.481), and back-transforming gives 95%. That is, any survival less than 95% mea-
sured in a sample would be significantly lower than in the reference, and we would have a 0.50 probability of
detecting a reduction in survival in any case where true population survival was 95%. Modifying Eq. 10, the
probability (power or 1-0) of detecting a difference if true population survival in a dredged sediment is <90%
can be determined by:
                                                                                          (Eq. 18)
= (1.481 - 1.249)/[5/2(0.01618)f - 1.746 = 1.138.  Using the SAS PROBT function to determine 1-p for
t = 1.138 with 16 df, power = 0.86. As with the water column toxicity test example data, the level of replication
for the benthic toxicity example data is adequate to detect any reductions in survival that would be considered
biologically significant.   Investigators can expect lower reference survival  and/or greater variance, and
consequently less power, in real toxicity tests.

Suppose that we required an increase in power, but could not afford to add any more replicates. The optimal
solution, assuming that variance could not be reduced by improving laboratory practices, would be to use 8
replicates for the reference, and 4 for each of the dredged sediments.  The overall sample size remains 20.
Note that a ratio of reference:dredged sediment replicates of 8:4 (2:1) is approximately equal to the optimal
ratio of V"k:l or 1.73:1 (Jt=3 with 3 dredged sediments). Assuming thatMS£=0.01618, as above, the minimum
significant difference for an LSD test,  again with  16 df, would be:
                                       = ( 'i-..v   AfSEn  +  I*                             q- 19)
= 1.746[0.01618(l/4 + 1/8)]* = 0.1360. This value is lower, although by <5%, than the minimum significant
difference of 0.1405 for equal  sample sizes of 5.  The increase in power using the optimal ratio of refer-
ence:dredged sediment replicates will be greater when k is greater (more sediments tested).

SAS program BENTOX (Section D4.2) provides power calculations for the LSD test when true population
survival from a dredged sediment is 10, 20, 30, 40 and 50 percent lower than mean reference sediment survival.
                                                                                    DRAFT

-------
	D-36	

D3.0           BIOACCUMULATION

Bioaccumulation tests described in Section 12 are applied to determine whether an organism's exposure to the
dredged material is likely to cause an elevation of contaminants in its tissues, i.e., bioaccumulation. Bioac-
cumulation tests may be conducted in the laboratory or in the Meld. Data analysis for these tests uses statisti-
cal procedures that have already been described  for benthic toxicity test data analysis. These procedures are
illustrated with example data in the  following sections.

Bioaccumulation tests are especially prone to the departures from ideal test conditions discussed in Section
Dl.O. A particularly vexing problem is the statistical treatment of contaminant concentrations reported only
as less than some detection limit. Such non-numeric data cannot be statistically analyzed using procedures
in this Appendix unless numeric values are substituted for the less-than detection limit observations.  Tech-
niques for handling less-than data will be evaluated by the USACOE, and recommendations will be published
in an Applications Guide as a supplement to this Appendix.  Until such recommendations are promulgated,
we suggest substituting one-half the detection limit for each less-than observation.  This is an interim recom-
mendation only, for the sake of consistency, and does not necessarily represent the optimum technique for treatment
of less-than data.
D3.1           Tier III Single-Time Point Laboratory Bioaccumulation Study

The Tier III single-time point laboratory bioaccumulation test produces tissue concentration measurements
for each contaminant of concern. Table D-9 presents example results for one contaminant from a hypothetical
laboratory test. Chemical analysis of the tissue samples from each replicate shows that concentrations of the
example contaminant varied among and within sediments. Two types of analyses may be performed on these
data:

        •      comparisons between each dredged sediment and the reference, and

        •      comparisons with an action level when applicable.
Although Section 6.3 stipulates that applicable comparisons with an action level be conducted first, the statis-
tical analysis can be performed more efficiently if comparisons with the reference are done first.  Computer
procedures  for statistical analysis of single-time point bioaccumulation data  are given  in SAS program
BIOACC (Section D4.3).
                                                                                     DRAFT

-------
                                               D-37
Table D-9.     Results from a Hypothetical Single-Time Point Bioaccumulation Test, Showing Contaminant
               Concentrations (u.g/g) in Tissues of Animals Exposed to Different Treatments.
Replicate
1
2
3
4
5
Mean
SE
Treatment
Reference
0.06
0.05
0.05
0.08
0.09
0.066
0.008
Sediment 1
0.16
0.19
0.18
0.22
0.31
0.212
0.026
Sediment 2
0.24
0.10
0.13
0.18
0.30
0.190
0.036
Sediments
0.13
0.05
0.17
0.08
0.22
0.130
0.030
D3.1.1
Comparisons with a Reference Sediment
Analysis of the example data follows the decision tree steps in Figures D-5A and 5B, with numbers in paren-
theses in the text referring to numbered nodes of the decision trees. The objective of this type of analysis is
to determine whether organisms exposed to the dredged sediments accumulate greater tissue contaminant
levels than organisms exposed to the reference sediment.  One-sided tests are appropriate because there is
little concern if bioaccumulation from a dredged sediment is less than bioaccumulation from the reference
sediment. If mean tissue concentrations of contaminants of concern in organisms exposed to a dredged sedi-
ment are less than or equal to those of organisms exposed to the reference sediment (7), the dredged sediment
meets the guidelines (Section 6.3), and  no statistical analysis is required.

If only one dredged sediment is compared to the reference, then the procedures described in Section D2.1.1.1
(tests of assumptions followed by a Mest using a transformation or rankits if necessary)  for comparing two
samples are used. If more than one sediment is compared to the reference, then  the procedures described in
Section D2.2.1 (tests of assumptions followed by LSD, Mests, or nonparametric equivalents) are used. Because
contaminant concentration  data are not easily expressed as  proportions, the arcsine transformation is not
appropriate.  The  raw data are analyzed first and, if necessary, a logarithmic (either natural or  base  10)
transformation may be employed.  Although other transformations (such as square root)  are possible, we
recommend the  log transformation  because contaminant concentration data  often follow  a  lognormal
distribution.  As always, tests of assumptions must be rerun on the data following transformation. If the
transformed data violate the normality assumption, then data are converted to rankits (or ranks)  and  the
assumptions are retested.
                                                                                    DRAFT

-------

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	D-40	

The data in Table D-9 were analyzed using SAS program BIOACC (Section D4.3), and the results are reported
in Tables D-10 and D-ll.  The probability value for Shapiro-Wilk's Test (2) was >0.01 (a level in Table D-2
for N=20, balanced data), indicating no significant departure from normality.  If the raw data had failed the
normality test, then a log transformation (5) would be applied and the Shapiro-Wilk's Test rerun (2).  If the
log-transformed data still departed significantly from normality,  then nonparametric  hypothesis testing
procedures would be performed (Figure D-5B); these procedures are described in Section D2.2.1.

The P-value for Levene's Test (4) was >0.10 (a level in Table D-2, n=5, balanced data), indicating that
assumption of equality of variances need  not be rejected for the raw data.  If the variances had been
significantly unequal, a log transformation would have been applied (3) and the tests of assumptions (2,4)
rerun. Data that passed the normality test but failed the test for equality of variances would be analyzed using
a Mest for each dredged sediment-reference sediment  comparison (5).

Because the example data passed both tests of assumptions, the LSD (6) was conducted on the untransformed
data to compare bioaccumulation from each dredged sediment with bioaccumulation  from the reference
sediment. LSD results indicated that mean tissue levels for organisms exposed to dredged sediments  1 and
2 (but not 3) were significantly greater than mean tissue levels for organisms exposed to the reference sediment
(Table D-10).

For the sake of illustration, Table D-10 also includes results for log-transformed example data and for f-tests.
Table D-ll gives nonparametric test results for the example data. Note that the different statistical tests give
conflicting hypothesis test conclusions for the sediment 3-reference sediment comparison, because the P-values
of the tests are close to a.  This situation will often arise in the analysis of actual bioaccumulation data.  Once
again, it  is not acceptable to conduct several different statistical tests in order to choose the results one prefers.
For dredged sediment disposal evaluations, the decision  trees in this Appendix should  be followed to
determine the appropriate statistical procedures in any given situation.  In the case of the example data, the
tests of assumptions indicate that the appropriate hypothesis testing procedure is the LSD test using untrans-
formed data, and the results of this test should be accepted. However, in making decisions concerning  dis-
posal, it is entirely appropriate to consider that the significance of the sediment 3-reference sediment compari-
son is marginal. The power of the LSD test (calculated below) should also be taken into consideration.

Power calculations for the example data are performed on the untransformed data.  Using Eq. 17, the mini-
mum significant difference for the parametric LSD test was:

                              dmin =  1.746[2(0.003763)/5f = 0.0677 jig/g.

SAS conveniently provides this value as the "Least Significant Difference" in the GLM or ANOVA procedures
when the LSD test is requested (and sample sizes are equal).
                                                                                     DRAFT

-------
                                               D-41

Table D-10.    Tests of Assumptions and Parametric Hypothesis Tests on Untransformed and
               Logio-Transformed Bioaccumulation Example Data.
Null Hypothesis: Mean Dredged
Sediment Bioaccumulation Equals Mean Reference Sediment Bioaccumulation'
Test
Normality Assumption:
Shapiro-Wilk's Test
Untransformed data
Log-transformed data
Equality of Variances Assumption:
Levene's Test
Untransformed data
Log-transformed data
Null Hypotheses:
Sediment 1 = Reference
LSD Test
Untransformed data
Log-transformed data
r-Test (unequal variances)
Untransformed data
Log-transformed data
Sediment 2 = Reference
LSD Test
Untransformed data
Log-transformed data
f-Test (unequal variances)
Untransformed data
Log-transformed data
Sediment 3 = Reference
LSD Test
Untransformed data
Log-transformed data
/-Test (unequal variances)
Untransformed data
Log-transformed data
Test
Statistic
W=0.95S
W=0.980

F=2.15
F=2.19


t=3.76
f=4.45
7=5.30
7=7.04

7=3.20
7=3.84
7=3.33
7=4.34

7-1.65
t=2.20
7=2.03
7=1.98
Probability
P
0.511
0.921

0.134
0.129


0.0028
0.0011
0.0020
< 0.0001

0.0063
0.0025
0.0129
0.0020

0.0688
0.0295
0.0523
0.0495
a
0.01
0.01

0.10
0.10


0.05
0.05
0.05
0.05

0.05
0.05
0.05
0.05

0.05
0.05
0.05
0.05
Conclusion
do not reject
do not reject

do not reject
do not reject


reject
reject
reject
reject

reject
reject
reject
reject

do not reject
reject
do not reject
reject
    Based on tests of assumptions, appropriate statistical tests of null hypotheses are underlined. Other test
    results are included for illustration only.
                                                                                   DRAFT

-------
                                               D-42
Table D-ll.    Tests of Assumptions and Nonparametric Hypothesis Tests on Bioaccumulation Example
               Data Converted to Rankits and Ranks.
Null Hypothesis: Median
Dredged Sediment Bioaccumulation Equals Median Reference Sediment Bioaccumulation

Test
Normality Assumption:
Shapiro-Wilk's Test (rankits)
Equality of Variances Assumption:
Levene's Test (rankits)
(ranks)
Null Hypotheses:
Sediment 1 = Reference
LSD Test (rankits)
/-Test (rankits, unequal variances)
Conover T-Test
/-Test (ranks, unequal variances)
Sediment 2 = Reference
LSD Test (rankits)
/-Test (rankits, unequal variances)
Conover T-Test
/-Test (ranks, unequal variances)
Sediment 3 = Reference
LSD Test (rankits)
/-Test (rankits, unequal variances)
Conover J-Test
/-Test (ranks, unequal variances)
Test
Statistic

IT=0.972

/•=0.61
F=1.51


/=3.87
/=4.69
/=4.14
/=6.18

/=3.32
/=3.76
/=3.54
/=3.95

/=1.66
/=1.69
/=1.86
/=1.85
Probability
P

0.791

0.621
0.236


0.0024
0.0011
0.0016
0.0003

0.0053
0.0040
0.0038
0.0046

0.0677
0.0706
0.0497
0.1215

a

0.01

0.10
0.10


0.05
0.05
0.05
0.05

0.05
0.05
0.05
0.05

0.05
0.05
0.05
0.05

Conclusion

do not reject

do not reject
do not reject


reject
reject
reject
reject

reject
reject
reject
reject

do not reject
do not reject
reject
do not reject
Using Eq. 18, the power of the LSD test for detecting a 100% increase in dredged sediment bioaccumulation
over the mean reference bioaccumulation (i.e., d=0.066 (ig/g) can be determined by:

                            /!., = (0.066)/[5/2(0.003763)]* - 1.746 = -0.045

and 1-p for /=-0.045 with  16 df is 0.48.  Power values for 10, 25, 50, 100, 200 and 300% increases over mean
reference bioaccumulation are given in the output for SAS program BIOACC (Section D4.3.2).

The sample size (n) required to provide a 0.95 probability (l-p=0.95) of detecting a 25%  increase (0.0165
jig/g) over the mean reference bioaccumulation, calculated using the z-approximation (Eq.  9) with MSB re-
placing s2, is:
                    n = 2(1.645 + 1.645)2[0.003763/(0.0165)2] -I- 0.25(1.645)2 = 300 !

Using the same equation,  to detect a 100% increase (0.066 jig/g) over the mean reference bioaccumulation
with a power of 0.95, n = 20.  Assuming we are limited to 5 replicates, there is a 0.95 probability of detecting
                                                                                    DRAFT

-------
	D-43	

a difference (d) of 0.135 \ig/g, which is a 205% increase over the mean reference bioaccumulation.  Other
values of d when power = 0.5, 0.6, 0.7,0.8, 0.9, and 0.99 are given in the output for SAS program BIOACC
(Section D4.3.2).
D3.1.2         Comparisons with an Action Level

In this comparison, the objective is to determine whether the mean bioaccumulation of contaminants in
animals exposed to a dredged sediment is significantly less than a specified action level or standard.  If the
mean tissue concentration of one or more contaminants of concern is greater than or equal to the applicable
action level, then no statistical testing is required.  The conclusion would be that the dredged sediment does
not meet the guidelines associated with the action level (Section 6.3).  If the mean tissue concentrations of
a contaminant of concern are less than the applicable action level, then a confidence-interval approach is used
to determine if these means are significantly less than the action level.  One-sided tests are appropriate since
there is concern only if bioaccumulation from the dredged sediment is not significantly less than the  action
level. There are two different approaches to conducting these tests, and both are acceptable.

The first approach is to calculate a value of t, much as in a r-test (this approach is often called a one-sample
f-test):
                                             - action Uvel ^                              (£q  2Q)
where X, s2 and n refer to mean, variance, and number of replicates for contaminant bioaccumulation from
the dredged sediment.

If tests of equality of variances in the comparison of dredged sediments with the reference indicate that vari-
ances are equal for all sediments, then MSE from the ANOVA is used as s\ and calculated t is compared to
tAK, with N - k degrees of freedom.  If the variances are not equal, then s1 for the individual sediment is used,
and calculated t compared with fft9s, with n - 1 degrees of freedom. If the data were log-transformed to nor-
malize the distributions or equalize variances, then all calculations should be carried out on  log-transformed
values.

Another approach is to calculate the upper one-sided 95% confidence limit (UCL), and compare it  to the
action level:
                                                                                    DRAFT

-------
                                               D-44
                                     UCL - x  +
                                                                                   (Eq.21)
As in the first approach, the MSB is used in place of j2 if variances are not significantly different, and the
degrees of freedom (v) are N - k.  If variances are significantly different, s2 for the individual sediment is used,
and v for each sediment i = n, -1.  There is a 0.95 probability that the true population mean tissue level is
below the UCL.  If the UCL is below the action level, there is a 20.95 probability that the population mean
tissue level for the dredged sediment is below the action level, and we conclude that the action level is not
exceeded. If the UCL is above the action level, we cannot be sure that the mean population tissue level does
not exceed the action level.
Either of the above procedures may be used with data that have failed the normality test, but the results
should be considered approximate.

The choice of which approach to use depends on the computer software and the presentation method to be
used.  In SAS, it is more convenient to calculate the UCL and compare with the action level, as in program
BIOACC (Section D4.3).  In SYSTAT, it is simpler  to conduct a one-sample Mest.  Both approaches can
easily be performed by hand.  If the data are presented graphically, as in Figure D-6, the confidence-level
approach is used. If the investigator wants to provide the exact probability that  the mean tissue level is less
than the action level, then the one-sample Mest is used.
          O.24
      ^-  0.22 -
g    0.2-
«§
j»   0.1 a —

J5-  o.ie-
S
CXI
      o>
          O.1 4 -
          0.1 2
                                              UCL-r
                       mean ™
                                                                       Action Level
                                                                          UCL-r
                                                                        mean •
                      Sediment 1
                                               Sediment 2
                                                                        Sediment 3
                                           Dredged  Material
Figure D-6     Comparison of Mean Dredged Sediment Contaminant Tissue Levels (mean) and 95% Upper
               Confidence Level (UCL) with Hypothetical Action Level.

Figure D-6 presents a comparison of mean bioaccumulation from the three dredged sediments (see Table D-9)
with a hypothetical action level of 0.2 jig/g. There is no need to calculate the UCL for sediment 1 as the mean
exceeds the action level.  Because variances were not significantly different for the untransformed data (Table
D-10), we use MS£=0.003763 and t&9iK-l.l46 in Eq. 21 to obtain:
                                                                                   DRAFT

-------
                                               D-45
                             UCL « 0.190 + 1.746(0.003763/5)* = 0.238

for sediment 2, and UCL = 0.178 for sediment 3. SAS program BIOACC (Section D4.3) calculates UCL for
both equal and unequal variances.

If the UCL lies below the action level, there is a >0.95 probability that the true population mean tissue level
for that sediment is  less than the action level. Thus, we would conclude that mean bioaccumulation for
dredged sediment 3 is less than the action level.  Because the UCL for sediment 2 exceeds the action level even
though the sample mean does not,  we cannot be sure that the true population mean tissue level for this
sediment is less than the action level.

Formulae for calculating statistical power for comparisons to a fixed standard such as an action level are very
similar to Eqs. 8 and 9:
                                                                                          (Eq>22)
where sz and v (degrees of freedom) are MSE and N - k if variances are equal (or expected to be equal, if the
calculation is made prior to testing), and s2 for the individual sediment and n-, - 1 if variances are unequal.
It is usually easier to use the z-approximation (from Alldredge, 1987) to avoid solving for n iteratively:
                                                                                              23)
The formulae indicate that the sample size required to detect any given difference d will be approximately one-
half that required for a comparison of two treatments. The sample size required is lower because the com-
parison is made to a fixed value, rather than to a reference which can also vary. Thus, there is no sampling
uncertainty or error for  the fixed standard and we know the true value of one of the two  things being
compared.

Using the z-approximation and s2=MSE, the sample size required to provide a 0.95 probability (1-P=0.95) of
detecting a tissue level 25% (0.05 \ig/g) below the action level is:

                      n = (1.645 + 1.645)2(0.003763)/0.0025 + 0.5(1.645)2 = 18.

The minimum significant difference is:
                                                                                    DRAFT

-------
                                               D-46

                                         * = 1.746(0.003763/5)* = 0.048 |ig/g.
The power of a comparison can be determined by:
When  variances are not significantly different,  s  is replaced by (MSE)*  and v  = N - k  df.  Using
M5£=0.003763 as above, the power to detect a 10% decrease in mean bioaccumulation below the action level
is 0.16, and power to detect a 50% decrease is 0.96.  Power for 10,20,30,40 and 50% decreases are given in
the output for SAS program BIOACC (Section D4.3.2).
D3.2           Tier IV Time-Sequenced Laboratory Bioaccumulation Study

The time-sequenced laboratory bioaccumulation test in Tier IV is designed to detect differences, if any,
between steady-state bioaccumulation in organisms exposed to the dredged sediments and steady-state bioac-
cumulation in organisms exposed to the reference sediment. If organisms are exposed to biologically available
contaminants under constant conditions for a sufficient period of time, bioaccumulation will eventually reach
a steady state in which maximum bioaccumulation has occurred, and the net exchange of contaminant between
sediment and organism is zero.

A simple kinetic model (McFarland and Clarke, 1987; Clarke and McFarland, 1991) can be used with data col-
lected over a relatively short period of constant exposure to project tissue concentrations at steady state. This
model integrated for constant exposure  is:
                                                                                         (Eq-25)
where  C, = concentration of a compound in tissues of an organism at time t,
        kr = uptake rate constant,
        Cw - exposure (water) concentration of the compound,
        k2 =t elimination rate constant, and
        t - time in days.

Using this model, contaminant uptake occurs rapidly at first, and then the rate of uptake gradually diminishes
as uptake begins to level off and approach an asymptote (steady state).

As duration of exposure increases, the exponential term in the model approaches zero, and the tissue con-
                                                                                    DRAFT

-------
                                                D-47

centration at steady state (i.e., infinite exposure) is calculated as:
                                         C, =   -l  . C.  ,                                 (Eq- 26)
where €„ is an estimate of the whole-body concentration of the compound at steady state.

Steady-state concentration estimates from organisms exposed to dredged sediments are compared to applicable
action levels and to steady-state estimates from organisms exposed to the reference sediment. The data analy-
sis involves several steps:

1.      Calculate a separate nonlinear regression for each replicate using Eq. 25.

2.      Use the regression coefficients (ki and fc2) to calculate the steady-state concentrations (CM)
        from Eq. 26, or set up the regression analysis to estimate/output CM directly (see below).

3.      Use the estimates of Cu as data in a statistical test comparing each dredged sediment to  the
        reference (as in Section D3.1.1). Conclusions possible from these comparisons and evaluative
        factors that should be assessed are detailed in Section 6.3.

4.      Use confidence intervals  or one-sample Mests  to compare the  steady-state estimates with
        applicable action levels (as in Section D3.1.2).
D3.2.1     Calculating Steady-State Concentrations

Table D-12 presents example data resulting from a hypothetical 28-day time-sequenced laboratory bioac-
cumulation test using three dredged sediments and a reference sediment.  There are five replicates of each
treatment, and tissue samples were analyzed on days 2, 4, 7, 10, 18, and 28 of the test.  More sampling days
are scheduled in  the early part of the test  to enable more accurate characterization of the early, rapidly
changing portion  of the uptake curve.

These data can be used with iterative nonlinear regression methods such as those in the SAS NLIN or
SYSTAT NONLIN procedures to solve for the parameters (A^ and k2) in the model above.  Then CM, the
steady-state concentration,  is  simply  kfjkj.    In this  iterative  calculation method, the  contaminant
concentration in the sediment (C,), or even a constant such as 1, may be used instead of Cv This is because
the values of the  rate constants and the exposure concentration are not of interest in this application, only
their ratio (i.e., CM). Thus, the equation could be written as:
                                                                                     DRAFT

-------
	D-48	

Table D-12.    Results from a Hypothetical Time-Sequenced Bioaccumulation Test, Showing Contaminant
               Concentrations (ng/g) in Tissues of Animals Exposed to Different Treatments.

Replicate
1





2





3





4





5






Day
2
4
7
10
18
28
2
4
7
10
18
28
2
4
7
10
18
28
2
4
7
10
18
28
2
4
7
10
18
28
Mean Sediment
Concentration
Treatment
Reference
0.054
0.441
0.687
0.037
0.856
0.514
0.163
0.797
0.177
0.549
0.598
0.839
0.391
0.203
0.862
0.884
0.016
0.793
0.234
0.564
0.413
0.787
0.806
0.899
0.034
0.018
0.029
0.294
0.119
0.226

0.45
Sediment 1
0.159
0.516
0.881
0.278
0.904
0.172
0.292
0.158
0.317
0.485
1.300
1.049
0.428
0.743
0.270
0.051
0.671
0.476
0.558
0.324
0.562
0.909
0.934
0.712
0.256
0.126
0.603
0.718
1.173
1.245

4.0
Sediment 2
0.869
0.838
1.246
1.767
1.631
1.178
0.726
0.633
0.816
1.272
1.877
1.721
0.394
0.452
0.897
1.003
1.487
1.366
1.232
0.728
1.639
1.158
1.216
1.513
0.977
1.314
0.688
1.415
1.280
1.843

33.0
Sediment 3
0.745
1.316
1.583
1.578
2.822
1.295
1.703
0.930
2.715
2.268
2.607
2.964
2.045
2.141
1.016
1.756
3.414
2.109
1.855
1.150
2.221
2.899
1.319
2.820
1.135
1.621
2.134
0.890
1.866
3.325

44.0
and CM estimated directly by the regression software.  The estimate of Ctt should be the same regardless of
which approach is used. SAS program BIOACCSS (Section D4.4) performs the steady-state calculations using
C., and outputs the regression parameters and Cu for each replicate to a new data set.  These are displayed
in Table D-13.
                                                                                   DRAFT

-------
                                               D-49
Table D-13.    Regression Parameters Estimated from Example Time-Sequenced Bioaccumulation Data.
Treatment
Reference






Sediment 1






Sediment 2






Sediment 3






c. ug/g
0.45






4.0






33.0






44.0






Replicate
1
2
3
4
5


1
2
3
4
5


1
2
3
4
5


1
2
3
4
5


*i
0.237
0.306
0.540
0.318
0.045


0.059
0.019
0.243
0.051
0.024


0.014
0.007
0.006
0.034
0.023


0.011
0.015
0.094
0.024
0.008


*2
0.176
0.201
0.407
0.162
0.087


0.427
0.047
2.206
0.243
0.060


0.319
0.113
0.120
0.878
0.568


0.250
0.236
1.977
0.458
0.139


cu
0.608
0.687
0.597
0.883
0.234
Mean = 0.602
SE = 0.105
0.554
1.644
0.441
0.833
1.600
Mean = 1.014
SE = 0.256
1.488
1.907
1.511
1.290
1.350
Mean = 1.509
SE = 0.108
1.964
2.776
2.087
2.259
2.648
Mean = 2.347
SE = 0.158
Nonlinear regressions for the example data were calculated using the SAS NLIN procedure with the DUD
method.  This method does not require derivatives. Other methods may be used but the derivatives for the
parameters (fcj and k2, or-CM and k2 if Ca is estimated directly) must be specified.  The Marquardt method and
the Gauss-Newton method produced results similar to DUD for the example data.

Iterative  curve-fitting techniques will provide better fits to some data than to others, and the asymptotic
relationship will not always be the best fit to the data.  Thus, investigators should be aware of the following
problems:

1.      Failure to converge on a solution within the allowed number of iterations. Always have the regression
       software print out the results, even though the regressions are only used to create a new data set of
       CM values. SAS will output the parameter estimates from the final iteration, regardless of whether
       convergence occurred. If the last few iterations approach convergence (i.e., there is little change in
                                                                                    DRAFT

-------
	D-SO	

        parameter estimates and residual error mean square), then the parameter estimates from the last
        iteration may be used. If convergence was not approached, then the program should be run again for
        that replicate, using the parameter values from the last iteration as starting values.

2.      No relationship between concentration and time.  This can occur in sediments with low or non-
        detectable contaminant levels. The model-derived estimate of Cu will usually converge on the mean
        tissue concentration over all days.

3.      A non-asymptotic relationship. If the relationship between tissue levels and time is linear, rather than
        asymptotic, the estimated asymptote  (CH) will approach infinity.  A linear  relationship will occur if
        the experiment was not  conducted for a long enough time for  the  tissue levels to approach the
        asymptote, or because of anomalously high tissue levels later in the experiment.  Always plot the data
        prior to calculating the regressions to make sure the relationships are asymptotic.   SAS program
        BIOACCSS (Section D4.4) provides separate plots for each treatment, with the replicates identified.
        Anomalies/outliers and non-asymptotic relationships for any replicate can easily be spotted using plots
        such as these.

        If relationships for only  one or a few replicates are non-asymptotic, then those  replicates can be
        dropped from the analysis, or the maximum measured tissue concentrations used  as an estimate of
        Cu. If relationships for several replicates (i.e., >5 total, or >1 for any individual sediment) are non-
        asymptotic, then there is little justification for assuming that a steady state has been approached.  The
        test should be repeated, but over a longer time interval. Measuring concentrations in field-collected
        organisms is also an alternative, if steady state is not reached in laboratory  experiments (see Section
        D3.3).

4.      Estimates of Cu that are negative.  This can happen if tissue concentrations decrease over time and
        k2 is negative. If there are only one or a few replicates with negative CM values,  then these replicates
        can be dropped from the analysis. Alternatively, the minimum or mean measured concentration could
        be used as an estimate of Ca. If there are several (i.e., >5 total, or >1 for  any individual sediment),
        then the test should be repeated.  High initial contaminant levels in the test organisms are the most
        probable cause of negative values of CM values. Prior to repeating the test,  these initial contaminant
        levels should be measured, and the source of test  organisms should  be changed if these levels are
        greater than bioaccumulation of the contaminant at the end  of the previous test.

If difficulties are encountered, approaches such as those discussed by Draper and Smith (1981) and SCI (1989)
should be considered.  Investigators with  limited experience should always  consult  an applied  statistician
familiar with nonlinear regression prior to analyzing time-sequenced bioaccumulation data.  It is important
to remember that these data are usually very expensive to obtain, because of the extensive number of chemical
analyses required, and the data should be carefully and correctly analyzed.

In the example data analysis, the DUD method failed to converge within the default  number of iterations (50)
                                                                                      DRAFT

-------
	D-51	

for sediment 3, replicate 5. However, the procedure was close enough to convergence that the regression
coefficients output at the final iteration produced a reasonable estimate of Ca.

The approach recommended in this Appendix for comparison of Tier IV dredged sediment and reference
sediment bioaccumulation data differs from that described in the Ocean Disposal Manual (the "Green Book").
The approach of comparing 95% confidence intervals for Cu is not recommended because:

        •  The 95% confidence intervals apply to the estimate of CM rather than to the difference
           between estimates

        •  The 95% confidence intervals are based on regressions through points from all repli-
           cates for a treatment, ignoring variation among replicates within a treatment

        •  Different  programs or methods will provide different confidence intervals for the
           same data

        •  Measurements of tissue levels taken at different times may not be independent.

If the objective of the Tier IV investigation is only to compare bioaccumulation from reference and dredged
sediments over the duration of the experiment, and estimates  of Cu are not required, there are other alterna-
tives to analyze the data:

        •  Repeated measures analysis of variance, testing for linear and quadratic components
           of the time trend

        •  Multivariate analysis of variance (MANOVA), with tissue levels for each day consid-
           ered separate variables (linear and quadratic trends can also be tested in MANOVA).

These alternatives are equivalent with respect to testing for linear and quadratic trends over time, and some
repeated measures programs (e.g., SYSTAT MGLH) will provide MANOVA results as well.  These alterna-
tives should only be used by experienced investigators who are familiar with them.  Both alternatives would
be most useful in testing for an overall quadratic trend, as the absence of such a trend over time would indi-
cate that tissue levels  did not approach an asymptote within the duration of the experiment.
D3.2.2     Comparison with Reference Sediments and Action Levels

The difficult part of analyzing time-sequenced bioaccumulation tests is obtaining sound estimates of Cu. Once
these estimates are obtained, they are analyzed using the same procedures as for single time-point bioaccumu-
lation tests (Section D3.1).  Steady-state concentration estimates for the dredged sediments are compared to
steady-state concentration estimates for the reference sediment using the appropriate methods from the deci-
                                                                                   DRAFT

-------
	D-52	

sion trees in Figures D-5A or 5B.

The values of Cu in Table D-13 were analyzed using the decision tree steps in Figure D-SA. Although SAS
Program BIOACCSS (Section D4.4) conducts all parametric and rankit analyses from the decision trees, only
the appropriate results are reported in Table D-14. The untransformed Cu values were normally distributed
(Shapiro-Wilk's Test, P>0.01, the a level from Table D-2 for N=2JO, balanced data).  However, neither the
untransformed nor log-transformed CM passed Levene's Test for equality of variances (J°<0.10, the a level in
Table D-2 for n=5, balanced data). Therefore, Mests were conducted, comparing each dredged sediment CH
with reference sediment Ctt, using the untransformed Cu estimates. Note that Mests for equal variances could
be used because the F' tests for each dredged sediment-reference comparison did not reject equal  variances,
even though the overall test of equality of variances indicated unequal variances within the data set as a whole.
Mean estimated concentrations at steady state for dredged  sediments  2 and 3 (but not sediment  1) were
significantly greater than that of the reference sediment (Table D-14).

Power calculations for an LSD test using untransformed  data are performed in SAS program BIOACCSS
(Section D4.4). From Eq. 18, a 50% increase over the  mean reference CM (0.602 ng/g) can be detected with
a probability of 0.32, and a 100% increase with a probability of 0.78.  Likewise, there is a 0.95 probability of
detecting a 138% increase in CM over the mean reference CM. The least significant difference from the LSD
is 0.415 |ig/g, which is a 69% increase over the mean reference Ca. Sample size (n) required to provide a 0.95
probability of detecting a 25% increase over the mean reference Cu is 136 (Eq. 9, using MSE in place of s2).

The CM values for the dredged sediment  can also be compared to an action level, if available, using the one-
sample Mest or one-sided upper confidence limits (UCL) as in Section D3.1.2.  UCL for both equal variances
and unequal variances may be calculated using SAS program BIOACCSS (Section D4.4). Figure D-7 provides
the mean CM and UCL for each example dredged sediment, along with a hypothetical  action level of 2 jig/g.
The UCL for sediments 1 and 2 were below the action  level, indicating that the CM for these sediments were
significantly lower than the action level. The mean Ca for sediment 3 was above the action level, so there was
no need to calculate a  UCL to conclude that the Cn was not significantly lower than the action level.

Power to detect a true population steady-state concentration 10, 20,  30, 40 and 50% below an action level is
calculated in SAS program BIOACCSS (Section D4.4).
D3.3       Steady-State Bioaccumulation from Field Data

The field bioaccumulation test is designed to show differences, if any, between organisms living at the proposed
disposal site and organisms living in the sediments in the reference area.  This approach is valid only under
the conditions described in Section 12.2.2.

Replicate tissue concentrations in organisms collected at the disposal site(s) are compared with replicate tissue
                                                                                    DRAFT

-------
                                         D-53
Table D-14.   Tests of Assumptions and Parametric Hypothesis Tests on Untransformed Steady-State Bio-
             accumulation Example Data.
Null Hypothesis: Mean Dredged Sediment Steady-State Bioaccumulation Equals Mean Reference
Sediment Steady-State Bioaccumulation
Test
Normality Assumption:
Shapiro- Wilk's Test
Untransformed data
Log-transformed data
Equality of Variances Assumption:
Levene's Test
Untransformed data
Log-transformed data
Null Hypotheses:
Sediment 1 = Reference
f-Test (equal variances)
Sediment 2 = Reference
/-Test (equal variances)
Sediment 3 = Reference
f-Test (equal variances)
Test
Statistic
W=Q.9&
W=0.943
F=4.74
F=3.68
f=1.49
f=6.03
f=9.21
Probability
P
0.613
0.280
0.015
0.034
0.0873
0.0002
<0.0001
a
0.01
0.01
0.10
0.10
0.05
0.05
0.05
Conclusion
do not reject
do not reject
reject
reject
do not reject
reject
reject
        2.4-
        2.2-
   •
co-   1-8H
CO
O
I   1-«H
2
     1.4-
        1.2-
                                                            mean
                                                             Action Level
                 UCL -r
               mean
                                        UCL-r
                                      mean i
                 Sediment 1
                                    Sediment  2.
                                    Dredged Material
Sediment 3
Figure D-7.
          Comparison of Mean Dredged Sediment Contaminant Steady-State Tissue Levels (CH) (mean)
          and 95% Upper Confidence Levels (UCL) with Hypothetical Action Level.
                                                                         DRAFT

-------
	D-54	

concentrations in organisms collected from the reference area using the decision tree steps in Figures D-SA
and SB. If comparisons involve organisms from only one disposal site, then the appropriate statistical com-
parison procedures, depending on the results of the tests of assumptions, are the two-sample Mest for equal
or unequal variances, or the Mest for unequal variances using rankits or ranks (Section D2.1.1.1).
D4.0       SAS PROGRAMS AND OUTPUT FOR EXAMPLE DATA

This Section provides SAS programs to analyze the example data sets given in Appendix D.  Each program
includes all analyses from the corresponding decision tree that would be performed using SAS.  While it is
certainly possible to conduct the statistical analysis of a data set in a stepwise fashion, we find it much more
efficient to perform all analyses at once, and then select the appropriate results based on the steps in the
decision tree.  Power calculations are provided in addition to the decision tree analyses.

SAS statements in the sections that follow are given in uppercase letters (although this is not required for
SAS).  Comments within the body of the programs are in upper and lowercase letters in the following format:
/* Comment line. */. Every SAS statement must end with a semicolon, but several statements may be included
on the same line. The programs are designed for the analysis of Appendix D example data, but can be used
with other data sets after minor modifications.  Investigators wishing to use these programs should have some
familiarity with SAS. SAS output follows each program; the output has been edited to remove much of the
nonessential information.

We recommend that data analysis reports include at least the following:

        •   Number of replicates, mean and SE for each  treatment

        •   Treatment of less-than detection limit data, if any

        •   Results of tests of assumptions

        •   Data transformation used, if any

        •   Name of statistical hypothesis testing procedure, its calculated test statistic and associated
           probability, and conclusion reached regarding the null hypothesis

        •   Minimum significant difference or some other indication of power for a parametric LSD
           test or Mest.
                                                                                    DRAFT

-------
                                           D-55

D4.1         Program WATTOX.SAS for Water Column Toxicity Test Data Analysis


WATTOX.SAS is a program to compare dilution water survival vs. 100% elutriate survival, using an arcsine-square
root transformation on the data. The program performs all statistical analyses in Figure D-l. Included in these
analyses are: mean survival for dilution water and elutriates, Shapiro-Wilk's Test for normality, Mest for equal or
unequal variances, and a Mest for unequal variances on data converted to rankits.  Refer to the decision tree in
Figure D-l to determine which test results should be used.  Minimum significant difference and some other power
calculations for the parametric /-test are also provided.
D4.1.1        WATTOX.SAS Program Statements


LIBNAME  Q 'C:\SAS';
OPTIONS  LINESIZE=79  PAGESIZE=59 NODATE NONUMBER;

/* Identify the treatment codes.  */

PROC FORMAT;
 VALUE TRTFMT
   0='DILUTION WATER
   1='100% ELUTRIATE
   2='50% ELUTRIATE
   3='25% ELUTRIATE
   4='12.5% ELUTRIATE

/* Input the toxicity test data after the CARDS  statement,  listing the       */
/* treatment code, replicate,  and number of  survivors.  A permanent SAS      */
/* data  set is created in the  directory specified in the LIBNAME statement.  */

DATA Q.WATCOL;
 INPUT TRT REP SURV  @@;
 CARDS"
0 1 2o'o 2 19 0 3  20 0 4 20 0  5 19
116127139145158
21822823924 10 25 11
3 1 12 3 2 18 3 3  15 3 4 14 3  5 13
4 1 17 4 2 17 4 3  18 4 4 16 4  5 18
i
/* Input no. of organisms (M)  per test container  at start of test. */
/* Calculate proportion of survivors (SURV/M)  and take the  SQRT. */
/* Arcsine transform SQRT(SURV/M). */
/* Format, print,  sort the data.   Print no.  of observations,  mean, and */
/* standard error  for survival in each treatment. */

DATA AO;
 SET Q.WATCOL;
 M=20;
 ARCSURV=ARSIN(SQRT(SURV/M));
 LABEL TRT='TREATMENT GROUP'
       REP='REPLICATE'
       M='NO. OF ORGANISMS PER REPLICATE'
       SURV='NUMBER  OF SURVIVORS'
       ARCSURV='ARCSINE TRANSFORMATION';
 FORMAT  TRT TRTFMT.;
 TITLE  'WATER COLUMN TOXICITY  DATA';
PROC PRINT LABEL;  VAR TRT REP  M SURV ARCSURV;
                                                                        DRAFT

-------
                                       D-56

PROC SORT; BY TRT;
PROC MEANS NOPRINT; BY TRT; VAR SURV;
 OUTPUT OUT=Y N=N SUM=TOTAL MEAN=MEANSURV STDERR=SE;
PROC PRINT LABEL; VAR TRT N MEANSURV SE;
 LABEL MEANSURV='MEAN SURVIVAL';

/* Delete data not needed for the dilution water-100% elutriate comparison. */
/* Print descriptive statistics. */

DATA A;
 SET AO;
 IF TRT>1 THEN DELETE;
 TITLE2 'ARCSINE-SQUARE ROOT TRANSFORMATION';
PROC MEANS NOPRINT; VAR ARCSURV; BY TRT; ID M;
 OUTPUT OUT=X N=N MEAN=MEAN VAR=VARIANCE STD=S STDERR=SE;
PROC PRINT LABEL; VAR TRT N MEAN VARIANCE S SE;

/* Test normality of residuals using Shapiro-Wilk's Test. */

PROC GLM DATA=A NOPRINT;
 CLASS TRT;
 MODEL ARCSURV=TRT;
 OUTPUT OUT=Z R=RESID;
PROC UNIVARIATE NORMAL DATA=Z;
 VAR RESID;
 TITLE3 'SHAPIRO-WILKS TEST';

/* Conduct t-test, which includes F' test for equality of variances. */

PROC TTEST DATA=A;
 CLASS TRT;
 VAR ARCSURV;
 TITLE3 'T-TEST';

/* Convert data to rankits and conduct t-test. */

PROC RANK DATA=A NORMAL=BLOM OUT=A1;
 VAR SURV; RANKS RANKIT;
PROC TTEST DATA=A1;
 CLASS TRT;
 VAR RANKIT;
 TITLE2 'DATA CONVERTED TO RANKITS';

/* Calculate minimum significant difference and power of a t-test to detect */
/* true population differences of 10, 20, 30, 40 and 50% below mean */
/* dilution water survival. */

DATA BO;
 MERGE X Y;
 IF TRTA=0 THEN DELETE;
 MEANO=MEAN; NO=N; S20=VARIANCE;
 MEANPCT=MEANSURV/M;
DATA Bl;
 SET X;
 IF TRTA=1 THEN DELETE;
 N1=N; S21=VARIANCE;
DATA B2;
 MERGE BO Bl;
 DF=NO+Nl-2;
 N=(NO+Nl)/2;
 S2POOL=(S20*(NO-1)+S21*(N1-1))/DF;
 TALPHA=TINV(.95,DF);
 DMIN=TALPHA*SQRT(2*S2POOL/N);
                                                                   DRAFT

-------
                                       D-57
 LABEL N='NO. OF REPLICATES'
       MEANPCT='MEAN DILUTION WATER SURVIVAL'
       S2POOL='POOLED VARIANCE'
       DF='DEGREES OF FREEDOM, DF'
       TALPHA='T VALUE FOR  (1-ALPHA=0.95,DF)'
       DMIN='MINIMUM SIGNIFICANT DIFFERENCE';
 TITLE2 'POWER OF T-TEST TO DETECT A TRUE POPULATION DIFFERENCE (D)';
 TITLE3 'FROM MEAN DILUTION WATER SURVIVAL USING ARCSINE TRANSFORMATION';
PROC PRINT LABEL NOOBS; VAR M N MEANPCT S2POOL DF TALPHA DMIN;
DATA B3;
 SET B2;
 DO PCTDIFF=10 TO 50 BY 10;
  SEDSURV=MEANPCT-PCTDIFF/100;
  ARCSURV=ARSIN(SQRT(SEDSURV));
  ARCDIFF=MEANO-ARCSURV;
  TBETA=(SQRT(N)*ARCDIFF)/SQRT(2*S2POOL)-TALPHA;
  POWER=PROBT(TBETA,DF);
  OUTPUT;
  END;
 LABEL PCTDIFF='% REDUCTION IN SURVIVAL FROM DIL. WATER'
       SEDSURV='100% ELUTRIATE SURVIVAL'
       ARCSURV='ARCSINE 100% ELUTRIATE SURVIVAL'
       ARCDIFF='D'
       TBETA='T VALUE FOR  (1-BETA,DF)';
PROC PRINT LABEL;
 VAR PCTDIFF SEDSURV ARCSURV ARCDIFF TBETA POWER;
 TITLE;
D4.1.2
WATTOX.SAS Program Output
                            WATER COLUMN TOXICITY DATA


OBS
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24


TREATMENT GROUP
DILUTION WATER
DILUTION WATER
DILUTION WATER
DILUTION WATER
DILUTION WATER
100% ELUTRIATE
100% ELUTRIATE
100% ELUTRIATE
100% ELUTRIATE
100% ELUTRIATE
50% ELUTRIATE
50% ELUTRIATE
50% ELUTRIATE
50% ELUTRIATE
50% ELUTRIATE
25% ELUTRIATE
25% ELUTRIATE
25% ELUTRIATE
25% ELUTRIATE
25% ELUTRIATE
12.5% ELUTRIATE
12.5% ELUTRIATE
12.5% ELUTRIATE
12.5% ELUTRIATE


REPLICATE
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
NO. OF
ORGANISMS
PER
REPLICATE
20
20
20
20
20
20
20
20
20
20
20
20
20
20
20
20
20
20
20
20
20
20
20
20
NUMBER
OF
SURVIVORS
20
19
20
20
19
6
7
9
5
8
8
8
9
10
11
12
18
15
14
13
17
17
18
16

ARCSINE
TRANSFORMATION
1.57080
1.34528
1.57080
1.57080
1.34528
0.57964
0.63305
0.73531
0.52360
0.68472
0.68472
0.68472
0.73531
0.78540
0.83548
0.88608
1.24905
1.04720
0.99116
0.93774
1.17310
1.17310
1.24905
1.10715
                                                                   DRAFT

-------
                                       D-58
 25
12.5% ELUTRIATE
              DBS    TREATMENT GROUP

               1     DILUTION WATER
               2     100% ELUTRIATE
               3     50% ELUTRIATE
               4     25% ELUTRIATE
               5     12.5% ELUTRIATE
                                 N
   20
       MEAN
     SURVIVAL
   18
1.24905
       SE
5
5
5
5
5
19.6
7.0
9.2
14.4
17.2
0.24495
0.70711
0.58310
1.02956
0.37417
                            WATER COLUMN TOXICITY DATA
                        ARCSINE-SQUARE ROOT TRANSFORMATION
            TREATMENT
   OBS        GROUP         N      MEAN     VARIANCE

    1     DILUTION WATER    5    1.48059    0.015257
    2     100% ELUTRIATE    5    0.63126    0.006986
                                                 0.12352
                                                 0.08358
                              SE

                           0.055239
                           0.037379
Variable=RESID
Variable: ARCSURV

      TRT
                            WATER COLUMN TOXICITY DATA
                        ARCSINE-SQUARE ROOT TRANSFORMATION
                                SHAPIRO-WILKS TEST
                               UNIVARIATE PROCEDURE
                   N
                   W:Normal
                             10
                       0.846238
Prob j T |
0.0001
0.0000
0.12351878
0.08358232

0.05523928
0.03737915

For HO: Variances are equal, F' =2.18    DF =  (4,4)
                                                 Prob>F'
                          0.4679
                                                                   DRAFT

-------
                                            D-59
Variable: RANKIT

       TRT
                  WATER COLUMN  TOXICITY  DATA
                   DATA CONVERTED TO RANKITS

                        TTEST PROCEDURE

             RANK FOR  VARIABLE SURV

          N              Mean                Std Dev
                                                     Std Error
DILUTION WATER
100% ELUTRIATE
Variances
Unequal 4 .
Equal 4.
T
6306
6306
5
5

DF
7.7
8.0
0.74011839 0.44830825
-0.74011839 0.55672332
Prob> T|
0.0019
0.0017
0.
0.

20048954
24897424

For  HO:  Variances are equal,  F'  = 1.54
                                   DF =  (4,4)
                                           Prob>F'  = 0.6850
                                WATER COLUMN TOXICITY DATA
              POWER OF T-TEST  TO DETECT A TRUE  POPULATION DIFFERENCE  (D)
           FROM MEAN  DILUTION WATER  SURVIVAL USING ARCSINE TRANSFORMATION
   NO.  OF
 ORGANISMS
     PER
 REPLICATE

      20
    N

    5
  MEAN               DEGREES
 DILUTION               OF
WATER      POOLED   FREEDOM,
 SURVIVAL  VARIANCE     DF
                            MINIMUM
          T VALUE FOR     SIGNIFICANT
       (1-ALPHA=0.95,DF)   DIFFERENCE
   0.98   0.011121
8
1.85955
0.12403
OBS

 1
 2
 3
 4
 5
% REDUCTION
IN SURVIVAL
FROM OIL.
WATER
10
20
30
40
50

100%
ELUTRIATE
SURVIVAL
0.88
0.78
0.68
0.58
0.48
ARCSINE
100%
ELUTRIATE
SURVIVAL
1.21705
1.08259
0.96953
0.86574
0.76539



D
0.26354
0.39800
0.51106
0.61485
0.71520

T VALUE
FOR
(1-BETA,DF)
2.09166
4.10768
5.80277
7.35888
8.86344



POWER
0.96508
0.99830
0.99980
0.99996
0.99999
D4.2
Program BENTOX.SAS for Benthic Toxicity Test Data Analysis
BENTOX.SAS is a program to compare benthic toxicity data from dredged sediments vs. reference sediment,
using an arcsine-square root transformation on the data. Included in these analyses are: mean survival from
each sediment exposure, Shapiro-Wilk's Test for normality, Levene's test for equality of variances, r-tests for
equal or unequal variances,  LSD test,  and tests on rankits (normalized ranks for survival).  Refer to the
decision tree in Figures D-4A and 4B to determine which test results should be used.  The program includes
power calculations (on an arcsine-transformed scale) for an LSD test.
                                                                          DRAFT

-------
                                        D-60

D4.2.1       BENTOX.SAS Program Statements


LIBNAME Q 'C:\SAS';
OPTIONS LINESIZE=79 PAGESIZE=59 NODATE NONUMBER;

/* Identify the treatment codes. */

PROC FORMAT;
 VALUE TRTFMT
   1='REFERENCE '
   2='SEDIMENT 1'
   3='SEDIMENT 2'
   4='SEDIMENT 3';

/* Input the toxicity test data after the CARDS statement, listing the  */
/* treatment code, replicate, and number of survivors.  A permanent SAS */
/* data set is created in the directory specified in the LIBNAME statement.  */

DATA Q.BENTHIC;
 INPUT TRT REP SURV @@;
 CARDS *
1 1 2o'l 2 20 1 3 19 1 4 19 1 5 20
2 1 17 2 2 16 2 3 18 2 4 17 2 5 15
3 1 15 3 2 16 3 3 13 3 4 17 3 5 11
4 1 17 4 2 12 4 3 10 4 4 16 4 5 13

•
,
/* Input no. of organisms (M) per test container at start of test. */
/* Calculate proportion of survivors  (SURV/M) and take the SQRT.*/
/* Arcsine transform SQRT(SURV/M). */
/* Format, print, sort the data.  Print no. of observations, mean, and */
/* standard error for survival in each treatment. */

DATA AO;
 SET Q.BENTHIC;
 M=20;
 ARCSURV=ARSIN(SQRT(SURV/M));
 LABEL TRT='TREATMENT GROUP'
       REP='REPLICATE'
       M='NO. OF ORGANISMS PER REPLICATE'
       SURV='NUMBER OF SURVIVORS'
       ARCSURV='ARCSINE TRANSFORMATION';
 FORMAT TRT TRTFMT.;
 TITLE 'BENTHIC TOXICITY DATA';
PROC RANK NORMAL=BLOM OUT=A;
 VAR SURV; RANKS RANKIT;
PROC PRINT LABEL; VAR TRT REP M SURV ARCSURV RANKIT;
 LABEL RANKIT='NORMALIZED RANK FOR SURVIVAL';
PROC SORT; BY TRT;
PROC MEANS NOPRINT; BY TRT; VAR SURV; ID M;
 OUTPUT OUT=Y N=N SUM=TOTAL MEAN=MEANSURV STDERR=SE;
PROC PRINT LABEL; VAR TRT N TOTAL MEANSURV SE;
 LABEL MEANSURV='MEAN SURVIVAL';

/* Print descriptive statistics for the arcsine-transformed survival data.  */

PROC MEANS NOPRINT DATA=A; VAR ARCSURV; BY TRT;
 OUTPUT OUT=X N=N MEAN=MEAN VAR=VARIANCE STD=S STDERR=SE;
 TITLE2 'ARCSINE-SQUARE ROOT TRANSFORMATION';
PROC PRINT LABEL; VAR TRT N MEAN VARIANCE S SE;

/* Test normality of residuals using  Shapiro-Wilk's Test. */
                                                                   DRAFT

-------
                                        D-61

PROC GLM DATA=A NOPRINT;
 CLASS TRT;
 MODEL ARCSURV=TRT;
 OUTPUT OUT=Z R=RESID;
PROC UNIVARIATE NORMAL DATA=Z;
 VAR RESID;
 TITLE3 'SHAPIRO-WILKS TEST FOR NORMALITY';

/* Conduct Levene's Test for equality of variances. */

DATA AX;
 MERGE AX; BY TRT;
 ABSDEV=ABS(ARCSURV-MEAN);
 LABEL ABSDEV='ABSOLUTE DEVIATIONS FROM MEAN';
PROC GLM;
 CLASS TRT;
 MODEL ABSDEV=TRT;
 TITLE3 'LEVENE"S TEST FOR EQUALITY OF VARIANCES';

/* Perform LSD Test. */

PROC GLM DATA=A OUTSTAT=W;
 CLASS TRT;
 MODEL ARCSURV=TRT;
 MEANS TRT/LSD ALPHA=.l;
 TITLES 'LSD TEST';

/* Perform t-tests for each dredged sediment-reference sediment comparison. */

DATA Tl;
 SET A;
 IF TRT>2 THEN DELETE;
PROC TTEST;
 CLASS TRT;
 VAR ARCSURV;
 TITLE3 'T-TEST';
DATA T2;
 SET A;
 IF TRT=2 OR TRT=4 THEN DELETE;
PROC TTEST;
 CLASS TRT;
 VAR ARCSURV;
DATA T3;
 SET A;
 IF TRT=2 OR TRT=3 THEN DELETE;
PROC TTEST;
 CLASS TRT;
 VAR ARCSURV;

/* Test normality and equality of variances of rankits. */

PROC GLM NOPRINT DATA=A;
 CLASS TRT;
 MODEL RANKIT=TRT;
 OUTPUT OUT=Z1 R=RESID;
 TITLE2 'SURVIVAL DATA CONVERTED TO RANKITS';
PROC UNIVARIATE NORMAL;
 VAR RESID;
 TITLE3 'SHAPIRO-WILKS TEST FOR NORMALITY';
PROC MEANS DATA=A NOPRINT;
 BY TRT; VAR RANKIT;
 OUTPUT OUT=X1 MEAN=MEAN;
DATA AX1;
                                                                   DRAFT

-------
                                        D-62

 MERGE A XI; BY TRT;
 ABSDEV=ABS(RANKIT-MEAN);
 LABEL ABSDEV='ABSOLUTE DEVIATIONS FROM MEAN';
PROC GLM;
 CLASS TRT;
 MODEL ABSDEV=TRT;
 TITLES 'LEVENE"S TEST';

/* Perform LSD test on rankits. */

PROC GLM DATA=A;
 CLASS TRT;
 MODEL RANKIT=TRT;
 MEANS TRT/LSD ALPHA=.l;
 TITLE3 'LSD TEST ON RANKITS';

/* Perform t-tests comparing each dredged sediment with the reference */
/* using rankits. */

PROC TTEST DATA=T1;
 CLASS TRT;
 VAR RANKIT;
 TITLES 'T-TEST ON RANKITS';
PROC TTEST DATA=T2;
 CLASS TRT;
 VAR RANKIT;
PROC TTEST DATA=T3;
 CLASS TRT;
 VAR RANKIT;


/* Calculate power of an LSD test to detect true population differences */
/* of 10, 20, 30, 40 and 50% below mean (arcsine-transformed) reference */
/* sediment survival. */

DATA Cl;
 SET W;
 IF _TYPE_A='ERROR' THEN DELETE;
 MSE=SS/DF;
 KEEP MSB DF;
DATA C2;
 MERGE Y X;
 IF TRT"=1 THEN DELETE;
 MEANPCT=MEANSURV/M;
DATA C3;
 MERGE Cl C2;
 TALPHA=TINV(.9 5,DF);
 LABEL M='NO. OF ORGANISMS AT START OF TEST'
       N='NO. OF REPLICATES'
       MEANPCT='MEAN REFERENCE SURVIVAL'
       MSE='MEAN SQUARE ERROR'
       DF='DEGREES OF FREEDOM, DF'
       TALPHA='T VALUE FOR  (1-ALPHA=0.95,DF)';
 TITLE2 'POWER OF LSD TO DETECT A TRUE POPULATION DIFFERENCE (D)';
 TITLE3 'FROM MEAN REFERENCE SURVIVAL USING ARCSINE TRANSFORMATION';
PROC PRINT LABEL NOOBS; VAR M N MEANPCT MSB DF TALPHA;
DATA C;
 SET C3;
 DO PCTDIFF=10 TO 50 BY 10;
  SEDSURV=MEANPCT-PCTDIFF/100;
  ARCSURV=ARSIN(SQRT{SEDSURV));
  ARCDIFF=MEAN-ARCSURV;
  TBETA=ARCDIFF*SQRT(N/(2*MSE))-TALPHA;
                                                                   DRAFT

-------
                                        D-63
  POWER=PROBT(TBETA,DF);
  OUTPUT;
  END;
 LABEL PCTDIFF='% REDUCTION IN SURVIVAL FROM REFERENCE'

       SEDSURV='DREDGED SEDIMENT SURVIVAL'
       ARCSURV='ARCSINE DREDGED SEDIMENT SURVIVAL'
       ARCDIFF='D'
       TBETA='T VALUE FOR (1-BETA,DF)';
PROC PRINT LABEL;
 VAR PCTDIFF SEDSURV ARCSURV ARCDIFF TBETA POWER;
 TITLE;
D4.2.2
BENTOX.SAS Program Output

OBS
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
TREATMENT
GROUP R
REFERENCE
REFERENCE
REFERENCE
REFERENCE
REFERENCE
SEDIMENT 1
SEDIMENT 1
SEDIMENT 1
SEDIMENT 1
SEDIMENT 1
SEDIMENT 2
SEDIMENT 2
SEDIMENT 2
SEDIMENT 2
SEDIMENT 2
SEDIMENT 3
SEDIMENT 3
SEDIMENT 3
SEDIMENT 3
SEDIMENT 3

EPLI'
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
                             BENTHIC TOXICITY DATA

                               NO. OF
                             ORGANISMS    NUMBER
                                PER         OF
                  REPLICATE  REPLICATE  SURVIVORS
                                 20
                                 20
                                 20
                                 20
                                 20
                                 20
                                 20
                                 20
                                 20
                                 20
                                 20
                                 20
                                 20
                                 20
                                 20
                                 20
                                 20
                                 20
                                 20
                                 20
                                20
                                20
                                19
                                19
                                20
                                17
                                16
                                18
                                17
                                15
                                15
                                16
                                13
                                17
                                11
                                17
                                12
                                10
                                16
                                13

ARCSINE
TRANSFORMATION
1.57080
1.57080
1.34528
1.34528
1.57080
1.17310
1.10715
1.24905
1.17310
1.04720
1.04720
1.10715
0.93774
1.17310
0.83548
1.17310
0.88608
0.78540
1.10715
0.93774
NORMALIZED
RANK FOR
SURVIVAL
1.46660
1.46660
0.83164
0.83164
1.46660
0.25276
-0.18775
0.58946
0.25276
-0.51861
-0.51861
-0.18775
-0.83164
0.25276
-1.40341
0.25276
-1.12814
-1.86824
-0.18775
-0.83164
                                BENTHIC TOXICITY DATA

OBS
1
2
3
4
TREATMENT
GROUP
REFERENCE
SEDIMENT 1
SEDIMENT 2
SEDIMENT 3

N
5
5
5
5

TOTAL
98
83
72
68
MEAN
SURVIVAL
19.6
16.6
14.4
13.6

SE
0.24495
0.50990
1.07703
1.28841
                                                                  DRAFT

-------
                                        D-64
                                BENTHIC  TOXICITY  DATA
                         ARCSINE-SQUARE ROOT TRANSFORMATION
Variable=RESID
                                BENTHIC TOXICITY  DATA
                         ARCSINE-SQUARE ROOT TRANSFORMATION
                          SHAPIRO-WILKS TEST FOR NORMALITY
                                UNIVARIATE PROCEDURE
                   N
                   W:Normal
      20
0.945932
Prob F

Model                    3       0.01373434      0.00457811     1.74    0.1985

Error

Corrected Total

                                BENTHIC TOXICITY  DATA
                         ARCSINE-SQUARE ROOT TRANSFORMATION
                                      LSD TEST

                           General  Linear Models  Procedure

                         T tests (LSD)  for variable:  ARCSURV

            NOTE: This  test  controls the type  I comparisonwise error  rate  not the
                  experimentwise error rate.

                       Alpha= 0.1   df= 16  MSE=  0.016175
                           Critical Value of T=  1.75
                     Least Significant Difference= 0.1404

          Means with the same letter are not significantly different.
T Grouping
A
B
B
C B
C
C

1
1

1

0
Mean
.4806
.1499

.0201

.9779
N
5
5

5

5
TRT
REFERENCE
SEDIMENT

SEDIMENT

SEDIMENT


1

2

3
                                                                   DRAFT

-------
Variable: ARCSURV

       TRT       N
                 D-65

         BENTHIC TOXICITY DATA
  ARCSINE-SQUARE ROOT TRANSFORMATION
                T-TEST

            TTEST PROCEDURE

ARCSINE TRANSFORMATION

           Mean             Std Dev
                      Std Error
REFERENCE
SEDIMENT 1
Variances
Unequal
Equal

5
5
5
5
T
.0930
.0930
DF
6.7
8.0
1
1

.48059096 0.12351878
.14991717 0.07629145
Prob> I T j
0.0017
0.0009
0
0

.05523928
.03411857

For HO: Variances are equal, F' = 2.62
                   DF
(4,4)
Prob>F'
0.3733
Variable: ARCSURV

       TRT       N
         BENTHIC TOXICITY  DATA
  ARCSINE-SQUARE ROOT TRANSFORMATION
                T-TEST

            TTEST PROCEDURE

ARCSINE TRANSFORMATION

           Mean             Std Dev
                      Std Error
REFERENCE
SEDIMENT 2
Variances
Unequal
Equal

5
5
5
5
T
.6335
.6335
DF
7.9
8.0
1.48059096 0.12351878
1.02013391 0.13470903
Prob> | T j
0.0005
0.0005
0.
0.

05523928
06024371

For HO: Variances are equal, F' =1.19    DF = (4,4)
                                 Prob>F' = 0.8706
Variable: ARCSURV

       TRT       N
         BENTHIC  TOXICITY  DATA
  ARCSINE-SQUARE ROOT TRANSFORMATION
                T-TEST

            TTEST PROCEDURE

ARCSINE TRANSFORMATION

           Mean             Std Dev
                      Std Error
REFERENCE
SEDIMENT 3
Variances
Unequal
Equal
5
5
T
5.5695
5.5695
DF
7.5
8.0
1.48059096 0.12351878
0.97789308 0.15961511
Prob>|T|
0.0007
0.0005
0.
0.

05523928
07138205

For HO: Variances are equal, F' = 1.67    DF = (4,4)    Prob>F' = 0.6315
                                                                  DRAFT

-------
                                        D-66

                                BENTHIC TOXICITY  DATA
                         SURVIVAL DATA CONVERTED TO RANKITS
                          SHAPIRO-WILKS TEST FOR NORMALITY

                                UNIVARIATE PROCEDURE
Variable=RESID
                   N                20
                   W:Normal   0.981773  Prob F

Model                    3       0.31609842      0.10536614     1.18    0.3493
Error                   16       1.43149144      0.08946821

Corrected Total         19       1.74758986

                                BENTHIC TOXICITY  DATA
                         SURVIVAL DATA CONVERTED TO RANKITS
                                 LSD TEST ON RANKITS

                           General  Linear Models  Procedure

                         T tests (LSD) for variable: RANKIT

            NOTE: This test  controls  the type I comparisonwise error  rate not  t
                  experimentwise error rate.

                       Alpha= 0.1  df= 16  MSE= 0.346143
                           Critical Value of T= 1.75
                     Least Significant Difference= 0.6496

             Means with the same letter are not significantly different.


                   T Grouping              Mean      N  TRT

                            A             1.213      5  REFERENCE

                            B             0.078      5  SEDIMENT 1
                            B
                    C       B            -0.538      5  SEDIMENT 2
                    C
                    C                    -0.753      5  SEDIMENT 3
                                                                   DRAFT

-------
                                        D-67
Variable: RANKIT

       TRT       N
         BENTHIC  TOXICITY  DATA
  SURVIVAL DATA CONVERTED TO RANKITS
           T-TEST ON RANKITS

            TTEST PROCEDURE

RANK FOR VARIABLE SURV

           Mean             Std Dev
                  Std Error
REFERENCE
SEDIMENT 1
Variances
Unequal
Equal

4
4
5
5
T
.5707
.5707
DF
7.6
8.0
1.21261524 0.34778201
0.07772091 0.43279236
Prob> j T j
0.0021
0.0018
0.15553284
0.19355063

For HO: Variances are equal, F' = 1.55    DF = (4,4)
                                 Prob>F'
               0.6821
Variable: RANKIT

       TRT       N
         BENTHIC  TOXICITY  DATA
  SURVIVAL DATA CONVERTED TO RANKITS
           T-TEST ON RANKITS

            TTEST PROCEDURE
RANK FOR VARIABLE SURV
           Mean
Std Dev
Std Error
REFERENCE
SEDIMENT 2
Variances
Unequal
Equal
5
5
T
5.4442
5.4442
1
-0
DF
6.2
8.0
.21261524 0.34778201
.53773198 0.62918751
Prob> | T j
0.0014
0.0006
0.15553284
0.28138121

For HO: Variances are equal, F' = 3.27    DF = (4,4)    Prob>F' = 0.2773

                                BENTHIC TOXICITY  DATA
                         SURVIVAL DATA CONVERTED TO RANKITS
                                  T-TEST ON  RANKITS
Variable: RANKIT

       TRT       N
            TTEST  PROCEDURE

RANK FOR VARIABLE SURV

           Mean             Std Dev
                  Std Error
REFERENCE
SEDIMENT 3
Variances
Unequal
Equal

4
4
5
5
T
.9088
.9088
1.
-0.
DF
5.4
8.0
21261524 0.34778201
75260418 0.82488344
Prob> [ T j
0.0038
0.0012
0.15553284
0.36889909

For HO: Variances are equal, F' = 5.63    DF = (4,4)
                                 Prob>F' = 0.1229
                                                                   DRAFT

-------
                                            D-68
                                   BENTHIC TOXICITY DATA
                POWER  OF LSD TO DETECT A TRUE POPULATION DIFFERENCE (D)
               FROM MEAN REFERENCE SURVIVAL  USING ARCSINE TRANSFORMATION
NO. OF
ORGANISMS
AT START
OF TEST
NO. OF
REPLICATES
MEAN
REFERENCE
SURVIVAL
MEAN
SQUARE
ERROR
DEGREES
OF
FREEDOM,
DF
      20            5

         % REDUCTION
             IN
          SURVIVAL
            FROM
 OBS     REFERENCE

  1          10
  2          20
  3          30
  4          40
  5          50
                   0.98
0.016175
16
   T VALUE FOR
(1-ALPHA=0.95,DF)

     1.74588
HEDGED
DIMENT
'RVIVAL
0.88
0.78
0.68
0.58
0.48
ARCSINE
DREDGED
SEDIMENT
SURVIVAL
1.21705
1.08259
0.96953
0.86574
0.76539


D
0.26354
0.39800
0.51106
0.61485
0.71520
T VALUE
FOR
(1-BETA,DF)
1.53043
3.20210
4.60766
5.89797
7.14555


POWER
0.92728
0.99722
0.99985
0.99999
1.00000
D4.3
Program BIOACC.SAS for Single-Time Point Bioaccumulation Test Data Analysis
BIOACC.SAS is a program to  compare Tier III bioaccumulation data from dredged sediments vs. reference
sediment, using raw data and logto transformation. Included in these analyses are: mean bioaccumulation from each
sediment exposure, Shapiro-Wilk's Test for normality, Levene's Test for equality of variances, /-tests for equal
unequal variances, LSD test, and tests on rankits (normalized ranks for contaminant concentration).  Refer to the
decision tree in Figures D-5A and 5B to determine which test results should be used. The program includes power
calculations for an LSD test on untransformed bioaccumulation data.
D4.3.1
BIOACC.SAS Program Statements
LIBNAME Q  'C:\SAS';
OPTIONS LINESIZE=79 PAGESIZE=59 NODATE NONUMBER;

/*  Identify the treatment codes.  */

PROC FORMAT;
 VALUE TRTFMT
    1='REFERENCE '
    2='SEDIMENT 1'
    3='SEDIMENT 2'
    4='SEDIMENT 3';

/*  Input the bioaccumulation data after the  CARDS statement, listing the  */
/*  treatment code, replicate, and contaminant  concentration.  A permanent  */
/*  SAS data set is created in the directory  specified  in the LIBNAME */
/*  statement.  */
                                                                         DRAFT

-------
                                        D-69

DATA Q.BIOACC;
 INPUT TRT REP CONC @@;
 CARDS;
1 1 .06 1 2 .05 1 3 .05 1 4 .08 1 5 .09
2 1 .16 2 2 .19 2 3 .18 2 4 .22 2 5 .31
31 .24 3 2 .10 3 3 .13 3 4 .18 3 5 .30
4 1 .13 4 2 .05 4 3 .17 4 4 .08 4 5 .22
.
i

I* Format, print, sort the data.  Print no. of observations, mean, and */
/* standard error for concentration in each treatment for both */
/* untransformed and loglO-transformed data.  Calculate rankits. */

DATA AO;
 SET Q.BIOACC;
 LOGCONC=LOG10(CONC);
 MERGEVAR=1;
 LABEL TRT='TREATMENT GROUP'
       REP='REPLICATE'
       CONC='CONTAMINANT CONCENTRATION, ug/g'
       LOGCONC='LOGIC CONCENTRATION';
 FORMAT TRT TRTFMT.;
 TITLE 'SINGLE-TIME POINT CONTAMINANT BIOACCUMULATION DATA';
PROC RANK NORMAL=BLOM OUT=A;
 VAR CONC; RANKS RANKIT;
PROC PRINT LABEL; VAR TRT REP CONC LOGCONC RANKIT;
 LABEL RANKIT='NORMALIZED RANK FOR CONCENTRATION';
PROC SORT; BY TRT;
PROC MEANS NOPRINT; BY TRT; VAR CONC LOGCONC; ID MERGEVAR;
 OUTPUT OUT=Y N=N NLOG MEAN=MEANCONC MEANLOG VAR=S2 S2LOG STDERR=SE SELOG;
PROC PRINT LABEL; VAR TRT N MEANCONC S2 SE MEANLOG S2LOG SELOG;
 LABEL    MEANCONC='MEAN CONTAMINANT CONC.'
      S2='VARIANCE'
      SE='STANDARD ERROR'
      MEANLOG='MEAN LOG10 CONC.'
      S2LOG='VARIANCE OF LOGS'
      SELOG='STANDARD ERROR OF LOGS';

/* Test normality of residuals of untransformed and log-transformed data */
/* using Shapiro-Wilk's Test. */

PROC GLM NOPRINT DATA=A;
 CLASS TRT;
 MODEL CONC LOGCONC=TRT;
 OUTPUT OUT=Z R=RESID RESIDLOG;
PROC UNIVARIATE NORMAL;
 VAR RESID RESIDLOG;
 TITLE2 'SHAPIRO-WILKS TEST FOR NORMALITY';

/* Conduct Levene's Test for equality of variances of untransformed and */
/* log-transformed data. */

DATA AY;
 MERGE AY; BY TRT;
 ABSDEV=ABS(CONC-MEANCONC);
 ABSLOG=ABS(LOGCONC-MEANLOG);
 LABEL    ABSDEV='ABSOLUTE DEVIATIONS FROM MEAN CONC.'
      ABSLOG='ABSOLUTE DEVIATIONS FROM MEAN LOGCONC.';
PROC GLM;
 CLASS TRT;
 MODEL ABSDEV ABSLOG=TRT;
 TITLE2 'LEVENE"S TEST';
                                                                   DRAFT

-------
                                        D-70

/* Perform LSD on untransformed and log-transformed data. */

PROC GLM DATA=A OUTSTAT=W1;
 CLASS TRT;
 MODEL CONC=TRT;
 MEANS TRT/LSD ALPHA=.l;
 TITLE2 'LSD TEST (UNTRANSFORMED DATA)';
PROC GLM DATA=A OUTSTAT=W2;
 CLASS TRT;
 MODEL LOGCONC=TRT;
 MEANS TRT/LSD ALPHA=.l;
 TITLE2 'LSD TEST (LOG-TRANSFORMED DATA)';

/* Perform t-tests for each dredged sediment-reference sediment comparison */
/* using untransformed and log-transformed data. */

DATA Tl;
 SET A;
 IF TRT>2 THEN DELETE;
PROC TTEST;
 CLASS TRT;
 VAR CONC LOGCONC;
 TITLE2 'T-TEST';
DATA T2;
 SET A;
 IF TRT=2 OR TRT=4 THEN DELETE;
PROC TTEST;
 CLASS TRT;
 VAR CONC LOGCONC;
DATA T3;
 SET A;
 IF TRT=2 OR TRT=3 THEN DELETE;
PROC TTEST;
 CLASS TRT;
 VAR CONC LOGCONC;

/* Test normality and equality of variances of rankits. */

PROC GLM NOPRINT DATA=A;
 CLASS TRT;
 MODEL RANKIT=TRT;
 OUTPUT OUT=Z2 R=RESID;
 TITLE2 'BIOACCUMULATION DATA CONVERTED TO RANKITS';
PROC UNIVARIATE NORMAL;
 VAR RESID;
 TITLE3 'SHAPIRO-WILKS TEST FOR NORMALITY';
PROC MEANS DATA=A NOPRINT;
 BY TRT; VAR RANKIT;
 OUTPUT OUT=X MEAN=MEAN;
DATA AX;
 MERGE AX; BY TRT;
 ABSDEV=ABS(RANKIT-MEAN);
PROC GLM;
 CLASS TRT;
 MODEL ABSDEV=TRT;
 TITLES 'LEVENE"S TEST';

/* Perform LSD on rankits. */

PROC GLM DATA=A;
 CLASS TRT;
 MODEL RANKIT=TRT;
 MEANS TRT/LSD ALPHA=.l;
                                                                   DRAFT

-------
                                        D-71
 TITLES 'LSD TEST';
/* Perform t-tests for each dredged sediment-reference sediment comparison  */
/* using rankits. */

PROC TTEST DATA=T1;
 CLASS TRT;
 VAR RANKIT;
 TITLES 'T-TEST';
PROC TTEST DATA=T2;
 CLASS TRT;
 VAR RANKIT;
PROC TTEST DATA=T3;
 CLASS TRT;
 VAR RANKIT;

/* Calculate power of an LSD test to detect true population differences */
/* 10, 25, 50, and 100% above the reference mean contaminant concentration.  */

DATA Cl;
 SET Wl;
 IF _TYPE_A='ERROR' THEN DELETE;
 MSE=SS/DF;
 MERGEVAR=1;
 KEEP MSB DF MERGEVAR;
DATA C2;
 SET Y;
 IF TRT*=1 THEN DELETE;
DATA C3;
 MERGE Cl C2;
 TALPHA=TINV(.95,DF);
 LABEL    N='NO. OF REPLICATES, N'
      MEANCONC='REFERENCE MEAN CONTAMINANT CONCENTRATION'
      MSE='MEAN SQUARE ERROR, MSB'
      DF='DEGREES OF FREEDOM, DF'
      TALPHA='T VALUE FOR (1-ALPHA=0.95,DF)';
 TITLE2 'POWER OF LSD TO DETECT A TRUE POPULATION DIFFERENCE (D)';
 TITLE3 'ABOVE REFERENCE MEAN CONTAMINANT CONCENTRATION';
PROC PRINT LABEL NOOBS; VAR N MEANCONC MSB DF TALPHA;
DATA C4;
 SET C3;
 DO PCTDIFF=10,25,50,100,200,300;
  SEDCONC=MEANCONC+((PCTDIFF/100)*MEANCONC);
  D=SEDCONC-MEANCONC;
  TBETA=D*SQRT(N/(2*MSE))-TALPHA;
  POWER=PROBT(TBETA,DF);
  OUTPUT;
  END;
 LABEL    PCTDIFF='% INCREASE IN CONC. ABOVE REFERENCE'
      SEDCONC='DREDGED SEDIMENT BIOACCUMULATION'
      TBETA='T VALUE FOR (1-BETA,DF)'
      POWER='POWER (1-BETA)';
PROC PRINT LABEL NOOBS; VAR PCTDIFF SEDCONC D TBETA POWER;
 TITLE 'POWER OF LSD TO DETECT % INCREASE IN CONCENTRATION ABOVE REFERENCE';
 TITLE2 'MEAN CONTAMINANT CONCENTRATION GIVEN N, MSB AND DF SHOWN ABOVE';
DATA C5;
 SET C3;
 DO POWER=.5,.6,.7,.8,.9,.95,.99;
  TBETA=TINV(POWER,DF);
  D=((TBETA+TALPHA)*SQRT(2*MSE))/SQRT(N);
  SEDCONC=MEANCONC+D;
  PCTDIFF=(D*100)/MEANCONC;
  OUTPUT;
                                                                   DRAFT

-------
                                        D-72

  END;
 LABEL    SEDCONC='DREDGED SEDIMENT BIOACCUMULATION'
      PCTDIFF='% INCREASE IN CONC. ABOVE REFERENCE'
      TBETA='T VALUE FOR (1-BETA,DF)'
      POWER='POWER (1-BETA)';
PROC PRINT LABEL NOOBS; VAR POWER D SEDCONC PCTDIFF TBETA;
 TITLE 'MINIMUM DREDGED SEDIMENT BIOACCUMULATION THAT CAN BE DETECTED BY LSD';
 TITLE2 'AS SIGNIFICANT GIVEN SPECIFIED POWER AND N, MSB, AND DF SHOWN ABOVE';

/* Calculation of upper confidence limits (UCL) for comparison of mean */
/* dredged sediment bioaccumulation with an action level. */

DATA D;
 MERGE Cl Y; BY MERGEVAR;
 IF TRT=1 THEN DELETE;
 TALPHA1=TINV(.95,DF);
 TALPHA2=TINV(.95,N-1);
 UCL1=MEANCONC+TALPHA1*(SQRT(MSE/N) ) ;
 UCL2=MEANCONC+TALPHA2 *(SQRT(S2/N));
 DMIN1=TALPHA1*SQRT(MSE/N);
 DMIN2=TALPHA2 *SQRT(S2/N);
 LABEL    UCL1='UCL (EQUAL VARIANCES)'
      UCL2='UCL (UNEQUAL VARIANCES)'
      TALPHA1='T VALUE FOR (1-ALPHA=.95,DF)'
      TALPHA2 ='T VALUE FOR (1-ALPHA=.9 5,N-1)'
      DMIN1='MINIMUM SIGNIFICANT DIFFERENCE'
      DMIN2='MINIMUM SIGNIFICANT DIFFERENCE'
      MSE='MEAN SQUARE ERROR'
      S2='VARIANCE'
      MEANCONC='MEAN BIOACCUMULATION';
 TITLE 'COMPARISON OF MEAN DREDGED SEDIMENT BIOACCUMULATION WITH ACTION
LEVEL:';
PROC PRINT LABEL NOOBS; VAR TRT MEANCONC UCL1 MSB TALPHA1 DF DMIN1;
 TITLE2 'UPPER CONFIDENCE LIMITS (UCL) WHEN VARIANCES ARE EQUAL';
PROC PRINT LABEL NOOBS; VAR TRT MEANCONC UCL2 S2 TALPHA2 N DMIN2;
 TITLE2 'UPPER CONFIDENCE LIMITS (UCL) WHEN VARIANCES ARE UNEQUAL';


/* Calculate power of dredged sediment-action level comparisons using */
/* MSB given 10, 20, 30, 40, and 50% decreases in mean concentration */
/* below action level. */

DATA Dl;
 SET C3;
 ACTION=.2;
 DO PCTDIFF=10 TO 50 BY 10;
  D=PCTDIFF*ACTION/100;
  SEDCONC=ACTION-D;
  TBETA=D*SQRT(N/MSE)-TALPHA;
  POWER=PROBT(TBETA,DF);
  OUTPUT;
  END;
 LABEL    PCTDIFF='% DECREASE BELOW ACTION LEVEL'
      SEDCONC='MEAN DREDGED SEDIMENT BIOACCUMULATION'
      TBETA='T VALUE FOR  (1-BETA,DF)'
      POWER='POWER (1-BETA)';
PROC PRINT NOOBS LABEL; VAR PCTDIFF SEDCONC D TBETA POWER;
 TITLE 'POWER TO DETECT % DECREASE IN CONCENTRATION BELOW';
 TITLE2 'ACTION LEVEL OF 0.2 ug/g GIVEN N, MSB AND DF SHOWN ABOVE';
                                                                   DRAFT

-------
                                        D-73
D4.3.2
BIOACC.SAS Program Output

     SINGLE-TIME POINT CONTAMINANT BIOACCUMULATION DATA
                                  CONTAMINANT
                                                                   NORMALIZED

IBS
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
TREATMENT
GROUP
REFERENCE
REFERENCE
REFERENCE
REFERENCE
REFERENCE
SEDIMENT 1
SEDIMENT 1
SEDIMENT 1
SEDIMENT 1
SEDIMENT 1
SEDIMENT 2
SEDIMENT 2
SEDIMENT 2
SEDIMENT 2
SEDIMENT 2
SEDIMENT 3
SEDIMENT 3
SEDIMENT 3
SEDIMENT 3
SEDIMENT 3
CONCENTRAT ION , LOG 1 0
REPLICATE
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
ug/g
0.06
0.05
0.05
0.08
0.09
0.16
0.19
0.18
0.22
0.31
0.24
0.10
0.13
0.18
0.30
0.13
0.05
0.17
0.08
0.22
CONCENTRATION CO»
-1.22185
-1.30103
-1.30103
-1.09691
-1.04576
-0.79588
-0.72125
-0.74473
-0.65758
-0.50864
-0.61979
-1.00000
-0.88606
-0.74473
-0.52288
-0.88606
-1.30103
-0.76955
-1.09691
-0.65758
RANK FOR
ICENTRATK
-0.91914
-1.46660
-1.46660
-0.66680
-0.44777
0.06193
0.58946
0.38117
0.83164
1.86824
1.12814
-0.31457
-0.12434
0.38117
1.40341
-0.12434
-1.46660
0.18676
-0.66680
0.83164
SINGLE-TIME POINT CONTAMINANT BIOACCUMULATION DATA


OBS
1
2
3
4

TREATMENT
GROUP
REFERENCE
SEDIMENT 1
SEDIMENT 2
SEDIMENT 3
MEAN
CONTAMINANT
N CONC.
5 0.066
5 0.212
5 0.190
5 0.130


VARIANCE
.00033
.00347
.00660
.00465
MEAN
STANDARD LOG10 VARIANCE
ERROR CONC. OF LOGS
0.008124 -1.19332 0.013772
0.026344 -0.68561 0.012257
0.036332 -0.75469 0.037367
0.030496 -0.94223 0.066666
STANDARD
ERROR OF
LOGS
0.05248
0.04951
0.08645
0.11547
Variable=RESID
     SINGLE-TIME POINT CONTAMINANT BIOACCUMULATION DATA
              SHAPIRO-WILKS TEST FOR NORMALITY

                    UNIVARIATE PROCEDURE

       N                20
       W:Normal   0.957973  Prob
-------
                            D-74

     SINGLE-TIME POINT CONTAMINANT BIOACCUMULATION DATA
                       LEVENE'S TEST
              General Linear Models Procedure
Dependent Variable:
Source
Model
Error
Corrected Total
Dependent Variable:
Source
Model
Error
Corrected Total
ABSDEV
DF
3
16
19
ABSLOG
DF
3
16
19
ABSOLUTE DEVIATIONS
Sum of
Squares
0.00647280
0.01605600
0.02252880
ABSOLUTE DEVIATIONS
Sum of
Squares
0.04702396
0.11456390
0.16158786
FROM MEAN CONC.
Mean
Square F Value
0.00215760 2.15
0.00100350

FROM MEAN LOGCONC.
Mean
Square F Value
0.01567465 2.19
0.00716024

Pr > F
0.1339


Pr > F
0.1291


     SINGLE-TIME POINT CONTAMINANT BIOACCUMULATION DATA
                LSD TEST  (UNTRANSFORMED DATA)

              General Linear Models Procedure

              T tests (LSD) for variable: CONC

NOTE: This test  controls the type I  comparisonwise error rate  not
      experimentwise error rate.


             Alpha=  0.1   df= 16 MSE= 0.003763
                  Critical Value of T= 1.75
            Least Significant Difference= 0.0677

 Means  with the  same  letter are  not significantly  different.
       T Grouping
        B
        B
        B
A
A
A

C
C
C
  Mean      N  TRT

0.2120      5  SEDIMENT 1

0.1900      5  SEDIMENT 2

0.1300      5  SEDIMENT 3

0.0660      5  REFERENCE
                                                       DRAFT

-------
                                        D-75

                           LSD TEST (LOG-TRANSFORMED  DATA)

                          Alpha= 0.1  df=  16  MSE= 0.032515
                              Critical Value of  T= 1.75
                        Least Significant Difference= 0.1991

             Means  with the same letter are  not  significantly different.
                   T Grouping
                    B
                    B
                    B
                 A
                 A
                 A
                   Mean      N  TRT

                 -0.686      5  SEDIMENT 1

                 -0.755      5  SEDIMENT 2

                 -0.942      5  SEDIMENT 3

                 -1.193      5  REFERENCE
Variable: CONC
      SINGLE-TIME POINT CONTAMINANT BIOACCUMULATION DATA
                            T-TEST

                        TTEST  PROCEDURE

            CONTAMINANT CONCENTRATION, ug/g
       TRT
      N
          Mean
                                  Std Dev
Std Error
REFERENCE
SEDIMENT 1
Variances
Unequal
Equal
5
5
T
-5.2960
-5.2960
DF
4.8
8.0
0.
0.

06600000 0.01816590
21200000 0.05890671
Prot»|Tj
0.0039
0.0007
0.00812404
0.02634388

For HO: Variances are equal, F'
                        10.52
                    DF = (4,4)
                                         Prot»F' = 0.0426
Variable: LOGCONC

       TRT       N
            LOG10 CONCENTRATION

                       Mean
                           Std Dev
                                                    Std Error
 REFERENCE
SEDIMENT 1

Variances
5         -1.19331525
5         -0.68561391

T       DF    Prob>|T|
Unequal
Equal
-7.0366
-7.0366
8.0
8.0
                0.0001
                0.0001
For HO: Variances are equal, F' = 1.12
                                     0.11735241
                                     0.11071260
                                            0.05248159
                                            0.04951218
                               DF = (4,4)
                                Prob>F' = 0.9128
                                                                   DRAFT

-------
                                        D-76
Variable: CONC

       TRT
      SINGLE-TIME POINT CONTAMINANT BIOACCUMULATION DATA
                            T-TEST

                       TTEST PROCEDURE

            CONTAMINANT CONCENTRATION,  ug/g

      N                Mean             Std Dev
                                             Std Error
 REFERENCE
SEDIMENT 2

Variances
      5          0.06600000
      5          0.19000000

      T       DF    Prob> j T j
Unequal
Equal
-3.3307
-3.3307
4.4
8.0
0.0258
0.0104
For HO: Variances are equal, F' =  20.00
                        0.01816590
                        0.08124038
                                   0.00812404
                                   0.03633180
                                 DF
                         (4,4)
                         Prob>F'
0.0132
Variable: LOGCONC

       TRT       N
            LOGIC CONCENTRATION

                       Mean
                           Std Dev
                                    Std Error
REFERENCE
SEDIMENT 2
Variances
Unequal
Equal
5
5
T
-4.3371
-4.3371
-1.
-0.
DF
6.6
8.0
19331525 0.11735241
75469033 0.19330562
Prob> j T j
0.0040
0.0025
0.05248159
0.08644890

For HO: Variances are equal, F' =2.71    DF = (4,4)
                                             Prob>F' = 0.3570
Variable: CONC

       TRT
      SINGLE-TIME POINT CONTAMINANT BIOACCUMULATION DATA
                            T-TEST

                        TTEST  PROCEDURE

            CONTAMINANT CONCENTRATION, ug/g

      N                Mean             Std Dev
                                             Std Error
REFERENCE
SEDIMENT 3
Variances
Unequal
Equal
5
5
T
-2.0279
-2.0279
DF
4.6
8.0
0.06600000 0.01816590
0.13000000 0.06819091
Prob>jTJ
0.1045
0.0771
0
0

.00812404
.03049590

For HO: Variances are equal, F'
                        14.09
                    DF = (4,4)
                         Prob>F'
0.0252
Variable: LOGCONC
            LOGIC CONCENTRATION
                                                                   DRAFT

-------
                                        D-77
       TRT
N
Mean
Std Dev
                                                                     Std Error
REFERENCE
SEDIMENT 3
Variances
Unequal
Equal

-1
-1
5
5
T
.9796
.9796
-1
-0
DF
5.6
8.0
.19331525 0.11735241
.94222501 0.25819757
Prob> j T I
0.0990
0.0831
0
0

.05248159
.11546947

For HO: Variances are equal, F' = 4.84
                         DF
             (4,4)
     Prot»F'
0.1558
Variable=RESID
                 SINGLE-TIME POINT CONTAMINANT BIOACCUMULATION DATA
                      BIOACCUMULATION DATA CONVERTED  TO RANKITS
                          SHAPIRO-WILKS TEST FOR NORMALITY
                   N
                   W:Normal
                                UNIVARIATE PROCEDURE
                   20
             0.972308  Prob F

      0.6212
                 SINGLE-TIME POINT CONTAMINANT BIOACCUMULATION DATA
                      BIOACCUMULATION DATA CONVERTED  TO  RANKITS
                                      LSD TEST

                           General  Linear  Models  Procedure

                         T tests (LSD) for variable:  RANKIT

            NOTE: This test controls the type I comparisonwise error rate not
                  the experimentwise error rate.
                                                                  DRAFT

-------
                                        D-78
                         Alpha=  0.1   df=  16  MSE=  0.503649
                              Critical Value of T=  1.75
                        Least Significant Difference= 0.7836

            Means with  the  same  letter  are not significantly different.
                   T Grouping
                    B
                    B
                    B
     A
     A
     A

     C
     C
     C
  Mean

 0.746

 0.495

-0.248

-0.993
N  TRT

5  SEDIMENT 1

5  SEDIMENT 2

5  SEDIMENT 3

5  REFERENCE
                 SINGLE-TIME POINT CONTAMINANT BIOACCUMULATION DATA
                      BIOACCUMULATION DATA CONVERTED  TO  RANKITS
Variable: RANKIT

       TRT       N
           TTEST PROCEDURE

RANK FOR VARIABLE CONC

           Mean             Std Dev
                            Std Error
REFERENCE
SEDIMENT 1
Variances
Unequal
Equal
5
5
T
-4.6920
-4.6920
-0
0
DF
7.0
8.0
.99338019 0.46306944
.74648762 0.68780736
Prob> j T j
0.0022
0.0016
0
0

.20709095
.30759680

For HO: Variances are equal, F' =2.21    DF= (4,4)
                                 Prob>F' = 0.4623
                 SINGLE-TIME POINT CONTAMINANT BIOACCUMULATION DATA
                      BIOACCUMULATION DATA CONVERTED  TO RANKITS
Variable: RANKIT

       TRT       N
            TTEST  PROCEDURE

RANK FOR VARIABLE CONC

           Mean
          Std Dev
                Std Error
REFERENCE
SEDIMENT 2
Variances
Unequal
Equal

-3
-3
5
5
T
.7583
.7583
-0
0
DF
6.6
8.0
.99338019 0.46306944
.49476200 0.75465812
Prob> j T j
0.0079
0.0056
0.20709095
0.33749337

For HO: Variances are equal, F' = 2.66    DF = (4,4)    Prob>F'
                                           0.3671
                                                                   DRAFT

-------
                                        D-79
                 SINGLE-TIME POINT CONTAMINANT BIOACCUMULATION DATA
                      BIOACCUMULATION DATA CONVERTED TO RANKITS
Variable: RANKIT

       TRT       N
                 TTEST  PROCEDURE

     RANK FOR VARIABLE CONC

                Mean             Std Dev
                                        Std Error
REFERENCE
SEDIMENT 3
5
5
-0.99338019
-0.24786944
0.46306944
0.87038805
0.20709095
0.38924937
Variances
       DF
  Prob>jT|
Unequal
Equal
-1.6908
-1.6908
6.1
8.0
0.1411
0.1293
For HO: Variances are equal, F' = 3.53
                        DF
                  (4,4)
         Prob>F'
       0.2491
                 SINGLE-TIME POINT CONTAMINANT BIOACCUMULATION DATA
               POWER OF LSD TO DETECT A TRUE POPULATION DIFFERENCE (D)
                   ABOVE REFERENCE MEAN CONTAMINANT CONCENTRATION
      NO. OF
   REPLICATES,
        N
  REFERENCE
     MEAN
 CONTAMINANT
CONCENTRATION

    0.066
        MEAN
       SQUARE
       ERROR,
         MSE

      .0037625
 DEGREES
   OF
FREEDOM,
   DF

   16
   T VALUE FOR
(1-ALPHA=0.95,DF)

     1.74588
         POWER OF LSD TO DETECT % INCREASE IN CONCENTRATION ABOVE REFERENCE
           MEAN CONTAMINANT CONCENTRATION GIVEN N, MSE AND DF SHOWN ABOVE

       % INCREASE
        IN CONC.        DREDGED                    T VALUE
         ABOVE          SEDIMENT                     FOR          POWER
       REFERENCE    BIOACCUMULATION       D       (1-BETA,DF)     (1-BETA)

           10            0.0726        0.0066      -1.57576      0.06732
           25            0.0825        0.0165      -1.32056      0.10261
           50            0.0990        0.0330      -0.89524      0.19196
          100            0.1320        0.0660      -0.04460      0.48249
          200            0.1980        0.1320       1.65668      0.94147
          300            0.2640        0.1980       3.35796      0.99800
        MINIMUM DREDGED SEDIMENT BIOACCUMULATION THAT CAN BE DETECTED BY LSD
         AS  SIGNIFICANT GIVEN SPECIFIED POWER AND  N,  MSE,  AND DF SHOWN ABOVE
        POWER
      (1-BETA)

        0.50
        0.60
        0.70
        0.80
        0.90
        0.95
        0.99
0.06773
0.07772
0.08849
0.10127
0.11959
0.13546
0.16796
    DREDGED
    SEDIMENT
BIOACCUMULATION

    0.13373
    0.14372
    0.15449
    0.16727
    0.18559
    0.20146
    0.23396
 % INCREASE
  IN CONC.
   ABOVE
 REFERENCE

  102.622
  117.763
  134.069
  153.446
  181.195
  205.244
  254.477
    T VALUE
      FOR
  (1-BETA,DF)

    0.00000
    0.25760
    0.53501
    0.86467
     .33676
     .74588
1.
1.
    2.58349
                                                                   DRAFT

-------
                                            D-80

        COMPARISON OF MEAN DREDGED SEDIMENT BIOACCUMULATION WITH ACTION LEVEL:
                UPPER CONFIDENCE LIMITS  (UCL) WHEN  VARIANCES  ARE EQUAL
TREATMENT
GROUP
SEDIMENT 1
SEDIMENT 2
SEDIMENT 3
MEAN
BIOACCUMULATION
0.212
0.190
0.130
UCL
(EQUAL
VARIANCES )
0.25989
0.23789
0.17789
MEAN
SQUARE
ERROR
.0037625
.0037625
.0037625
T VALUE FOR
(1-ALPHA=.95,DF)
1.74588
1.74588
1.74588
DF
16
16
16
MINIMUM
SIGNIFICANT
DIFFERENCE
0.047893
0.047893
0.047893
        COMPARISON OF MEAN DREDGED SEDIMENT BIOACCUMULATION WITH ACTION LEVEL:
               UPPER CONFIDENCE LIMITS  (UCL) WHEN VARIANCES ARE UNEQUAL
TREATMENT
  GROUP

SEDIMENT 1
SEDIMENT 2
SEDIMENT 3
                      UCL                                      MINIMUM
      MEAN        (UNEQUAL                T VALUE FOR      SIGNIFICANT
BIOACCUMULATION VARIANCES) VARIANCE  (1-ALPHA=.95,N-1)  N  DIFFERENCE
     0.212
     0.190
     0.130
0.26816
0.26745
0.19501
.00347
.00660
.00465
2.13185
2.13185
2.13185
5
5
5
0.056161
0.077454
0.065013
                   POWER TO DETECT  % DECREASE  IN CONCENTRATION BELOW
               ACTION LEVEL  OF 0.2 ug/g GIVEN N, MSB AND DF  SHOWN ABOVE
        % DECREASE
           BELOW
          ACTION
           LEVEL

            10
            20
            30
            40
            50
            MEAN DREDGED
              SEDIMENT
          BIOACCUMULATION

                0.18
                0.16
                0.14
                0.12
                0.10


D
0.02
0.04
0.06
0.08
0.10
T VALUE
FOR
(1-BETA,DF)
-1.01680
-0.28772
0.44136
1.17045
1.89953

POWER
(1-BETA)
0.16219
0.38863
0.66757
0.87052
0.96216
D4.4
 Program BIOACCSS.SAS for Time-Sequenced Bioaccumulation Test Data Analysis
BIOACCSS.SAS is a program to compare Tier IV estimated steady-state bioaccumulation (C^) from dredged

sediments vs. reference sediment, using untransformed data and Iog10 transformation. Included are: data plots,

estimation of CM, mean CM from each sediment exposure, Shapiro-Wilk's test for normality, Levene's test for

equality of variances, LSD test, f-tests for equal or unequal variances, and tests on rankits (normalized ranks for

C.J.  Refer to the decision tree in Figures D-5A and 5B to determine which test results should be used.  The

program includes power calculations for an LSD test on untransformed Ca estimates.
                                                                         DRAFT

-------
                                        D-81

D4.3.1       BIOACCSS.SAS Program Statements


LIBNAME Q 'C:\SAS';
OPTIONS LINESIZE=79 PAGESIZE=59 NONUMBER NODATE;

/* Identify the treatment codes. */

PROC FORMAT;
 VALUE TRTFMT
   1='REFERENCE '
   2='SEDIMENT 1*
   3='SEDIMENT 2'
   4='SEDIMENT 3';

/* Input the bioaccumulation data after the CARDS statement, listing the */
/* day, replicate, treatment code, and contaminant concentration.  A */
/* permanent SAS data set is created in the directory specified in the */
/* LIBNAME statement. */


DATA Q.BIOACCSS;
 INPUT DAY REP TRT CONC @@;
 CARDS;
2
2
2
2
4
4
4
4
7
7
7
7
1
1
1
1
1
1
1
1
1
1
1
1
1
2
3
4
1
2
3
4
1
2
3
4
•
•
•
*
1
*
1
1
054
159
869
745
441
516
838
.316
687
881
.246
.583
2
2
2
2
4
4
4
4
7
7
7
7
2
2
2
2
2
2
2
2
2
2
2
2
1
2
3
4
1
2
3
4
1
2
3
4
.163 231 .391 241 .234 2 5
.292 232 .428 242 .558 2 5
.726 233 .394 243 1.232 2
1.703 234 2.045 244 1.855
.797 431 .203 441 .564 4 5
.158 432 .743 442 .324 4 5
.633 433 .452 443 .728 4 5
.930 434 2.141 444 1.150
.177 731 .862 741 .413 7 5
.317 732 .270 742 .562 7 5
.816 733 .897 743 1.639 7
2.715 734 1.016 744 2.221
1
2
5
2
1
2
3
4
1
2
5
7
•
•
3
5
1
5
•
3
5
034
256
.977
4 1.135
018
126
.314
4 1.621
029
603
.688
4 2.13
10 1 1 .037 10 2 1 .549 10 3 1 .884 10 4 1 .787 10 5 1 .294
10 1 2 .278 10 2 2 .485 10 3 2 .051 10 4 2 .909 10 5 2 .718
10 1 3 1.767 10 2 3 1.272 10 3 3 1.003 10 4 3 1.158 10 5 3 1.415
10 1 4 1.578 10 2 4 2.268 10 3 4 1.756 10 4 4 2.899 10 5 4 .890
18 1 1 .856 18 2 1 .598 18 3 1 .016 18 4 1 .806 18 5 1 .119
18 1 2 .904 18 2 2 1.300 18 3 2 .671 18 4 2 .934 18 5 2 1.173
18 1 3 1.631 18 2 3 1.877 18 3 3 1.487 18 4 3 1.216 18 5 3 1.280
18 1 4 2.822 18 2 4 2.607 18 3 4 3.414 18 4 4 1.319 18 5 4 1.866
28 1 1 .514 28 2 1 .839 28 3 1 .793 28 4 1 .899 28 5 1 .226
28 1 2 .172 28 2 2 1.049 28 3 2 .476 28 4 2 .712 28 5 2 1.245
28 1 3 1.178 28 2 3 1.721 28 3 3 1.366 28 4 3 1.513 28 5 3 1.843
28 1 4 1.295 28 2 4 2.964 28 3 4 2.109 28 4 4 2.820 28 5 4 3.325
;

/* Specify contaminant concentrations in the sediments.  Format, sort, */
/* and print the data. */

DATA AA;
 SET Q.BIOACCSS;
 SELECT (TRT);
  WHEN (1) CS=.45;
  WHEN (2) CS=4;
  WHEN (3) CS=33;
  WHEN (4) CS=44;
  OTHERWISE;
 END;
 LABEL TRT='TREATMENT GROUP'
                                                                   DRAFT

-------
                                        D-82

       REP='REPLICATE'
       CONC='CONC. IN TISSUE'
       CS='CONC. IN SEDIMENT';
 FORMAT TRT TRTFMT.;
 TITLE 'TIME-SEQUENCED BIOACCUMULATION';
PROC SORT; BY TRT REP;
PROC PRINT LABEL; BY TRT; VAR REP DAY CONC CS;

/* Plot the data by treatment group, identifying the replicates.  Plots */
/* may be sent to the screen using the first GOPTIONS statement, or to a */
/* printer using the second GOPTIONS statement.  Consult the SAS/GRAPH */
/* User's Guide (SAS Institute, Inc., 1988c) for appropriate device names */
/* and instructions for GACCESS=. */

*GOPTIONS DEVICE=VGA;
GOPTIONS DEVICE=HPLJ3P GACCESS='SASGASTD>LPT2:' VSIZE=6 IN HSIZE=6.5 IN
   VORIGIN=3 IN HORIGIN=0.3 IN;
PROC GPLOT UNIFORM; BY TRT;
 PLOT CONC*DAY=REP;

/* Perform nonlinear regressions on each treatment and replicate. */
/* If you wish to use a method other than DUD, include the following */
/* derivative statements after the MODEL statement:  DER.K1=CS/K2*(1-EX); */
/* and DER.K2=CS*(K1/K2)*(DAY*EX-(1-EX)/K2);.  Save regression parameters */
/* in a permanent SAS data set. */

PROC NLIN BEST=10 METHOD=DUD;
 BY TRT REP;
 FARMS Kl=0 TO 3 BY .1 K2=.01 TO 2 BY . 1;
 EX=EXP(-K2*DAY);
 MODEL CONC=CS*(K1/K2)*(1-EX);
 OUTPUT OUT=Q.REGPARMS PARMS=K1 K2;

/* Calculate and print Css and regression parameters.  Log-transform Css. */
/* Calculate rankits.  Save these variables in a permanent SAS data set. */

DATA A;
 SET Q.REGPARMS;
 IF DAY<28 THEN DELETE;
 CSS=CS*K1/K2;
 LOGCSS=LOG10(CSS);
 DROP DAY CONC;
 LABEL    CSS='STEADY STATE CONC., Css'
      LOGCSS='LoglO Css'
      Kl='UPTAKE RATE CONSTANT, kl'
      K2='DEPURATION RATE CONSTANT, k2';
 MERGEVAR=1;

PROC RANK NORMAL=BLOM OUT=Q.CSS;
 VAR CSS; RANKS RANKIT;
PROC PRINT LABEL DATA=Q.CSS; VAR TRT REP Kl K2 CSS LOGCSS RANKIT;
 LABEL      RANKIT='NORMALIZED RANK FOR Css';

/* Calculate and print descriptive statistics for Css and logCss. */

PROC MEANS NOPRINT DATA=Q.CSS; BY TRT; VAR CSS LOGCSS; ID MERGEVAR;
 OUTPUT OUT=Y N=N NLOG MEAN=MEANCSS MEANLOG VAR=S2 S2LOG STDERR=SE SELOG;
PROC PRINT LABEL; VAR TRT N MEANCSS S2 SE MEANLOG S2LOG SELOG;
 LABEL    MEANCSS='MEAN Css'
      S2='VARIANCE'
      SE='STANDARD ERROR'
      MEANLOG='MEAN LoglO Css'
      S2LOG='VARIANCE OF LOGS'
                                                                   DRAFT

-------
                                        D-83

      SELOG='STANDARD ERROR OF LOGS';

/* Test normality of residuals of untransformed and log-transformed Css */
/* using Shapiro-Wilk's Test. */

PROC GLM NOPRINT DATA=Q.CSS;
 CLASS TRT;
 MODEL CSS LOGCSS=TRT;
 OUTPUT OUT=Z R=RESID RESIDLOG;
PROC UNIVARIATE NORMAL;
 VAR RESID RESIDLOG;
 TITLE2 'SHAPIRO-WILKS TEST FOR NORMALITY';
/* Conduct Levene's Test for equality of variances of untransformed and */
/* log-transformed Css. */

DATA AX;
 MERGE Q.CSS Y; BY TRT;
 ABSDEV=ABS(CSS-MEANCSS);
 ABSLOG=ABS(LOGCSS-MEANLOG);
 LABEL    ABSDEV='ABSOLUTE DEVIATIONS FROM Css MEAN'
      ABSLOG='ABSOLUTE DEVIATIONS FROM logCss MEAN';
PROC GLM;
 CLASS TRT;
 MODEL ABSDEV ABSLOG=TRT;
 TITLE2 'LEVENE''S TEST';

/* Perform LSD on untransformed and log-transformed Css. */

PROC GLM DATA=Q.CSS OUTSTAT=W1;
 CLASS TRT;
 MODEL CSS=TRT;
 MEANS TRT/LSD ALPHA=.l;
 TITLE2 'LSD TEST (UNTRANSFORMED DATA)';
PROC GLM DATA=Q.CSS OUTSTAT=W2;
 CLASS TRT;
 MODEL LOGCSS=TRT;
 MEANS TRT/LSD ALPHA=.l;
 TITLE2 'LSD TEST (LOG-TRANSFORMED DATA)';

/* Perform t-tests for each dredged sediment-reference sediment comparison */
/* using untransformed and log-transformed Css. */

DATA Tl;
 SET Q.CSS;
 IF TRT>2 THEN DELETE;
PROC TTEST;
 CLASS TRT;
 VAR CSS LOGCSS;
 TITLE2 'T-TEST';
DATA T2;
 SET Q.CSS;
 IF TRT=2 OR TRT=4 THEN DELETE;
PROC TTEST;
 CLASS TRT;
 VAR CSS LOGCSS;
DATA T3;
 SET Q.CSS;
 IF TRT=2 OR TRT=3 THEN DELETE;
PROC TTEST;
 CLASS TRT;
 VAR CSS LOGCSS;

/* Test normality and equality of variances of rankits. */
                                                                   DRAFT

-------
                                        D-84

PROC GLM NOPRINT DATA=Q.CSS;
 CLASS TRT;
 MODEL RANKIT=TRT;
 OUTPUT OUT=Z1 R=RESID;
 TITLE2 'Css CONVERTED TO RANKITS';
PROC UNIVARIATE NORMAL;
 VAR RESID;
 TITLES 'SHAPIRO-WILKS TEST FOR NORMALITY';
PROC MEANS DATA=Q.CSS NOPRINT;
 BY TRT; VAR RANKIT;
 OUTPUT OUT=X2 MEAN=MEAN;
DATA AX2;
 MERGE Q.CSS X2; BY TRT;
 ABSDEV=ABS(RANKIT-MEAN);
PROC GLM;
 CLASS TRT;
 MODEL ABSDEV=TRT;
 TITLE3 'LEVENE"S TEST';

/* Perform LSD on rankits. */

PROC GLM DATA=Q.CSS;
 CLASS TRT;
 MODEL RANKIT=TRT;
 MEANS TRT/LSD ALPHA=.l;
 TITLES 'LSD TEST';

/* Perform t-tests for each dredged sediment-reference sediment comparison */
/* using rankits. */

PROC TTEST DATA=T1;
 CLASS TRT; VAR RANKIT;
 TITLE3 'T-TEST';
PROC TTEST DATA=T2;
 CLASS TRT; VAR RANKIT;
PROC TTEST DATA=T3;
 CLASS TRT; VAR RANKIT;

/* Calculate power of an LSD test to detect true population differences */
/* 10, 25, 50, and 100% above the reference mean Css. */

DATA Cl;
 SET Wl;
 IF _TYPE_A='ERROR' THEN DELETE;
 MSE=SS/DF;
 MERGEVAR=1;
 KEEP MSB DF MERGEVAR;
DATA C2;
 SET Y;
 IF TRTA=1 THEN DELETE;
DATA C3;
 MERGE Cl C2;
 TALPHA=TINV(.95,DF);
 LABEL    N='NO. OF REPLICATES, N'
      MEANCSS='REFERENCE MEAN Css'
      MSE='MEAN SQUARE ERROR, MSB'
      DF='DEGREES OF FREEDOM, DF'
      TALPHA='T VALUE FOR  (1-ALPHA=0.95,DF)';
 TITLE2 'POWER OF LSD TO DETECT A TRUE POPULATION DIFFERENCE (D)';
 TITLES 'ABOVE REFERENCE MEAN Css';
PROC PRINT LABEL NOOBS; VAR N MEANCSS MSE DF TALPHA;
DATA C4;
 SET C3;
                                                                   DRAFT

-------
                                        D-85

 DO PCTDIFF=10,25,50,100,200,300;
  SEDCSS=MEANCSS+((PCTDIFF/100)*MEANCSS);
  D=SEDCSS-MEANCSS;
  TBETA=D*SQRT(N/(2*MSE))-TALPHA;
  POWER=PROBT(TBETA,DF);
  OUTPUT;
  END;
 LABEL    PCTDIFF='% INCREASE IN Css ABOVE REFERENCE'
      SEDCSS='DREDGED SEDIMENT Css'
      TBETA='T VALUE FOR (1-BETA,DF)'
      POWER='POWER (1-BETA)';
PROC PRINT LABEL NOOBS; VAR PCTDIFF SEDCSS D TBETA POWER;
 TITLE 'POWER OF LSD TO DETECT % INCREASE IN Css ABOVE REFERENCE';
 TITLE2 'MEAN Css GIVEN N, MSB AND DF SHOWN ABOVE';
DATA C5;
 SET C3;
 DO POWER=.5,.6,.7,.8,.9,.95,.99;
  TBETA=TINV(POWER,DF);
  D=((TBETA+TALPHA)*SQRT(2*MSE))/SQRT(N);
  SEDCSS=MEANCSS+D;
  PCTDIFF=(D*100)/MEANCSS ;
  OUTPUT;
  END;
 LABEL    SEDCSS='DREDGED SEDIMENT Css'
      PCTDIFF='% INCREASE IN Css ABOVE REFERENCE'
      TBETA='T VALUE FOR (1-BETA,DF)'
      POWER='POWER (1-BETA)';
PROC PRINT LABEL NOOBS; VAR POWER D SEDCSS PCTDIFF TBETA;
 TITLE 'MINIMUM DREDGED SEDIMENT Css THAT CAN BE DETECTED BY LSD';
 TITLE2 'AS SIGNIFICANT GIVEN SPECIFIED POWER AND N, MSB, AND DF SHOWN ABOVE';

/* Calculation of upper confidence limits (UCL) for comparison of mean */
/* dredged sediment Css with an action level. */

DATA D;
 MERGE Cl Y; BY MERGEVAR;
 IF TRT=1 THEN DELETE;
 TALPHA1=TINV(.95,DF);
 TALPHA2=TINV(.95,N-l);
 UCL1=MEANCSS+TALPHA1*(SQRT(MSE/N));
 UCL2=MEANCSS+TALPHA2 *(SQRT(S2/N));
 DMIN1=TALPHA1*SQRT(MSE/N);
 DMIN2=TALPHA2*SQRT(S2/N);
 LABEL    UCL1='UCL (EQUAL VARIANCES)'
      UCL2='UCL (UNEQUAL VARIANCES)'
      TALPHA1='T VALUE FOR (1-ALPHA=.95,DF)'
      TALPHA2='T VALUE FOR (1-ALPHA=.95,N-1)'
      DMIN1='MINIMUM SIGNIFICANT DIFFERENCE'
      DMIN2='MINIMUM SIGNIFICANT DIFFERENCE'
      MSE='MEAN SQUARE ERROR'
      S2='VARIANCE'
      MEANCSS='MEAN DREDGED SEDIMENT Css';
 TITLE 'COMPARISON OF MEAN DREDGED SEDIMENT Css WITH ACTION LEVEL:';
PROC PRINT LABEL NOOBS; VAR TRT MEANCSS UCL1 MSB TALPHA1 DF DMIN1;
 TITLE2 'UPPER CONFIDENCE LIMITS (UCL) WHEN VARIANCES ARE EQUAL';
PROC PRINT LABEL NOOBS; VAR TRT MEANCSS UCL2 S2 TALPHA2 N DMIN2;
 TITLE2 'UPPER CONFIDENCE LIMITS (UCL) WHEN VARIANCES ARE UNEQUAL';

/* Calculate power of dredged sediment-action level comparisons using */
/* MSB given 10, 20, 30, 40, and 50% decreases in mean Css below */
/* action level. */

DATA Dl;
                                                                   DRAFT

-------
                                        D-86

 SET C3;
 ACTION=2;
 DO PCTDIFF=10 TO 50 BY 10;
  D=PCTDIFF*ACTION/100;
  SEDCSS=ACTION-D;
  TBETA=D*SQRT(N/MSE)-TALPHA;
  POWER=PROBT(TBETA,DF);
  OUTPUT;
  END;
 LABEL    PCTDIFF='% DECREASE BELOW ACTION LEVEL'
      SEDCSS='DREDGED SEDIMENT CBB'
      TBETA='T VALUE FOR (1-BETA,DF)'
      POWER='POWER (1-BETA)';
PROC PRINT NOOBS LABEL; VAR PCTDIFF SEDCSS D TBETA POWER;
 TITLE 'POWER TO DETECT % DECREASE IN Css BELOW';
 TITLE2 'ACTION LEVEL OF 2 ug/g GIVEN N, MSB AND DF SHOWN ABOVE';


D4.4.3       BIOACCSS.SAS Program Output

                           TIME-SEQUENCED BI©ACCUMULATION
   	 TREATMENT GROUP=REFERENCE 	
                                             CONG.      CONC.
                                              IN         IN
                 OBS    REPLICATE    DAY    TISSUE    SEDIMENT

                   11          2     0.054      0.45
                   21          4     0.441      0.45
                   31          7     0.687      0.45
                   4        1         10     0.037      0.45
                   5        1         18     0.856      0.45
                   6        1         28     0.514      0.45
                   72          2     0.163      0.45
                   82          4     0.797      0.45
                   92          7     0.177      0.45
                  10        2         10     0.549      0.45
                  11        2         18     0.598      0.45
                  12        2         28     0.839      0.45
                  13        3          2     0.391      0.45
                  14        3          4     0.203      0.45
                  15        3          7     0.862      0.45
                  16        3         10     0.884      0.45
                  17        3         18     0.016      0.45
                  18        3         28     0.793      0.45
                  19        4          2     0.234      0.45
                  20        4          4     0.564      0.45
                  21        4          7     0.413      0.45
                  22        4         10     0.787      0.45
                  23        4         18     0.806      0.45
                  24        4         28     0.899      0.45
                  25        5          2     0.034      0.45
                  26        5          4     0.018      0.45
                  27        5          7     0.029      0.45
                  28        5         10     0.294      0.45
                  29        5         18     0.119      0.45
                  30        5         28     0.226      0.45
                                                                   DRAFT

-------
D-87
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
79
80
81
1
1
1
1
1
1
2
2
2
2
2
2
3
3
3
3
3
3
4
4
4
4
4
4
5
5
5
5
5
5
1
1
1
1
1
1
2
2
2
2
2
2
3
3
3
3
3
3
4
4
4
J. ViKI
2
4
7
10
18
28
2
4
7
10
18
28
2
4
7
10
18
28
2
4
7
10
18
28
2
4
7
10
18
28
^"DOTI
vjKUU
2
4
7
10
18
28
2
4
7
10
18
28
2
4
7
10
18
28
2
4
7
0.159
0.516
0.881
0.278
0.904
0.172
0.292
0.158
0.317
0.485
1.300
1.049
0.428
0.743
0.270
0.051
0.671
0.476
0.558
0.324
0.562
0.909
0.934
0.712
0.256
0.126
0.603
0.718
1.173
1.245
0.869
0.838
1.246
1.767
1.631
1.178
0.726
0.633
0.816
1.272
1.877
1.721
0.394
0.452
0.897
1.003
1.487
1.366
1.232
0.728
1.639
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
33
33
33
33
33
33
33
33
33
33
33
33
33
33
33
33
33
33
33
33
33
                        DRAFT

-------
             D-88
TIME-SEQUENCED BIOACCUMULATION
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
101
102
103
104
105
106
107
108
109
110
111
112
113
114
115
116
117
118
119
4
4
4
5
5
5
5
5
5
1
1
1
1
1
1
2
2
2
2
2
2
3
3
3
3
3
3
4
4
4
4
4
4
5
5
5
5
5
10
18
28
2
4
7
10
18
28
1.158
1.216
1.513
0.977
1.314
0.688
1.415
1.280
1.843
33
33
33
33
33
33
33
33
33
 TREATMENT GROUP=SEDIMENT 3
2
4
7
10
18
28
2
4
7
10
18
28
2
4
7
10
18
28
2
4
7
10
18
28
2
4
7
10
18
0.745
1.316
1.583
1.578
2.822
1.295
1.703
0.930
2.715
2.268
2.607
2.964
2.045
2.141
1.016
1.756
3.414
2.109
1.855
1.150
2.221
2.899
1.319
2.820
1.135
1.621
2.134
0.890
1.866
44
44
44
44
44
44
44
44
44
44
44
44
44
44
44
44
44
44
44
44
44
44
44
44
44
44
44
44
44
                                       DRAFT

-------
  3

8
N
                                D-89
          TIME-SEQUENCED BIOACCUMULATION
                      TREATMENT GROUP-REFERENCE
                       A"               S                   i
                 D

                       E
                 A


                 "     F
                 F     D               I
                       10                  2O                   3O


                                DAY


         REPLICATE   DOOi   E£ E E 2   L_l_l_3   AAA4   FFF5
Figure D-8.   Plot of Time-Sequenced Bioaccumulation Reference Sediment Example Data by Replicate.
                                                     DRAFT

-------
                               D-90
  3 -
N
C
N
  a-
          TIME-SEQUENCED BIOACCUMULATION
       ~   D
       l_
       g   A


       0   F
                      TREATMENT GROUP-SEDIMENT 1
                       A


                       R
                                      R



                                      6
        REPLICATE
                       10
           20


 DAY

2  l_l_l_3  AAA4
                                                              30
Figure D-9.   Plot of Time-Sequenced Bioaccumulation Dredged Sediment 1 Example Data by Replicate.
                                                    DRAFT

-------
                               D-91
           TIME-SEQUENCED BIOACCUMULATION
                      TREATMENT GROUP-SEDIMENT 2
  3-
 N
 C
 I


 T
            O

            A
        l_
                  A
                        I

                       1O
                                       A
                                DAY
         REPLICATE   D D D
                                          20
                                           AAA
Figure D-10.  Plot of Time-Sequenced Bioaccumulation Dredged Sediment 2 Example Data by Replicate.
                                                   DRAFT

-------
                               D-92
 3 -
N





k1
         TIME-SEQUENCED BIOACCUMULATION
          D

      R   A
                    TREATMENT GROUP-SEDIMENT 3
                      A
                A
                R
                D     D
                     1O
       REPLICATE
                                     D


                                     E
                              DAY
                                        2O
                                   L-I_3   AAA4
                                                            3O
Figure D-ll.  Plot of Time-Sequenced Bioaccumulation Dredged Sediment 3 Example Data by Replicate.
                                                   DRAFT

-------
                                         D-93
(Note:  the following PROCNL1N output is given as an example only for the reference sediment replicate 1. NLIN
      output for the other replicates and sediments has been deleted.)

                            TIME-SEQUENCED  BIOACCUMULATION

    	 TREATMENT GROUP=REFERENCE REPLICATE=1  	
      Non-Linear Least Squares  Grid Search
                                Kl
                          0.300000
                          0.400000
                          0.500000
                          0.200000
                          0.400000
                          0.600000
                          0.300000
                          0.500000
                          0.600000
                          0.700000
                              K2
                        0.210000
                        0.310000
                        0.410000
                        0.110000
                        0.410000
                        0.510000
                        0.310000
                        0.510000
                        0.610000
                        0.610000
                               Dependent  Variable CONC
                        Sum of  Squares
                              0.416199
                              0.425788
                              0.441222
                              0.448040
                              0.454330
                              0.457317
                              0.457654
                              0.460598
                              0.470393
                              0.472661
   Non-Linear Least Squares  DUD  Initialization     Dependent Variable CONC
              DUD              Kl              K2 Sum of Squares
                         0.300000        0.210000       0.416199
                         0.330000        0.210000       0.461659
                         0.300000        0.231000
-3
-2
-1
                               0.405093
Non-Linear Least Squares  Iterative Phase  Dependent Variable CONC Method: DUD
              Iter
                 0
                 1
                 2
                 3
                 4
                 5
                 6
                 7
                 8
               Kl
         0.300000
         0.239451
         0.241348
         0.241312
           237752
         0.237547
         0.237563
         0.237360
         0.237337
0
K2
0.231000
0.178897
0.179839
0.179738
0.176113
0.175943
0.175943
0.175718
0.175695
Sum of Squares
0.405093
0.400026
0.400014
0.400013
0.399983
0.399983
0.399983
0.399983
0.399983
NOTE: Convergence criterion  met.

    Non-Linear Least Squares Summary Statistics
        Source

        Regression
        Residual
        Uncorrected Total

        (Corrected Total)
              DF Sum of Squares

               2   1.2676841229
               4   0.3999828771
               6   1.6676670000
                           Dependent Variable CONC

                           Mean Square

                          0.6338420614
                          0.0999957193
                   0.5505135000
        Parameter
      Estimate
         Asymptotic
         Std. Error
              Kl  0.2373370301  0.22487054331
              K2  0.1756952550  0.21727444929
            Asymptotic 95 %
        Confidence Interval
        Lower         Upper
-.38699524147 0.86166930175
-.42754716392 0.77893767392
                                                                   DRAFT

-------
                                        D-94
   OBS
TREATMENT
  GROUP
                           TIME-SEQUENCED BIOACCUMULATION
                              UPTAKE  DEPURATION  STEADY
                               RATE
                                  RATE
                                STATE
REPLICATE
                            CONSTANT,   CONSTANT,   CONG.,
  kl
k2
Csa
       NORMALIZED
Log10   RANK FOR
 Cea       CBB
     1 REFERENCE
     2 REFERENCE
     3 REFERENCE
     4 REFERENCE
     5 REFERENCE
     6 SEDIMENT
     7 SEDIMENT
     8 SEDIMENT
     9 SEDIMENT
    10 SEDIMENT
    11 SEDIMENT 2
    12 SEDIMENT 2
    13 SEDIMENT 2
    14 SEDIMENT 2
    15 SEDIMENT 2
    16 SEDIMENT 3
    17 SEDIMENT 3
    18 SEDIMENT 3
    19 SEDIMENT 3
    20 SEDIMENT 3
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
23734
30596
53975
31799
04515
05916
01924
24301
05059
02419
01439
00653
00548
03430
02323
01117
01490
09375
02351
00838
0.
0.
0.
0.
0.
0.
0.
2.
0.
0.
0.
0.
0.
0.
0.
0.
0.
1.
0.
0.
17570
20060
40677
16208
08670
42709
04682
20563
24290
06046
31909
11306
11964
87782
56773
25025
23622
97656
45781
13921
0.
0.
0.
0.
0.
0.
1.
0.
0.
1.
1.
1.
1.
1.
1.
1.
2.
2.
2.
2.
60788
68636
59712
88285
23434
55411
64392
44071
83305
60020
48791
90667
51129
28959
35040
96371
77595
08697
25943
64810
-0.
-0.
-0.
-0.
-0.
-0.
0.
-0.
-0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
21618
16345
22394
05411
63015
25641
21588
35584
07933
20418
17258
28028
17935
11045
13046
29308
44341
31952
35400
42293
-0
-0
-0
-0
-1
-1
0
-1
-0
0
0
0
0
-0
-0
0
1
0
1
1
.74414
.58946
.91914
.31457
.86824
.12814
.44777
.40341
.44777
.31457
.06193
.58946
.18676
.18676
.06193
.74414
.86824
.91914
.12814
.40341
      TREATMENT
 OBS    GROUP
           N
                           TIME-SEQUENCED BIOACCUMULATION
     MEAN
     CBB
  1   REFERENCE   5  0.60171
  2   SEDIMENT 1  5  1.01440
  3   SEDIMENT 2  5  1.50917
  4   SEDIMENT 3  5  2.34683
          STANDARD
VARIANCE    ERROR
                        0.05531
                        0.32833
                        0.05797
                        0.12421
                       0.10517
                       0.25625
                       0.10768
                       0.15761
          MEAN
          LoglO
           CBB
         VARIANCE
          OF LOGS
                    -0.25757  0.047978
                    -0.05430  0.068052
                     0.17462  0.004314
                     0.36659  0.004214
           STANDARD
           ERROR OF
             LOGS

            0.09796
            0.11666
            0.02937
            0.02903
Variable=RESID
Variable=RESIDLOG
                    TIME-SEQUENCED BIOACCUMULATION
                   SHAPIRO-WILKS TEST FOR NORMALITY

                         UNIVARIATE PROCEDURE

            N                20
            W:Normal   0.963283  Prob
-------
                                        D-95
                           TIME-SEQUENCED BIOACCUMULATION
                                    LEVENE'S  TEST
                           General  Linear  Models Procedure
Dependent Variable:

Source

Model

Error

Corrected Total

Dependent Variable:

Source

Model

Error

Corrected Total
ABSDEV
DF
3
16
19
ABSLOG
DF
3
16
19
ABSOLUTE DEVIATIONS
Sum of
Squares
0.37008913
0.41648071
0.78656984
ABSOLUTE DEVIATIONS
Sum of
Squares
0.09646576
0.13965602
0.23612178
FROM CSS MEAN
Mean
Square F
0.12336304
0.02603004

Value
4.74


FROM logCss MEAN
Mean
Square F Value
0.03215525
0.00872850

3.68


                              Pr > F

                              0.0150
                              Pr > F

                              0.0344
                           TIME-SEQUENCED BIOACCUMULATION
                            LSD TEST  (UNTRANSFORMED  DATA)
                           General  Linear  Models  Procedure

                           T tests  (LSD)  for  variable:  CSS

            NOTE: This test  controls the type I  comparisonwise error  rate  not the
                  experimentwise error rate.

                          Alpha= 0.1   df=  16  MSE= 0.141456
                              Critical Value  of T= 1.75
                        Least Significant Difference= 0.4153

             Means with the same letter are not significantly  different.
                   T Grouping

                            A

                            B

                            C
                            C
                            C
 Mean      N  TRT

2.347      5  SEDIMENT 3

1.509      5  SEDIMENT 2

1.014      5  SEDIMENT 1

0.602      5  REFERENCE
                                                                  DRAFT

-------
                                        D-96
                           TIME-SEQUENCED BIOACCUMULATION
                           LSD  TEST  (LOG-TRANSFORMED DATA)
                           General Linear Models Procedure

                         T tests (LSD) for variable:  LOGCSS

            NOTE: This test  controls the type I  comparisonwise error rate  not the
                  experimentwise error rate.

                          Alpha= 0.1  df= 16  MSE= 0.03114
                             Critical Value of T=  1.75
                        Least Significant Difference= 0.1949

            Means with the same  letter  are  not significantly  different.
                   T Grouping

                            A
                            A
                            A

                            B

                            C
                    Mean      N  TRT

                   0.367      5  SEDIMENT 3

                   0.175      5  SEDIMENT 2

                  -0.054      5  SEDIMENT 1

                  -0.258      5  REFERENCE
Variable: CSS

       TRT       N
    TIME-SEQUENCED BIOACCUMULATION
                T-TEST

           TTEST  PROCEDURE

STEADY STATE CONC., Css

           Mean             Std Dev
                  Std Error
REFERENCE
SEDIMENT 1
Variances
Unequal
Equal
5
5
T
-1.4899
-1.4899
DF
5.3
8.0
0.60171086 0.23517166
1.01440008 0.57300347
Prob> j T j
0.1935
0.1746
0.10517196
0.25625494

For HO: Variances are equal, F' =5.94    DF = (4,4)
                                 Prob>F'
               0.1127
Variable: LOGCSS

       TRT       N
LoglO Css
           Mean
Std Dev
Std Error
REFERENCE
SEDIMENT 1
Variances
Unequal
Equal
5
5
T
-1.3343
-1.3343
-0
-0
DF
7.8
8.0
.25756572 0.21903881
.05430384 0.26086789
Prob> | T j
0.2200
0.2188
0.09795713
0.11666367

For HO: Variances are equal, F' = 1.42    DF = (4,4)
                                 Prob>F'
               0.7431
                                                                   DRAFT

-------
                                        D-97
Variable: CSS

       TRT       N
           TTEST PROCEDURE
STEADY STATE CONC.,  CBB

           Mean
Std Dev
Std Error
REFERENCE 5
SEDIMENT 2 5
Variances T
Unequal -6.0289
Equal -6.0289
For HO: Variances are
Variable: LOGCSS
TRT N
REFERENCE 5
SEDIMENT 2 5
Variances T
Unequal -4.2261
Equal -4.2261
For HO: Variances are

Variable: CSS
TRT N
REFERENCE 5
SEDIMENT 3 5
Variances T
Unequal -9.2100
Equal -9.2100
For HO: Variances are
Variable: LOGCSS
TRT N
REFERENCE 5
SEDIMENT 3 5
Variances T
Unequal -6.1091
Equal -6.1091
For HO: Variances are
0.60171086 0.23517166
1.50916957 0.24077410
DF Prob> j T |
8.0 0.0003
8.0 0.0003
equal, F' - 1.05 DF = (4,4) Prob>F' =
LoglO Css
Mean Std Dev
-0.25756572 0.21903881
0.17462207 0.06568351
DF Prob> j T j
4.7 0.0097
8.0 0.0029
equal, F' = 11.12 DF = (4,4) Prob>F'
TTEST PROCEDURE
STEADY STATE CONC., Css
Mean Std Dev
0.60171086 0.23517166
2.34683295 0.35243662
DF Prob>|T|
7.0 0.0001
8.0 0.0000
equal, F' = 2.25 DF = (4,4) Prob>F ' =
LoglO Css
Mean Std Dev
-0.25756572 0.21903881
0.36658794 0.06491256
DF Prob> | T !
4.7 0.0023
8.0 0.0003
equal, F' = 11.39 DF = (4,4) Prob>F'
0.10517196
0.10767745


0.9647

Std Error
0.09795713
0.02937456


= 0.0386


Std Error
0.10517196
0.15761445


0.4525

Std Error
0.09795713
0.02902978


= 0.0370
                                                                  DRAFT

-------
                                        D-98

                           TIME-SEQUENCED BIOACCUMULATION
                              Css CONVERTED TO RANKITS
                          SHAPIRO-WILKS TEST FOR NORMALITY

                                UNIVARIATE PROCEDURE
Variable=RESID
                   N                20
                   WrNormal   0.970187  Prob F

Model                    3       0.52458729      0.17486243     1.88    0.1741

Error                   16       1.49037397      0.09314837

Corrected Total         19       2.01496126


                           TIME-SEQUENCED BIOACCUMULATION
                              CSS CONVERTED TO RANKITS
                                      LSD TEST

                           General  Linear  Models Procedure
                         T tests (LSD) for variable: RANKIT

            NOTE: This  test  controls the type  I  comparisonwise error  rate  not the
                  experimentwise error rate.

                       Alpha= 0.1  df= 16  MSE= 0.33088
                           Critical Value of T= 1.75
                     Least Significant Difference= 0.6352

          Means with the same letter are not significantly different.

                   T Grouping              Mean      N  TRT

                            A             1.213      5  SEDIMENT 3

                            B             0.118      5  SEDIMENT 2
                            B
                    C       B            -0.443      5  SEDIMENT 1
                    C
                    C                    -0.887      5  REFERENCE
                                                                   DRAFT

-------
                                        D-99
Variable: RANKIT

       TRT       N
                TIME-SEQUENCED BIOACCUMULATION
                   Css CONVERTED TO RANKITS
                            T-TEST

                        TTEST  PROCEDURE

            RANK FOR VARIABLE CSS

                       Mean             Std Dev
                                             Std Error
 REFERENCE
SEDIMENT 1

Variances
              DF
   -0.88710960
   -0.44339680

       Prob> j T j
                                     0.59170982
                                     0.83054481
Unequal
Equal
-0.9729
-0.9729
7.2
8.0
0.3621
0.3591
For HO: Variances are equal, F' = 1.97
                               DF
                       (4,4)
                       Prob>F'
                                   0.26462068
                                   0.37143093
0.5275
Variable: RANKIT

       TRT       N
                        TTEST  PROCEDURE

            RANK FOR VARIABLE CSS

                       Mean             Std Dev
                                             Std Error
REFERENCE
SEDIMENT 2
Variances
Unequal
Equal
5
5
T
-3.3918
-3.3918
-0.
0.
DF
5.9
8.0
88710960 0.59170982
11789116 0.29807434
Prob> ! T j
0.0151
0.0095
0.26462068
0.13330290

For HO: Variances are equal, F' = 3.94
                               DF
                       (4,4)
                       Prob>F'
0.2126
Variable: RANKIT

       TRT       N
                        TTEST  PROCEDURE

            RANK FOR VARIABLE CSS

                       Mean             Std Dev
                                             Std Error
 REFERENCE
SEDIMENT 3

Variances
      5         -0.88710960
      5          1.21261524

      T       DF    Prob>jT|
Unequal
Equal
-6.3607
-6.3607
7.4
8.0
0.0003
0.0002
                        0.59170982
                        0.44129976
For HO: Variances are equal, F' =1.80    DF = (4,4)
                                             Prob>F'
                                   0.26462068
                                   0.19735525
                                          0.5839
                                                                  DRAFT

-------
                                      D-100
                          TIME-SEQUENCED BIOACCUMULATION
             POWER OF LSD TO DETECT A TRUE POPULATION DIFFERENCE  (D)
                             ABOVE REFERENCE  MEAN Css
       NO.  OF
    REPLICATES,
         N
       REFERENCE
        MEAN Css

        0.60171
           MEAN
          SQUARE
          ERROR,
           MSE

         0.14146
          DEGREES
            OF
         FREEDOM,
            DF

            16
             T VALUE FOR
          (1-ALPHA=0.95,DF)

               1.74588
             POWER OF LSD TO DETECT % INCREASE IN Css ABOVE REFERENCE
                     MEAN Css GIVEN N, MSE AND DF SHOWN ABOVE
        % INCREASE
           IN Css
           ABOVE
         REFERENCE

             10
             25
             50
            100
            200
            300
           DREDGED
          SEDIMENT
             Css

           0.66188
           0.75214
           0.90257
            .20342
            ,80513
1.
1.
           2.40684
0.06017
0.15043
0.30086
0.60171
1.20342
1.80513
  T VALUE
    FOR
(1-BETA,DF)

  -1.49293
  -1.11349
  -0.48110
   0.78369
   3.31327
   5.84285
  POWER
(1-BETA)

 0.07746
 0.14097
 0.31848
 0.77767
 0.99780
 0.99999
             MINIMUM DREDGED SEDIMENT Css THAT CAN BE DETECTED BY LSD
       AS SIGNIFICANT GIVEN SPECIFIED POWER AND N, MSE, AND DF  SHOWN ABOVE
           POWER
         (1-BETA)

           0.50
           0.60
           0.70
           0.80
           0.90
           0.95
           0.99
         0.41529
         0.47657
         0.54256
         0.62097
         0.73327
         0.83059
         1.02983
                     DREDGED
                    SEDIMENT
                       Css
           .01700
           .07828
           ,14427
           .22268
           .33498
          1.43230
          1.63154
          % INCREASE
            IN Css
            ABOVE
          REFERENCE

            69.019
            79.202
            90.169
           103.201
           121.864
           138.038
           171.150
              T VALUE
                FOR
            (1-BETA,DF)

              0.00000
              0.25760
              0.53501
              0.86467
              1.33676
              1.74588
              2.58349
TREATMENT
  GROUP

SEDIMENT 1
SEDIMENT 2
SEDIMENT 3
            COMPARISON OF MEAN DREDGED SEDIMENT Css WITH ACTION LEVEL:
              UPPER CONFIDENCE LIMITS (UCL) WHEN VARIANCES ARE EQUAL
  MEAN
 DREDGED
SEDIMENT  UCL (EQUAL
   Css    VARIANCES)
 1.01440
 1.50917
 2.34683
 1.30806
 1.80283
 2.64049
             MEAN
            SQUARE     T VALUE FOR
            ERROR   (1-ALPHA=.95,DF)
0.14146
0.14146
0.14146
   1.74588
   1.74588
   1.74588
       MINIMUM
     SIGNIFICANT
 DF   DIFFERENCE

 16    0.29366
 16    0.29366
 16    0.29366
                                                                  DRAFT

-------
                                      D-101
            COMPARISON OF MEAN DREDGED SEDIMENT CBS WITH ACTION LEVEL:
             UPPER CONFIDENCE LIMITS (UCL) WHEN VARIANCES ARE UNEQUAL
TREATMENT
GROUP
MEAN
DREDGED
SEDIMENT
CBB
UCL
(UNEQUAL
VARIANCES )
SEDIMENT 1
SEDIMENT 2
SEDIMENT 3
1.01440
1.50917
2.34683
1.56070
1.73872
2.68284
             T VALUE FOR
VARIANCE  (1-ALPHA=.95,N-1)  N

 0.32833       2.13185       5
 0.05797       2.13185       5
 0.12421       2.13185       5
           MINIMUM
         SIGNIFICANT
          DIFFERENCE

           0.54630
           0.22955
           0.33601
                      POWER TO  DETECT  % DECREASE  IN Css BELOW
              ACTION LEVEL OF 2 ug/g GIVEN N, MSB AND DF SHOWN ABOVE
          % DECREASE    DREDGED
            BELOW      SEDIMENT
         ACTION LEVEL     Css
              10
              20
              30
              40
              50
             1.8
             1.6
             1.4
             1.2
             1.0
           0.2
           0.4
           0.6
           0.8
           1.0
          T VALUE
            FOR
        (1-BETA,DF)

          -0.55682
           0.63224
           1.82131
           3.01037
           4.19943
  POWER
(1-BETA)

 0.29268
 0.73192
 0.95634
 0.99585
 0.99966
                                                                 DRAFT

-------
	D-102	

D5.0       REFERENCES


Alldredge, J.R.  1987. Sample size for monitoring of toxic chemical sites. Environ. Monit. Assess. 9:143-154.

Clarke, J.U. and V.A. McFarland. 1991.  Assessing bioaccumulation in aquatic organisms exposed to contam-
        inated sediments. Miscellaneous Paper D-91-2, U.S. Army Engineer Waterways Experiment Station,
        Vicksburg, MS.  77pp.

Conover, W.J.  1980. Practical Nonparametric Statistics.  2nd Ed. John Wiley & Sons, New York, NY.  493
        pp.

Conover, W.J.,  M.E. Johnson and M.M. Johnson.  1981.  A comparative study of tests for homogeneity of
        variances, with applications to the outer continental shelf bidding data. Technometrics 23:351-361.

Dixon, WJ. and F.J. Massey, Jr. 1983.  Introduction to Statistical Analysis.  4th Ed. McGraw-Hill Book
        Company, New York, NY.  678  pp.

Draper, N.R. and H. Smith.  1981.  Applied Regression Analysis. 2nd Ed. John Wiley and Sons, Inc., New
        York, NY.  709pp.

Dunnett, C.W.  1955.  Multiple comparison procedure for comparing several  treatments  with a control. J.
        Amer. Statist. Assoc. 50:1096-1121.

Gelber, R.D., P.T. Lavin, CR. Mehta and D.A.  Schoenfeld. 1985. Chapter 5: Statistical analysis. In G.M.
        Rand and S.R. Petrocelli (Eds.), Fundamentals of Aquatic Toxicology: Methods and Applications.
        Hemisphere Publishing Corp., Washington, DC,  pp. 110-123.

Gill, J.L. 1978. Design and Analysis of Experiments in the Animal and Medical Sciences. Vol. 3. Appendi-
        ces. The Iowa State University Press, Ames, LA.  173 pp.

Hamilton,  M.A., R.C. Russo and R.V. Thurston.  1977. Trimmed Spearman-Karber method for estimating
        median lethal concentrations in toxicity bioassays. Environ. Sci. Technol.  11:714-719.

Hurlbert, S.H. 1984. Pseudoreplication and the design of ecological field experiments. Ecol. Monogr. 54:187-
        211.

Kleinbaum, D.G. and L.L. Kupper.  1978. Applied Regression Analysis and Other Multivariable Methods.
        Duxbury Press, North Scituate, MA.  556  pp.

Lilliefors, H.W. 1967. On the Kolmogorov-Smirnov test for normality with  mean and variance unknown.
        J. Amer. Statist. Assoc. 62:399-402.

McClave, J.T. and F.H. Dietrich, II.  1979. Statistics. Dellen Publishing Company, San Francisco, CA.  681
        pp.

McFarland, V.A. and J.U. Clarke. 1987.  Simplified approach for evaluating bioavailability of neutral organic
        chemicals in sediment.  Environmental  Effects of Dredging Technical Note EEDP-01-8, U.S. Army
        Engineer Waterways Experiment Station,  Vicksburg, MS.

Rohlf, F.J. and R.R. Sokal.  1969. Statistical Tables. W.H. Freeman and Company, New York, NY. 253pp.

SAS Institute, Inc. 1988a. SAS» Language Guide for Personal Computers, Release 6.03 Edition. SAS Insti-
        tute, Inc., Cary, NC.  558 pp.
                                                                                   DRAFT

-------
_ D-103 _

SAS Institute, Inc.  1988b. SAS/STAT™ User's Guide, Release 6.03 Edition.  SAS Institute, Inc., Gary, NC.
       1028pp.

SAS Institute, Inc.  1988c.  SAS/GRAPH* User's Guide, Release 6.03 Edition. SAS Institute, Inc., Gary, NC.
       549 pp.

Satterthwaite, F.W.  1946. An approximate distribution of estimates of variance components.  Biom. Bull.
       2:110-114.

SCI. 1989. PCNONLIN Version 3.0.  Statistical Consultants, Inc., 300 E. Main Street, Quality Place, Suite
       400, Lexington, KY  40507-1539.

Shapiro, S.S. and  M.B. Wilk.   1965.  An analysis of variance test for normality (complete samples).
       Biometrika 52:591-611.

Snedecor,  G.W. and G.C.  Cochran.  1989.  Statistical Methods.  8th Ed.  The Iowa State University Press,
       Ames, IA.  507 pp.

Sokal, R.R. and F.J. Rohlf. 1981. Biometry, 2nd Edition. W.H. Freeman and Company, San Francisco, CA.
       859 pp.

Steel, R.G.D. and J.H. Tome. 1980. Principles and Procedures of Statistics, 2nd Edition. McGraw-Hill Book
       Company, New York, NY.  633 pp.

Steinberg, D.  1988.  PROBIT: A Supplementary Module for SYSTAT and SYGRAPH.  SYSTAT Inc.,
       Evanston, IL.  16  pp.

Steinberg, D. and P. Colla. 1991. LOGIT: A Supplementary Module for SYSTAT. SYSTAT Inc., Evanston,
       IL.  225pp.
Stephan, C.E.  1977.  Methods for Calculating an LCjo.  In F.L. Mayer and J.A. Hamelink (Eds.), Aquatic
        Toxicology and Hazard Evaluation. ASTM Spec. Tech. Publ. 634.  American Society for Testing and
        Materials, Philadelphia, PA, pp. 65-84.

USEPA. 1985.  Methods for measuring the acute toxicity of effluents to freshwater and marine organisms.
        3rd Ed.  Environmental Protection Agency, Environmental  Monitoring and Support Laboratory,
        Cincinnati, OH.  EPA-600/4-85/013.

USEPA. 1989.  Short-term methods for estimating the chronic toxicity of effluents and receiving waters to
        aquatic organisms.  2nd Ed.  U.S. Environmental Protection Agency Rept.  No.  EPA/600/4-89/001.
        262 pp.

Wilkinson, L. 1990.  SYSTAT: The System for Statistics. SYSTAT Inc., Evanston, IL. 677 pp.

Winer, B.J.  1971. Statistical Principles in Experimental Design.  2nd Ed. McGraw-Hill Book Company, New
        York, NY.  907pp.

Zar, J.H. 1984.  Biostatistical Analysis. 2nd Ed. Prentice-Hall, Inc.,  Englewood Cliffs, NJ. 717 pp.
                                                                                 DRAFT

-------
D-104
                             DRAFT

-------
       APPENDIX £
    SUMMARY OF TEST
   CONDITIONS AND TEST
ACCEPTABILITY CRITERIA FOR
     TIER III BIOASSAYS
                               DRAFT

-------
  Acute Tox/crty
Water Column Tests
                               DRAFT

-------
                                             E-l
     SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR MYSID
  SHRIMP, Mysidopsis bahia, M. bigelowi, M. almyra, Neomysis americana, Holmesimysis costata, ACUTE
                             TOXICITY WATER COLUMN TESTS
1.   Test type:
2.   Test duration:
3.   Temperature:
4.   Salinity:

5.   Light quality:
6.   Light intensity:
7.   Photoperiod:
8.   Test chamber size:
9.   Test solution volume:
10.  Renewal of test solutions:
11.  Age of test organisms:
12.  No. organisms per test chamber:
13.  No. replicate chambers per concentration:
14.  No. organisms per concentration:
15.  Feeding regime:
16.  Test chamber cleaning:
17.  Test solution aeration:
18.  Dilution water:
19.  Test concentrations:

20.  Dilution series:
21.  Endpoint:
Static Non-renewal
96h
20±1°C: or 25±1°C for
  Mysidopsis bahia
  Mysidopsis bigelowi
  Mysidopsis almyra
20±l°Cfor
  Neomysis americana
12±l°Cfor
  Holmesimysis costata
25-30 %o ±10% except for Holmesimysis costata which
is to be 32-34 %o± 10%
Ambient Laboratory
10-20 uE/m2/s (50-100 ft-c)
16L/8D
250 mL minimum
200 mL minimum
None
1 - 5 d; 24 h range in age
10 minimum
5 minimum
50 minimum
Anemia nauplii are made available while holding
prior to the test; feed 0.2 mL of concentrated
suspension of Anemia nauplii s24 h old, daily
(approximately 100 nauplii per mysid)
None
If needed to maintain DO> 40% for:
  Mysidopsis bahia
  Mysidopsis bigelowi
  Mysidopsis almyra
  Neomysis americana
and DO> 60% saturation for:
  Holmesimysis costata
(< 100 bubbles/min.)
Natural seawater or modified GP2, Forty Fathoms*
or equivalent, artificial seawater prepared with
Millipore MILLI-Q* or equivalent or deionized
water
Three concentrations for site sediment, and control
water
100%, 50%,  10%
Survival

-------
                                             E-2
 22. Sampling and sample holding requirements:   <6 wk (sediment); elutriates are to be used within
                                              24 h of preparation
 23. Sample volume required:

 24. Test acceptability criterion:
1 L per site

* 90% survival in controls
REFERENCE:

USEPA.  1991. Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater
    and Marine Organisms, 4th Ed.  EPA/600/4-90/027.

-------
                                             E-3
      SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR GRASS
              SHRIMP, Palaemonetes sp., ACUTE TOXICITY WATER COLUMN TESTS
 1.  Test type:
 2.  Test duration:
 3.  Temperature:
 4.  Salinity:
 5.  Light quality:
 6.  Light intensity:
 7.  Photoperiod:
 8.  Test chamber size:
 9.  Test solution volume:
 10. Renewal of test solutions:
 11. Age of test organisms:
 12. No. organisms per test chamber:
 13. No. replicate chambers per concentration:
 14. No. organisms per concentration:
 15. Feeding regime:
 16. Test chamber cleaning:
 17. Test solution aeration:

 18. Dilution water:
 19. Test concentrations:

 20. Dilution series:
 21. Endpoint:
 22. Sampling and sample holding requirements:

 23. Sample volume required:
 24. Test acceptability criterion:
Static Non-renewal
96h
25±1°C
30-35 %o ±10%
Ambient Laboratory
10-20 uE/m2/s (50-100 ft-c)
16L/8D
1 L minimum
750 mL minimum
None
1-4 d from hatch
10 minimum
5 minimum
50 minimum
None
None
If needed to maintain DO> 40% saturation
(< 100 bubbles/min.)
Natural seawater or modified GP2, Forty Fathoms*
or equivalent, artificial seawater prepared with
Millipore MILLI-Q* or equivalent or deionized
water
Three concentrations for site sediment, and control
water
100%, 50%, 10%
Survival
<6 wk (sediment); elutriates are to be used within
24 h of preparation
4 L per site minimum
* 90% survival in controls
REFERENCE:
Modified from the mysid acute toxicity water column test published in:
USEPA. 1991. Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater
    and Marine Organisms, 4th Ed. EPA/600/4-90/027.

-------
                                             E-4
  SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR COMMERCIAL
                SHRIMP, Penaeus sp., ACUTE TOXICITY WATER COLUMN TESTS
 1.   Test type:
 2.   Test duration:
 3.   Temperature:
 4.   Salinity:
 5.   Light quality:
 6.   Light intensity:
 7.   Photoperiod:
 8.   Test chamber size:
 9.   Test solution volume:
 10.  Renewal of test solutions:
 11.  Age of test organisms:
 12.  No. organisms per test chamber:
 13.  No. replicate chambers per concentration:
 14.  No. organisms per concentration:
 15.  Feeding regime:
 16.  Test chamber cleaning:
 17.  Test solution aeration:

 18.  Dilution water:
 19. Test concentrations:

 20. Dilution series:
 21. Endpoint:
 22. Sampling and sample holding requirements:

 23. Sample volume required:
 24. Test acceptability criterion:
Static Non-renewal
96 h
25±1°C
30-3596o ±10%
Ambient Laboratory
10-20 uE/m2/s (50-100 ft-c)
16L/8D
SOL
60L
None
8-10 d post larvae
10 minimum
5 minimum
50 minimum
None
None
If needed to maintain DO> 40% saturation
(<  100 bubbles/min.)
Natural seawater or modified GP2, Forty Fathoms*
or equivalent, artificial seawater prepared with
Millipore MILLI-Q* or equivalent or deionized
water
Three concentrations for site sediment, and control
water
100%, 50%, 10%
Survival
<6 wk (sediment); elutriates are to be used within
24 h of preparation
20 L for site sediment
* 90% survival in controls
REFERENCE:
Modified from the mysid shrimp acute toxicity water column test published in:
USEPA. 1991. Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater
    and Marine Organisms, 4th Ed. EPA/600/4-90/027.

-------
                                             E-5
       SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE
     CLADOCERANS, Daphnia magna AND D. pulex, ACUTE TOXICITY WATER COLUMN TESTS
 1.  Test type:
 2.  Test duration:
 3.  Temperature:
 4.  Salinity:
 5.  Light quality:
 6.  Light intensity:
 7.  Photoperiod:
 8.  Test chamber size:
 9.  Test solution volume:
 10. Renewal of test solutions:
 11. Age of test organisms:
 12. No. organisms per test chamber:
 13. No. replicate chambers per concentration:
 14. No. organisms per concentration:
 15. Feeding regime:
 16. Test chamber cleaning:
 17. Test solution aeration:
 18. Dilution water:
Static Non-renewal
96h
20 or 25±1°C
0%o
Ambient Laboratory
10-20 uE/m2^ (50-100 ft-c)
16L/8D
30 mL minimum
25 mL minimum
None
Less than 24 h old
5 minimum
5 minimum
25 minimum
Feed YCT* and Selenastrum while holding prior to
the test; newly-released young should have food
available a minimum of 2 h prior to use in a test;
add 0.2 mL each of YCT and Selenastrum at -2 h and
at48h.
None
None
Moderately hard synthetic water prepared using
Millipore MILLI-Q* or equivalent deionized water
and reagent grade chemicals or 20% DMW, receiving
water, or synthetic water modified to reflect receiving
water hardness
Three concentrations for site sediment, and control
water
100%, 50%, 10%
Survival
<6 wk (sediment); elutriates are to be used within
24 h of preparation
1 L per site
* 90% survival in controls
 19. Test concentrations:

 20. Dilution series:
 21. Endpoint:
 22. Sampling and sample holding requirements:

 23. Sample volume required:
 24. Test acceptability criterion:
*   Slurry of Yeast, Cereal flakes, Trout chow.
REFERENCE:
USEPA 1991. Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater
    and Marine Organisms, 4th Ed. EPA/600/4-90/027.

-------
                                             E-6
      SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE
          CLADOCERAN, Ceriodaphnia dubia, ACUTE TOXKTTY WATER COLUMN TESTS
 1.  Test type:
 2.  Test duration:
 3.  Temperature:
 4.  Salinity:
 5.  Light quality:
 6.  Light intensity:
 7.  Photoperiod:
 8.  Test chamber size:
 9.  Test solution volume:
 10. Renewal of test solutions:
 11. Age of test organisms:
 12. No. organisms per test chamber:
 13. No. replicate chambers per concentration:
 14. No. organisms per concentration:
 15. Feeding regime:
 16. Test chamber cleaning:
 17. Test solution aeration:
 18. Dilution water:
Static Non-renewal
96h
20 or 25±1°C
0%o
Ambient Laboratory
10-20- uE/m2/s (50-100 ft-c)
16L/8D
30 mL minimum
15 mL minimum
None
Less than 24 h old
5 minimum
5 minimum
25 minimum
Feed YCT* and Selenastrum while holding prior to
the test: newly-released young should have food
available a minimum of 2 h prior to use in a test:
add 0.1 mL each of YCT and Selenastrum at -2 h and
at48h
None
None
Moderately hard synthetic water prepared using
Millipore MILLI-Q* or equivalent or deionized
water and reagent grade chemicals, or 20% DMW,
receiving water, or synthetic water modified to reflect
receiving water hardness
Three concentrations for site sediment, and control
water
100%, 50%, 10%
Survival
<6 wk (sediment); elutriates are to be used within
24 h of preparation
1 L per site
2 90% survival in controls
 19.  Test concentrations:

 20.  Dilution series:
 21.  Endpoint:
 22.  Sampling and sample holding requirements:

 23.  Sample volume required:
 24.  Test acceptability criterion:
*  Slurry of Yeast, Cereal flakes, Trout chow.
REFERENCE:
USEPA.  1991. Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater
   and Marine Organisms, 4th Ed. EPA/600/4-90/027.

-------
                                             E-7
   SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR SHEEPSHEAD
  MINNOW, Cyprinodon variegatus, INLAND SILVERSIDE, Menidia beryllina, ATLANTIC SILVERSIDE,
  M. menidia, TIDEWATER SILVERSIDE, M. peninsula*, ACUTE TOXICITY WATER COLUMN TESTS
  1.  Test type:
  2.  Test duration:
  3.  Temperature:
  4.  Salinity:

  5.  Light quality:
  6.  Light intensity:
  7.  Photoperiod:
  8.  Test chamber size:
  9.  Test solution volume:
  10. Renewal of test solutions:
  11. Age of test organisms:

  12. No. organisms per test chamber:
  13. No. replicate chambers per concentration:
  14. No. organisms per concentration:
  15. Feeding regime:

  16. Test chamber cleaning:
  17. Test solution aeration:

  18. Dilution water:
 19. Test concentrations:

 20. Dilution series:
 21. Endpoint:
 22. Sampling and sample holding requirements:

 23. Sample volume required:
 24. Test acceptability criterion:
REFERENCE:
Static Non-renewal
96 h
20 or 25±1°C
Sheepshead minnow: 5-30 %o ± 10%
Silversides: 5-32 %o ± 10%
Ambient Laboratory
10-20 uE/mVs (50-100 ft-c)
16L/8D
250 mL minimum
200 mL minimum
None
Sheepshead minnow: 1 - 14 d; 24-h range in age
Silversides: 9 - 14 d; 24-h range in age
10 minimum
5 minimum
50 minimum
Anemia nauplii are made available while holding
prior to the test; add 0.2 mL Anemia nauplii
concentrate at 48 h
None
If needed to maintain DO> 40% saturation
(< 100 bubbles/min.)
Natural seawater or modified GP2, Forty Fathoms*
or equivalent, artificial seawater prepared with
Millpore MILLI-Q* or equivalent or deionized
water
Three concentrations for site sediment, and control
water
100%, 50%, 10%
Survival
<6 wk (sediment); elutriates are to be used within
24 h of preparation
4 L per site
* 90% survival in controls
USEPA, 1991. Methods for Measuring the Acute Toxiciry of Effluents and Receiving Waters to Freshwater
   and Marine Organisms, 4th Ed.  EPA/600/4-90/027.

-------
                                            E-8
     SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE
   SPECKLED SANDDAB, Citharichthys stigmaeus, ACUTE TOXICITY WATER COLUMN TESTS
1.   Test type:
2.   Test Duration:
3.   Temperature:
4.   Salinity:
5.   Light quality:
6.   Light intensity:
7.   Photoperiod:
8.   Test chamber size:
9.   Test solution volume:
10.  Renewal of test organisms:
11.  Age of test organisms:
12.  No. organisms per test chamber:
13.  No. replicate chambers per concentration:
14  No. organisms per concentration:
15.  Feeding regime:

16.  Test chamber cleaning:
17.  Test solution aeration:

18.  Dilution water:
19. Test concentrations:

20. Dilution series:
21. Endpoint:
22. Sampling and sample holding requirements:

23. Sample volume required:
24. Test acceptability criterion:
Static Non-renewal
96 h
15±2°C
30±2%o
Ambient Laboratory
10-20 /iE/mVs (50-100 ft-c)
16L/8D
30L
20 L
None
Juveniles * 8 cm
10
5 minimum
50 minimum
Anemia nauplii are made available while holding
prior  to the test: add 0.2 mL Anemia nauplii
concentrate at 48 h
None
If needed to maintain DO> 40% saturation
(<  100 bubbles/min.)
Natural seawater or modified GP2, Forty Fathoms*
or equivalent, artificial seawater prepared with
Millipore MILLI-Q* or equivalent or deionized
water
Three concentrations for site sediment, and control
water
100%, 50%, 10%
Survival
<6 wk (sediment); elutriates are to be used within
24 h of preparation
20 L for site sediment
* 90% survival in  controls

-------
                                             E-9

REFERENCE:

Adapted in part from the Menidia sp. protocol published in:

USEPA. 1991. Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater
    and Marine Organisms, 4th Ed. EPA/600/4-90-027.

and from EPA in-house expertise, ERL-Narragansett, RI.

-------
                                                   E-10
1
2
3
4
        SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR
            GRUNION, Leunsthes tenuis, ACUTE TOXICITY WATER COLUMN TESTS
1.   Test type:
2.   Test duration:
3.   Temperature:
4.   Salinity:
5.   Light quality:
6.   Light intensity:
7.   Photoperiod:
8.   Test chamber size:
9.   Test solution volume:
10.  Renewal of test organisms:
11.  Age of test organisms:
12.  No. organisms per test chamber:
13.  No. of replicate chambers per
    concentration:
14.  No. organisms per concentration:
15.  Feeding regime:

16.  Test chamber cleaning:
17.  Test solution aeration:

18.  Dilution water:
        19.  Test concentrations:

        20.  Dilution series:
        21.  Endpoint:
        22.  Sampling and sample holding requirments:

        23.  Sample volume required:
        24.  Test acceptability criterion:
Static Non-renewal
96h
20 or 25±2°C
20-32 %o± 10%
Ambient Laboratory
10-20 ,tE/m2/s (50-100 ft-c)
16L/8D
250 mL  minimum
200 mL  minimum
None
9- 14 d
10
5 minimum

50 minimum
Anemia  nauplii are made available while holding
prior to  the test: add 0.2 mLArtemia nauplii
concentrate at 48 h
None
If needed to maintain DO> 40% saturation
(< 100 bubbles/nun.)
Natural  seawater or modified GP2, Forty Fathoms*
or equivalent, artifical seawater prepared with
Millipore MILLI-Q* or equivalent or deionized
water
Three concentrations for site sediment, and control
water
100%, 50%, 10%
Survival
<6 wk (sediment); elutriates are to be used within
24 h of preparation
20 L for site sediment
i 90% or greater survival in controls
       REFERENCE:
       Adapted in part from the Menidia sp. protocol published in:
       USEPA. 1991. Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater
           and Marine Organisms, 4th Ed.  EPA/600/4-90/027
       and from personal communications with Dr. Doug Middaugh, EPA, ERL-Gulf Breeze, FL.

-------
                                             E-ll
    SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR FATHEAD
      MINNOW, Pimephales promelas, BLUEGILL SUNFISH, Lepomis macrochirus, AND CHANNEL
             CATFISH, Ictalurus punctatus, ACUTE TOXICITY WATER COLUMN TESTS
 1.  Test type:
 2.  Test duration:
 3.  Temperature:
 4.  Salinity:
 5.  Light quality:
 6.  Light intensity:
 7.  Photoperiod:
 8.  Test chamber size:
 9.  Test solution volume:
 10. Renewal of test solutions:
 11. Age of test organisms:

 12. No. organisms per test chamber:
 13. No. replicate chambers per concentration:
 14. No. organisms per concentration:
 15. Feeding regime:

 16. Test chamber cleaning:
 17. Test solution aeration:

 18. Dilution water:
 19. Test concentrations:

 20. Dilution series:
 21. Endpoint:
 22. Sampling and sample holding requirements:

 23. Sample volume required:
 24. Test acceptability criterion:
REFERENCE:
Static Non-renewal
%h
20or25±l°C
0%o
Ambient Laboratory
10-20 uE/m2/s (50-100 ft-c)
16L/8D
250 mL minimum
200 mL minimum
None
Fathead minnow - on order of 4 d; 24 h range in
age. Sunfish and Catfish - on order of 30 d
10 minimum
5 minimum
50 minimum
Anemia nauplii are made available while holding
prior to the test; add 0.2 mL Anemia nauplii
concentrate at 48 h
None
If needed to maintain DO> 40% saturation
(< 100 bubbles/min.)
Moderately hard synthetic water prepared using
Millipore MILLI-Q* or equivalent deionized water
and reagent grade chemicals or 20% DMW, receiving
water, or synthetic water modified to reflect receiving
water hardness
Three concentrations for site sediment, and control
water
100%, 50%, 10%
Survival
<6 wk (sediment); elutriates are to be used within
24 h of preparation
4L per site  minimum
* 90% survival in controls
USEPA.  1991. Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater
    and Marine Organisms, 4th Ed. EPA/600/4-90/027.

-------
                                             E-12
    SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR RAINBOW
             TROUT, Oncorhynchus mykiss, ACUTE TOXICITY WATER COLUMN TESTS
 1.  Test type:
 2.  Test duration:
 3.  Temperature:
 4.  Salinity:
 5.  Light quality:
 6.  Light intensity:
 7.  Photoperiod:
 8.   Test chamber size:

 9.   Test solution volume:
 10.  Renewal of test solutions:
 11.  Age of test organisms:
 12.  No. organisms per test chamber:
 13.  No. replicate chambers per concentration:
 14.  No. organisms per concentration:
 15.  Feeding regime:
 16.  Test chamber cleaning:
 17.  Test solution aeration:

 18.  Dilution water:
 19. Test concentrations:

 20. Dilution series:
 21. Endpoint:
 22. Sampling and sample holding requirements:

 23. Sample volume required:
 24. Test acceptability criterion:
REFERENCE:
Static Non-renewal
96h
12±1°C
0%o
Ambient Laboratory
10-20 uE/m2/s (50-100 ft-c)
16L/8D: Light intensity should be raised gradually
over a 15 min period at the beginning of the
photoperiod, and lowered gradually at the end of the
photoperiod, using a dimmer switch or other suitable
device
5 L minimum, test chambers should be covered to
prevent fish from jumping out
4 L minimum
None
15-30 d (after yolk sac absorption to 30 d)
10 minimum
5 minimum
50 minimum
Feeding not required
None
If needed to maintain DO>  60% saturation
(<  100 bubbles/min.)
Moderately hard synthetic water prepared using
Millipore MILLI-Q* or equivalent deionized water
and reagent grade chemicals or 20% DMW, receiving
water, or synthetic water  modified to  reflect receiving
water hardness
Three concentrations  for site sediment, and control
water
100%, 50%, 10%
Survival
<6 wk (sediment); elutriates are to be used within
24 h of preparation
20 L for site sediment
* 90% survival in controls
USEPA.  1991. Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater
    and Marine Organisms, 4th Ed.  EPA/600/4-90/027.

-------
                                             E-13
     SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR OYSTER,
    Crassostrea virginica, AND MUSSEL, Mytilus edulis, ACUTE TOXICITY WATER COLUMN TESTS
 1.  Test type:
 2.  Test duration:
 3.  Temperature:

 4.  Salinity:
 5.  Light quality:
 6.  Light intensity:
 7.  Photoperiod:
 8.  Test chamber size:*
 9.  Test solution volume:*
 10. Renewal of test solutions:
 11. Age of test organisms:
 12. No. organisms per test chamber:
 13. No. replicate chambers per concentration:
 14. No. organisms per concentration:
 15. Feeding regime:
 16. Test chamber cleaning:
 17. Test solution aeration:
 18. Dilution water:*
Static Non-renewal
48 h
25 ±1° C for Crassostrea virginica
16±1° C for Mytilus edulis
18-32±  1 %o
Ambient Laboratory
10-20 uE/m2/s (50-100 ft-c)
16U8D
1L
500 mL
None
Larvae less than 4 h old
7,500 - 15,000
5 minimum
22,500 - 45,000
None
None
None
Natural seawater or modified GP2, Forty
Fathoms*, artificial seawater prepared with
Millipore MILLI-Q* or equivalent or deionized
water
Three concentrations for site sediment, and
control water
None
Shell development to hinged,  D-shaped
prodissoconch I larva
<6 wk (sediment); elutriates are to be used
within 24 h of preparation
1 L per site
i 70% or greater survival and * 70% shell
development in controls
 19. Test concentrations:

 20. Dilution series:
 21. Endpoint:

 22. Sampling and sample

 23. Sample volume required:
 24. Test acceptability * criterion:
* - Protocol dependent
REFERENCE:
ASTM.  1989. E 724-89.  Standard guide for conducting static acute toxicity tests starting with embryos of
    four species of saltwater bivalve molluscs.  Annual Book of ASTM Standards, Vol. 11.04. American
    Society for Testing and Materials, Philadelphia, PA.

-------
                                            E-14
 SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR SEA URCHINS,
    Strongylocentrotus sp., Lytechinus pictus, AND SAND DOLLAR, Dendraster sp., ACUTE TOXICITY
                                  WATER COLUMN TESTS
 1.  Test type:
 2.  Test duration:
 3.  Temperature:
 4.  Salinity:
 5.  Light quality:
 6.  Light intensity:
 7.  Photoperiod:
 8.  Test chamber size:
 9.  Test solution volume:
 10. Renewal of test solutions:
 11. Age of test organisms:
 12. No. organisms per test chamber:
 13. No. replicate chambers per concentration:
 14. No. organisms per concentration:
 15. Feeding regime:
 16. Test chamber cleaning:
 17. Test solution aeration:
 18. Dilution water:
 19. Test concentrations:

 20. Dilution series:
 21. Endpoint:
 22. Sampling and sample holding requirements:

 23. Sample volume required:
 24. Test acceptability criterion:

REFERENCE:
Static Non-renewal
48 h
12°C
30-32 %o
Ambient Laboratory
10-20 uE/m2/s (50-100 ft-c)
Not essential
20 mL minimum
10 mL minimum
None
s 1 h embryos
2000
3 minimum
6000 minimum
None
None
None
Natural seawater or modified GP2, Forty Fathoms*
or equivalent, artificial seawater prepared using
Millipore MILLI-Q* or equivalent or deionized
water and 3x brine to maintain constant salinity
across tests
Three concentrations for site sediment, and control
water
100%, 50%, 10%
Survival, Embryo Development
<6 wk (sediment); elutriates are to be used within
24 h of preparation
1 L per site
i 70% survival and * 70% normal embryo
development in controls
USEPA.  1990.  Conducting the Sea Urchin Larval Development Test.  ERL-Narragansett Standard
    Operating Procedure 1.03.007.

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                                              E-15
   SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR SEA URCHIN,
    Strongylocentrotus purpuratus, AND SAND DOLLAR, Dendraster excentricus, SPERM CELL ACUTE
                               TOXICITY WATER COLUMN TESTS
 1.  Test type:
 2.  Test duration:

 3.  Temperature:
 4.  Salinity:
 S.  Light quality:
 6.  Light intensity:
 7.  Photoperiod:
 8.  Test chamber size:
 9.  Test solution volume:
 10. Renewal of test solutions:
 11. Age of test organisms:
 12. No. organisms per test chamber:
 13. No. replicate chambers per concentration:
 14. No. organisms per concentration:
 15. Feeding regime:
 16. Test chamber cleaning:
 17. Test solution aeration:
 18. Dilution water:
 19. Test concentrations:

 20. Dilution series:
 21. Endpoint:
 22. Sampling and sample holding requirements:

 23. Sample volume required:
 24. Test acceptability criterion:

REFERENCE:
Static Non-renewal
80 minute (60 minute exposure plus 20 minute
fertilization period)
12°C
30±2%o
Ambient Laboratory
10-20 uE/m2/s (50-100 ft-c)
Not essential
Test tubes 16 x 100 or 125 mm
5mL
None
Fresh eggs and sperm
560,000 sperm/1,120 eggs (100 eggs observed)
3 minimum
300 eggs observed per concentration
None
None
None
Filtered (0.45 /Am): natural seawater or modified
GP2, Forty Fathoms* or equivalent, artificial
seawater prepared using Millipore MILLI-C* or
equivalent or deionized water and 3x brine to
maintain constant salinity  across tests.
Three concentrations for site sediment, and control
water
100%, 50% 10%
Egg fertilization percentage
<6 wk (sediment); elutriates are to be used within
24 h of preparation
1 L per  site
* 50% control fertilization, sperm:egg ratio between
250:1 and 1,000:1
Dinnel, P.A, QJ. Stober, S.C. Crumley and R.E. Nakatani. 1982. Development of a sperm cell toxicity test
    for marine waters. Pp. 82-98 In: Aquatic Toxicity and Hazard Assessment. Fifth Conference. J.G.
    Pearson, R.B. Foster, and W.E. Bishop (Eds.). ASTM STP 766. American Society for Testing and
    Materials, Philadelphia, PA

-------
Acute Toxicity
Sediment Tests
                              DRAFT

-------
                                             E-17
       SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE
                AMPHIPOD, Ampelisca abdita, ACUTE TOXICITY SEDIMENT TESTS
  1.  Test type:
  2.  Test duration:
  3.  Temperature:
  4.  Salinity:
  5.  Light quality:
  6.  Light intensity:
  7.  Photoperiod:
  8.  Test chamber size:
  9.  Test solution volume:
  10. Sediment depth:
  11. Renewal of test solutions:
  12. Age of test organisms:
  13. No. of organisms per test chamber:
  14. No. replicate chambers per sediment:
  15. No. organisms per sediment:
  16. Feeding regime:
  17. Test chamber cleaning:
  18. Test solution aeration:
  19. Dilution water:
 20. Test concentrations:

 21. Dilution series:
 22. Endpoint:
 23. Sampling and sample holding requirements:
 24. Sample volume required:
 25. Test acceptability criterion:
Static Non-renewal*
10 d
20°C
28 to 35 %o
Ambient Laboratory
10-20 uE/m2/s (50-100 ft-c)
Continuous Light
1L
Vol. to 950 mL
4 cm minimum
None*
Immature amphipods, or mature females only
20 to 30
5
100 to 150
None
None
Trickle-flow (< 100 bubbles/min.)
Natural seawater or modified GP2, Forty Fathoms*
or equivalent, artificial seawater prepared using
Millipore MILLI-Q* or equivalent or deionized
water
Site sediment, a reference sediment and a control
sediment
N/A
Survival
<6wk
2L
i 90% survival in controls
REFERENCE:
ASTM.  1991.  E1367-90.  Standard guide for conducting 10-day static sediment toxicity tests with marine
    and estuarine amphipods. Annual Book of ASTM Standards, Vol. 11.04.  American Society for
    Testing and Materials, Philadelphia, PA
*   Static renewal, intermittent flow or continuous flow tests may be used where it is necessary to
    maintain water quality parameters, e.g., dissolved oxygen (DO). For static renewal tests the overlying
    dilution water should be changed every 48 h at a minimum.

-------
                                             E-18
      SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE
            AMPHIPOD, Leptocheirus plumulosus, ACUTE TOXICITY SEDIMENT TESTS
 1.   Test type:
 2.   Test duration:
 3.   Temperature:
 4.   Salinity:
 5.   Light quality:
 6.   Light intensity:
 7.   Photoperiod:
 8.   Test chamber size:
 9.   Test solution volume
 10.  Sediment depth:
 11.  Renewal of test solutions:
 12.  Age of test organisms:
 13.  No. of organisms per test chamber:
 14.  No. replicate chambers per sediment:
 IS.  No. organisms per sediment:
 16.  Feeding regime:
 17.  Test chamber cleaning:
 18.  Test solution aeration:
 19.  Dilution water:
 20. Test concentrations:
 21. Dilution series:
 22. Endpoint:
 23. Sampling and sample holding requirements:
 24. Sample volume required:
 25. Test acceptability criterion:
Static Non-renewal*
10 d
20-25°C
20 %o (range 2 - 32 %o)
Ambient Laboratory
10-20 uE/mVs (50-100 ft-c)
16L/8D
1L
Vol. to 950 mL
2 cm minimum
None*
Mature 3 - 5 mm mixed sexes
20
5
100
None
None
Trickle-flow (< 100 bubbles/min.)
Natural seawater or modified GP2, Forty Fathoms*
or equivalent, artificial seawater prepared with
Millipore MILLI-Q* or equivalent or deionized
water
N/A
N/A
Survival
<6wk
2L
* 90% survival in controls
REFERENCE:
Schlekat, C.E., B.E. McGee and E. Reinharz. 1992. Testing sediment toxicity in Chesapeake Bay using
    the amphipod Leptocheirus plumulosus: an evaluation. Environ. Toxicol. Chem. 11: 225-236.
*   Static renewal, intermittent flow or continuous flow tests may be used where it is necessary to
    maintain water quality parameters, e.g., dissolved oxygen (DO). For static renewal tests the overlying
    dilution water should be changed every 48 h at a minimum.

-------
                                             E-19
      SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE
             AMPHIPOD, Khepoxynius abronius, ACUTE TOXICITY SEDIMENT TESTS
 1.  Test type:
 2.  Test duration:
 3.  Temperature:
 4.  Salinity:
 5.  Light quality:
 6.  Light intensity:
 7.  Photoperiod:
 8.  Test chamber size:
 9.  Test solution volume
 10. Sediment depth:
 11. Renewal of test solutions:
 12. Age of test organisms:
 13. No. of organisms per test chamber:
 14. No. replicate chambers per sediment:
 15. No. organisms per sediment:
 16. Feeding regime:
 17. Test chamber cleaning:
 18. Test solution aeration:
 19. Dilution water:
 20. Test concentrations:
 21. Dilution series:
 22. Endpoint:
 23. Sampling and sample holding requirements:
 24. Sample volume required:
 25. Test acceptability criterion:
Static Non-renewal*
10 d
15 ±3°C
28 %o
Ambient Laboratory
10-20 uE/m2/s (50-100 ft-c)
Continuous Light
1L
Vol. to 950 mL
2 cm minimum
None*
Mature 3 - 5 mm mixed sexes
20
5
100
None
None
Trickle-flow (< 100 bubbles/min.)
Natural  seawater or modified GP2, Forty Fathoms*
or equivalent, artificial seawater prepared with
Millipore MILLI-C* or equivalent or deionized
water
N/A
N/A
Survival
<6 wk
2L
* 90% survival in controls
REFERENCE:
ASTM.  1991.  E1367-90.  Standard guide for conducting 10-day static sediment toxicity tests with marine
    and estuarine amphipods. Annual Book of ASTM Standards, Vol. 11.04. American Society for
    Testing and Materials, Philadelphia, PA.
*   Static renewal, intermittent flow or continuous flow tests may be used where it is necessary to
    maintain water quality parameters, e.g., dissolved oxygen (DO). For static renewal tests the overlying
    dilution water should be changed  every 48 h at a minimum.

-------
                                             E-20
       SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE
             AMPHIPOD, GrandidiereUajaponica, ACUTE TOXICITY SEDIMENT TESTS
 1.   Test type:
 2.   Test duration:
 3.   Temperature:
 4.   Salinity:
 5.   Light quality:
 6.   Light intensity:
 7.   Photoperiod:
 8.   Test chamber size:
 9.   Test solution volume:
 10.  Sediment depth:
 11.  Renewal of test solutions:
 12.  Age of test organisms:

 13.  No. of organisms per test chamber:
 14.  No. replicate chambers per sediment:
 IS.  No. organisms per sediment:
 16.  Feeding regime:

 17.  Test chamber cleaning:
 18.  Test solution aeration:
 19.  Dilution water:
 20. Test concentrations:

 21. Dilution series:
 22. Endpoint:
 23. Sampling and sample holding requirements:
 24. Sample volume required:
 25. Test acceptability criterion:
REFERENCE:
Static Non-renewal*
10 d
15 - 19 ±38C
30 to 35 %o
Ambient Laboratory
10-20 uE/m2/s (50-100 ft-c)
Continuous Light
1L
Vol. to 950 mL
2 cm minimum
None*
Immature amphipods 3-6 mm, no females carrying
embryos
20
5
100
Suspension of finely ground Tetramin and the alga
Enteromorpha
None
Trickle-flow (< 100 bubbles/min.)
Natural seawater or modified GP2, Forty Fathoms*
or equivalent, artificial seawater prepared using
Millipore MILLI-Q* or equivalent or deionized
water
Site sediment, a reference sediment and a control
sediment
N/A
Survival
<6 wk
2L
i 90% survival in controls
ASTM.  1991.  E1367-90.  Standard guide for conducting 10-day static sediment toricity tests with marine
    and estuarine amphipods.  Annual Book of ASTM Standards, Vol. 11.04.  American Society for
    Testing and Materials, Philadelphia, PA.
*   Static renewal, intermittent flow or continuous flow tests may be used where it is necessary to
    maintain water quality parameters, e.g., dissolved oxygen (DO). For static renewal tests the overlying
    dilution water should be changed every 48 h at a minimum.

-------
                                             E-21
       SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE
                AMPHIPOD, Corophium sp., ACUTE TOXICITY SEDIMENT TESTS
  1.  Test type:
  2.  Test duration:
  3.  Temperature:
  4.  Salinity:
  5.  Light quality:
  6.  Light intensity:
  7.  Photoperiod:
  8.  Test chamber size:
  9.  Test solution volume:
  10. Sediment depth:
  11. Renewal of test solutions:
  12. Age of test organisms:
  13. No. of organisms per test chamber:
  14. No. replicate chambers per sediment:
  IS. No. organisms per sediment:
  16. Feeding regime:
  17. Test chamber cleaning:
  18. Test solution aeration:
  19. Dilution water:
 20. Test concentrations:

 21. Dilution series:
 22. Endpoint:
 23. Sampling and sample holding requirements:
 24. Sample volume required:
 25. Test acceptability criterion:
Static Non-renewal*
10 d
15-25°C
Variable, species dependent
Ambient Laboratory
10-20 uE/m2/s (50-100 ft-c)
Continuous Light
1L
Vol. to 950 mL
2 cm minimum
None*
Mature 5 - 8 mm amphipods, mixed sexes
20
5
100
None
None
Trickle-flow (<  100 bubbles/min.)
Natural seawater or modified GP2, Forty
Fathoms* or equivalent, artificial seawater
prepared with Millipore MILLI-Q or equivalent or
deionized water
Site sediment, a reference sediment and a control
sediment
N/A
Survival
<6 wk
2L
* 90% survival in controls
REFERENCE:
ASTM.  1991. E1367-90.  Standard guide for conducting 10-day static sediment toxicity tests with marine
   and estuarine amphipods. Annual Book of ASTM Standards, vol. 11.04.  American Society for Testing
   and Materials, Philadelphia, PA.
*  Static renewal, intermittent flow or continuous flow tests may be used where it is necessary to maintain
   water quality parameters, e.g., dissolved oxygen (DO). For static renewal tests the overlying dilution
   water should be changed every 48 h at a minimum.

-------
                                             E-22
      SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE
             AMPHIPOD, Eohaustorius estuarius, ACUTE TOXICITY SEDIMENT TESTS
 1.  Test type:
 2.  Test duration:
 3.  Temperature:
 4.  Salinity:
 5.  Light quality:
 6.  Light intensity:
 7.  Photoperiod:
 8.  Test chamber size:
 9.  Test solution volume:
 10. Sediment depth:
 11. Renewal of test solutions:
 12. Age of test organisms:
 13. No. of organisms per test chamber:
 14. No. replicate chambers per sediment:
 15. No. organisms per sediment:
 16. Feeding regime:
 17. Test chamber cleaning:
 18. Test solution aeration:
 19. Dilution water:
 20.  Test concentrations:

 21.  Dilution series:
 22.  Endpoint:
 23.  Sampling and sample holding requirements:
 24.  Sample volume required:
 25.  Test acceptability criterion:
Static Non-renewal*
10 d
15±3°C
2 to s28 %o
Ambient Laboratory
10-20 uE/m2/s (50-100 ft-c)
Continuous Light
1L
Vol. to 950 mL
2 cm minimum
None*
Mature amphipods, 3 -5 mm, mixed sexes
20
5
100
None
None
Trickle-flow (< 100 bubbles/min.)
Natural Seawater or modified GP2, Forty Fathoms*
or equivalent, artificial seawater prepared using
Millipore MILLI-Q* or equivalent or deionized
water
Site sediment, a reference sediment and a control
sediment
N/A
Survival
<6wk
2L
* 90% survival in  controls
REFERENCE:
ASTM.  1991.  E1367-90.  Standard guide for conducting 10-day static sediment toxicity tests with marine
    and estuarine amphipods. Annual Book of ASTM Standards, Vol. 11.04. American Society for
    Testing and Materials, Philadelphia, PA
*   Static renewal, intermittent flow or continuous flow tests may be used where it is necessary to
    maintain water quality parameters, e.g., dissolved oxygen (DO). For static renewal tests the overlying
    dilution water should be changed every 48 h at a minimum.

-------
                                             E-23
  SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE MAYFLY,
                     Hexagenia timbata, ACUTE TOXICITY SEDIMENT TESTS
 1.  Test type:
 2.  Test duration:
 3.  Temperature:
 4.  Salinity:
 5.  Light quality:
 6.  Light intensity:
 7.  Photoperiod:
 8.  Test chamber size:*
 9.  Test solution volume:*
 10. Sediment depth:
 11. Renewal of test solutions:
 12. Age of test organisms:
 13. No. organisms per test chamber:
 14. No. replicate chambers per concentrations:
 IS. No. organisms per concentration:
 16. Feeding regime:*
 17. Test chamber cleaning:
 18. Test solution aeration:
 19. Dilution water:
 20. Test concentrations:

 21. Dilution series:
 22. Endpoint:
 23. Sampling and sample holding requirements:
 24. Sample volume required:
 25. Test acceptability:
* - Protocol Dependent
Static Non-renewal*
10 d
17°C, 20-22°C
0%o
Ambient Laboratory
10-20 uE/mVs (50-100 ft-c)
16L/8D
1L
Vol. to 800 mL
2 cm minimum
None*
young nymphs
5 minimum
4 minimum
1-10
Variable
None
Trickle-flow (< 100 bubbles/min.)
Moderately hard synthetic water prepared using
Millipore MILLI-Q* or equivalent deionized water
and reagent grade chemicals or 20% DMW, receiving
water, or synthetic water modified to reflect receiving
water hardness
Site sediment, a reference sediment and a control
sediment
None
Survival
<6wk
2L
* 80% survival in  controls
REFERENCE:
Bedard, D., A. Hayton and D. Persaud.  1992.  Ontario Ministry of the Environment laboratory sediment
    biological testing protocol.  Ontario Ministry of the Environment, Toronto, Ontario. 26 pp.
*   Static renewal, intermittent flow or continuous flow tests may be used where it is necessary to
    maintain water quality parameters, e.g., dissolved oxygen (DO). For static renewal tests the overlying
    dilution water should be changed every 48 h at a  minimum.

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                                           E-24
   SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE
      FRESHWATER AMPHIPOD, HyaleUa azteca, ACUTE TOXICITY SEDIMENT TESTS
1.   Test type:
2.   Test duration:
3.   Temperature:
4.   Salinity
5.   Light quality:
6.   Light intensity:
7.   Photoperiod:
8.   Test chamber size:
9.   Test solution volume:
10.  Sediment depth:
11.  Renewal of test solutions:
12.  Age of test organisms:
13.  No. organisms per test chamber:
14.  No. replicate chambers per sediment:
15.  No. organisms per sediment:
16.  Feeding regime:

17.  Test chamber cleaning:
18.  Test solution aeration:
19.  Dilution water:
20.  Test concentrations:

21.  Dilution series:
22.  Endpoint:
23.  Sampling and sample holding requirements:
24.  Sample volume required:
25.  Test acceptability criterion:
Static Non-renewal*
10 d
20 - 25°C
0-15 %o
Ambient Laboratory
10-20 uE/m2/s (50-100 ft-c)
16L/8D
300 mL minimum
Variable, depending on test type
2 cm minimum
None*
7- 14 d
10 minimum
5 minimum
50 minimum
Variable (None, Tetrafm, YCT*, rabbit chow,
maple leaves)
None
Trickle-flow (<100 bubbles/min.)
Moderately hard synthetic water prepared using
Millipore MILLI-Q* or equivalent deionized
water and reagent  grade chemicals or 20%
DMW, receiving water, or synthetic water
modified to reflect receiving water hardness
Site sediment, a reference sediment and a
control sediment
N/A
Survival
<6wk
2L
i 80% survival in controls
   Slurry of Yeast, Cereal flakes, Trout chow

-------
                                              E-25

REFERENCES:

ASTM.  1991.  Standard guide for conducting sediment toxicity tests with freshwater invertebrates.
    Method E1383-90.  Annual Book of ASTM Standards, Vol. 11.04.  American Society for Testing and
    Materials, Philadelphia, PA

Ingersoll, C.G. and M.K. Nelson.  1990.  Testing sediment toxicity with Hyalella azteca (Amphipoda) and
    Chironomus riparius (Diptera). pp. 93-109  In W.G. Landis and W.H. van der Schalie, eds., Aquatic
    Toxicology and Risk Assessment:  Thirteenth Volume.  ASTM STP 109b.  American Society for
    Testing and materials, Philadelphia, PA

*   Static renewal, intermittent flow or continuous flow tests may be used where it is necessary to
    maintain water quality parameters, e.g., dissolved oxygen (DO). For static renewal tests the overlying
    dilution water should be changed every 48 h at a minimum.

-------
                                             E-26
       SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE
           POLYCHAETE, Neanthes arenaceodentata, ACUTE TOXICITY SEDIMENT TESTS
 1.   Test type:
 2.   Test duration:
 3.   Temperature:
 4.   Salinity:
 5.   Light quality:
 6.   Light intensity:
 7.   Photoperiod:
 8.   Test chamber size:
 9.   Test solution volume:
 10.  Sediment depth:
 11.  Renewal of test solutions:
 12.  Age of test organisms:
 13.  No. organisms per test chamber:
 14.  No. replicate chambers per concentration:
 IS.  No. organisms per concentration:
 16.  Feeding regime:
 17.  Test chamber cleaning:
 18.  Test solution aeration:
 19.  Dilution water:

 20.  Test concentrations:

 21.  Dilution series:
 22.  Endpoint:
 23.  Sampling and sample holding requirements:
 24.  Sample volume required:
 25.  Test acceptability criterion:
REFERENCES:
Static Non-renewal*
10 d
20±1°C
20-35 %o
Ambient Laboratory
10-20 uE/m2/fc (50-100 ft-c)
12L/12D
1L
Vol. to 800 mL
2.5 cm (200 mL)
None*
2-3 weeks
5 maximum
3-5
15-25
None
None
Trickle-flow (< 100 bubbles/min.)
Natural  seawater or modified GP2, Forty Fathoms*,
or equivalent, artificial seawater prepared with
Millipore MILLI-Q* or eqivalent or deionized water
Site sediment, a reference sediment and a control
sediment
N/A
Survival
<6 wk
2L
i 90% survival in controls
Dillon, T.M., D.W. Moore and A.B. Gibson. 1993.  Development of a chronic sublethal bioassay for
    evaluating contaminated sediment with the marine polychaete worm, Nereis (Neanthes)
    arenaceodentata.  Environ. Toxicol. Chem. 12:589-605.
Reish, D.J. 1992.  Guide for conducting sediment toxicity tests with marine and estuarine polychaetous
    annelids.  ASTM Draft No. 5.  July 3, 1992.  American Society for Testing and Materials, Philadelphia,
    PA
*   Static renewal, intermittent flow or continuous flow tests may be used where it is necessary to
    maintain water quality parameters, e.g., dissolved oxygen (DO). For static renewal tests the overlying
    dilution water should be changed every 48 h at a minimum.

-------
                                             E-27
   SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE PAPER
           PONDSHELL CLAM, Anodonla imbecillis, ACUTE TOXICITY SEDIMENT TESTS
 1.  Test type:
 2.  Test duration:
 3.  Temperature:
 4.  Salinity:
 5.  Light quality:
 6.  Light intensity:
 7.  Photoperiod:
 8.  Test chamber size:

 9.  Test solution volume:
 10. Sediment depth:
 11. Renewal of test solutions:
 12. Age of test organisms:
 13. No. organisms per test chamber:
 14. No. replicate chambers per concentration:
 IS. No. organisms per concentration:
 16. Feeding regime:
 17. Test chamber cleaning:
 18. Test solution aeration:
 19. Dilution water:
 20. Test concentrations:

 21. Dilution series:
 22. Endpoint:

 23. Sampling and sample holding requirements:
 24. Sample volume required:
 25. Test acceptability criterion:
REFERENCE:
Static Non-renewal*
10 d
24±1°C
0%o
N/A
N/A
24 h Dark
5 cm-diam. glass cylinder closed on lower end
with 100 foa Nitex, placed in 250 mL glass dish
containing test sediment and overlying water
150 mL overlying water
0.5 cm (20 mL)
None*
8-10 d post transformation to juveniles
10
5 minimum
50 minimum
None
None
None
Moderately hard synthetic water prepared using
Millipore MILLI-Q* or equivalent deionized
water  and reagent grade chemicals or 20%
DMW, receiving water or filtered non-toxic
natural freshwater
Site sediment, a reference sediment and a control
sediment
N/A
Survival (death assumed if absence of ciliary
action or empty shells)
<6 wk
2L
i 80% survival in controls
Tennessee Valley Authority Draft Standard Operating Procedures, SOP-21, and personal communication
    from Don Wade, Tennessee Valley Authority.
*   Static renewal, intermittent flow or continuous flow tests may be used where it is necessary to
    maintain water quality parameters, e.g., dissolved oxygen (DO). For static renewal tests the overlying
    dilution water should be changed every 48 h at a minimum.

-------
                                            E-28
     SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR MYSID
  SHRIMP, Mysidopsis bahia, M. bigelowi, M. almyra, Neomysis americana, Holmesimysis costata, ACUTE
                                TOHCITY SEDIMENT TESTS
1.   Test type:
2.   Test duration:
3.   Temperature:
4.   Salinity:

5.   Light quality:
6.   Light intensity:
7.   Photoperiod:
8.   Test chamber size:
9.   Test solution volume:
10. Sediment depth:
11. Renewal of test solutions:
12. Age of test organisms:
13. No. organisms per test chamber:
14. No. replicate chambers per concentration:
15. No. organisms per concentration:
16. Feeding regime:
17. Test chamber cleaning:
18. Test solution aeration:
19. Dilution water:


20. Test concentrations:

21. Dilution series:
Static Non-renewal*
10 d
20±1°C: or 25±1°C for
  Mysidopsis bahia
  Mysidopsis bigelowi
  Mysidopsis almyra
20±1°C for
  Neomysis americana
12±l°Cfor
  Holmesimysis costata
25-30 %o ±10% except for Holmesimysis costata which
is to be 32-34 %o ±10%
Ambient Laboratory
10-20 uE/mVs (50-100 ft-c)
16L/8D
250 mL (minimum)
200 mL (minimum)
2 cm minimum
None*
1 - 5 d; 24 h range  in age
10 minimum
5 minimum
50 minimum
Anemia nauplii are made available while holding
prior to, but not during, the test; feed 0.2 mL of
concentrated suspension of Anemia nauplii s24 h
old,  daily (approximately 100 nauplii per mysid)
None
If needed to maintain DO> 40% saturation for:
 Mysidopsis bahia
 Mysidopsis bigelowi
 Mysidopsis almyra
 Neomysis americana
and  DO> 60% saturation for:
 Holmesimysis costata
(< 100 bubbles/min.)
Natural seawater or modified GP2, Forty Fathoms*
or equivalent, artificial seawater prepared with
Millipore MILLI-Q* or equivalent or deionized
water
Site  sediment, a reference sediment and a control
sediment
N/A

-------
                                              E-29


 22. Endpoint:                                  Survival

 23. Sampling and sample holding requirements:   <6 wk

 24. Sample volume required:                    1 L

 25. Test acceptability criterion:                  * 90% survival in controls
REFERENCE:

Modified from:

USEPA.  1991.  Methods for Measuring the Acute Toricity of Effluents and Receiving Waters to
    Freshwater and Marine Organisms, 4th Ed. EPA/600/4-90/027.

*   Static renewal, intermittent flow or continuous flow tests may be used where it is necessary to
    maintain water quality parameters, e.g., dissolved oxygen (DO). For static renewal tests the overlying
    dilution water should be changed every 48 h at a minimum.

-------
                                             E-30
  SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR COMMERCIAL
                   SHRIMP, Penaeus sp., ACUTE TOXICITY SEDIMENT TESTS
 1.  Test type:
 2.  Test duration:
 3.  Temperature:
 4.  Salinity:
 5.  Light quality:
 6.  Light intensity:
 7.  Photoperiod:
 8.  Test chamber size:
 9.  Test solution volume:

 10. Sediment depth:
 11. Renewal of test solutions:
 12. Age of test organisms:
 13. No. organisms per test chamber:
 14. No. replicate chambers per concentration:
 IS. No. organisms per concentration:
 16. Feeding regime:
 17. Test chamber cleaning:
 18. Test solution aeration:

 19. Dilution water:
 20. Test concentrations:

 21. Dilution series:
 22. Endpoint:
 25. Test acceptability criterion:
REFERENCE:
Static Non-renewal*
10 d
25±1°C
30-35 %o± 10%
Ambient Laboratory
10-20 uE/mVs (50-100 ft-c)
16L/8D
80 L minimum
60 L minimum; overlying water variable depending
on test type
2 cm minimum
None*
8-10 d post larvae
10 minimum
5 minimum
50 minimum
None
None
If needed to maintain DO> 40% saturation
(< 100 bubbles/min.)
Natural seawater or modified GP2, Forty Fathoms*
or equivalent, artifical seawater prepared with
Millipore MILLI-Q* or equivalent or deionized
water
Site sediment, a reference sediment and a control
sediment
N/A
Survival
 23. Sampling and sample holding requirements:   <6 wk
 24. Sample volume required:
20 L for site sediment and 8 L for reference and
control sediment
* 80% survival in controls
Modified from the mysid acute toxicity water column test published in:
USEPA.  1991.  Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to
    Freshwater and Marine Organisms, 4th Ed.  EPA/600/4-90/027.
*   Static renewal, intermittent flow or continuous flow tests may be used where it is necessary to
    maintain water quality parameters, e.g., dissolved oxygen (DO). For static renewal tests the overlying
    dilution water should be changed every 48 h at a minimum.

-------
                                             E-31
      SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR GRASS
                 SHRIMP, Palaemonetes sp., ACUTE TOXICITY SEDIMENT TESTS
 1.  Test type:
 2.  Test duration:
 3.  Temperature:
 4.  Salinity:
 5.  Light quality:
 6.  Light intensity:
 7.  Photoperiod:
 8.  Test chamber size:
 9.  Test solution volume:

 10. Sediment depth:
 11. Renewal of test solutions:
 12. Age of test organisms:
 13. No. organisms per test chamber:
 14. No. replicate chambers per concentration:
 15. No. organisms per concentration:
 16. Feeding regime:
 17. Test chamber cleaning:
 18. Test solution aeration:

 19. Dilution water:
 20. Test concentrations:

 21. Dilution series:
 22. Endpoint:
 23. Sampling and sample holding requirements:
 24. Sample volume required:
Static Non-renewal*
10 d
25±1°C
2 %o to i28 %o
Ambient Laboratory
10-20 uE/mVs (50-100 ft-c)
16L/8D
80 L minimum
60 L minimum; overlying water variable depending
on test type
2 cm minimum
None*
1-4 d from hatch
10 minimum
5 minimum
50 minimum
None
None
If needed to maintain DO > 40% saturation
(<  100 bubbles/min.)
Natural seawater or modified GP2, Forty Fathoms*
or equivalent, artificial seawater prepared with
Millipore MILLI-Q* or equivalent or deionized
water
Site sediment, a reference sediment and a control
sediment
N/A
Survival
<6 wk
20 L for site sediment and 8 L for reference and
control sediment
i 80% survival in controls
 25. Test acceptability criterion:
REFERENCE:
Modified from the mysid acute toxicity water column test published in:
USEPA.  1991.  Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to
    Freshwater and Marine Organisms, 4th Ed.  EPA/600/4-90/027.

-------
                                            E-32

Static renewal, intermittent flow or continuous flow tests may be used where it is necessary to
maintain water quality parameters, e.g., dissolved oxygen (DO). For static renewal tests the overlying
dilution water should be changed every 48 h at a minimum.

-------
                                             E-33
     SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR MIDGES,
              Chironomus tentans AND C. riparius, ACUTE TOXICITY SEDIMENT TESTS
  1.  Test type:
  2.  Test duration:
  3.  Temperature:
  4.  Salinity:
  5.  Light quality:
  6.  Light intensity:
  7.  Photoperiod:
  8.  Test chamber size:
  9.  Test solution volume:

  10. Sediment depth:
  11. Renewal of test solutions:
  12. Age of test organisms:
  13. No. organisms per test chamber:
  14. No. replicate chambers per concentration:
  IS. No. organisms per concentration:
  16. Feeding regime:
  17. Test chamber cleaning:
  18. Test solution aeration:
  19. Dilution water:
  20. Test concentrations:

  21. Dilution series:
  22. Endpoint:
  23. Sampling and sample holding requirements:
  24. Sample volume required:
  25. Test acceptability criterion:
*   Slurry of Yeast, Cereal flakes, Trout chow.
REFERENCES:
Ingersoll, C.G. and M.K. Nelson. 1990. Testing sediment toxicity with Hyalella azteca (Amphipoda) and
    Chironomus riparius (Diptera).  pp. 93-109. In W.G. Landis and W.H. van der Schalie, eds., Aquatic
    Toxicology and Risk Assessment: Thirteenth Volume.  ASTM STP 109b.  American Society for
    Testing and  Materials, Philadelphia, PA
ASTM.  1991. New standard guide for conducting solid-phase sediment toxicity tests with freshwater
    invertebrates. ASTM Draft Document E1383. American Society for Testing and Materials,
    Philadelphia, PA
Static Non-renewal*
10 d
20 or 25°C
0%o
Ambient Laboratory
10-20 uE/m2/s (50-100 ft-c)
16U8D
300 mL minimum
100 mL sediment minimum; overlying water variable
depending on test type
2 cm minimum
None*
1st - 2nd Instar
10 minimum
5 minimum
50 minimum
Variable (None, Tetramin, YCT*)
None
Trickle-flow (<  100 bubbles/min.)
Variable
Site sediment, a reference sediment and a control
sediment
N/A
Survival
<6 wk
4L
^ 70% survival in controls

-------
                                           E-34

Static renewal, intermittent flow or continuous flow tests may be used where it is necessary to
maintain water quality parameters, e.g., dissolved oxygen (DO). For static renewal tests the overlying
dilution water should be changed every 48 h at a minimum.

-------
                                             E-35
   SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE NAIDID
               OLIGOCHAETE, Pristina leidyi, ACUTE TOXICITY SEDIMENT TESTS
  1.  Test type:
  2.  Test duration:
  3.  Temperature:
  4.  Salinity:
  5.  Light quality:
  6.  Light intensity:
  7.  Photoperiod:
  8.  Test chamber size:
  9.  Test solution volume:
  10. Sediment depth:
  11. Renewal of test solutions:
  12. Age of test organisms:
  13. No. of organisms per test chamber:
  14. No. replicate chambers per concentration:
  IS. No. organisms per concentration:
  16. Feeding regime:
  17. Test chamber cleaning:
  18. Test solution aeration:
  19. Dilution water:
  20. Test concentrations:

  21. Dilution series:
  22. Endpoint:
  23. Sampling and sample holding requirements:
  24. Sample volume required:
  25. Test acceptability criterion:
Static Non-renewal*
10 d
24±1°C
0%o
Ambient Laboratory
10-20 uE/m2/s (50-100 ft-c)
16L:8D
250 mL
10 g (wet wt)/50 mL overlying water
2 cm minimum
None*
Mixed age
5
5
25
None
None
None
Variable
Site sediment, a reference sediment and a control
sediment
N/A
Survival
<6 wk
500 mL
i 90% survival in controls
REFERENCES:
Smith, D.P., J.H. Kennedy and K.L. Dickson. 1991. An evaluation of a naidid oligochaete as a toxicity test
    organism. Environ. Toxicol. Chem. 10: 1459-1465.
*   Static renewal, intermittent flow or continuous flow tests may be used where it is necessary to
    maintain water quality parameters, e.g., dissolved oxygen (DO). For static renewal tests the overlying
    dilution water should be changed every 48 h at a minimum.

-------
                                             E-36
     SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE
             OLIGOCHAETE, Tubifex tubtfex, ACUTE TOXICITY SEDIMENT TESTS
 1.  Test type:
 2.  Test duration:
 3.  Temperature:
 4.  Salinity:
 5.  Light quality:
 6.  Light intensity:
 7.  Photoperiod:
 8.  Test chamber size:
 9.  Test solution volume:
 10. Sediment depth:
 11 Renewal of test solutions:
 12. Age of test organisms:
 13. No. organisms per test chamber:
 14. No. replicate chambers per sediment:
 15. No. organisms per sediment:
 16. Feeding regime:
 17. Test chamber cleaning:
 18. Test solution aeration:
 19. Dilution water:
Static Non-renewal*
10 d
20 - 25°C
0%o
Ambient Laboratory
10-20 uE/m2/s (50-100 ft-c)
16L/8D
250 mL
100 mL
100 mL
None*
Mixed age
5
5
25
None
None
None
Moderately hard synthetic water prepared using
Millipore MILLI-Q* or equivalent, deionized
water and reagent grade chemicals or 20% DMW,
receiving water, or synthetic water modified to
reflect receiving water hardness
Site sediment, a reference sediment and a control
sediment
N/A
Survival
<6wk
1L
* 90% survival in controls
 20. Test concentrations:

 21. Dilution series:
 22. Endpoint:
 23. Sampling and sample holding requirements:
 24. Sample volume required:
 25. Test acceptability criterion:
REFERENCE:
Adapted from:
Reynoldson, T.B., S.P. Thompson and J.L. Bamsey. 1991. A sediment bioassay using the tubified
    oligochaete worm Tubifex tubifex. Environ. Toxicol. Chem. 10:1061-1072.
*   Static renewal, intermittent flow or continuous flow tests may be used where it is necessary to
    maintain water quality parameters, e.g., dissolved oxygen  (DO). For static renewal tests the overlying
    dilution water should be changed every 48 h at a minimum.

-------
                                            E-37
     SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE
         OLIGOCHAETE, Lumbriculus variegatus, ACUTE TOX1CITY SEDIMENT TESTS
1.  Test type:
2.  Test duration:
3.  Temperature:
4.  Salinity:
5.  Light quality:
6.  Light intensity:
7.  Photoperiod:
8.  Test chamber size:
9.  Test solution volume:
10. Sediment depth:
11  Renewal of test solutions:
12. Age of test organisms:
13. No. organisms per test chamber:
14. No. replicate chambers per sediment:
15. No. organisms per sediment:
16. Feeding regime:
17. Test chamber cleaning:
18. Test solution aeration:
19. Dilution water:
20.  Test concentrations:

21.  Dilution series:
22.  Endpoint:
23.  Sampling and sample holding requirements:
24.  Sample volume required:
25.  Test acceptability criterion:
Static Non-renewal*
10 d
20 - 25°C
0%o
Ambient Laboratory
10-20 uE/m2As (50-100 ft-c)
16L/8D
300 mL minimum
100 mL minimum
3 cm
None*
Mixed age
10
5
50
10 mg trout chow starter on days 0, 5
None
None
Moderately hard synthetic water prepared using
Millipore MILLI-Q* or equivalent, deionized
water and reagent grade chemicals or 20% DMW,
receiving water, or synthetic water modified to
reflect receiving water hardness
Site sediment, a reference sediment and a control
sediment
N/A
Survival
<6 wk
1L
i 90% survival in controls

-------
                                              E-38

REFERENCES:

Adapted from:

Ankley, G.T., R.A. Hoke, D.A. Benoit, E.N. Leonard, C.W. West, G.L. Phipps, V.R. Mattson and L.A.
    Anderson. 1993.  Development and evaluation of test methods for benthic invertebrates and sediments:
    effects of flow rate and feeding on water quality and exposure conditions. Arch. Environ. Contain.
    Toxicol. 25:12-19.

Bailey, N.C and D.N.W. Lui. 1980. Lumbriculus variegatus, a benthic oligochaete, as a bioassay organism.
    Pp. 202-215. In: J.C. Eaton, P.R. Parrish and AC. Hendricks (Eds). Aquatic Toxicology. ASTM STP
    707. American Society for Testing and Materials, Philadelphia, PA.

*   Static renewal, intermittent flow or continuous flow tests may be used where it is necessary to
    maintain water quality parameters, e.g., dissolved oxygen  (DO). For static renewal tests the overlying
    dilution water should be changed every 48 h at a minimum.

-------
      Sediment
Bioaccumulation Tests
                                DRAFT

-------
                                             E-40
      SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE
         POLYCHAETE, Neanthes arenaceodentata, SEDIMENT BIOACCUMULATION TESTS
 1.  Test type:
 2.  Test duration:
 3.  Temperature:
 4.  Salinity:
 5.  Light quality:
 6.  Light intensity:
 7.  Photoperiod:
 8.  Test chamber size:
 9.  Test solution volume:
 10. Sediment depth:
 11. Renewal of test solutions:
 12. Age of test organisms:
 13. No. organisms per test chamber:
 14. No. replicate chambers per concentration:
 IS. No. organisms per concentration:
 16. Feeding regime:
 17. Test chamber cleaning:
 18. Test solution aeration:
 19. Dilution water:
 20.  Test concentrations:

 21.  Dilution series:
 22.  Endpoint:
 23.  Sampling and sample holding requirements:
 24.  Sample volume required:
 25.  Test acceptability criterion:
REFERENCES:
Static Renewal
28 d
20±1°C
20-35 %o
Ambient Laboratory
10-20 uE/m2/s (50-100 ft-c)
12L/12D
1 L minimum
200 mL overlying water
2.5 cm (200 mL)
Weekly
2-3 wk
5 maximum
5 minimum
25 minimum
None
None
Trickle-flow (<  100 bubbles/min.)
Natural seawater or modified GP2, Forty Fathoms*,
or equivalent, artificial seawater prepared with
Millipore MILLI-Q*, or eqivalent or deionized
water
Site sediment, a reference sediment and a control
sediment
N/A
Bioaccumulation
< 6wk
8L
Adequate mass of organisms at test completion for
detection of target analyte(s)
Dillon, T.M., D.W. Moore and AB. Gibson. 1993. Development of a chronic sublethal bioassay for
    evaluating contaminated sediment with the marine polychaete worm, Nereis (Neanthes)
    arenaceodentata. Environ. Toricol. Chem. 12:589-605.
Reish, D.J. 1992.  Guide for conducting sediment toxicity tests with marine and estuarine polychaetous
    annelids. ASTM Draft No. 5. July  3,1992. American Society for Testing and Materials, Philadelphia,
    PA

-------
                                             E-41
       SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE
               POLYCHAETE, Nereis virens, SEDIMENT BIOACCUMULATION TESTS
 1.  Test type:
 2.  Test duration:
 3.  Temperature:
 4.  Salinity:
 5.  Light quality:
 6.  Light intensity:
 7.  Photoperiod:
 8.  Test chamber size:

 9.  Test solution volume:
 10. Sediment depth:
 11. Renewal of test solutions:

 12. Age of test organisms:
 13. No. organisms per test chamber:
 14. No. replicate chambers per sediment:
 15. No. organisms per sediment:
 16. Feeding regime:
 17. Test chamber cleaning:
 18. Test solution aeration:
 19. Dilution water:
 20. Test concentrations:

 21. Dilution series:
 22. Endpoint:
 23. Sampling and sample holding requirements:
 24. Sample volume required:
 25. Test acceptability criterion:

REFERENCE:
Flow-through or Static Renewal
28 d
10 to 20°C
*20%o
Ambient Laboratory
10-20 uE/mz/s (50-100 ft-c)
16L/8D, 14L/10D, 12L/12D
1 L (beaker) or large chamber with multiple worms
composited into a single replicate (e.g., 20 worms in
20 gallon aquarium)
> 750 mLAvorm
* 4cm
Flow-through = 5-10 vol/d; Static Renewal =
3x/week
adult (3 - 15g)
One per 1 L beaker, 20 per 20 gallon aquarium
5-8 (depending on desired statistical power)
5-8 (assumes values to be determined on individuals)
None
As needed
Moderate, as needed
Natural seawater or modified GP, Forty Fathoms*
or equivalent, artificial seawater prepared with
Millipore MILLI-Q* or equivalent or deionized
water
Site sediment, a reference sediment and control
sediment
N/A
Bioaccumulation
< 6 wk
200 mL per worm
Adequate mass of organisms at test completion for
detection of target analyte(s)
Lee II, H., B. Boese, J. Pelletier, M. Winsor, D. Specht and R. Randall. 1989. Guidance Manual: Bedded
    Sediment Bioaccumulation Tests. EPA/600/X-89/302. U.S. Environmental Protection Agency. 232 pp.

-------
                                            E-42
      SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE
            POLYCHAETE, Arenicola marina, SEDIMENT BIOACCUMULATION TESTS
1.   Test type:
2.   Test duration:
3.   Temperature:
4.   Salinity:
5.   Light quality:
6.   Light intensity:
7.   Photoperiod:
8.   Test chamber size:
9.   Test solution volume:

10. Sediment depth:

11. Renewal of test solutions:

12. Age of test organisms:

13. No. organisms per test chamber:
14. No. replicate chambers per sediment:
IS. No. organisms per sediment:
16. Feeding regime:
17. Test chamber cleaning:
18. Test solution aeration:
19. Dilution water:
20. Test concentrations:

21. Dilution series:
22. Endpoint:
23. Sampling and sample holding requirements:
24. Sample volume required:
25. Test acceptability criterion:
Flow-through or Static Renewal
28 d
10 to 20°C
Ambient Laboratory
10-20 uE/m2/s (50-100 ft-c)
16L/8D, 14L/10D, 12L/12D
1-2 L
> 500 mL/beaker (e.g., four 1 L beakers in 8 L
aquarium)
* 15 cm deep sediment (wet wt); minimum 400 g
sediment (wet wt) plus 3.5 g sediment per g wet flesh
weight per day (s 250 mm in grain size diameter)
Flow-through = 5-10 vol/d; Static Renewal =
3x/week
< 1 year (3-6 g wet weight, 5-10 cm length), larger
organisms require more sediment, larger test
chambers
One (1) per beaker maximum
5-8 (depending on desired statistical power)
5-8 (assumes values to be determined on individuals)
None
As needed
Moderate, as needed
Natural seawater or modified GP, Forty Fathoms*
or equivalent, artificial seawater prepared with
Millipore MILLI-Q* or equivalent or deionized
water
Site sediment, a reference sediment and control
sediment
N/A
Bioaccumulation
< 6 wk
1 L per treatment, minimum
Adequate mass of organisms at test completion for
detection of target analyte(s)

-------
                                              E-43

REFERENCES:

Lee II, H., B. Boese, J. Pelletier, M. Winsor, D. Specht and R. Randall. 1989. Guidance Manual: Bedded
    Sediment Bioaccumulation Tests. EPA/600/X-89/302. U.S. Environmental Protection Agency. 232 pp.

Gordon, D.C., J. Dale and P.D. Keiger. 1978. Importance of sediment-working by the deposit-feeding
    polychaete Arenicola marine on the weathering rate of sediment-bound oil. J. Fish Res. Bd. Canada.
    35:591-603.

Huttel, M. 1990. Influence of the lugworm Arenicola marina on porewater nutrient profiles of sand flat
    sediments. Mar. Biol. Prog. Ser. 62:241-248.

-------
                                            E-44
     SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE
       OLJGOCHAETE, Lumbriculus variegates, SEDIMENT BIOACCUMULATION TESTS
1.   Test type:
2.   Test duration:
3.   Temperature:
4.   Salinity:
5.   Light quality:
6.   Light intensity:
7.   Photoperiod:
8.   Test chamber size:
9.   Test solution volume:
10.  Sediment depth:
11  Renewal of test solutions:
12.  Age of test organisms:
13.  No. organisms per test chamber:
14.  No. replicate chambers per sediment:
IS.  No. organisms per sediment:
16,  Feeding regime:
17.  Test chamber cleaning:
18.  Test solution aeration:

19.  Dilution water:
20. Test concentrations:

21. Dilution series:
22. Endpoint:
23. Sampling and sample holding requirements:
24. Sample volume required:
25. Test acceptability criterion:
Static Non-renewal or Overlying Water Renewal
28 d
20-25°C
0%o
Ambient Laboratory
10-20 uE/mVs (50-100 ft-c)
16L/8D
4 L minimum
1L
3cm
Variable
Mixed Age Adults
5 g (-500-1000) (Minimum)
4 minimum
N/A
None
None
If needed to maintain DO> 40% saturation
(<  100 bubbles/min.)
Moderately hard synthetic water prepared using
Millipore MILLI-Q* or equivalent, deionized
water and reagent grade chemicals or 20% DMW,
receiving water, or synthetic water modified to
reflect receiving water hardness
Site sediment, a reference sediment and a control
sediment
N/A
Bioaccumulation
<6 wk
4L
Adequate mass of organisms at test completion for
detection of target analyte(s)

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                                              E-45

REFERENCES:

Ankley, G.T., R.A. Hoke, D.A. Benoit, E.N. Leonard, C.W. West, G.L. Phipps, V.R. Mattson and L.A.
    Anderson. 1993.  Development and evaluation of test methods for benthic invertebrates and sediments:
    effects of flow rate and feeding on water quality and exposure conditions.  Arch. Environ. Contain.
    Toricol. 25:12-19.

Phipps, G.L., G.T. Ankley, D.A. Benoit and V.R. Mattson.  1993. Use of the aquatic oligochaete
    Lumbriculus variegatus for assessing the toxicity and bioaccumulation of sediment-associated
    contaminants.  Environ. Toricol. Chem. 12:269-279.

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                                           E-46
SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE MACOMA
               CLAM, Macoma nasuta, SEDIMENT BIOACCUMULATION TESTS
1.   Test type:
2.   Test duration:
3.   Temperature:
4.   Salinity:
5.   Light quality:
6.   Light intensity:
7.   Photoperiod:
8.   Test chamber size:
9.   Test solution volume:

10.  Sediment depth:
11.  Renewal of test solutions:
12.  Age of test organisms:
13.  No. organisms per test chamber:
14.  No. replicate chambers per sediment.:
15.  No. organisms per sediment:
16.  Feeding regime:
17.  Test chamber cleaning:
18.  Test solution aeration:
19.  Dilution water:

20.  Test concentrations:

21.  Dilution series:
22.  Endpoint:
23.  Sampling and sample holding
    requirements:
24.  Sample volume required:
25.  Test acceptability criterion:
Flow-through or Static Renewal
28d
12 - 16°C
Ambient Laboratory
10-20 uE/m2/s (50-100 ft-c)
12L/12D, 16L/8D, 10L/14D
250mL - 1 L (beaker)
> 750 mL/beaker (e.g., ten 250 mL beakers in 8L
aquarium)
i 50 g wet wt sediment per g wet flesh (without shell)
Row-through = 5-10 vol/d; Static Renewal = 3 x/wk
2 - 4 yr, 28 - 45 mm shell length
One (1) per beaker maximum
5 - 8 (depending  on desired statistical power)
5-8 (assumes values to be determined on individuals)
None
As needed
Moderate, as needed
Natural seawater or modified GP2, Forty Fathoms*
or equivalent, artificial seawater prepared with
Millipore MILLI-Q* or equivalent or deionized water
Site sediment, a reference sediment and a control
sediment
N/A
Bioaccumulation
< 6 wk
8L
Adequate mass of organisms at test completion for
detection of target analyte(s)

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                                              E-47

REFERENCES:

Lee II, H., B. Boese, J. Pelletier, M. Winsor, D. Specht, and R. Randall. 1989. Guidance Manual: Bedded
    Sediment Bioaccumulation Tests. EPA/600/X-89/302. 232 pp.

Ferraro, S., H. Lee II, R. Ozretich, and D. Specht. 1990. Predicting bioaccumulation potential: A test of a
    fugacity-based model. Arch. Environ. Contamin. Toxicol. 19:386-394.

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                                            E-48
 SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE CLAM,
                   Yoldia timatula, SEDIMENT BIOACCUMULATION TESTS
1.   Test type:
2.   Test duration:
3.   Temperature:
4.   Salinity:
5.   Light quality:
6.   Light intensity:
7.   Photoperiod:
8.   Test chamber size:
9.   Test solution volume:
10.  Sediment depth:
11  Renewal of test solutions:

12.  Age of test organisms:
13.  No. organisms per test chamber:
14.  No. replicate chambers per sediment:
15.  No. organisms per sediment:

16.  Feeding regime:
17.  Test chamber cleaning:
18.  Test solution aeration:
19.  Dilution water:
20. Test concentrations:

21. Dilution series:
22. Endpoint:
23. Sampling and sample holding requirements:
24. Sample volume required:
25. Test acceptability criterion:
Flow-Through or Static Renewal
28 d
5 - 20°C (activity minimal at lowest temperature)
Ambient Laboratory
10-20 uE/m2/s (50-100 ft-c)
16U8D, 14L/10D, 12L/12D
500 - 1000 mL (beaker)
>750 mL/beaker
100 - 300 g sediment (dry wt), depth greater than
shell length. Yoldia actively resuspends sediments
into water column, additional sediment may need
to be added during test to maintain minimal
sediment depth.
Flow-through = 5-10 vol/d; Static Renewal =
3x/week
1 - 2 cm g
One (1) per beaker
5-8 (depending on desired statistical power)
5 - 8 (assumes values to be determined on
individuals)
None
As needed
Moderate, as needed
Natural seawater or modified GP, Forty Fathoms*
or equivalent, artificial seawater prepared with
Millipore MILLI-Q* or equivalent, or deionized
water
Site sediment(s), a reference sediment, and control
sediment
N/A
Bioaccumulation
<6 wk
1 L, minimum
Adequate mass of organisms at test completion for
detection of target analyte(s)

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                                              E-49

REFERENCES:

Lee II, H., B. Boese, J. Pelletier, M. Winsor, D. Specht and R. Randall. 1989. Guidance Manual: Bedded
    Sediment Bioaccumulation Tests. EPA/600/fc-89/302. 232 pp. (ATS Deliverable).

Bender, K. and W.R. Davis. 1984. Effects of feeding on Yoldia limatula on bioturbation. Ophelia 23: 91-
    100.

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                                            E-50
     SUMMARY OF TEST CONDITIONS AND TEST ACCEPTABILITY CRITERIA FOR THE
              AMPHIPOD, Diporeia sp., SEDIMENT BIOACCUMULATION TESTS
1.   Test type:
2.   Test duration:
3.   Temperature:
4.   Salinity:
5.   Light quality:
6.   Light intensity:
7.   Photoperiod:
8.   Test chamber size:
9.   Test solution volume:
10.  Sediment depth:
11  Renewal of test solutions:
12.  Age of test organisms:
13.  No. organisms per test chamber:
14.  No. replicate chambers per sediment:
IS.  No. organisms per sediment:
16.  Feeding regime:
17.  Test chamber cleaning:
18.  Test solution aeration:

19.  Dilution water:

20.  Test concentrations:

21.  Dilution series:
22.  Endpoint:
23.  Sampling and sample holding requirements:
24.  Sample volume required:
25.  Test acceptability criterion:
Static Non-renewal or Overlying Water Renewal
28d
4°C
0-20 %o
Red darkroom light
Low
Continuous
4 L minimum
to4L
3cm
Variable
Mixed age juveniles
5 g (-500-1000) (minimum)
4 minimum
N/A (>10g OC/g organism)
None
None
If needed to maintain DO> 40% saturation
(<  100 bubbles/min.)
Moderately hard water; synthetic water modified
to reflect receiving water hardness or salinity to
20%
Site sediment, a reference sediment and a control
sediment
N/A
Bioaccumulation
<6wk
8L
Adequate mass of organisms at test completion for
detection of target analyte(s)

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                                              E-51

REFERENCES:

Landrum, P.F. 1989. Bioavailability and toxicokinetics of polycyclic aromatic hydrocarbons sorbed to
    sediments for the amphipod, Pontoporeia hoyi.  Environ. Sci. Technol. 23:588-595.

Landrum, P.F., BJ. Eadie and W.R. Faust. 1991. Toxicokinetics and toxicity of a mixture of sediment-
    associated polycyclic aromatic hydrocarbons to the amphipod Diporeia spp.  Environ. Toxicol. Chem.
    10:35-46.

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       APPENDIX F
   METHODOLOGIES FOR
IDENTIFYING AMMONIA AS A
  TOXICANT IN DREDGED-
 MATERIAL TOXICITY TESTS
                               DRAFT

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DRAFT

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	F-l	

                                           APPENDIX F

                                Ammonia Toxicitv: General Overview

Ammonia is a relatively toxic compound which, in sediments, is generated from the microbial degradation of
nitrogenous organic material such as amino acids (Santschi et al, 1990).  Resulting interstitial (pore) water
concentrations of ammonia in otherwise uncontaminated sediments can be as high as 50 mg/L (Murray et al.,
1978; Kristensen and Blackburn, 1987), while ammonia concentrations in pore water from contaminated
sediments may range from SO to greater than 200 mg/L (Ankley et al.,  1990; Schubauer-Berigan and Ankley,
1991). Hence, exposure of epibenthic/benthic test species to ammonia in solid phase tests can be significant.
Moreover, because ammonia is released from sediments relatively readily  during resuspension events (Blom
et al., 1976), high concentrations can also occur in test elutriates. Both  marine and freshwater studies suggest
that ammonia can be responsible for toxicity observed in some laboratory sediment toxicity tests (Jones and
Lee, 1988; Ankley et al., 1990).

Because ammonia is not extremely persistent, its toxicity may not be  of as much concern as that from, for
instance, metals or pesticides. For this reason, there has been a tendency in some situations to use open-water
disposal for dredged material whose toxicity is suspected to be due to ammonia. Unfortunately it has previously
been  difficult, if not impossible, to validly  link sediment or elutriate toxicity to ammonia when multiple
sediment contaminants are present (Ankley et al., 1992), in particular  because ammonia concentrations can
be exceptionally high in sediments which are also toxic due to other, persistent contaminants such as inorganic
and/or organic chemicals (Schubauer-Berigan and Ankley, 1991). However, recent technical developments have
resulted in a logical conceptual framework, specifically a simple risk assessment, for deciding whether observed
sediment (or elutriate) toxicity may be due to ammonia. Briefly, data are collected on the toxicity of ammonia
to the test species of concern (effects assessment), and concentrations of ammonia are measured in appropriate
test fractions (elutriate, overlying  water, pore water)  during the  toxicity test  (exposure assessment). If
concentrations of ammonia in the test are large enough to result in toxicity to the test species of concern (risk
characterization), a simple set of toxicity identification evaluation (TIE) procedures is next used to confirm
that toxicity is indeed due to ammonia and not to other contaminants  in the sediment (Ankley et al., 1992).
TIE methods consist of physical/chemical sample manipulations conducted concurrently with toxicity testing
in order to directly characterize and identify contaminants responsible for toxicity in complex mixtures. Further
information on how this  approach could be used,  and important technical considerations relative to this
assessment are described below.
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	F-2	

              Specific Considerations for Assessing Ammonia Toxicltv In Dredged Material

The first step in assessing  the potential  for ammonia toxicity in a sediment  test is to routinely measure
ammonia concentrations in test fractions of concern, at a minimum, when starting and ending the test. Due
to the influence of pH on ammonia toxicity to some species, it is essential that pH also be measured and
recorded simultaneously. For elutriate tests, ammonia measurements can be made on whole elutriate. For solid
phase tests, ammonia should be measured both in overlying water and in pore water, the potential routes of
ammonia exposure for epibenthic and benthic species. In tests where periodic renewal of overlying water is
utilized, ammonia may not be present at lexicologically significant concentrations in the overlying water
(Ankley et al., 1993); nonetheless, it would still be prudent to measure ammonia in the overlying water.
Regardless of whether overlying water renewal is used in a sediment test, pore water ammonia concentrations
should be determined. Pore water for ammonia measurements can be isolated using any of a variety of
techniques (e.g., low-speed centrifugation, squeezing, peepers, etc.). Unlike other pore water contaminants of
concern (e.g., metals, nonionic organics), it does not appear that the method used to isolate pore water greatly
affects observed ammonia concentrations (EPA, 1991a). Upon isolation of the appropriate test fraction(s),
ammonia can be measured using any accepted technique; specific ion electrodes are rapid, simple and often
used for ammonia determinations at concentrations ;> 1 mg/L (EPA, 1979).

The next step is to compare exposure data (i.e., ammonia measurements) to toxicity data. The basis of this
comparison most generally will be to ammonia  toxicity data generated in water-only toxicity tests.  For the
elutriate tests the comparison can be made directly while, for solid phase tests, the water-only toxicity data are
compared to overlying water and/or pore water ammonia concentrations. To assess the potential for ammonia
toxicity in a test with a given species, it is essential that comparisons be made to toxicity data generated with
that same species in tests conducted under conditions reasonably similar to the sediment test. The tendency
to attempt to extrapolate toxicity  data for one species to another species should be avoided. Such an approach
may be appropriate for some types  of risk analyses; however, for the  approach described here, this type of
extrapolation likely would result  in erroneous conclusions. Similarly, comparisons within a species should be
made only between tests which were conducted  under a relatively similar set of conditions. For example, it
would be inappropriate to compare toxicity data and ammonia concentrations from a short-term sediment test
to water-only chronic toxicity data for that same species. In addition to test length, pH is of primary concern
while hardness, salinity and temperature are of somewhat lesser concern. All of these factors can markedly
influence ammonia toxicity, and  must be accounted for to enable among test comparability.

Although there is a good deal of data on the toxicity of ammonia to various aquatic species (EPA, 1985), much
of this information was generated using pelagic species (e.g., cladocerans, fishes), which precludes comparison
to sediment exposures with commonly tested benthic species (e.g., amphipods). [Although  it should be noted
that these data would be useful for extrapolation to elutriate tests which commonly utilize pelagic species].
                                                                                    DRAFT

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	F-3	

Water-only toxicity data are available for some epibenthic/benthic species of concern, however these water-only
tests were often conducted under conditions quite different from those commonly used in sediment tests, which
greatly limits any extrapolation to sediment tests. However, efforts are now underway to generate useful data.
For example, toxicity data now exist for ammonia at four different pHs (ca., 6.5, 7.2, 7.8, 8.6) for Hyalella
azteca, Chironomus tentans and Lumbriculus variegatus (EPA, 1991a; G. Ankley, unpublished data). Ammonia
toxicity data have also been developed for the commonly tested marine amphipods Rhepoxynius abronius,
Eohaustorius estuarius,Ampelisca abdita and Grandidierella japonica in four-day water-only exposures at a pH
of 8.0 (Kohn  et al., 1993); and, ammonia toxicity data have been generated  for  the  polychaete Nereis
(Neanthes) arenaceodentata (Dillon et al., 1993).

Although toxicity data exist for several pelagic and some benthic species of concern, it may be necessary for
laboratories conducting dredged material tests with a particular species, under a given set of test conditions,
to develop ammonia toxicity data relevant to their species/test conditions. This  likely would be  a  wise
investment of resources, in particular for those laboratories conducting large numbers of tests with dredged
material.

In this regard, a major caution must be noted concerning pH in ammonia tests. Ammonia acts as a basic
compound in water. The un-ionized form (NH3) predominates at pH values greater than 9.3, while the ionized
form (NH<+) is most abundant at pH values less than 9.3. Through the pH range of 6 to 8.3 (which is typically
encountered in  freshwater and marine sediment tests), the percentage of un-ionized ammonia changes
approximately 250-fold. Based on models developed primarily with fish, it has been common to express
ammonia toxicity data  on an un-ionized (i.e., NH3) rather than a  total  (i.e.,  NH3  plus NH/) basis.  This
implicitly suggests that ionized ammonia is not of great lexicological significance. While this appears to be true
for fish (EPA, 1985), it does not appear to be the case for some invertebrates. For  example, H. azteca displays
the same sensitivity to total ammonia (NH3 plus NH4+) over a pH range of approximately 6.0 to 8.5, suggesting
that this amphipod is very sensitive to ammonium ion (EPA, 1991a; G. Ankely,  unpublished data). Hence,
extrapolation of ammonia  toxicity data collected at only one pH  value, based on un-ionized ammonia
concentrations, would  result in  inaccurate predictions of potential toxicity of  ammonia to at least this
amphipod. Other invertebrates may exhibit a similar lack of predictability relative to pH/ammonia interactions.
Unfortunately, relatively few ammonia toxicity tests with invertebrates have been conducted at multiple pHs;
thus,  it is difficult to broadly predict responses to ammonia at different pH values.  To make accurate
predictions of potential ammonia toxicity for a particular test species, it is important to obtain  (or generate)
ammonia toxicity data within the pH range in which extrapolations are made.

If measurements of ammonia in elutriate tests,  or overlying and/or pore water in solid phase tests  are
determined to be of possible lexicological significance, it is essential that the role of ammonia in causing
toxicity be confirmed. It is importanl to avoid the tendency to assume that if a dredged material test exhibits
                                                                                    DRAFT

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	F-4	

toxicity and ammonia is present, that ammonia is the sole (or even major) cause of the observed toxicity. Toxic
concentrations of other  contaminants may be present simultaneously with ammonia.  In  such cases  the
assumption that only ammonia was causing toxicity could lead to disposal decisions (i.e., open-water) that may
result in serious long-term impacts to benthic communities.

Relatively simple TIE manipulations as generally described by Ankley et al. (1992), and specifically in a series
of guidance manuals (EPA,  1988; 1989a; 1989b; 1991a; 1991b) may be used to determine whether (or not)
ammonia is responsible for the observed toxicity. To date, these TIE methods have only been used with
freshwater sediments. However, in many instances similar approaches can be used with marine sediments; also,
EPA currently is developing standardized TIE methods for marine sediments.

Current sediment TIE methods are only for elutriates or pore waters  and for short-term (s 96  hour) tests
(EPA, 1991a). This is not a problem if TIE procedures are to be used with toxic elutriates, because elutriate
tests also generally consist only of short-term (* 96 hour) exposures. However, solid phase tests with dredged
material are generally 10 days in length. Although using pore water as a surrogate test fraction for TIE work
with solid phase exposures could mean that toxicity might not be expressed in the shorter-term  pore water
exposures, this may not be a significant problem in the case of ammonia. In water-only exposures with three
different benthic invertebrates (H. azteca, C. tertians, L. variegatus), the majority of toxicity due to ammonia
was observed within 4 days in a 10 day test (G. Ankley, unpublished data).

There is one other important consideration relative to the use of pore water as a surrogate test fraction for
solid phase  sediments. Because the toxicity of ammonia to  some organisms  can  be pH-dependent,  it is
imperative that pH in pore water tests mimic the pH in the initial solid phase tests. This is particularly
important with freshwater sediments, because pH can drift upwards by as much as one unit over the course
of a 96-hour  test (Ankley et al.,  1991). Methods which have proven useful for controlling pH in  pore water
tests include: (a) use of acids/bases in chambers with minimal head-space, (b) use of organic  buffers, and (c)
use of varying amounts of CO2 in head-space overlying the pore water  (EPA, 1991a; 1991b).

Most aquatic species that can be tested successfully in a water-only exposure can be utilized for TIE work. For
example, cladocerans (Ceriodaphnia dubia, Daphnia magna, Daphnia pulex), fish (Pimephalespromelas, Oryzias
latipes, Oncorhynchus mykiss), amphipods (H. azteca), oligochaetes (L. variegatus), and chironomids (C. tentans)
all have been used for freshwater TIE studies. The best  choice of a TIE organism is, of course, the same
species that was sensitive to  the original elutriate or solid phase sediment of interest. For example, if toxicity
was observed in solid phase sediment tests with H. azteca, that species would be the best choice for pore water
TIE work. Of course, there are instances in which this may not be possible; for example, the test species of
concern may be of limited availability. In this case, it may be  possible  to use surrogate species for the TIE,
provided there is adequate knowledge of the sensitivity of the surrogate species to ammonia, relative to the
                                                                                     DRAFT

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	F-5	

original test species of concern. If a surrogate species is used, upon conclusion of the TIE it is important to
perform limited testing to confirm that the same compound(s) which was toxic to that species was responsible
for toxicity to the original test species.

As discussed above, the toxicity of ammonia to many species can be highly pH-dependent. If the test species
of concern is more sensitive to un-ionized that ionized ammonia, samples will be more toxic at high pH values
than at low pHs. [Note that this again demonstrates the need for data concerning pH/ammonia interactions
for specific test organisms]. If the test species exhibits this pH-dependency with regard to ammonia toxicity,
the graduated pH test can be  an extremely powerful tool for implicating ammonia as a suspect toxicant. The
graduated pH test is conducted at a series of physiologically tolerable pHs (generally ranging from 6.0 to 8.5);
if sample toxicity is greater at higher pH values, this suggests that ammonia is responsible  for at least some
of the observed toxicity. A number of other TIE techniques also exist for implicating ammonia. These include
evaluation of relative species sensitivity (e.g., fish are generally more sensitive than cladocerans), removal of
ammonia  from the test samples with cation  exchange resins  (e.g., zeolite)  and/or extended air-stripping at
elevated pH values (e.g.,  >10) prior to toxicity testing, correlation of toxicity with  measured ammonia
concentrations and toxicity tests at different pH values with equitoxic concentrations of ammonia (EPA, 1989a;
Ankley et al.,  1990).  Another  useful method for  confirming that ammonia is responsible for toxicity is
ammonia removal followed by spiking to restore the original ambient concentrations of ammonia. The spiked
sample is  then tested for toxicity; if ammonia is the causative  toxicant, observed toxicity theoretically should
be the same as that observed in the original sample. It is desirable to conduct as many of these confirmation
tests as possible because no single test is specific for ammonia,  e.g., zeolite will remove cationic metals, as well
as ammonia, from test samples. Failure of one or more of the tests to confirm ammonia as responsible for
toxicity would  indicate that other contaminants were contributing to sample toxicity.

                                             Summary

In order to identify elutriate or solid phase dredged material toxicity due to ammonia, it is essential  to make
routine measurements of ammonia on appropriate test fractions. These measurements then are compared to
water-only toxicity data for the same species  used in the dredged material  test. The water-only toxicity data
should be generated under conditions (e.g., pH, test length) reasonably similar to those in  the test with the
dredged material. If ammonia concentrations are too low to have potentially caused the observed toxicity in
the dredged material sample, other contaminants are responsible for the toxicity. If ammonia concentrations
are high enough to have caused the observed toxicity, TIE procedures should be used to confirm this suspicion.
When there is no TIE confirmation that ammonia is  responsible for sediment toxicity, it must be assumed that
persistent  contaminants other than ammonia are causing toxicity.
                                                                                    DRAFT

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	F-6	

                                         References Cited

Ankley, G.T., A. Katko and J.W. Arthur. 1990. Identification of ammonia as an important sediment-associated
        toxicant in the lower Fox River and Green Bay, Wisconsin. Environ. Toxicol. Chem. 9:313-322.

Ankley, G.T., M.K. Schubauer-Berigan and J. R. Dierkes. 1991. Predicting the toxicity of bulk sediments to
        aquatic organisms with aqueous test fractions: pore water versus elutriate. Environ. Toxicol. Chem.
        10:1359-1366.

Ankley, G.T., M.K.  Schubauer-Berigan  and R.A. Hoke. 1992.  Use  of toxicity identification evaluation
        techniques to identify dredged material disposal options: A proposed approach. Environ. Manage.
        16:1-6.

Ankley, G.T., D.A.  Benoit, R.A. Hoke, E.N. Leonard, C.W. West, G.L. Phipps, V.R. Mattson and L.A.
        Anderson. 1993. Development and evaluation of test methods for benthic invertebrates and sediments:
        Effects of flow rate and feeding on water quality and exposure conditions. Arch. Environ. Contain.
        Toxicol. (In Press).

Blom,  B.E., T.F.  Jenkins, D.C. Leggett and R.P. Murrmann. 1976. Effect of sediment organic matter on
        migration of various chemical constituents during disposal of dredged material. Report No. D-76-7.
        U.S. Army Corps of Engineers Waterways Experiment Station, Vicksburg, MS.

Dillon, T.M., D.W. Moore and AB. Gibson. 1993. Development of a chronic sublethal sediment bioassay with
        the marine polychaete Nereis (Neanthes) arenaceodentata. Environ. Toxicol. Chem. 12:589-605.

EPA.  1979. Methods for Chemical Analysis of Water and Wastes.  EPA/600/4-79-020. U.S. Environmental
        Protection Agency, Cincinnati, OH.

EPA.  1985. Ambient Water Quality Criteria for Ammonia -  1984. EPA  440/5-85-001. U.S. Environmental
        Protection Agency, Office of Water Regulations and Standards, Washington, DC.

EPA. 1988. Methods for Aquatic Toxicity Identification Evaluations: Phase I Toxicity Characterization Procedures.
        EPA/600/3-88-034.  U.S. Environmental Protection Agency, Environmental Research Laboratory,
        Duluth, MN.
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	F-7	

EPA. 1989a. Methods for Aquatic Taxicity Identification Evaluations: Phase II Toxicity Identification Procedures.
       EPA/600/3-88-035. U.S. Environmental  Protection Agency, Environmental Research Laboratory
       Duluth, MN.

EPA 1989b. Methods for Aquatic Toxicity Identification Evaluations: Phase III Toxicity Confirmation Procedures.
       EPA/600/3-88-036. U.S. Environmental  Protection Agency, Environmental Research  Laboratory,
       Duluth, MN.

EPA. 1991a. Sediment Toxicity Identification Evaluation: Phase I (Characterization), Phase II (Identification) and
       Phase III (Confirmation) Modifications of Effluent Procedures. EPA/600/6-91-007. U.S. Environmental
       Protection Agency, Environmental Research Laboratory, Duluth, MN.

EPA 1991b. Methods  for Aquatic  Toxicity  Identification Evaluations: Phase  I  Toxicity  Characterization
       Procedures.  2nd Ed. EPA/600/6-91/003. U.S. Environmental  Protection Agency,  Environmental
       Research Laboratory, Duluth, MN.

Jones, R.A and  G.F. Lee. 1988. Toxicity of U.S. waterway sediments with particular reference to the New
       York Harbor area. Pp.  403-417. In: J.J. Lichtenberg, F.A Winter, C.I. Weber and L. Fradkin (Eds).
       Chemical and Biological Characterization of Sludges, Sediments, Dredged Soils and Drilling Muds. ASTM
       STP 976. American Society for Testing and Materials, Philadelphia, PA

Kohn, N.P., J.Q.  Ward,  D.K. Nigogi, L.T. Ross, T. Dillon and D.W. Moore. 1993. Acute toxicity of ammonia
       to four species  of marine amphipod. Submitted Manuscript.

Kristensen, E. and T.H. Blackburn. 1987. The fate of organic carbon and nitrogen in  experimental marine
       sediment systems: Influence of bioturbation  and anoxia. J. Mar. Res. 45:231-257.

Murray, J.W., V. Grundmanis and W.M. Smethie, Jr. 1978.  Interstitial water chemistry in the sediments of
       Saanich Inlet. Geochim. Cosmochim. Acta. 42:1011-1026.

Santschi, P., P. Hohener, G. Benoit and M.B. Brink. 1990. Chemical processes at the sediment-water interface.
       Mar.  Chem. 30:269-315.

Schubauer-Berigan, M.K. and G.T. Ankley. 1991. The contribution of ammonia, metals and nonpolar organic
       compounds to  the toxicity of sediment interstitial water from  an Illinois river tributary. Environ.
       Toricol.  Chem. 10:925-939.
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F-8
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      APPENDIX G
  QUALITY ASSURANCE/
QUALITY CONTROL (QA/QC)
    CONSIDERATIONS
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                             TABLE OF CONTENTS
                                                                              Page No.

Table of Contents                                                                       i
List of Tables                                                                         ii
List of Figures                                                                         ii

G.O          QUALITY   ASSURANCE/QUALITY   CONTROL   (QA/QC)
             CONSIDERATIONS  	G-l
G.I          Introduction	G-l
G.I.I        Government (Data User) Program  	G-2
G.1.2        Contractor (Data Generator) Program	G-2
G.2          The QA Project Plan	G-3
G.2.1        Project Description  	G-4
G.2.2        QA Organization; Personnel Responsibilities and Qualifications	G-4
G.2.3        Data Quality Objectives	G-5
G.2.4        Standard Operating Procedures	G-5
G.2.5        Sampling Strategy and Procedures  	G-6
G.2.6        Sample Custody and Documentation	G-6
G.2.6.1             Field Operations	G-7
G.2.6.2             Laboratory  Operations  	G-7
G.2.7        Calibration Procedures	G-8
G.2.8        Analytical Procedures	  G-8
G.2.9        Data Validation, Reduction and Reporting	G-8
G.2.10       Internal Quality Control Checks	G-9
G.2.10.1           Quality Control Considerations for Physical Analysis of Sediments .  G-10
G.2.10.2           Quality  Control   Considerations   for   Chemical  Analysis   of
                   Sediments   	  G-ll
G.2.10.3           Quality Control Considerations for Chemical Analysis of Water ...  G-ll
G.2.10.4           Quality Control Considerations for Chemical Analysis of Tissue . . .  G-12
G.2.10.5           Quality Control Considerations for Biological Analyses  	  G-12
G.2.10.5.1                 Source and Condition of Test Organisms	  G-12
G.2.10.5.2                 Reference Toxicants	  G-13
G.2.10.5.3                 Acceptability of Test Results and Data Evaluation	  G-1S
G.2.11       Performance and System Audits	  G-15
G.2.11.1           Pre-award Inspections  	  G-16
G.2.11.2           Interlaboratory Comparisons (Chemical Analytical Laboratories) . .  G-16
G.2.11.3           Routine Inspections	  G-19
G.2.12       Facilities	  G-19
G.2.13       Preventive Maintenance	  G-19
G.2.14       Calculation of Data Quality Indicators  	  G-20
G.2.15       Corrective Actions  (Management of Nonconformance Events)  	  G-20
G.2.16       QA Reports to Management  	  G-21
G.3          REFERENCES	  G-21
G.4          EXAMPLE QA/QC CHECKLISTS, FORMS AND RECORDS	  G-23
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                                      11
                             LIST OF TABLES
                            LIST OF FIGURES
                                                                     Page No.



Table G. 1     Sources of Standard Reference Materials                               G-18
Figure G.I    Example Control Charts for Reference Toxicants.                         G-14
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	G-l	

G.O          QUALITY ASSURANCE/QUALITY CONTROL (QA/QC) CONSIDERATIONS

G.I          Introduction

The following sections provide interim guidance for QA/QC. They will be replaced, in due course, by
a separate QA/QC document applicable to both the present manual and the Ocean Disposal "Green Book"
(EPA/USACE, 1991). Phase 1  of the QA/QC guidance, pertaining to chemical evaluations,  will be
available in 1994 (EPA, 1994). It will: 1) provide guidance on the development of QA project plans for
ensuring the reliability of data gathered to evaluate dredged material proposed for discharge under the
CWA or the MPRSA; 2) outline procedures that need  to be followed when sampling and analyzing
sediments, water,  and tissues; and 3) provide recommended target detection limits (TDLs) for chemicals
of concern.  Phase 2 of the QA/QC  guidance,  pertaining to biological evaluations,  will  follow the
publication of Phase 1.

A quality assurance (QA) program integrates management and technical practices into a single system to
guarantee quality environmental data.  The purpose of a QA program in a dredged material evaluation is
to provide environmental  data that are sufficient, appropriate, and of known and documented quality.
Major elements of a QA program are:

       •     human resource training
       •     QA management plan (QAMP)/QA project plan (QAPP)
       •     management system reviews
       •     data quality objectives (DQOs)
       •     standard operating procedures (SOPs)
       •     project specific technical assessments.

QA project plans provide, in one place, a detailed plan for the activities performed at each stage of the
dredged material evaluation (including appropriate sampling and analysis procedures) and outline project-
specific data quality objectives that should be achieved for field observations and measurements, physical
analyses, laboratory chemical analyses, and biological tests. Data quality objectives must be defined prior
to initiating a project and  adhered to for the duration of the project in order to guarantee acquisition of
reliable data. This is accomplished by integrating  quality control (QC) into all facets of the project,
including development, implementation, and evaluation. QC is the routine application of procedures for
determining bias and precision. QC procedures include activities such as preparation of replicate samples,
spiked  samples, blanks;  calibration and standardization; sample custody  and recordkeeping.  Audits,
reviews and compilation of complete and thorough documentation are activities used to verify compliance
with pre-defined  QC procedures. Through periodic reporting, these activities  provide a means for
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	G-2	

management to track project progress and milestones, performance of measurement systems, and data
quality.

A complete QA/QC effort for a dredged material testing program has two major components:  a QA
program implemented  by the responsible governmental agency (the data user),  and QC programs
implemented  by sampling and laboratory  personnel performing the tests (the data generators).  QA
programs  are also implemented by each field contractor and each laboratory. Typically, all field and
laboratory data generators agree to adhere to the QA/QC of the data user for the contracted project as
specified in the project QAPP. EPA (1987) provides useful guidance and may be followed on all points
that are not in conflict with the guidance in this manual.
G.I.I          Government (Data User) Program

The USAGE must implement a QA program to ensure that all program elements and testing activities
(including field and laboratory operations) in the dredged material evaluation comply with the procedures
in the QA project plan or with other specified guidelines for the  production of environmental data of
known quality. QA oversight is the responsibility of the USAGE District Office, working in conjunction
with the EPA  Region. USAGE Districts are responsible for ensuring that both the data submitted with
permit applications, and that laboratories under contract to their Districts comply with the QA needs of
the regulations and guidelines governing dredged material evaluations. The QA program should be
designed with the assistance of programmatic and scientific expertise from both EPA and USAGE. Other
qualified sources of QA program management should be contacted as  appropriate. Some specific QA
considerations in contract laboratory selection are discussed by Sturgis (1990) and EPA (1991a).
G.I.2          Contractor (Data Generator) Program

Each  office or laboratory participating in a  dredged material  evaluation is  responsible for using
procedures which assure that the accuracy  (precision and bias), representativeness, comparability, and
completeness of its data are known and documented.  To ensure that this responsibility  is met, each
participating organization  should have a project manager and a written QA management  plan that
describes, in specific terms, the management approach proposed to assure that each procedure under its
direction complies with the criteria accepted by EPA and USAGE. This plan  should describe a QA
policy,  address the contents  and application  of  specific  QA  project  plans,  and specify  training
requirements.  All field  measurements,  sampling, and  analytical components  (physical, chemical, and
biological) of the dredged  material evaluation should be discussed.
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	G-3	

For the completion of a dredged  material testing project, the project manager of each participating
organization should establish a well-structured QA program that ensures the following:

       •       development, implementation, and administration of appropriate QA planning documents
               for each study

       •       inclusion of routine QC procedures for assessing data quality in all field and laboratory
               standard operating procedures (SOPs)

       •       performance  of  sufficiently detailed  audits at intervals frequent enough to ensure
               conformance with approved QA project plans and SOPs

       •       periodic evaluation of QC procedures to improve the quality of QA project plans and
               SOPs

       •       implementation of appropriate corrective actions in a timely manner.
G.2           The QA Project Plan

The QA project plan should be developed by the applicant or  contractor for  each  dredged material
evaluation, in accordance with EPA (1994). The QA project plan provides an overall  plan and contains
specific guidelines and  procedures for the activities performed at each  stage of the  dredged material
testing program, such as dredging site subdivision, sample collection, bioassessment procedures, chemical
and physical analyses, data quality standards, data analysis and  reporting. In particular, the QA plan
addresses required QC checks, performance and system audits, QA reports to management,  corrective
actions, and assessment of data accuracy (precision and bias), representativeness, comparability and
completeness. The plan should address the quantity of data required to allow confident and justifiable
conclusions and decisions.  QA project plans are particularly useful for work that involves many people
or for projects that continue over a long period. When many people are involved, the plan ensures that
everyone  has a thorough understanding of the goals and procedures  of the program.  When work is
conducted over a long period, the plan provides a basis for continuity, ensuring that procedures do not
slowly change over time without the persons involved in the program evaluating the nature of the changes
and their possible impact on data quality.

Each of the following items should be considered  for inclusion in the QA Project Plan:

       •       Project description (G.2.1)
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              	G-4	

               QA organization; personnel responsibilities and qualifications (G.2.2)
               QA  objectives for  measurement data  in  terms  of  accuracy,  representativeness,
               comparability, and completeness (G.2.3)
               Standard operating procedures (G.2.4)
               Sampling strategy and procedures (G.2.S)
               Sample custody and documentation (G.2.6)
               Calibration procedures (G.2.7)
               Analytical procedures (G.2.8)
               Data validation, reduction and reporting (G.2.9)
               Internal  QC checks (G.2.10)
               Performance and system audits (G.2.11)
               Facilities (G.2.12)
               Preventative maintenance (G.2.13)
               Calculation of data quality indicators (G.2.14)
               Corrective actions (G.2.15)
               QA reports to management (G.2.16).
G.2.1          Project Description

A project description should be provided that defines project goals and illustrates how the project will
be designed to  obtain the information needed to achieve those goals. Sufficient detail and information
should be included to allow decisions during the joint EPA and USAGE review and the final USAGE
approval phases. Where appropriate,  the following information should be included in this section of the
QA project plan:

        •      objectives and scope  of the project
        •      any historical information relevant to the dredging operation
        •      intended  activities further described  in flow diagrams, tables, and charts
        •      schedule of tasks and milestones
        •      intended  use of acquired data.
G.2.2         QA Organization; Personnel Responsibilities and Qualifications

A clear delineation  of the QA organization and line of authority is essential  for the development,
implementation, and administration of a QA program.  This should include  all technical personnel,
including key individuals responsible for ensuring sufficient QC is being incorporated into the project.
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	G-5	

Organizational  charts or tables should be used  in the QA project plan to describe the management
structure, personnel responsibilities, and the interaction among functional units. Each QA task should be
fully described and the responsible individual and associated organization named. An example of a QA
organization flow diagram is provided in Appendix G.4.

Technical staff are responsible for the validity and integrity of the data produced. The QA staff should
be responsible for ensuring that all personnel performing tasks related to data quality are appropriately
qualified. Records of qualifications and training of personnel should be kept current for verification by
internal QA personnel or by EPA and USAGE.
G.2.3          Data Quality Objectives

Data quality objectives are used to ensure that the data are acceptable. They define performance-based
goals for accuracy (precision and  bias), representativeness, comparability, and completeness as well as
the required sensitivity of chemical  measurements  (i.e., target detection limits, TDLs). Accuracy  is
defined in terms of bias (how close the measured value is to the true value) and precision (how variable
the measurements are when repeated). Data quality objectives should be based on the intended use of the
data, technical feasibility, and consideration of cost. Numerical quality objectives should be summarized
in a table,  with all data calculated and reported in units consistent with other organizations reporting
similar data, to allow comparability of data bases. All measurements should be made so that results are
representative of the medium (e.g., water, sediments, tissue) being measured. Data quality objectives for
precision and bias established for each measurement parameter should be based on prior knowledge of
the measurement system employed,  method validation  studies,  and  the requirements of the  specific
project. An example of a data quality objectives summary for laboratory measurements is provided in
Appendix G.4.
G.2.4         Standard Operating Procedures

Standard operating procedures (SOPs) are written descriptions of routine methods and should be provided
for as many methods used during the dredged material evaluation as possible. A large number of field
and  laboratory operations  can  be standardized and presented as SOPs.  Once  these procedures are
specified, they can be referenced or provided in an appendix of the QA project plan. Only modifications
to SOPs or non-standard procedures need to be explained in the main body of the QA project plan (e.g.,
in the "sampling procedures" or "analytical procedures" section). General types of procedures benefiting
from SOPs are field measurements ancillary to  sample  collection (e.g.,  depth of overlying water,
sampling  depth, water  quality  measurements, mixing model  input measurements),  chain-of-custody,
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	G-6	

sample handling and shipment, and routine analytical methods for chemical analyses. SOPs ensure that
all persons conducting work are following the same procedures and that the procedures do not change
over time. All personnel should be thoroughly familiar with the SOPs before work is initiated. Deviations
from SOPs may affect data quality and integrity. If it is necessary to deviate from approved SOPs, these
deviations must be documented and approved through an appropriate chain-of-command which may
include USAGE and EPA. Personnel responsible for ensuring the SOPs are adhered to must be identified
in the QA Project Plan. Example SOPs are provided in Appendix D of EPA (1994).
G.2.5         Sampling Strategy and Procedures

A sampling strategy should be developed to ensure that the sampling design supports the planned data
use. The sampling strategy will strongly affect the representativeness, comparability, and completeness
that might be expected for field measurements. In addition, the strategy for collecting field QC samples
(e.g., replicates) will assist in the determination of how well the total variability of a field measurement
can be documented. Therefore, development of the sampling strategy should be closely coordinated with
development of data quality objectives discussed in Section G.2.3.

To reduce  sampling error, all methods, procedures, and equipment to be used in the field should be
documented in a sampling plan which has been authorized and which is readily available to all personnel.
The purpose of this sampling plan is to provide a blueprint for all field work by defining in detail the
appropriate sampling  and data collection methods (in accordance with the established data quality
objectives). Written procedures or checklists for field equipment, sample container preparation, sample
preservation, labelling and numbering systems, and shipping procedures must be appropriate. Methods
to record and report deviations from the sampling plan must also be described. An alteration checklist
form is generally appropriate to implement required changes. An example of such a checklist is provided
in Appendix G.4.
G.2.6         Sample Custody and Documentation

Sample custody and documentation are vital components of all dredged material evaluations, particularly
if any of the data may be used in a court of law. It is important to record all events associated with a
sample so that the validity of the resulting data may be properly interpreted. Documentation is necessary
during the field effort when samples  are collected  and  in the laboratory where both chemical and
biological analyses are performed. Thorough documentation provides a means to track samples from the
field through the laboratory and prevent sample loss. The contents and location of all documents related
to dredged sediment samples should be specified, and  access to the samples should be controlled. Where
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	G-7	

samples may be needed for potential litigation, chain-of-custody procedures should be followed. Chain-of-
custody procedures are initiated during sample collection. They include a descriptive label and tracking
report forms for both the field and laboratory. An example of a label, field tracking report form,
laboratory tracking report form and chain-of-custody record is provided in Appendix G.4.
G.2.6.1               Field Operations

The potential for sample deterioration and/or contamination exists during sample collection, handling,
preservation, and storage. Approved protocols and SOPs should be followed to ensure all field equipment
is acceptably calibrated and to prevent deterioration or contamination. Experienced personnel should be
responsible for maintaining the sample integrity from collection through analysis. A complete record of
all field procedures, an inventory log, and a tracking log should be maintained. A field tracking report
should identify sample custody and conditions in the field prior to shipment.

Dates and times of collection, station locations, sampling methods, and sample handling, preservation,
and storage procedures should be documented immediately,  legibly, and indelibly so that they are easily
traceable. Any circumstances potentially affecting sampling  procedures should be documented. The data
recorded should be thorough enough to  allow  station relocation and sample tracking. An example of a
station location  log is provided in Appendix G.4. Any field preparation of samples should also be
described. Samples should be identified with a  pre-prepared label  containing at least the following
information:

       •      project title
       •      sample identification number
       •      location (station number) and depth
       •      analysis or test to be performed
       •      preservation and storage method
       •      date and time of collection
       •      special remarks if appropriate
       •      initials of person collecting the sample
       •      name of company performing the work.
G.2.6.2               Laboratory Operations

The responsible party who will act as sample custodian at the laboratory facility should be identified. This
individual has authority to sign for incoming field samples and has the responsibility to obtain documents
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	G-8	

of shipment and verify the data entered on the sample custody records.  A laboratory-tracking report
should be prepared for each sample. The location of samples processed through chain-of-custody must
be known at all times.  Samples to be used in a court of law must be stored  in a locked facility to prevent
tampering or alteration.

A procedure should be established for the retention  of all appropriate field and laboratory records and
samples as various tasks or phases are completed. Replicates, subsamples  of analyzed samples, or extra
unanalyzed samples should be kept in a storage bank. These samples can be used to scrutinize anomalous
results or for supplemental analyses, if additional information is needed. All samples should be properly
stored and inventoried. The retention and archiving procedure should indicate the storage requirements,
location, indexing codes, retention time, and security requirements for samples and data.
G.2.7         Calibration Procedures

Calibration procedures should be included for each instrument used during the study. The appropriate
procedures used to assure that field  and laboratory  equipment are functioning properly should be
documented in this section. This information can be provided in tabular format. The planned frequency
for recalibration should be provided as well  as a list of the calibrations standards to be used and their
sources, including traceability procedures. Instrumentation that requires routine calibration includes, for
example, navigation devices, analytical balances, and water quality meters.
G.2.8         Analytical Procedures

The methods cited in the analytical procedures section of a QA project plan are used to meet the data
quality objectives for a dredged material evaluation. (Section 9 of this Manual provides guidance on the
selection of physical and chemical analyses to aid in evaluating dredged material proposed for disposal,
and on the methods used to analyze these parameters.)  In all cases,  proven,  state-of-the-art methods
should be used. Sample analysis  procedures  are identified in this section of the QA project plan by
reference to established, standard methods. Any modifications to established, standard methods and any
specialized, nonstandard procedures should be described in detail in this section of the plan.
 G.2.9         Data Validation, Reduction and Reporting

 Data validation involves all procedures used to accept or reject data after collection and prior to use.
 These include screening, editing, verifying, and reviewing through external performance evaluation
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	G-9	

audits. Data validation procedures ensure that objectives for data precision and bias were met, that data
were generated  in accordance with the QA project plan  and SOPs,  and that data  are traceable and
defensible. All data should be reported with their associated analytical sensitivity, precision, and bias.
In addition, the level of quantification achieved by the laboratory should be compared to specific target
detection limits. The following information should be  included in the QA project plan:

        •       the principal criteria that will be used to validate data integrity during their collection and
               reporting

        •       the data reduction  scheme planned for  collected data including all  equations used to
               calculate the concentration or value of the measured parameter and reporting units

        •       the methods used to identify and treat  outliers and nondetectable data

        •       the data flow or reporting  scheme from  collection of raw data through storage of
               validated concentrations (a flowchart is usually necessary)

        •       key individuals who will handle the data in this  reporting scheme.

QC procedures designed to eliminate  errors during the mathematical and/or statistical reduction of data
should also be included in the QA project plan. Quality control in data processing  may include both
manual and automated review. Input  data should be checked and verified to confirm compatibility and
to flag "outliers" for confirmation. Computerized data plots can be routinely used as a tool for  rapid
identification of outliers that can then be verified using standard analytical procedures.

Data entries should be dated when entered, and signed or initialled by the person making the measurement
and  the person entering the data. Changes to entries should be  made so as not to obscure the original
entry. They should indicate the reason for the change, the person  making the change,  and the date of
change. In computer-driven data collection systems, the person  responsible for direct data input should
be identified at the time of input.

The  data and information collected  during the Tier I evaluation should be carefully reviewed as to their
relevancy,  completeness, and  quality. The data must be relevant to the overall objective of the project,
even though the objectives for these studies were different.
G.2.10               Internal Quality Control Checks
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	G-10	

The various control samples that will be used internally by the laboratory or sample collection team to
assess quality are described in this section of the QA project plan. For most environmental investigations,
10-30 percent of all samples may be analyzed specifically for purposes of quality control. In some special
cases (e.g., when the number of samples is small and the need to establish the validity of analytical data
is large), as many as SO percent of all samples are used for this purpose. These QC samples may be used
to check the bias and  precision of the  overall analytical system and to evaluate the  performances of
individual analytical instruments or the technicians that operate them. The most widely used QC samples
are summarized in EPA (1994) and are  as follows:

        •       blanks

        •       matrix  spike samples

        •       surrogate spike compounds

        •       check standards, including:
                      spiked method blanks
                      laboratory control  samples
                      reference  materials

        •       matrix  replicates (split in the laboratory from one field sample)

        •       field replicates (collected as separate field samples from one location).

The following sections discuss quality control procedures for sediment, water, and tissue analyses (see
EPA, 1994 for further detail), as  well as for biological analyses.

The USAGE District or management authority  for the program may require that certain samples be
submitted on a routine basis to government laboratories for analysis, and EPA or USAGE may participate
in some studies. These activities provide  an independent  quality  assurance check on activities  being
performed and on data being generated  and are discussed in Section G.2.11.
G.2.10.1             Quality Control Considerations for Physical Analysis of Sediments

The procedures used for the physical analysis of sediments must include a QG component. QG procedures
for grain-size analysis and total solids/specific gravity determinations are necessary to ensure that the data
meet  acceptable criteria for  precision and bias.  To measure precision,  triplicate analyses should be
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	G-ll	

performed for every 20 samples analyzed. TOG is a special case, where all samples should be analyzed
in triplicate. In addition, one procedural blank per 20 samples should be run, and the results reported for
TOG analysis. Standards used for TOC determinations must be verified by independent check standards
to confirm the bias of the results. QC limits should be agreed upon for each analytical procedure, and
should be consistent with the overall QA project plan.
G.2.10.2              Quality Control Considerations for Chemical Analysis of Sediments

Methods for the chemical analysis of contaminants of concern  in  sediments must include detailed
procedures and requirements which should be followed rigorously throughout the evaluation. General
procedures include the analysis of a procedural blank, a matrix duplicate, a matrix spike along with every
10 - 20 samples processed, and surrogate spike compounds (for organic analyses only). All analytical
instruments should be calibrated at least daily. All calibration data should be submitted to the laboratory
project  QA coordinator for review. The QA/QC program must document the ability  of the selected
methods to address the high salt content of sediments from marine and estuarine areas.

Analytical precision can be measured by analyzing one sample in duplicate or triplicate for every  10 -
20 samples analyzed. If duplicates are  analyzed, the relative percent difference should be reported.
However, if triplicates are analyzed, the percent relative standard deviation should be reported.
G.2.10.3              Quality Control Considerations for Chemical Analysis of Water

Methods recommended for the chemical analysis of contaminants of concern in water include detailed QC
procedures and requirements which should be followed closely throughout the evaluations. General
procedures should include the analysis of a procedural blank, a matrix duplicate, a matrix spike for every
10 - 20 samples  processed,  and surrogate  spike compounds  (for organic  analysis only).  Analytical
precision can be measured by analyzing one sample in triplicate or duplicate for every 10 - 20 samples
analyzed.  If duplicates are analyzed, the relative percent difference should be reported. However, if
triplicates are analyzed, the percent relative standard deviation should  be reported. Analytical bias can
be measured by analyzing standard reference materials (SRMs), a matrix containing a known amount of
a pure reagent. Recoveries of surrogate spikes and matrix spikes should be used to measure for precision
and bias; results from these analyses should be well documented. Special QC is required for ICP and
GC/MS analyses. Initial calibrations using three or five standards (varying concentrations) are required
before analyzing samples. Subsequent calibration checks should be performed for every 10-20 samples
analyzed.
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	G-12	

G.2.10.4             Quality Control Considerations for Chemical Analysis of Tissue

As with sediments and water, methods recommended for the chemical analysis of contaminants of concern
in tissues include detailed QC procedures and requirements which should be followed closely throughout
the evaluations. General procedures should include the analysis of a procedural blank, a matrix duplicate,
a matrix spike for every 10 - 20 samples processed, and surrogate spike compounds (for organic analyses
only). Analytical precision can be measured by analyzing one sample in triplicate or duplicate for every
10 - 20 samples analyzed. If duplicates are analyzed, the relative percent difference should be reported.
However, if triplicates are analyzed, the percent relative standard deviation should be reported. Analytical
bias can be measured with the appropriate SRMs. Precision and bias determinations should be performed
with the same frequency as the blanks and  matrix spikes.
G.2.10.5             Quality Control Considerations for Biological Analyses

Quality controls for tests of biological effects and bioaccumulation must address all activities that affect
the quality of the data (e.g., see EPA,  1991b). These activities include:

        •      source and condition of test organisms

        •      use of negative (non-toxic) and positive (reference toxicants) controls

        •      acceptability of test results and data evaluation.

Standard laboratory procedures must be followed in all testing including maintenance/measurement of
environmental (e.g., water) quality conditions and blind testing.


G.2.10.5.1                   Source and Condition of Test Organisms

Test organisms  should be positively identified to species by qualified experts. Test organisms should
appear healthy, behave normally, feed well, and have low mortality in cultures, during holding, and in
test controls. The quality of test organisms from outside sources as well as those maintained in-house
must be verified by conducting a reference toxicant test concurrently with the dredged material toxicity
tests. The  supplier should provide data with the shipment describing the history of the sensitivity of
organisms  from the same source culture, determined in monthly tests using suitable reference toxicants.
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	G-13	

G.2.10.5.2                    Reference Toxicants

Biological QC includes periodic reference toxicant tests with all stocks of organisms to be used in testing
to determine the relative health of the organisms. The application and benefits of reference toxicant tests
are discussed by Lee (1980). Detailed assistance in establishing a biological QC program can be provided
by scientists from EPA or USAGE.

Reference toxicants are routinely used to evaluate species sensitivity, laboratory performance and both
intra- and niter- laboratory precision. The following chemicals provide good endpoints for a variety of
species:  freshwater species - sodium chloride, copper sulfate, potassium chloride, cadmium chloride,
sodium dodecyl sulfate, diazinon; saltwater species - copper sulfate, cadmium chloride, sodium dodecyl
sulfate, diazinon. It is required that  a set of the above chemicals with difference modes of toxic action
be used as reference toxicants in establishing comparative sensitivity between recommended species listed
in Tables 11, 12 and  13 of the Manual and a species proposed as a substitute regional test species.

Reference toxicant tests should be performed routinely on all groups of organisms used  in  dredged
material  toxicity and bioaccumulation  studies in  order to determine their relative health and vigor. A
single reference toxicant  can be used  to assess this in routine testing. Many chemicals may  be used
satisfactorily as reference toxicants (e.g., Lee,  1980; Wang,  1987; EPA, 1990,  199Ic).  Reference
toxicant tests are performed in the absence of sediment and generally under static conditions. Water-only
reference toxicant tests with benthic species may require some  modification to "standard" test conditions
used for pelagic species. A short term response to a standardized exposure is used as an indication of the
relative health of the organisms. A geometric  dilution series of five unreplicated concentrations is used
plus a negative (dilution-water only) control. Although nominal concentrations are usually sufficient for
reference toxicant tests, concentrations should be measured whenever  possible. The concentration range
should be selected to give greater than 50% mortality in at least one concentration and less than 50%
mortality in at least one concentration. An initial  range-finding test using  a  very  wide range of
concentrations may be necessary to determine the proper concentration range for reference toxicant tests.
For each species, mortality is determined and the LCjo or  EC^ is calculated as described in Appendix
D.

A control chart should be developed for each reference toxicant/test organism combination used (e.g.,
see EPA, 1990). The LC^ or ECjo for each combination should be determined  on a regular basis, and
each combination tracked on  a separate Average Control Chart (Figure G.I). Successive toxicity values
should be plotted and examined to determine if the results are within  the established limits. Commonly
used limits are the mean ±2 standard deviations. A minimum of five data points are necessary  to develop
the first  set of limits. These limits are recalculated for each  successive data  point, until the statistics
stabilize. Organisms are suitable for dredged material testing if results of reference toxicant testing fall
                                                                                 DRAFT

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  I
  8
  a
                                           G-14
             Control Chart:  Rainbow TrouVSodium Pentachlorophenate
                                    Upper Control Limit
                                    Lower Control Limit
                                             Date
              Control Chart:  Daphnia Magna/Sodium Dodecyl Sulphate
                                    Upper Control Limit
                                    Lower Control Limit
h

cp
)
. V
^ -1
d>4

k

) c
0 H
J
t * d)
                                                                                Mean
Mean
                                             Date
Figure G.I     Example Control Charts for Reference Toxicants.
                                                                            DRAFT

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	G-15	

within these limits. Outliers, or data which fall outside the upper and lower limits, suggest an atypical
population.  It is inappropriate  to use  that group  of organisms for dredged material testing  as the
sensitivity of the organisms and the overall credibility of the test system would be suspect. Reference
toxicant tests should be conducted at least monthly on each species cultured in-house, and should be
performed on each lot of purchased or field-collected organisms. The basic concept and application of
reference  toxicant tests is discussed by Lee (1980).  When sufficient reference toxicant data have been
generated for a particular species, it may be possible to specify an acceptable LC» or EC*, range for that
species with that reference toxicant.
G.2.10.5.3                    Acceptability of Test Results and Data Evaluation

For the test results to be acceptable, mean control survival must be ^ 90% or the appropriate value for
a particular test and end-point.  If mean mortality is greater than 10% or the appropriate value for a
particular test or endpoint in the control treatment for a particular test species, the test should be rejected
and repeated. Unacceptable control mortality indicates that the organisms are being affected by stress
other than contamination in the material being tested. Such  stress  may  be due to injury or disease,
unfavorable physical or chemical conditions in the test containers, improper handling or acclimation or
possibly unsuitable or contaminated water. The potential effects of these and other variables should be
carefully examined if the test is  repeated.

An individual test may be conditionally acceptable if temperature, DO, and other specified conditions fall
outside specifications. This depends on the degree of the departure and the objectives of the tests. The
acceptability of the test will depend on the experience and professional judgment of the laboratory analyst
and the reviewing staff of the regulatory authority. Any deviation from test specifications must be noted
when reporting data from a test.
G.2.11               Performance and System Audits

Audits include a careful evaluation of both field and laboratory QC procedures. They are an essential part
of the field and laboratory QA program and consist of two basic types: performance audits and system
audits. For example,  analyses of performance evaluation samples may simply be used for comparison
with the results of independent laboratories (a form of performance audit), or comprehensive audits may
be conducted by the government of the entire field or laboratory operations (a system audit).

Performance and  system  audits should be conducted  by individuals  not  directly involved  in  the
measurement process. A performance auditor independently collects data using performance evaluation
                                                                                 DRAFT

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	G-16	

samples, field blanks, trip blanks, duplicate samples, and spiked samples. Performance audits may be
conducted soon after the measurement systems begin generating data. They may be repeated periodically
as required by  task needs, duration, and cost. EPA (1991b)  should  be reviewed  for auditing the
performance of laboratories performing aquatic toxicity tests.

A systems  audit consists of a review of the total data production process. It includes on-site reviews of
field and laboratory operational systems. EPA and/or USAGE will develop and conduct external system
audits based on the approved project plan. An example of a systems audit checklist is provided in EPA
(1994).
G.2.11.1             Pre-award Inspections

The pre-award inspection is a type of system audit for assessing the laboratory's overall capabilities. This
assessment includes a determination that the laboratory personnel are appropriately qualified and that the
required equipment is available and is adequately maintained. It establishes the groundwork necessary to
ensure that tests will be conducted  properly, provides the initial contact between government and
laboratory staff, and emphasizes the importance that government places on quality work and products.
The purpose of the pre-award inspection is to verify the following:

        •      The laboratory has an independent QA/QC program.

        •      Written work plans are available for each test that describe the approach to be used in
               storing, handling, and analyzing samples.

        •      Technically  sound,  written standard operating procedures (SOPs) are available for  all
               study activities.

        •      Qualifications and training of staff are appropriate and documented.

        •      Approved analytical procedures are being followed.
 G.2.11.2              Intel-laboratory Comparisons (Chemical Analytical Laboratories)

 It is important that data collected and processed at various laboratories be comparable. As part of the
 performance audit process,  laboratories  may be  required to participate in analysis  of performance
 evaluation  samples related  to  specific  projects.  In  particular,  laboratory  proficiency  testing  is
                                                                                 DRAFT

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	G-17	

recommended. Laboratory proficiency must be demonstrated before a laboratory negotiates a contract and
yearly thereafter. Each laboratory participating in a proficiency test is required to analyze samples
prepared to a known concentration. Analytes used in preparation of the samples must originate from a
recognized source of standard reference material (SRM), such as the National Institute for Standards and
Technology (NIST). Proficiency testing programs  already established by either EPA or the USAGE may
be used, or a program may be designed specifically for dredged material evaluations. Analytical results
are compared with predetermined criteria of acceptability.

In addition, the performance evaluation samples prepared by EPA Environmental Monitoring and Systems
Laboratory (Las Vegas, Nevada)  for the Contracts Laboratory  Program (CLP) may be used to assess
interlaboratory comparability. Analytical results are compared with predetermined criteria of acceptability
(e.g., values  that fall within the  95 percent confidence interval are considered  acceptable). The QA
project plan should indicate, where applicable, scheduled participation in all interlaboratory calibration
exercises.

Reference materials are substances with well-characterized properties that are useful for assessing the bias
of an analysis and auditing analytical performances among laboratories. SRMs are  certified reference
materials containing precise concentrations of chemicals, accurately determined by a variety of technically
valid procedures, and are issued by the National Institute of Standards and Technology. Currently, SRMs
are not available for the  physical measurements  of all contaminants in  sediments; however,  where
possible, available SRMs or other regional reference materials that have been repeatedly tested should
be analyzed with every 20 samples processed.

SRMs for most organic compounds are not currently available for seawater, but reference materials for
many inorganic chemicals  may be obtained from the organizations listed in Table G.I. Seawater matrix
spikes of target analytes (e.g., seawater spiked with National Institute for Standards and Technology SRM
1647 for PAH) should be used to check analytical bias. Some available SRMs for priority pollutant metals
in seawater are National Research Council of Canada seawater CASS-1 and seawater NASS-2.

SRMs for organic priority pollutants in tissues are currently not available. The National Institute of
Standards and Technology  is presently developing SRMs for organic analytes. Tissue matrix spikes of
target analytes should be used to fulfill analytical  accuracy requirements  for organic  analyses.

Because new SRMs appear constantly, current listings of appropriate  agencies should  be consulted
frequently. SRMs that are readily available and commonly used are included in Table G.I.
                                                                                DRAFT

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Table G.I
                                           G-18
       Sources of Standard Reference Materials
 PCBs
 PAHs
National Research Council of Canada   Marine sediment


National Research Council of Canada   Marine sediment

                                   Sediment
       National Institute for Standards and
       Technology
 Metals
       National Bureau of Standards
                                   Estuarine sediment
       National Research Council of Canada  Marine sediment


                                          Dogfish liver

                                          Dogfish muscle

                                          Lobster hepatopancreas

       International Atomic Energy Agency   Marine sediment

                                          Fish flesh

                                          Mussel tissue
                                                                  HS-1 and HS-2
HS-3, HS-4, HS-5, HS-6

SRM #1647 and SRM
#1597
SRM #1646

MESS-1, BCSS-1,
PACS-1

DOLT-1

DORM-1

TORT-1

SD-N-1/2(TM)

MA-A-2(TM)

MAL-1(TM)
Standard reference materials (SRMs) may be obtained from the following organizations:
 Organic Constituents

    U.S. Department of Commerce
    National Institute for Standards of
    Technology
    Office of Standard Reference Materials
    Room B3111 Chemistry Building
    Gaithersburg, Maryland  20899
    Telephone: (301) 975-6776

 Inorganic Constituents

    U.S. Department of Commerce
    National Institute for Standards and
    Technology
    Office of Standard Reference Materials
    Room B3111 Chemistry Building
    Gaithersburg, Maryland  20899
    Telephone: (301) 975-6776
                                       Marine Analytical Chemistry Standards Program
                                       National Research Council of Canada
                                       Atlantic Research Laboratory
                                       1411 Oxford Street
                                       Halifax, Nova Scotia, Canada B3H 3Z1
                                       Telephone: (902) 426-8280
                                       Marine Analytical Chemistry Standards Program
                                       National Research Council of Canada
                                       Division of Chemistry
                                       Montreal Road
                                       Ottawa, Ontario, Canada K1A OR9
                                       Telephone: (613) 993-2359
                                                                            DRAFT

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	G-19	

G.2.11 J              Routine Inspections

Routine system audits during the technical evaluation ensure that laboratories are complying with the QA
project plan. It is suggested that checklists be  developed for  reviewing training records, equipment
specifications,  QC procedures  for analytical tasks, management organization, etc. An example of a
systems audit is provided in EPA (1994). Districts should also establish laboratory review files for quick
assessment of the laboratory's  activity on a study,  and to aid in monitoring the overall quality of the
work. Procedures for external systems audits by the Districts are similar to the internal systems audits
conducted by the laboratories themselves.
G.2.12               Facilities

The QA Project  Plan should provide a complete, detailed description of the physical  layout of the
laboratory, define space for each test area, describe traffic-flow patterns, and document special laboratory
needs. The design and layout of laboratory facilities are important to maintain  sample integrity and
prevent cross-contamination. The specific areas  to be  used for the various evaluations  should  be
identified. Aspects of the dredging study that warrant separate facilities include the following:

       •      receiving
       •      sample storage
       •      sample preparation
       •      sample testing
       •      reagent storage
       •      data reduction and analysis.
G.2.13               Preventive Maintenance

The QA project plan should describe how field and laboratory equipment essential to sample collection
and analysis will be maintained in proper working order. Preventive maintenance may be in the form of:
1) scheduled maintenance activities to minimize costly downtime and ensure accuracy of measurement
systems, and 2) available spare parts, backup systems, and equipment. Equipment should be subject to
regular inspection and preventive maintenance procedures to ensure proper working order. Instruments
should have periodic calibration and preventive maintenance performed by qualified technical personnel,
and a permanent  record kept of calibrations, problems diagnosed, and corrective actions applied. An
acceptance testing program for key materials used in  the performance of environmental measurements
(chemical and  biological materials) should be applied prior to their use.
                                                                                 DRAFT

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	G-20	

G.2.14                Calculation of Data Quality Indicators

The calculations and  equations used  routinely in QA  review (e.g., relative percent difference of
duplicates) as well as the type of samples (e.g., blanks, replicates) analyzed to assess precision, bias, and
completeness of the data must be presented in the QA project plan. Routine procedures for measuring
precision and  bias include use of replicate analyses, standard reference materials, and matrix spikes.
Completeness  can be measured for each  set of data  received by dividing the number of valid (i.e.,
accepted) measurements actually obtained  by the number of measurements that were planned.
G.2.15                Corrective Actions (Management of Nonconformance Events)

One purpose of any QA program is to identify nonconformance as quickly as possible. A nonconformance
event is defined as any event that does not follow defined methods, procedures, or  protocols, or any
occurrence that may affect the quality of the data or study.  A QA program should  have a corrective
action plan and  should provide  feedback  to appropriate management  authority  defining how all
nonconformance events were addressed and corrected.

Corrective actions fall into two categories:  1) handling of analytical or equipment malfunctions, and 2)
handling of nonconformance or noncompliance with the QA requirements that have been established.
During field  and  laboratory  operations,  the supervisor  is responsible for  correcting  equipment
malfunctions. All corrective measures taken must be documented and, if required, an alteration checklist
must  be completed.

Corrective action procedures must be described for each project and include the following elements:

       •       procedures  for corrective actions  when predetermined limits for data acceptability are
               exceeded (see "data quality objective" discussion in Section G.2.3)

       •       for  each measurement  system, identify  the individual responsible  for  initiating the
               corrective action and also the individual responsible for approving the corrective action.

Corrective actions may be initiated as a result of other QA activities including performance audits, system
audits, interlaboratory/interfield comparison studies, and QA program audits. An example of a corrective
actions checklist is provided in Appendix G.4.
                                                                                 DRAFT

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	G-21	

G.2.16               QA Reports to Management

QA Project Plans provide a mechanism for periodic reporting to management on the performance of
measurement systems and data quality. At a minimum, these reports should include:

       •      periodic assessment of measurement data accuracy (precision and bias), and completeness

       •      results of performance and system audits

       •      significant QA problems and recommended solutions.

The individuals responsible for preparing the periodic reports should be identified. The final report for
each project must include a separate QA section which summarizes data quality information contained
in the periodic reports.


G.3           REFERENCES

EPA. 1987. Quality Assurance/Quality Control (QA/QC)for 301 (h) Monitoring Programs: Guidance on
       Field and Laboratory Methods. EPA 430/9-86-004. Prepared by Tetra Tech, Inc., Bellevue WA,
       for the U.S. Environmental Protection Agency Office of Marine and Estuarine Protection. NTIS
       Number PB87-221164.

EPA. 1990. Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater
       and Marine Organisms. EPA/600/4-90/027. U.S. Environmental Protection Agency, Office of
       Research and Development, Washington, D.C. 293 pp.

EPA. 199la. A Project Manager's Guide to Requesting and Evaluating Chemical Analyses. Prepared by
       PTI Environmental Services, Bellevue, WA, for the  U.S. Environmental Protection Agency
       Region 10, Puget Sound Estuary Program, Seattle,  WA.

EPA. 1991b. Manual for the Evaluation of Laboratories Performing Aquatic Toxicity Tests. EPA/600/4-
       90/031.  U.S.  Environmental  Protection  Agency, Office of Research  and  Development,
       Washington, D.C.

EPA. 1991c. Technical Support Document for Water Quality-Based Toxics Control. EPA/505/2-90-001.
       U.S. Environmental Protection Agency, Office of Water, Washington, D.C.
                                                                            DRAFT

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	G-22	

EPA. 1994. QA/QC Guidance for Sampling and Analysis of Sediments, Water, and Tissues for Dredged
       Material Evaluations. Phase I - Chemical Evaluations. In Press. U.S. Environmental Protection
       Agency, Office of Water, Washington, D.C.

EPA/USACE. 1991. Evaluation of Dredged Material Proposed for Ocean Disposal - Testing Manual.
       EPA-503/8-91/001, Washington, DC.

Lee, D.R. 1980. Reference toxicants in quality control of aquatic bioassays. Pp. 188-199 In: A.L.
       Buikema, Jr. and J. Cairns,  Jr. (Eds.), Aquatic Invertebrate Bioassays. ASTM STP 715.
       American Society for Testing and Materials, Philadelphia, PA.

Sturgis, T.C. 1990. Guidance for contracting biological and chemical evaluations of dredged material.
       Tech. Kept. D-90-XX, U.S. Army Engineer Waterways Experiment Station, Vicksburg, MS.

Wang, W. 1987. Chromate ion as a reference toxicant for aquatic phytotoxicity tests. Environ. Toxicol.
       Chem. 6: 955-960.
                                                                             DRAFT

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         G-23
     APPENDIX G.4
    EXAMPLE QA/QC
CHECKLISTS, FORMS, AND
       RECORDS
                             DRAFT

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                               G-24
                      TABLE OF CONTENTS
                                                           Page No.
QA PROGRAM ORGANIZATION FLOW DIAGRAM                         G-25

EXAMPLE DATA QUALITY OBJECTIVES FOR
PRECISION, ACCURACY, AND COMPLETENESS                           G-26

ALTERATION CHECKLIST                                          G-27

GENERAL SAMPLE LABEL                                          G-28

FIELD TRACKING REPORT FORM                                    G-29

LABORATORY TRACKING REPORT FORM                              G-29

CHAIN-OF-CUSTODY RECORD                                       G-30

STATION LOCATION LOG                                          G-31

CORRECTIVE ACTIONS CHECKLIST                                   G-32
                                                       DRAFT

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                                   G-25
i
i
                QA PROGRAM ORGANIZATION FLOW DIAGRAM
                      PROGRAM MANAGER
       REGULATORY
         OFFICER
REGULATORY
 OFFICER
REGULATORY
 OFFICER
                          PROJECT
                          MANAGER
                        ASSISTANT
                      PROJECT MANAGER
                          PROJECT
                      QA COORDINATOR
      QA CHEMISTRY
 QA DATA
 ANALYSIS
                                                              DRAFT

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                              	G-27	
                               ALTERATION CHECKLIST
Sample Program Identification:
Material to be Sampled: 	
Measurement Parameter:  	
Standard Procedure for Analysis:
Reference:
Variation from Standard Procedure:
Reason for Variation:
Resultant Change in Field Sampling Procedure:
Special Equipment, Material, or Personnel Required:
Author's Name:	Date:
Approval:  	Tide:
Date:	
                                                                          DRAFT

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     	G-28	




     GENERAL SAMPLE LABEL
(NAME OF SAMPLING ORGANIZATION)




PROJECT:  	




DATE:  	




TIME:   	
SAMPLE ID NO.:




MEDIA:  	
STATION NUMBER:




DEPTH:  	
PRESERVATION:
ANALYSES TO BE PERFORMED^




SAMPLED BY: 	




LAB NO.:  	




REMARKS:  	
                                       DRAFT

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              G-29
   FIELD TRACKING REPORT FORM
W/O No.


FIELD TRACKING REPORT:

FIELD SAMPLE
CODE
(FSC)




BRIEF
DESCRIPTION



(LOC-SN)
DATE



TIME



Page

SAMPLER



LABORATORY TRACKING REPORT FORM
W/O No.


LABORATORY TRACKING REPORT:

FRACTION CODE



X




PREP/ANAL
REQUIRED



(LOC-SN)
RESPONSIBLE
INDIVIDUAL



DATE
DELIVERED



Page

DATE
COMPLETED



                                     DRAFT

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                                    G-31
                          STATION LOCATION LOG
                                                        DATE:
PROJECT:
STATION LOCATION:
DESCRIPTION OF SAMPLES COLLECTED:
SPC ZONE: 	(N/S)      EAST:
                           NORTH:
LOCATION:
Bottom Depth:	(ft)
LORANC:  LOP1
         (m)      Tide: ±
           LOP2	
       (m) MLLW:
       (ft)
(m)
Variable Radar Range:
Visual Fixes: (Note:  Please tape any drawings to back of this sheet)
Photos - Roll:
 Pictures:
PID Reading (range):

Comments: 	
RECORDER:
SIGNATURE:
ORG. CORE
DATE:
                                                               DRAFT

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                                 G-32



                    CORRECTIVE ACTIONS CHECKLIST




SAMPLE PROGRAM IDENTIFICATION:  	




SAMPLING DATES: 	
t
MATERIAL TO BE SAMPLED:




MEASUREMENT PARAMETER:




ACCEPTABLE DATA RANGE:
CORRECTIVE ACTIONS INITIATED BY: 	




TITLE: 	




DATE: 	




PROBLEM AREAS REQUIRING CORRECTIVE ACTION:
MEASURES TO CORRECT PROBLEMS:
MEANS OF DETECTING PROBLEMS (FIELD OBSERVATIONS, SYSTEMS AUDIT, ETC):
APPROVAL FOR CORRECTIVE ACTIONS:




TITLE: 	
SIGNATURE:
DATE: 	                  »D.S. GOVERNMENT PRINTING OFFICE: 1994-533-246/00003

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