&EPA
United States
Environmental Protection
Agency
Office of Research and
Development
Washington, DC 20460
EPA/600/R-92/111
March 1993
Field and
Laboratory Methods for
Evaluating the Biological
Integrity of Surface Waters
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EPA/600/R-92/111
March 1993
FISH FIELD AND LABORATORY METHODS FOR EVALUATING
THE BIOLOGICAL INTEGRITY OF SURFACE WATERS
Donald J. Klemm1, Quentin J. Stober2, and James M. Lazorchak1
1Bioassessment and Ecotoxicology Branch,
Ecological Monitoring Research Division
Environmental Monitoring Systems Laboratory - Cincinnati, Ohio
2Ecological Support Branch, Environmental Services Division -
Region IV, Athens, Georgia
ENVIRONMENTAL MONITORING SYSTEMS LABORATORY - CINCINNATI
OFFICE OF MODELING, MONITORING SYSTEMS, AND QUALITY ASSURANCE
OFFICE OF RESEARCH AND DEVELOPMENT
U. S. ENVIRONMENTAL PROTECTION AGENCY
CINCINNATI, OHIO 45268
y Printed on Recycled Paper
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DISCLAIMER
This document has been reviewed by the Environmental Monitoring Systems
Laboratory - Cincinnati (EMSL-Cincinnati), U.S. Environmental Protection
Agency (USEPA), and approved for publication. The mention of trade names or
commercial products does not constitute endorsement or recommendation for use.
11
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FOREWORD
Environmental measurements are required to determine the quality of
ambient waters and the character of waste effluents. The Environmental
Monitoring Systems Laboratory - Cincinnati (EMSL-Cincinnati) conducts research
to:
o Develop and evaluate methods to identify and measure the
concentration of chemical pollutants in drinking waters, surface
waters, groundwaters, wastewaters, sediments, sludges, and solid
wastes.
o Investigate and evaluate methods for the identification and
measurement of viruses, bacteria and other microbiological organisms
in aqueous samples and to determine the response of aquatic organisms
to water quality.
o Perform ecological assessments and measure the toxicity of pollutants
to representative species of aquatic organisms and determine the
effects of pollution on communities of indigenous freshwater,
estuarine, and marine organisms, including the phytoplankton,
zooplankton, periphyton, macrophyton, macroinvertebrates, and fish.
o Develop and operate a quality assurance program to support the
achievement of data quality objectives in measurements of pollutants
in drinking water, surface water, groundwater, wastewater, sediment
and solid waste.
o Develop methods and models to detect and quantify responses in
aquatic and terrestrial organisms exposed to environmental stressors
and to correlate the exposure with effects on biochemical and
biological indicators.
This manual describes guidelines and standardized procedures for the use
of fish in evaluating the biological integrity of surface waters. It was
developed to provide biomonitoring programs with fisheries methods for
measuring the status and trends of environmental pollution on freshwater,
estuarine, and marine habitats in field and laboratory studies. These fish
studies are carried out to assess biological criteria for the recognized
beneficial uses of water, to monitor surface water quality, and to evaluate
the health of the aquatic environment.
Thomas A. Clark
Director
Environmental Monitoring Systems
Laboratory - Cincinnati
m
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PREFACE
The Bioassessment and Ecotoxicology Branch, Ecological Monitoring
Research Division, Environmental Monitoring Systems Laboratory - Cincinnati is
responsible for the development, evaluation, and standardization of methods
for the collection of biological field and laboratory data by EPA regional,
enforcement, and research programs engaged in inland, estuarine, and marine
water quality and permit compliance monitoring, and status and/or trends
monitoring for the effects of impacts on aquatic organisms, including the
phytoplankton, zooplankton, periphyton, macrophyton, macroinvertebrates, and
fish. The program addresses methods for sample collection; sample
preparation; organism identification and enumeration; the measurement of
biomass and metabolic rates; the bioaccumulation and pathology of toxic
substances; bioassay; biomarkers; the computerization, analysis, and
interpretation of biological data; and ecological assessments.
This manual contains field and laboratory fish methods for evaluating
the health and biological integrity of fresh, estuarine, and marine waters.
The manual is a revision and enlargement of the chapter on fish methods
originally published in the document, "Biological Field and Laboratory Methods
for Measuring the Quality of Surface Waters and Effluents," Environmental
Monitoring Series, USEPA, 1973, EPA-670/4-73-001, which were developed by the
Bioassessment and Ecotoxicology Branch, Environmental Monitoring Systems
Laboratory - Cincinnati, at the request of the Biological Advisory Committee
to provide biomonitoring programs with methods for assessing point and
nonpoint sources of impacts, status and trends in water quality monitoring.
IV
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ABSTRACT
This manual contains biocriteria and describes guidelines and
standardized methods for using fish in evaluating the health and biological
integrity of surface waters and for protecting the quality of water resources.
Included are sections on quality assurance and quality control procedures;
safety and health recommendations; fish collection techniques; specimen
processing techniques; identification and taxonomic references; fish age,
growth, and condition determinations; data recording; length-frequency;
length-age conversion; annulus formulation; relative weight index; flesh
tainting; fish kill investigation; bioassessment protocols for use in streams
and rivers; family-level ichthyoplankton index; fish health and condition
assessment; guidelines for fish sampling and tissue preparation for
bioaccumulative contaminants; and an extensive bibliography for fisheries.
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CONTENTS
Foreword. , ....... ...... iii
Preface iv
Abstract. ..................... v
Figures ......... x
Tables xiii
Acknowledgment. .......... xvi
1. Introduction. ...... 1
Literature Cited. ..... 7
2. Quality Assurance and Quality Control .... 15
Introduction. ....... ... 15
Data Quality Objectives ........ 16
Facilities and Equipment 18
Calibration, Documentation, and Record Keeping 19
Habitat Assessment. 20
Fish Collection ...... 22
Qualification and Training. 22
Standard Operating Procedures . 23
Literature Cited. ................ 24
3. Safety and Health ...... ..... 27
Introduction. ................ 27
General Precautions ........... 27
Safety Equipment and Facilities .............. 28
Field and Laboratory Operations ........ 29
Disease Prevention. ....... ...... 29
Literature Cited. ..................... 29
4. Sample Collection for Analysis of the Structure and
Function of Fish Communities. 31
General Considerations. ............. 31
Habitat Evaluation. ...... 34
Active Sampling Techniques. .... 42
Seines. .. T 42
Trawls. 44
Horizontal Ichthyoplankton Tow-net. ........... 47
Electrofishing. ..... 49
Chemical Fishing (Ichthyocides) 56
Hook and Line 59
Passive Sampling Techniques ...... .... 59
Entanglement Nets ............ 62
Entrapment Devices 63
Pop Nets. 67
vi
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CONTENTS (CONTINUED)
Miscellaneous Fish Methods 68
Underwater Methods 68
Hydroacoustic Techniques 68
Underwater Biotelemetry 68
Literature Cited. ... 69
5. Specimen Processing Techniques 78
Introduction 78
Fixation and/or Preservation of Fish Samples. ....... 78
Labelling of Specimens in Field and Laboratory 80
Species Identification 80
Literature Cited 82
6. Sample Analysis Techniques 83
Introduction. 83
Data Recording 83
Fish Identification 84
Species Composition (Richness) 84
Length and Weight 85
Age, Growth, and Condition 8i
Length-frequency Method 87
Length-Age Conversion Method 87
Annul us Formation Method 90
Condition Factor (Coefficient of Condition) 91
Relative Weight Index ..... 92
Literature Cited 93
7. Special Techniques 96
Flesh Tainting 96
Fish Kill Investigations. 96
Instream Flow Incremental Methodology (IFIM) 121
Fish Marking and Tagging Techniques ............ 122
Literature Cited 122
8. Fish Bioassessment Protocols For Use In Streams and Rivers. . 128
Introduction. ........ . 128
Sampling Representative habitat .... 133
Fish Sample Processing and Enumeration 133
Fish Environmental Tolerance Characterizations 134
Fish Biosurvey and Data Analysis 134
USEPA Fish Bioassessment I. ........ 142
USEPA Fish Bioassessment II 147
Description of IBI Methods . 154
Guidance for Use of Field Data Sheets 163
Guidance for Impairment Assessment Sheet 166
Guidance for Field Collection Data Sheet for Fish
Bioassessment II, ... ..... 166
Vll
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CONTENTS (CONTINUED)
Guidance for Data Summary Sheet for Fish Bioassessment
II 167
Habitat Assessment, Physical and Chemical Parameters. . . . 168
Physical Characteristics and Water Quality 168
Habitat Quality and Assessment 172
Selected References for Determining Fish Tolerances,
Trophic, Reproductive, and Origin Classification 182
Agencies Currently using or Evaluating Use of the IBI
or Iwb for Water Quality Investigations 192
Ohio EPA Fish Index of Biotic Integrity (IBI),
Modified Index of Well-Being (Iwb), and Qualitative
Habitat Evaluation Index (QHEI) 193
Literature Cited 198
9. Family-Level Ichthyoplankton Index Methods 205
Introduction 205
Methods and Materials 210
Taxonomic Considerations 226
Provisional Key to the Families of North American
Freshwater Fishes 228
Fish Larvae Sampling Precision 231
Literature Cited 232
10. Fish Health and Condition Assessment Profile Methods 239
Introduction 239
Sampling and Collection of Fish 241
Handling of Fish 241
Sampling and Reading of Blood 241
Length and Weight Measurements 242
External Examination 242
External Organs 243
Internal Examination (or Necropsy) 250
Calculation and Summary of Fish Health and Condition
Assessment 255
AUSUM 2.6--Computer Program for the Necropsy-Based Fish
Health and Condition Assessment System 261
Literature Cited 288
11. Guidelines for Fish Sampling and Tissue Preparation
for Bioaccumulative Contaminants 289
Introduction 289
Site Selection 290
Sample Collection 290
Sample Preparation for Organic Contaminants in Tissue . . . 294
Sample Preparation for Metal Contaminants in Tissue .... 300
Identification of Composite Whole Fish or Fillet Samples. . 301
Chain-of-Custody Procedures 302
Conclusion 303
Literature Cited 303
viii
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CONTENTS (CONTINUED)
12. Fisheries Bibliography 305
General References 305
Electrofishing 318
Chemical Fishing 322
General Health, External Anomalies, Deformities,
Eroded Fins, Parasites, and Diseases 324
Fish Identification 326
General 326
Larval and Immature Fishes . 329
Marine: Atlantic and Gulf of Mexico 330
Marine: Coastal Pacific 333
Freshwater: Northeast 334
Freshwater: Southeast 336
Freshwater: Midwest 338
Freshwater: Southwest 340
Freshwater: Northwest 342
Canada 344
Fish Kills. 345
IX
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FIGURES
SECTION 2
Number Page
1. Example of sample identification tag 20
2. Example of a chain-of-custody record form 21
SECTION 4
Number Page
1. General fish field data sheets 35
2, Site description sheet for evaluating the topogeographical
features and physical characteristics of fish sampling location. . . 39
3, Common Haul seine 45
4, Beam trawl 45
5. Otter trawl 46
6. Horizontal ichthyoplankton tow-net 48
7. Boom shocker 50
8. Gill net 63
9. Trammel net 65
10. Hoop net. 65
11. Fyke net 66
12. Slat trap ............................. 66
13. Pop net 67
SECTION 5
Number Page
1. Examples of field sample data labels 81
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FIGURES (CONTINUED)
SECTION 6
Number Page
1. Example of fish sample label information for preserved
specimen container. .................. 84
2. Fish measurements (using a fish measuring board)
and scale sampling areas. ............ ... 86
3. Example of recording field data information of scale
samples for age and growth studies. ..... 90
SECTION 7
Number Page
1. Minimum water sampling point on stream 200 feet or less
wide involving an isolated discharge ........... 117
2. Minimum water sampling points on a stream running through
an industrial or municipal complex .......... 117
SECTION 8
Number Page
1. Flowchart of biosurvey approach for fish bioassessment II. .... 141
2. Range of sensitivities of biosurvey for fish bioassessment
II metrics in assessing biological condition 142
3. Fish assemblage questionnaire for use with fish bioassessment I. . 144
4. Impairment assessment sheet for use with fish bioassessment II . . 149
5. Fish field collection data sheet for use with fish bioassessment
II 151
6. Total number of fish species versus watershed area for Ohio
regional reference sites ............... 159
7. Data summary sheet for fish bioassessment II ........... 164
8. Header information used for documentation and identification
for sampling stations. ............. 165
9. Physical characterization/water quality field data sheet for
use with bioassessment ............... 169
xi
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FIGURES (CONTINUED)
SECTION 8 (CONTINUED)
Number Page
10. Habitat assessment field data sheet, riffle/run prevalence 173
11. Habitat assessment field data sheet, glide/pool prevalence 174
12. Example of Ohio EPA (1991) quantitative habitat evaluation
index field sheet 175
13. Flowchart of biosurvey approach for fish bioassessment
used by Ohio EPA (1991) 194
14. Example of Ohio EPA (1991) field data sheet constructed
for immediate entry into a computer data base 196
SECTION 9
Number Page
1. Morphometric characteristics of larval fish 227
2. Diagrammatic representation of morphology of a teleost larva. , . . 227
SECTION 10
Number Page
1. External features of a composite fish 244
2. Fish necropsy work sheet 247
3. Anatomy of a soft-rayed bony fish, the brook trout,
SalveTinus fontinalis ....... 252
4. Anatomy of a spiny-rayed bony fish, the largemouth bass,
Micropterus saTmoides ..... 253
SECTION 11
Number Page
1. General sampling scheme for bioaccumulative contaminant
in fish, multiple age groups will require additional samples. . . . 295
xn
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TABLES
SECTION 1
Number Page
1. Attributes of fishes and desirable components for
bioassessments and biomonitoring programs 3
2. Five major classes of environmental factors which
influence and determine the biological integrity of
surface waters with some of their important chemical,
physical, and biological components in lentic and
lotic systems 4
SECTION 2
Number Page
1. Example of summary table for data quality requirements 18
SECTION 4
Number Page
1. General indicators of biological/ecological integrity for fish. . . 32
2. General checklist of fish field equipment and supplies 33
3. Codes utilized to record external anomolies on fish 41
4. Amount of 5% emulsifiable rotenone equivalent to 0.5 ppm
or 1.0 ppm per acre-feet or pond or lake to be sampled 60
5. Cubic centimeters (cc) of liquid rotenone per minute for
gallons of flow per minute 61
SECTION 5
Number Page
1. Formulation of formalin fixative solution 79
SECTION 6
Number Page
1. Average total lengths in inches for each age group of several
fishes in Michigan 89
xm
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TABLES (CONTINUED)
SECTION 7
Number Page
1. Flowchart for the coordination of a fish kill investigation .... 100
2. Fish kill general information form 103
3. Checklist of fish kill investigation equipment. 105
4. Field observations 106
5. Fish kill investigation form 107
6. Observations on dead and moribund fish 109
7. Observations on affected fish Ill
8. Symptoms that have been related to cause of fish death 113
9. Summary of a lower Mississippi River endrin fish kill
investigation 120
SECTION 8
Number Page
1, Tolerance designations, trophic status, and North American
endemicity of selected fish species 135
2. Regional variations of IBI metrics. . 156
3. Nine habitat parameters and assessment category 178
SECTION 9
Number Page
1. Taxonomic literature useful for identification of larval
and early juvenile North American freshwater fish 206
2. Total ichthyoplankton index (I2) scores, integrity classes,
and attributes 209
3. Metrics used to assess ichthyoplankton communities from
freshwater from North America 213
xiv
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TABLES (CONTINUED)
SECTION 9 (CONTINUED)
Number Page
4. Sensitivities, mean generation time, and reproductive
guild characteristics of 34 North American fish families 214
5. The diversity of species, d, characteristics of MacAuthur's
model for various numbers of hypothetical species, S' 216
6. Classification of reproduction styles for fishes in order
of evolutionary trends 218
SECTION 10
Number Page
1. Equipment and materials for fish health and
condition assessment 240
2. Necropsy classification outline 245
3. Summary of fish necropsy 257
4. Sample of fish necropsy computer summary report I ... 262
5. Sample of fish necropsy computer summary report II 264
SECTION 11
Number Page
1. Frequency of occurrence for freshwater and marine species
in the national fish bioaccumulation study (USEPA, 1990a) 292
2. Summary of sample collection and preparation QA/QC
requirements for fish tissue (Modified from Puget
Sound estuary program, 1986 and 1989) 296
xv
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ACKNOWLEDGMENTS
The subcommittee for fish, John Hale, Paul Frey, Ernest Karvelis, James
LaBuy, Loys Parrish, Ronald Preston, and Richard Wagner are recognized as
first contributors to the fish chapter, 19 pages, in the USEPA, 1973,
"Biological Field and Laboratory Methods for Measuring the Quality of Surface
Waters and Effluents," edited by Cornelius I. Weber.
Technical review comments from the following individuals are gratefully
acknowledged:
Michael T. Barbour, Tetra Tech, Owings Mills, MD
Donald Brockway, USEPA, Region 4, Environmental Services Division, Athens, 6A
Philip A. Crocker, USEPA, Region 6, Water Quality Management Branch, Dallas,
TX
Eric Dohner, Tetra Tech, Owings Mills, MD
Robert Donaghy, USEPA, Region 3, Wheeling Office, Wheeling, WV
Janet Kuefler, USEPA, Region 9, Water Management Division, San Francisco, CA
Philip A. Lewis, USEPA, EMSL, Bioassessment and Ecotoxicology Branch,
Cincinnati, OH
Robert Nester, U.S. Fish and Wildlife Service, Great Lakes Fisheries
Laboratory, Ann Arbor, MI
Peter Nolan, USEPA, Region 1, New England Regional Laboratory, Lexington, MA
Loys Parrish, USEPA, Region 8, Environmental Service Division, Denver, CO
Quentin H. Pickering, USEPA, EMSL, Bioassessment and Ecotoxicology Branch,
Cincinnati, OH
Thomas P. Simon, USEPA, Region 5, Environmental Services Division, Chicago, IL
Mark Smith, Technology Applications, Inc., Cincinnati, OH
Sam Stribling, Tetra Tech, Owings Mills, MD
Betsy Sutherland, USEPA, Standard and Applied Science Division, Washington, DC
William Sutton, USEPA, Region 4, Environmental Services Division, Athens, GA
Irene M. Suzukida, USEPA, Water Quality and Industrial Permitting Branch,
Washington, DC
William Thoney, Technical Applications, Inc., Cincinnati, OH
Cornelius I, Weber, USEPA, Ecological Monitoring Research Division,
Cincinnati, OH
Roger Yeardly, Technology Applications, Inc., Cincinnati, OH
Chris Yoder, Ohio EPA, Columbus, OH
We especially thank Ronald W. Goede, Utah Division of Wildlife
Resources, for providing the fish health and condition assessment procedures.
We greatly appreciate the illustrated written computer program, AUSUM 2.6, for
the necropsy-based, fish health and condition assessment system that Ronald W.
Goede and Sybil Houghton contributed.
We are very grateful to Thomas P. Simon, Regional Biocriteria
Coordinator and State of Ohio Standards Coordinator; USEPA, Environmental
Services Division, Region 5, Chicago, IL, for his review of the technical
contents and for the information on the relative weight index and the
ichthyoplankton index.
xvi
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ACKNOWLEDGMENTS (CONTINUED)
Frank H. McCormick, USEPA, EMSL, Ecological Monitoring Research
Division, Bioassessment and Ecotoxicology Branch, Cincinnati, OH deserves
special thanks for his critical review of the technical contents of the
manual.
We acknowledge F. Bernard Daniel, Director, Ecological Monitoring
Research Division (EMRD), Environmental Monitoring Systems Laboratory (EMSL),
Cincinnati for his review of this manual.
Special thanks go to Lora Johnson, Quality Assurance Manager,
Environmental Monitoring Systems Laboratory, Cincinnati for reviewing Section
2, Quality Assurance and Quality Control; Laura Gast, Technology Applications,
Inc., Cincinnati for reviewing the statistics; and Debbie Hall, Secretary,
Bioassessment and Ecotoxicology Branch, EMRD, EMSL, Cincinnati for providing
secretarial assistance.
xvi i
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SECTION I
INTRODUCTION
1.1 This manual was prepared to assist biologists and managers in USEPA and
other Federal, state, and private water monitoring organizations in the use of
fish as indicators of ecosystem health and for evaluating the biological
integrity of surface waters and protecting quality water resources. The
manual contains biological criteria and laboratory and field methods that will
aid in the monitoring and bioassessment of the effects of anthropogenic and
environmental stresses on fish populations and communities. It will also
facilitate the expansion and refinement of our knowledge of the ecological
requirements of fish species in freshwater, estuarine, and marine habitats.
1.2 The manual includes sections on quality assurance and quality control,
safety and health, sampling methods and techniques, sample preservation and
identification, data analyses, special techniques, bioassessment protocols for
use in streams and rivers, a family-level ichthyoplankton index method, fish
health and condition assessment procedures, guidelines for fish sampling and
tissue preparation for bioaccumulative contaminants, and a fisheries
bibliography. Guidelines and procedures for fish kill investigations are
provided.
1.3 Fish community evaluation and assessment should measure the overall
structure (number of species and individuals within a community) and function
(organism interaction in the utilization of food and other biological
resources) of various aquatic habitats considered for study. These
measurements should include such factors as habitat characteristics and
quality, riparian vegetation, and hydraulic characteristics that are expected
to influence fish community spatial and temporal variability. One must also
distinguish the alterations induced by anthropogenic activities from natural
variations which occur in the environment.
1.4 In North America, fish are the focus of economically important sport and
commercial fisheries, and are an important source of food for humans. To the
general public the size and species composition of a fish community is the
most meaningful index of pollution.
1.5 In most aquatic ecosystems, fish are usually the most common vertebrates.
Fish communities occupy the upper trophic levels of aquatic food webs, and
they are dependent on the same or other trophic level life forms for food. In
aquatic communities fish can be one of the most sensitive indicators of water
quality assessment and biological integrity in aquatic environments
(Angermeier et al., 1991; Fausch et al., 1990; Karr, 1981, 1987, 1990, 1991;
Smith, 1971; McKenzie et al., 1992). The literature contains much data on
fish species distribution, life histories, ecology, pollution tolerance, and
environmental requirements. Fish are directly and indirectly affected by
chemical and physical changes in the environment, and the population or
community of fish in rivers, streams, lakes, estuaries, and oceans reflects
the state of the health of the aquatic environment or watershed as a whole.
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Because they are conspicuous, fish populations or fish assemblages are
commonly used as environmental indicators or as an index for water quality
(Table 1).
1.6 Water quality conditions that significantly affect the lower levels of
food webs (e.g., plankton and benthic invertebrates, including
macroinvertebrates, USEPA, 1990a) will affect the abundance and species
composition of the fish population. In some cases, fish may exhibit signs of
being more sensitive to certain pollutants than are the lower animals and
plants, and may be adversely affected even when the lower levels of food webs
are relatively unharmed.
1.7 Karr (1981, 1987), Karr et al. (1986, 1987), Ohio EPA (1990), and USEPA
(1990a,b) have indicated that five major sets of abiotic and biotic factors
affect and ascertain biological integrity or water resource integrity (Table
2). To determine anthropogenic or natural impact on aquatic ecosystems, all
monitoring or bioassessment programs must survey and evaluate in a methodical
and systematic way all five sets of factors. Although a thorough discussion
of all these factors is beyond the scope of this document, a discussion of how
some of these factors influence the biological integrity of surface waters and
several methods and procedures in evaluating these complex set of factors are
presented here. For a more comprehensive discussion of all these factors,
consult USEPA (1990a, 1990b), Ohio EPA (1990), and the references in Section
12, Fisheries Bibliography.
1,8 Many species of fish have stringent dissolved oxygen and temperature
requirements and are intolerant to chemical and physical contaminants
resulting from municipal, agricultural, industrial, forestry, and mining
activities. Also, fish communities are sensitive to and good indicators of
macrohabitat disturbances (Rankin, 1989).
1.9 The discharge of moderate amounts of degradable organic wastes may
increase the nutrient levels (eutrophication) in the habitat and result in an
increase in the standing crop (total amount of the biomass of organisms of one
or more species within a locality) of fish. This increase usually occurs in
one or a few species and results in an imbalance in the population. The
discharge of large amounts of degradable organic materials may result in
depressed oxygen levels which may reduce the number and kinds of fishes
present and increase the standing crop of pollution tolerant species. In
extreme cases the fishery may be eliminated in the affected area.
1.10 The effects of toxic wastes may range from the elimination of most fish
to a reduction in reproductive capacity (fecundity) or resistance to disease
and parasitism. Massive and complete fish kills are dramatic signs of abrupt,
adverse changes in environmental conditions. Fish, however, can repopulate an
area rapidly if the habitat is not destroyed and the water quality improves.
The cause of the fish kill may be difficult to detect by examination of the
fish community after it has recovered from the effects of the pollutant.
Chronic pollution, on the other hand, is more selective in its effects, exerts
its influence over a long period of time, and causes recognizable changes in
the species composition and relative abundance of the fish.
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TABLE 1. ATTRIBUTES OF FISHES AND DESIRABLE COMPONENTS FOR BIOASSESSMENT AND
BIOMONITORING
Goal/Quality
Attribute
Accurate
Assessment of
Aquatic
Ecosystem
Integrity
Visibility
Ease of
Use and
Interpretation
Fish populations and individuals generally remain in the
same area during summer seasons.
Communities are persistent and usually recover rapidly from
natural disturbances. Comparable results can be expected
from an unperturbed-site at various times within a season.
Fish have larger home ranges and are less affected by
natural microhabitat differences than smaller organisms,
such as macroinvertebrates. This makes fish extremely
useful for assessing regional, macrohabitat, and mesohabitat
differences.
Most fish species have long life spans (3-10+ years) and can
reflect both long term and current water resource quality.
Fish continually inhabit the receiving water and reflect
the chemical, physical, and biological histories of the
water.
Fish represent a broad spectrum of community tolerances from
very sensitive to highly tolerant, and respond to chemical,
physical, and biological degradation in characteristics
response patterns.
Fish are a highly visible component of the aquatic
community, and so are of interest to the public.
Aquatic resource uses and regulatory language are generally
characterized in terms of fish (i.e., fishable and swimmable
goals of the Clean Water Act).
The sampling frequency for trend assessment is less than for
short-lived organisms.
The taxonomy of fishes is well established, allowing
professional biologists the ability to reduce laboratory
time by identifying many specimens in the field.
The distribution, life histories, and tolerances to
environmental stresses of most North American species are
well documented in the literature.
Adapted from Simon (1991).
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TABLE 2. FIVE MAJOR CLASSES OF ENVIRONMENTAL FACTORS WHICH INFLUENCE AND
DETERMINE THE BIOLOGICAL INTEGRITY OF SURFACE WATERS WITH SOME OF
THEIR IMPORTANT CHEMICAL, PHYSICAL, AND BIOLOGICAL COMPONENTS IN
LENTIC AND LOTIC SYSTEMS
1. ENERGY SOURCE
STREAMS, RIVERS
Nutrient cycling
Organic matter particle size
Primary productivity
Seasonal cycles
Solar radiation
2. WATER QUALITY/CHEMICAL VARIABLES
STREAMS, RIVERS
Adsorption
Alkal initv
DO
Hardness
Metals, other
Nutrients
Organics
pH
Solubility
Temperature
Turbidity
Water cycling
toxic substances
LAKES, RESERVOIRS, ESTUARIES, OCEANS
Nutrients cycling
Organic matter particle size
Primary productivity
Seasonal cycles
Solar radiation
LAKES, RESERVOIRS, ESTUARIES, OCEANS
Adsorption
Alkalinity
DO
Hardness
Metals, other
Nutrients
Organics
PH
Solubility
Temperature
Turbidity
Water cycling
toxic substances
3. HABITAT QUALITY
STREAMS, RIVERS
Bank stability
Canopy
Channel morphology {riffles, pools)
Current velocity
Gradient
Instream cover (woody debris)
Riparian vegetation
Siltation
Sinuosity
Substrate types
Width/depth
LAKES, RESERVOIRS, ESTUARIES, OCEANS
Bank stability
Shoreline vegetation
Substrate types
Siltation
Wave action
Width/depth
Inwater abiotic/biotic cover
Adapted from Karr (1987, 1991), Karr
1987), and USEPA (1990a; 1990b).
and Dudley (1981), Karr et al. (1986,
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TABLE 2. FIVE MAJOR CLASSES OF ENVIRONMENTAL FACTORS WHICH INFLUENCE AND
DETERMINE THE BIOLOGICAL INTEGRITY OF SURFACE WATERS WITH SOME OF
THEIR IMPORTANT CHEMICAL, PHYSICAL, AND BIOLOGICAL COMPONENTS IN
LENTIC AND LOTIC SYSTEMS (CONTINUED)
4. FLOW REGIME
STREAMS, RIVERS LAKES, RESERVOIRS, ESTUARIES, OCEANS
Ground water Ground water
High/low extremes High/low extremes
Land use Land use
Preci pi tati on/runoff Preci pi tati on/runoff
Water volume Water volume
5. BIOTIC ASSOCIATIONS
STREAMS, RIVERS LAKES, RESERVOIRS, ESTUARIES, OCEANS
Feeding Feeding
Competition Competition
Disease Disease
Parasitism Parasitism
Predation Predation
Reproduction Reproduction
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1.11 The utilization of biological components (structural and functional) to
evaluate the ambient aquatic community of our nations surface water has been
discussed and well documented in the literature. Some recent examples are
Crowder (1990), Downing et al. (1990), Fausch et al. (1990), Hunsaker and
Carpenter (1990), Karr et al. (1986), Karr, (1991), Ohio EPA (1987a, 1987b,
1989, 1990), Plafkin et al. (1989), Shuter (1990), Simon (1991), and USEPA
(1990a, 1990b). Structural components of fish communities include diversity,
taxa guilds, numbers, and biomass. Functional components of fish communities
include the feeding or trophic strategy, reproductive behavior and guild
classification, and environmental tolerance to perturbations.
1.12 The principal characteristics of interest in bioassessment studies of
fish populations include: (1) species richness (number of species)--presence
or absence; relative and absolute abundance of each species, (2) size
distribution, (3) habitat guilds--pelagic, littoral, and benthic species, (4)
trophic guilds--omnivores, piscivores, and invertivores, (5) growth rate, (6)
condition factor, (7) reproductive guilds, egg production and success, (8)
general tolerance guilds (indicator taxa)--intolerant, tolerant, and sensitive
species, (9) incidence of disease and parasitism (10) fish kills, (11)
palatability, and (11) fishability--catchability, desirability, and
sustainability. Observations of fish behavior can also be valuable in
detecting environmental problems, e.g., ventilation rates, position in the
current, and erratic movement. Fish may also be utilized for field and
laboratory bioassays (USEPA, 1991a, 1991b, 1992a, 1992b), for tissue analyses
to measure the concentrations of metals and pesticides (see Section 10,
Guidelines for Fish Sampling and Tissue Preparation for bioaccumulative
Contaminants) for histopathologic examination (Hinton and Lauren, 1990), and
biomarker studies (Adams, 1990a, 1990b; Anderson, 1990; Jimenez and Stegeman,
1990; Rice, 1990; Schreck, 1990; and Thomas, 1990).
1.13 Fisheries data are useful in enforcement cases and in long-term water
quality status and trends monitoring (Tebo, 1965; Ohio EPA, 1990; USEPA,
1991a). Before fishery surveys are initiated, a careful and exhaustive search
should be conducted for existing information on the fish populations or
communities in question. State and Federal fishery agencies and universities
may be potential sources of information. If data are not available and a
field stu'dy must be conducted, State and other Federal agencies may assist in
a survey and may provide needed expertise and specialized equipment for the
collection of specific, local fishes. A joint effort is usually more
economical and efficient and will promote continued cooperation between
agencies and parties involved.
1.14 Fisheries data may have limitations. Even if the species composition of
the fish in a specific area is known before and after the discharge of
pollutants, the significance of changes in the catch might not be
satisfactorily interpreted unless there are adequate data on spawning,
seasonal migration, temperature requirements and stream-flow responses,
feeding activities, diurnal movements, habitat preferences, and activity
patterns. Without adequate data, fish presence or absence cannot be directly
correlated with water quality. Furthermore, any existing data of known
quality on the water quality requirements of fish would be of value in
interpreting field data.
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1.15 Federal and state regulations usually require a fish collecting permit
because some species of fish are protected by law, and the collection of
others is regulated. The state fishery agencies must be contacted before fish
can be taken in a field study. Investigators should confirm that they have
complied with federal and state regulations before collecting samples of fish.
The state should be contacted prior to any fish study to ensure that
investigators comply with current regulations.
1.16 The design of fish studies should be based upon study goals and data
quality objectives (DQOs) (see Section 2, Quality Assurance and Quality
Control). To supplement the material contained in this manual, a number of
basic references should be reviewed by investigators involved in fish sampling
programs and studies. Useful references include Adams (1990), Angermeier et
al. (1991), APHA (1992), Bartell (1990), Edwards and Megrey (1989), Evans et
al. (1990), Everhart and Youngs (1981), Fausch et al. (1990), Gammon (1980),
Gammon et al. (1990), Hankin and Reeves (1988), Hellawell (1986), Herricks and
Schaeffer (1985), Hirsch et al. (1988), Hughes et al. (1986), Johnson and
Nielsen (1983), Karr (1981, 1987, 1990, 1991), Karr and Dionne, 1991, Karr and
Dudley (1981), Karr et al. (1983, 1986, 1987), Magnuson (1991), Manci (1989),
Mangel and Smith (1990), Minshall et al. (1989), Ohio EPA (1986, 1987a, 1987b,
1989, 1990), Omernik (1987), Platts et al. (1983), Robins et al. (1991),
Schreck and Moyle (1990), Tempieton (1984), Tonn (1990), USEPA (1988), USEPA
(1990a, 1990b), (USEPA, 1991c, 1991d, 1991e), Whittier and Paulsen (1992),
Wooten (1990), and Yoder (1991).
1.16.1 If fish data are to be useful, they must be acquired according to
standardized sampling methods and analyzed with appropriate statistical
methods. Two very important qualities of sampling data are accuracy and
precision. Accuracy refers to how well the sample represents the whole of the
study. In fishery studies, collecting accurate (or unbiased) data may be
difficult because studies are poorly designed. Precision refers to
repeatability of data. To supplement the statistics in this document,
investigators should consult the commonly cited statistical references
(Cochran, 1977; Conover, 1980; Green, 1979; Hicks, 1982; Snedecor and Cochran,
1981; Sokal and Rohlf, 1981; Zar, 1984).
1.17 Literature Cited
Adams, S.M. (ed.). 1990a. Biological indicators of stress in fish. American
Fisheries Symposium 8, American Fisheries Society, Bethesda, MD.
Adams, S.M. 1990b. Status and use of biological indicators for evaluating
the effects of stress on fish. In: Adams, S.M. (ed.). Biological
indicators of stress in fish. American Fisheries Society, Symposium 8,
American Fisheries Society, Bethesda, MD. pp. 1-8.
Anderson, D.P. 1990. Immunological indicators: Effects of environmental
stress on immune protection and disease outbreak. In: Adams, S.M,
Biological indicators of stress in fish. American Fisheries Society,
Symposium 8, American Fisheries Society, Bethesda, MD. pp. 38-50.
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Angermeier, P.L., R.J. Neves, and L.A. Nielsen. 1991. Assessing stream
values: Perspectives of aquatic resource professionals. North Amer. J.
Fisheries Management 11(1):1-10.
APHA. 1992. Standard methods for the examination of water and wastewater.
American Public Health Association, American Water Works Association,
and Water Pollution Control Federation, (18th ed.), Washington, DC.
Bartell, S.M. 1990. Ecosystem context for estimating stress-induced
reductions in fish populations. In: Adams, S.M. (ed.). Biological
indicators of stress fish. American Fisheries Symposium 8, American
Fisheries Society, Bethesda, MD. pp. 167-182.
Cochran, W.G. 1977. Sampling techniques. John Wiley and Sons, Inc., New
York, NY.
Conover, W.J. 1980. Practical nonparametic statistics. John Wiley, New
York, NY.
Crowder, L.B. 1990. Community ecology. In: Schreck, C.B. and P.B. Moyle
(eds.). Methods for fish biology. Amer. Fish. Soc., Bethesda, MD. pp.
609-632.
Downing, J.A., C. Plante, and S. Lalonde. 1990. Fish production correlated
with primary productivity, not the morphoedaphic index. Can. J. Fish.
Aquatic Sci. 47(10):1929-1936.
Edwards, E.F. and B.A. Megrey (eds.). 1989. Mathematical analysis of fish
stock dynamics. American Fisheries Symposium 6, American Fisheries
Society, Bethesda, MD.
Evans, D.O., G.J. Warren, V.W. Cairns. 1990. Assessment and management of
the fish community health in the Great Lakes: Synthesis and
recommendation. J. Great Lakes Res. 16(4):639-669.
Everhart, W.H. and W.D. Youngs. 1981. Principles of fishery science.
Cornell University Press, Ithaca, NY.
Fausch, K.D., J. Lyons, J.R. Karr, and P.L. Angermeier. 1990. Fish
communities as indicators of environmental degradation. In: Adams,
S.M. (ed.). Biological indicators of stress fish. American Fisheries
Symposium 8, American Fisheries Society, Bethesda, MD. pp. 123-144.
Gammon, J.R. 1980. The use of community parameters derived from
electrofishing catches of river fish as indicators of environmental
quality. In: Seminar on Water Quality Management Tradeoffs. EPA-
905/9-80-009, U.S. Environmental Protection Agency, Washington, DC.
Gammon, J.R., C.W. Gammon, and M.K. Schmid. 1990. Land use influence on fish
communities in central Indiana streams. In: W.S. Davis (ed.).
Proceedings of the 1990 Midwest Pollution Control biologists Meeting.
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EPA 905/9-90-995, U.S. Environmental Protection Agency, Chicago, IL.
pp. 111-120.
Green, R.H. 1979. Sampling design and statistical methods for environmental
biologists. John Wiley, New York, NY,
Hankin, D.G. and G, H. Reeves. 1988. Estimating total fish abundance and
total habitat area in small streams based on visual estimation methods.
Can. J. Fish. Aquat. Sci. 45(5):834-844.
Hellawell, J.M. 1986, Biological indicators of freshwater pollution and
environmental management. Elsevier Science Publishing, Co.,Inc.,
New York, NY
Herricks, E.E. and D.J. Schaeffer. 1985. Can we optimize biomonitoring?
Env. Mgmt. 9:487-492.
Hicks, C.R. 1982, Fundamental concepts in the design of experiments. Holt,
Rinehart, and Winston, New York, NY.
Hinton, D.E. and D.J, Lauren. 1990. Integrative histopathological
approaches to detecting effects of environmental stressors on fishes.
In: Adams, S.M. (ed.). Biological indicators of stress in fish.
American Fisheries Symposium 8, American Fisheries Society, Bethesda,
MD. pp. 51-66.
Hirsch, R.M., W.M. Alley, and W.G. Wilber, 1988. Concepts for a national
water-quality assessment program. U.S. Geological Survey Circular 1021,
Federal center, Denver, CO.
Hughes, R.M., D.P. Larsen, and J.M. Omernik. 1986. Regional reference sites:
a method for assessing stream pollution. Env. Mgmt. 10(5):629-635,
Jimenez, B.D. and J.J. Stegeman. 1990. Detoxication enzymes as indicators of
environmental stress on fish. In: Adams, S.M. (ed.). Biological
indicators of stress fish. American Fisheries Symposium 8, American
Fisheries Society, Bethesda, HD. pp. 67-79.
Johnson, D.L. and L.A. Nielsen. 1983. Sampling considerations. In;
Nielsen, L.A. and D.L. Johnson (eds.). Fisheries Techniques. American
Fisheries Society, Bethesda, MD. pp. 1-21.
Karr, J.R. 1981. Assessment of biotic integrity using fish communities.
Fisheries 6(6):21-27.
Karr, J.R. 1987. Biological monitoring and environmental assessment: a
conceptual framework. Environmental Management 11:249-256.
Karr, J.R. 1990a. Biological integrity and the good of environmental
legislation: Lessons for conservation biology. Conservation Biology
4:244-250.
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Karr, J.R. 1990b. Bioassessment and non-point source pollution: an overview.
Pages 4-1 to 4-18. In: Second National Symposium on Water Quality
Assessment. Meeting summary, October 16-19, 1989, Fort Collins,
Colorado, U.S. Environmental Protection Agency, Washington, DC.
Karr, J.R. 1991. Biological integrity: A long-neglected aspect of water
resource management. Ecological Applications 1:66-84.
Karr, J.R. and M. Dionne. 1991. Designing surveys to assess biological
integrity in lakes and reservoirs. In: Biological Criteria: Research
and Regulation. Proceedings of a symposium, pp. 62-72, EPA/440/5-91-
005. U.S. Environmental Protection Agency, Office of Water, Washington,
DC.
Karr, J.R. and D.R. Dudley. 1981. Ecological perspective on water quality
goals. Env. Mgmt. 5:55-68.
Karr, J.R., K.D. Fausch, P.L. Angermeier, P.R;. Yant, and I.J. Schlosser.
1986. Assessing biological integrity in running waters: a method and
its rationale. Special Publication 5. Illinois Natural History Survey,
Urbana, !L.
Karr, J.R., L.A. Toth, and 6.D. Garman. 1983. Habitat preservation for midwest
stream fishes: principles and guidelines. EPA-600/3-83-006. U.S.
Environmental Protection Agency, Corvallis, OR.
Karr, J.R., P.R. Yant, K.D. Fausch, and I.J. Schloser. 1987. Spatial and
temporal variability of the index of biotic integrity in three
midwestern streams. Trans. Amer. Fish. Soc. 116:1-11.
Magnuson, J.J. 1991. Fish and fisheries ecology. Ecol. Application 1(1):13-
26.
Manci, K.M. 1989. Riparian ecosystem creation and restoration: A literature
summary. Fish and Wildlife Service, U.S. Dept. Interior, Washington,
DC.
Mangel, M. and P.E. Smith. 1990. Presence-absence sampling for fisheries
management. Can. J. Fish. Aquat. Sci. 47:1875-1887.
McKenzie, D.H., D.E Hyatt, and V.J. McDonald (eds.). 1992. Ecological
Indicators. Proceedings of an International Symposium, Fort Lauderdale,
USA, October 16-19, 1990. Volume I and II. Elsevier Applied Science,
Elsevier Publishers, Ltd., London and New York.
Minshall, G.W., S.E. Jensen, and W.S. Platts. 1989. The ecology of stream
and riparian habitats of the Great Basin region: A community profile.
National Wetlands Research Center, U.S. Fish and Wildlife Service,
Slide!!, LA.
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Ohio EPA. 1986. The cost of biological field monitoring. Ohio Environmental
Protection Agency, Division of Water Qual. Monitoring and Assessment,
Evaluation and Standards Section, Columbus, OH.
Ohio EPA. 1987a. Biological Criteria for the protection of aquatic life:
Volume I: The role of biological data in water quality assessment.
Ohio Environmental Protection Agency, Division of Water Quality Planning
and Assessment, Ecological Assessment Section, Columbus, OH.
Ohio EPA. 1987b. Biological criteria for the protection of aquatic life:
Volume II. Users manual for biological field assessment of Ohio surface
waters. Ohio Environmental Protection Agency, Division of Water Quality
Planning and Assessment, Ecological Assessment Section, Columbus, OH.
Ohio EPA. 1989. Biological criteria for the protection of aquatic life:
Volume III. Standardized biological field sampling and laboratory
methods for assessing fish and macroinvertebrate communities. Ohio
Environmental Protection Agency, Division of Water Quality Planning and
Assessment, Ecological Assessment Section, Columbus, OH.
Ohio EPA. 1990. The use of biocriteria in the Ohio EPA surface water
monitoring and Assessment Program. Ohio Environmental Protection
Agency, Division of Water Quality Planning and Assessment, Ecological
Assessment Section, Columbus, OH.
Omernik, J.M. 1987. Ecoregions of the conterminous United States. Ann.
Assoc. Amer. Geogr. 77:117-125.
Plafkin, J.I,, M.T. Barbour, K.D. Porter, S.K. Gross, and R.M. Hughes. 1989.
Rapid bioassessment protocols for use in streams and rivers: benthic
macroinvertebrates and fish. EPA/440/4-89/001. U.S. Environmental
Protection Agency, Assessment and Watershed Protection Division,
Washington, DC.
Platts, W.S., W.F. Megahan, and G.W. Minshall. 1983. Methods for evaluating
streams, riparian, and biotic conditions. General Technical Report INT-
138, Intermountain Forest and Range Experiment Station, Forest Service,
U.S. Dept. Agriculture, Ogden, UT.
Rankin, E.T. 1989. The qualitative habitat evaluation index (QHEI):
rationale, methods, and application. Ohio Environmental Protection
Agency, Division Water Quality, Planning and Assessment, Ecological
Assessment Section, P.O. Box 1049, 1800 WaterMark Drive, Columbus, OH.
Rice, J.A. 1990. Bioenergetics modeling approaches to evaluation of stress
in fish. In: Adams, S.M. (ed.). Biological indicators of stress
fish. American Fisheries Symposium 8, American Fisheries Society,
Bethesda, MD. pp. 80-92.
Robin, C.R., C.E. Bond, J.R. Brooker, E.A. Lachner, R.N. Lea, and W.B. Scott.
1991. Common and scientific names of fishes from the United States and
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Canada. Fifth Edition. American Fisheries Society, Special Publication
20, American Fisheries Society, Bethesda, MD.
Schreck, C.B. 1990, Physiological, behavioral, and performance indicators of
stress. In: Adams S.M. (ed.). Biological indicators of stress fish.
American Fisheries Symposium 8, American Fisheries Society, Bethesda,
MD. pp. 29-37.
Schreck, C.B. and P.B. Moyle (eds.). 1990. Methods for fish biology.
American Fisheries Society, Bethesda, MD.
Shuter, B.J. 1990. Population-level indicators of stress. In: Adams, S.M.
(ed.). Biological indicators of stress in fish. American Fisheries
Symposium 8, American Fisheries Society, Bethesda, MD. pp. 145-166.
Simon, T.P. 1991. Development of index of biotic integrity expectations for
the ecoregions of Indiana. I. Central Corn Belt Plain. EPA-905/9-
91/025. U.S. Environmental Protection Agency, Environmental Science
Division, Monitoring and Quality Assurance Branch, Ambient Monitoring
Section, Chicago, IL.
Smith, P.M. 1971. Illinois streams: a classification based on their fishes
and an analysis of factors responsible for the disappearance of native
species. 111. Nat. Hist. Surv. Notes 76.
Snedecor, G.W. and W.6. Cochran. 1981. Statistical methods, Iowa State
University Press, Ames, IA.
Sokal, R.R. and F.J. Rohlf. 1981. Biometry, Freeman, San Francisco, CA.
Tebo, Jr., L.B. 1965. Fish population sampling studies at water pollution
surveillance system stations on the Ohio, Tennessee, Clinch, and
Cumberland Rivers. Applications and development Report No. 15, Div.
Water Supply and Pollution Control, U.S. Public Health Service,
Cincinnati, OH.
Templeton, R.G. 1984. Freshwater fisheries management. Fishing News Books,
Ltd., Farnham, Surrey, England, U.K.
Thomas. P, 1990, Molecular and biochemical responses of fish to stressors
and their potential use in environmental monitoring. In: Adams, S.M.
(ed.J. Biological indicators of stress fish. American Fisheries
Symposium 8, American Fisheries Society, Bethesda, MD. pp. 9-28.
Tonn, W.M. 1990. Climate change and fish communities: A conceptual
framework. Trans. Amer. Fish. Soc. 119:337-352.
USEPA. 1988. The lake and reservoir restoration guidance manual. EPA 440/5-
88-002. U.S. Environmental Protection Agency, Criteria and Standards
Division, Nonpoint Sources Branch, Washington, DC.
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USEPA. 1990a. Macroinvertebrate field and laboratory methods for evaluating
the biological integrity of surface waters. Donald J. Klemm, Philip A.
Lewis, Florence Fulk, and James M. Lazorchak. EPA/600/4-90/030.
U.S. Environmental Protection Agency, Environmental Monitoring Systems
Laboratory, Cincinnati, OH.
USEPA. 1990b. Biological Criteria. National Program guidance for Surface
Waters. EPA/440/5-90/004. U.S. Environmental Protection Agency, Office
of Water, Criteria and Standards Division, Office of Water Regulations
and Standards, Washington, DC.
USEPA. 1991a. Technical support document for water quality-based toxics
control. EPA/5052-90/001. U.S. Environmental Protection Agency, Office
of Water Enforcement and Permits and Office of Water Regulations and
Standards, Washington, DC.
USEPA. 1991b. Methods for measuring the acute toxicity of effluents and
receiving waters to freshwater and marine organisms. Cornelius I.
Weber (ed.). Fourth Edition. EPA/600/4-90/027. U.S. Environmental
Protection Agency, Monitoring Systems Laboratory, Cincinnati, OH.
USEPA. 1991c. Biological Criteria. State development and implementation
efforts. EPA-440/5-91-003. U.S. Environmental Protection Agency,
Office of Water, Washington, DC.
USEPA. 1991d. Biological criteria. Guide to technical literature. EPA-
440/5-91-004. U.S. Environmental Protection Agency, Office of Water,
Washington, DC.
USEPA. 1991e. Biological criteria: Research and regulation. EPA-440/5-91-
005. U.S. Environmental Protection Agency, Office of Water, Washington,
DC.
USEPA. 1992a. Short-term methods for estimating the chronic toxicity of
effluents and receiving waters to marine and estuarine organisms.
Donald J. Klemm and George E. Morrison (eds.). Second Edition.
EPA/600/4-91-021. U.S. Environmental Protection Agency, Environmental
Monitoring Systems Laboratory, Cincinnati, OH.
USEPA. 1992b. Short-term methods for estimating the chronic toxicity of
effluents and receiving waters to freshwater organisms. Philip A.
Lewis, Donald J. Klemm, and James M. Lazorchak (eds.). Third Edition.
EPA/600/4-91/022. U.S. Environmental Protection Agency, Environmental
Monitoring Systems Laboratory, Cincinnati, OH.
Whittier, T.R. and S.G. Paulsen 1992. The surface waters component of the
Environmental Monitoring and Assessment Program (EMAP): an overview. J.
Aquatic Ecosystem Health 1:119-126.
Wooten, R.J. 1990. The ecology of teleost fishes. Chapman and Hall Press,
New York, NY.
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Yoder, C.O. 1991. The integrated biosurvey as a tool for evaluation of
aquatic life use attainment and impairment in Ohio surface waters. In:
Biological Criteria: Research and Regulation. Proceedings of a
Symposium. EPA/440/5-91-005. U.S. Environmental Protection Agency,
Office of Water, Washington, DC. pp. 110-122.
Zar, J. H. 1984. Biostatistical analysis. Prentice-Hall, Inc., Englewood
Cliffs, NJ.
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SECTION 2
QUALITY ASSURANCE AND QUALITY CONTROL
2.1 Introduction
2.1.1 Fish studies, like macroinvertebrate studies (USEPA, 199Qa), require a
strong quality assurance (QA) program and effective quality control (QC)
procedures that encompass field and laboratory data collection activities.
The term "quality assurance" refers to an integrated system of activities
involving planning, quality control, quality assessment, reporting and quality
improvement to ensure that a product or service meets defined standards of
quality with a stated level of confidence. The term "quality control" refers
to the overall system of technical activities whose purpose is to measure and
control the quality of a product or service so that it meets the needs of
users. The aim is to provide quality that is satisfactory, adequate,
dependable, and economical (modified from USEPA, 1974; 1978).
2.1.2 Quality assurance programs have two primary functions in a
biomonitoring/bioassessment laboratory. First, the project or program should
define the data quality needed for the program's goals in terms of accuracy,
precision, representativeness, comparability, and completeness (see Subsection
2.6, Fish Collection). The second function is to provide information on the
success with which the measurement data meet these goals.
2.1.3 Quality assurance and quality control (QA/QC) must be a continuous
process in the biomonitoring/bioassessment program that includes all aspects
of the program, including field collection and preservation, habitat
assessment, sample processing, data analysis, and reporting. Otherwise, the
data generated may not be reliable and useful for decision making, and the
results will be of little use in assessing and establishing the conditions
(health, biological integrity, and quality of the water resources) of the
water body under study. Without an appropriate program of quality assurance
and quality control, data will be of unknown quality, limiting its
interpretation and usefulness. Quality must be assured before the results can
be accepted with any scientific studies. As described below, quality
assurance is accomplished through establishment of thorough investigator
training, protocols, guidelines, comprehensive field and laboratory data
documentation and management, verification of data reproducibility, and
instrument calibration.
2.1.4 To support the operation of a consistent plan, the persons responsible
for QA should consult the EPA Quality Assurance manual (USEPA, 1984a; 1984b;
1989; 1992b). All EPA QA programs are implemented and operated under the
authority of EPA Order 5360.1. USEPA (1984b) serves as guidance and describes
the policy, objectives, and responsibilities of all USEPA programs, regional
offices, and laboratories producing data for USEPA to institute a specific QA
program. Each office or laboratory that generates data under USEPA's QA/QC
program must implement, at a minimum, the prescribed procedures to ensure that
precision, accuracy, completeness, comparability, and representativeness of
data are known and documented.
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2.1.4.1 Information and discussion of statistical tools, data quality
objectives, comparison of good laboratory and field practices, and other
quality assurance considerations in the context of ecological research are
found in USEPA (1992b). Each agency should have a designated QA/QC officer
(or a person in charge of the program) responsible for reviewing project
plans, SOPs, etc. and auditing the program for improving performance, etc.
2.1.5 The Fish Bioassessment Protocols for Use In Streams and Rivers, Section
8, can be modified to achieve various data quality objectives. A different
habitat assessment approach, replicate sampling, more intensive sample
enumeration, or modified analytical metrics may be preferred by a particular
State over the approaches in this Section. Such refinements can be
accommodated, provided they are clearly documented in an USEPA approved QA
program and/or project plan.
2.1.6 Components of the QA program (Khalil and Tuckfield, 1992; USEPA, 1984a;
1984b; 1990a; 1991a; 1992a; 1992b) should include the following:
2.1.6.1 Approved methodology and documentation for the collection,
preservation, and analysis of data.
2.1.6.2 Documentation and manufacturer's instructions for sampling equipment,
flow measuring devices, and other measuring instruments such as pH, DO, and
conductivity meters.
2.1.6.3 Methods and documentation to assure that representative samples are
collected (See Subsection 2.2, Data Quality Objectives and Subsection 2.8,
Standard Operating Procedures).
2.1.6.4 Methods and documentation to assure the precision of sampling and
analysis procedures. Collecting precise fish data usually requires extensive
sampling as well as careful design.
2.1.6.5 Methods to assure accurate and timely recording, storage, and
retrieval of data.
2.1.6.6 Documentation to assure sample evaluation, statistical evaluation,
and performance evaluation of laboratory procedures.
2.2 Data Quality Objectives
2.2.1 A full assessment of the data quality needed to meet the study
objectives should be made prior to preparation and implementation of the QA
plan. Data quality is a measure or description of the completeness, type, and
amount of error associated with a data set. Determination of data quality is
accomplished through the development of data quality objectives (DQOs), which
are statements of the level of uncertainty a decision-maker is willing to
accept or the quality of the data needed to support a specific environmental
decision or action and the rationale behind those statements and levels of
data quality. Both qualitative and quantitative descriptors of data quality
must be considered to determine whether data are appropriate or adequate for a
particular application. However, DQOs are target values and not necessarily
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criteria for the acceptance or rejection of data (Table 1). Table 1 is a
summary listing QA objectives for precision and completeness. Data quality
requirements should be based on prior knowledge of the sampling procedures or
measurements system by use of replicate (duplicate) analyses, reference
conditions (site-specific or ecoregional), or requirements of the specific
project (USEPA, 1989).
2.2.2 Data quality objectives are developed in three stages. During the
first stage, the decision-maker determines what information is needed, reasons
for the need, how the information will be used, and specifies time and
resource constraints. The second stage involves the technical staff and the
decision-maker interacting to establish a detailed and clarified specification
of the problem, how the information will be used, any constraints imposed on
the data collection, and what limitations of the information will be
acceptable. The third stage involves the examination of the possible
approaches to collection and analysis of the data and a determination of the
quality of the data that can be expected to result from each approach. The
best approach is selected based upon the criteria agreed upon in the second
stage. It may be necessary to modify the objectives of the study during the
development of the DQOs. Details for developing DQOs are described in USEPA
(1986; 1989). These documents are available from the Quality Assurance
Management Staff, Office of Research and Development, Washington, DC 20460 and
the Center For Environment Research Information (CERI), U.S. Environmental
Protection Agency, Cincinnati, OH 45268. The CERI information and document
ordering phone number is (513) 569-7562. Johnson and Nielsen (1983), Ohio EPA
(1989), and Simon (1991) discuss sampling considerations for collecting fish
data.
2.2,3 After the DQOs are established, the detailed project QA plan should be
finalized stating specific quantitative and qualitative data quality goals and
QC procedures that will be used to control and characterize error (USEPA,
1980; 1989; 1992b). These goals, based on the DQOs, will be the criteria for
measuring the success of the QA program.
2.2.4 The Quality Assurance Management Staff, Office of Modeling, Monitoring
Systems, and Quality Assurance, is responsible for providing general guidance
for the inclusion of DQOs in quality assurance program and project plans, and
for providing guidance to the regions on the application of the DQOs
development process. The EPA regional offices are responsible for ensuring
that state QA programs and project plans are in conformance with grant
requirements specified in 40 CFR Part 30, and for assisting the states in
developing DQOs requirements and Quality Assurance Program Plans (QAPP) that
meet state needs (USEPA, 1989).
2.2.5 Regional and state laboratories or monitoring personnel in need of
specific guidance in preparing Quality Assurance Project Plans or development
of DQOs for bioassessment projects can contact personnel of the Bioassessment
and Ecotoxicology Branch in the Ecological Monitoring Research Division,
Environmental Monitoring Systems Laboratory-Cincinnati, OH for assistance
((513) 533-8114, FAX (513) 533-8181).
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TABLE 1. EXAMPLE OF SUMMARY TABLE FOR DATA QUALITY REQUIREMENTS1
Measurement Precision
Parameter Reference (RPD2, RSD3) Completeness (%)
Benthos Plafkin et al. (1989)
No. Individuals 50 95
No. Taxa 15 95
Fish Karr et al. (1986)
No. Individuals 25 95
No. Species 15 95
Dissolved Oxygen (mg/L) ASTM (1992) 5 90
Water Temperature °C ASTM (1992) 5 90
1From USEPA (1992b).
2RPD = Relative percent difference.
3RSD = Relative standard deviation.
2.3 Facilities And Equipment
2.3.1 Laboratory, field facilities, and equipment must be in place and
operating consistently with their designed purposes so that quality
environmental data may be generated and processed in an efficient and cost-
effective manner. Suitability of the facilities for the execution of both the
technical and QA aspects of the study should be assessed prior to initiation
of the study. Adequate environmental controls (space, lighting, temperature,
noise levels, and humidity) should be provided. Satisfactory safety and
health maintenance features must also be provided (see Section 3, Safety and
Health).
2.3.2 Equipment (boats, sampling gear, etc.) and supplies necessary to
adequately collect, preserve and process fish and other biological samples
must be available and in good operating condition. See Section 4, Sample
Collection for Analysis of the Structure and Function of Fish Communities,
Table 3, General Checklist Of Fish Field Equipment And Supplies.
2.3.3 To ensure data of consistently high quality, a plan of routine
inspection and preventive maintenance should be developed for all facilities
and equipment. All inspections, calibrations, and maintenance must be
documented in individual bound notebooks. This documentation should include
detailed descriptions of all calibrations performed, adjustments made, and
parts replaced, and each entry should be signed and dated.
18
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2.4 Calibration, Documentation, and Record Keeping
2.4.1 Quality assurance plans should contain mechanisms for demonstrating the
reproducibility of each measuring process. Regular calibration of
instruments, proper documentation, and permanent record keeping are essential
aspects of such plans.
2.4.2 Each measuring device (pH and DO meters, etc.) must be calibrated
before each use according to the manufacturer's instructions, and routine
checks using National Institute of Standards and Technology standards, or
other standards of known accuracy, should be made to demonstrate that
variables are within predetermined acceptance limits. Permanent records
giving dates and details of these calibrations and checks must be kept.
Documentation is necessary to identify each specific measuring device, where
and when it is used, what maintenance was performed, and the dates and steps
used in instrument calibration. All samples collected and field data sheets
should also be assigned a unique identification number and label. Data should
be documented to allow complete reconstruction, from initial field record
through data storage system retrieval.
2.4.3 Sample tracking is important, but whenever samples are collected to be
used as evidence in a court of law, it is imperative that laboratories and
field operations follow written chain-of-custody procedures for collecting,
transferring, storing, analyzing, and disposing of the samples. The primary
objective of chain-of-custody procedures is to create a written record
(Figures 1 and 2) can be used to trace the possession of the sample from the
moment of collection through the introduction of the analytical data into
evidence. Explicit procedures must be followed to maintain the documentation
necessary to satisfy legal requirements. All survey participants should
receive a copy of the study plan and be knowledgeable of its contents prior to
implementing the field work. A presurvey briefing should be held to
reappraise all participants of the survey objectives and chain-of-custody
procedures. After all chain-of-custody samples are collected, a debriefing
should be held in the field to check adherence to chain-of-custody procedures.
Chain-of-custody procedures are discussed in four USEPA manuals (USEPA, 1974;
1990b; 1991a; 1992b).
2.4.4 Field and laboratory personnel should keep complete, permanent records
of all conditions and activities that apply to each individually numbered
sample sufficient to satisfy legal requirements for any potential enforcement
or judicial proceedings. The field data sheets and sample tags (see Section
4, Sample Collection for Analysis of the Structure and Function of Fish
Communities; Section 5, Fish Specimen Processing; Section 8, Fish
Bioassessment Protocols For Use In Streams and Rivers) should be filled out as
completely and as accurately as possible to provide a record in support of the
survey and analysis conclusion. Abbreviations commonly used in documentation
(e.g., scientific names) should be standardized to decrease data manipulation
error. Field and laboratory data sheets and final reports should be filed.
All field and laboratory data sheets should be dated and signed by the sampler
and analyst, respectively. Notebooks, data sheets, and all other records that
may be needed to document the integrity of the data should be permanently
filed in a secure fireproof location.
19
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Project No.
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Figure 1. Example of sample identification tag.
USEPA (1991a).
From USEPA (1990b) and
2.5 Habitat Assessment
2.5.1 Because the habitat characterization procedures (see Section 4, Sample
Collection for Analysis of the Structure and Function of Fish Communities and
Section 8, Fish Bioassessment Protocols for Use in Streams and Rivers) are
primarily a qualitative evaluation, final conclusions are potentially subject
to variability among investigators. This limitation can be minimized however,
by ensuring that each investigator is appropriately trained in the habitat
evaluation techniques and periodic cross-checks are conducted among
investigators to promote consistency. Also, bioassessment laboratories should
institute one or two day training courses on habitat characterization and
evaluation followed by periodic refresher training. For additional
information and discussion on habitat evaluation and a Qualitative Habitat
Evaluation Index (QHEI), see Barbour and Stribling (1991), Plafkin et al.,
(1989), Ohio EPA (1989), Rankin (1989), and USEPA (1990a; 1991b) for
additional information and discussion on habitat evaluation and a Qualitative
Habitat Evaluation Index (QHEI), regarding rationale, methods, and application
for fish bioassessment. Also, see Section 4, Sample Collection for Analysis
of the Structure and Function of Fish Communities, Subsection 4.1.5, Habitat
Evaluation and Section 8, Fish Bioassessment Protocols For Use In Streams and
Rivers, Subsection 8.13.3, Habitat Quality and Assessment.
20
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STATION NO.
PROJECT NO.
PROJECT LEADER
PROJECT NAME/LOCATION
SAMPLE TYPES
1. SURFACE WATER
2. GROUND WATER
3. POTABLE WATER
«. WASTEWATER
5. UEACHATf
11. OTHER
6. SOIL/SEDIMENT
7. SLUDGE
8. WASTE
9. UK
10. FISH
19
DATE
JtML
SAMPLERS (SIGN)
STATION LOCATION/DESCRIPTION
REMARKS
CIRCLE/ADD
parameter:
desired.
List no. of
containers
submitted
ANALYSES
TAG NO./REMARKS
LAB
USE
ONLY
RELINQUISHED BY:
(PRINT)
(3CN)
DATE/TIME
RECEIVED BY:
(PRINT)
RELINQUISHED BY:
(PRINT)
(SIM)
DATE/TIME
RECEIVED BY;
(PRINT)
(3CN) .
RELINQUISHED BY:
(PRINT)
DATE/TIME
RECEIVED 8Y:
(PRINT)
RELINQUISHED BY:
(PRINT)
DATE/TIME
RECEIVED BY:
(PRINT)
(SOU)
(SIGN)
(3BN)
(3CN)
DISTRIBUTION: VKilte and Pink copies accompany sample shipment to laboratory; Pink copy retained by laboratory,
WiHe copy is returned to samplers; Yellow copy retained by samplers.
•U.S. CPO: 1989-732-186
4-20043
(10/89)
Figure 2. Example of a chain-of-custody record form. Modified from USEPA (1990b), USEPA (1991a), and
USEPA, Region 4.
-------
2.6 Fish Collection
2.6.1 Ensuring that fish field survey data are representative of the fish
assemblage at a particular site requires careful regional analysis and station
evaluation. Data comparability is maintained by using similar collection
methods and sampling effort in waterbodies (lakes, reservoirs, estuaries,
wetlands, streams, rivers, etc.) of similar size . Also, where possible,
major habitats in streams (riffle, run, pool) are sampled at each site, and
the proportion of each habitat type sampled should be noted.
2.6.2 Precision, accuracy, and completeness should be evaluated in pilot
studies along with sampling methods and site size. Variability among
replicates from the same site or similar sites should not produce differences
exceeding 10 percent at minimally impacted sites and 15 percent at highly
impacted sites (Plafkin et al., 1989). Index of Biological Integrity (IBI)
differences at the same site should not exceed 4 (Karr et al., 1986).
2.6.3 Data reproducibility may be ensured by having a variety of
investigators periodically resample well characterized sites. Investigator
precision and accuracy for use of the Index of Biological Integrity (IBI) and
the Index of well-being (Iwb) may be determined by having investigators
evaluate a standard series of data sets or preserved field collections.
2.6.4 Taxonomists, fishery staff, and aquatic biologists should be capable of
identifying fish to the lowest possible level (species, subspecies) and
should have at their disposal adequate taxonomic references to perform the
level of identification required. See Section 12, Fisheries Bibliography, for
a list of selected taxonomic references. Fishery and aquatic biologists
should check this list and obtain those references that will be needed for the
identification of specimens.
2.6.5 Field identifications are acceptable, but laboratory voucher specimens
are always required for new locality records, new species, and any specimens
that cannot be identified in the field. All specimens should be retained for
laboratory examination if there are any doubts about the correct
identification. Biomonitoring laboratories that do not identify fish and
other taxa on a regular basis or that have difficulty identifying organisms
should have representative specimens of all taxa verified by a specialist who
is a recognized authority in that particular taxonomic group. These specimens
must be properly labeled as reference or voucher specimens, including the name
of the verifying authority, permanently preserved, and stored in the
laboratory, or voucher specimens should be offered to regional and state
natural history museums for future reference.
2.6.6 Quality control of taxonomic identifications is accomplished by a
second qualified individual.
2.7 Qualifications and Training
2.7.1 All personnel need to have adequate education, training, and experience
in the areas of their technical expertise, responsibilities, and in quality
22
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assurance (QA). Because no formal academic programs in research QA exist,
most QA experience must be acquired through on-the-job training.
2.7.2 At least one professional biologist with training and experience in
fish sampling methods and fish identification should be involved directly in
the field work or should be involved for at least the first two weeks of the
field sampling season (and thereafter if necessary), instructing other less
qualified staff in all aspects of the field sampling as well as the laboratory
analysis of the samples to ensure data quality. Additionally, the
investigators should be familiar with the objectives of each site
investigation. Periodic conferences with the sampling crew to assure the
sampling effort is being conducted in accordance with the standard operating
procedures are also advisable. Statistical expertise should be readily
available and consulted during every phase of the project.
2.7.3 Management should periodically assess the training needs of all
personnel engaged in QA, and recommend and support their participation in
appropriate and relevant seminars, training courses, and professional
meetings.
2.7.4 Project personnel should have on file an up-to-date resume for each
person who is responsible for the collection, analysis, evaluation and
reporting of biological data.
2.8 Standard Operating Procedures (SOPs)
2.8.1 Each laboratory should define the precise methods to be used during
each step of the collection, analysis, and data evaluation process. These
written procedures become the standard operating procedures (SOPs) describing
the operation of the laboratory (USEPA, 1991a). Standard operating procedures
for a fish laboratory should describe in stepwise fashion, easily understood
by the potential user, at least the following:
1. Sampling methodology, including maintenance of electrofishing gear and
seines
2. Replication (duplication)
3. Habitat assessment methodology
4. Sampling site and station selections (including reference sites)
5. Details of preservation and labeling of the samples
6. Use of taxonomic keys
7. Use and calibration of measuring instruments (e.g., DO, pH, and
conductivity meters, etc.) and QC requirements
8. Sample chain-of-custody and handling procedures
9. Data analysis, evaluation, and handling
23
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2.8.2 The SOPs must include a listing of the taxonomic keys and references
that should be used for each level of identification required and for each
taxonomic group. Field experience and taxonomic expertise requirements of
personnel for the particular level of bioassessment performed must be defined
in the preparation of DQOs. It should also provide an outline of the steps to
be taken to assure the quality of the data.
2.8.3 The SOPs must stress the need for the traceability of the fish samples.
At a minimum it should specify that the fish sample be assigned a unique
identification number and be properly labeled with the sample number, sampling
location, date, and name of the collector (see Section 5, Specimen Processing
Techniques for an example of sample tags). It should describe procedures to
ensure that each sample collected, as accurately and precisely as possible,
represents the fish community sampled.
2.8.4 The SOPs should be approved by the proper authority and must be easily
accessible to all appropriate personnel for referral.
2.8.5 The laboratory SOPs must be followed as closely as possible. Any
deviations should be documented as to the reason for the deviation and any
possible effect the deviation might have on the resulting data.
2.8.6 Field validation, conducted at a frequency to be determined by each
agency, should involve two procedures: (1) collection of replicate samples at
various stations to check on the precision and accuracy of the collection
effort, and (2) repeat field collections and analyses performed by separate
field crews to provide support for the bioassessment. In addition, field
crews should occasionally alternate personnel with the same field training to
maintain objectivity in the bioassessment study.
2.9 Literature Cited
ASTM. 1992. Standard test methods for dissolved oxygen in water. D 888-87.
Annual book ASTM standards: Water and environmental technology.
American Society of Testing and Materials, Philadelphia, PA. pp. 522-
533.
Barbour, M.T. and J.B. Stribling. 1991. Use of habitat assessment in
evaluating the biological integrity of stream communities. In:
Biological Criteria: Research and Regulation, 1991. EPA-440/5-91-005.
U.S. Environmental Protection Agency, Office of Water, Washington, DC.
pp. 25-38.
Johnson, D.L. and L.A. Nielsen. 1983. Sampling considerations. In:
Nielsen, L.A. and D.L. Johnson (eds.). Fisheries techniques. American
Fisheries Society, Bethesda, MD. pp. 1-21.
Karr, J. R., D. D. Fausch, P. L. Angermeier, P. R. Yant, and I. J. Schlosser.
1986. Assessing biological integrity in running waters: A method and
its rationale. Special Publication 5. Illinois Natural History Survey.
24
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Khalil, M.M. and R.C. Tuckfield. 1992. A quality assessment program for
monitoring laboratory performance. American Environmental Laboratory
4/92:8-14.
Ohio EPA. 1989. Biological criteria for the protection of aquatic life III:
Standardized biological field sampling and laboratory methods for
assessing fish and macroinvertebrate communities. Ohio Environmental
Protection Agency, Division of Water Quality Monitoring and Assessment,
Ecological Assessment Section, Columbus, OH.
Plafkin, J.L., M.T. Barbour, K.D. Porter, S.K. Gross, and R.M. Hughes. 1989.
Rapid bioassessment protocols for use in streams and rivers. Benthic
macroinvertebrates and fish. EPA/440-4-89/001. U.S. Environmental
Protection Agency, Office of Water, Assessment and Watershed Protection
Division, Washington, DC.
Rankin, E.T. 1989. The qualitative habitat evaluation index (QHEI):
rationale, methods, and application. Ohio Environmental Protection
Agency, Division of Water Quality Monitoring and Assessment, P.O. Box
1049, 1800 WaterMark Drive, Columbus, OH.
Simon, T.P. 1991. Development of index of biotic integrity expectations for
the ecoregions of Indiana. I. Central corn belt plain. Environmental
Science Division, Monitoring and Quality Assurance Branch, Ambient
Monitoring Section, U.S. Environmental Protection Agency, Chicago, IL.
USEPA. 1974. Model state monitoring program. EPA-440/9-74-002. U.S.
Environmental Protection Agency, Office of Water and Hazardous Materials,
Monitoring and Data Support Division, Washington, DC.
USEPA. 1978. Quality Assurance Newsletter. U.S. Environmental Protection
Agency, Environmental Monitoring and Support Laboratory - Cincinnati, OH.
USEPA. 1980. Guidelines and specifications for preparing quality assurance
project plans. Report No. QAMS-005/80. U.S. Environmental Protection
Agency, Office of Monitoring and Quality Assurance, Office of Research
and Development, Washington, DC.
USEPA. 1984a. Guidance for preparation of combined work/quality assurance
project plans for environmental monitoring. Report No. OWRS QA-1, U.S.
Environmental Protection Agency, Washington, DC.
USEPA. 1984b. Policy and program requirements to implement the quality
assurance program. EPA Order 5360.1, U.S. Environmental Protection
Agency, Washington, DC.
USEPA. 1986. Development of data quality objectives. Descriptions of stages
I and II. Prepared by the Quality Assurance Management Staff. U.S.
Environmental Protection Agency, Office of Research and Development,
Washington, DC.
25
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USEPA. 1989, Preparing perfect project plans. A pocket guide for the
preparation of quality assurance project plans, EPA/6009-89/087, U.S.
Environmental Protection Agency, Office of Research and Development, Risk
Reduction Engineering Laboratory, Cincinnati, Oh,
USEPA. 1990a. Macroinvertebrate field and laboratory methods for evaluating
the biological integrity of surface waters, Klemm, D.J., P.A. Lewis, F.
Fulk, and J.M. Lazorchak. EPA/600/4-90/003. U.S. Environmental
Protection Agency, Environmental Monitoring Systems Laboratory,
Cincinnati, OH.
USEPA, 1990b. Manual for the certification of laboratories analyzing drinking
water: Criteria and procedures - Quality assurance, EPA-570/9-90/Q08.
U.S. Environmental Protection Agency, Office of Water, Washington, DC.
USEPA. 1991a. Manual for the evaluation of laboratories performing aquatic
toxicity tests, Klemm, D.J., L.B. Lobring, and W.H. Horning, II,
EPA/600/4-90/031. U.S. Environmental Protection Agency, Environmental
Monitoring Systems Laboratory, Cincinnati, OH.
USEPA. 1991b. Biological Criteria, Guide to technical literature. EPA-
440/5-91-004. U.S. Environmental Protection Agency, Office of Water,
Washington, DC.
USEPA. 1992a. Fourth annual ecological quality assurance workshop.
EPA/600/R-92/097. U.S. Environmental Protection Agency, Office Research
and Development, Washington, DC.
USEPA. 1992b (Draft). Generic quality assurance project plan. Guidance for
bioassessment/biomonitoring programs. James M. Lazorchak and Donald J.
Klemm (eds.). U.S. Environmental Protection Agency, Environmental
Monitoring Systems Laboratory, Cincinnati, OH.
26
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SECTION 3
SAFETY AND HEALTH
3.1 Introduction
3.1.1 Collection and analysis of fish samples can involve significant risks
to personal safety and health (drowning, electrical shock, pathogens, etc.).
While safety is often not considered an integral part of a fish sampling
routine, the biologist must be aware of unsafe working conditions, hazards
connected with the operation of sampling gear, boats, and other risks (Berry
et al., 1983). Management should assign health and safety responsibilities
and establish a program for training in safety, accident reporting, and
medical and first aid treatment. The laboratory safety document and standard
operating procedures (SOPs) containing necessary and specific safety
precautions should be available to all persons involved in fish sample
collecting and processing. Field and laboratory safety requirements for
biomonitoring laboratories are found also in USEPA (1986) and Ohio EPA (1990).
3.2 General Precautions
3.2.1 Good housekeeping practice should be followed both in the field and in
the laboratory. These practices should be aimed at protecting the staff from
physical injury, preventing or reducing exposure to hazardous or toxic
substances, avoiding interferences with laboratory operations, and producing
valid data.
3.2.2 Field personnel and sampling crew must have mandatory training in Red
Cross first aid, cardiopulmonary resuscitation (CPR), boating and water
safety, field survey safety (weather conditions, personal safety, and vehicle
safety), presurvey safety requirements (equipment design, equipment
maintenance, reconnaissance of survey area), and electrofishing safety (Ohio
EPA, 1990). It is the responsibility of the group safety officer or field
sampling leader to ensure that the necessary safety courses are taken by all
field personnel and that all safety policies and procedures are followed.
3.2.3 Operation of fish sampling devices involves potential hazards that must
be addressed by the individuals using the equipment. Electrofishing equipment
should be operated carefully. Electrofishing should always be done with at
least three individuals, and all safety procedures must be followed. Persons
using these devices should become familiar with the hazards involved and
establish appropriate safety practices prior to using them (Reynolds, 1983;
Ohio EPA, 1990). Note: Individuals involved in electrofishing must be
trained by a person experienced in this method or by attending a certified
electrofishing training course (See Section 4, Sample Collection for Analysis
of the Structure and Function of Fish Communities, Subsection 4.3
Electrofishing and Ohio EPA, 1990).
3.2.4 Field personnel should be able to swim. Waders should always be worn
with a belt to prevent them from filling with water in case of a fall. The
27
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use of a life jacket is advisable at dangerous wading stations if one is not a
strong swimmer because of the possibility of sliding into deep water.
3.2.5 Individuals sampling with scuba gear must be certified. The hazards of
sampling with scuba gear are sufficiently great that certification is
mandatory.
3.2.6 Many hazards lie out of sight in the bottoms of lakes, rivers and
streams. Broken glass or sharp pieces of metal embedded in the substrate can
cause serious injury if care is not exercised when walking or working with the
hands in such environments. Infectious agents and toxic substances that can
be absorbed through the skin or inhaled may also be present in the water or
sediment.
3.2.7 Personnel must consider and prepare for hazards associated with the
operation of motor vehicles, boats, winches, tools, and other incidental
equipment. Boat operators should be familiar with U.S. Coast Guard rules and
regulations for safe boating contained in a pamphlet, "Federal Requirements
for Recreational Boats," available from your local U.S. Coast Guard Director
or Auxiliary, or State Boating Official (U.S. Coast Guard, 1987).
3.2.8 Prior to a sampling trip, personnel should determine that all necessary
equipment is in safe working condition and that the operators are properly
trained to use the equipment.
3.2.9 Safety equipment and first aid supplies must be available in the
laboratory and in the field at all times. All motor vehicles and boats with
motors must have fire extinguishers, boat horns, cushions, and flares or
communication devices.
3.3 Safety Equipment and Facilities
3.3.1 Necessary and appropriate safety apparel such as waders, lab coats,
gloves, safety glasses, and hard hats must be available and used in accordance
with the project safety plan.
3.3.2 First aid kits, fire extinguishers and blankets, safety showers, and
emergency spill kits must be readily available in the laboratory at all times.
3.3.3 A properly installed and operating hood must be provided in the
laboratory for use when working with carcinogenic chemicals (e.g.,
formaldehyde) that may produce dangerous fumes.
3.3.4 Communication equipment and posted emergency numbers must be available
to field personnel and those working in mobile labs in remote areas for use in
case of an emergency.
3.3.5 Facilities and supplies must be available for cleaning of exposed body
parts that may have been contaminated by pollutants in the water. Soap and an
adequate supply of clean water or ethyl alcohol, or equivalent, should be
suitable for this purpose.
28
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3.4 Field and Laboratory Operations
3,4,1 At least two persons (three persons for electrofishing) must be present
during all sample collection activities.
3.4.2 All surface waters should be considered potential health hazards due to
toxic substances or pathogens and exposure to them should be minimized as much
as possible. Exposed body parts should be cleaned immediately after contact
with these waters.
3.4.3 All electrical equipment must bear the approval of Underwriters
Laboratories and must be properly grounded to protect against electric shock.
3.4.4 Use a winch for retrieving large fish nets, trawls, etc., for samples
collected with heavy sampling devices, and use care in lifting heavy items to
prevent back injury.
3.4.5 Persons working in areas where poisonous snakes may be encountered must
check with the local Drug and Poison Control Center for recommendations on
what should be done in case of a bite from a poisonous snake. If local advice
is not available and medical assistance is more than an hour away, carry a
snake bite kit and be familiar with its use. Any person allergic to bee
stings or other insect bites must take proper precautions and have any needed
medications handy.
3.4,6 Personnel participating in field activities on a regular or infrequent
basis should be in sound physical condition and have a physical exam annually
or in accordance with Regional or State Safety requirements.
3.4.7 All field personnel should be familiar with the symptoms of hypothermia
and know what to do in case symptoms occur. Hypothermia can kill a person at
temperatures much above freezing (up to 10°C or 50°F) if he or she is exposed
to wind or becomes wet.
3.5 Disease Prevention
3.5.1 Unknown pollutants and pathogens in surface waters and sediments should
be considered potential health hazards and exposure to them kept to a minimum.
3.5.2 Personnel who may be exposed to water known or suspected to contain
human or animal wastes that carry causative agents or pathogens must be
immunized against tetanus, hepatitis, typhoid fever, and polio. Field
personnel should also protect themselves against the bite of deer or wood
ticks because of the potential risk of acquiring pathogens that cause Rocky
Mountain spotted fever and Lyme disease.
3.6 Literature Cited
Berry, C.R. Jr., W.T. Helm, and J.M. Neuhold. 1983. Safety in fishery
field work. In: Nielsen, L.A., and D.L. Johnson (sus.). Fisheries
Techniques, American Fisheries Society, Bethesda, MD. pp. 43-60.
29
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Ohio EPA. 1990. Ohio EPA Fish evaluation group safety manual. Ohio
Environmental Protection Agency, Ecological Assessment Section, Division
of Water Quality Planning and Assessment, Columbus, OH.
Reynolds, J.B. 1983. Electrofishing. In: L.A. Nielsen and D.L. Johnson
(eds.). Fisheries Techniques. Amer. Fish. Soc., Bethesda, MD. pp.
147-163.
U.S. Coast Guard. 1987. Federal requirements for recreational boats. U.S.
Department of Transportation, United States Coast Guard, Washington, DC.
USEPA. 1986. Occupational health and safety manual. Office of Planning
and Management, U.S. Environmental Protection Agency, Washington, DC.
30
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SECTION 4
SAMPLE COLLECTION FOR ANALYSIS OF THE STRUCTURE AND FUNCTION
OF FISH COMMUNITIES
4.1 General Considerations
4.1.1 A variety of methods, techniques, and equipment exist to sample fish
populations and communities in lentic and lotic habitats. In addition, many
procedures are available to analyze the fish data collected. Each technique
has different assumptions, advantages and disadvantages. It is important to
understand the attributes and characteristics of sampling equipment and
techniques used in fish biosssessment so that valid conclusions can be drawn
from the data. Sampling considerations and design (APHA, 1992; Lagler, 1956;
Johnson and Nielson, 1983; Schreck and Moyle, 1990; Section 2, Quality
Assurance and Quality Control) are important because aquatic biologists or
fisheries scientists spend a major part of their time collecting data and the
study results are determined by use of the data with a variety of techniques
and equipment for an assortment of studies. Since fish populations are
usually nonrandomly distributed and clumped in response to many habitat
variables (Allen et al., 1992; Hendricks et al., 1980), the choice of sampling
methods and equipment, the habitat and time of sampling, and frequency of
sampling will depend on the data quality objectives of the study. For
practical considerations, it is often easier to sample at certain places or
time of the year (e.g., shallow water areas or during low flow). Therefore,
all sampling gear is generally considered selective in sample collection to
some degree (Everhart et al., 1975; Gulland, 1980; Henderson, 1980; Lagler,
1956, 1978; Ricker, 1971; Schnick and Moyle, 1990; Yen, 1977; Zippin, 1956,
1958). Some procedures to reduce sampling bias through better sampling design
are found in Armour et al. (1983), Cyr et al. (1992), Gulland (1980); Johnson
and Nielsen (1983). The accurate and efficient collection of data can mean
the difference between a successful management and research effort and a study
that might end with inconclusive or inappropriate data.
4.1.2 In all bioassessment studies key physical, chemical, and biological
indicators or parameters to be monitored should be selected carefully for the
most direct cause and effect relationships. Some important indicators or
parameters of biological integrity for consideration are found in Table 1.
For a discussion of these variables and others, see Armour et al. (1983),
Lagler (1956, 1978), Orth (1983), Plafkin et al. (1989), Rankin (1989), Ohio
EPA (1987a, 1987b, 1989), and Section 8, Fish Bioassessment Protocols for Use
In Streams and rivers, Subsection 8.13 Habitat Assessment and Physical/
Chemical Parameters, and references in Section 12, Fisheries Bibliography.
4.1.3 Table 2 is a general list of equipment and supplies needed for the
collection of fish samples and biosurvey. The data quality objectives (DQOs),
standard operating procedures (SOPs), sampling and analysis methods should
determine the type of gear and supplies needed.
4.1.4 Figure 1, A-C are examples of fish field data sheets that can be
adapted for field collections. Table 3 contains codes that can be used to
31
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TABLE 1. GENERAL INDICATORS OF BIOLOGICAL/ECOLOGICAL INTEGRITY FOR FISH
Lakes and Reservoirs
Streams and Rivers
Structure and Function Components of Fish Populations and Communities
Species composition
Relative abundance
Biomass
Lengths
Weights
Age and growth
Condition factor
Population numbers
Fecundity
Indices IBI
Health/Condition profile
Gross pathology, parasitism,
disease incidence
State fish kills
Ice cover period
Pollution indices
Ichthyoplankton index
Species composition
Relative abundance
Biomass
Lengths
Weights
Age and growth
Condition factor
Population numbers
Fecundity
Indices IBI/Iwb
Health/condition profile
Gross pathology, parasitism,
disease incidence
State fish kills
Pollution indices
Ichthyoplankton index
Chemical Constituents
Nutrients (N, P, total, soluble)
DO, Alkalinity, conductivity, pH;
nutrient dynamics
Nutrients (N, P, total, soluble)
DO, alkalinity, conductivity, pH;
nutrient dynamics
Habitat and Physical Variables
Temperature
Turbidity (secchi)
Suspended solids
Water depth, area, retention
Substrate characterization
Shoreline development
time
Temperature
Suspended solids
Hydrology
Pool/riffle series
Substrate characterization
Embeddedness
Streambank stability
Width of riparian zone, percent
of stream cover
32
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TABLE 2. GENERAL CHECKLIST OF FISH FIELD EQUIPMENT AND SUPPLIES
and flotation
pack
Boat(s)
Motor(s)
Paddles
Life preservers
cushions
Fire extinguisher,
(US Coast Guard approved)
First aide kit
Running Lights
Air Horn
Camera/film
Maps
Ice chests
Ice
Blue ice, soft
Dry ice
Portable light source
Waterproof notebook
Waterproof pencils, ink pens
Waterproof labels
Arm-length insulated
water proof gloves
Hip boots
Rain gear
Feltsole neoprene chest waders
Paper towels
Aluminum foil
Thermometer
Water chemistry meters or
water test kit
Seechi disk
Glass jars (4L, 2L, 1L)
(chemical samples)
Hand tallys
Tape measure
(100 yd. or meter)
Polaroid glasses
Dip nets
Seines
Gill nets
Trawls
Traps
Hoop nets
Electrofishing gear
Balance (weight scale)
Measuring board (50 cm)
Tubs
Buckets, livewells, coolers
Fish survey data forms
Habitat survey forms
Clip board with cover
Dissecting kit
Plastic bags, various sizes
10% Buffered formalin
(formaldehyde solution)
Ethyl alcohol (ethanol) or
isopropyl alcohol
Distilled or deionized water
Scale envelopes
Divider for measuring
body proportions
Magnifier, pocket
Microscope, field
Dissecting microscope
Microscope slides and cover
Air pump, battery
Calculator
Marker, permanent black
Fish finder
Nylon-mesh fish cage
Sample containers
Data sheets
Patch kit for wader repair
Fiberglass hauling tanks
Anesthesia, MS222 (tricanemethane
sulfonate)
Long forceps
Samll envelopes
Vials or small bottles
Scalpel or knife
Divider, fine-pointed, or
dial caliper
Rule, stainless steel, metric
Other:
33
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record external anomalies found on fish, and the codes are recorded on the
fish field data sheet (Figure 1C),
4.1.5 Habitat Evaluation
4.1.5.1 A general site evaluation of each sampling location should be
conducted during the sample processing because the range of habitats (riffles,
runs, pools) can have a major effect on the data collected. Figure 2 contains
a habitat description sheet for evaluating the surrounding topographical
features and physical characteristics of fish sampling locations. The
information can be used for calculating a Quality Habitat Evaluation Index
(QHEI) described in Ohio EPA (1989) and Rankin (1989). Also, see Hughes et
al. (1986; 1987), Hughes and Larsen (1988), Hunt (1992), Omernik (1987),
Omernick and Gallant (1988), and Section 8, Fish Bioassessment Protocols For
Use In Streams and Rivers.
4.1.6 Regional Reference Site Selection
4.1.6.1 Reference sites should be selected based on the following criteria:
4.1.6.2 Select site using standardized methods.
4.1.6.3 Select site least impacted sites that are typical of the region,
4.1.6.4 Avoid areas below point sources of pollution including known recovery
areas (except large rivers).
4.1.6.5 Avoid areas of obvious habitat modification and nonpoint sources of
pollution or impacts,
4.1,6.6 Select representative sites distributed by stream size.
4.1.6,7 Site can be maintained by continuing to resample the reference site
on a once every ten years basis or less.
4.1.7 Fish Sampling Gear
4.1.7.1 Fish can be collected actively or passively. Active sampling methods
include the use of seines, trawls, electrofishing, chemicals, and hook and
line. Passive methods involve entanglement (gill nets, trammel nets, tow
nets) and entrapment (hoop nets, traps, etc.) devices.
4.1.7.2 The chief limitations in obtaining qualitative and quantitative data
on a fish population are gear selectivity and the mobility and rapid
recruitment of the fish. Gear selectivity refers to the greater success of a
particular type of gear in collecting certain species, or size of fish, or
both. All sampling gear is selective to some extent. Two factors that affect
gear selectivity are: (1) the habitat or portion of habitat (niches) to be
sampled and (2) the actual efficiency of the gear. Another problem is that
the efficiency of gear for a particular species in one area does not
necessarily apply to the same species in another area. The skill and training
34
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A. FISH FIELD DATA SHEET
State or Country:
Locality:
Collection No,
County
Collectors:
Date:
Water :
Shore vegetation:
Aquatic vegetation:
Stream width:
Amax:
Shore:
Bottom:
Weather:
Method of capture:
Water Chemistry:
Depth
Tu
Time:
Temp.:
Air:
Zmean:
pH
Zmean:
Pool current:
Riffle:
Original preservative:
Seechi:
DO Conductivity Salinity
COMMENTS:
Figure 1. Example of a general fish field data sheet.
35
-------
A. FISH FIELD DATA SHEET (CONTINUED)
Page Of Collection No,
State or Country: County
Locality:
Collectors:
Date: Time:
36
-------
B. FISH FIELD DATA SHEET
Coll. No,
State or Country: County
Locality: .
Water:
Vegetation:
Bottom: Temp,: Air;
Shore:
Distance from shore or stream width: Current:
Depth of capture: . Depth of water:
Method of capture:
Collected by: Date:
Orig. preserv.: Time:
Weather:
Figure 1. Example of a general fish field data sheet,
37
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C. FISH FIELD DATA SHEET
Field Crew Collector/Becotdar
River/Stream:
Date:
River Code: ,_ ,
RM:
Distance
~" Sampler Type:
*•• f^.^At..
~~ Depth: _ _n
«tt
~ Da ta Source:
Time of Da v: Paee of
.,_ Location:
Time Fished: '
Total Seconds
Ob<(»rvi»d Flow!
Number of Species:
Anomata:A-awfcorworm; B-Btack spot C-btfas. (ktetormtea: E-voM Int. F-lungus; L-tesons. U-muttipie DELT anomalies. N-tim
P-p*ssites. Y-popey». S^mxastsd, W-swuM scatos; T-umcn. 2-«r*i/ [H-Heavy. L-UgK at* combned witn anomalies A, S, and C]
SPECIES
f WE1OHEO
IOi*L
5pJJffTEO
1
~1
WEIGHT (CRAMS)
Mass Weighing To-
Convention: HJ.U.M i«i
ANOMALIES
536 (u)-
•
-------
SITE DESCRIPTION SHEET
Fish QHEI SCORE: | |
Stream _ , _ __L_: :::: ......... _^_ _ , _ RM _ Date _ River Cod*. __
Location _ _____ _ , _ Ctew:_ _ .
1J SOBSTBATi (Check ONL Y Two Subitrale TYPE BOXES; CfcecJc mil types prttents;
TJPJ POOL RIFFLE POOL RIFFLE SUBSTRATE QUAUp SUBSTRA Tg SCORE- I - 1
QQ-BLD£R/SUBS[10] __ QO-GRAVEL[7] _ _ Subsfratf Origin (Cheek ill) SIK Cover (Cheek Onet ' ........... '
QMOULDERJ8J __ OO-SAND[6j __ Q-LIMESTONE[1p-RlP/RAP[Q] CD-SILT HEAVY [-2JQ-SILT MODERATE [-1]
QQCOSBLEJSl __ QO-BEDROCK(51 __ Q-T1LLS|1] Q-HARDPANJO] O- SILT NORMAL JO] D- SILT FREE [1]
OD-HARDPAN|4] __ O ODETRITUS[3] __ O- SANDSTONE (0) Exleni Of Etnbeddneas tCheek one)
QO-MUOqZ] ___ QD-ARTiRC,[0] __ D-SHALEJ-1] Q— EXTENSIVE [-2] O— MOOERATE[-1]
TOTAL NUMBER OF SUBSTRATE TYPES: Cb 4 [2] D—c- 4 [0] Q-COAL FINES [-2] Q— LOW[0] Q— NONEfl]
NOTE: (Ignore sbdge that ohginatas from point-sources; score is based on natural substrates)
COMMENTS _ . -„.
COVER SCORE: [ \
S} IKST8EAM COVER AMOUJflfChaek ONL Y On* or
TYPE (Chtcfc 4«Trut Apply) check2»nd4KESAG£)
Q-UNDEBCUT BANKS [1] D -DEEP POOLS [2] D -OXBOWS [1] O- EXTENSIVE > 75% [11]
Q-OV£HHAr4G!NG VEGETATION [11 Q-ROOTWADS[1] Q -AQUATIC MACROPHYTES [1] D - MODERATE 25-75% [7]
D -SHALLOWS (IN SLOW WATER) [1] Q-BOULDERS [1] Q -LOGS OR WOODY DEBRIS [1] 0 - SPARSE 5-25% [3]
Q - NEARLY ABSENT < S*{1]
COMMENTS: _ , __ _____
3] CHAfWEL MORPHOLOGY: (Check ONLYQm PER Category OR check 2 ind A YERAGE) CHANNEL: [ _J
SiNUOSfTY Qgya.Q.gMEJg • CHAWiEHZATJQN SJABIirfY,' MQOIF1CAT1ONS/OTHER
Q • HIGH [4J D - EXCELLENT [7] Q - NONE [6] O - HIGH [3J Q - SNAGGING O - IMPOUND,
O- MODERATE [3] D- GOOD [5] Q - RECOVERED [4] Q - MODERATE [2] O- RELOCATION O- ISLANDS
Q-LOW|2] Q-FAIRp] D- RECOVERING [3] Q- LOW [1] Q - CANOPY REMOVAL O - LEVEED
O'-NONE[1) O-?OOR|1J D- RECENT OR NO O-DREDaiNG O • BANK SHAPING
RECOVERY JIJ D - ONE SIDE CHANNEL MODIFICATIONS
COMMENTS: _ _____ ___
4) RIPARIAN ZONE AND BANK EROSION • (check ONE box per bank or check 2 »nd AVERAGE per b»nkl RIPARIAN: j I
*Rw« Rigra Lootung Downstream*
pROglpNf RUNpFF - R_OOD PLAIN OAJAHTY pArjK g
L R (Per Sank) L R (Moit Predominint Per Bank) L R (Per Bank)
aa'-WIDE>SOm[4] DO-FOREST, SWAMP p] Q O-UR8AN OR INDUSTRIAL[0] D Q-NONE OR LITTIE [3]
QD'-MOOERATE 10-50 pj OD-OPEN PASTURE/ ROWCROP[0] 0 O-SHRUB OR OLD RELD[2] Q O-MODERATEI2]
aa--NAHROW5-10m|2] OO-RES!D.,PARK,NEW FIELD [1] DOCONSERV. TILLAGE [ij O O-HEAVYOR SEVEHE[1]
O 3'-VEF!Y NARROW 1-Sm [1] 0 Q-FENCED PASTURE [1] Q O-MININGCONSTRUCTION [0]
COMMENTS: , r—I
POOUSUDE AND RIFFLE/RUN QUALITY POOL: I I
MAX. DEPTH (Check 1i MORPHOLOGY PQQL/RUN/RIFFIE CURRENT VELOCfTY 1—4
O->lm[6) (Check 1} (Check Xtf That Apply)
D-0.7-1 m [4] O'-POOL WIDTH > RIFFLE WIDTH [2) 0'-TORHENTIALJ-1] O"-EDDIES{1]
D-0.4-0.7m PI O-POOL WIDTH. RIFFLE WIDTH [1j O'-FASTJI] O'-tNTERSTITlAH-IJ F5-NO POOL(c|]
Q.<0,4mf1J Cr-POOL WIDTH < RIFFLE W, [0] O'-MODERATE (1J O'-INTERMtTTENTl-t]
Q—<0.2mfPoo».a| Q'-SLOW[1]
COMMENTS:
~~ : RIFFLE: \ |
RIFFLE/RUN pgpTJj BlEFLE/flllM SUBSTRATE RIFFLE/RUN FMBCDDEDNESS I—I
O - GENERALLY >10 em,MAX>50 [4J Q-STABLE (e.g.,Cobbl«, Boulder) |2J D-EXTENSIVE [-1] D-MODERATE[0]
O - GENERALLY >10 em,MAX<50 [3] O-MOD, STABLE (•.8.,P«« Gravel) [1J O-LOW. |1J O-NONEJ2J
O - GENERALLY S-10 cm [t J O-UNSTABLE (Graval.Sand) [0] [5
O - GENERALLY « S cm [Riffle . 0]
COMMENTS _ GRADIENT:
6] Gradient (feet/mile): SPOOL: %HIFFLE: %RUN:
Figure 2. Site description sheet for evaluating the topogeographical features
and physical characteristics of fish sampling location. Adapted
from Ohio EPA (1989).
39
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SITE DESCRIPTION SHEET (CONTINUED)
f Mac.
SEAR
HSTANC
WATSCUWJTY
FRSTPASS
SECOND PASS
TMROPASS
CANOPY (WOPSfl
STIiAH MEASUHEMEHTS: AVERAGE WCTK:.
LENGTVi WtJTH
AVERAGE DEPTH:
,IUX DEPTH
DRAWWQ OF STREAM
Figure 2. Site description sheet for evaluating the topogeographical features
and physical characteristics of fish sampling location. This part
is used to record additional information about the sampling site
and adjacent area. Adapted from Ohio EPA (1989).
40
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TABLE 3, CODES UTILIZED TO RECORD EXTERNAL ANOMALIES ON FISH
Anomaly Code
Description
E
L
T
M
AL
AH
BL
BH
CL
CH
F
I
N
S
P
Y
W
I
Deformities of the head, skeleton, fins, and other
body parts.
Eroded fins.
Lesions, ulcers
Tumors
Multiple DELT anomalies (e.g. lesions and tumors,
etc.) on the same individual fish.
Anchor worm - light infestation: fish with five
or fewer attached worms and/or previous attachment
sites.
Anchor worm - heavy infestation fish with six or
more attached worms and/or previous attachment
sites.
Black spot - Light infestation: spots do not
cover most of the body with the average distance
between spots greater than the diameter of the
eye.
Black spot - Heavy infestation: spots cover most
of the body and fins with the average distance
between spots less than or equal to the diameter
of the eye.
Leeches - Light infestation: fish with five or
fewer attached leeches and/or previous attachment
sites
Leeches - Heavy infestation: fish with six or
more attached leeches and/or previous attachment
sites.
Fungus.
Ich
Blind - one or both eyes; includes missing and
grown over eyes (does not include eyes missing due
to popeye disease).
Emaciated (poor condition, thin, lacking form).
External parasites (other than those already
specified).
Popeye disease.
Swirled scales.
Other, not included above.
'Adapted from Ohio EPA (1989).
41
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of the personnel doing the sampling are also very important in sample
collection.
4,1.7.3 Temporal and spatial changes in relative abundance of a species can
be assessed under a given set of conditions if that species is readily taken
with a particular kind of gear.
4.1.7.4 Passive collection devices usually require little specialized
training to operate and can be used to collect data on relative abundance of
many species. Passive methods, however, are very selective for some species.
Gear type and design used are important in particular habitats to capture
specific species or sizes of fish (Carter, 1954; Hubert, 1983; Starrett and
Barnickol, 1955). Active methods are generally less selective and more
efficient. Although the choice of method depends on the objectives of the
fishery investigation and habitat to be sampled, active methods are generally
preferred. However, the method selected must provide the information required
from the survey or study. The biologist must decide whether he needs
information on standing crop, catch per unit effort, qualitative information
on the fishery, etc., and choose the sampling technique or techniques
accordingly.
4.1.7.5 Sport fish, large specimens, and rare or endangered species should be
identified in the field, measured (standard length, total length, body depth),
examined for external anomalies, and if possible, released unharmed. If the
fish are to be released unharmed, the method and equipment used must be
selected appropriately. Some methods (e.g., gill nets) usually kill the fish.
4.2 Active Sampling Techniques
4.2.1 Seines
4.2.1.1 A haul seine is essentially a strip of strong netting hung between a
stout cork or float line at the top and a strong, heavily-weighted lead line
at the bottom (Figure 3). The wings of the net are often of larger mesh than
the middle portion, and the wings may taper so that they are shallower on the
ends. The center portion of the net may be formed into a bag to aid in
confining the fish. At the ends of the wings, the cork and lead lines are
often fastened to a short stout pole or brail. The hauling lines may be
attached to the top and bottom of the brail by a short bridle. The
quantiative factors of this gear are determined by the total length of the
net, the mesh sizes used in its construction (especially in the bag), and
whether or not the float!ine remains on the surface during operation or under
water with the leadline on the bottom. The size of these seines is usually
determined by how they are retrived and the species sought.
4.2.1.2 Deepwater or haul seining usually requires a boat. One end of one of
the hauling lines is anchored on shore and the boat plays out the line until
it reaches the end. The boat then lays out the net parallel to the beach.
When all of the net is in the water, the boat brings the end of the second
hauling line ashore. The net is then beached as rapidly as possible without
allowing the lead line to come off the bottom.
42
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4.2.1.3 Straight seines (without bags) can usually be handled by two people.
The method of playing out the seine and bringing it in may be similar to the
haul seine or it may be pulled parallel to the shore for some distance before
it is beached. The straight seine is generally used in shallow water where
one member of the party can wade offshore.
4.2.1.4 Bag and straight seines vary considerably in dimensions and mesh
size. The length may vary from 3 to 70 meters, and mesh size and net width
vary with the size of the fish, depth of the water, and the habitat to be
sampled.
4.2.1.5 Nylon seines are recommended because of the ease of maintenance.
Cotton seines should be treated with a fungicide and dried after using to
prevent deterioration. Nylon seines should not be left in the sun for
prolonged periods of time or they will also deteriorate.
4.2.1.6 Seining is not effective in deep water unless the seine is deep
enough to cover the area from surface to bottom. Seining is not effective in
areas that have snags, large rocks and boulders, and sunken debris that may
tear or foul the net. However, in selected areas seines can be very efficient
in sampling fish. Although the results are expressed as number of fish
captured per unit area seined, quantitative seining is very difficult. It
must be applied consistently along several beaches of a waterbody to achieve a
quantitative assessment. The method may be more useful in determining the
variety of fish rather than the number of fish inhabiting the water.
4.2.1.7 Choice of seines will depend on the study design, and sampling
methods and sizes of seines vary with habitat type.
4.2.1.8 Seining should be performed by at least two investigators, but having
more helpers improves sampling effectiveness.
4.2.1.9 In riffles of wadable streams, e.g., the preferred method is the
"foot shuffle" using a 3 m minnow seine with 1/4 inch mesh (6 mm) size. This
kickset method consists of setting the net in the water perpendicular to the
current. Investigators then enter the riffle approximately 3 m upstream from
the net and actively disturb the substrate and overturn rocks or other debris.
The net is then picked up and carefully examined for the presence of fish. In
slower currents, it may be possible to pull the seine downstream, hooking into
the bank after a distance of 5 to 10 m.
4.2.1.10 In pools, because larger seines are preferred, depth of water
usually precludes effective kicksets. In such situations, pools are actively
seined by pulling a 5 m seine with 1/4 inch mesh (6 mm) size through the pool
either perpendicular or obliquely to the bank, or, in the case of very quiet
water, upstream or downstream and parallel to the bank prior to hooking into
shore and examining for fish.
4.2.1.11 Continue seining until two riffles and two pools or, in the absence
of discrete habitats, a segment of at least 200 m has been sampled. Distance
sampled should not exceed 500 m. Record total time spent collecting.
43
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4.2.1.12 Record all information on field data sheets. Specimens kept for
later identification or for voucher specimens should be preserved in 10%
formalin solution (see Section 5, Fish Specimen Processing) and kept in
separate jars by habitat type with inner and outer waterproof labels. Labels
should contain locality data, habitat type, date, collectors names, and study
collection numbers from the field sheets for that site.
4.2.2 Trawls
4.2.2.1 Trawls are specialized submarine seines used in large, open water
areas of reservoirs, lakes, large rivers, estuaries, and oceans. They may be
of considerable size and are towed by boats at speeds sufficient to overtake
and enclose the fish. Four basic types are available: (1) the beam trawl
used to capture bottom fish, (2) the otter trawl used to capture near-bottom
and bottom fish, (3) the mid-water trawl used to collect schooling fish at
various depths, and (4) surface tow nets used to collect fish at or near the
surface. These trawls can be very effective on selected bottom, mid-water and
surface oriented species at specific life history stages.
4.2.2.2 The beam trawls (Figure 4) have a rigid opening and are difficult to
operate from a small boat. Otter trawls (Figure 5) have vanes or "otter
boards", which are attached to the forward end of each wing and are used to
keep the mouth of the net open while it is being towed. The otter boards are
approximately rectangular and usually made of wood, with steel strapping. The
lower edge is shod with a steel runner to protect the wood when the otter
board slides along the bottom. The leading edge of the otter trawl is rounded
near the bottom to aid in riding over obstructions. The towing bridle or warp
is attached to the board by four heavy chains or short heavy metal rods. The
two forward rods are shorter so that, when towed, the board sheers to the
outside and down. Thus, the two otters boards sheer in opposite directions
and keep the mouth of the trawl open and on the bottom. Floats or corks along
the head rope keep the net from sagging, and weights on the lead-line keep the
net on the bottom. The entrapped fish are funneled back into the bag of the
trawl (codend). The size of the mesh in the codend (bag end of a net) will
determine the species and life history stages caught.
4.2.2.3 The midwater trawl resembles an otter trawl with modified boards and
vanes for controlling the trawling depth. Such trawls are cumbersome for
freshwater and inshore areas, but can be used very effectively in marine and
estuarine waters. Surface townets have been used very effectively for
emigrant juvenile salmonids in northwest and Alaska estuaries for monitoring
year class abundance.
4.2.2.4 A popular, small trawl consists of a 16 to 20 foot (5 to 6 meters)
headrope, semiballoon modified shrimp (otter) trawl with 3/4 inch (1.9 cm) bar
mesh in the wings and cod end. A 1/4 (0.6 cm) bar mesh liner may be installed
in the cod end if smaller fish are desired. This small trawl uses otter
boards, the dimensions of which, in inches, are approximately 24 to 30 (61 to
76 cm) x 12 to 18 (30 to 46 cm) x 3/4 to 1/4 inches (0.9 to 3.2 cm), and the
trawl can be operated out of a medium-sized boat.
44
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Pole or Brail
Figure 3. The Common Haul Seine. Modified from Dumont and Sundstrom (1961)
Figure 4. The Beam Trawl. Modified from Dumont and Sundstrom (1961).
45
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Figure 5. The Otter Trawl. Modified from Dumont and Sundstrom (1961).
4.2.2.5 Trawling data are usually expressed in weight numbers, species,
etc.of catch per unit of time.
4.2 2.6 The use of trawls requires experienced personnel. Boats deploying
large trawls must be equipped with power winches and large motors. Also,
trawls can not be used effectively if the bottom is irregular or harbors snags
or other debris. Trawls are used to gain information on a particular species
of fish and an overally estimation of fish populations and communities. See
Hayes (1983), Massman et al. (1952), Rounsefell and Everhart (1953), and Trent
(1967) for further information on trawls.
4.2.2.7 In selected studies a plankton net may be used as a trawl. Larval
and young fishes can be collected at the surface and bottom with a 1 meter
plankton net by trawling a transect with a predetermined time frame (say ten
minutes). A plankton sled can be used to hold a meter plankton net towed at
the bottom while a sidearm can be used at the surface (Dovel, 1964). A
digital flowmeter can be mounted in the mouth of the net to determine the
amount of water strained. Large numbers of plankton can be collected in a
short time by using a Miller high-speed sampler. Another sampler type, the
bongo net, is a pair of nets held side-by-side in a frame and is towed by a
cable that attaches to the frame between the two nets. Bongo nets are good
because they can be used off ships at high speed, can be used to sample the
horizontal layer of the water column, and can be used to get replicate samples
at the same time.
46
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4.2.2.8 The larval stage by some individuals is considered the period from
time of hatching until the attainment of the adult fin-ray complement,
ossification of spines or rays and the inception of scale development.
Mansuetti and Hardy (1967) defined the "larval" stage as the period from the
disappearance of the yolk sac until the development of the adult fin-ray
complement.
4.2.3 Several companies sell a variety of fish nets, seines, traps, trawls,
etc.:
1. Sterling Marine Products, 18 Label Street, Montclair, NJ 07042,
Telelphone (201) 783-9800 or Jonesport, Maine 04649, Telephone (207)
497-5635
2. Nylon Net Company, 615 E. Bodley Avenue, P.O. Box 592, Memphis, TN
38101, Telephone (901) 774-1500, FAX (901) 774-8130.
3. Memphis Net and Twine Company, Inc., 2481 Matthews Avenue, P.O. Box
8331, Memphis, TN 38108, Telephone (901) 458-2656, FAX (901) 458-
1601.
4. Nichols Net and Twine Company, Inc., R.R.3 Bend Road, East St.
Louis, IL 62201, Telephone (618) 876-7700.
4.2.4 Horizontal Ichthyoplankton Tow-Net
4.2.4.1 The larval fish sampler (Figure 6) consists of a modified bridle,
frame, and net system with an obstruction-free opening. The tow net is easy
to handle, and it is small enough for use on boats 4 m or larger in length.
The tow net features a square net frame attached to a 0.5 m diameter cylinder-
on-cone plankton net with a bridle. This design eliminates all towing
obstructions forward of the net opening; in addition, it significantly reduces
currents and vibrations in the water directly preceding the net. See
Subsection 4.2.4.2 for the design and construction details of the horizontal
ichthyoplankton tow-net. With the aid of a stanchion and winch assembly, one
person can easily sample any stratum from near surface to near bottom in lakes
and rivers. The cylinder-on-cone net is self-washing while it is being
fished, and only the last 20 cm needs to be rinsed to concentrate the sample
in the collecting bucket. The system is self closing during deployment and
retrieval. During deployment, the towing cable is payed out at approximately
the same speed that the vessel is moving forward. This allows the weighted
net to rapidly descend, with the net mouth in the vertical plan, while
collapsing the net body and thus preventing the net from fishing. When the
net has reached the desired fishing depth, the release of the towing cable is
stopped and the net begins fishing (Figure 6). Prior to retrieval, the vessel
is stopped, and the vertical orientation of the net mouth and rapid lifting
causes the net body to collapse, preventing the net from fishing. Nester
(1987) and Nester (1992, personal communication) reported that the tow net
system is effective in collecting all lentic fish larval species at sampling
depths ranging from surface to 10 m and can easily be used at greater depth.
4.2,4.2 The 6.3-mm galvanized steel towing cable (1) is connected to the
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center of the fore-bridle (2) with a 76.2-mm heavy duty snap swivel. A 25.4-
mm thimble is permanently fixed with 3.2-mm cable clamps in the center of the
3.2-mm galvanized steel cable fore-bridle. The spreader bar (3) is
constructed of 9.6-mm cold-rolled steel with two 38.8-mm clevises welded in
place at either end. Side cable (4 and 5) of 3.2-mm galvanized steel are
connected to the spreader bar clevises and to clevises welded to each corner
of the net frame (6). The net frame is constructed of 9.6-mm cold-rolled
steel heated, bent to form a 53-cm square, and closed by welding. Corner
supports (7) provide additional strength and attachment points for netting.
Two flowmeter support brackets (8a and 8b), to which flowmeters (9) are
attached, are welded to the net frame and corner supports. Each bracket is
bent at two 45° angles, so that the free end is about 5-cm behind the plane of
the mouth of the net. Stainless steel support cable (10) is passed through a
pair of 116-mm holes drilled 3-cm apart in the free end of the bracket to
support the flowmeter. Nylon cord is used to lash the 0.5-m-diameter brass
net ring (11) to the net frame and corner supports. The net bucket is secured
to the cod end (bag end of a net) of the net with a hose clamp. Cables (12)
supporting the 1-kg depressor plate (13) are attached to the lower corners of
net frame with 3.2-mm cable clamps.
V
12
13
Figure 6. Horizontal Ichthyoplankton Tow-Net. Attitude of the modified
bridle, frame, and net of the sampling system and diagram of the
construction details. Numbers are referred to in Subsection
4.2.4.2. From Nester (1987).
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4.2.4.3 The 0.5-m plankton net, a cylinder-on-cone configuration, is
constructed of 0.335-mm mesh to the dimension given by O'Gorman (1984). A net
of this type should have superior sustained filtration ability because of the
high ratio of open mesh area to mouth area (6.3:1) and because oscillations
of the cylindrical portion seemingly help clean the net during a tow (Tranter
and Smith, 1968).
4.2.4.4 Commercial ichthyoplankton nets are available with threaded cod ends.
Plastic cod end jars can be easily screwed into these and when the sample is
finished being collected and preserved, a lid is screwed onto the jar and a
new jar added to the net. These allow for rapid sample handling and decreased
time. These have been found to be important if there are a lot of samples to
be taken and numerous collecting sites.
4.2.4.5 Information on collecting and processing fish eggs and larvae
are found in Simon (1989), Snyder (1983), and marine recommendations are
provided by Smith and Richardson (1977).
4.3 Electrofishing
4.3.1 Electrofishing is an efficient capture method that can be used to
obtain reliable information on fish abundance, length-weight relationships,
and age and growth of fish in most streams of order 6 or less (Platts et al.,
1983 and Plafkin et al., 1989). Note: Individuals involved in electrofishing
must have completed a certified training course in electrofishing or have been
trained by someone certified and experienced in electrofishing. This
subsection provides some general principles and guidelines for understanding
electrofishing. Electrofishing is a method for collecting fish using
electricity. Either alternating (AC) or direct (DC) electrical current can be
used. Most electrofishing in freshwater is done with pulsed DC electrical
current equipment. In a boat-rigged shocker (boom shocker) or airboat, one or
two people net the fish and another operates the boat and equipment. The fish
are nearly always driven into cover as a result of electric stimulus making
them difficult to capture. Once driven from cover, the fish are kept within
effective range of the electrical field and are immobilized making it possible
to pick them up with long-handled dip nets. Electrical dislodgement and
immobilization of fish together result in more consistent success under
varying conditions than ordinary seining. However, if target assemblage is
common species, seining may be just as effective. For a discussion of the
general principles and guidelines for electrical fishing, see below and Cowx
(1990), Cowx and Lamarque (1990), Cross and Stott (1975), Dauble and Gray
(1980), El son (1950), Friedman (1974), Hartley (1980), Kolz (1989), Kolz and
Reynolds (1989a, 1989b), Loeb (1955), Novotny and Priegel (1971, 1974), Ohio
EPA (1987a, 1987b, 1989), Reynolds (1983), Sharpe (1964), U.S. Fish and
Wildlife Service (1991), Vincent (1971) and Section 12, Fisheries
Bibliography, 12.2 Electrofishing.
4.3.2 The decision to use electrofishing equipment (or electrofishers) will
depend on size of site, flow, turbidity, and conductivity. If conductivity is
below lOO^S (micro seimens) or if water is too turbid to locate stunned fish,
the investigators should consider other sampling devices (e.g., seines).
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4.3.3 A choice of electrofishing equipment will depend on size of stream and
access to stream fronf road. If a site is wadeable and close to the road, use
Sportyak-mounted, generator unit or equivalent. If access is problematical,
use a back pack unit. For safety reasons, it is Important too always wear
waders and lineman's Insulated (or Playtex Living) gloves when working with
electricity in water. At least two individuals for safety reasons (see Section
3, Safety and Health) are need when electrofishing. Always wear polarized
sunglasses to aid vision.
4.3.4 Electrofishing efficiency can be placed in one of three categories:
fish characteristics, habitat characteristics, and operating conditions. For
a discussion of these three categories, see Reynolds (1983).
4.3.5 It is also recommended that anyone involved in electrofishing must take
a U.S. Fish and Wildlife training course in electrofishing, or they must be
trained by someone experienced in electrofishing.
4.3.6 Electrofishing Equipment (Electrofishers)
4.3.6.1 Electrofishing today is done by wading in shallow streams and using
electric seines, backpacks, tow barges, longlines, etc. or in deep streams and
rivers with electrofishing boats.
4.3.6.2 Typically a flat-bottom boat (usually 12 to 18 ft) is used for
electrofishing in waters too deep for wading (Novotny and Priegel, 1974).
Paired booms, (length vary according to boat size), protrude in front of the
boat and are adjustable for height and spacing by means of lock-in
adjustments. The electrode system should also be adjustable, i.e., operating
with one or both anodes, varying the number of dropper electrodes, varying the
exposure on the dropper electrodes, and alternating the polarity (Figure 7).
Figure 7. Typical Boom Shocker. Photo courtesy of Wisconsin Department
of Natural Resources.
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4.3.6.3 The electrofishing power unit may consist of a 240-volt or 500-volt,
2000 watt, heavy-duty generator and an electrical control section consisting
of a modified, commercially-available, variable-voltage pulsator. The
frequency of the cycles/second or Hz is not a critical factor. For AC
electrofishing, 60, 180, and 400 Hz have been used with similar success. An
electric control section permits the selection of AC voltage from 50-700 and
DC voltage from 25 to 350; furthermore, it permits control of the electrical-
field size which is dictated by the variable conductance of dissolved minerals
in the water. The power equipment is similar in both boat shocking and stream
shocking, but is portable in the latter. The literature indicates that DC
electrofishing is the most comprehensive and effective, single method for
collecting fish in rivers and streams (Gammon, 1973, 1976; Novotny and
Priegel, 1974, and Ohio EPA, 1987b, 1989; Vincent, 1971).
4.3.6.4 Backpack electrofishers are entirely housed in a weatherproof
metallic container that is fastened securely to a comfortable pack frame
(Blair, 1958; Braem and Ebel, 1961; McCrimmon and Berst, 1963; Reynolds, 1983;
Sharpe and Burkhard, 1969). Backpack shocker units can be purchased
commercially. The power source is either a 12-volt (deep charge battery or a
small 115-volt AC generator). The electrode system is hand-held and must be
insulated from the operator by handles 1.5-3.0 m long, preferably made of
fiberglass. A horizontal ring or spatula electrode attached to the end of the
handle is easiest and most effective to use. Positively activated switches on
each electrode handle are an important safety features. Both backpack, tote
barge, and boat mounted shockers are available from U.S. manufacturers in a
variety of models (see Subsection 4.3.13).
4.3.6.5 Other electrofishing devices include: tote barges/sport yaks (Ohio
EPA, 1987b, 1989); longlines (Ohio EPA, 1987b); electric trawls (AC) (Haskell
et al., 1955); and (Loeb, 1955); electric seines (Funk, 1947; Holton, 1954;
Larimore, 1961; Bayley et al., 1989); and a fly-rod electrofishing device
employing alternating polarity current (Lennon, 1961). After reviewing the
literature, the user must decide which design is most suitable to the
particular needs of the study.
4.3.6.6 Decision on the use of AC, DC, pulsed DC or alternate polarity forms
of electricity and selection of the electrode shape, electrode spacing,
voltage and proper equipment depends on the resistance, temperature and total
dissolved solids in the water. Light-weight conductivity meters are
recommended for field use. Lennon (1959) provides a comprehensive table and
describes the system or combination of systems that worked best for him.
Novotny and Priegel (1974) provide improved designs to increase the
effectiveness of boom shockers.
4.3.6.7 Rollefson (1958, 1961) tested and evaluated AC, DC, and pulsated DC,
and discussed basic electrofishing principles, wave forms, voltage-current
relationships, electrode types and designs and differences between AC and DC
and their effects in hard and soft waters. He concluded that pulsated DC was
best for power economy and fishing ability when used correctly. Haskell and
Adelman (1955) found that slowly pulsating DC worked best in leading fish to
the anode. Pratt (1951) also found the DC shocker to be more effective than
51
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the AC shocker. Frankenberger (1960) and Latta and Meyers (1961) used a DC
shocker, and tarimore et al. (1950) used AC current in boat shocking. Stubbs
(1966) used DC or pulsed DC and had his aluminum boat wired as the negative
pole.
4.3.6.8 Fisher (1950) found that brackish water requires much more power
(watts) than freshwater even though the voltage drops may be identical.
Lennon and Parker (1958) and Seehorn (1968) recommended the use of an
electrolyte (salt blocks) when sampling in some soft waters to produce a large
enough field with the electric shocker.
4.3.6.9 Novotny and Priegel (1974) provided operational guidelines to
increase the effectiveness of boom shockers. They suggest that in the
operation of DC or pulsed DC it is important that the electrofishing boat move
much more slowly than in using AC. In general, AC operation is preferable at
night in shallow clear water where visibility is no problem, and it is not
necessary to attract fish from cover. Pulsed DC is effective in deep or
turbid water where fish must be drawn from cover and collected by long-handled
dip nets.
4.3.7 Areas considered as problems in boat electrofishing are (Novotny and
Priegel, 1974):
4.3.7.1 Range limitations (distance at which fish are affected).
4.3.7.2 Water conductivity (difficulty in attaining sufficient current in
water of low conductivity.
4.3.7.3 Bottom materials (reduces effectiveness of electrofishing by highly
conductive bottom material).
4.3.7.4 Water depth (difficulty capturing immobilized fish at depths beyond
0.9 - 1.2 m) due to visibility, length of dipnet handle, etc.
4.3.7.5 Water clarity and vegetation (these factors restrict visibility).
4.3.7.6 Water temperature (best response depends on the species and water
temperature).
4.3.7.7 Fish mortality (much higher with AC electricity than DC or pulsed
DC).
4.3.7.8 Fish size (selectivity of size) is not much of a problem with modern
electrofishing units.
4.3.7.9 Fish species (selectivity for species-swimming ability).
4.3.7.10 Equipment and operating problems (inadequate lighting, power,
voltage controls, instrumentation, electrode design, etc.).
4.3.7.11 Day and night sampling (some species sampled better during the day
than night and vice versa) (Sanders, 1991; 1992).
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4.3.7.12 Novotny and Priegel (1974) were able to overcome many of the above
problems. These same problems are also encountered in stream electrofishing
(Novotny and Priegel, 1971).
4.3.8 Safety
4.3.8.1 In order for electricity to flow, electricity needs a complete
electrical circuit, moving from the anode to the cathode. Therefore, the only
way an individual can get shocked is if they become part of the circuit.
During electrofishing, the water becomes the connection that completes the
circuit between the anode and the cathode. You must, therefore, be
electrically insulated from the water and the electrodes of the electroshocker
(electrofisher). Otherwise, you become part of the circuit and will get a
shock.
4.3.8.2 Novotny and Priegel (1974) and Ohio EPA (1989, 1990) give a complete
description of an electrical safety disconnect system and discuss electrical
safety and safety regulations. An 18 foot boat provides a greater margin of
safety in rough water, and a safety railing surrounding the front deck and
extending along each side of the boat affords protection of the operators
against the hazard of falling over-board into the electric field near the
boat.
4.3.8.3 Floor mat switches or foot pedals with non-skid surfaces should be
permanently installed on the front deck. Thus, each operator must be in
position before the system is energized. Likewise, a throw switch should be
installed on the rear seat for the outboard motor operator.
4.3.8.4 When metal booms are used an electrical ground wire terminated with a
battery clamp should be provided to assure a positive electrical ground for
each boom.
4.3.8.5 All electrical circuits should be enclosed in metal conduit with a
separate conduit system for the main power (high voltage) circuits, auxiliary
power and safety circuits (low voltage). Watertight junction boxes should be
used throughout the electrical system.
4.3.8.6 Because the nets used to capture fish must be dipped into the water
near the electrodes, it is very important that the net handles be constructed
of materials with good electrical insulating properties. Epoxiglass
insulating materials used on electricians tools are the best material.
Fiberglass covered metal can cause accidents if the fiberglass covering is
damaged, allowing contact between the operator and the metal handle. The
operator must wear rubber gloves.
4.3.8.7 All leads associated with the generator are carefully insulated.
Generally, AC or DC, used in electrofishing provides more than enough voltage
and current to shock and electrocute a person.
4.3.9 In a boat shocking operation the following safety precautions should be
observed:
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4.3.9.1 Wear U.S. Coast Guard approved life jackets.
4.3.9.2 Wear felt sole neoprene waders or hip boots and insulated arm length
gloves.
4.3.9.3 Avoid excess fatigue and be constantly alert.
4.3.9.4 Authorize one person to be in charge.
4.3.9.5 Instruct all personnel in the fundamentals of electricity.
4.3.9.6 Thoroughly familiarize all persons with all phases of the equipment
and its operation.
4.3.9.7 Make sure that all equipment is in good condition and properly used.
4.3.9.8 Make sure that there is a first aid kit and fire extinguisher on the
boat.
4.3.9.9 Know how to administer first aid treatment for electrical shock.
4.3.9.10 Never operate electrofishing equipment if you have any prior heart
ailment.
4.3.10 The following things must be done to prevent electrical shock when
using electrofishing equipment:
4.3.10.1 Use water-tight, preferably chest or waist-high, waders (neoprene
waders with felt-sole boots). If the waders or wading boots become wet
inside, stop electrofishing and let them dry out thoroughly before
electrofishing again. Wet wading boots can conduct electricity.
4.3.10.2 Use water-tight lineman's insulated gloves that cover up to at least
the elbows. If they get wet inside, stop electrofishing and let them dry out
completely before continuing electrofishing.
4.3.10.3 The individual doing the electrofishing must take care not to let
the anode come into contact with anyone while the unit is active. In
addition, one must make sure and be aware that anyone in or near the water is
electrically insulated with wading boots and gloves.
4.3.11 Electrofishing procedures for use in wadable streams
4.3.11.1 The sampling gear should consist of backpack electrofishing
equipment supplemented by block netting and seining in habitats where flow,
substrate, and structure affect capture of benthic fish species.
4.3.11.2 The investigator(s) should follow project plans, standard operating
procedures (SOPs), and safety for electrfishing in wadable streams and rivers.
4.3.11.3 Decision to use electrofishing equipment will depend on size of
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site, flow, and turbidity. If flow is too high, site to deep, or water too
turbid to locate stunned fish, the investigators may consider use of seine
only. This is a safety decision.
4.3.11.4 Once the sampling site has been located, determine the fish sampling
reach as a function of mean channel width taken at the site (20-30 channel
widths). The sampling site may serve as the midpoint of the sample reach.
The investigator should walk the length of the sample site to determine pool
depths, habitat composition, barriers, and obstructions which may impede or
aid in fish capture. Also, determine if reach requires block nets be placed
at upstream and downstream ends of the stream (e.g., where sample reach is a
large continuous pool).
4.3.11.5 Set the electrofishing unit to 300 VA and pulsed DC. Based on
stream conductivity, select initial voltage setting. Determine that all
crewmembers are wearing waders, gloves, are clear of the anode. Start
generator, set timer, and depress switch to begin fishing. Starting at the
bottom of the most downstream riffle, pool, or other habitat type in the
sampling reach, fish in an upstream direction, parallel to the current.
Adjust voltage and waveform output according to sampling effectiveness and
incidental mortality to specimens. Voltage gradients of 0.1 to 1.0 volts/cm
are effective for stunning fish. These gradients can be maintained in
freshwater of normal conductivity (100-500 micromhos/cm) by adjusting circuit
voltage to produce a current of 3-6 amperes (Reynolds, 1983).
4.3.11.6 With switch depressed, sweep electrodes from side to side in the
water in riffles and pools. Sample available cut-bank and snag habitat as
well as riffles and pools.
4.3.11.7 Netters follow along behind person operating shocker and net stunned
fish which are then deposited in separate buckets or holding tanks based on
habitat from which fish are collected. Minnow seines (4 m x 2 m x 0.5 cm) and
kick nets (2 m x 2 m x 0.5 cm) may be used to block in riffles, polls, and
snags.
4.3.11.8 Depending on the study design, fish may be collected according to
time and distance criteria. The collection time should be no less than 45
minutes and no greater than 3 hours for a distance of between 150 - 500 m in
order to obtain replicate samples from two riffles and two pools, or in the
absence of discrete habitat types, a segment of at least 200 m of stream has
been sampled. Homogeneous (or large systems) without clearly defined habitat
types should be sampled wherever best fish habitat is found. Distance sampled
should not exceed 500 m. Record total time spent collecting.
4.3.11.9 Record all information on field data sheets. Sport fish, large
specimens and threatened and endangered species should be identified in the
field, measured (standard length, total length, body depth), examined for
external anomalies, and released unharmed. All other specimens should be
preserved in 10% formalin solution (see Section 5, Specimen Processing
Techniques) and kept in separate jars by habitat type with inner and outer
waterproof labels. Labels should contain locality data, habitat type, date,
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collectors names, and study collection numbers from the field sheets for that
site.
4.3.12 Standard operating and safety procedures for commercial shocker boat
should be followed in beatable streams and rivers.
4.3.13 Companies that sell a variety of electroshocking equipment
(electrofishing boats, boat outfitting electrofishing kits, electrofishing
tote barges, backpack electrofishing units, electrofishers, etc.):
1. Coffelt Manufacturing, Inc., P.O. Box 1059, Flagstaff, AZ 86002 or
1311 E. Butler Avenue, Building B, Flagstaff, AZ 86011, Telephone
(602) 774-8829
2. Smith-Root, Inc., 14014 N.E. Salmon Creek Avenue, Vancouver, WA
98686, Telephone (206) 573-0202
4.4 Chemical Fishing (Ichthyocides)
4.4.1 Fish toxicants for sampling fish populations are a common practice in
impounded waters and streams throughout the United States. Only registered
fish chemical toxicants should be used in collection fish populations. The
Federal and State rules should be check prior to use because they continually
are updated and subjected to change. The decision to use a chemical toxicant
should be based not only on the efficacy of the toxicant, but also on its
persistence in the environment, toxicity to other animals, and whether it is
deleterious to man. Fish toxicants for reclamation are thoroughly reviewed by
Lennon et al. (1971), and papers addressing their use in sampling are found
throughout the literature. Additional information on sampling fish
populations with toxicants is found in APHA (1992), ASTM (1992), Bone (1970),
Boccardy and Cooper (1963), Davies and Shelton (1983), Hocutt et al. (1973),
Hooper (1960), Marking (1992), Meyer et al. (1976), Platts et al. (1983),
Schnick (1974), Schnick and Meyer, (1978), and Section 12, Fisheries
Bibliography, Subsection 12.3, Chemical Fishing.
4.4.2 Chemicals used in fish sampling include rotenone, cresol, copper
sulfate, antimycin A, and sodium cyanide. The ideal ichythocide indicated by
Hendricks et al. (1980) is (1) nonselective; (2) easily, rapidly, and safely
used; (3) readily detoxified; and (4) not detected and avoided by fish.
4.4.3 When using an ichthyocide, care must be taken to ensure that it will be
used correctly and approval for use must be obtained from proper Federal and
State authorities. Hendricks et al. (1980) reported that improper application
of rotenone can have disastrous effects downstream.
4.4.4 Rotenone (Derris or Cube roots) has generally been the most acceptable
because of its high degradability, freedom from such problems as precipitation
(as with copper sulfate), and relative safety for the user.
4.4.5 Pesticides, copper sulfate, cresol, and other chemicals have been used
as fish toxicants, but they are toxic to humans, may add taste or odor to the
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water, have a slow rate of detoxification, may be toxic to other organisms,
and, therefore, should not be used for sampling purposes.
4.4.6 Antimycin A has been registered by the Governments of the United States
and Canada as a fish toxicant since 1966. The dry formulation is known as
"Fintrol" and has been registered by a commercial company. Field trials have
been made and reported by the U.S. Fish and Wildlife Service. Successful
usage has been reported over a wide range of water qualities and water
temperatures. It is effective on fish at concentrations of 1 part per billion
and less but is reported to be relatively harmless to plants, insects,
mammals, and birds.
4.4.7 Rotenone is also registered for fishery use by the U.S. Environmental
Protection Agency according to the Federal Environmenal Pesticide control Act
(Schnick and Meyer, 1978). Rotenone, obtained from the derris root (Deguelia
eTliptica, East Indies) and cube root (Lonchocarpusw m'cour, South America) in
the family Leguminosae, has been used intensively in fisheries work throughout
the United States and Canada since 1934 (Krumholz, 1948). Rotencne kills fish
by blocking oxygen uptake, and the fish suffocate. The toxicity of Rotenone
is a function of the species, size of fish, and water temperature. The pH,
dissolved oxygen, and suspended particulate matter in the water can also
affect its toxicity. It is effective in a short time period. Also, it has
low toxicity to birds and mammals (Hendricks et al., 1980). Davies and
Shelton (1983) reports that Rotenone at concentrations of 1.0 to 2.0 mg/L is
lethal to zooplankton and many aquatic invertebrates, but the effects is short
term. Although toxic to man and warm-blooded animals (132 mg/kg), rotenone
has not been considered hazardous in the concentrations used for fish
eradication (0.025 to 0,050 ppm active ingredient) (Hooper, 1960), and has
been employed in waters used for bathing and in some instances in drinking
water supplies (Cohen et al., 1960, 1961). Adding activated carbon in the
water treatment process not only effectively removes rotenone, but also
removes the solvents, odors, and emulsifiers present in all commercial
rotenone formulations.
4.4.8 Rotenone obtained as an emulsion containing approximately 5% active
ingredient, is recommended because of the ease of handling. It is a
relatively fast acting toxicant. In most cases, the fish will die within 1 to
2 hours after exposure. Rotenone decomposes rapidly in most lakes and ponds
and is quickly dispersed in streams. In warm water lakes or streams at summer
water temperatures, toxicity lasts 24 hours or less. In cold water lakes
toxicity may last for 5 to 30 days. Detoxification is brought about by five
principal factors: dissolved oxygen, light, alkalinity, heat, and turbidity.
Of these, light and oxygen are the most important factors.
4.4.9 Although the toxicity threshold for rotenone differs slightly among
fish species, it has not been widely used as a selective toxicant. It has,
however, been used at a concentration of 0.1 ppm of the 5% rotenone emulsion
to control gizzard shad (Bowers, 1955). For most species the toxicity of
rotenone is greatest between 10°C (50°F) and 23.9°C (75°F), and a 0.5 mg/L of
formulation (0.025 mg/L of rotenone) kills most fish species. The toxicity
drops as temperature decreases. Formulation of 1.0 to 2.0 mg/L is usually
used to insure a complete kill, and blocking nets should be used in the
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sampling area to ensure the desired catch. Sensitivity to rotenone varies
considerable among species and among life stages within a species (Holden,
1980). The toxicity is affected by temperature, pH, oxygen concentration, and
light (Hendricks et al., 1980; Holden, 1980). USEPA (1978) recommends a
concentration of 0.1 mg/L for sensitive species, and a concentration of 0.7
mg/L is recommended if bullheads and carp are present.
4.4.10 Chemical sampling is usually employed on a spot basis, e.g., a short
reach of river or an imbayment of a lake or reservoir. A concentration of 0.5
ppm active ingredient (1/2 gal. 5% rotenone/acre ft.) will provide good
recovery of most species of fish in acidic or slightly alkaline water (Table
4). If bullhead and carp are suspected of being present, a concentration of
0.7 ppm active ingredient is recommended. If the water is turbid and strongly
alkaline and resistant species (i.e., carp and bullheads) are present, use 1-2
ppm. However, caution is advised because rotenone dispersed into peripheral
water areas may kill fish as long as the concentration is above 0.1 ppm. When
rotenone is used in an embayment, some sort of blocking system should be in
place to prevent fish in the area from escaping. Block seines or divers have
been successfully used in past studies. Chemical blocks can be used but are
recommended only when nets or divers cannot be successfully employed.
4.4.11 A very efficient method of applying emulsion products to lake waters
and embayments is to pump the emulsion from a drum mounted in the bottom of a
boat. The drum should be equipped with an outside tube, mounted on the drum
and calibrated to indicate how fast the chemical is being pumped out of the
barrel. The emulsion is suctioned out by a venturi pump (Amundson Boat
Bailer) clamped on the outboard motor. The flow can be metered by a valve at
the drum hose connection. This method gives good dispersion of the chemical
and greater boat handling safety since the heavy drum can be mounted in the
bottom of the boat rather than above the gunwales as required for gravity
flow.
4.4.12 If spraying equipment is used, it will vary according to the size of
the job. For small areas of not more than a few acres a portable hand pump
ordinarily used for garden spraying or fire fighting is sufficient. Some
individuals have successfully used a back-pack fire pump to collect fish
samples from small streams or sections of streams, A mixture of one quart
rotenone in five gallons of water is applied in small amounts.
4.4.13 A power-driven pump is recommended for a large-scale or long-term
sampling program. The capacity of the pump need not be greater than 200 L per
minute. Generally, a 1-1/2 h.p. engine is adequate. The power application of
rotenone emulsives requires a pressure nozzle, or a spray boom, or both, and
sufficient plumbing and hose to connect with the pump. The suction line of
the pump should be split by a "y" to attach two intake lines. One line is
used to supply the toxicant from the drum, and the other line to supply water
from the lake or embayment. The valves are adjusted so that the water and
toxicant are drawn into the pumping system in the desired proportion and
mixed. A detailed description of spraying equipment can be found in
Mackenthun (1969); Mackenthun and Ingram (1967).
4.4.14 A drip method is generally used to dispense rotenone to a flowing
58
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system. Select a 30 to 100 meter reach depending on the depth and width of
the stream; measure the depth of the section selected, calculate the area and
flow and determine the amount of chemical required (Table 5). Block off the
area upstream and downstream with seines. Position containers of liquid
rotenone at the upstream end of the stream reach to be sampled. Nozzles on
the containers must be metered to deliver the predetermined amount of rotenone
to the stream. For additional details concerning the use of a delivery system
for the drip method and nomographs for calculating the amount of toxicant
refer to Price and Haus (1963) and Davies and She!ton (1983). The toxic
effect of rotenone can be eliminated almost immediately with potassium
permangate (KMnOJ at 1 mg/L for each 0.05 mg/L of rotenone (Lawrence, 1955,
1956; Davies and Shelton, 1983). In lentic waters, the potassium permangate
needed to oxidize rotenone is equal to the amount of rotenone applied plus the
chlorine demand of the water. In lotic waters the amount has been estimated
as 2.5 mg/L per cubic foot per second during the entire time the rotenone is
passing through the neutralization point (Platts et al., 1983). Also,
potassium permangate is considered toxic to some fish species at 3 ppm.
Potassium permanganate is also hazardous to apply, and nose, throat, and eye
protection should be exercised by anyone working with it.
4.4.15 The following company sells aquaculture, quality manufactured drugs,
chemicals, biological, scientific supplies, and fish farming equipment:
Argent Laboratories
9702 152nd Avenue Northeast
Richmond, WA 98052, Telephone (206) 885-3377
4.5 Hook and Line
4,5.1 Fish collection by hook and line can be as simple as using a hand-held
rod or trolling baited hooks or other lures, or it may take the form of long
trot lines or set lines with many baited hooks. In generally, the hook and
line method is not acceptable for conducting a fishery survey, because it is
too highly selective in the size and species captured and the catch per unit
of effort may be low. Although it can only be used as a supporting
technique, it may be the best method to obtain a few adult specimens for
contaminant analysis, etc., when sampling with other gear is impossible.
4.5.2 A variation of this is "jug fishing" where a short drop line of 2-3
feet with a baited hook is attached to a jug or can and allowed to drift
downstream. This is a particularly effective way of sampling catfish.
4.6 Passive Sampling Techniques
4.6.1 Passive sampling devices and techniques (Hubert, 1983) can be used to
supplement boat electrofishing data in lakes, reserviors, large rivers,
estuaries, marshes, and wetlands. Fyke nets and trap nets are used in shallow
water while modified hoop nets and gill nets are used in deep or open waters
All passive sampling techniques should be checked and emptied 12 to 24 hours
after setting. Data collected by passive sampling techniques can be used to
determine relative abundance which are expressed as number/24 hours and weight
(kg)/24 hours (Ohio EPA, 1989).
59
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TABLE 4. AMOUNT OF 5% EMULSIFIABLE ROTENONE EQUIVALENT TO 0.5 PPM OR 1.0 PPM
PER ACRE-FEET OR POND OR LAKE TO BE SAMPLED
Rotenone (5% Emulsifiable) Application Rates
Acre-Feet Pints of 5% Rotenone
0.5 ppm 1.0 ppm
0.25
0.50
0.75
1.00
1.25
1.50
1.75
2.00
2.25
2.50
2.75
3.00
3.25
3.50
3.75
4.00
4,25
4.50
4.75
5.00
5.25
5.50
5.75
6.00
0.3
0.6
1.0
1.3
1.6
2.0
2.3
2.6
3.0
3.3
3.6
4.0
4.3
4.6
5.0
5.3
5.6
6.0
6.3
6.6
7.0
7.3
7.6
8.0
0.6
1.2
2.0
2.6
3.2
4.0
4.6
5.2
6.0
6.6
7.2
8.0
8.6
9.2
10.0
10.6
11.2
12.0
12.6
13.2
14.0
14.6
15.2
16.0
60
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TABLE 5. CUBIC CENTIMETERS (cc) OF LIQUID ROTENONE PER MINUTE FOR GALLONS OF
FLOW PER MINUTE
Flow of Stream
in Gallons
per Minute
10
20
30
40
50
60
70
80
90
100
200
300
400
500
Five Percent
in Cubi
0.5 ppm
0.019
0.038
0.057
0.076
0.095
0.114
0.132
0.151
0.170
0.189
0,379
0.568
0.757
0.946
(5%) Liquid Rotenone Requirements
c Centimeters Per Minute
1.00 ppm
0.038
0.076
0.114
0.151
0.189
0.227
0.265
0.303
0.341
0.379
0.757
1.136
1.514
1.893
1.5 ppm
0.057
0.114
0.170
0.227
0.284
0.341
0.397
0.454
0.511
0.568
1.136
1.703
2.271
2.839
2.0 ppm
0.076
0.151
0.227
0.303
0.379
0.454
0.530
0.606
0.681
0.757
1.514
2.271
3.028
3.785
61
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4.6.2 Entanglement nets
4.6.2.1 Gill and trammel nets are used extensively to sample fish populations
in estuaries, lakes, reservoirs, and larger rivers.
4.6.2.2 A gill net is usually set as an upright, vertical fence of netting
and can have either a variable or uniform mesh size. Experimental gill nets
made of monofilament may be 37.5 m long and constructed with 7.5 m panels of
15.2 mm, 22.9 mm, 25.4 mm, 40.6 mm, and 50.8 mm bar mesh, and the variable
mesh size gill nets are generally preferred. Fish attempt to swim through
the net and are caught in the mesh (Figure 8). Because the size of the mesh
determines the species and size of the fish to be caught, gill nets are
considered selective. The most versatile type is an experimental gill net
consisting of five different mesh size sections. Mesh sizes depend on the
size range of fish species to be sampled. A range of mesh sizes in an
experimental gill net is used to obtain samples of several year classes of a
single species, and it will also provide a greater chance to increase the
number of species caught. Gill nets made of multifilament or monofilament
nylon are recommended. Multifilament nets cost less and are easier to use,
but monofilament nets generally capture more fish. The floats and leads
usually supplied with the nets can cause net entanglement. To reduce this
problem replace the individual floats and float line with a float line made
with a core of expanded foam and use a lead-core leadline instead of
individual lead weights and lead line. Gill nets are usually set in open
waters to sample fishes in large rivers, lakes, and reservoirs. They can be
set at the surface, mid-depth, or on the bottom depending on the objectives of
the study and target species within the fish community. Gill nets should be
anchored and marked well in open water areas with floats on both ends.
4.6.2.3 The trammel net (Figure 9) has a layer of large mesh netting on each
side of loosely-hung, smaller gill netting. Small fish are captured in a
"bag" of the gill netting that is formed as the smaller-mesh gill netting is
pushed through an opening in the larger-mesh netting. Trammel nets are not
used as extensively as are gill nets in sampling fish.
4.6.2.4 Trammel nets can be fished in all types of habitats found in rivers
such as the Mississippi. If a backwater or quiet stretch of the river is to
be fished, the net is set. If the river channel is to be fished, the net is
floated or drifted downstream. Trammel nets are very efficient for taking
such fish as carp and buffalo. Trammel net float fishing is an excellent
method of sampling shovel nose sturgeon and freshwater drum.
4.6.2.5 Stationary gill and trammel nets are fished at right angles to
suspected fish movements (e.g., parallel to shore) and at any depth from the
surface to the bottom. They may be held in place by poles or anchors. The
anchoring method must hold the net in position against any unexpected water
movements such as, runoff, tides, or seiches.
4.6.2.6 Drifting gill or trammel nets are also set and fished the same as
stationary gear, except that they are not held in place but are allowed to
drift with the current. This method requires constant surveillance when
fishing. They are generally set for a short period of time. If currents are
62
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too great, stationary gear may be used, but heavy current can cause the net to
collapse.
4.6.2.7 Results for both trammel and gill nets are expressed as the number or
weight of fish taken per length of net per day (=catch per unit effort).
4.6.2.8 The use of gill nets in estuaries may present special problems, and
consideration should be given to tidal currents, predation, optimum fishing
time, and types of anchors, floats, and line. When gill net fishing in tidal
waters, it is recommended that reversing anchors be used for anchoring if the
nets are to be left unattended. Mushroom anchors and concrete blocks will not
hold down the nets during tidal cycles and may allow them to move considerable
distances if a high tidal cycle is present. The gill nets should be monitored
frequently and usually after a tidal cycle change as marine species usually
will not survive too long in gill nets. Dead fish tend to attract crabs which
tangle in the nets making them difficult to remove, When nets are set in the
mouths of creekSs the outgoing tidal cycle generally will be more productive.
4.6.2.9 In freshwater, monofilament gill nets are very effective for lake
herring, trout, lake whitefish, yellow perch, walleyes, and northern pike.
4.6.2,10 Necessary equipment for netting includes a pair of "clipper" pliers
for removing sharp pectoral and dorsal spines on catfish and bullheads when
these fish become tangled in the netting. Also, the gunnels of any boat used
in a net fishing operation should be free of rivets, cleats, etc. on which the
net can snag.
Figure 8. Gill net. Modified from Dumont and Sundstrom (1961),
4.6.3 Entrapment Devices
4.6.3.1 With entrapment devices, the fish enter an enclosed area (which may
be baited) through a series of one or more funnels and prevent excapement.
They are used to sample reserviors and wide river channels with slow velocity
conditions. Entrapment nets are set in structurally complex areas where fish
movement and density are anticipated to be highest in order to maximize net
catches.
63
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4.6.3.2 The hoop nets (modified hoop nets) and trap nets are the most common
types of entrapment devices used in fishery surveys. These traps are small
enough to be deployed from a small open boat and are relatively simple to set.
They are held in place with anchors or poles and are used in water deep enough
to cover the nets, or to a depth up to 4 meters.
4.6.3.3 The hoop net (Figure 10) is constructed by covering hoops or frames
with netting. It has one or more internal funnels and does not have wings or
a lead. The first two sections can be made square to prevent the net from
rolling in the currents.
4.6.3.4 The fyke net (Figure 11) is a hoop net with wings, or a lead, or both
attached to the first frame. The second and third frames can each hold funnel
throats, which prevent fish from escaping as they enter each section. The
opposite (closed) end of the net may be tied with a slip cord to facilitate
fish removal.
4.6.3.5 Hoop nets are fished in rivers and other waters where fish move in
predictable directions, whereas the fyke net is used when fish movement is
more random such as in lakes, impoundments, and estuaries. Hoop and fyke nets
can be obtained with hoops from 2 to 6 feet (0.6 to 1.8 meters) in diameter,
but any net over 4 feet (1.2 meters) in diameter is too large to be used in a
fishery survey.
4.6.3.6 Trap nets use the same principle as hoop nets for capturing fish, but
their construction is more complex. Floats and weights instead of hoops give
the net its shape. The devices are expensive, require considerable
experience, and are usually fished in waters deep enough to cover them.
4.6.3.7 One of the traps which has proven to be quite effective is a 3 x 6
foot frame with a 3 x 50 foot lead consisting of 1/2 inch square mesh of #126
knotless nylon. Traps with 1/4 inch mesh netting have also been used. Trap
nets are set with the lead perpendicular to the shoreline. They usually are
most effective in depths less than 25 feet with a minimum depth of about 3
feet.
4.6.3.8 One of the most simple types is the minnow trap, usually made of wire
mesh or glass, with a single inverted funnel. The bait is suspended in a
porous bag. A modification of this type is the slat trap (Figure 12); this
employs long wooden slats in a cylindrical trap, and when baited with cheese
bait, cottonseed cake, etc., is usually very successfully in sampling catfish
in large rivers.
4.6.3.9 Most fish can be sampled by setting trap and hoop nets of varying
sizes hi a variety of habitats. Hoop and trap nets are made of cotton or
nylon, but nets made of nylon have a longer life and are lighter when wet.
Protect cotton and nylon nets from decay by using the same methods of
treatment mentioned for seines in Subsection 4.2.1.5. The catch is recorded
as numbers or weight per unit of effort, usually fish per net day.
64
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Figure 9. Trammel Net. Modified from Dumont and Sundstrom (1961).
Figure 10. Hoop Nets. Modified from Dumont and Sundstrom (1961)
65
-------
Figure 11. Fyke Net. Modified from Dumont and Sundstrom (1961).
Figure 12. Slat Trap. Modified from Dumont and Sundstrom (1961).
66
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4.7 Pop Nets
4.7.1 Pop nets are useful for sampling fish in shallow riverine waters in
heavily vegetated and nonvegetated areas where seining or electroshocking may
be difficult (Larson et al., 1986; Dewey et al., 1989). Pop nets are set and
retrieved by two individuals and are easily dissembled for easy transport,
4.7.2 Pop nets (Figure 13) are rectangular devices, constructed of lightly
tarred 6.4 mm mesh netting. They are 1.8 m wide x 3.1 m long x 1.8 m high
when released and enclose an area of 6.5 m2. The top of the net is attached
to a rectangular polyvinylchloride frame filled with foam. The top of the
frame should be painted black to reduce the effect of color on fish avoidance
or attraction. They are designed to be set from the surface and released
with a mechanical device.
4.7.3 The pop net used in nonvegetated areas can simply be a rectangular
holding net with its top attached to the buoyant frame and its bottom panel
attached to a frame, 19 mm diameter galvanized pipe. After the pop net is
tripped, the net and attached frames are picked up and carried to shore as a
unit,
4.7.4 The enclosed bottom design pop net cannot be used in vegetated areas.
A pop net used in vegetated areas is constructed with an open-bottom. Its
bottom is split down the center and attached only along the two long sides of
the holding net. The bottom frame is still present but used only to hold the
buoyant top frame in position during the setting process and is not attached
Figure 13, Pop net. A. model for nonvegetative site after release.
set for release in vegetated site, show pipes used for
(*) and position of release mechanism (arrow).
mechanism. From Dewey et al. (1989).
Pop
net
closure
Release
B. Pop
bottom
C.
67
-------
at all to the holding net. Galvanized pipes 3.7 m long, are attached to each
of the two split sections and are used to hold the bottom sections to the
sides as the pop net is placed over vegetation (Figure 13,B). After the pop
net is released, these pipes are used to purse the bottom sections together
and thus enclose the catch. The sample is retrieved by carrying the top
frame, attached net, and pursing pipes to shore as a unit. The bottom frame
is retrieved separately.
4.7.5 Pop nets have two release mechanisms (Figure 13,B and C), each
consisting of two devices at opposite sides of the net. At each position, a
piece of aluminum flat bar, attached to the top frame, fits into a slot in the
bottom frame. An L-shaped extension attached to the trip rod fits through
matching holes in the flat bar and in the bottom frame to hold the top and
bottom frames together. The trip rods for both mechanisms are joined by a
lead core line, to which is tied a 5 cm trip cord. When the trip cord is
pulled, both trip rods release simultaneously, allowing the buoyant top frame
to rise. In the set position, these release mechanisms hold the upper and
lower frames together in a low profile (9.5 cm high), which increases
stability in currents.
4.8 Miscellaneous Fish Methods
4.8.1 Underwater Methods
4.8.1.1 Direct observation techniques can be used to study the structure of
fish assemblages, spawning, feeding, and movement, etc. For techniques on
direct underwater observation which involve the use of divers (snorkeling and
scuba) to study fish populations, see Helfman (1983) and Pearsons et al.
(1992).
4.8.2 Hydroacoustic Techniques
4.8.2.1 Hydroacoustic assessment techniques are generally applied to methods
which use equipment such as sonars or depthsounders. The hydroacoustic
techniques use sound from these devices that are actively transmitted and
information extracted from the returning echoes to detect fish and make
qualitative and quantitative estimates of biomass. For a review, discussion,
and guidelines of fishery hydroacoustics, see Thorne (1983).
4.8.2.2 Information on hydroacoustic equipment for fisheries evaluations can
be obtained from the following company:
Hydroacoustic Technology, Inc.
715 NE North!ake Way
Seattle, WA 98105, Telephone (206) 633-3383.
4.8.3 Underwater Biotelemetry
4.8.3.1 These techniques are often used to monitor the locations, behavior,
and physiology of free-ranging fish, and involves attaching a device that
relays biological information. For a review and discussion of telemetry
methods, see Winter (1983).
68
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4.8.3.2 Information on a field proven digitally encoded radio telemetry
system for fisheries evaluations can be obtained from the following company:
Lotek Engineering, Inc.
115 Pony Drive
Newmarket, Ontario, Canada L3Y 7B5
Telephone (416)
4.9 Literature Cited
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influencing the collection efficiency of estuarine fishes. Trans, Amer.
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(18th edition). American Public Health Association, Washington, DC.
Armour, C.L., K.P. Burnham, and W.S. Platts. 1983. Field methods and
statistical analyses for monitoring small salmonid streams. U.S.
Department Interior, Fish and Wildlife Service, Washington, DC. 20240.
FWS/OBS-83/33.
ASTM. 1992. Classification for fish sampling. Designation: D 4211-82
(Reapproved 1987). Annual Book of ASTM Standards, Section 11, Volume
11.04, American Society for Testing and Materials, Philadelphia, PA. pp
59-60.
Bayley, P.B., R.W. Larimore, and D.C. Dowling. 1989. Electric seine as a
fish-sampling gear in streams. Trans. Amer. Fish. Soc. 118:447-453.
Blair, A.A. 1958. Back-pack shocker. Can. Fish Cult. 23:33-37.
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small streams. Trans. Am. Fish. Soc. 92:307-310.
Bone, J.N. 1970. A method for dispensing rotenone emulsions. British
Columbia Fish and Wildlife Branch, Fish Management Report 62:1-3.
Bowers, C.C. 1955. Selective poisoning of gizzard shad with Rotenone, Prog.
Fish-Cult. 17(3):134-135.
Braem, R.A. and W.J. Ebel. 1961. A back-pack shocker for collecting lamprey
ammocoetes. Prog. Fish-Cult. 23:87-91.
Carter, E.R. 1954. An evaluation of nine types of commercial fishing gear in
Kentucky Lake. Trans. Kentucky Academy Science 15:56-80.
Cohen, J.M., Q.H. Pickering, R.L. Woodward, and W. Van Heruveleln. 1960. The
effect of fish poisons on water supplies. J. Am. Water Works Assoc.
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Cohen, J.M., Q.H. Pickering, R.L. Woodward, and W. Van Heruveleln. 1961. The
effect of fish poisons on water supplies. J. Am. Water Works Assoc.
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Cowx, I. G. (ed.). 1990. Developments in electric fishing. Blackwell
Scientific Publ., Cambridge, MA. (available from the Amer. Fish. Soc.,
Bethesda, MD).
Cowx, I.G. and P. Lamarque (eds,). 1990. Fishing with electricity.
Blackwell Scientific Publ., Cambridge, MA. (available from the
Amer.Fish. Soc., Bethesda, MD).
Cross, D. G. and B. Stott. 1975. The effects of electric fishing on the
subsequent capture of fish. J. Fisheries Biol. 7(3):349-357.
Cyr, H., J.A. Downing, S. Lalonde, S.B. Baines, and M.L. Pace. 1992.
Sampling larval fish populations: Choice of sample number and size.
Trans. Amer Fish. Soc. 121:356-368.
Dauble, D.D. and R.H. Gray. 1980. Comparison of a small seine and a backpack
electroshocker to evaluate near shore fish populations in rivers. Prog.
Fish-Cult. 42:93-95.
Davies, W.D. and W.L. Shelton. 1983. Sampling with toxicants. In: Nielsen,
L.A. and D.L. Johnson (eds.). Fisheries Techniques. American Fisheries
Society, Bethesda, MD. pp. 199-213.
Dewey, M.R., I.E. Holland-Bartels, and S.T. Zigler. 1989. Comparison of fish
catches with buoyant pop nets and seines in vegetated and nonvegetated
habitats. N. Amer. J. Fish. Manage. 9:249-253.
Dove!, W.L. 1964. An approach to sampling estuarine macroplankton.
Chesapeake Sci. 5:77-90.
Dumont, W.H. and G.T. Sundstrom. 1961. Commercial fishing gear of the United
States. U.S. Fish and Wildlife Circular No. 109, U.S. Government
Printing Office, Washington, DC. 61 pp.
Elson, P.F. 1950. Usefulness of electrofishing methods. Can. Fish Cult.
9:3-12.
Everhart, W.H., A.W. Eipper, W.D. Youngs. 1975. Principles of fisheries
science. Cornell University Press, Ithaca, N.Y. 288 pp.
Fisher, K.C. 1950. Physiological considerations involved in electrical
methods of fishing. Can. Fish Cult. 9:26-34.
Frankenberger, L. 1960. Application of a boat-rigged direct-current shocker
on lakes and streams in west-central Wisconsin. Prog. Fish-Cult
22(3):124-128.
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Friedman, R. 1974. Electrofishing for population sampling. A selected
bibliography. Res. Serv. Br., Office Library Serv., U.S. Dept.
Interior, Biographical Series 31, 15 pp.
Funk, J.L. 1947. Wider application of electrical fishing method of
collecting fish. Trans. Amer. Fish. Soc. 77:49-64.
Gammon, J.R. 1973. The effect of thermal inputs on the populations of fish
and macroinvertebrates in the Wabash River. Purdue Univ. Water
Resources Res. Cen. Tech. Rep. 32. 106 pp.
Gammon, J.R. 1976. The fish populations of the middle 340 km of the Wabash
River. Purdue Univ. Water Resources. Res. Cen. Tech. Rep. 86. 73 pp.
Gulland, J.A. 1980. General concepts of sampling fish. Pages 7-12. In: T.
Backiel and R.L. Welcomme (eds.). Guidelines for sampling fish in
inland waters. European Inland Fisheries Advisory comm. Tech. Pap. 33.
Hartley, W.G. 1980. The use of electrical fishing for estimating stocks of
freshwater fish. Pages 91-95. In: T. Backiel and R.L. Welcomme
(eds.). Guidelines for sampling fish in inland waters. European Inland
Fish. Advisory Comm. Tech. Pap. 33.
Haskell, D.C. and W.F. Adelman, Jr. 1955. Effects of rapid direct current
pulsations on fish. New York Fish Game J. 2(1):95-105.
Haskell, D.C., D. Geduldiz, and E. Snolk. 1955. An electric trawl. New York
Fish Game J. 2(1):120-125.
Hayes, M.L. 1983. Active fish capture methods. In: Nielsen, L.A. and D.L.
Johnson (eds.). Fisheries techniques. American Fisheries Society,
Bethesda, MD. pp. 123-145
Helfman, G.S. 1983. Underwater methods. In: Nielsen, LA. and D.L.
Johnson (eds.). Fisheries Techniques. American Fisheries Society,
Bethesda, MD. pp. 349-369.
Henderson, H.F. 1980. Some statistical considerations in relation to
sampling populations of fishes. Pages 167-176. In: T. Backiel and
R.L. Welcomme (eds.). Guidelines for sampling fish in inland waters.
European Inland Fish. Advisory Comm. Tech. Pap. 33.
Hendricks, M.L., C.H. Hocutt, Jr., and J.R. Stauffer, Jr. 1980. Monitoring
of fish in lotic habitats. Pages 205-231. In: C.H. Hocutt, Jr. and
J.R. Stauffer, Jr. (eds.). Biological monitoring of fish. Lexington
Books, Lexington, MA.
Hocutt, C.H., P.S. Hambrick, and M.T. Masnik. 1973. Rotenone methods in a
large river system. Arch. Hydrobiology 72(2):245-252.
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Holden, A.V, 1980. Chemical methods. Pages 97-104. In: T. Backiel and
R,L. Welcomme (eds,). Guidelines for sampling fish in inland waters.
European Inland fish. Advisory Comm. Tech. Pap. 33.
Holten, G.D, 1954. West Virginia's electrical fish collecting methods.
Prog. Fish-Cult. 16:10-18.
Hooper, F. 1960. Pollution control by chemicals and some resulting problems.
Trans. Second Seminar Biol. Problems in Water Pollution, April 20-24,
USPHS, Robert A. Taft San. Engr. Ctr., Cincinnati, pp. 241-246.
Hubert, W.A. 1983. Passive capture techniques. In: Nielsen, L.A. and D.L.
Johnson (eds.). Fisheries techniques. American Fisheries Society,
Bethesda, MD. pp. 95-122.
Hughes, R.M., D.P. Larsen. 1988. Ecoregions: An appraoch to surface water
protection. J. Water Pollut. Control Fed. 60:486-493.
Hughes, R.M., D.P. Larsen, and J.M. Omernik. 1986. Regional reference sites:
A method for assessing stream potentials. Environ. Manage. 10:629-635.
Hughes, R.M., E. Rexstad, And. C.E. Bond. 1987. The relationships of aquatic
ecoregions, river basins, and physiographic provinces to the
ichthyogeographic regions of Oregon. Copeia 1987:423-432.
Hunt, R.L. 1992. Evaluation of trout habitat improvement structures in three
high-gradient streams in Wisconsin. Technical Bulletin No. 179.
Department of Natural Resources, Box 7921, Madison, WI
Johnson, D.L. and L.A. Nielsen 1983. Sampling considerations. In: Nielsen,
L.A. and D.L. Johnson (eds.). Fisheries techniques. American Fisheries
Society, Bethesda, MD. pp. 1-21.
Kolz, A.L. 1989. A power transfer theory for electrofishing. Fish and
Wildlife Technical Report 22, Fish and Wildlife Service, U.S. Department
of the Interior, Washington, DC. pp. 1-11.
Kolz, A.L. and J.B. Reynolds. 1989a. Electrofishing, a power related
phenomenon. Fish and Wildlife Technical Report 22, Fish and Wildlife
Service, U.S. Department of the Interior, Washington, DC.
Kolz, A.L. and J.B. Reynolds. 1989b. Determination of power threshold
response curves. Fish and Wildlife Technical Report 22, Fish and
Wildlife Service, U.S. Department of the Interior, Washington, DC. pp.
15-25.
Krumholz, L.A. 1948. The use of Rotenone in fisheries research. J. Wildl.
Mgmt. 12(3):305-317.
Lagler, K.F. 1956. Freshwater fishery biology. Second Edition. William C.
Brown Co., Dubuque, Iowa. 421 pp.
72
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Lagler, K.F. 1978. Capture, sampling and examination of fishes. Pages 7-
47. In: Methods for assessment of fish production in freshwater.
Blackwell Sci. Publ., Oxford. IBP No. 3.
Larimore, R.W. 1961. Fish population and electrofishing success in a warm
water stream. J, Wild!. Mgmt. 25(1):1-12.
Larimore, R.W., L. Durham, and G.W. Bennett. 1950, A modification of the
electric fish shocker for lake work, J. Wild!. Mgmt. 14(3}:320-323.
Larson, F.W., D.L, Johnson, and W.E. Lynch, Jr. 1986. A buoyant pop net
for accurately sampling fish at artificial habitat structures. Trans.
Amer. Fish. Soc. 115:351-355.
Latta, W.C. and G.F. Meyers. 1961. Night use of DC electric shocker to
collect trout in lakes. Trans. Amer, Fish. Soc, 90(l):81-83.
Lawrence, J.M. 1955. Preliminary results on the use of potassium
permanganate to counteract the efffects of Rotenone on fish. Proc.
Southeastern Asso, Game and Fish 'Comm., October 2-5, 1955, pp. 1-13.
Lawrence, J.M. 1956. Preliminary results of the use of KMN04 to counteract
the effects of rotenone. Proj. Fish-Cult. 10(1):15-21.
Lennon, R.E. 1959. The electrical resistivity in fishing investigations.
U.S. Fish Wildl. Serv., Spec. Sci. Rept. Fish No. 287:1-13.
Lennon, R.E. 1961. A fly-rod electrode system for electrofishing. Prog.
Fish-Cult. 23{2):92-93.
Lennon, R.E. and P.S. Parker. 1958. Application of salt in electro-fishing.
U.S. Fish Wildl. Serv., Spec. Sci. Rept. Fish 280, 11 pp.
Lennon, R.E., J.B. Hunn, R.A. Schnicks and R.M. Burress. 1971. Reclamation
of ponds, lakes, and streams with fish toxicant~-A review: U.S. Fish
and Wildlife Service, FishTechnical report 100, 9 pp.
Loeb, H.A. 1955. An electrical surface device for crop control and fish
collection in lakes. New York Fish Game J. 2:220-221.
Mackenthun, K.M. 1969. The practice of water pollution biology. USDI,
FWPCA, 281 pp.
Mackenthun, K.M. and W.M. Ingram. 1967. Biological associated problems in
freshwater environments, their identification, investigation and
control. USDI, FWPCA, 287 pp.
Mansuetti, A.J. and J.D. Hardy, Jr. 1967. Development of the fishes of the
Chesapeake Bay Region, An atlas of egg, larval, and juvenile stages Part
1. Nat. Res. Insti. Univ. Maryland, Baltimore, MD. 202 pp.
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Marking, L.L. 1992. Evaluation of toxicants for the control of carp and
other nuisance fishes. Fisheries 17:6-13.
Massman, W.H., E.G. Ladd, and H.N. McCutcheon. 1952. A surface trawl for
sampling young fished in tidal rivers. Trans. North Amer. Wildl. Conf.
17:386-392.
McCrimmon, H.R. and A.M. Berst. 1963. A portable AC-DC backpack fish shocker
designed for operation in Ontario streams. Prog. Fish-Cult. 25(3):159-
162.
Meyer, P.P., R.A. Schnick, K.B. Cumming, and B.L. Berger. 1976. Registration
status of fishery chemicals, February 1976. Preg. Fish-Cult. 38(1):3-7.
Nester, R.T. 1987. Horizontal ichthyoplankton tow-net system with
unobstructed net opening. North Amer. J. Fisheries Mgmt. 7:148-150.
Nester, R.T. 1992. Great Lakes Fishery Laboratory, Fish & Wildlife Service,
Ann Arbor, MI. Personal Communication.
Novotny, D.W. and G.R. Priegel. 1971. A guideline for portable direct current
eleetrofishing system. Tech. Bull. No. 51, Wis. Dept. Nat. Res., 22 pp.
Novotny, D.W. and G.R. Priegel. 1974. Electrofishing boats improved designs
and operational guidelines to increase the effectiveness of boom
shockers. Tech. Bull. No. 73. Wis. Dept. Nat., 48 pp.
0'Gorman, R. 1984. Catches of larval rainbow smelt Osmerus mordax and
alewife Alosa pseudoharengus in plankton nets of different mesh sizes.
J. Great Lakes Res. 10:73-77.
Ohio EPA. 1987a. Biological criteria for the protection of aquatic life:
Volume I. The role of biological data in water quality assessment.
Division of Water Quality Monitoring and Asssessment, Surface Water
Section, Columbus, OH.
Ohio EPA. 1987b. Biological criteria for the protection of aquatic life:
Volume II. Users manual for biological field assessment of Ohio surface
waters. Division of Water Quality Monitoring and Asssessment, Surface
Water Section, Columbus, OH.
Ohio EPA. 1989. Biological criteria for the protection of aquatic life:
volume III. Standardized field and laboratory methods for assessing
fish and macroinvertebrate communities. Divsion of Water Quality
Monitoring and Assessment, Surface Water Section, Columbus, OH.
Ohio EPA. 1990, Fish evaluation group safety manual. Ohio Environmental
Protection Agency, ecological Assessment Section, Division of Water
Quality Planning and Assessment. Columbus, OH.
Omernik, J.M. 1987. Ecoregions of the conterminous United States. Ann. Ass.
Am. Geo. 77:117-125.
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Omernik, J.M. and A.L. Gallant. 1988. Ecoregions of the upper midwest
states. U.S. Environmental Protection Agency, Environmental Research
Laboratory, Corvallis, OR.
Orth, Donald J. 1983. Aquatic habitat measurements. In: Nielsen, L.A. and
D.L. Johnson (eds.). Fisheries techniques. American Fisheries
Society, Bethesda, MD, pp. 61-84.
Pearsons, T.N., H.W. Li, G.A. Lamberti. 1992. Influence of habitat
complexity on resistance to flooding and resilience of stream fish
assemblages. Trans. Amer. Fish. Soc. 121:427-436.
Plafkin, J.L., M.T. Barbour, K.D, Porter, S.K. Gross, and R.M. Hughes. 1989.
Rapid bioassessment protocols for use in streams and rivers: benthic
macroinvertebrates and fish. EPA/440/4-89/001. U.S. Environmental
Protection Agency, Assessment and Watershed Protection Division,
Washington, DC.
Platts, W.S., W.F. Megahan, and G.W. Minshall. 1983. Methods for evaluating
stream riparian and biotic conditions. U.S. Forest Serv. Forest Range
Exp. Stn., Gen. Tech. Eep.. INT-138.
Pratt, V.S. 1951. A measure of the efficiency of alternating and direct
current fish shockers. Trans. Amer. Fish. Soc. 81(l):63-68.
Price, R.W. and J. B. Haus. 1963. Aids for stream reclamation. Prog.
Fish. Cult. 25(l):37-39.
Rankin, E.T. 1989. The qualitative habitat evaluation index (QHEI):
rationale, methods, and application. Ecological Assessment Section,
Division of Water Quality Planning & Assessment, P.O. Box 1049, 1800
WaterMark Drive, Columbus, OH.
Reynolds, J.B. 1983. Electrofishing. In: Nielsen, LA. and D.L.
Johnson (eds.). Fisheries Techniques. American Fisheries Society,
Bethesda, MD. pp. 147-163.
Ricker, W.E. (ed.). 1971. Methods for assessment of fish production in fresh
waters. Oxford and Edinburgh, Blackwell Scientific Publication,
International Biological Programme Handbook 3, 384 pp.
Rollefson, M.D. 1958. The development and evaluation of interrupted direct
current electrofishing equipment. WY Game Fish Dept. Coop. Proj. No. 1,
123 pp.
Rollefson, M.D. 1961. The development of improved electrofishing equipment.
In: Proc. 41st. Ann. Cong. West. Assoc. St. Game and Fish Comm., pp
218-228.
Rounsefell, G.A. and W.H. Everhart. 1953. Fishery science: Its methods and
applications. John Wiley and Sons, New York. pp. 444.
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Sanders, R.E. 1991. A 1990 night electrofishing survey of the upper Ohio
River mainstem (RM 40.5 to 270.8) and recommendations for a long-term
monitoring program. Ohio Dept. Nat. Res. (ODNR), Division of Wildlife,
1840 Belcher Dr., Columbus, OH.
Sanders, R.E. 1992. Day versus night electrofishing catches from near-shore
waters of the Ohio and Muskingum Rivers. Ohio J. Sci. 92(3):In Press.
Schnick, R.A. 1974. A review of the literature on the use of rotenone in
fisheries. La Crosse, Wis., Fish Control Laboratory, 130 pp.
(Available from U.S. Dept. Commerce, Nat. Tech. Information Serv (NTIS).
Springfield, Va 22161 as publication FWS-0-74 15.)
Schnick, R.A. and P.P. Meyer. 1978. Registration of thirty-three fishery
chemicals: Status of research and estimated costs of required contract
studies. United States Fish and Wildlife Service, Investigations in
Fish Control No. 86:1-19, Washington, District of Columbia, USA.
Schreck, C.B. and P.B. Moyle (eds.). 1990. Methods for fish biology. Amer.
Fish. Soc., Bethesda, MD
Seehorn, M.E. 1968. An inexpensive backpack shocker for one man use. In:
Proc. 21st. Ann. Cong. Southeastern Assoc, Game and Fish Comm., pp. 516-
524.
Sharpe, F. P. 1964. An electrofishing boat with a variable-voltage pulsator
for lake and reservoir studies. U.S. Bureau Sport Fisheries and
Wildlife Circular 195. 6 pp.
Sharpe, F.P. and W.T. Burkhard. 1969. A lightweight backpack high voltage
electrofishing suit. U.S. Bur. Sport Fisheries and Wildlife Res. Publ.
78, 8 pp.
Simon, T.P. 1989. Rationale for a family-level ichthyoplankton index for
use in evaluating water quality. In: W.S. Davis and T.P. Simon (eds.).
Proceedings of the 1989 Midwest Pollution control biologists meeting,
Chicago, Illinois. U.S. Environmental Protection Agency, Chicago, IL.
pp. 41-65.
Smith, P.E. and S.L. Richardson. 1977. Standard techniques for pelagic fish
egg and larva studies. Food and Agriculture Organization of the United
Nations, Fisheries Technical Paper 175, Rome, Italy.
Snyder, D.E. 1983. Fish eggs and larvae. In: Nielsen, L.A. and D.L.
Johnson (eds.). Fisheries techniques. American Fisheries Society,
Bethesda, MD. pp. 165-197.
Starrett, W.C. and JP.G. Barnickol. 1955. Efficiency and selectivity of
commercial fishing devices used on the Mississippi River. Illinois
Natural History Survey Bulletin 26:325-366.
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Stubbs, J.M. 1966. Electrofishing, using a boat as the negative. In: Proc.
19th Ann. Conf, Southeastern Assoc, and Fish Comm,, pp. 236-245,
Thome, R.E. 1983. Hydroacousrics, In: Nielsen, LA, and D.L. Johnson
(eds.). Fisheries Techniques. American Fisheries Society, Bethesda,
MD. pp. 239-259.
Tranter, D.J. and P.E. Smith. 1968. Filtration performance. Monographs on
Oceanographic Hethodology 2:27-56.
Trent, W.L, 1967. Attachment of hydrofoils to otter boards for taking surface
samples of juvenile fish and shrimp. Ches. Sci. 8(2):130-133,
USEPA 1978. Quality assurance guidelines for biological testing. EPA-600/4-
78-043. U.S. Environmental Protection Agency, Environmental Monitoring
and Support Lab., Las Vegas, NV
U.S. Fish and Wildlife Service. 1991. Principles and techniques of
electrofishing. Fisheries Academy, U.S. fish and Wildlife Service,
Office of Technical Fisheries Training, Kearneysville, WV.
Vincent, R. 1971. River electrofishing and fish population estimates. Prog.
Fish-Cult. 33(3):163-169.
Winter, J.D. 1983. Underwater biotelemetry. In: Nielsen, L.A. and D.L,
Johnson (eds.). Fisheries techniques. American Fisheries Society,
Bethesda, MD. pp. 371-395.
Yeh, C.F. 1977. Relative selectivity of fishing gear used in a large
reservoir in Texas. Trans. Am. fish. Soc. 106:309-313.
Zippin, C. 1956. An evaluation of the removal method of estimating animal
populations. Biometrics 12:163-169.
Zippin, C. 1958. The removal methods of population estimation. J. Wildl.
Manage. 22:82-90.
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SECTION 5
FISH PROCESSING
5.1 Introduction
5.1.1 After fish are collected, they must be either examined and identified
in the field or if voucher specimens are required, they must be fixed
immediately for subsequent identification in the laboratory. If the sampling
crew have difficulty identifying any specimens in the field, those specimens
must be fixed and later identified in the laboratory. The decision to
preserve specimens should depend on study objectives. One set of specimens
should be preserved during the study (especially in the early stages) so that
a vouchered, archived reference collection of each species from different
study areas or eeoregions will be available to investigators. The study team
should be become familiar with characteristics of the specimens difficult to
identify. For general purposes, formalin is usually used as a fixing agent
(ASIH, 1988). This fixative solution helps retain chromatophore patterns
which aid in species identification. When using formalin, care must be taken
because it is highly allergenic, toxic, and dangerous to human health
(carcinogenic) if used improperly.
5.1.2 If specimens are to be kept alive, they should be placed in a live
well, container, or bucket and processed upon completion of sampling at each
site or when the live well container or bucket are full. To minimize fish
mortality in the live well or bucket, water should be changed periodically or
aerated with a battery-powered pump. Fish should be handled carefully and
released immediately after they are identified to species, examined for
external anomalies, and weighed if necessary. Every effort should be made to
minimize fish handling and holding times.
5.1.2.1 If a large number of the fish specimens are to be kept alive for
later study, see Stickney (1983) for a discussion and guidelines on caring for
and handling live fish.
5.2 Fixation and/or Preservation of Fish Samples
5.2.1 Fixation is the process of rapidly killing and chemically stabilizing
fish tissues to maintain anatomical form and structure. Preservation is the
process by which fixed tissues are maintained in that condition for an
indefinite period of time,
5.2.2 Fish and ichtyoplankton should be fixed and preserved (Table 1) in the
field in neutral buffered 10% formalin or borax buffered 10% formalin (a 9:1
ambient water dilution of 100% formalin) for 24 hours or longer, depending on
size of fish (Haedrich, 1983, Lagler, 1956, Lagler et al., 1962, Humason,
1974, and Knudsen, 1966). The sodium phosphate monobasic and sodium phosphate
dibasic, or borax, acts as a buffer which neutralizes the acidic effect of the
formaldehyde. This mixture retards shrinkage in fish, prevents the hardening
of soft body parts, and prevents decalcification of the tissues (Lagler et
al., 1962). Fish should remain in the formalin solution for at least 1-2
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weeks to fix the tissue. Fixation may take from a few days with small
specimens to a week or more with large forms. Large fish or containers with
closely packed fish or temperatures greater than 26.7°C (80°F) require a
stronger solution of one part formalin to seven or eight parts water for
fixation. Stronger solutions of formalin can cause gaping or distortion of
the mouth and gills, thus care should be taken to obtain correct
concentrations when making up the formalin solution (Ohio EPA, 1989).
TABLE 1. FORMULATION OF FORMALIN FIXATIVE SOLUTION
37% formaldehyde (100% formalin) 100 mL
Distilled water 900 mL
and
Sodium phosphate monobasic (NaH?P04 • H20) 4 g
Sodium phosphate dibasic (Na2HP04) 6.5 g
or
Add one teaspoon of borax per 1/2 gallon of the formalin
5.2.3 Since the volume of collected fishes must be taken into account upon
fixation, formalin for field use should be stronger than 10%, and even 20%
will not hurt. Formaldehyde gas reaches saturation in water at about 37% by
weight; this saturated solution is called 100% formalin. Isopropyl alcohol
and ethyl alcohol are preservatives, not fixatives. These preservatives do
not fix the tissues, a necessary procedure for tissue preparation, staining,
etc.
5.2.4 After fixation in the formalin, some scientists transfer the specimens
to a preservative for storage. Ethyl alcohol (70-75%) or isopropanol (40-45%)
preservation keeps specimens more pliable than formalin and makes working with
them easier. Specimens should be rinsed in water to wash off any excess
formalin, placed in a 35% alcohol wash for 2-3 weeks, switched to a 50%
alcohol wash for 2-3 weeks, and placed in a 70%-75% aqueous solution of ethyl
alcohol or 40-45% isopropanol alcohol for permanent preservation and storage
(Haedrich, 1983; Ohio EPA, 1989). Fish should be stored in glass or plastic
containers or stainless steel vats for large specimens. Metal containers
should not be used. It is important that the containers be tightly sealed to
prevent evaporation of the preservative.
5.2.5 Specimens are kept in tightly sealed museum jars, along with their
field data. The preservatives will always modify the color, and light will
further bleach the fish specimens so the various markings and colors of fish
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should be documented if the specimens are to be identified later. It is
advisable to store specimens in the dark at 18°C to minimize evaporation and
bleaching.
5.2.6 Specimens larger than 7,5 cm should be slit on the side at least one-
third of the length of the body cavity or injected with a hypodermic syringe
to permit the preservative to reach the internal organs. Large and heavy
fish (1-2 pounds) should also be injected in the muscles on each side of the
backbone with formalin. Fish should be slit on the right side, because the
left side is generally used for measurement, scale sampling and photographic
records.
5,2,7 Samples for fish tissue contaminant analysis or electrophoresis must be
iced, placed in dry ice, or liquid N, for temporary storage or shipping. Fish
samples for pesticide analysis shoula be wrapped in aluminum foil, see Section
10, Guidelines for Fish Sampling and Tissue Preparation for Bioaccumulation
Contaminants, and placed in a cooler with ice. The sample must be frozen as
soon as possible after collection. Fish collected for metals analysis should
be placed in plastic bags. All samples should be doubled tagged, with one tag
attached outside the foil or plastic bag and one tag inside.
5.2.8 Special preservation techniques must be used for histological,
histochemical, or biomarker analyses, and the investigator should be aware of
such techniques before collecting tissue samples (Humason, 1974).
5.3 Labelling of Specimens In Field and Laboratory
5.3.1 Each specimen or specimens from a collecting site should be carefully
labelled with at least the information asked for in the examples of labels in
Figure 1.
5.3.1.1 Collection information should be both on and in the container, a tag,
or a paper label, If paper labels are used, they should be made of 100% rag
(waterproof) and labelled with India ink or a No. 2 soft lead pencil.
5.4 Species Identification
5.4.1 Many fish can be field identified with certainty. However, the
following procedures for fish identification and verification of difficult
specimens are recommended by Lowe-McConnell (1978):
1. Assemble and use the best available keys and checklists (see Section 8,
Fish Bioassessment Protocols for Use in Stream and Rivers, Subsection 8.14,
Selected References for Determining Fish Tolerance, Trophic, Reproductive, and
Origin Classifications and Section 12, Fisheries Bibliography, Subsection,
12.5 Fish Identification).
2, Key fish to species level,
3. Maintain a voucher collection in the laboratory for comparison of
specimens.
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4. Verify difficult species identifications with pictures, published
descriptions, known geographic range, museum and lab voucher specimens, or
have the specimen identified or verified by a specialist.
Project
Date
Location
FIELD SAMPLE DATA LABEL
Time
Collection No.
i County
Collector(s)
Type of sample
Method of collection
State/Country
Preservative(s).
A. Long Form
Date
FIELD SAMPLE DATA LABEL
Collection No.
Location
Collector(s)
Type of sample
Preservative(s)
B. Short Form
Figure 1. Examples of field sample data labels.
form.
A. Long form, B. Short
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5.4.2 Scientific nomenclature of all specimens should follow the
recommendations of the American Fisheries Society (Robins et a!., 1990).
5.4.4 Biomonitoring laboratories should maintain a fish reference collection.
Unique specimens should also added to the collection. The collection should
be archived in a computer data base which cross-references field data and
other pertinent information about the study.
5.5 Literature Cited
ASIH (American Society of Ichthyologists and Herpetologists), American
Fisheries Society, and American Institute of Fishery Research
Biologists. 1988. Guidelines for use of fishes in field research.
Fisheries (Bethesda) 132:16-23.
Haedrich, R.L. 1983. Reference collections and faunal surveys. In:
Nielson, L.A. and D.L. Johnson (eds.). Fisheries techniques. Amer.
Fish. Soc., Bethesda, MD. pp. 275-282.
Humason, Q.I. 1974. Animal tissue techniques. W.M. Freeman Co., San
Francisco, CA.
Knudsen, J.W. 1966. Biological Techniques. Harper and Row, Publishers, New
York, NY.
Lagler, K.F. 1956. Freshwater Fishery Biology. Wm. C. Brown Company
Publishers, Dubuque, IA.
Lagler, K.R., J.E. Bardach, and R.R. Miller. 1962. Ichthyology. John Wiley
& Sons, Inc., New York, NY.
Lowe-McConnell, R.H. 1978. Identification of freshwater fishes. Pages 48-
83. In: T. Bagenal (ed.). Methods for assessment of fish production in
fresh waters. IBP Handbook No. 3, Blackwell Sci, Publ., Oxford.
Ohio EPA. 1989. Biological criteria for the protection of aquatic life:
Volume III. Standardized biological field sampling and laboratory
methods for assessing fish and macroinvertebrate communities. Ohio
Environmental Protection Agency, Division Water Quality Monitoring and
Assessment, Ecological Assessment Section, Columbus, Ohio.
Robins, C.R., R.M. Bailey, C.E. Bond, J.R. Brooker, E.A. Lachner, R.N. Lea,
and W.B. Scott. 1990. Common and scientific names of fishes from the
United States and Canada. Amer. Fish. Soc., Special Publication 20.
Amer, Fish. Soc., Bethesda, MD.
Stickney, R.R. 1983. Care and handling of live fish. In: Nielson, L.A. and
D.L. Johnson (eds.). Fisheries techniques. Amer. Fish. Soc., Bethesda,
MD. pp. 85-94.
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SECTION 6
SAMPLE ANALYSIS TECHNIQUES
6.1 Introduction
6.1.1 One of the major concerns of USEPA, other federal, state and private
agencies or laboratories is to describe water quality and habitat quality in
terms which are easily understood by the nonbiologist. Fish studies
frequently include the number of specimens captured per unit area or unit
time. Also, the fish can be measured, weighted, aged, and sexed to provide
comparative data between populations in different habitats. The purpose of
this section is not to recommend one particular data evaluation method, but to
point out a number of more common methods. Some of these methods may not be
applicable to every stream, lake, or water body in the United States.
Methods, techniques, and biological criteria used to study fisheries biology
and to analyze fisheries data are described in this manual, elsewhere in
Bagenal (1978), Lager (1956, 1978), Carlander (1969), Everhart et al. (1975),
Gulland (1983), Nielsen and Johnson (1983), Schreck and Moyle (1990), USEPA
(1990, 1991), and also in other current literature. To supplement the
statistics and data evaluation methods in this section and for additional
biometrics, consult the statistical references listed in Section 1,
Introduction, Subsection 1.16.1. For other multivariate analyses and other
techniques to relate distribution to environmental variables and gradients,
confer with Matthews (1985), Matthews and Robison (1988), Mayden (1985; 1988),
and McAllister et al. (1986).
6.1.2 Water quality and habitat quality are reflected in the species
composition and diversity, population density and biomass, and physiological
condition of indigenous communities of aquatic organisms, including fish. A
number of data interpretation methods have been developed based on these
community characteristics to indicate the health and water quality of the
aquatic environment, the degree of habitat degradation, and also to simplify
communication problems regarding management decisions.
6.2 Data Recording
6.2.1 The sample records should include collection number, name of water
body, date, locality, names of sample collectors, and other pertinent
information associated with the sample. Make adequate field notes for each
collection. Use water-proof ink and paper to ensure a permanent record.
Place the label (Figure 1; also see Section 2, Quality Assurance and Quality
Control; Section 5, Fish Specimen Processing) inside the container with the
specimens only when fixing or preserving fish for physical examination (Note:
do not place the label with fish if they are to be chemically analyzed.) and
have the label bear the same number or designation as the field notes,
including the locality, date, and collector's name. Place a numbered tag on
the outside of the container to make it easier to find a particular
collection. Place any detailed observations about a collection on the field
data sheet (see Section 4, Sample Collection for Analysis of Structure and
Function of Fish Communities and Section 8, Fish Bioassessment Protocols for
83
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Use in Streams and Rivers for examples of field data sheets). Record fishery
catch data in standard units such as number or weight per area or unit of
effort. Use the metric system for length and weight measurements. Designate
any chemical analyses to be performed, e.g., toxaphene analysis.
6.3 Fish Identification
6.3.1 Proper identification of fish to species level is mandatory in analysis
of the data for water quality interpretation. A list of regional and national
references for fish identification is located in Section 8, Fish Bioassessment
Protocols for Use in Streams and Rivers; Section 12, Fisheries Bibliography.
Assistance in confirming questionable identification is available from State,
Federal, and university fishery biologists or ichthyologists. In the Quality
Assurance Project Plan (see Section 2, Quality Assurance and Quality Control),
key(s) used for fish identification should be specified.
Collection No.
Project
Location
Date
Time
Mile
Sampling Device
Collected by
Observations
Preservation(s)
Figure 1. Example of fish sample label information for preserved specimen
container.
6.4 Species Composition (Richness)
6.4.1 A list of species can be compiled using any sampling device, technique,
or combinations of the two. The method used should not select against one or
more species. Also, sampling effort should be thorough enough so that all
species are collected from the study area, and the sampling should be
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conducted several times during the year to include seasonal species. The
calculations for percent species composition in a sample is:
Number of individuals of a given species
„ _ x 100.
Total number of all fish collected
6.5 Length and Weight
6.5.1 Rate of change in length of fish, length frequency distribution, and
weight of fish are important attributes of fish populations. These
measurements can provide an estimation in growth, standing crop, and
production of fish in surface waters.
6.5.1.1 Three length measurements as described by Lagler (1978) are sometimes
used in monitoring studies, but total length is used most often. The three
length measurements (Figure 2) are standard length, fork length, and total
length. Standard length of fish is measured from its most anterior extremity
(mouth closed) to the hidden base of the caudal fin rays, where a groove forms
naturally when the tail is bent from side to side. Fork length is measured
from the most anterior extremity of the fish to the notch in the center of the
tail. It is the center of the fin when the tail is not forked. Total length
is the greatest length of the fish from the anterior most (mouth closed) and
caudal rays squeezed together to give the maximum length measurement. For
fish with a forked tail, the two lobes are squeezed together to give a maximum
length. If the lobes are unequal, the longer lobe is used.
6.5.1.2 A fish measuring board is commonly used to measure length. Fish
measuring boards contain a graduated scale and is usually made of wood or
plastic. Lagler (1978) identifies and discusses factors that can cause
possible errors and inconsistency in taking length measurements. When taking
fish measurements, standard procedures should be written so that the
measurements are done the same way if different individuals are involved in
this procedure.
6.5.1.3 Measurement of fish weight is taken with an accurate scale that can
be used in field studies. Lagler (1978) indicated that precision in weight
measurements is not possible because of variation in the amount of stomach
contents and the amount of water engulfed at capture of the fish. The weights
of live and preserved specimens are not comparable because the percentage of
shrinkage is unknown.
6.5.1.4 Additional information on length, weight, and associated structural
indices are discussed in Anderson and Gutreuter (1983).
6.6 Age, Growth, and Condition
6.6.1 Changes in water quality can, at times, be detected by studying the
age, growth, and condition of fishes taken from a body of water. These
studies require extensive knowledge of the life histories of fish and of the
area being studied, experience in aging fish, sufficient time and manpower to
85
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adequately sample and analyze the data, and sufficient age, growth, and
condition historical data for comparison.
Scale Sample Area
Lateral Line
Standard Length
Fork Length
Total Length
Scale Sample Area
Total Length
Figure 2. Fish measurements (using a fish measuring board) and scale sampling
areas. A. spiny-rayed fish. B. soft-rayed fish. Total length
measurement requires compressed tail to give maximum elongation.
Modified from Lagler (1956).
6.6.2 A problem in using fish for any type of study is their high mobility.
However, Gerking (1959) indicated that many species are relatively sedentary
in summer. Depending on the species, there may be no practical way to
determine with a first time visit how long an individual fish has been in a
given area. Any changes detected in age, growth, or condition are not
necessarily attributable to conditions prevailing at the capture site. Some
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information on fish movement may be obtained from previous State or Federal
studies. Only a carefully planned, long-term study may provide beneficial
data, and only if used in conjunction with other biological, physical, and
chemical data, e.g,, benthic invertebrates (macroinvertebrates), periphyton,
water flow, habitat, and water chemistry.
6.6.3 The methods most commonly used in studying the age and growth of fishes
are: (1) length-frequency, (2) annulus formations in hard parts, such as
otolith, bone, spine rays, and scales.
6.6.3.1 The knowledge of the age and rate of growth of fish is extremely
useful in fishery management. The processes of determining fish age and
assessing fish growth rates are different, but they are closely related and
are usually done at the same time. Table 1 was compiled by the Institute for
Fisheries Research, the University of Michigan, Ann Arbor, Michigan from
samples taken of Michigan fish during a period of approximately 30 years. The
samples were collected mostly during the summer months but all months of the
year are represented. Variations occur among states in sample size according
to species and age groups, and some averages are more reliable than others.
Busacker et al. (1990) discuss various techniques that are used in the study
of fish growth, and they provide guidance to the appropriate uses of specific
growth methods.
6.7 Length-Frequency Method
6.7.1 The length-frequency method for making age determinations is based on
the assumption that fish increase in size with age. When the number of fish
per length is plotted on graph paper for a given species if comparing a
population. Peaks generally appear for each age group.
6.7.2 For this method to provide meaningful data it is important that the
following criteria be met during sampling: (1) the fish must be collected
over a short period; (2) large numbers must be obtained, including fish of all
sizes; (3) the affected area and a control (unaffected) area must be sampled
simultaneously within the same time frame.
6.7.3 For some studies, the length-frequency method may be of limited value
because: (1) it is considered not reliable in aging fish beyond their second
or third growing season (2) acquiring a large number of fish generally
requires several experienced field biologists utilizing different sampling
techniques.
6.8 Length-Age Conversion Method
6.8.1 In certain studies, it may be desirable to know the age of fish of a
given length (e.g., selection data are normally in terms of length, but for
incorporation in yield equations need to be expressed in terms of age.)
Length can be converted to age (Gulland, 1983) by fitting all the observed
data of mean length at age to a growth equation, such as the von Bertalanffy
equation.
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6.8.2 To calculate age (t) in terms of length (1), divide both sides by LB,
and subtract from unity, resulting in
L.
= e
taking natural logs of both sides gives
= -K(t-tQ)
therefore,
where:
t = age (present)
1 = length of individual specimens (length at time (t))
la = maximum length expected for a particular species
t0 = the age at which the fish would be zero size
r = growth rate constant
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TABLE 1. AVERAGE TOTAL LENGTHS IN INCHES FOR EACH AGE GROUP OF SEVERAL FISHES IN MICHIGAN1
CO
Species
Bluegill
Pumpkinseed
Black Crappie
Rock bass
Warmouth
Green sunfish
Largemouth bass
Smallmouth bass
Yellow perch
Walleye
Northern pike
Musekllunge
Smelt
Brook trout
Rainbow trout
(inland lakes and
streams)
Steelhead
(lake-run
rainbow)
0 I
2.3 3.4
. 2.8 3.3
3,6 5.1
1.5 3.1
3.1
3.0
3.6 6.1
3.4 6.1
3.1 4.6
7.1 9,5
10.2 15.6
6.8 15.7
5.3
3.0 6.4
2.2 6.3
13.4
II
4.4
4.4
6.8
4.5
4.4
3.9
8.6
9.2
6.1
13.3
19.4
19.9
6.9
9.0
8.4
17.0
III
5.5
5.2
8.2
5.6
5.2
4.7
10.6
11.3
7.0
15.2
22.2
25.4
7.7
11.5
10.3
18.7
IV
6.4
5.9
9.0
6.5
5.5
5.1
12.2
13.3
8.0
17.2
24.6
31.9
8.1
15.1
11.0
23.6
V
7.0
6.4
9.5
7.4
6.2
5.7
13.6
14.9
9.0
18.6
26.5
34.7
8.8
18.8
25.4
Age Group
VI VII VIII IX X XI XII XIII
7.5 7.9 8.6 8.8 9.1 9.8 9.7
7.9 7.3 7.8 7.4 8.1 9.8
10.6 10.9 11.8 12.2 ...
8.2 8.9 9.6 9.9 10.1 11.6 11.7
6.7 6.9 6.6 7.5 7.3
5.7 5.0
15.1 16.7 17.7 18.8 19.8 19.6 20.8
15.7 16.8 17.5 18.5 19.2 ... 19.2
9.9 10.7 11.3 11.8 12.3 12.3 13.9 13.2
19.2 19.6 21.6 21.4 25.2 23.7 26.5
28.9 32.7 33.4 38.7 39.6 42.0 48.0
36.8 39.2 41.7 45.3 48,7 47.5 49.7
9.6
21.3 23.9 ... ...
28.1 30.0 30.4
1From Laarman (1964), Length of common Michigan sport fishes at successive ages, Michigan Fisheries No. 7,
Department of Fisheries, School of Natural Resources, The University of Michigan, Ann Arbor, MI.
-------
6.9 Annul us Formation Nethod
6.9.1 This technique is based on the fact that fish are poikilothermic
animals and the rate at which their body processes function are affected by
the temperature of the water in which they live. Growth is rapid during the
warm season and slows greatly or stops in winter. This seasonal change
produces a band (annulus) in such hard bony structures as scales, otoliths
(ear stones), fin rays and spines, and vertebrae each year the fish lives.
Scales (Figure 2) are most commonly used in determining the age and yearly
rate of growth because they lengthen throughout the life of the fish at a
predictable ratio to the annual increment in body length. The location of the
body from where the scales are obtained is important. Each species of fish
has a specific body area from which scales should be removed for optimum
clarity and ease of identifying the annuli and a size at which scale formation
begins (Jearld, 1983; Lagler, 1956; Weatherley, 1972). Coin envelopes are
frequently used for holding scales and for recording field data (Figure 3).
Collection No.
Species
Location
Date
Time
Mile
Sampling Device
Collected by
S.I. T.L.
Sex Maturity/and state of organs
Wt.
Annuli
Condition
Figure 3. Example of recording field data information of scale samples for
age and growth studies.
6.9.2 Aging can be accomplished by use of a side-field, low-powered
microscope, but a microprojector is preferred for determining the rate of
growth. Computer assisted microprojectors have been developed for reading
scales more rapidly and accurately.
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6.9.3 It is important that the investigator realize that not all annuli-1 ike
markings are valid. "Spawning-checks", "false annuli", or other annuli-1ike
marks may be present because of disease, body injury, spawning, etc.
6.9.4 The duration of sampling and the number of fish that must be collected
are not as critical as the length-frequency method. Sampling can cover a
considerable period and only a single method need be used for capturing the
fish. Specialized equipment and trained personnel are needed, however, to
identify, analyze, and interpret the data.
6.9.5 To determine any changes in the growth rate of a fish population, it is
essential to use both the length-frequency and annul us methods and have
samples from unaffected localities and/or sufficient background data from the
sampling area. Any changes detected may be attributed to a single or a
combination of natural or man-associated activities that altered the
environment. Some of the most obvious natural modifications are a change in
the average annual water temperature, fluctuating water levels, and
availability of food. Man may also influence the water temperature and
levels, physically alter the environment and fish habitat by damming or
dredging activities, surface mining activities, and introducing substances
that directly or indirectly affect the well-being of the fish population. It
is evident, therefore, that it may be impossible to pin-point what or who was
responsible for the change in the growth rate of a fish population except in a
small lake.
6.10 Condition Factor (Coefficient of Condition)
6.10.1 The condition of fish can be estimated mathematically or by evaluating
physical appearance.
6.10.2 Mathematically, the coefficient of condition is utilized to express
the relative degree of well-being, robustness, plumpness or fatness of fish.
It is based on a length-weight relationship and is calculated by the formula:
Coefficient of Condition KTL = W 105
L3
W = weight in grams
L = length in millimeters
105 = factor to bring the value of K near unity
TL = designation of measuring system used (fork, standard, or
total length)
6.10.2.1 The coefficient of condition is "K" when the metric system is used
in expressing the length and weight, and "C" when the English system is used.
6.10.3 The coefficient of condition has been used by ichthyologists and
fishery biologists to determine the suitability of the environment for a
species. However, it is not recommended for use in short term water quality
studies because any non-environmental factors influence the values derived,
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e.g., changes due to age, sexual differences, and changes with seasons. These
natural fluctuations make it extremely difficult to attribute any change to
the quality of the water from which the fish are collected and must be taken
into account when designing long term studies and evaluating data.
6.10.4 The observance of the physical appearance or condition of fish will
usually indicate the general state of their well being and give some broad
indication of the quality of their environment. When fish are captured they
should be examined to see if they appear emaciated, are diseased, or contain
parasites. The condition of their gills should also be checked. Healthy fish
will be active when handled and are reasonably plump. Dissect a few specimens
and check the internal organs for disease or parasites. The stomach of fish
should also be examined to determine if the fish were actively feeding prior
to capture.
6.10.5 For more detailed information on age, growth, and conditions of fish,
see Anderson and Gutreuter (1983), Bagenal and Tesch (1978), Calhoun (1966),
Carlander (1969), Everhart et al. (1975), Goede (1991), Jearld (1983), Lagler
(1956), Lux (1971), Norman (1951), Ricker (1975), Schram et al. (1992),
Summerfelt (1987), and Weatherley (1972).
6.11 Relative Height Index
6.11.1 Usefulness of typical fisheries metrics for evaluating sensitive
indicator organisms at the population level provide useful information in
comparing subtle differences between sites. The drawbacks to using standard
fisheries approaches are the limitations of either state developed or regional
expectations and the lack of resolution linked with causes. The assessments
require a large sample for site comparison and a large number of reference
stations for determining the expected population regression line. The
traditional approach to the assessment of condition involves the use of a
Fulton-type (Anderson and Guetreuter, 1983) condition factor. This is
calculated as:
K = W/L3
where W is weight (g) and L is length (mm). These factors are both length and
species dependent. Therefore, it is improper to compare fish of different
species or fish of the same species at different lengths. Le Cren (1951)
developed the relative condition factor:
K = W/W x 100
where W is the observed weight and W is the length specific expected weight
for fish in the populations under study as predicted by a weight-length
regression equation calculated for that population. This approach solved the
problem of comparing fish of different lengths and species but, because a
different weight-length regression was calculated for each population,
interpopulational comparisons were not possible. The relative weight (Wr)
index (Wege and Andrson, 1978) enabled interpopulational comparisons by making
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the standard weight-length (Ws) regression species-specific rather than
population specific or location specific. Relative weight is calculated as:
Wr = W/WS x 100
where W is the length-specific standard weight predicted by a weight-length
regression constructed to represent the species as a whole.
6.11.2 Ws equations have defined in most cases to represent populations
in better than average conditions (reference conditions) based on the
assumption that attempting to produce fish populations that attain only
average condition generally does not represent a typical management goal. Ws
should be considered a benchmark for comparison of samples and populations.
Comparisons are based on the 75th percentile of the weight. An alternative
technique, regression-!ine-percent!le (RIP), is based on comparison of
Iog.0weight-log10length regression equations for each population whereas the
typical Wr equation is based on pooled length-weight data.
6.11.3 Murphy et al. (1991) discussed the development of the index and
expounded upon the status and Wr regression equation for 27 species. To
calculate Wr properly requires data from representative or reference stations
over a broad range for the species of interest. Slopes of less than 3.0 are
considered inappropriate for most species because such a slope indicates the
species becomes thinner with increased length. Low slopes may also results
from including small fish in the regression. Differences of weighing small
fishes and the inherent problems of weighing small fishes in the field may
preclude development of a single equation for an entire species life history.
A minimum applicable length is to determine the minimum size which should
be weighed. For other species the minimum length is a function of the
variance:mean ration for Iog10 weight where it sharply increased.
6.13 Literature Ci
Anderson, R. and S.J. Gutreuter, 1983, Length, weight, and associated
structural indices. In: Nielsen, L.A. and D.L. Johnson (eds.).
Fisheries Technique. Amer. Fish. Soc., Bethesda, MD. pp. 283-300.
Bagenal, T. B, 1978, for assessment of fish production in fresh
waters. IBP Handbook No, 3, Blackwell Sci. Pub!., Oxford, England.
Bagenal, T.B. and F.W. Tesch. 1978. Age and growth. Pages 101-136. In:
Bagenal, T.B. (ed.). for assessment of fish production in
fresh waters. IBP Handbook No. 3. Blackwell Sci. Publ., Oxford,
England.
Busacker, G.P., I.R. and E.M.Goolish, 1990. Growth. In: C.B.
Schreck and P.B. Moyle {eds.}. Methods for fish biology. Amer. Fish.
Soc.» MD.' pp. 363-387.
Calhoun, A. (ed.). 1966. Inland fisheries management. Calif. Dept. fish and
Game, Sacramento, CA.
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Carlander, K.D. 1969. Handbook of freshwater fishery biology. Vol. 1. Iowa
state Univ. Press, Ames, IA.
Everhart, W.H., A.M. Eipper, and W.D. Young. 1975. Principles of fishery
science. Cornell Univ. Press, Ithaca, NY.
Gerking, S.D. 1959. The restricted movement of fish populations. Biol.
Review 34:221-142.
Goede, R.W. 1991. Fish health/condition assessment procedures. Utah
Division wildlife Resources, Fisheries Experiment Station, 1465 West 200
North, Logan, UT. 29 pages.
Gulland, J.A. 1983. Fish stock assessment: a manual of basic methods.
FAO/Wiley Series, Vol. 1. Wiley & Sons, NY. 223 pp.
Jearld, A., Jr. 1983. Age determination. In: Nielsen, L.A. and D.L.
Johnson (eds.). Fisheries Technique. American Fisheries Society,
Bethesda, MD. pp. 301-324.
Lagler, K.F. 1956. Freshwater fishery biology, 2nd. Edition. William C.
Brown Co., Dubuque, IA.
Lagler, K.F. 1978. Capture, sampling and examination of fishes. Pages
7-47. In: T.B. Bagenal (ed.). Methods for assessment of fish
production in fresh waters. IBP Handbook No. 3. Blackwell Sci. Publ.,
Oxford, England.
Laarman, P.M. 1964. Length of common Michigan Sport Fishes at successive
ages. Michigan Fisheries No. 7, Department of Fisheries, School of
Natural Resources, The University of Michigan, Ann Arbor, MI.
LeCren, E.D. 1951. The length-weight relationship and seasonal cycle in
gonad weight and condition in the perch (Perca fluviati7/s). J. Animal
Ecol. 20(2):201-219.
Lux, F. 1971. Age determination in fishes. U.S. Fish & Wildlife Ser.,
Fishery Leaflet No!. 637, Washington, DC.
Matthews, W.J. 1985. Distribution of midwestern fish on multivariate
environmental gradients, with emphasis on Notropis lutrensis. Amer.
Midi. Nat. 113:225-237.
Matthews, W.J. and H.W. Robison. 1988, The distribution of the fishes of
Arkansas: a multivariate analysis. Copeia 1988:358-374.
Mayden, R.W. 2985. Biogeography of Ouachita Highland fishes. Southwestern
Nat. 30:195-211.
Mayden, R.W. 1988. Vicariance biogeography, parsimony, and evolution in
North American fishes. Syst. Zool. 37:329-355.
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McAllister, D.E., S.P. Platania, F.W. Schueler, M.E. Baldwin, and D.S. Lee.
1986. Ichthyofaunal patterns on a geographic grid. In: C.H. Hocutt
and E.O. Wiley (eds.). The zoogeography of North American freshwater
fishes. John Wiley and Sons, Inc., New York, NY.
Murphy, B.R., D.W. Willis, and T.A. Springer 1991. The relative weight
index in fisheries management: status and needs. Fisheries 16(2):30-
38.
Nielsen, L.A. and D.L.Johnson (eds.). 1983. Fisheries Techniques. Amer.
Fish. Soc., Bethesda, MD. 468 pp.
Norman, V.R. 1951. A history of fishes. A.A. Wyn Inc., New York, NY.
Ricker, W.E. 1975. Computation and interpretation of biological
statistics of fish populations. Bull. Fish. Res. Board. Can. 191.
382 pp.
Schram, S.T., T.L. Margenau, and W.H. Blust. 1992. Population biology and
management of the walleye in western Lake Superior. Technical Bulletin
No. 177, Department of Natural Resources, Madison, WI. 28 pp.
Schreck, C.B. and P.B. Moyle (eds.). 1990. Methods for fish biology. Amer.
Fish. Soc., Bethesda, MD.
Summerfelt, R.C. 1987. Age and growth of fish. Iowa State University, Ames,
IA.
USEPA. 1990. Biological criteria. National program guidance for surface
waters. EPA-440/5-90-004. Office of Water Regulations and Standards,
U.S. Environmental Protection Agency, Washington, DC.
USEPA. 1991. Biological Criteria. State Development and Implementation
efforts. EPA-440/5-91-003. Office of Water, U.S. Environmental
Protection Agency, Washington, DC.
Weatherley, A.M. 1972. Growth and ecology of fish populations. Academic
Press, NY, NY.
Wege, G.J. and R.O. Anderson. 1978. Relative weight (Wr): a new index of
condition for largemouth bass. In: G. Novinger and J. Dillard (eds.).
New approaches to the management of small impoundments. Amer. Fish.
Soc., North Central Division, Special Publication 5, Bethesda, MD. pp.
79-91.
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SECTION 7
SPECIAL TECHNIQUES
7.1 Flesh Tainting (Flavor Impairment)
7.1.1. Sublethal concentrations of chemicals, such as phenols, benzene, oil,
and 2,4-D, are often responsible for imparting an unpleasant taste to fish
flesh, even when present in very low concentrations.
7.1.2 Specific methods have been developed by Thomas (1969), APHA (1992), and
ASTM (1992) in which untainted fish are placed in cages or exposure tanks
upstream and downstream or in the laboratory from suspected waste water
sources. The techniques in these references and Subsection 7.1.3 should
successfully relate the unacceptable flavor produced in exposed native fish to
a particular waste source.
7.1.3 The following procedures are presented as a working guide for fish
flesh tainting or flavor impairment.
7.1.3.1 To ensure uniform taste quality before exposure, all fish are held in
pollution-free water for a 10 day period. After this period, a minimum of
three fish are cleaned and frozen with dry ice as control fish. Test fish are
then transferred to the test sites, and a minimum of three fish are placed in
each portable cage. The cages are suspended at a depth of 0.6 meter for 48 to
96 hours.
7.1.3.2 After exposure, the fish are filleted, frozen on dry ice, and stored
at 0°C until tested. The control and exposed samples are shipped to a fish
tasting panel, such as is available at the food science and technology
departments in many major universities, and treated as follows: (1) The fish
are washed, wrapped in aluminum foil, placed on slotted, broiler-type pans,
and cooked in a gas oven at 218°C (400 F) for 23 to 45 minutes depending on
the size of the fish; (2) Each sample is boned and the flesh is flaked and
mixed to ensure a uniform sample; (3) The samples are served in coded cups to
judges. Known and coded references or control samples are included in each
test. The judges score the flavor and desirability of each sample on a point
scale. The tasting agency will establish a point on the scale designated as
the acceptable and desirable level.
7.2 Fish Kill Investigations
7.2.1 Fish kills in natural waters, though unfortunate, can in many instances
indicate poor water quality and environmental health leading to investigations
which may improve the water quality. Prompt investigations should be
organized and conducted so that the resultant data implicates the correct
cause. Fish kills tend to be highly controversial, usually involving the
general public as well as a number of agencies. Therefore, the
investigator(s) can expect his finding to be disputed, quite possibly in a
court of law.
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7.2.2 Possible Fish Kill Sources
7.2.2.1 Fish mortalities result from a variety of causes, including natural
and man-induced. Possible natural fish kills are caused by phenomena such as
acute temperature change, storms, ice and snow cover, decomposition of natural
organic materials, salinity changes, spawning mortalities, and parasitic,
bacterial, and viral epidemics. Man-induced fish kills may be attributed to
municipal or industrial wastes, agricultural activities, and water
manipulations.
7.2.2.2 Winter kills occur in northern areas where ice on shallow lakes and
ponds becomes covered with snow, and the resulting opaqueness stops
photosynthesis. The algae and vascular plants die because of insufficient
light, and plant decomposition results in oxygen depletion. Oxygen depletion
and extreme pH variation can also be caused by the respiration or decay of
algae and higher plants during summer months in very warm weather. Fish kills
resulting from such causes are often associated with a series of cloudy days
that follow a period of hot, dry, sunny days. Fish kills also occur in rivers
below high dams immediately following the opening of a gate permitting cold
hypolimnionic water to flow into the streams as in the Tennessee Valley
Authority (TVA) region.
7.2.2.3 Temperature changes, either natural or the result of a heated water
discharge may result in fish kills. Long periods of very warm, dry weather
may raise water temperatures above lethal levels for sensitive species. A
wind-induced seiche may be hazardous to certain temperature sensitive, deep-
lake, cold-water fish, or fish of shallow coastal waters. Lake water
inversion during vernal or autumnal turnover may result in toxic materials or
oxygen-free water being brought to the surface. Interval seiche movement in
which a toxic or low dissolved oxygen hypolimion flows up into a bay or bayou
for a limited period of time, and later returns to normal levels may also
cause fish kills.
7.2.2.4 Disease, a dense infestation of parasites, infection from bacteria,
or viruses, or natural death of weakened fish at spawning time must always be
suspected as contributory factors in fish mortalities.
7.2.2.5 Occasionally fish may be killed by toxins released from certain
species of living or decaying algae that reached high population densities
because of the increased fertility resulting from organic and inorganic
pollution.
7.2.2.6 Investigations in Tennessee have shown that the leaking of small
amounts of very toxic chemical from spent pesticide-containing barrels used as
floats for piers and diving rafts in lakes and reservoirs can produce
extensive fish kills (TVA, 1968).
7.2.2.7 Industrial waste discharges and waste discharges from a municipal or
domestic type sewerage system may be potential sources of fish kills. These
wastes may be subjected to treatment of a municipal treatment plant or may be
discharged directly, untreated, to a stream. Generally, the municipality or
owner of the sewerage system is held responsible for any discharge in such a
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system; consequently, after collecting samples, the owner or a representative
of the owner of the sewerage system should be contacted. This may be a sewage
treatment plant operator, city engineer, public works supervisor, a
subdivision developer, etc. If the cause of the fish kill was the result of
an industrial waste discharge to a municipal sewer and thence to a stream,
information should be obtained from a municipal official about the industry
and the problem. This should be done only in cooperation with a municipal
official.
7.2.2.8 Pollution capable of causing fish kills may result from such
agricultural operations as pesticide dusting and fertilizer applications, as
well as manure or other organic material discharges to a stream. Generally,
fish kills related to these factors will be associated with rains and runoff.
The source or type of pollution may be difficult or impossible to locate
exactly because it may involve a large area. Talking to local residents may
help pinpoint the specific problem area. Runoff from fields, drainage
ditches, and small streams leading to the kill area are possible sampling
places which may be used to trace the causes.
7.2.2.9 Temporary or intermittent activities, such as mosquito spraying,
construction activities involving chemicals, other toxic substance, and
herbicide containing materials toxic to fish such as arsenic, are also
potential causes of fish kills. As with agricultural activities, tracing the
cause of these kills is difficult and may require extensive sampling.
Accidental spills from ruptured tank cars, pipelines, etc., and dike collapse
of industrial pond dikes are frequently sources of fish kills.
7.2.3 Types and Extent of Fish Kills
7.2.3.1 One dead fish in a stream may be called a fish kill. However, in a
practical sense some minimal number of dead fish observed plus additional
qualifications should be used in reporting and classifying fish kill
investigations (USEPA, 1973). These qualifications are based on a stream
approximating 200 feet in width and 6 feet in depth. For other size streams,
adjustments should be made.
7.2.3.2 Minor fish kills (1-100 dead or dying fish) may be considered "no
fish kill" if confined to a small area or stream reach provided this is not a
recurring event. For example, fish kill occurring near a waste water outfall
in which stream dilution mitigates the effect of the deleterious material. If
this is a recurring situation, it could be of major significance and should be
investigated.
7.2.3.3 Moderate fish kill {100 - 1000 dead or dying fish) may be considered
to have occurred if a number of species and individuals habe been affected in
1-2 km of stream where dilution would have been expected to play a mitigating
role. Apparently normal fish may be collected immediately downstream from the
observed fish kill area.
7.2.3.4 Major fish kill (1000 - 10,000 fish or more dead or dying fish) may
be considered to have occurred in 10-20 km of a stream in which dilution would
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have been expected to have a mitigating effect and when many species of fish
are affected and dying fish may still be observed downstream.
7.2.4 Preparation for Field Investigation
7.2.4.1 All possible speed must be exercised in conducting the initial phases
of any fish kill investigation because fish disintegrate rapidly in hot
weather, and the cause of death may disappear or become unidentifiable within
a short period of time. Success in solving a fish kill problem is usually
related to the speed with which investigators can arrive at the scene after a
fish kill begins. The speed of response in the initial investigation is
enhanced through the training of qualified personnel who will report
immediately the location of observed kills, the time that the kill was first
observed, the general kinds of organisms affected, an estimate of the number
of dead fish involved, and any unusual phenomena associated with the kill.
7.2.4.2 Because there is always the possibility of legal liability associated
with a fish kill, lawyers, judges, and juries may scrutinize the investigation
report. Therefore, the investigation must be made with great care. When
investigating a fish kill, a specific litigation or case number should be
assigned and used on all labels, field data sheets, photographs, and other
records related to the fish kill investigation. Table 1 is a general
flowchart to help with the coordination of a fish kill investigation.
7.2.5 Legal Aspects
7.2.5.1 A chain-of-custody (see Section 2, Quality Assurance and Quality
Control) must be adhered to when any fish kill is investigated and samples
collected for analysis and presentation as evidence. If care is not taken to
establish the validity of samples collected in the field and transported to a
laboratory for analysis, potential evidence for a court action may be lost or
ruled invalid.
7.2.5.2 Several types of evidence including oral and hearsay, circumstantial,
and graphic may be collected during an investigation. Oral and hearsay
evidence should be signed and dated by the individual giving the information.
Circumstantial evidence must be carefully documented as to methods of
collection, who collected it, and disposition of the evidence. Graphic
evidence such as photographs should be accompanied by data listing when taken,
how, by whom, the type of camera and film used, and who processed the film.
7.2.5.3 All samples must be handled in a similar orderly procedure and a
complete record should be kept on their disposition. Recognized tests should
be used and such tests must be approved in detail by USEPA or other recognized
authorities. New test methods must be technically defensible. All unused
portions of samples must be saved until released by the USEPA attorney working
on the case.
7.2.5.4 The investigative team should make every effort to educate the
attorney handling the case. The attorney should be aware of the expertise of
the team, the methods used, validity of evidence collected, and complete
disposition of the evidence.
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TABLE 1. FLOWCHART FOR THE COORDINATION OF A FISH KILL INVESTIGATION1
Pollution Report
Designated investigator (any agency)
Oil or hazardous
material-notify
Completes pollution
report form—notify
USEPA or USCG
Oil or hazardous
material
notify
State Water
Quality Agency
(Conventional pollutants and ^^"^ Fish killed-
in absence of , USEPA or USCG > notify
On-site coordinator
On-site coordinator
State Fish and Wildlife
Resources Agency _
First on-site investigator
Oil spills
Requisition and place
materials for initial
mitigation.
Complete
mitigation
Area reconnaissance to determine:
1. Possible mitigation strategies
to limit pollution impact
2. Beginning and apparent end points
of pollution
3. Relative magnitude of the problem
Conventional
pollutants or
natural causes
Collect preliminary chemical,
physical, and biological
samples in affected and
unaffected areas to document
nature and source of pollutant.
Hazardous materials
CAUTION! Await arrival
of designated eaergency
response personnel.
USEPA
or USCG
State Water Quality
Agency
Field and laboratory measurements
to determine and document nature
and source of the pollutant
State Fish and Wildlife
Agency
Fish kil count for
resource damage and
monetary value assessment
Regulatory action
Modified from Meyer and Barclay (1990). Abbreviations: U.S.
Environmental Protection Agency (USEPA), U.S. Coast Guard (USCG)
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7.2.6 Field Investigations
7.2.6.1 The following is a brief discussion of a suggested method of field
investigation (Meyer and Barclay (1990) provide guidelines, detailed, and
specific procedures for fish kill field investigation). For additional
methods, see the following references: AMPA (1992), Amer. Fish. Soc. (1982),
ASTM (1992), Burdick (1965), Hill (1983), Smith et al. (1956), Tracy and
Bernhardt (1972), U.S. Dept. Interior (1970), USEPA (1973), USEPA (1979a,b),
USEPA (1980), and Section 12, Fisheries Bibliography, Subsection 12.7 Fish
Kills.
7.2.6.2 Individuals involved in fish kill investigations should have a copy
or be familiar with the document, Field Manual for the Investigation of Fish
Kills, (F.P. Meyer and L.A. Barclay, eds., 1990). This document contains
detailed information on the following: planning the investigation (Hunn,
1990), interpreting the fish kill location (Meyer and Herman, 1990), toxic
substances effects and diagnosis (Hunn and Schnick, 1990), fish kills due to
natural causes (Herman and Meyer, 1990), role of infectious agents in fish
kills (Herman, 1990), quality assurance and legal requirements (Schnick,
1990a), where to send samples for analyses (Schnick, 1990b), shipping samples
(Barclay, 1990a), writing the fish kill report (Meyer, 1990a), preparing for
legal testimony (Barclay, 1990b), specific equipment needed for field
assessments (Ardinger, 1990), and case histories of fish kills (Meyer, 1990b).
7.2.6.3 Since the speed with which an investigative team arrives at a fish
kill is extremely important, a few advanced preparations are necessary. The
public should be aware of whom to contact and where to report fish kills. If
possible, a Region- or State-wide network of designated fish kill
investigators should be established, each representing an area in which an
investigator knows the water, biota, and potential polluters. In preparation
for quick action, an investigator must have at his/her immediate disposal:
telephone report sheets (Table 2), a checklist of equipment items (Table 3),
maps of the area, and a list of cooperating analytical laboratories.
7.2.6.4 Make a reconnaissance of the kill area. Make a decision as to the
extent of the kill and if a legitimate kill really has occurred. If a
legitimate kill exists take steps to trace or determine the cause. Secure
sampling equipment and determine size of investigative team needed. Standard
equipment should be taken on all investigations (Table 3), and a standard
checklist with space for special equipment will often save embarrassment in
the field. The on-site study includes specific field observations (Table 4)
that may be made on a fish kill form (Table 5). In addition, specific field
observations (Table 4) should be emphasized, and complete weather data should
be collected (for the period) prior to and during the fish kill. Water
conditions both in and outside the affected area should be noted (i.e.,
appearance of water, turbidity, algal blooms, oil, unusual appearance, etc.).
Stream flow patterns (i.e., high or low flow, stagnant or rapidly moving
water, tide moving in or out, etc.) should be noted and recorded. If
possible, obtain discharge reading from stream gauge if one is near fish kill
area. During the initial steps of the investigation, water chemistry and
physical parameters (e.g., pH, dissolved oxygen, temperature, specific
conductance, and flow) must be determined immediately upon arrival at the kill
101
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site. While none of these factors may be directly involved in the fish kill,
these tests are simply and rapidly performed in the field and can be used as a
baseline for isolating the cause(s) of the kill. Make a rough sketch or
define the fish kill area on a map so that sampling points, sewer outfalls,
etc. can be accurately located on a drawing to be included in a final report.
Take close-up and distance photographs of the dead fish in the stream in the
polluted area, the stream above the polluted area, and the wastewater
discharges. Photographs will often show a marked delineation between the
wastewater discharge and the natural flow of water. Pictures taken at a
relatively high elevation, (a bridge as opposed to a boat or from a low river
bank) will show more and be more effective. Color photographs are also more
effective than black and white prints in showing physical conditions of a
stream.
7.2.6.5 Certain biological observations should also be made as soon as
possible: (1) the presence or absence of plankton blooms, (2) dead or living
macroinvertebrates and fish, and (3) the actions of moribund fish. Additional
observations are listed in Tables 6, 7, and 8.
7.2.6.6 The location of sampling stations is very important. If there are no
obvious reasons for a kill, stations should be selected in and outside the
apparent kill area. If there are possible polluters, each should be suspect
and sampling stations must be selected within and outside of the area of
influence for each possible suspect.
7.2.6.7 In flowing waters, where a pollutant may be discharged as a slug, the
investigator should try to estimate the time of kill, determine stream
velocity, and collect samples downstream in the vicinity of the slug.
7.2.6.8 Water samples must be collected and processed in a variety of ways
depending on the types of analyses required. An updated USEPA methods list
for collection and preservation of samples should be at the disposal of the
investigator (USEPA, 1979a,b).
7.2.6.9 The collection and preservation of aquatic organisms may require
special techniques. For example, it is always best, if possible, to collect
moribund fish from the affected area. If none are available, freshly dead
fish will have to be utilized. Unaffected fish from outside the kill area
must also be collected. All samples should be handled with regard to the type
of suspected toxicant and the type of analysis to be performed.
7.2.6.10 Contact personnel from the laboratory or laboratories which will
participate in analyzing samples. If possible estimate the following and
record on the fish kill general information form (Table 2).
1. The number and size of samples to be submitted.
2. The probable number and types of analyses required.
3. The dates the samples will be received by the laboratory.
4. Method of shipment to the laboratory.
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TABLE 2. FISH KILL GENERAL INFORMATION FORM
1. Who is the informant?
Name Phone
Address
Directions to meeting place
Date and Time
2. Reporting Source
Agency
Address
Phone(s)
Fish Kill Network Yes NO
3. Location of kill (county, town, access point):
4. Duration of kill: First noticed - Time Date
Is it continuing? (Yes, No). If not, when did it stop _
5. Extent of Kill: Area covered (miles of stream or size of pond or lake)
6. Approximate number of fish affected Species
Size (length, age classes)
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TABLE 2. FISH KILL GENERAL INFORMATION FORM (CONTINUED)
7. Opinion as to cause
8, Recent activities (crop dusting, weather change, etc.)
9. Possible sources of pollution
10. Measures taken
11. Action Requested
Field Investigation
Laboratory Analyses
12. Assistance to Project
Provided by
Personnel
Equipment
Transportation Facilities
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TABLE 3. CHECKLIST OF FISH KILL INVESTIGATION EQUIPMENT
General
Fish
1 . Boat
2. Motor
3. Paddles
4. Life preservers
5. Camera
6. Film
7. Ice chests
8. Wet ice
9. Dry ice
10. Portable light source
11. Waterproof notebook
12. Waterproof pencils
13. Waterproof labels
14. Chain of custody seals
15. Chain of custody forms
16. Arm-length gloves
17. Hip boots
18. Chest waders
19. Paper towels
20. Aluminum foil
21. Thermometer
22. Plastic bags,
23. DO kit
24. pH equipment (probe,
25. Glass jars (chemical
26. Maps
27. Hand tallys
28. Tape measure (100 yd
29. Rain gear
30. Polaroid glasses
31. Tamper proof seals
assorted sizes
colorimeter)
samples)
or meter)
1. Dipnets
2. Seines
3. Nets
4. Electrofishing gear
(if available)
5. Weight scale
6. Measuring board
7. Tubs
8. Fish counting forms
9. Dissecting kit
10. Heparinized vials
11. 10% formalin
12. 70-75% ethyl alcohol (ethanol)
13. Scale envelopes
Benthos
1. Ekman grab sampler
2. Ponar grab sampler
3. Surber-type sampler
4. Drift net sampler
5. Dipnets, kick nets
6. Quart and pint widemouth
containers
7. 70-75% alcohol
8. 10% formalin
9. Foot tub
10. U.S. Standard 30 sieve
11. Forceps
PIankton-Peri phyhton
1. Water sampler - Van Dorn
2. Vials, small widemouth jars
3. 6-3-1, Formalin preservative
4. 2-liter jars
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TABLE 4, FIELD OBSERVATIONS
1. Locate the kill area.
2. Take water samples for chemical analysis and preserve properly.
3. Make chemical and physical field analyses (DO, pH, temperature, flow,
weather, etc.).
4. Record observations (odors, flocks, sheens, deposits, etc.).
5. Collect fish for analyses (follow guidelines for various analyses).
6. Collect plankton samples.
7. Collect periphyton.
8. Collect macroinvertebrates (substrate, drifting, and attached).
9. Extensive and pertinent observations:
* Observe and examine dead and dying fish (see other attachment.
* Are small fish collected in tributaries--on surface or not?
* Are the plankters (planktonic organisms) concentrated in the kill
area?
* Are they alive and viable or dead?
* Is there extensive periphytic growth?
* Is the benthic community active, over-active, quiescent?
* Are there many drifting organisms?
* Record all observations.
10. Repeat the applicable steps above in a non-affected area of the lake or
stream.
11. Take numerous pictures of the overall area, specific problem areas, dying
fish, algae blooms, water conditions (color, turbidity, etc.).
12. Counts of mortality by species to estimate resource loss.
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TABLE 5. FISH KILL INVESTIGATION FORK
Stream, Lake, Other
Stream Mile
County
Drainage Basin
Tributary to
State
Top. Map.
Nearest Town
Highways
Fish Kill Began: Time
Date
Time and Date of First Report
Address
Investigators: (Name and Agency)
Ended: Time
Reported by (Name)
_ Telephone
Date
Area Affected: Upstream Limits
Downstream
Miles
Weather Conditions: Present
Past 48 Hours
Photographic Record:
Field Measurements:
Acres
Picture No.
Time and Date
Sub.iect
Sample
Temp.
DH
DO
Conductivity
Gaqe Ht./Flow
Uostream of Kill
In Kill Area
Downstream of Kill
Comments and Possible Sources:
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TABLE 5. FISH KILL INVESTIGATION FORM (CONTINUED)
Location of Collected Samples
Sample ID. No.
Station Description
Sample ID No:
Sample
Water Sample*
Fish (frozen)
Fish (formalin)
Fish (fresh)
Fish Blood:
Species lenqth
Species lenqth
Species lenqth
Species lenqth
Sediment
Alqae (frozen)
Alqae (iced)
Benthos
Special Analysis
Upstream of Kill
In Kill Area
Downstream of Kill
Approximate I of dead fish of (of each species)/acre, mile, 100 yards, etc.
Requested Analyses:
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TABLE 6, OBSERVATION ON DEAD AND MORIBUND FISH
External
1, External examination for fungus, bacteria, open sores, parasites.
2. Gill examination for color, abnormal morphology, gill lice, slime,
collapse of filaments, adhesion of filaments,
3. Eyes opaque, clear, covered by mucus.
4. Fins - anchor lice, extended-folded, bleeding, fungused, frayed.
5. Scales - loose groups, bent, bleeding, missing.
6. Body - bent, twisted, rigid.
7. Mouth - open, normal, hyper-extended in death.
Internal
1. Do they bleed freely?
2. Is the liver clear of spots or open lesions? Is it a light off-brown
or tan?
3, Is the air bladder hard, very soft, or partly inflated?
4. Is the stomach full or empty? What is in it?
5. Is the entire intestinal tract empty?
6. Are there internal parasites in the abdominal cavity?
7, Is there watery fluid in the abdomen?
8. Is there discoloration of any of the tissues?
9, Are the muscles pulled away from the ribs or backbone?
10. Are there lesions or spots in the muscles? Describe them.
11. Is the kidney (against the backbone) a normal dark red to purple or
unspotted?
12. Are there lesions or watery abscesses (i.e., blisters)?
13. Is the pericardia! space free with watery fluid or is it discolored a
reddish or yellow color?
14. Are the fish slimy or dry?
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TABLE 6. OBSERVATION ON DEAD AND MORIBUND FISH (CONTINUED)
Internal (continued)
15. Are there trailing mucus strings from the gills or fins?
16. Are there large patches of missing scales?
17. Is there bleeding about the fin bases or scale bases?
18. Do the gills look very bright red, dark blue, or purple? Are the gills
covered with slime? Are they bleeding or lumpy?
19. Do the gill covers move very rapidly or very slowly?
20. Are the fish unresponsive, roll over in the water, and slowly die? Do
they slowly settle to the bottom while upright?
21. Do any rest upside down at the surface and still breathe?
22. Do any cough, flare the gill covers, or flare the fins?
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TABLE 7. OBSERVATIONS ON EFFECTED FISH
1. Do the fish swim wildly at the surface? If they do, do they do it
continuously or in erratic and irregular bursts of activity?
2. Do they try to leap from the water after racing across the surface?
3. After they race at the surface, do they fall on their side and tremble?
4, How long do they race about?
5. Do they race about, then tremble, turn over, and die , or do they race,
rest, then race with increasing periods between bursts of activity?
6. At the end of a run, are the bodies twisted or rigidly bent to one side
or the other?
7. As activity decreases, do they rest upright at the surface?
8. As activity decreases, do they rest head-down in the water?
9. As activity decreases, do they rest tail-down in the water?
10. As activity decreases, do they rest tail-down in the water and spin on
their long axis?
11. With the slower erratic swimming, do they swim forward, slowly turning
over and over, spiral ing, or swim forward but describe a long curving arc
or circle?
12, Do they swim slowly forward, mouthing at the surface with audible
"smacking" sounds?
13. Do they swim slowly forward, ejecting bubbles from the mouth?
14. As swimming slows or ceases, do they settle into the water or do they
struggle to stay down and upright?
15. If you can catch them, must you use a net, or can you catch them by hand?
16. Once caught, do they struggle, tremble, lose scales, or go rigid?
17. Are they bleached out, very dark, or blotchy?
18. Are there fuzzy blotches anywhere on the body?
19. Are there open scores?
20. Are the fins and gill covers folded or held rapidly extended from the
body?
Ill
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TABLE 7. OBSERVATIONS ON EFFECTED FISH (CONTINUED)
21. Are they slimy or dry?
22, Are there trailing mucus strings from the gills and fins?
23. Are there large patches of missing scales?
24, Is there bleeding about the fin bases or scale bases?
25. Do the gills look very bright red, dark blue, or purple? Are the gills
covered with slime? Are they bleeding or lumpy?
26. Do the gill covers move very rapidly or very slowly?
27. Are the fish unresponsive, roll over in the water, and slowly die?
28. Do they slowly settle to the bottom while upright.
29. Do any rest upside down at the surface and still breathe?
30. Do any cough, flare the gill covers, or flare the fins?
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TABLE 8. SYMPTOMS THAT HAVE BEEN RELATED TO CAUSE OF FISH DEATH1
SYMPTOM
CAUSATIVE AGENTS
Gasping at surface
Fish dying in early morning only
Swimming slowly in circles
or only one species affected
Erratic swimming patterns,
contorted bodies, tremors,
or convulsions. Other animals
involved (i.e., birds,snakes,
turtles, etc.)
Fish gills covered with mucus,
or clogged
Small fish kills of various
species over a long period
of time, altered species
composition
Deflated swim bladders and
viscera obliterated
White film on gills, skin
and mouth
Sloughing of gill epithelium
Gill occlusion
Bright red gills
Dark gills
Gill lamellae thickening
Distended gill covers
Low DO or rotenone
Low DO, summer kill
Disease
Pesticides
Rotenone, high suspended
solids, heavy metals
Low concentrations of trace
metals
Seismic blasts, dynamite, or
other explosives.
Acids, heavy metals.
trinitrophenol
Copper, zinc, lead, detergent,
ammonia, quinoline
Turbidity, ferric hydroxide
precipitate
Cyanide
Phenolic poisoning, p-cresol,
naphthalene, oxygen deficiency
Hydrogen sulfide
Ammonia, cyanide
Modified from Janet Kuelfer, USEPA, Region 9, San Francisco, CA.
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TABLE 8. SYMPTOMS THAT HAVE BEEN RELATED TO CAUSE OF FISH DEATH (CONTINUED)
SYMPTOM
CAUSATIVE AGENTS
Swollen abdomens
Blue stomachs
Intestinal epithelium
destruction
Gall bladder distension
Extreme thinning of
stomach wall
Pin point white spots,
fish rubbing against substrate
Chlorinated hydrocarbon,
insecticides
Molybdenum
Hexavalent chromium,
pulp mill wastes
Pulp mill wastes
Endosulfan
Ichthyophonus sp., Cryptocaryon sp,
(Ich disease)
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5. To whom the laboratory results are to be reported.
6. The date the results are needed.
7.2.7 General Sampling Procedures (also see Meyer and Barclay, 1990). The
extent and method of sampling will depend upon location and upon the suspected
cause of the kill.
7.2J.I For stream and wastewater sampling, sample the following points when
the pollution discharge is coming from a well defined outfall:
1. The effluent discharge outfall.
2. The stream at the closet point above the outfall which is not influenced
by the waste discharge.
3. The stream, immediately below the outfall.
4. Other points downstream needed to trace the extent of the pollution.
7.2.7.2 The sampling should be extensive enough that when all the data is
compiled no question will exist as to the source of the pollution which killed
the fish.
1. Streams less than 200 feet wide, not in an industrial area usually can be
adequately sampled at one point in a section (Figure 1).
2. Streams 200 feet or wider generally should be sampled two or more places
in a section immediately above and below the pollution discharge. Where
the pollutional waste has adequately mixed with the stream flow one sample
may suffice.
3. A number of samples in a cross section may be required on any size of
stream to show that the suspected pollutional discharge is coming from a
source located in an industrial or municipal complex (Figure 2).
4. Extensive cross sectional sampling on rivers greater than 2000 feet wide
will be required for kills involving suspected agricultural or other types
of mass runoff.
5. Sample depth - on streams 5 feet in depth or less, one mid-depth sample
per sampling location is sufficient. For streams of greater depths,
appropriate sampling judgment should be used since stratification may be
present.
7.2.7.3 The number of samples to be collected at a given cross section will
depend principally on the size of the stream.
a. Ten 1 L water samples should be collected from the kill area for
chemical analyses as well as other 1 L samples from control and
other stations. (In flowing waters samples should also be collected
in the estimated location of the main slug).
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b. Ten pounds including ten individuals of dying fish of each important
species frozen with dry ice. An equivalent amount and number of
control fish.
c. Five small fish of each important species preserved in formalin.
d. Five dying fish of each significant species placed on wet ice and
delivered to a fish disease laboratory within 24 hours. (Some fish
disease labs specify fish placed in bags next to wet ice.)
e. A minimum of ten fish should be collected for histochemical
analysis. Refer to Section 5, Fish Specimen Processing, on the
proper fixation and preservation of fish tissues for histochemistry
methods.
f. Five vials containing 5 cc. each of blood from each important
species.
g. Ten gallons of water for bioassay.
h. One quart to one gallon of sludge or sediment.
i. Ten cc. of concentrated algae frozen.
j. Ten cc. of concentrated algae chilled.
k. Benthic invertebrate (macroinvertebrates) samples.
7.2.8 Explanation of Figures 1 and 2.
7.2.8.1 Collection point 1 (Figure 1) and points 3 and 4 (Figure 2) should be
collected as near to the point of pollutional discharge as possible. These
points will vary according to stream flow conditions. The pollution
discharges into a slow sluggish stream usually will have a cone of influence
upstream of the outfall; whereas, a swift flowing stream usually will not.
7.2.8.2 Collecting an upstream control sample from a bridge within sight of
the pollutional discharge would probably be satisfactory in Figure 1 but
definitely not in Figure 2.
7,2.8.3 Figures 1 and 2 are given for illustrative purposes only and should
be used only as a guide for sampling. Each individual situation must be
individually considered to insure adequate, proper sampling. While too many
samples are better than too few, effort should be made not to unduly overload
the laboratory with samples collected as a result of poor sampling procedures.
7.2.9 Biological Sampling
7.2.9.1 In every investigation of fish kills the paramount item should be the
immediate collection of the dying or only recently dead organisms. Sampling
and preservation are as follows:
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Suspected source of
pollution..
)
Direction
flow.
fa
Of 2 ^
8 M
OJ °7
• Area of dead fish and/or
obvious pollution discharge.
1 L.
Bridge •
Figure 1, Minimum water sampling point on stream 200 feet or less wide
involving an isolated discharge. Modified from USEPA (1973).
Dlscnorqa sources relatively ciose
to suspected source of pollution. \_
Direction of
flow
Suspected source
of pollution.
Figure 2. Minimum water sampling points on a stream running through an
industrial or municipal complex. Modified from USEPA (1973).
117
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1. Collect 20+ drops of blood in a solvent rinsed vial, seal with aluminum
foil, cap, and freeze.
2. Place bleeding specimens, or entire specimens if beyond bleeding stage, in
plastic bag and freeze. In case no method of freezing is available, icing
for a short period prior to freezing may be acceptable. Labeling of both
blood and carcass is important.
3. Controls - live specimens of the affected organisms should be obtained
from an area within the same body of water which had not been influenced
by the causative agent. Once obtained these specimens should be handled
in a like manner.
7.2.8.2 The number of individuals involved and the species affected should be
enumerated in some manner. At most these will be estimates. Depending on
the given situation such as area or distance involved and personnel available,
enumeration of fish kills may be approached in one of the following ways:
1. For large rivers, establish observers at a station or stations (e.g.,
bridges) and count the dead and/or dying fish for a specified period of
time, then project to total time involved.
2. For large rivers and lakes, traverse a measured distance of shoreline,
count the number and kinds of dead or dying fish. Project numbers
relative to total distance of kill.
3. For lakes and large ponds, count the number and species within measured
areas, and then project to total area involved.
4. For smaller streams one may walk the entire stretch involved and count
number of dead individuals by species.
7.2.9 Sampling Other Biota
7.2.9.1 Sampling of benthic organisms after the more urgent aspects of the
kill investigation has been completed can prove to be valuable relative to the
extent and cause of the kill. Benthic invertebrate communities are sampled to
determine whether this assemblage, the primary food source of many fishes, has
been affected. Also, since this general form of aquatic life is somewhat
sedentary by nature, release of deleterious materials to their environment
will kill much of the biota. By making a series of collections up and
downstream from the affected area, the affected stretch of stream may be
delineated when the benthic populations are compared to those organisms from
the control area. Also, the causative agent may be realized when the
specifics of the benthic population present are analyzed. Other aspects of
the biota which should be considered are the aquatic plants. In lakes and
ponds floating and rooted plants should be enumerated and identified. The
collection of plankton samples (river and lakes) should be taken in order to
determine possible toxicity from toxin-producing species and to determine the
degree of bloom, which in itself may cause fish kills because of diurnal
dissolved oxygen levels. Both aquatic plants and macroinvertebrates may be
fixed in a 10% formalin solution and preserved in 70% ethanol.
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7.2.9.2 When the material causing a kill is known, some of the above sample
collections may not be required. However, if the cause of a kill is unknown,
the above samples plus other specific samples, dictated by the type of
fishery, may be required.
7.2.9.3 Graphic evidence has a maximum effect on people involved in pollution
cases. Two basic types of graphic evidence are: (1) hand drawn maps of the
general and specific location of the kill, extent of kill, plankton bloom,
location of dead and dying fish, etc. (2) photograph (color and black-and-
white) showing dead fish, oil slicks, nasty looking water, sampling location,
etc. All graphic evidence should be carefully documented as recommended in
Subsection 7.2.5, Legal Aspects, and Meyer and Barclay, 1990).
7.2.9.4 The magnitude of a fish kill should be carefully documented. A
recognized method for enumerating the number and species which have been
killed should be selected and carefully followed so that data collected will
be admissible as evidence. Such methods are found in Meyer and Barclay (1990)
and references cited in Subsections 7.2.6.1 and 7.2.6.2.
7,2.10 Bioassays
7.2.10.1 Static bioassay techniques, as outlined in USEPA (1991), may be
effectively used to determine acute toxicity of wastes as well as receiving
waters. Toxicity testing can be done in-situ using live boxes, a mobile
bioassay laboratory, or the samples can be returned to a central laboratory
for testing.
7.2.11 Report
7.2.11.1 The final report should contain accurate information and should be
well organized to meet the requirements under Legal Aspects (Subsection?.2.5).
Essential elements of the report are: (1) introduction, (2) summary, (3)
description of the area, (4) description of all sampling methods and analyses,
(4) discussion of the magnitude of the fish kill and effects on other aquatic
organisms, (5) discussion of other water users in the affected area and (6)
conclusion. For additional recommendations, see the references listed in
Subsections 7.2.6.1, 7.2.6.2, and Section 12, Fisheries Bibliography, 12.7
Fish Kills.
7.2.12 Case History
7.2.12.1 A lower Mississippi River endrin-caused fish kill is an excellent
example of the investigation of a major fish kill Bartsch and Ingram (1966)
give the following summary (Table 9).
7.2.12.2 The investigation was designed to consider and eliminate potential
fish kill possibilities that were not involved and come to a point focus on
the real cause. It was found that the massive kills were not caused by
disease, heavy metals, organic phosphorus compounds, lack of dissolved oxygen
or unsuitable pH. Blood of dying river fish was found to have concentrations
of endrin equal to or greater than laboratory fish killed with this pesticide,
while living fish had lesser concentrations. Symptoms of both groups of dying
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TABLE 9. SUMMARY OF A LOWER MISSISSIPPI RIVER ENDRIN FISH KILL
INVESTIGATION1'2
I. Examination of usual environmental factors.
II. Elimination of parasites, bacterial or viral diseases, and
botulism as causes of mortalities1.
III. Consideration of toxic substances: Examination and
prognostication of symptoms of dying fish. Necropsy including:
Haematocrits and white cell counts
Brain tissue assay for organic phosphorus insecticide
Kidney tissue study
Tissue analysis for 19 potentially toxic metals
Gas chromatographic analysis of tissues, including blood, for
chlorinated hydrocarbon insecticides
IV. Exploration for toxic substances:
Bioassay with Mississippi River water
Bioassay with extracts from river bottom mud
Bioassay with tissue extracts from fish dying in river water and
bottom mud extracts
Bioassay with endrin to compare symptoms and tissue extract
analyses with those of dying fish in all bioassays.
V. Intensive chemical analysis for pesticides in the natural
environment, experimental environment, river fish, and
experimental animals.
VI. Surveillance of surface waters for geographic range and intensity
of pesticide contamination.
VII. Correlation and interrelation of findings.
Modified from Bartsch and Ingram (1966).
2The investigator should be aware of the fact that apparently healthy fish may
be harboring pathogenic bacteria in their bloodstreams (see Bullock and
Snieszko, 1969). Thus, there may be several factors involved in fish
mortalities, all of which may obscure the primary cause or causes.
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fish (river and bioassay) in the study (Table 9) were identical. It was
concluded from all data obtained that these fish kills were caused by endrin
poisoning.
7.3 Instream Flow Incremental Methodology (IFIM)
7.3.1 The IFIM was developed by Bovee (1982) for the U.S. Fish and Wildlife
Service and is widely utilized in the United States by the U.S. Fish and
Wildlife Service, state agencies, and consulting firms to estimate the effect
of change in instream flow on the habitat of stream fish and other aquatic
organisms (Baldridge and Amos, 1981; Gan and McMahon, 1990; Gore and Judy,
1981; Hilgert, 1982; Irvine et al., 1987; Mathur et al., 1985; Orth and
Maughan, 1982, 1986; Parsons and Hubert, 1988; Waite, 1989; Waite and
Barnhart, 1992). This methodology is only discussed here generally, but
investigators should consult the authors cited in this Subsection for their
application of fisheries bioassessment, management, and related research
needs.
7.3.2 The application of the IFIM and its effectiveness have been evaluated
and reviewed by several researchers (Bayha, 1978; Conder and Annear, 1987; Gan
and McMahon, 1990; Gore and Nestler, 1988; Irvine, et al., 1987; Mathur et
al., 1985; Orth and Maughan, 1982; 1986; Shirvell, 1989; Waite, 1989; Waite
and Barnhart, 1992). In addition, Wesche and Rechard (1980) reviewed and
summarized instream flow methods for fisheries and related research needs.
7.3.3 An important element of the IFIM is the use of physical habitat
simulation (PHABSIM) computer models (e.g., IFG-4, HABTAT) that relate changes
in discharge or stream channel structure to changes in the availability of
physical habitat (Waite and Barnhart, 1992). With PHABSIM the hydraulic and
physical variables of a stream or river are simulated for an assigned flow,
and the amount of usable habitat (weighted usable area or WUA) can be
predicted for a particular life stage of a particular species of fish. The
prediction of WUA is based on ecological data and on habitat use by selected
species of fish at various developmental life stages. The data are expressed
in terms of habitat utilization or probability of use curves (Bovee and
Cochnauer, 1977; Raleigh et al., 1984). The habitat utilization curves most
commonly used in the IFIM are those for current velocity, substrate particle
size, and water depth. According to Parsons and Hubert (1988), the values
that are generated by an IFIM study can be misleading if the habitat
utilization curves do not adequately reflect the conditions that fish of a
life stage need, prefer, or tolerate. In addition, the type of habitat used
by stream salmonids varies by species, life stage of the species, and
characteristics of the available habitat. Using data found in the literature
and additional research, Bovee (1978), Bovee and Cochnauer (1977), and Raleigh
et al. (1984) compiled and developed general standard habitat utilization
curves which could be broadly applied. Shirvell (1989) found that generic
curves were not always accurate. Waite and Barnhart (1992) developed habitat
utilization curves for allopatric fry and juveniles of steelhead Oncorhynchus
mykiss over a range of environmental conditions in a small stream with
moderate to high gradient, and also compared these curves with three standard
IFIM probability of use curves. Waite and Barnhart (1992) also concluded
that applying habitat utilization curves of one stream to generate WUA values
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for a different stream should be done only after the investigator has measured
and compared other stream characteristics, such as stream width, flow,
gradient, depth, substrate particle size, pool:riffle ratio, and seasonal
hydrography.
7.3.4 Recent studies by Layher and Brunson (1992) involve modification of the
habitat evaluation procedures for determining instream flow requirements in
warmwater streams; Olson-Rutz and Marlow (1992) studied the analysis and
interpretation of stream channel cross-sectional data and discussed stream
channel form and bank stability importance to the biotic community structure
of riparian ecosystems.
7.4 Fish Marking and Tagging Techniques (Mark-and-Recapture)
7.4.1 The marking and tagging of fish are important techniques utilized to
obtain information necessary for research and management. They are often used
to study individual fish or fish populations. Marking or Tagging studies can
give investigators data on estimates of biomass, stocking success, migrations,
behavior, age, mortality rates, etc. For a review and synthesis of the
different types of devices and techniques (e.g., external, internal,
electronic, genetic, chemical tags and marks, etc.), consult Lagler (1956,
1978), Wydoski and Emery (1983), Parker et al. (1990).
7.5 Literature Cited
APHA. 1992. Special-Purpose Toxicity Tests, 8-6. Investigation of fish
kills, pages 10-80. In: Standard methods for the examination of water
and wastewater. 18th Edition. Amer. Public Health Association,
Washington, DC.
Amer. Fish. Soc. 1982. Monetary values of freshwater fish and fish-kill
counting guidelines. Amer. Fish. Soc. Special Publ. No. 13, Bethesda,
MD.
Ardinger, G.R. 1990. Equipment needed for field assessment. In: F.P. Meyer
and L.A. Barclay (eds.). Field manual for the investigation of fish
kills. U.S. Dept. Interior, Fish and Wildlife Service, Resource
Publication 177, Washington, DC. pp. 87-89.
ASTM. 1992. Standard practice for evaluating an effluent for flavor
impairment to fish flesh. ASTM Designation: D 3969 - 96, pp. 22-27.
ASTM, Philadelphia, PA
Baldridge, J.E. and D. Amos. 1981. A technique for determining fish habitat
suitability criteria: a comparison between habitat utilization and
availability. Page 251-258. In: N.B. Armantrout (ed.). Acquisition
and utilization of aquatic habitat inventory information. Amer. Fish.
Soc. Western Division, Bethesda, MD.
Barclay, L.A. 1990a. How to ship samples. In: F.P. Meyer and L.A. Barclay
(eds.). Field manual for the investigation of fish kills. U.S. Dept.
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Interior, Fish and Wildlife Service, Resource Publication 177,
Washington, DC. pp. 71-74.
Barclay, L.A. 19905. Preparing the testimony. In: P.P. Meyer and L.A.
Barclay (eds.). Field manual for the investigation of fish kills. U.S.
Dept. Interior, Fish and Wildlife Service, Resource Publication 177,
Washington, DC. pp. 83-86.
Bartsch, A.F. and W.N. Ingram. 1966. Biological analysis of water pollution
in North america. International Verein Limnol. 16:786-800.
Bayha, K. 1978. Instream flow methodologies for regional and national
assessments. U.S. Fish and Wildlife Service Biological Services Program
FWS/OBS-78/61
Bovee, K.D. 1978. Probability of use criteria for the family Salmonidae.
U.S. Fish and Wildlife Service Biological Services Program FWS/OBS-
78/07.
Bovee, K.D. 1982. A guide to stream habitat analysis using the instream
flow incremental methodology. U.S. Fish and Wildlife Service Biological
Services Program FWS/OBS-82/26.
Bovee, K.D. and T. Cochnauer. 1977. Development and evaluation of weighted
criteria, probability-of-use curves for instream flow assessments:
fisheries. U.S. Fish and Wildlife Service Biological Services Program
FWS/OBS-77/63.
Bullock, G.L. and S.F. Snieszko. 1969. Bacteria in blood and kidney of
apparently healthy hatchery trout. Trans. Amer. Fish. Soc. 98:268-
271.
Burdick, G.E. 1965. Some problems in the determination of the cause of fish
kills. In: Biological Problems in Water Pollution. Pub!. No. 999-WP-
25, U.S. Public Health Serv., Washington, DC.
Conder, A.L. and T.C. Annear. 1987. Test of weighted usable area estimates
derived from a PHABSIM model for instream flow studies on a trout
streams. North Amer. J. Fish. Manage. 7:339-350.
Can, K. and T. McMahon. 1990. Variability of results from the use of PHABSIM
in estimating habitat area. Regulated Rivers: Research and Management
233-239.
Gore, J.A. and R.D. Judy, Jr. 1981. Predictive models of benthic
macroinvertebrate density for use in instream flow studies and regulated
flow management. Can. J. Fish Aquatic Sci. 38:1363-1370.
Gore, J.A. and J.M. Nestler. 1988. Instream flow studies in perspective.
Regulated Rivers: Research and Management 2:03-101.
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Herman, R.L. 1990. The role of infectious agents in fish kills. In: F.P.
Meyer and L.A, Barclay (eds.). Field manual for the investigation
of fish kills. U.S. Dept. Interior, Fish and Wildlife Service, Resource
Publication 177, Washington, DC. pp. 45-56.
Herman, R.L. and F.P. Meyer. 1990. Fish kills due to natural causes. In:
F.P. Meyer and L.A. Barclay (eds.). Field manual for the investigation
of fish kills. U.S. Dept. Interior, Fish and Wildlife Service, Resource
Publication 177, Washington, DC. pp. 41-44.
Hilgert, P. 1982. Evaluation of instream flow methodologies for fisheries in
Nebraska. Nebraska Game and Parks Commission, Technical series 10,
Lincoln.
Hill, D.M. 1983. Fish kill investigation procedures. In: Nielsen, L.A. and
D.L. Johnson (eds.). Fisheries Technique. American Fisheries Society,
Bethesda, MD. pp. 261-274.
Hunn, J.B. 1990. Planning. In: F.P. Meyer and L. A. Barclay (eds.). Field
manual for the investigation of fish kills. U.S. Dept. Interior, Fish
and Wildlife Service, Resource Publication 177, Washington, DC. pp. 6-
9.
Hunn, J.B. and R.A. Schnick. 1990. Toxic Substances. In: F.P. Meyer and L.
A. Barclay (eds.). Field manual for the investigation of fish kills.
U.S. Dept. Interior, Fish and Wildlife Service, Resource Publication
177, Washington, DC. pp. 19-40.
Irvine, J.R., 1.6. Jowett, and D. Scott. 1987. A test of the instream flow
incremental methodology for underyearling rainbow trout, Sal mo
gairdneri , in experimental New Zealand streams. New Zealand J. Marine
Freshwater Res. 21:35-40.
Lagler, K.F. 1956. Freshwater fishery biology. Second Edition. William C.
Brown Co., Dubuque, iowa. 421 pp.
Lagler, K.F. 1978. Capture, sampling and examination of fishes. Pages 7-47.
In: methods for assessment of fish production in freshwater. Blackwell
Sci. Publ., Oxford, England. IBP handbook No. 3.
Layher, W.G. and K.L. Brunson. 1992. A modification of the habitat
evaluation procedure for determining instream flow requirements in
warmwaters streams. North Amer. J. Fish Manage. 12(1)47-54.
Mathur, D., W.H. Bason, E.J. Purdy, Jr., and C.A. Silver. 1985. A critique
of the instream flow incremental methodology. Can.J. Fish. Aquatic Sci.
42:825-831.
Meyer, F.P. 1990a. Writing the report. In: F.P. Meyer and L.A. Barclay
(eds.). Field manual for the investigation of fish kills. U.S. Dept.
Interior, Fish and Wildlife Service, Resource Publication 177,
Washington, DC. pp. 75-82.
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Meyer. P.P. 1990b. Test your skill. In: F.P. Meyer and L.A. Barclay
(eds.). Field manual for the investigation of fish kills. U.S. Dept.
Interior, Fish and Wildlife Service, Resource Publication 177,
Washington, DC. pp. 90-97.
Meyer, P.P. and L.A. Barclay (eds.). 1990. Field manual for the
investigation of fish kills. U.S. Department of the Interior, Fish and
Wildlife Service, Resource Publication 177, Washington, DC. [Copies of
this document may be purchased from the National Technical Information
Service (NTIS), 5285 Port Royal Road, Springfield, VA 22161 or from the
Superintendent of Documents, U.S. Government Printing Office,
Washington, DC. Stock number 024-010-00685-4
Meyer, F.P. and R.L. Herman. 1990. Interpreting the scene. In: F.P. Meyer
and L.A. Barclay (eds.). Field manual for the investigation of fish
kills. U.S. Dept. Interior, Fish and Wildlife Service, Resource
Publication 177, Washington, DC. pp. 10-18.
Olson-Rutz, K.M. and C.B. Marlow. 1992. Analysis and interpretation of
stream channel cross-sectional data. North Amer. J. Manage. 12(1):55-
61.
Orth, D.J. and O.E. Maughan. 1982. Evaluation of the instream flow
incremental methodology. Trans. Amer. Fish. Soc. 111:413-445.
Orth, D.J. and O.E. Maughan. 1986. In defense of the instream flow
incremental methodology. Can. J. Fish Aquatic Sci. 43:1092-1093.
Parker, N.C., A.E. Giorgi, R.C. Heidinger, D.B. Jester, Jr., E.D. Prince, and
G.A. Winans (eds.). 1990. Fish-marking techniques. American Fisheries
Society Symposium 7, Bethesda, MD. 893 pp.
Parsons, B.G.M. and W.A. Hubert. 1988. Influence of habitat availability on
spawning site selection by kokanees in streams. North Amer. J. Fish.
Manage. 8:426-431.
Raleigh, R.F., T. Hickman, R.C. Solomon, and P.C. Nelson. 1984. Habitat
suitability information: rainbow trout. U.S. Fish Wildlife Service
Biological Services Program FWS/OBS-84/10.60.
Schnick, R.A. 1990a. Quality assurance and rules of evidence. In: F.P.
Meyer and L.A. Barclay (eds.). Field manual for the investigation
of fish kills. U.S. Dept. Interior, Fish and Wildlife Service, Resource
Publication 177, Washington, DC. pp. 57-62.
Schnick, R.A. 1990b. Where to send samples for analysis. In: F.P. Meyer and
L.A. Barclay (eds.). Field manual for the investigation of fish kills.
U.S. Dept. Interior, Fish and Wildlife Service, Resource Publication
177, Washington, DC. pp. 63-70.
Shirvell, C.S.
habitat.
1989. Ability of PHABSIM to predict chinook salmon spawning
Regulated rivers: Research and Management 3:277-2889.
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Smith, L.L., Jr, B.G. Andreson, W.A. Chipman, J.B, Lackery, O.L. Meehean, E.
Schneberger, W.A. Spoor, C.M. Tarzwell. 1956. Procedures for
investigation of fish kills. A guide for field reconnaissance and data
collection. Ohio River Valley Water Sanitary Commission (ORSANCO),
Cincinnati, OH.
Thomas, N. 1969. Flavor of Ohio River channel catfish (IctaTarus punctatus
Raf.). USEPA, Cincinnati, OH.
Tracy, H,B. and J.C. Bernhardt. 1972. Guidelines for evaluating fish kill
damages and computing fish kill damage claims in Washington state.
State of Washington, Dept. El. 46 pp.
TVA. 1968. Fish kill in Boone Reservoir. Tennessee Valley Authority, Water
Quality Branch, Chattanooga, TN.
U.S. Dept. Interior. 1970. Investigating fish mortalities. FWPCA Publ. No.
CWT-5. Also available from USGPO as No. 0-380-257.
USEPA. 1973. Freshwater biology and pollution ecology. Training Manual.
U.S. Environmental Protection Agency, Water Programs Operations,
Training Program, Cincinnati, OH. pp. 47-11.
USEPA. 1979a. Handbook for analytical quality control in water and
wastewater laboratories, EPA/600/4-79/019. Environmental Monitoring
and Support Laboratory, U.S. Environmental Protection Agency,
Cincinnati, OH.
USEPA. 1979b. Methods for chemical analysis of water and waste. EPA-600/4-
79/020. Environmental Monitoring and Support Laboratory, U.S.
Environmental Protection Agency, Cincinnati, OH. (revised March, 1983).
USEPA. 1980. Fish kills caused by pollution in 1977. EPA/400/4-80-004.
U.S. Environmental Protection Agency, Office of Water Planning and
Standards, Washington, DC.
USEPA. 1991. Methods for measuring the acute toxicity of effluents and
receiving waters to freshwater and marine organisms. C.I. Weber (ed.).
EPA-6QO/4-90-027. U.S. Environmental Protection Agency, Environmental
Monitoring Systems Laboratory, Cincinnati, OH 45268.
Waite, I.R. 1989. A comparison of site specific and generic instream flow
incremental methodology microhabitat criteria for rearing steelhead.
Master's Thesis. Humboldt State University, Arcata, CA.
Waite, I.R. and R.A. Barnhart. 1992. Habitat criteria for rearing steelhead:
A comparison of site-specific and standard curves for use in the
instream flow incremental methodology. North Amer. J. Fish. Manage.
12(l):40-46.
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Wesche, T.A. and P.A. Rechard. 1980, A summary of instream flow methods for
fisheries and related research needs. Eisenhower Consortium Bulletin 9,
Eisenhower Consortium for Western Environmental Forestry Research. 122
pp.
Wydoski, R. and L. Emery. 1983. Tagging and Marking. In: Nielsen, L.A. and
D.L. Johnson (eds.). Fisheries Techniques. American Fisheries Society,
Bethesda, MD. pp. 215-238.
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SECTION 8
FISH BIOASSESSMENT PROTOCOLS FOR USE IN STREAMS AND RIVERS1
8.1 Introduction
8.1.1 Two levels of fish bioassessment analyses are presented. Fish
Bioassessment I constitutes a questionnaire approach where local and State
fisheries experts are canvassed for existing data and information; Fish
Bioassessment II consists of collecting fish at selected sites for biosurvey
analyses. The data collected in Fish Bioassessment II is used in the Index of
Biotic Integrity (IBI) {Karr et al., 1986} and the Index of well-being (Iwb)
or composite index (Gammon, 1976, 1980; Gammon et al., 1981, 1988). This
section provides an overview of the IBI and Iwb and their conceptual
foundations. Effective use of the Fish Bioassessment II requires information
presented in Angemeier and Karr (1986), Karr et al. (1986) and Gammon (1980).
Sample field and data sheets are presented for guidance.
8.1.2 Pilot studies based on use of the fish biosurvey (Fish Bioassessment
II) have been published. An overview of two of these studies is presented in
Plafkin et al. (1989). Other studies by Bramblett and Fausch (1991), Hughes,
and Gammon (1987), Ohio EPA (1987b, 1987c. 1990a), Plafkin et al. (1989),
Schrader (1989), Simon (1990, 1991), Steedman (1988), Yoder et al. (1981), and
those states or agencies cited in Subsection 8.15 have applied the IBI and
Iwb, or the modified Iwb, to assess the effects of impacts in habitats of
different regions of North America.
8.1.3 Use of Fish in Biosurveys
8.1.3.1 The bioassessment techniques presented here focus on the evaluation
of water quality, habitat, and fish community parameters. The fish survey
protocols were based largely on Karr's IBI (Karr, 1981; Karr et al., 1986;
Miller et al., 1988b), which uses fish community structure to evaluate water
quality. The integration of functional and structural compositional metrics,
which forms the basis for the IBI is a common element to the fish
bioassessment approach.
8.1.3.2 Advantage of Using Fish
8.1.3.2.1 Fish are good indicators of long-term (several years) effects and
broad habitat conditions because they are relatively long-lived and contain
mobile elements (Karr et al., 1986). In additions many species are relatively
sedentary in summer (Gerking, 1959).
8.1.3.2.2 Fish communities generally include a range of species that are
representation of a variety of trophic levels (omnivores, herbivores,
insectivores, planktivores, piscivores). They tend to integrate effects of
1Adapted from Plafkin et al. (1989).
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lower trophic levels; thus, the fish community structure can present an
integrated picture of the environmental health of a stream or river.
8.1.3.2.3 Fish are at the top of the aquatic food chain and are consumed by
humans, making them important target assemblage for assessing contamination
and habitat alteration.
8.1.3.2.4 Fish are relatively easy to collect and identify to the species
level. Most specimens can be sorted and identified in the field and released
unharmed.
8.1.3.2.5 Environmental requirements of common fish are comparatively well
known.
8.1.3.2,6 Life history information is extensive for most species.
8.1.3.2.7 Information on fish distribution is commonly available.
8.1.3.2.8 Aquatic life uses (water quality standards) are typically
characterized in terms of fisheries (coldwater, coolwater, warmwater, sport,
forage, commerci al).
8.1.3.2.9 Monitoring fish communities provides direct evaluation of
"fishability", which emphasizes the importance of fish to anglers and
commercial fishermen.
8.1.3.2.10 Fish account for nearly half of the endangered vertebrate species
and subspecies in the United States.
8.1.4 Fish Community Consideration
8.1.4.1 Seasonal changes in the relative abundance of the fish community
primarily occur during reproductive periods and (for some species) the spring
and fall migratory periods. However, because larval fish sampling is not
recommended in this method, reproductive period changes in relative abundance
are not of primary importance.
8.1.4.2 Generally, the preferred sampling season is mid to late summer and
early fall, when stream and riverflows are moderate to low, and less variable
than during other seasons. Although some fish species are capable of
extensive migration, fish populations and individual fish tend to remain in
the same area during summer (Funk, 1957; Gerking, 1959; Cairns and Kaesler,
1971). The Ohio EPA (Rankin, 1987, personal communication) confirmed that few
species or individuals of a species in perennial streams migrate long
distances. Hill and Grossman (1987) found that the three dominant fish
species in a North Carolina stream had home ranges of 13 to 19 m over a period
of 18 months. Ross et al. (1985) and Matthews (1986) found that stream fish
assemblages were stable and persistent for 10 years, recovering rapidly from
droughts and floods indicating that large population fluctuations are unlikely
to occur in response to purely natural environmental phenomena. However,
comparison of data collected during different seasons is discouraged, as is
data collected during or immediately after major flow changes.
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8.1.5 Station Siting
8.1.5.1 Fish Bioassessment II includes the collection of biological samples
to assess the biotic integrity of a given site. To meaningfully evaluate
biological condition, sampling locations must be carefully selected to ensure
generally comparable habitats at each station. Unless comparable physical
habitat is sampled at all stations, community differences attributable to a
degraded habitat will be difficult to separate from those resulting from water
quality degradation. The availability of habitats at each sampling location
can be established during preliminary reconnaissance. In situations where
evaluations at several stations on a waterbody will be compared, the station
with the greatest habitat constraints (in terms of productive habitat
availability) should be noted. The station with the least number of
productive habitats available will often determine the type of habitat to be
sampled at all stations of comparison.
8.1.5.2 Locally modified sites, such as small impoundments and bridge areas,
should be avoided unless data are needed to assess the effects of these
structures. Sampling near the mouths of tributaries entering large
waterbodies should also be avoided since these areas will have habitat more
typical of the larger waterbody (Karr et al., 1986).
8.1.5.3 Although the specific bioassessment objective is an important
consideration in locating sampling stations, all assessments require a site-
specific control station or reference data from comparable sites within the
same region. A site-specific reference area or site (Ohio EPA, 1990b, 1991)
is generally thought to be most representative of "best attainable" conditions
for a particular waterbody. However, regional reference conditions may also
be desirable to allow evaluation on a larger geographic scale. Where
feasible, effects should be bracketed by establishing a series or network of
sampling stations at points of increasing distance from the impact source(s).
These stations will provide a basis for delineating impact and recovery zones
(these zones are not "reference stations").
8.1.5.4 Omernik (1987) and Omernik and Gallant (1988) have provided an
ecoregional framework for interpreting spatial patterns in state and national
data. The geographical framework is based on regional patterns in land-
surface form, soil types, potential natural vegetation, and land use, which
vary across the county. The use of ecoregions or similar approaches can
provide a geographic framework for more efficient management of aquatic
ecosystems and their components (Hughes, 1985; Hughes et al., 1982, 1986,
1987; Hughes and Larsen, 1988; Larsen et al., 1988). One method for
evaluating fish community composition is utilizing the ecoregion approach.
Another approach includes regional reference sites or control sites. The
application of the ecoregion versus the reference site approaches have been
documented (e.g., Larson et al., 1986; Ohio EPA, 1987b, 1989, 1990b; Rohm et
al, 1987; Whittier et al., 1988), but further studies are still needed to
determine the effectiveness of these approaches for other regions of North
America. In addition, investigations will be required to (1) delineate areas
that differ significantly in their innate biological potential, (2) locate
reference sites within each ecoregion that fully support aquatic life uses;
and (3) develop biological criteria (e.g., define optimal values for the
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metrics recommended) using data generated with the fish bioassessment II
protocol.
8.1.6 Importance of Habitat Assessment
8.1.6.1 The procedures for assessing habitat quality presented in this
Section are an integral component of the final evaluation of impairment. The
matrix used to assess habitat quality is based on key physical characteristics
of the waterbody and the surrounding land. All of the habitat parameters
evaluated are related to overall aquatic life use and are potential factors
which could contribute to a limitation of the aquatic biota in the waterbody.
8.1.6.2 Habitat, as affected by instream and surrounding topographical
features, can be a major determinant to aquatic community potential. Both the
quality and quantity of available habitat will affect the structure and
composition of resident biological communities. The effects of such
pertubations can be minimized by sampling similar habitats at all stations
being compared. However, when all stations are not physically comparable,
habitat characterization is particularly important for proper interpretation
of biosurvey results.
8.1.6.3 Where habitat quality is similar, detected impacts can be attributed
to water quality factors. However, where habitat quality differs
substantially from reference conditions, the question of use attainability and
physical habitat alteration/restoration must be addressed. Final conclusions
regarding the presence and degree of biological impairment should thus include
an evaluation of habitat quality to determine the extent that habitat may be a
limiting factor. The habitat characterization matrix included in the fish
bioasessment II methods provides an effective means of evaluating and
documenting habitat quality at each biosurvey station.
8.1.7 Fish Sampling Methodology (See, Section 4, Sample Collection for
Analysis of the Structure and Function of Fish Communities.)
8.1.7.1 Use of Electrofishing, Seining, and Rotenoning
8.1.7.1.1 Although various types of gear are routinely used to sample fish,
electrofishers, seines, and rotenone are the most commonly used for collection
in freshwater habitats. As detailed earlier each method has advantages and
disadvantages (Nielsen and Johnson, 1983; Hendricks et al., 1980). However,
electrofishing is recommended for most fish field surveys because of its
greater applicability and efficiency. Local conditions may require
consideration of seining and/or the use of rotenone as optional collection
methods. Advantages and disadvantages of each approach are presented below.
8.1.7.2 Advantages of Electrofishing
1. Electrofishing allows greater standardization of catch per unit of effort.
2. Electrofishing requires less time and manpower than some sampling methods
(e.g., use of ichthyocides, like rotenone) (Hendricks et al., 1980).
131
-------
3. Electrofishing is less selective than seining (although it is selective
towards size and species) (Hendricks et a!., 1980) (See disadvantage
number 2).
4. If properly used, adverse effects on fish are minimized.
5. Electrofishing is appropriate in a variety of habitats.
8.1.7.3 Disadvantages of Electrofishing
1. Sampling efficiency is affected by turbidity, conductivity, aquatic
vegetation, depth, etc,
2. Although less selective than seining, electrofishing also is size and
species selective. Effects of electrofishing increase with body size.
Species specific behavioral and anatomical differences also determine
vulnerability to electroshocking (Reynolds, 1983).
3. Electrofishing is a hazardous operation that can injure field personnel if
proper safety procedures are ignored.
8.1.7.4 Advantages of Seining
1. Seines are relatively inexpensive.
2. Seines are lightweight and are easily transported and stored.
3. Seine repair and maintenance are minimal and can be accomplished onsite.
4. Seine use is not restricted by water quality parameters.
5. Effects on the fish population are minimal because fish are collected
alive and are generally unharmed.
8,1.7.5 Disadvantages of Seining
1. Previous experience and skill, knowledge of fish habitats and behavior,
and sampling effort are probably more important in seining than in the use
of any other approaches (Hendricks et al,, 1980).
2. Seining sample effort and results are more variable than sampling with
electrofishing or rotenoning.
3. Seine use is generally restricted to slower water with smooth bottoms, and
is most effective in small streams or pools without litter cover or
debris.
4. Standardization of unit of effort to ensure data comparability is
difficult.
8.1.7.6 Advantages of Using Rotenone
132
-------
1. The effective use of rotenone is independent of habitat complexity.
2. Rotenoning provides greater standardization of unit of effort than
seining.
3. Rotenoning has the potential, if used effectively, to provide more
complete censuring of the fish population than seining or electrofishing.
8.1.7.7 Disadvantages of Using Rotenone
1, Use of rotenone is prohibited in many states.
2, Application and detoxification can be time and manpower intensive.
3. Effective use of rotenone is affected by temperature, light, dissolved
oxygen, alkalinity, and turbidity (Hendricks et al., 1980).
4. Rotenoning typically has a high environmental impact; concentration
miscalculations can produce substantial fish kills downstream of the study
site.
8.2 Sampling Representative Habitat
8.2.1 The sampling approach advocated in the Fish Bioassessment II optimizes
the conservation of manpower and resources by sampling areas of representative
habitat. The fish survey provides a representative estimate of the fish
community at all habitats within a sitef and a realistic sample of fish likely
to be encountered in the water body. When sampling large streams, rivers, or
waterbodies with complex habitats, a complete inventory of the entire reach is
not necessary for the level of assessment used in the Fish Bioassessment II.
The sampling area should be representative of the reach, incorporating
riffles, runs, and pools if these habitats are typical of the stream in
question. Although a sampling site with two riffles, two runs, and two pools
is preferable, at least one of each habitat type should be evaluated. Mid-
channel and wetland areas of large rivers, which are difficult to sample
effectively, may be avoided. Sampling effort may be concentrated in near-
shore habitats where most species will be collected. In doing so, some deep
water or wetland species may be under-sampled, however, the data should be
adequate for the objective of the Fish Bioassessment II method.
8.3 Fish Sample Processing and Enumeration
8.3.1 To ensure data comparability for assessing biological condition with
the Fish Bioassessment II, sample processing and species enumeration must be
standardized.
8.3.2 Processing of the fish biosurvey sample includes identification of all
individuals to species, weighing (if the Index of well-being (Iwb) or biomass
data are desired), and recording the incidence of external anomalies. It is
recommended that each fish be identified and counted. Subsamples of abundant
species may be weighed if live wells are unavailable. This is especially
important for warmwater sites, where handling mortality is highly probable.
133
-------
The data from the counted and weighed subsample is extrapolated for the total.
Ohio EPA (1987a) has reported that subsampling reduced potential error and
made the extra time required for individual weighing insignificant.
Procedural details for subsampling are presented in Ohio EPA, 1987c.
Determination of species trophic status is also necessary for some IBI
metrics. It should also be standard practice to collect fish Total Length
(TL) and Standard Length (SL) information.
8.4 Fish Environmental Tolerance Characterizations
8.4.1 Use of the Index of Biological Integrity (IBI) in the Fish
Bioassessment II requires classification of fish species in terms of
environmental tolerance. Responses of individual species to pollution will
vary regionally and in accordance with the type of pollutant. The tolerance
characterizations of selected midwestern and northwestern fish species are
presented in Table 1. Effective use of the tolerance characterization
approach requires an appropriate regional tolerance characterization system.
Regional modification or substitutions may be based upon regional fish
references, historical distribution records, objective assessment of a large
statewide database, and toxicological test data. Application of the IBI
approach in the southeastern and southwestern United States, and its
widespread use by water resource agencies may result in additional
modifications. Past modifications have been reported (Subsection 8.8, Miller
et al., 1988a) without changing the IBI's basic theoretical foundations.
8.5 Fish Biosurvey and Data Analysis
8.5.1 Bioassessment Technique
8.5.1.1 A biological assessment involves an integrated analysis of the
functional and structural components of the aquatic communities. These
functional and structural components are evaluated through the use of 12
metrics based on fish. The range of pollution sensitivity exhibited by each
metric differs among metrics (Figure 1); some are sensitive across a broad
range of biological conditions, others only to part of the range.
8.5.1.2 The 12 IBI metrics used in the Fish Bioassessment II method are based
on fish representing different sensitivities (Figure 2), For example,
municipal effluents typically affect total abundance and trophic structure
(Karr et al., 1986). Unusually low total abundance generally indicates a
toxicant effect. However, some nutrient-deficient environments support a
limited number of individuals or individual species, and an increase in
abundance may indicate organic enrichment. Bottom dwelling species (e.g.,
darters, sculpins) that depend upon benthic habitats for feeding and
reproduction are particularly sensitive to the effects of siltation and
benthic oxygen depletion (Kuehne and Barbour, 1983; Ohio EPA, 1987b) and are
good indicators of habitat degradation.
8.5.1.3 For the fish biosurvey and habitat assessment, scores are assigned to
each metric or parameter based on a decision matrix. In the case of habitat
assessment, evaluation of the quality of the parameter is based on visual
observation. The score assigned to each habitat parameter is a compilation of
134
-------
TABLE 1. TOLERANCE DESIGNATIONS, TROPHIC STATUS, AND NORTH AMERICAN
ENDEMICITY OF SELECTED FISH SPECIES3
WILLAMETTE SPECIES1
Salmonidae
Chinook salmon
Cutthroat trout
Mountain whitefish
Rainbow trout
Cyprinidae
Chiselmouth
Common carp
Goldfish
Leopard dace
Longnose dace
Northern squawfish
Peamouth
Redside shiner
Speckled dace
Catostomidae
Largescale sucker
Mountain sucker
Ictaluridae
Brown bullhead
Yellow bullhead
Percopsidae
Sand roller
Gasterosteidae
Threespine stickleback
Centrarchidae
Bluegill
Largemouth bass
Smallmouth bass
White crappie
Percidae
Yellow perch
Trophic Level
piscivore
insectivore
insectivore
insectivore
herbivore
omnivore
omnivore
insectivore
insectivore
piscivore
insectivore
insectivore
insectivore
omnivore
herbivore
insectivore
insectivore
insectivore
insectivore
insectivore
piscivore
piscivore
insectivore
insectivore
Tolerance
intolerant
intolerant
intolerant
intolerant
intermediate
tolerant
tolerant
intermediate
intermediate
tolerant
intermediate
intermediate
intermediate
tolerant
intermediate
tolerant
tolerant
tolerant
tolerant
intermediate
intermediate
Origin
native
native
native
native
native
exotic
exotic
native
native
native
native
native
native
native
native
introduced
introduced
intermediate native
intermediate native
introduced
introduced
introduced
native
intermediate native
aNot necessarily the final designations: designations may vary for different
regions.
Classifications for the Willamette River, Oregon were derived from Wydoski
and Whitney (1979). Moyle (1976), Scott and Grossman (1973), Simpson and
Wallace (1982), Dimick and Merryfield (1945), and Bond (1988, personal
communication.)
Classifications for midwestern fishes were taken from Karr et al. (1986) and
Ohio EPA (1987b).
Note: The information in this table is on going research and needs further
standardization.
135
-------
TABLE 1. TOLERANCE DESIGNATIONS, TROPHIC STATUS, AND NORTH AMERICAN
ENDEMICITY OF SELECTED FISH SPECIES (CONTINUED)
Cottidae
Paiute sculpin
Prickly sculpin
Reticulate sculpin
Torrent sculpin
MIDWEST SPECIES2
Petromyzontidae
Silver lamprey
Northern brook lamprey
Mountain brook lamprey
Ohio lamprey
Least brook lamprey
Sea lamprey
Polyodontidae
Paddlefish
Acipenseridae
Lake sturgeon
Shovel nose sturgeon
Lepisosteidae
Alligator gar
Shortnose gar
Spotted gar
Longnose gar
Amiidae
Bowfin
Hiodontidae
Goldeye
Mooneye
Clupeidae
Skipjack herring
Alewife
Gizzard shad
Threadfish shad
Salmonidae
Brown trout
Rainbow trout
Brook trout
Lake trout
Coho salmon
Chinook salmon
Lake herring
Lake whitefish
Osmeridae
Rainbow smelt
Trophic Level
insectivore
insectivore
insectivore
insectivore
piscivore
filterer
filterer
piscivore
filterer
piscivore
filterer
invertivore
invertivore
piscivore
piscivore
piscivore
piscivore
piscivore
insectivore
insectivore
piscivore
invertivore
omnivore
omnivore
insectivore
insectivore
insectivore
piscivore
piscivore
piscivore
piscivore
piscivore
invertivore
136
Tolerance
intolerant
intermediate
tolerant
intolerant
intermediate
intolerant
intolerant
intolerant
intermediate
intermediate
intolerant
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intolerant
intolerant
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
Origin
native
native
native
native
native
native
native
native
native
exotic
native
native
native
native
native
native
native
native
native
native
native
exotic
native
native
exotic
exotic
native
native
exotic
exotic
native
native
introduced
-------
TABLE 1. TOLERANCE DESIGNATIONS, TROPHIC STATUS, AND NORTH AMERICAN
ENDEMICITY OF SELECTED FISH SPECIES (CONTINUED)
Umbridae
Central mudminnow
Esocidae
Grass pickerel
Chain pickerel
Northern pike
Muskellunge
Cyprinidae
Common carp
Goldfish
Grass carp
Golden shiner
Hornyhead chub
River chub
Silver chub
Bigeye chub
Streamline chub
Gravel chub
Speckled chub
Blacknose dace
Longnose dace
Creek chub
Tonguetied minnow
Suckermouth minnow
Southern redbelly dace
Redside dace
Pugnose minnow
Emerald shiner
Silver shiner
Roseyface shiner
Redfin shiner
Rosefin shiner
Striped shiner
Common shiner
River shiner
Spottail shiner
Blackchin shiner
Bigeye shiner
Steelcolor shiner
Spotfish shiner
Bigmouth shiner
Sand shiner
Mimic shiner
Ghost shiner
Blacknose shiner
Pugnose shiner
Trophic Level
insectivore
piscivore
piscivore
piscivore
piscivore
omnivore
omnivore
herbivore
omnivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
general ist
insectivore
general ist
insectivore
insectivore
herbivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
To! erance
tolerant
intermediate
intermediate
intermediate
intermediate
tolerant
tolerant
intermediate
tolerant
intolerant
intolerant
intermediate
intolerant
intolerant
intermediate
intolerant
tolerant
intolerant
tolerant
intolerant
intermediate
intermediate
intolerant
intolerant
intermediate
intolerant
intolerant
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intolerant
intolerant
intermediate
intermediate
intermediate
intermediate
intolerant
intermediate
intolerant
intolerant
Origin
native
native
native
native
native
exotic
exotic
exotic
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
137
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TABLE 1. TOLERANCE DESIGNATIONS, TROPHIC STATUS, AND NORTH AMERICAN
ENDEMICITY OF SELECTED FISH SPECIES (CONTINUED)
Cyprinidae
Mississippi silvery minnow
Bullhead minnow
Bluntnose minnow
Fathead minnow
Central stoneroller
Popeye shiner
Silverjaw minnow
Central silvery minnow
Red shiner
Brassy minnow
Catostomidae
Blue sucker
Bigmouth buffalo
Black buffalo
Small mouth buffalo
Quilback
River carpsucker
Highfin carpsucker
Silver redhorse
Black redhorse
Golden redhorse
Shorthead redhorse
Greater redhorse
River redhorse
Harelip sucker
Northern hog sucker
White sucker
Longnose sucker
Spotted sucker
Lake chubsucker
Creek chubsucker
Ictaluridae
Blue catfish
Channel catfish
White catfish
Yellow bullhead
Brown bullhead
Black bullhead
Flathead catfish
Stonecat
Mountain madtom
Slender madtom
Freckled madtom
Northern madtom
Scioto madtom
Trophic Level
herbivore
omnivore
omnivore
omnivore
herbivore
insectivore
insectivore
herbivore
omnivore
omnivore
insectivore
insectivore
insectivore
insectivore
omnivore
omnivore
omnivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
invertivore
insectivore
omnivore
insectivore
insectivore
insectivore
insectivore
piscivore
general ist
insectivore
insectivore
insectivore
insectivore
piscivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
Tolerance
intermediate
intermediate
tolerant
tolerant
intolerant
intolerant
intermediate
intolerant
intermediate
intermediate
intolerant
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intolerant
intermediate
intermediate
intolerant
intolerant
intolerant
intolerant
tolerant
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
tolerant
tolerant
intermediate
intermediate
intolerant
intolerant
intolerant
intermediate
intolerant
intolerant
Origin
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
138
-------
TABLE 1. TOLERANCE DESIGNATIONS, TROPHIC STATUS, AND NORTH AMERICAN
ENDEMICITY OF SELECTED FISH SPECIES (CONTINUED)
Ictaluridae
Brindled madtom
Tadpole madtom
Anguill idae
American eel
Fundulidae
Western banded kill fish
Eastern banded kill fish
Blackstrip topminnow
Poeciliidae
Mosquitofish
Gadidae
Burbot
Moron idae
Trout -perch
Aphredoderidae
Pirate perch
Atherinidae
Brook silverside
Percichthyidae
White bass
Stripped bass
White perch
Yellow bass
Centrarchidae
White crappie
Black crappie
Rock bass
Small mouth bass
Spotted bass
Largemouth bass
Warmouth
Green sunfish
Bluegill
Orangespotted sunfish
Longear sunfish
Redear sunfish
Pumpkin seed
Percidae
Sauger
Walleye
Yellow perch
Dusky darter
Blackside darter
Longhead darter
Trophic Level
insectivore
insectivore
piscivore
insectivore
insectivore
insectivore
insectivore
piscivore
insectivore
insectivore
insectivore
piscivore
piscivore
piscivore
piscivore
invertivore
invertivore
piscivore
piscivore
piscivore
piscivore
invertivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
piscivore
piscivore
piscivore
insectivore
insectivore
insectivore
Tolerance
intolerant
intermediate
intermediate
intolerant
tolerant
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
tolerant
intermediate
intermediate
intolerant
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intolerant
Origin
native
native
native
native
native
native
exotic
native
native
native
native
exotic
exotic
exotic
exotic
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
139
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TABLE 1. TOLERANCE DESIGNATIONS, TROPHIC STATUS, AND NORTH AMERICAN
ENDEMICITY OF SELECTED FISH SPECIES (CONTINUED)
Percidae
Slenderhead darter
River darter
Channel darter
Gilt darter
Logperch
Crystal darter
Eastern sand darter
Western sand darter
Johnny darter
Greenside darter
Banded darter
Variegate darter
Spotted darter
Bluebreast darter
Tippecanoe darter
Iowa darter
Rainbow darter
Orangethroat darter
Fantail darter
Least darter
Slough darter
Sciaendiae
Freshwater drum
Cottidae
Spoonhead sculpin
Mottled sculpin
Slimy sculpin
Deepwater sculpin
Gasterosteidae
Brook stickleback
Trophic Level
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
invert ivore
insectivore
insectivore
insectivore
insectivore
insectivore
Tolerance
intolerant
intermediate
intolerant
intolerant
intermediate
intolerant
intolerant
intolerant
intermediate
intermediate
intolerant
intolerant
intolerant
intolerant
intolerant
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
Origin
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
native
140
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Assign
Identify Regional Fish Fauna
Species to Trophic, Tolerance, and Origin
Assess
D(
Guilds
Available Data for Metric Suitability anil Stream
Size Patterns
:ve)op Scoring Criteria from Reference Sit
es
Quantitative)!
• Sample Fish
List Abundances of Species, Hybrids, and Anomalies
Calculate and Score Metric Values
METRIC SCORES (IB!)
Metric
1 . Number of native fish
species
Scoring Criteria'*'
S 3 1
>67« 33-67% <33%,
2. Number of darter or bemhic species >67% 33-67$ <33% \^
3. Number of sunfish or
pool species >67% 33-67% <33% 7No1
4. Number of sucker or long-lived species >61% 33-67% <33% /
5. Number of intolerant
species
6. Proportion of green, sunfish or to
individuals
>67% 33-67% <33%
erant
<\i)% 10-25% >25%
7. Proportion omnisorous individuals <20% 20-45% >45%
8. Proportion insectivores >
9. Proportion top carnivores
10. Total number of individuals
1 i . Proportion hybrids or
exotics
459"r 20-45% <20%
>5% 1-5% <\%
>67% 33-67% <33%
12. Proportion with disease/anomalies
0% 0-1% >i%
5%
"''Metrics 1-5 art scored relative to the maximum species richness line.
Metric 10 is drawn from reference site data.
INDEX SCORE INTERPRETATION""
IBI Integrity Class
58-60 Excellent
Characteristics
Comparable to pristine conditions.
exceptional assemblage of species
48-52 Good
Decreased species richness,
intolerant species in particular;
sensitive species present
40-14 Fair
Intolerant and sensitive species
absent; skewed trophic structure
28-34 Poor
Top carnivor
es and many expected
species absent or rare; omnivores and
tolerant species dominant
12-22 Very Poor
Few species
and individuals present;
tolerant species dominant; diseased
fish frequent
'"From Karr « al. 1986; Ohio EPA
1987.
Recommendations
Not Universal
Criteria
Figure 1, Flowchart of biosurvey approach for Fish Bioassessment II
141
-------
a range of scores and is weighted in terms of its contribution to the total
habitat quality. The scores assigned to the fish metrics are based on
computed values of the metrics and a station comparison, wherein the regional
or stream reference station serves as the highest attainment criterion or
score for the area. Comparison of the total score computed for the metrics
orparameters with that of the reference station provides a judgment as to
impairment of biological condition.
8.5.1.4 The condition of the aquatic community needs to be evaluated and
interpreted within the context of habitat quality in order to determine
effects and likely causal factors. A poor habitat in terms of riparian
vegetation, bank stability, stream substrate, etc., would not be conducive to
supporting a well-developed community structure. The attainment of a higher
quality biological condition may be prohibited by the constraints of habitat
quality.
8.6 Fish Bioassessment I
8.6.1 The intent of the Fish Bioassessment I is to consist of a
questionnaire, to serve as a screening tool, and to maximize the use of
existing knowledge of fish communities. Note: The Fish Bioassessment I
method is not an option for a minimum state bioassessment program. The
Biological Condition
Metrics
Soecies
Darters
Sunfishes
Suckers
tntolerants
% Green Sunfisfi
% Grnnjvores
% Insectivorous Cyprinids
% Piscivores
IMumbei
% Hybrids
% Diseased
Non- Severely
Impaired Impaired
i f
L™ ,,__ „ _ _|
In, „ , J
ii.^ .,.™»J
Figure 2.
Range of sensitivities of biosurvey for Fish Bioassessment II
metrics in assessing biological condition (from Karr et al., 1986)
142
-------
questionnaire polls State fish biologists and university ichthyologists
believed knowledgeable about the fish assemblages in stream reaches of
concern. The questionnaire (Figure 3) is modeled after one used in a
successful national survey of 1,300 river reaches or segments (Judy et al.,
1984). Unlike field surveys, questionnaires can provide information about
tainting or fish tissue contamination and historical trends and conditions.
Disadvantages of the questionnaire approach include inaccuracy caused by hasty
responses, a desire to report conditions as better or worse than they are, and
insufficient knowledge. The questionnaire provides a qualitative assessment
of a large number of water bodies quickly and inexpensively. Its quality
depends on the survey design (the number and location of waterbodies), the
questions presented, and the knowledge and cooperation of the respondents.
8.6.2 This section provides guidance on the design and content of the
questionnaire survey. Judy et al. (1984) found that State fish and game
agencies have a vested interest in assuring the quality of the data, and they
generally provide reliable information.
8.6.3 Design of Fish Assemblage Questionnaire Survey
8.6.3.1 Selection of stream reaches requires considerable forethought. If
the survey program is statewide or regional in scope, a regional framework is
advisable. Regional reference reaches can be selected to serve as benchmarks
for comparisons (Hughes et al., 1986). These sites should be characteristic
of the water body types and sizes in the region and should be minimally
impacted. The definition of minimal impact varies from region to region, but
includes those waters that are generally free of point sources, channel
modification, and diversions, and have diverse habitats, complex bottom
substrate, considerable instream cover, and a wide buffer or natural riparian
vegetation.
8.6.3.2 Remaining sites should also be selected carefully. If the
questionnaire focuses on larger streams, a 1:1,000,000 scale topographic map
should be used for stream reach selection. Reaches of small streams should be
selected from the largest scale map possible; reaches selected from 1:250,000
versus 1:24,000 scale topographic maps may omit as much as 10 percent of the
permanent streams in humid, densely forested areas. Small, medium, and large
streams should be selected based on their importance in the region,
8.6.3.3 The potential respondent (or the agency chief if a number of agency
staff are to be questioned) should be contacted initially by telephone to
identify appropriate respondents. To ensure maximum response, the
questionnaire should be sent at times other than the field season and the
beginning and end of the a fiscal year or other seasonally busy time. The
questionnaire should be accompanied by a personalized cover letter written on
official stationary, and closed by an official title below the signature. A
stamped, self-addressed return envelope increases the response rate.
Materials mailed first or priority class are effective; special delivery and
certified letters are justified in follow-up mailings. Telephone contact is
advisable after three follow-up notes.
143
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FISH ASSEMBLAGE QUESTIONNAIRE
INTRODUCTION
This questionnaire is part of an effort to assess the biological health
or integrity ol the flowing waters of this state. Our principle focus is
on the biotic health of the designated waterbody as indicated by its fish
community. You were selected to participate in the study because of your
expertise in fish biology and your knowledge of the vmterbody identified
in this questionnaire.
Using the scale below, please circle the rank, (at left) corresponding to
the explanation (at right) that best describes your impression of the
condition of the vaterbody. Please complete all statements. If you feel
that you cannot complete the questionnaire, check here [ ] and return
it. If you are unable to complete the questionnaire but are avare of
someone who is familiar with the waterbody, please give this person's
name, address, and telephone number in the space provided below.
Waterbody code
Vaterbody name
Vaterbody location (also see nap)
State County Long/Lat
Ecoregion
Vaterbody size
Stream (<1 cfs, 1-10 cfs, >10 cfs)
(Answer questions 1-4 using the scale below.)
5 Species composition, age classes, and trophic structure comparable to
non (or minimally) impacted sites of similar waterbody size in that
ecoregion.
4 Species richness somewhat reduced by loss of some intolerant species;
young of the year of top carnivores rare; less than optimal
abundances, age distributions, and trophic structure for vaterbody
size and ecoregion.
3 Intolerant species absent, considerably fever species and individuals
than expected for that waterbody size and ecoregion, older age classes
of top carnivores rare, trophic structure skewed toward omnivory.
Figure 3. Fish assemblage questionnaire for use with Fish Bioassessment I.
144
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2 Dominated by highly tolerant species, omnlvpres, and habitat
generalists; top carnivores rare or absent; older age classes of all
but tolerant species rare; diseased £ish and anomalies relatively
common for that vaterbody size and ecoregion.
1 Few individuals and species present, mostly tolerant species and small
individuals! diseased fish and anomalies abundant compared to other
similar-sized vaterbodies in the ecoregion.
0 No fish
(Circle one number using the scale above.)
1, Rank the current conditions of the reach
543210
2. Rank the conditions at the reach 10 years ago
543210
3. Given present trends, hov vlll the reach rank 10 years £ron nov?
5 4 3 2.1 0
4. If the major human-caused limiting factors vere eliminated, hou
vould the reach rank 10 years from nov?
(Complete each subsection by circling the single «ost appropriate
limiting factor and probable cause.)
Subsection 1—Water Quality
Limiting factor
5 Temperature too high
6 Temperature too lov
7 Turbidity
8 Salinity
9 Dissolved oxygen
1C Gas supersaturation
11 pH too acidic
12 pH too basic
13 Nutrient deficiency
14 Nutrient surplus
15 Toxic substances
16 Other (specify belov)
17 Not limiting
Probable causa
18 Primarily upstream
19 Bithin reach
20 Point source discharge
21 Industrial
22 Municipal
23 Combined sever
24 Hitting
25 Dam release
26 Nonpoint source discharge
27 Individual sevage
28 Urban runoff
29
30
31
32
33
34
35
36 Natural
37 Unknown
38 Other (specify belov)
Landfill leachate
Construction
Agriculture
Feedlot
Grazing
Silviculture
Mining
Figure 3. Fish assemblage questionnaire for use with Fish Bioassessment I
(Continued),
145
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Subsection 2—Water Quantity
Limiting factor
39 Below optimum flows
40 Above optimum flows
41 Loss of flushing flows
42 Excessive flow fluctuation
43 Other (specify below)
44 Not limiting
Probable source
45 Dam
46 Diversion
47 Watershed conversion
48 Agriculture
49 Silviculture
50 Grazing
51 Urbanization
52 Hining
53 Natural
54 Unknown
55 Other (specify below)
Subsection 3--Habitat Structure
Limiting factor
56 Excessive siltation
5? Insufficient pools
58 Insufficient riffles
59 Insufficient shallows
60 Insufficient concealment
61 Insufficient reproductive
habi tat
62 Other (specify below)
63 Not limiting
Subsection 4 — Fish Community
Limiting factor
76 Qverharvest
77 Underharvest
78 Fish stocking
79 Non-native species
80 Migration barrier
81 Tainting
82 Other (specify below)
83 Not limiting
Probable cause
64 Agriculture
65 Silviculture
66 Hining
67 Grazing
68 Dam
69 Diversion
70 Channelization
71 Snagging
72 Other channel modifications
73 Natural
74 Unknown
75 Other (specify below)
Probable source
84 Fishermen
85 Aquarists
86 State agency
87 Federal agency
88 Point source
89 Nonpoint source
90 Natural
91 Unknown
92 Other (specify below)
Subsection _5—Hajor Limiting Factor
93 Water quality
94 Water quantity
95 Habitat structure
96 Fish community
97 Other (specify)
Your name (please print)
Figure 3. Fish assemblage questionnaire for use with Fish Bioassessment I
(Continued),
146
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8.6.4 Response Analysis
8.6.4.1 Questionnaire response should provide the following information:
1. The integrity of the fish community
2. The frequency of occurrence of particular limiting factors and causes
3. The frequency of occurrence of particular fish community condition
characterizations for the past, present, and future
4. The geographic patterns in these variables
5. The temporal trends in the variables
6. Effect of water body type and size on the spatial and temporal trends and
the associated limiting factors
7. The likelihood of improvement and degradation
8. The major limiting factor
8.6.4.2 The questionnaire data are most effectively analyzed by using a
microcomputer and an interactive data base management software (e.g., dBase
III or Revelation). This software reduces data entry errors and facilitates
the qualitative analysis of numerous variables. Results can be reported as
histograms, pie graphs, or box plots. If such a system is unavailable data
can be analyzed and the results plotted by hand.
8.7 Fish Bioassessment II
8.7.1 Introduction
8.7.1.1 Fish Bioassessment II involves careful, standardized field
collection, species identification and enumeration, and community analyses
using biological indices or quantification of the biomass and numbers of key
species. The Fish Bioassessment II survey yields an objective, discrete
measure of the health of the fish community that usually can be completed
onsite by qualified fish biologists (difficult species identifications may
require laboratory confirmation). Data provided by the Fish Bioassessment II
can allow assessment to use attainment, can be used to develop biological
criteria, prioritize sites for further evaluation, provide a reproducible
impact assessment, and be used to monitor trends in fish community status.
Fish Bioassessment II is based primarily on the Index of Biotic Integrity
(IBI) by Karr (1981). A more detailed description of this approach is
presented in Karr et al. (1986) and Ohio EPA (1987b). Regional modification
and applications are described in Hughes and Gammon (1987), Leonard and Orth
(1986), Lyons (1992), Steedman (1988), Wade and Stalcup (1987), Miller et al.
(1988a), and Simon (1990, 1991).
147
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8.7.2 Field Survey Methods
8.7.2.1 Fish Bioassessment II involves field evaluation of both
physical/chemical and habitat characteristics (see Subsection 8.13, Figures 9,
10, and 11), an impairment assessment (Figure 4), and a fish community
biosurvey. Because it provides critical information for evaluating the cause
and source of impairment, the habitat and physical characterization are
essential to Fish Bioassessment II. The approach for conducting the Fish
Bioassessment II site-specific fish community analysis is based on the use of
the IBI (Figure 1).
8.7.3 Sample Collection
8.7.3.1 Electrofishing, the most common technique used by agencies that
monitor fish communities, and the most widely applicable approach for stream
habitats, is the sampling technique recommended for use with the Fish
Bioassessment II. However, pilot studies may indicate the need for different
or multiple techniques and gear found in this document.
8.7.3.2 The fish community biosurvey data are designed to be representative
of the fish community at all station habitats, similar to the "representative
qualitative sample" proposed by Hocutt (1981). The sampling station should be
representative of the reach, incorporating at least one (preferably two)
riffle(s), run(s), and pool(s) if these habitats are typical of the stream in
question. Sampling of most species is most effective near shore and cover
(Macrophytes, boulders, snags, brush). The biosurvey is not an exhaustive
inventory, but it provides a realistic sample of fishes likely to be
encountered in the waterbody. Sampling procedures effective for large rivers
are described in Gammon (1980), Hughes and Gammon (1987), and Ohio EPA
(1987b).
8.7.3.3 Typical sampling station lengths range from 100-200 meters for small
streams to 500-1000 meters in rivers, but are best determined by pilot
studies. The size of the reference station should be sufficient to produce
100-1000 individuals and 80-90 percent of the species expected from a 50
percent increase in sampling distance. Sample collection is usually done
during the day, but night sampling can be more effective if the water is
especially clear and there is little cover (Reynolds, 1983; Sanders, 1991;
Sanders, 1992). Use of block nets set (with as little wading as possible) at
both ends of the reach increases sampling efficiency for large, mobile species
sampled in small streams.
8.7,3.4 The community-level assessment of fish assemblages using the Fish
Bioassessment II requires that all fish species (not just gamefish) be
collected. This reduces the effects of stocking and fishing and acknowledges
the growing public interest in nongame species. Small fish that require
special gear for their effective collection may be excluded. Exclusion of
young-of-the-year fish during collection can have a minor effect on IBI scores
(Angermeier and Karr, 1986), but lowers sampling costs and reduces the need
for laboratory identification. Karr et al., (1986) recommended exclusion of
fish less than 20 mm in length. This recommendation should be considered on a
regional basis and is also applicable to large fish requiring special gear for
148
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IMPAIRMENT ASSESSMENT SHEET
1. Detection of impairment: Impairment detected No impairment
(Complete Items 2-6) detected
(Stop here)
2, Biological impairment indicator:
Pish Other aquatic communities
s«nsitive species reduced/absent ^ M«croinvert*brataa
dominance of tolerant species ^ Ptriphyton
skeved trophic structure ___ Hacrophytes
abundance reduced/unusually hign
^~ biomass reduced/unusually high
hybrid or exotic abundance
unusually high
poor size class representation
high incidence of anomalies
3. Brief description of problem:
Tear and date of previous surveys;
Survey data available in:
4, Cause (indicate major cause); organic enrichment toxicants flow
sediment temperature poor habitat
other
5. Estimated areal extent of problem (a ) and length of stream reach
affected (a) vhere applicable:
6, Suspected source{s) of problem
point source mine
urban runoff dam or diversion
agricultural runoff channelization or snagging
silvicultural runoff natural
livestock other
landfill unknown
Comments:
Figure 4. Impairment assessment sheet for use with Fish Bioassessment II
149
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collection (e.g., sturgeon). The intent of the sample (as with the entire
Fish Bioassessment II method) is to obtain a representative estimate of the
species present, and their abundances, in a reasonable amount of effort.
8.7.3.5 Sampling effort among stations is standardized as much as possible.
Regardless of the gear used, the collection method, site length (or area), and
work hours expended must be comparable to allow comparison of fish community
status among sites. Major habitat types (riffle, run, and pool) sampled at
each site and the proportion of each habitat type sampled should also be
comparable. Generally 1 to 2 hours of actual sampling time are required, but
this varies considerably with the gear used and the size and complexity of the
site.
8.7.3.6 Atypical conditions, such as high flow, excessive turbidity or
turbulence, heavy rain, drifting leaves, or other unusual conditions that
affect sampling efficiency, should be avoided.
8.7.3.7 Glare, a frequent problem, is reduced by wearing polarized glasses
during sample collection.
8.7.3.8 At least four individuals (one with the electrofisher, two fish
netters, and one for holding container of collected fish) are necessary for
effective electrofishing, and electrofishing efficiency is increased by having
experienced netters involved.
8.7.4 Sample Processing
8.7.4.1 A field collection data sheet (Figure 5) is completed for each
sample. Sampling duration and area or distance sampled are recorded in order
to determine level of effort. Species may be separated into adults and
juveniles by size and coloration; then total numbers and weights and the
incidence of external anomalies are recorded for each group. Reference
specimens of each species from each site are preserved in 10 percent
formaldehyde (see Section 5, Fish Specimen Processing), the jar labeled, and
the collection placed with the State ichthyological museum to confirm
identifications and to constitute a biological record. This is especially
important for uncommon species, for species requiring laboratory
identification, and for documenting new distribution records. If retained in
a live well, most fish can be identified, counted, and weighted in the field
by trained personnel and returned to the stream alive. In warmwater sites,
where handling mortality is highly probable, each fish is identified and
counted, but for abundant species, subsampling may be considered. When
subsampling is employed, the subsample is extrapolated to obtain a final
value. Subsampling for weight is a simple, straightforward procedure, but
failure to examine all fish to determine frequency of anomalies (which may
occur in about 1 percent of all specimens) can bias results. The trade off
between handling mortality and data bias must be considered on a case-by-case
basis. If a site is to be sampled repeatedly over several months (i-e->
monitoring ), the effect of sampling mortality may outweigh data bias.
Holding fish in live boxes in shaded, circulating water will substantially
reduce handling mortality. More information on field methods is presented in
Karr et al. (1986) and Ohio EPA (1987a, 1987b, 1989).
150
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Collection No.
Page
of
State or Country
Locality
FISH FIELD COLLECTION DATA SHEET
County
Date
Water
Vegetation
Sampling duration (min.)
Bottom
Shore
Distance from shore or stream width _
Habitat complexity/quality (excellent
Sampling distance (m)
Depth of capture
Method of capture
Collected by
Temp
Air
good
fair
Orig. preservation
Weather
Gear/crew performance
Comments
Sampling area (m)
poor
very poor
Date
number of individuals
number of anomalis*
Flow (flood bankfull moderate low)
Genus/Species
Adults
No.
Wt.
Juvenjles
No. Wt.
Anomalies(*)
No.
*Discoloration, deformities, eroded fins, excessive mucus, excessive external
parasites, fungus, poor condition, reddening, tumors, and ulcers.
Figure 5, Fish field collection data sheet for use with Fish Bioassessment
II.
151
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8.7.5 Data Analysis Techniques
8.7.5.1 Based on observations made in the assessment of habitat, water
quality, physical characteristics, and the fish biosurvey, the investigator
concludes whether impairment is detected. If impairment is detected, the
probable cause and source is estimated and recorded on an Impairment
Assessment Sheet (Figure 4). A preliminary judgment on the presence of
biological impairment is particularly important if the Fish Bioassessment I is
not used prior to the Fish Bioassessment II.
8.7.5.2 Data can be analyzed using the Index of Biotic Integrity (IBI) (or
individual IBI metrics), the Index of well-being (Iwb) (Gammon, 1976, 1980),
and multivariate statistical techniques to determine community similarities.
Detrended correspondence analysis (DCA) is a useful multivariate analysis
technique for revealing regional community patterns and patterns among
multiple sites (Matthews et al., 1992). It also demonstrates assemblages with
compositions differing from others in the region or reach. The reader may
consult Gauch (1982) and Hill (1979) for descriptions of, and software for,
OCA. Data analyses and reporting, including parts of the IBI, can be computer
generated. Computerization reduces the time needed to produce a report and
increases staff capability to examine data patterns and implications. The
Illinois EPA has developed software to assist the professional aquatic
biologists in calculating IBI values in Illinois streams (Bickers et al.
1988). Use of this software outside Illinois or the particular ecoregion
without modification is not recommended. However, hand calculation in the
initial use of the IBI promotes understanding of the approach and provides
insight into local inconsistencies.
8.7.5.3 The IBI is a broadly-based index firmly grounded in fisheries
community ecology (Karr, 1981; Karr et al., 1986), The IBI incorporates
zoogeographic, ecosystem, community, population, and individual perspectives.
It can accommodate natural differences in the distribution and abundance of
species that result from differences in waterbody size, type, and region of
occurrence (Miller et al., 1988a). Use of the IBI allows national comparisons
of biological integrity without the traditional bias for small coldwater
streams (e.g., a salmon river in Alaska and a minnow stream in Georgia both
could be rated excellent if they were comparable to the best streams expected
in their respective regions).
8.7.5.4 Karr et al. (1986) provided a consistent theoretical framework for
analyzing fish community data. The IBI uses 12 biological metrics to assess
integrity based on the fish community's taxonomic and trophic composition and
the abundance and condition of fish. Such multiple-parameter indices are
necessary for making objective evaluations of complex systems. The IBI was
designed to evaluate the quality of small mid-western streams but has been
modified for use in many regions of the country and in large rivers
(Subsection 8.8).
8.7.5.5 The metrics attempt to quantify an ichthyologist's best professional
judgment of the quality of the fish community. The IBI utilizes professional
judgment, but in a prescribed manner, and it includes quantitative standards
for discriminating fish community condition. Judgment is involved in choosing
152
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the most appropriate population or community element that is representative of
each metric and in setting the scoring criteria. This process can be easily
and clearly modified, as opposed to judgments that occur after results are
calculated. Each metric is scored against criteria based on expectation
developed from appropriate regional reference sites. Metric values
approximating, deviating slightly from, or deviating greatly from values
occurring at the reference sites are scored as 5, 3, or 1, respectively. The
scores of the 12 metrics are added for each station to give an IBI of 60
(excellent) to 12 (very poor). Trophic and tolerance classifications of
midwestern and northwestern fish species are listed in Table 1. Additional
classifications can be derived from information in State and regional fish
texts or by objectively assessing a large statewide database. Use of the IBI
in the southern and southwestern United States and its widespread use by water
resource agencies may result in further modifications. Past modifications
have occurred (Subsection 8,8; Miller et al., 1988a) without changing the
IBI's basic theoretical foundations. Sample calculations of the IBI are given
in Plafkin et al. (1989).
8.7,6 The steps in calculating the IBI (Figure 1) are explained below:
8.7.6.1 Assign species to trophic guilds; identify and assign species
tolerances. Where published data are lacking, assignments are made based on
knowledge of closely related species and morphology.
8.7.6.2 Develop scoring criteria for each IBI metric. Maximum species
richness (or density) lines are developed from a reference database.
8.7.6.3 Conduct field study and identify fish; note anomalies, eroded fins,
poor condition, excessive mucous, fungus, external parasites, reddening,
lesions, and tumors. Complete field data sheets (Figure 5).
8.7.6.4 Enumerate and tabulate number of fish species and relative
abundances.
8,7.6.5 Summarize site information for each IBI metric.
8.7.6.6 Rate each IBI metric and calculate total IBI score.
8.7.6.7 Translate total IBI score to one of the five integrity classes.
8,7,6.8 Interpret data in the context of the habitat assessment (for a
discussion of Integration of Habitat, Water Quality, and Biosurvey data, see
Plafkin et al., 1989). Individual metric analysis may be necessary to
ascertain specific trends.
8.7.7 The Index
of the metrics.
variables to aid
dominant species
requirements and
provide examples
of Biotic Integrity (IBI) is based on an integrated analysis
However, individual IBI metrics may serve as separate
in data interpretation. Comparison of commonly-occurring and
are revealing, especially when related to their ecological
tolerances. Larsen et al. (1986) and Rchm et al« (1987)
of such regional characterizations of common and abundant
species. The Index of well-being (Iwb), (Gammon, 1980; Hughes and Gammon,
153
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1987) incorporates two abundance and two diversity estimates in approximately
equal fashion, thereby representing fish assemblage quality more realistically
than a single diversity or abundance measure. The Iwb is calculated as
Iwb = 0,5 In N+0.5 In B+H' +H'
N B
where N equals the number of individuals caught per kilometer, B equals the
biomass of individuals caught per kilometer, and H' is the Shannon diversity
index. Ohio EPA (1987b) and Gammon (1989, personal communication) found that
subtracting highly tolerant species from the number and biomass variables, or
modified Index of well-being (Iwb), increases sensitivity of the index in
degraded environments (Ohio EPA, 1987b; Yoder et al., 1981). The modified Iwb
has the same computational formula as the proposed Iwb by Gammon (1976). The
main difference is that any of 13 highly tolerant species, exotics, and
hybrids are deleted from the numbers and biomass components of the Iwb. The
tolerant and exotic species, however, are included in the two Shannon index
calculations. This modification eliminated the undesired effect caused by
high abundance of tolerant species, but retains the desired influence of the
Shannon indices (Ohio EPA, 1987b).
8.7.8 If the size of a particular fish population (e.g., trout or bass
species) is of concern, it can be estimated with known confidence limits by
several methods. One of the most popular approaches is the removal method
(Seber, 1982; Seber and LeCren, 1967; Seber and Whale, 1970). The approach
assumes a closed population, equal probability of capture for all fish, and a
constant probability of capture from sample to sample (equal sampling effort
and conditions). The removal method is applicable to situations in which the
total catch is large relative to the total population. If subsequent samples
produce equal or greater numbers than previous samples, the population must be
resampled. Population size in the two sample cases is
estimated by
N = C11/(C1 - C2)
where C1 and C2 are the number of fish captured in the first and second
samples, respectively. In the three sample cases, population size is
estimated by
N = 6X2 - 3XY - Y2+6XY - 3X2)1/2
18(X - Y)
where X = 2C, + C2, and Y = C2, + C3.
8.7.9 Many methods are available to calculate population statistics from
removal data including regression, maximum likelihood, and maximum weighted-
likelihood. Pubic-domain software is available to assist in calculating these
and other fisheries population statistics (Van Deventer and Platts, 1989).
8.8 Description of IB! Metrics
8.8.1 The IBI serves as an integrated analysis because individual metrics may
differ in their relative sensitivity to various levels of biological
condition. A description and brief rationale for each of the 12 IBI metrics
154
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is outlined below. The original metrics described by Karr (1981) for Illinois
streams (underlined) are followed by substitutes used in or proposed for
different geographic regions and stream sizes. Because of zoogeographic
differences, dissimilar families or species are evaluated in different
regions, with regional substitutes occupying the same general habitat or
niche. The sources for each substitute is footnoted below. Table 2 presents
an overview of the IBI metric variations for six areas of the United States
and Canada and their sources. Scoring criteria for the 12 original IBI
metrics (Karr, 1986) are included in Figure 1).
8.8.2 These metrics assess the species richness component of diversity and
the health of the major taxonomic groups and habitat guilds of fishes. Two of
the metrics assess community composition in terms of tolerant or intolerant
species. Scoring for the first five of these metrics or their substitute
metrics requires development of species-waterbody size relationships for
different zoogeographic regions. Development of this relationship requires
data sufficient to plot the number of species collected from regional
reference sites of various stream sizes against a measure of stream size
(watershed area, stream order) of those sites. A line is then drawn with
slope fit by eye to include 95 percent of the points. Finally the area under
the line is trisected into areas that are scored as 5, 3, or 1 (Figure 6). A
detailed description of these methods can be found in Fausch et al. (1984),
Ohio EPA (1987b), and Karr et al. (1986).
8.8.2.1 Metric 1. Total number of fish species (1,4,5). Substitutes: Total
number of native fish species (2,8), and salmonid age classes (6). This
number decreases with increased degradation; hybrids and introduced species
are not included. In coldwater streams supporting few fish species, the age
classes of the species found represent the suitability of the system for
spawning and rearing. The number of species is strongly affected by stream
size at small stream sites, but not at large river sites (Karr et al., 1986;
Ohio EPA, 1987b). Thus, scoring depends on developing species/waterbody size
relationships.
8.8.2.2 Metric 2. Number and identity (Page, 1983) of darter species (1).
Substitutes: Number and identity of sculpin species (2,4), benthic
insectivore species (3,4) salmonid yearlings (individuals) (6); number of
sculpins (individuals) (4); percent round-bodied suckers (5), sculpin, and
darter species (8). These species are sensitive to degradation resulting from
siltation and benthic oxygen depletion because they feed and reproduce in
benthic habitats (Kuehne and Barbour, 1983; Ohio EPA, 1987b). Many smaller
species live within the rubble interstices, are weak swimmers, and spend their
entire lives in an area of 100-400 m2 (Hill and Grossman, 1987; Matthews,
1986). Darters are appropriate in most Mississippi Basin streams; sculpins
and yearling trout occupy the same niche in western streams. Benthic
insectivores and sculpins or darters are used in small Atlantic slope streams
that have few sculpins or darters and round-bodied suckers are suitable in
large midwestern rivers. Scoring requires development of species/waterbody
size relationships.
8.8.2.3 Metric 3. Number and identity of sunfish species (1). Substitutes:
Number and identity of cyprinid species (2,4), water column species (3,4),
155
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TABLE 2, REGIONAL VARIATIONS OF IBI METRICS
en
CTl
Variations in IBI Metrics
New Central Colorado Western Sacramento-
Midwest England Ontario Appalachia Front Range Oregon San Joaquin
1. Total Number of Species
# native fish species
# salmonid age classes2
XX XX
X X
X
X
X
2. Number of Darter Species
I sculpin species
I benthic insectivore species
I darter and sculpin species
#salmonid yearlings (individuals)
% round-bodied suckers
Isculpins (individuals)
X
X
X
X
3. Number of Sunfish Species
I cyprinid species X
# water column species X
# sunfish and trout species X
# salmonid species X
# headwater species X
1Taken from Karr et al. (1986), Hughes and Gammon (1987), Miller et al. (1988a), Miller et al. (1988b), Ohio EPA
(1987b), and Steedman (1988).
2Metric suggested by Moyle (1976) or Hughes (1985) as a provisional replacement metric in small western salmonid
streams.
3An optional metric found to be valuable by Hughes and Gammon (1987).
Note: X = metric used in the region. Many of these variations are applicable elsewhere.
-------
TABLE 2. REGIONAL VARIATIONS OF IBI METRICS (CONTINUED)
4.
5.
en
6.
Variations in IBI Metrics
Number of Sucker Species
# adult trout species2
# minnow species
# sucker and catfish species
Number of Intolerant Species
# sensitive species
# amphibian species
presence of brook trout
% Green Sunfish
Midwest
X
X
X
X
X
New Central Colorado Western
England Ontario Appalachia Front Range Oregon
X X
X
X
X
X XX
X
Sacramento-
San Joaquin
X
X
% common carp
% white sucker
% tolerant species
% creek chub
% dace species
7. % Omnivores
% yearling salmonids2
8. % Insectivorous Cyprinids
% insectivores
% specialized insectivores
# juvenile trout
% insectivorous species
X
X
X
X
X
X
-------
TABLE 2. REGIONAL VARIATIONS OF IBI METRICS (CONTINUED)
9.
10.
11.
j
i
>
Variations in IBI Metrics
% Top Carnivores
% catchable salmonids
% catchable wild trout
% pioneering species
Density catchable wild trout
Number of Individuals
Density of individuals
% Hybrids
% introduced species
% simple lithophils
% simple lithophilic species
% native species
% native wild individuals
Midwest
X
X
X
X
X
X
New Central Colorado Western Sacramento-
England Ontario Appalachia Front Range Oregon San Joaquin
X X
X
X
X
X X X X X
X
X
X X
X
X
12. % Diseased Individuals
13. Total Fish Biomass3
X
X
-------
Log Watershed Area (mile
Figure 6. Total number of fish species versus watershed area for Ohio regional reference sites.
-------
salmonid species (4), headwater species (5), and sunfish and trout species
(8). These pool species decrease with increased degradation of pools and
instream cover (Gammon et al., 1981; Angermeier, 1983; Platts et al., 1983).
Most of these fishes feed on drifting and surface invertebrates and are
active swimmers. The sunfishes and salmonids are important sport species.
The sunfish metric works for most Mississippi Basin streams, but where sunfish
are absent or rare, other groups are used. Cyprinid species are used in
coolwater western streams; water column species occupy the same niche in
northeastern streams; salmonids are suitable in coldwater streams; headwater
species serve for midwestern headwater streams and trout and sunfish species
are used in southern Ontario streams. Karr et al. (1986) and Ohio EPA (1987b)
found the number of sunfish species to be dependent on stream size in small
streams, but Ohio EPA (1987b) found no relationship between stream size and
sunfish species in medium to large streams, nor between stream size and
headwater species in small streams. Scoring of this metric requires
development of species/waterbody size relationships.
8.8.2.4 Metric 4. Number and identity of sucker species (1). Substitutes:
Number of adult trout species (6), number of minnow species (5); and number of
sucker and catfish (8). These species are sensitive to physical and chemical
habitat degradation and commonly comprise most of the fish biomass in streams.
All but the minnows are long-lived species and provide a multiyear integration
of physical/chemical conditions. Suckers are common in medium and large
streams; minnows dominate small streams in the Mississippi Basin; and trout
occupy the same niche in coldwater streams. The richness of these species is
a function of stream size in small and medium sized streams, but not in large
rivers. Scoring of this metric requires development of species/waterbody size
relationships.
8.8.2.5 Metric 5. Number and identity of intolerant species (1).
Substitutes: Number and identity of sensitive species (5), amphibian species
(4); and presence of brook trout (8). This metric distinguishes high and
moderate quality sites using species that are intolerant of various chemical
and physical perturbations. Intolerant species are typically the first
species to disappear following disturbance. Species classified as intolerant
or sensitive should only represent the 5-10 percent most susceptible species,
otherwise this becomes a less discriminating metric. Candidate species are
determined by examining regional fishery books for species that were once
widespread but have become restricted to only the highest quality streams.
Ohio EPA (1987b) uses number of sensitive species (which includes highly
intolerant and moderately intolerant species) for head-water sites because
highly intolerant species are generally not expected in such habitats. Moyle
(1976) suggested using amphibians in northern California streams because of
their sensitivity to silvicultural impacts. This also may be a promising
metric in appalachian streams which may naturally support few fish species.
Steedman (1988) found that the presence of brook trout had the greatest
correlation with IBI score in Ontario streams. The number of sensitive and
intolerant species increases with stream size in small and medium sized
streams but is unaffected by size of large rivers. Scoring of this metric
requires development of species/waterbody size relationships.
160
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8.8.2.6 Metric 6. Proportion of tolerant individuals as green sunfish (1).
Substitutes: Proportion of individuals as common carp (2,4), white sucker
(3,4), tolerant species (5), creek chub (7), and dace (8). This metric is the
reverse of Metric 5. It distinguishes low from moderate quality waters.
These species show increased distribution or abundance despite the historical
degradation of surface waters, and they shift from incidental to dominant in
disturbed sites. Green sunfish are appropriate in small Midwestern streams;
creek chubs were suggested for central Appalachian streams; common carp were
suitable for a coolwater Oregon river; white sucker were selected in the
northeast and Colorado where green sunfish are rare to absent; and dace
(Rhinichthys species) were used in southern Ontario. To avoid weighing the
metric on a single species, Karr et al. (1986) and Ohio EPA (1987b) suggest
using a small number of highly tolerant species. Scoring of this metric may
require development of expectations based on waterbody size.
8.8.3 Trophic Composition Metrics
8.8.3.1 These three metrics assess the quality of the energy base and trophic
dynamics of the community. Traditional process studies, such as community
production and respiration, are time consuming to conduct and the results are
equivocal; distinctly different situations can yield similar results. The
trophic composition metrics offer a means to evaluate the shift toward more
generalized foraging that typically occurs with increased degradation of the
physicochemical habitat.
8.8.3.2 Metric 7. Proportion of individuals as omnivores (1,2,3,4,5,8).
Substitutes: Proportion of individuals as yearlings (4).
8.8.3.2.1 The percent of omnivores in the community increases as the physical
and chemical habitat deteriorates. Omnivores are defined as species that
consistently feed on substantial proportions of plant and animal material.
Ohio EPA (1987b) excludes sensitive filter feeding species such as paddlefish
and lamprey ammocoetes and opportunistic feeders like channel catfish. Where
omnivorous species are nonexistent, such as in trout streams, the proportion
of the community composed of yearlings, which initially feed omnivorously, may
be substituted.
8.8.3.3 Metric 8. Proportion of individuals which are insectivorous
cyprinids (1). Substitutes: Proportion of individuals as insectivore
(2,3,5), specialized insectivores (4), and insectivorous species (5);and
number of juvenile trout (4).
8.8.3.3.1 Insectivores or invertivores are the dominant trophic guild of most
North American surface waters. As the invertebrate food source decreases in
abundance and diversity due to physical/chemical habitat deterioration, there
is a shift from insectivorous to omnivorous fish species. Generalized
insectivores and opportunistic species, such as blacknose dace and creek chub
were excluded from this metric by Ohio EPA (1987b). This metric evaluates the
midrange of biotic integrity.
8.8.3.4 Metric 9. Proportion of individuals as top carnivores (1,3,8).
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Substitutes: Proportion of individuals as catchable salmonids (2), catchable
wild trout (4), and pioneering species (5).
8.8.3.4.1 The top carnivore metric discriminates between systems with high
and moderate integrity. Top carnivores are species that feed as adults
predominantly on fish, other vertebrates, or crayfish. Occasional piscivores,
such as creek chub and channel catfish, are not included. In trout streams,
where true piscivores are uncommon, the percent of large salmonids is
substituted for percent piscivores. These species often represent popular
sport fish such as bass, pike, walleye, and trout. Pioneering species are
used by Ohio EPA (1987b) in headwater streams typically lacking piscivores.
8.8.4 Fish Abundance and Condition Metrics
8.8.4.1 The last three metrics indirectly evaluate population recruitment,
mortality, condition, and abundance. Typically, these parameters vary
continuously and are time consuming to estimate accurately. Instead of such
direct estimates, the final results of the population parameters are
evaluated. Indirect estimation is less variable and much more rapidly
determined.
8.8.4.2 Metric 10. Number of individuals in sample (1,2,4,5,8).
Substitutes: Density of individuals (3,4).
8.8.4.2.1 This metric evaluates population abundance and varies with region
and stream size for small streams. It is expressed as catch per unit effort,
either by area, distance, or time sampled. Generally sites with lower
integrity support fewer individuals, but in some nutrient-poor regions,
enrichment increases the number of individuals. Steedman (1988) addressed
this situation by scoring catch per minute of sampling greater than 25 fish as
a three, and less than 4 fish as a one. Unusually low numbers generally
indicate toxicity, making this metric most useful at the low end of the
biological integrity scale. Hughes and Gammon (1987) suggest that in larger
streams, where sizes of fish may vary in orders of magnitude, total fish
biomass may be an appropriate substitute or additional metric.
8.8.4.3 Metric 11. Proportion of individuals as hybrids (1). Substitutes:
Proportion of individuals as introduced species (2,4), simple lithophils (5);
and number of simple lithophilic species (5).
8.8.4.3.1 This metric is an estimate of reproductive isolation or the
suitability of the habitat for reproduction. Generally as environmental
degradation increases, the percent of hybrids and introduced species also
increases, but the proportion of simple lithophils decreases. However, minnow
hybrids are found in some high quality streams, hybrids are often absent from
highly impacted sites, and hybridization is rare and difficult for many to
detect. Thus, Ohio EPA (1987b) substitutes simple lithophils for hybrids.
Simple lithophils spawn where their eggs can develop in the interstices of
sand, gravel, and cobble substrates without parental care. Hughes and Gammon
(1987) and Miller et al. (1988a) propose using percent introduced individuals.
This metric is a direct measure of the loss of species segregation between
162
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midwestern and western fishes that existed before the introduction of
midwestern species into western rivers.
8.8.4.4 Metric 12. Proportion of individuals with disease, tumors, fin
damage, and skeletal anomalies (1).
8.8.4.4.1 This metric depicts the health and condition of individual fish.
These conditions occur infrequently or are absent from minimally impacted
reference sites but occur frequently below point sources and in areas where
toxic chemicals are concentrated. They are excellent measures of the subacute
effects of chemical pollution and the aesthetic value of game and nongame
fish.
8.8.4.5 Metric 13. Total fish biomass (optional). Hughes and Gammon (1987)
suggest that in larger areas where sizes of fish may vary in orders of
magnitude this additional metric may be appropriate.
8.8.4.5.1 Because the IBI is an adaptable index, the choice of metrics and
scoring criteria is best developed on a regional basis through use of
available publications (Karr et al., 1986; Ohio EPA, 1987b; Miller et al.,
1988a). Several steps in the IBI process are common to all regions. The fish
species must be listed and assigned to trophic and tolerance guilds. Scoring
criteria are developed through use of high quality historical data and data
from minimally-impacted regional reference sites. The development of
reference sites have been accomplished for much of the country, but continued
refinements are expected as more fish community ecology data become available.
Once scoring criteria have been established, a fish sample is evaluated by
listing the species and their abundances (Figure 5), calculating values for
each metric and comparing these values with the scoring criteria. Individual
metric scores are added to calculate the total IBI score (Figure 7). Hughes
and Gammon (1987) and Miller et al. (1988a) suggest that scores lying at the
extremes of scoring criteria can be modified by a plus or minus; a combination
of three pluses or three minuses results in a two point increase or decrease
in IBI. Ohio EPA (1987b) scores proportional metrics as 1 when the number of
species and individuals in samples are fewer than 6 and 75, respectively, when
their expectations are of higher numbers.
8.9 Guidance for Use of Field Data Sheets
8.9.1 This subsection provides guidance for use of the bioassessment field
and laboratory data sheets. The guidance sheets give brief descriptions of
the information required for each data sheet.
8.9.2 Guidance for Header Information (Figure 8)
8.9.2.1 Water body Name: Name of stream or drain.
8.9.2.2 Location: Township, range, section, county where problem area is
located. For streams or drains; road crossings or outfall locations should be
referenced where applicable.
8.9.3 Reach/Milepoint: Indicate station reach/milepoint.
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8.9.4 Latitude/Longitude: Indicate station latitude/longitude.
8.9.5 County/State: Name of county and state where station is located.
8.9.6 Aquatic Ecoregion: Name of ecoregion.
8.9.7 Station: Agency name or number for station.
8.9.8 Investigators: List field personnel involved.
8.9.9 Date: Date of survey.
8.9.10 Agency: Agency name or affiliation (academic, private consulting)
8.9.11 Hydrologic Unit Code: Indicate the USGS cataloging unit number in
which the station is located.
8.9.12 Form Completed By: List personnel completing form.
8.9.13 Reason for Survey: The reason why this survey was conducted.
Station No.
Site
Scoring Criteria
Metrics(a)
1. Niuber of Native Fish Species
2. Nimber of Darter or Benthic Species
3. Nuflbet of Sunfish or Pool Species
4. HUB her of Sucker or Long-Lived Species
5. Nunber of Intolerant Species
6. X Green Sunfish or Tolerant Individuals
7. X Omnlvores
8. X Insectivores or Invertivores
9. X Top Carnivores
10. Total Nunber of Individuals
11. X Hybrids or Exotics
12. X Anonalies
Scorer
Couents:
5
i*y
>67
>67
>6?
>67
>67
<10
<20
>45
>5
>67
0
<1
3
1ST
33-67
33-67
33-67
33-67
33-67
10-25
20-45
20-45
1-5
33-67
0-1
1-5
1
(T) Metric Value Metric Score
<33
<33
<33
<33
<33
>25
>45
<20
<1
<33
>1
>5
IBI Score
(a) Rarr's original netrics or commonly used
ties.
(b) Karr's original scoring criteria or COMB
ecoregions.
substitutes.
only used substitutes.
See Figure 4 for other possibili-
These nay require refinenent in other
Figure 7. Data summary sheet for Fish Bioassessment II.
164
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en
Waterbody Name.
Reach/Milepoint _
County
State
Station Number.
Date
Time
Hydrologic Unit Code
Reason for Survey
Location.
Latitude/Longitude.
Aquatic Ecoregion
Investigators
Agency
Form Completed by.
Figure 8, Header information used for documentation and identification for sampling stations,
-------
8.10 Guidance for Impairment Assessment Sheet (Figure 4)
8.10.1 Detection of Impairment; Circle the one that applies.
8.10.2 Biological Impairment Indicator: Circle those that apply, as
indicated by the benthos, fish, and other aquatic biota.
8.10.3 Brief Description of Problem: Briefly explain the biological nature
of the problem, based on field investigation and sampling. List the year and
date of previous biological data and reports, and where the information can be
found (state file, BIOS).
8.10.4 Cause: Circle those that apply. Indicate which problem appears to be
the major cause of the stream impairment.
8.10.5 Estimated Areal Extent of Problem: Record estimated downstream extent
of impact (in m) and multiply by approximate stream width (in m) to estimate
area! width.
8.10.6 Suspected Source(s) of Problem: Check those that are suspected.
Briefly explain why you suspect a specific source, and reference other surveys
or studies done to document the problem and its source. Give title of
applicable report, author(s) and year published or completed. Use back of
sheet if necessary.
8.11 Guidance for Field Collection Data Sheet for Fish Bioassessment II
(Figure 5)
8.11.1 Drainage: Give name of stream or river and its basin site descriptor,
and unique site code.
8.11.2 Date: Enter day, month, and year of collection.
8.11.3 Sampling Duration: Record length of time in minutes actually
collecting fish. If replicates are taken, record them separately.
8.11.4 Sampling Distance: Measure, with a tape or calibrated range finder,
the length in meters of reach sampled.
8.11.5 Sampling Area: Multiply the length or reach sampled by the average
width sampled. Express in meters squared.
8.11.6 Crew: Indicate crew chief and crew members.
8.11.7 Habitat Complexity/Quality: Circle the descriptor that best describes
subjective evaluation of the physicochemical habitat.
8.11.8 Weather: Record air temperature, estimated wind velocity, percent
cloud cover, and precipitation.
8.11.9 Flow: Circle most appropriate descriptor.
166
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8.11.10 Information on Gear Used: Specify type, model, and number of
electrofisher, or the mesh size and length of seine, or concentration of fish
toxicant.
8.11.11 Gear/Crew Performance: Indicate effectiveness of crew in sampling
the site. Note problems with equipment, staff, or site obstacles, such as
extensive cover, high velocity current, excessive turbidity, floating debris,
deep muck or pools, or weather conditions. Electrofishing should be conducted
only during normal water flow and clarity conditions. Abnormally turbid
conditions are to be avoided as are elevated flow and current because these
conditions affect sampling efficiency. Also, if weather conditions are bad
(rain or high winds, lightning, etc.), electrofishing should be suspended
immediately or at the discretion of field personnel (Ohio EPA, 1990c).
8.11.12 Comments: Record any additional qualitative site data: sketch map
or take photographs, note the presence of springs, the evidence of fishing
activity, and/or potential or current impacts, the weather conditions (such as
evidence of recent high flows or unusually hot or cold weather immediately
preceding the survey), the biota observed (insect hatches, potential
vertebrate predators, the fish nesting and grazing sites, fish reproductive
conditions, or the fish seen but not captured.
8.11.13 Fish (preserved): Indicate if specimens were preserved for permanent
collection or further examination.
8.11.14 Number of Individuals; Number of Anomalies: Give total numbers of
fish and anomalies for the sample.
8.11.15 Genus/Species: Enter scientific name or unique standard abbreviation
for each species captured.
8.11.16 Adults (Number, Weight): Enter the number of adults of each species
and their total weight in grams. Individual or batch weight, depending on the
species' size and abundance. Species weight can also be determined by
weighing a subsample of individuals (20-30 fish spanning the size range
collected) and extrapolating for the total number of that species.
8.11.17 Juveniles (Number, Weight): Record the number of juveniles of each
species and their total weight as above. Juveniles and adults are
distinguished subjectively by coloration and size; the objective is to
determine whether both age classes are present,
8.11.18 Anomalies (Number): Indicate the number of fish by individual or
species, that are diseased, deformed, damaged, or heavily parasitized. These
are determined through careful external examination by a field-trained fish
biologist.
8.12 Guidance For Data Summary Sheet for Fish Bioassessment II (Figure 7)
8.12.1 Station Number: Indicate station number.
167
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8.12.2 Station Location: Record brief description of sampling site relative
to established landmarks (i.e., roads, bridges).
8.12.3 Metrics: List metrics used to conduct IBI calculations. Use either
Karr's original metrics or a published (or well supported) substitute
approach. Precede metric selection with analysis of reference site data or a
high quality historical database from a representative, large river basin.
8.12.4 Scoring Criteria: List published scoring criteria or use substitutes
where necessary. Analyze reference site data or historical data from a
representative large river basin before selecting criteria.
8.12.5 Metric Value: Record metric values (number or percent) for the
station. Metric values are obtained by comparing the collection data (Figure
5) with the tolerance and trophic guilds previously listed (Table 1). For
taxonomic metrics the numbers of different species are added, the total
number of individuals is recorded from the field collection data sheet.
Proportional metrics are determined by adding the number of individuals in
each category and dividing this total by the total number of individuals.
8.12.6 Metric Score: Score each metric by comparing the metric value for the
station with the previously chosen scoring criteria. Marginal values can be
given a plus or minus (see IBI score below).
8.12.7 Scorer: Enter the scorer's name.
8.12.8 IBI Score: The metric scores (and the pluses and minuses if used) are
added to give the IBI score. Three pluses or three minuses may increase or
decrease the IBI score by two points.
8.12.9 Comments: Metrics producing contrary results or suggestions for
improvement are entered here.
8.13 Habitat Assessment and Physical/Chemical Parameters
8.13.1 An evaluation of habitat quality is critical to any assessment of
ecological integrity. The habitat quality evaluation can be accomplished by
characterizing selected physical/chemical parameters and by systematic habitat
assessment. Through this approach, key parameters can be identified to
provide a consistent assessment of habitat quality. This evaluation of
habitat quality is relevant to all levels of rapid bioassessment.
8.13.2 Physical Characteristics and Hater Quality
8.13.2.1 Both physical characteristics and water quality parameters are
pertinent to characterization of the stream habitat. An example of the data
sheet used to characterize the physical characteristics and water quality of a
site is shown in Figure 9. The information requested includes measurements
made routinely during biological surveys. This phase of the survey is broken
into two sections: Physical Characterization and Water Quality (Figure 9).
These subsections are discussed separately below.
168
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cn
PHYSICAL CHABACTtBIIATIOB/WAIKB QUALITY
DMA SHEET
CHUUkCTKMIlATIOM
BIPAHIAB tOME/IHSTRCJUt fKATUBES
fradoainaat Surrounding Lund ila*:
For«»t Fi»I.d/3?*«fcttr« Agricultural fcaaidantial Co»«*rci»I
tocal Wat*r«h«d Eraxiom Boa a Mod*r*t* H**vy
Local M»fc*r«Ii»d SPS Foiiutioa: Bo *vid«nc* So** Potential 5ourc*s Obvious Soutc*s
B*tiait«d str*a* width n B«ti.nt*d Stcmim D«pth: KifCL* v tun n p
M*r*E rm-fT-^- * V«i0eity ^ DIM »r»«»nt: f»» ^ Ho
Industrial Oth*r
chaaneliied: le
Canopy Cov«c: Op«n Partly Opwa Partly Shaded Si
S E DIMKNT/S USSTEATE;
S*diA*nt odort: Bor»al 5*MA9« F»trol*ua Ch«»ic»l
S«di«»nt Oils; Aba.nt Slifbt «ad«tat» Ptofu*.
S*diM«nt D«poflits: aludg* s*vdu*t Pap*r Fib«t Sand R*liet sh*lla
Ara tb* undcrsidas o£ atouae which ara sot tlaaply a»ir»ddaJ56-s>« (JO in.)
6«-256-«» (2.5-10 in.)
2-6<~M [0.1-7.5 in.)
O.OS-J.OO-«. (gritty)
.0S4-.eS-M
<.004-e» (slick)
D*critu« stick*. Hood,
Coarse Plant
Materials (CVOH)
Ruck-Hud Black, V.ry fin*
orjsnic (rroH)
Marl amy, 3h«li
Fragwant*
P*rc*nt
Coapoai t ioa
in Stapling Ar«a
WATEB QUALITY
•T»iap»riituf*
Conductivity
at£i) used
Mater Odors ,* Rot*»l s*vt<$a I'
M«t«rSurfac«OJts: Slick Sh«*o
Turbidity: cl**c ail^btlyfyrbid
lwaB Ch*»ic*l
Globe f l*cks
furbid OgiBqu
VBATHIS COHDiriODS
r'HOTOGHAPH
OBSLRVATIOHS JMID/OI.
Figure 9, Physical characterization/water quality field data sheet for use with Fish Bioassessment II.
-------
8.13,2.2 Physical Characterization
8.13.2.2.1 Physical characterization parameters include estimations of
general land use and physical stream characteristics such as width, depth,
flow, and substrate. The evaluation begins with the riparian zone (stream bank
and drainage area) and proceeds instream to sediment/substrate descriptions.
Such information will provide insight as to what organisms may be present or
are expected to be present, and the presence of stream impacts. The
information requested in the Physical Characterization section of the Field
Data Sheet (Figure 9) is briefly discussed below.
8.13.2.2.2 Predominant Surrounding land Use; Observe the prevalent land-use
type in the vicinity (noting any other land uses in the area which, although
not predominant, may potentially affect water quality).
8.13.2.2.3 Local Watershed Erosion--The existing or potential detachment of
soil within the local watershed (the portion of the watershed that drains
directly into the stream) and its movement into a stream is noted. Erosion
can be rated through visual observation of the watershed and stream
characteristics. (Note any turbidity observed during water quality assessment
below.)
8.13.2.2.4 Local Watershed Nonpoint-Source Pollution—This item refers to
problems and potential problems other then siltation. Nonpoint source
pollution is defined as diffuse agricultural and urban runoff. Other
compromising factors in a watershed that may affect water quality or impacts
on the stream are feedlots, wetlands, septic systems, dams, and impoundments,
and/or mine seepage.
8.13.2.2.5 Estimated Stream Width (m): Estimate the distance from shore to
shore at a transect representative of the stream width in the area.
8.13.2.2.6 Estimated Stream Depth (m): riffle, run, and pool. Estimate the
vertical distance from water surface to stream bottom at a representative
depth at each of the three habitat types.
8.13.2.2.7 High Water Mark (m): Estimate the vertical distance from the
stream bank to the peak overflow level, as indicated by debris hanging in bank
or floodplain vegetation, and deposition of silt or soil. In instances where
bank overflow is rare, a high water mark may not be evident.
8.13.2.2.8 Velocity: Record an estimate of stream velocity in a
representative run area.
8.13.2.2.9 Dam Present: Indicate the presence or absence of a dam upstream
or downstream of the sampling station. If a dam is present, include specific
information relating to alteration of flow.
8.13.2.2.10 Channelized: Indicate whether or not the area around the
sampling station is channelized.
170
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8.13.2.2.11 Canopy Cover: Note the general proportion of open to shaded area
which best describes the amount of cover at the sampling station.
8.13.2.2.12 Sediment Odors: Disturb sediment and note any odors described
(or include any other odors not listed) which are associated with sediment in
the area of the sampling station.
8.13.2.2.13 Sediment Oils: Note the term which best describes the relative
amount of any sediment oils observed in the sampling area.
8.13.2.2.14 Sediment Deposits: Note those deposits described (or include any
other deposit not listed) which are present in the sampling area. Also
indicate whether or not the undersides of rocks which are not deeply embedded
are black in color (which generally indicates low dissolved oxygen or
anaerobic conditions).
8.13.2.2.15 Inorganic Substrate Components: Visually estimate the relative
proportion of each of the seven substrate particle types listed that are
present in the sampling area.
8.13.2.2.16 Organic Substrate Components: Indicate relative abundance of
each of the three substrate types listed.
8.13.2.3 Water Quality
8.13.2.3.1 Information requested in this Subsection (Figure 9) is standard to
many aquatic studies and allows for some comparison between sites.
Additionally, conditions that may significantly affect aquatic biota are
documented. It is important to document recent and current weather conditions
because of the potential impact that weather may have on water quality. To
complete this phase of the bioassessment, a photograph may be helpful in both
identifying station location and documenting habitat conditions. Any
observations or data not requested but deemed important by the field observer
should be recorded. This section is identical for all protocols and the
specific data requested are described below.
8.13.2.3.2 Temperature (°C), Dissolved Oxygen, pH, Conductivity: Measure and
record values for each of the water quality parameters indicated, using the
appropriate calibrated water quality instrument(s). Note the type of
instrument and unit number used.
8.13.2.3.3 Stream Type: Note the appropriate stream designation according to
State water quality standards.
8.13.2.3.4 Water Odors: Note those odors described (or include any other
odors not listed) that are associated with the water in the sampling area.
8.13.2.3.5 Water Surface Oils: Note the term that best describes the
relative amount of any oils present on the water surface.
8.13.2.3.6 Turbidity: Note the term which, based upon visual observation,
best describes the amount of material suspended in the water column.
171
-------
8.13.3 Habitat Quality and Assessment
8.13.3.1 The habitat assessment matrices (Figures 10 and 11) are taken from
Barbour and Stribling (1991). The habitat assessment matrix originally
published by Plafkin et al (1989) was based on the Stream Classification
Guidelines for Wisconsin developed by Ball (1982) and Methods of Evaluating
Stream, Riparian, and Biotic Conditions developed by Platts et al. (1983).
Also, see Subsection 8.16 for an example of a specific qualitative habitat
evaluation index field sheet (Figure 12) constructed for use by Ohio EPA.
Because this habitat assessment approach is intended to support biosurvey
analysis, the various habitat parameters are weighted to emphasize the most
biologically significant parameters. All parameters are evaluated for each
station studied. The ratings are then totaled and compared to a reference to
provide a final habitat ranking. Scores increase as habitat quality
increases. To ensure consistency in the evaluation procedure, descriptions of
the physical parameters and relative criteria are included in the rating form.
8.13.3.2 There is a great variability among streams; however, some
generalizations concerning similarities among stream types can be made
relative to gradient (Barbour and Stribling, 1991). Four generic stream
categories using gradient for establishing the framework can be identified:
montane, piedmont, valley/plains, and coastal plains. For these four
categories, two sets of parameters for assessing habitat quality have been
developed. For higher gradient streams there tends to be an increased
prevalence of riffles and runs. The matrix for "riffle/run prevalence" was
constructed (Barbour and Stribing, 1991) for use in montane and piedmont
streams (Figure 10). That for "glide/pool prevalence" (Figure 11) is for use
in valley/plains and coastal plains streams.
8.13.3.3 Reference conditions are used to normalize the assessment to the
"best attainable" situation. This approach is critical to the assessment
because stream characteristics will vary dramatically across different
regions. Other habitat assessment approaches may be used; or a more
rigorously quantitative approach to measuring the habitat parameters may be
used. However, the importance of a holistic habitat assessment to enhance the
interpretation of biological data cannot be overemphasized. A more detailed
discussion of the relationship between habitat quality and biological
condition is presented in Plafkin et al. (1989) and Barbour and Stribling
(1991).
8.13,3.4 Habitat parameters (Tabel 3) pertinent to the assessment of habitat
quality are separated into three principal categories: primary, secondary,
and tertiary. Primary parameters are those that characterize the stream
"microscale" habitat and have the greatest direct influence on the structure
of the indigenous communities. The primary parameters, which include
characterization of the bottom substrate and available cover, estimation of
embeddedness, estimation of the flow or velocity and depth regime, and canopy
cover have the widest score range (0-20) to reflect their contribution to
habitat quality. The secondary parameters measure the "macroscale" habitat
such as channel morphology characteristics. These parameters evaluate:
channel alteration, bottom scouring and deposition, and pool/riffle, run/bend
ratio, and lower bank channel capacity and have a range of 0-15. Tertiary
172
-------
Category
italUI PanmeWr
1, Bottom subatrat*
Instream cover
2 Embaddadness
3. «0. 15 cms (5 cist—
Plow at rep. tow
on
>0.l5cms
0.05 cm* (2 cfs)
Warn >0.15ems
(Sett)
16-20
Slow (<0.3 rrt's), d*ep
(>0,5 mj: slow, shallow
(<0,Sm!;last (>0.3
rrvi). deep; last, mallow
habitats all present.
ie-»
A mixture of conditions
where some areas ol
water surface fully
exposed lo sunlit, and'
other receiving various
degrees ol Uttered light.
16-20
Suo-Oi«i»m
JO- SON mix ol rubble,
gravel, or olher stable
habitat. Adequate
habitat.
11-15
Gravel, cobble, and
(Mulder panicles are
between 2S-50%
iurrounded by line
sediment.
11-15
0.03-0.05 cms
(1-2 ds)
0.05-0, 15 ems
(2-5 ets)
11-15
Only 3 ol the 4 habitat
categories present
(musing riffles or runs
receive tower score Wan
missing pools).
11-15
Covered by sparse
canopy; enlire water
surface receiving filtered
kgru.
11-15
MMtttMl
10-30*. mi» ol rubble,
gravel, or other stable
habitat. Habitat
availability tots than
desirable.
6-10
Gravel, cobble, and
boulder parades ire
between 50-75%
surrounded by tine
sediment.
6-10
0.01-0.03 cms
(.5-1 cfs)
0.03-0.05 om» (1-cfs)
6-10
Only 2 of the 4 habitat
categories present
(missing riffles or runt
receive lower score).
6-10
Compieieiy covered by
dense canopy; wa*er
surface completely
shaded OR nearly full
sunlight reaching waler
surface. Shading Smiled
to <3 hours per day.
6-10
Pan
Less than 10% rvtoOM.
gravel, or other KiMe
habitat. Lack ol riabtai
x obvious.
0-5
Gravel, cobble, and
Boulder particles are
over 75% surrounded
by fine sediment
0-5
<0.0i cms (.5 cfs)
<0 03 cms (1 cts)
0-5
Dominated by 1
velocity/depth category
{usually pools).
0-1
Lack of canopy, tuU
sunlight reaching water
surface.
0-5
S, Channflt alteration
ytu« « no enlargement Somd new increase m Mooeraie (Jeposilvon of Heavy deposits of fine
o( isiarxls or point bars, bar tormslion, mostly new gravel, coarse sand malarial, increased bar
and or no from coarse grave!; and/ on old and new bars; development; arxJ/or
or some channel^ al or, and or embankments on extensive
present, both banks. channelization.
12-15 6-11 4«? 0-3
Q _J
—K
UJ >>
UJ ID
a: of
t-O^x.
(/> UJ
CO
«s
«. Bottom scounng and Less than 5% ol trie 5-30% affected. Scour 30-50% affected. More than 50% of the
deposition bottom affected by at constrictions and Deposits and25. Essentially a
habilaL Repeat pattern pattern, Vwiery of or bend. Bottom straight stream.
of sequence relatively macrohatMat less than contours provide some Generally all flat water
frequent. optimal. habitat. or shallow riffle. Poor
habitat. ••
12-15 S-11 4-7 0-3
1 Overbank (lower) flows Cverbank (lower) flows Overbank (lower) flows Peak flows not
rare. Lowar bank W/t) occasional, W/D ratio common. W>D ratio contained or contained
ratio <7. (Channel width S-1S. 15-25. through channelization.
divided by depth or W/0 ratio >25.
height of tower bank.)
12-15 8-11 4-7 0-3
Upper bank stable. No
evidence of erosion or
bank failure, Side
slopes generally <30*.
Little potential for future
problems.
9-10
Over w% of me
streambank surfaces
covered by vegetation.
$-10
Vegetative disruption
minimal or not evident.
Almost all potential plant
bkxnass at present
stage of development
remains.
9-10
Dominant vegetation is
shrub.
9-10
> 1 8 meters.
S-10
Sow*
Moderately stable.
Infrequent, smaU areas
of erosion mostly healed
over. Side slopes up to
40* on one bank. Slight
potential in extreme
floods.
6-8
70- 89% of the
streambank surfaces
covered by vegetation.
6-8
Disruption evident but
not affecting community
vigor. Vegetative use is
moderate, and at least
one-half ol tne potential
plant biomass remains.
6-e
Dominant vegetation is
of tree form.
6-8
Between 12 and 16
meters.
6-8
Moderately unstable.
Moderate frequency and
size of eroaionaf areas.
Side slopes up lo 60*
on some banks. High
erosion potential during
extreme high flow,
3-S
SO- 79% at the
Ktreambank surfaces
covered by vegetation.
3-5
Disruption obvious;
some patches of bare
iot or dosety cropped
vegetation present. Less
fnan one-half of Ifce
potential plant biomass
remains.
3-5
Dominant vegetation is
grass or torbes.
3-S
Between 6 and 12
meters.
3-5
Unstable. Many eroded
areas, "flaw" areas
frequent along straight
sections and bends
Side slopes >60*
.common.
0-2
Less Dtan 50% of me
streambank surfaces
covered by vegetation.
0-2
Disruption ol
streambank vegetation
is very Ngn. Vegetation
has been removed to 2
inches or less in
average stubble height
0-2
Over 50% of me
itreambank has no
vegetation and
dominant material it
soil, rock, bridge
materials, culverts, or
mine tailings.
0-2
<6 meters.
0-2
'
(V
o
c
OJ
s-
o.
c=
3
S-
QJ
QJ
OJ
I/)
O) "O
1/1
0)
O)
s-
CJ>
173
-------
CO
i—i
25.
0-3
Unstable. Many eroded
areas. "Flaw" areas
frequent along straight
sections and bends.
Side slopes >60°
common.
0-2
Less than 50% of the
streambank surfaces
covered by vegetation.
0-2
Disruption of
streambank vegetation
Almost ail potential plant vigor. Vegetative use is
biomass at present moderate, and at least
stage of development one-half of the potential
remains. plant biomass remains.
9-10
soil or closely cropped is very high. Vegetation
vegetation present. Less has been removed to 2
than one-half of the inches or less in
potential plant biomass average stubble height.
remains.
6-8
3-5
0-2
11. Streamside cover
Dominant vegetation is
shrub.
Dominant vegetation is
of tree form.
Dominant vegetation is
grass or forbes.
9-10
6-8
3-5
Over 50% of the
streambank has no
vegetation and.
dominant material is
soil, rock, bridge
materials, culverts, or
mine tailings.
0-2
cu
o
C
cu
(9
>
CD
s_
o.
o
o
Q.
Ol
OJ -
CD— ^
-Ci-H
CTi
fBi— i
I i *^^
fO
"O CD
a>_a
41 si
4->
•*->(/}
C
cu-o
£ C
(/) ft)
tfl
a> s-
3
nt
nj OQ
.a
as
CD
S-
3
12. Riparian vegetative
zone width (least
buffered side)
>18 meters.
Between 12 and 18
meters.
Between 6 and 12
meters.
9-10
<6 meters.
6-8
3-5
0-2
Column Totals
Score.
174
-------
QHEi Score:
RM
Stream
Location
1] SUBSTRATE (Chock ONLYlwo Substrate TYPE BOXES; Estimate % or note ivtry type presanl);
Dale
RivwCode
Scorers Name:
TYPE POOL RIFFLE . POOL RIFFLE SUBSTRATE ORlGIl
:P O-BLDR /SLABSI1 0] O O-GRAVEL [7] Check ONE (OK 2 i AVE
PO-80ULDER(9]
O P-COBBLE [8j
O P-HARDPAN [4]
OP-MUCK [2]
0-0-SILT|2]
O O-SAND [6] - :'-,:
00-BEDROCKIS]
PP-DETRrrUS[3J
O O-ARTIFiCIAL[OJ
NOTE: (Ignore sludge that originates from point-sources;
score on natural substrates) -;-'O-S or More {2}
NUMBER OF SUBSTRATE TYPES: :;O-4 or Less [0]
COMMENTS
O -LIMESTONE [1]
0 -TILLS [11
P:-WETLANDSf0)
O-HARDPANIQ]
0. -SANDSTONE [OJ
P :-RtP/RAP [0]
piLACUSTRINEp]
6 -SHALE [-11
P-COAL FINES f-21
!i , SUBSTRATE QUALITY
flAGE) Check ONE (OR 2 S AVERAGE)
SILT: O -SILT HEAVY (-2]
O -SILT MODERATE [-11
O -SILT NORMAL pi
__ J2:§1LT£REE{1]
EMBEDDED O -EXTENSIVE [-2]
NESS: 0 -MODERATE HI
O -NORMAL [0)
0 -NONE [1J
Substrate
Mas.
2] WSTREAM COVER
TYPE: (Check All That Apply)
j3 :-UNDERCUT BANKS [1J P -DEE? POOLS> 70 cm [2p -OXBOWS [11
"0 OVERHANGING VEGETATION [1] O-ROOTWADS [1] O-AQUATIC MACROPHYTES [1]
P -SHALLOWS (IN SLOW WATER) [1] O -BOULDERS [1] O -LOGS OR WOODY DEBRIS [1J
;P-RQOTMATS(1] COMMENTS:
AMOUNT: (Check OM-KOne or
check 2 and AVERAGE}
O-EXTENSIVE > 75% [11]
O - MODERATE 25-75% [7j
O-SPARSE5-25% [3]
O - NEARLY ABSENT < 5%(1J
Cover
3] CHANNEL MORPHOLOGY: (Check OWL XQne PER Category OR check 2 and 4V£r?4G£)
DEVELOPMENT
O-EXCELLENT [7[
O-GOOD [5] '
P • FAIR [3]
• P-POOR (11V
SINUgSiTY
:0 -HIGH HI
:.&-MOOEHATE[3|
.O-LOW[2l
b;-NONE [11
COMMENTS:
41 RIPARIAN ZONE AND BANK EROSION
giPARIANWlDTH
L R (Per Bank)
OO'-W!DE>5Qm[4]
O O - MODERATE 10-50m [3]
OO"-NARROW5-10m[2l
Op"- VERY NARROW 1m!Si ';••:•
P- 0.7-1 m [4]
P • Q.4-0,7m [2]
P - 0.2-0.4m [1]
P • < 02m [POOL=0]
MORPHOLOGY
(Check! or 2 4 AVERAGE)
P'-POOL WIDTH > RIFFLE WIDTH [2J
P -POOL WIDTH . RIFFLE WIDTH [1]
P'-POOL WIDTH < RIFFLE W. [OJ
COMMENTS:
CURRENT VELOCITY TOOL & RIFFLES!!
(Check 4tf That Apply)
O"-EDDIES[1] CT-TORRENTIAL[-1]
P--FAST[1J P'-INTEHSTITIALH]
O"-MODERATE [1 ]. P"-INTE?.MITTENT(-2]
P'-SLOW[1]
Max 12
RIFFLE/RUN PEPTH
P - GENERALLY >10 cm.MAX > 50 (4|
P - GENERALLY >10 cm; MAX < 50[3j
P- GENERALLY 5-10 cm{1J
O - GENERALLY < 5 cm [RIFaE=OJ
COMMENTS:
• CHECK ONE OR CHECK 2 AND AVERAGE
RIFFLE/RUN SUBSTRATE
P-STABLE (e.g., Cobble, Boulder) P]
P-MOD. STABLE (e.g.,Large Gravel) [1j
D-UNSTABLE (Fine Gravel.Sand) [0]
6] GRADIENT (ft/mi):.
DRAINAGE AREA (sq.mi.):_
%POOL
%RIFFLE:
P - NONE [2]
P-LOW[1J
P-MODERATE [0]
P - EXTENSIVE [-I]
O- NO RIFFLE [Metrical
%GLIOE;
%RUN: I I
Riffle/Run
Figure 12. Example of Ohio EPA (1991) qualitative habitat evaluation index
field sheet. 175
Max 3
Gradient
Max 10
-------
is Sampling Reach Representative of Stream? (Y/N) If Not, Exp!ain:_
Additional Comr,.onis/Pollution Impacts;
First Sampling Pass:
Second Sampling Pass:
Third-Sampling Pass:
CANOPY (% OPEN)
Gear
Distance
Water Clarity
Water Stage
GRADIENT: O-LOW O-MODERATE O-HIGH
Subjective Rating Aesthetic Sating
(1-10) (1-10)
PHOTOS:
STREAM MEASUREMENTS:
AVERAGE WIDTH:
AVERAGE DEPTH:
MAXIMUM DEPTH:
DRAWING OF STREAM:
FLOWC
Figure 12. Example of Ohio EPA (1991) qualitative habitat evaluation index
field sheet (continued).
176
-------
parameters evaluate riparian and bank structure and comprise four parameters:
upper bank stability, bank vegetative stability, streamside cover, and width
of riparian vegetative zone. These tertiary parameters are most often ignored
in biosurveys. The tertiary parameters have a score range of 0-10.
8.13.3.5 Habitat evaluations (Table 3) are first made on instream habitat,
followed by channel morphology, and finally on structural features of the bank
and riparian vegetation. Stream segment length or area assessed will vary
with each site. Generally, primary parameters are evaluated within the first
riffle/pool sequence, or the immediate sampling area such as in the case of
fish sampling. Secondary and tertiary parameters are evaluated over a larger
stream area, primarily in an upstream direction where conditions will have the
greater impact on the community being studied. The actual habitat assessment
process involves rating each of the nine parameters as either: excellent,
good, fair, or poor based on the criteria included on the Habitat Assessment
Field Data Sheet (Figures 10 and 11).
8.13.3.6 A total habitat score is obtained for each biological station and
compared to a site-specific control or regional reference station. The ratio
between the score for the station of interest and the score for the control or
regional reference provides a percent comparability measure for each station
Table 3). The station is then classified on the basis of its similarity to
expected conditions (as represented by the control or reference station), and
its inferred potential to support an acceptable level of biological comminity
health.
8.13.3.7 The use of a percent comparability evaluation (Table 3) allows for
regional and stream-size differences which affect flow or velocity, substrate,
and channel morphology. Some regions are characterized by streams having a
lower channel gradient. Such streams are typically shallower, have a greater
pool/riffle or run/bend ratio, and less stable substrate than streams with a
steep channel gradient. Although some low gradient streams do not provide the
diversity of habitat or fauna afforded by steeper gradient streams, they are
characteristic of certain regions. Use of the matrix presented as Figure 14
can allow more direct evaluation of low gradient streams relative to regional
expectations.
8.13.3.8 Listed below is a general explanation for each of the twelve habitat
parameters to be evaluated for riffle/run prevalent streams (higher gradient,
Figure 10).
8.13.3.9 Primary Parameters-Substrate and Instream Cover
8.13.3.9.1 The primary instream habitat characteristics directly pertinent to
the support of aquatic communities consist of substrate type and stability,
availability of refugia, and migration/passage potential. These primary
habitat parameters are weighted with the highest weighting reflective of their
degree of importance to the biological communities.
1. Bottom Substrate/Instream Cover--This refers to the availability of
habitat for support ofaquatic organisms. A variety of substrate materials
and habitat types is desirable. The presence of rock and gravel in flowing
177
-------
TABLE 3. NINE HABITAT PARAMETERS AND ASSESSMENT CATEGORY
Condition/Parameter
PRIMARY-SUBSTRATE AND INSTREAM COVER
1. Bottom substrate/instream cover
2. Embeddedness
3. Flow/velocity/depth
4. Canopy cover (shading)
SECONDARY-CHANNEL MORPHOLOGY
5. Channel alteration
6. Bottom scouring and deposition
7. Pool/riffle, run/bend ratio
8. Lower bank channel capacity
TERTIARY-RIPARIAN AND BANK STRUCTURE
9. Upper Bank stability
10. Bank vegetative stability (grazing/
disruptive pressure)
11. Streamside cover
12. Riparian vegetative zone width
Assessment Category
Comparable to Reference
Supporting
Partially Supporting
Non-Supporting
Excellent
16-20
16-20
16-20
16-20
12-15
12-15
12-15
12-15
9-10
9-10
9-10
9-10
Condi
Good
11-15
11-15
11-15
11-15
8-11
8-11
8-11
8-11
6-8
6-8
6-8
6-8
tion
Fair
6-10
6-10
6-10
6-10
4-7
4-7
4-7
4-7
3-5
3-5
3-5
Poor
0-5
0-5
0-5
0-5
0-3
0-3
0-3
0-3
0-2
0-2
0-2
3-5 0-2
Percent of
Comparability
>90%
75-89%
60-74%
<59%
178
-------
streams is generally considered the most desirable habitat. However,
other forms of habitat may provide the niches required for community
support. For example, logs, tree roots, submerged or emergent vegetation,
undercut banks, etc., will provide excellent habitat for a variety of
organisms, particularly fish. Bottom substrate is evaluated and rated by
observation.
2. Embeddedness--The degree to which boulders, rubble, or gravel are
surrounded by fine sediment indicates suitability of the stream substrate
as habitat for benthic macroinvertebrates and for fish spawning and egg
incubation. Embeddedness is evaluated by visual observation of the degree
to which larger particles are surrounded by sediment. In some western
areas of the United States, embeddedness is regarded as the stability of
cobble substrate by measuring the depth of burial of large particles
(cobble, boulders).
3. Stream Flow and/or Stream Velocity--Stream flow relates to the ability of
a stream to provide and maintain a stable aquatic environment. Stream
flow (water quantity and gradient) is most critical to the support of
aquatic communities when the representative low flow is <0.15 cms (5 cfs).
In these small streams, flow should be estimated in a straight stretch of
run area where banks are parallel and bottom contour is relatively flat.
Even where a few stations may have flows in excess of 0.15 cms, flow may
still be the predominate constraint. Therefore, the evaluation is based
on flow rather than velocity.
4. Canopy Cover (Shading)--Shading, as provided by canopy cover, is important
for the control of water temperature, its effect on biological processes
in general, and as a factor in photosynthetic activity and primary
production. A diversity of shade conditions is considered optimal, that
is, with some areas of the sampling station receiving direct sunlight,
others, complete shade, and other, filtered light.
8.13.3.10 In larger streams and rivers (> 0. 15 cms), velocity, in
conjunction with depth, has a more direct influence than flow on the structure
of benthic communities (Osborne and Hendricks, 1983) and fish communities
(Oswood and Barber, 1982). The quality of the aquatic habitat can, therefore,
be evaluated in terms of a velocity, and depth relationship. As patterned
after Oswood and Barber (1982), four general categories of velocity and depth
are optimal for benthic and fish communities: (1) slow (<0.3 m/s), shallow
(<0.5 m); (2) slow (<0.3 m/s), deep (>0.5 m); (3) fast (>0.3 m/s), deep (>0.5
m); and (4) fast (>0.3 m/s), shallow (<0.5 m). Habitat quality is reduced in
the absence of one or more of these four categories.
8.13.3.11 Secondary Parameters-Channel Morphology
8.13.3.11.1 Channel morphology is determined by the flow regime of the
stream, local geology, land surface form, soil, and human activities (Platts
et al. 1983). The sediment movement along the channel, as influenced by the
tractive forces of flowing water and the sinuosity of the channel, also
affects habitat conditions.
179
-------
S. Channel Alteration—The character of sediment deposit from upstream is an
indication of the severity of watershed and bank erosion and stability of
the stream system. The growth or appearance of sediment bars tends to
increase in depth and length with continued watershed disturbance.
Channel alteration also results in deposition, which may occur on the
inside of bends, below channel constrictions, and where stream gradient
flattens out. Channelization (e.g., straightening, construction of
concrete embankments) decreases stream sinuosity, thereby increasing
stream velocity and the potential for scouring.
6. Bottom Scouring and Deposition—These parameters relate to the destruction
of instream habitat resulting from the problems described above.
Characteristics to observe are scoured substrate and degree of siltation
in pools and riffles. Scouring result from high velocity flows. The
potential for scouring is increased by channelization. Deposition and
scouring result from the transport of sediment or other particulates and
may be an indication of large scale watershed erosion. Deposition and
scouring is rated by estimating the percentage of an evaluated reach that
is scoured or silted (i.e., 50-ft silted in a 100-ft stream length equals
50 percent).
7. Pool/Riffle, Run/Bend Ratio—These parameters assume that a stream with
riffles or bends provides more diverse habitat than a straight (run) or
uniform depth stream. Bends are included because low gradient streams may
not have riffle areas, but excellent habitat can be provided by the
cutting action of water at bends. The ratio is calculated by dividing the
average distance between riffles or bends by the average stream width. If
a stream contains riffles and bends, the dominant feature with the best
habitat should be used.
8. Lower bank channel capacity--This parameter is designed to allow
evaluation of the ability of a stream channel to contain normal peak
flows, since the lower bank is that over which water initially escapes,
it is the focus of this individual parameter.
8.13.3.12 Tertiary Parameters-Riparian and Bank Structure
8.13.3.12.1 Well-vegetated banks are usually stable regardless of bank
undercutting; undercutting actually provides excellent cover for fish (Platts
et al., 1983). The ability of vegetation and other materials on the
streambanks to prevent or inhibit erosion is an important determinant of the
stability of the stream channel and instream habitat for indigenous organisms.
Because riparian and bank structure indirectly affect the instream habitat
features, they are weighted less than the primary or secondary parameters.
8.13.3.12.2 Tertiary parameters are evaluated by observation of both upper
and lower bank characteristics. The upper bank is the land area from the
break in the general slope of the surrounding land to the normal high water
line. The upper bank is normally vegetated and covered by water only during
extreme high water conditions. Land forms vary from wide, flat floodplains to
narrow, steep slopes. The lower bank is the intermittently submerged portion
180
-------
of the stream cross section from the normal high water line to the lower water
line. The lower channel defines the stream width.
9. Upper Bank Stability—Bank stability is rated by observing existing or
potential detachment of soil from the upper and lower stream bank and its
potential movement into the stream. Steeper banks are generally more
susceptible to erosion and failure, and may not support stable
vegetation. Streams with poor banks will often have poor instream
habitat. Adjustments should be made in areas with clay banks where
steep, bare areas may not be as susceptible to erosion as other soil
types.
10. Bank Vegetative Stability (Grazing/Disruptive Pressure)--Vegetative
stability is evaluated here as it relates to reduction of erosion and
biological contribution to the aquatic ecosystem. Bank soil is generally
held in place by plant root systems. Erosional protection may also be
provided by boulder, cobble, or gravel material. Areas of higher
vegetative coverage receive higher ratings (Ball, 1982; Platts et al.,
1983). An estimate of the density of bank vegetation (or proportion of
boulder, cobble, or gravel material) covering the bank provides an
indication of bank stability and potential instream sedimentation.
Vegetative stability is best rated in areas of little riparian zone
disturbance. Areas exposed to grazing pressures or other disruption
should be evaluated under the second set of conditions. Grazing or other
disruptive pressure is evaluated in terms of the potential plant biomass
at the site in any given season.
11. Streamside Cover--Streamside cover vegetation is evaluated in terms of
provision of stream-shading; and escape cover or refuge for fish. A
rating is obtained by visually determining the dominant vegetation type
covering the exposed stream bottom, bank, and top of bank. Platts (1974)
found that Streamside cover consisting primarily of shrub had a higher
fish standing crop than similar-size streams having tree or grass
Streamside cover. Riparian vegetation dominated by shrubs and trees
provides the course particulate organic matter (CPOM) source in
allochthonous systems.
12. Riparian Vegetative Zone Width (Least Buffered Side)--The riparian buffer
zone is rated by its width on the side with the nearest disturbance or
human influence. Increasing buffer zone width is positively correlated
with shade. Vegetated buffer zones are also effective in removal of
particulate pollutants from storm runoff, can reduce runoff velocity and
volume, and can aid in the recharging of groundwater.
8.13,3.12 The matrix constructed for lower gradient streams likely to be
encountered is coastal plains and prairie regions (Figure 11; Barbour and
Stribing, 1991) differs from Figure 10 by two parameters. The following two
parameters (numbers 2 and 3) have been added to emphasize the increased
importance of pools as habitat in these streams,
2. Pool Substrate Characterization—diversity and variability in substrate
181
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particle size are rated higher than uniform particle sizes in pool
substrates.
3. Pool Variability—This parameter rates the mixture of pool sizes within a
stream reach. Variability in pool sizes will support a healthy fisheries
and a more diverse benthic macroinvertebrate assemblage.
8.13.3.13 Additional Habitat Assessment Considerations
8.13.3.13.1 Two additional variables are important and should be considered
by the investigator: (1) seasonal aspects of habitat evaluation; and (2) the
length of the stream reach to be evaluated for habitat quality. To properly
address both of these considerations, the major objective of the habitat
assessment should be identified. If the habitat assessment is being conducted
in relation to the biological collections, all field assessments and
collections should be performed concurrently, and the sampling domain (site
boundaries) should be critically established. On the other hand, if the
purpose of the habitat assessment is to characterize or classify a stream or
watershed, a different sampling regime or criterion might be established.
8.13.13.2 With regard to seasonality, it is important to understand that the
habitat quality may change depending on the time of the assessment. However,
the primary habitat parameters amy change most dramatically, having the
greatest influence on the communities under study. This particular habitat
assessment approach is designed as a tool for evaluating the potential
biological condition of the communities. With this in mind, the actual
sampling site where the resident communities are being collected is of central
importance in the habitat evaluation. The sampling site should be evaluated
for the primary habitat parameters.
8.13.13.3 The stream reach upstream of the site should be included in the
evaluation of the secondary and tertiary parameters. The actual delineation
of the length of the reach will depend on the objectives of the study. For
nonpoint source assessment, the reach may be much as a half mile; for point
source evaluations, the reach may be only a few hundred yards. In the
assessment of the fish community, a downstream reach amy be incorporated onto
the habitat evaluation for the primary and secondary parameters.
8.14 Selected References for Determining Fish Tolerance, Trophic,
Reproductive, and Origin Classifications (Also, See Section 12,
Fisheries Bibliography)
ALABAMA
Smith-Vaniz, W.F. 1987. Freshwater fishes of Alabama. Auburn University
Agricultural Experiment Station, Auburn, AL. 209 pp.
ALASKA
McPhail, J.D. and C.C. Lindsey. 1970. Freshwater fishes of northestern
Canada and Alaska. Bulletin No. 173. Fisheries Researd Board of
Canada. 381 pp.
182
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Morrow, J.E. 1980. The freshwater fishes of Alaska. Alaska Northwest
Publishing Company, Anchorage, AK. 300 pp.
ARIZONA
Minckley, W.L. 1973. Fishes of Arizona., Arizona Game and Fish
Department, Phoenix, AZ. 293 pp.
ARKANSAS
Black, J.D. 1940. The fishes of Arkansas. Ph.D. Thesis, Univ. of
Michigian Microfilm, Ann Arbor, MI.
Buchanan, T.M. 1973. Key to the fishes of Arkansas. Arkansas Game and
fish Commission, Little Rock, AK. 68 pp., 198 maps.
Robison, H.W. and T.M. Buchanan. 1988. The fishes of Arkansas. Univ.
Arkansas Press, Fayetteville, AK.
CALIFORNIA
Moyle, P.B. 1976. Inland fishes of California. University of California
Press, Berkeley, CA. 405 pp.
COLORADO
Beckman, W.C. 1953. Guide to the fishes of Colorado. Leaflet No. 11.,
University of Colorado Museum. 110 pp.
Everhart, W.H. and W.R. Seaman. 1971. Fishes of Colorado. Colorado Game,
Fish, and Parks Division, Denver, CO. 77 pp.
CONNECTICUT
Whitworth, W.R., P.L. Berrien, and W.T. Keller. 1968. Freshwater fishes of
Connecticut. Bulletin No. 101. State Geological and Natural History
Survey of Connecticut. 134 pp.
DELAWARE
Lee, D.S., S.P. Platania, C.R. Gilbert, R. Franz, and A. Norden. 1981. A
revised list of the freshwater fishes of Maryland and Delaware.
Proceedings of the Southeastern Fishes Council 3:1-10.
FLORIDA
Briggs, J.C. 1958. A list of Florida fishes and their distribution. Bulletin
of the Florida State Museum 1(8):223-318.
Gilbert, C.P., G.H. Burgess, and R.W. Yerger. In preparation. The freshwater
fishes of Florida.
183
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GEORGIA
Dahlberg, M.D., and D.C. Scott. 1971. The freshwater fishes of Georgia.
Bulletin of the Georgia Academy of Science. 19:1-64.
IDAHO
Simpson, J.C. and R.L. Wallace. 1982. Fishes of Idaho. The University of
Idaho Press, Moscow, ID. 238 pp.
ILLINOIS
Forbes, S.A. and R.E. Richardson. 1908. The fishes of Illinois. Illinois
State Laboratory of Natural History. 357 pp., plus separate atlas
containing 102 maps.
Forbes, S.A. and R.E. Richardson. 1920. The fishes of Illinois. Second
edition. Illinois Naural History Survey. 357 pp.
Smith, P.M. 1979. The fishes of Illinois. Illinois State Natural History
Survey, University of Illinois Press, Urbana, IL. 314 pp.
INDIANA
Gerking, S.D. 1945. The distrbution of the fishes of Indiana. Investigation
lakes and streams 3:1-137.
Simon, T.P., J.O. Whitaker, J. Castrale, and S.A. Minton. 1992. Checklist of
the vertebrates of Indiana. Proc. Ind. Acad. Sci. In Press.
IOWA
Bailey, R.M. 1956. A a revised list of the fishes of Iowa with keys for
identification. Iowa State Conservation Commission, Des Moines, IA
Harlan, J.R. and E.B. Speaker. 1951. Iowa fish and fishing. State
Conservation Commission, State of Iowa. 237 pp.
KANSAS
Cross, F.B. 1967. Handbook of fishes of Kansas. Public Education Series No.
3. University of Kansas Museum of Natural History 189 pp.
KENTUCKY
Burr, B.M. 1980. A distribution checklist of the fishes of Kentucky.
Brimeyana 3:53-84.
Burr, B.M. 1986. A distributional atlas of the fishes of Kentucky. Kentucky
Nature Preserves Commission Sci. and Tech. Series No. 4. 398 pp.
184
-------
Clay, W.M 1975. The fishes of Kentucky. Kentucky Department of Fish and
Wildlife Resources, Frankford, KY. 416 pp.
LOUISIANA
Douglas, N.H. 1974. Freshwater fishes of Louisiana. Claitors Publishing
Division, Baton Rouge, LA. 443 pp.
MAINE
Everhart, W.H. 1966. Fishes of Maine. Third edition. Maine Department of
Inland Fisheries and Game, Augusta, ME, 96 pp.
MARYLAND
Elser, H.J. 1950. The common fishes of Maryland. Chesapeake Biological
Laboratory, Solomons Island, MD.
Lee, D.S., S.P, Platania, C.R. Gilbert, R. Franz, and A. Norden, 1981. A
revised list of the freshwater fishes of Maryland and Delaware.
Proceedings of the Southeastern Fishes Council 3:1-10.
MASSACHUSETTS
Mugford, P.S. 1969. Illustrated manual of Massachusetts freshwater fish.
Massachusetts Division of fish and Game, Boston, MA. 127 pp.
MICHIGAN
Hubbs, C.L. and G.P. Cooper. 1936, Minnows of Michigan. Bulletin of
Cranbrook Institute Science 8:1-99.
Hubbs, C.L. and K.F. Lagler. 1946. Fishes of the Great Lakes region.
Cranbrook Institute of Science, Bloomfield Hills, Mi. 186 pp.
Taylor, W.R. 1954. Records of fishes in the John N. Lowe collection from the
Upper Penninsula of Michigan. Miscellaneous Publications of the
Museum of Zoology, University of Michigan 87:5-49
MINNESOTA
Eddy. S. and J.C. Underhill. 1974, Northern Fishes, with special reference
to the Upper Missippi Valley. University of Minnesota Press,
Minneapolis, Minnesota. 414 pp.
Philips, G.L. and J.C. Underhill. 1971, Distribution and variation of the
Catostomidae of Minnesota. Occasional Papers of the Bell Museum of
Natural History 10:1-45.
Underhill, J.C. 1957, The distribution of Minnesota minnows and darters in
relation to Pleistocene glaciation. Occasional Papers of the Minnesota
Museum of Natural History 7:1-45.
185
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MISSISSIPPI
Clemmer, G.H., R.D. Suttkus, and J.S. Ramsey, 1975. A preliminary checklist
of endangered and rare fishes of Mississippi, in preliminary list of
rare and threatened vertebrates in Mississippi. Mississippi Game and
Fish Commission, pp. 6-22.
Cook, F.A. 1959. Freshwater fishes in Mississippi. Mississippi Game and
Fish Commission, Jackson, MS. 239 pp.
MISSOURI
Pflieger, W.L. 1971. A distribution study of Missouri fishes. University of
Kansas Museum of Natural History, Publication 20(3):225-570.
Pflieger, W.L. 1975. The fishes of Missouri. Missouri Department of
Conservation, Columbia, MO. 343 pp.
MONTANA
Brown, C.J. D. 1971. Fishes of Montana. Montana State University,
Bozeman, Montana. 207 pp.
NEBRASKA
Johnson, R.E. 1941. The distribution of Nebraska fishes. Ph.D. dissertation.
University of Michigan Library.
Morris. J.L. and L. Witt. 1972. the fishes of Nebraska. Nebraska Game and
Parks Commission, Lincoln, NB. 98 pp.
NEVADA
LaRivers, I. 1962. Fish and fisheries of Nevada. Nevada State Fish and Same
Commission, Carson City, NV. 782 pp.
NEW HAMPSHIRE
Scarola, J.F. 1973. Freshwater fishes of New Hampshire. New Hampshire
Fish and Game Department, Concord, NH. 131 pp.
NEW JERSEY
Stiles, E.W. 1978. Vertebrates of New Jersey. Edmund W. Stiles Publishers,
Somerset, NJ. 148 pp.
NEW MEXICO
Koster, W.J. 1957. Guide to the fishes of New Mexico. University of New
Mexico Press, Albuquerque, NM. 116 pp.
186
-------
Sublette, J.E., M.D. Hatch, and M. Sublette. 1990. The fishes of New Mexico.
Univ. New Mexico Press, Albuquerque, NM. 393 pp.
NEW YORK
Decker, D.J., R.A. Howare, Jr., W.E. Everhart, and J.W. Kelley. 1982.
Guide to freshwater fishes of New York. Cornell University,
Distribution Center, Ithaca, NY.
Greeley, J.R. 1927-1940. Watershed survey reports on fishes of New York
rivers, published as supplements to the 16th through 29th Annual Reports
of the New York State Conservation Department, Albany, NY.
Smith, C.L. 1985. Inland fishes of New York. New York State Dept. Environ.
Conservation, Albany, NY. 522 pp.
NORTH CAROLINA
Menhinick, E.F., T.M. Burton, and J.R. Bailey. 1974. An annotated checklist
of the freshwater fishes of North Carolina. Journal of the Elisha
Mitchell Scientific Society 90(1):24-50.
Menhinick, E.F. 1991. The freshwater fishes of North Carolina. Univ. North
Carolina, Charlotte, NC.
NORTH DAKOTA
Hankinson, T.L. 1929. Fishes of North Dakota. Papers of the Michigan
Academy of Science, Arts, and Letters 10:439-460.
OHIO
Trautman, M.B. 1981. The fishes of Ohio. Ohio State University Press,
Columbus, OH. 683 pp.
Ohio EPA. 1978. Appendix B: Development of fish community IBI metrics. In:
Biological criteria for the protection of aquatic life: Volume II:
Users manual for biological field assessment of Ohio surface waters.
Ohio EPA, Division Water Quality Monitoring and Assessment, 1800
watermark Drive, P.O. Box 1049, Columbus, OH.
OKLAHOMA
Miller, R.J. and H.W. Robinson. 1973. The fishes of Oklahoma. Oklahoma
State University Press, Stillwater, OK. 246 pp.
OREGON
Bond, C.E. 1973. Keys to Oregon freshwater fishes. Technical Bulletin 58:1-
42. Oregon State University Agricultural Experimental Station,
Corvallis, OR.
187
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PENNSYLVANIA
Cooper, E. L. 1983. Fishes of Pennsylvania and the northeastern United
States. Pennsylvania State Press, University Park, PA. 243 pp.
Fowler, H.W. 1940. A list of the fishes recorded from Pennsylvania.
Bulletin of the Pennsylvania Board of Fish Commission 7:1-25.
SOUTH CAROLINA
Anderson, W.D. 1964. Fishes of some South Carolina coastal plain streams.
Quarterly Journal of the Florida Academy of Science, 27:31-54.
Loyacano, H.A. 1975. A list of freshwater fishes of South Carolina.
Bulletin No. 580. South Carolina Agricultural Experiment Station.
SOUTH DAKOTA
Bailey, R.M. and M.O. Allum. 1962. Fishes of South Dakota. Miscellaneous
Publications of the Museum of Zoology, University of Michigan. No. 119.
131 pp.
TENNESSEE
Etnier, D.A. and W.C. Starnes. 1993. The fishes of Tennessee. Univ.
Tennessee Press, Knoxville, TN. In Press.
TEXAS
Hubbs, C. 1972. A checklist of Texas freshwater fishes. Texas Parks and
Wildlife Department Technical Service 11:1-11.
Knapp, F.T. 1953. Fishes found in the fresh waters of Texas. Ragland Studio
and Lithograph Printing Company, Brunswick, Georgia, TX. 166 pp.
UTAH
Sigler, W.F. and R.R. Miller. 1963. Fishes of Utah. Utah Game and Fish
Department. Salt Lake City, UT. 203 pp.
VERMONT
MacMartin, J.M. 1962. Vermont stream survey 1952-1960. Vermont Fish and
Game Department, Montpelier, VT. 107 pp.
VIRGINIA
Jenkins, R.E. and N.M. Burkhead In Press. The freshwater fishes of
Virginia. American fisheries Society,Bethesda, MD.
188
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WASHINGTON
Wydoski, R.S. and R.R. Whitney. 1979. Inland fishes of Washington.
University of Washington Press. 220 pp.
WEST VIRGINIA
Denoncourt, R.R., E.G. Raney, C.H. Hocutt, and J.R. Stauffer, Jr. 1975. A
checklist of the fishes of West Virginia. Virginia Journal Science
26(3):117-120.
Hocutt, C.H., R.F. Denoncourt, and J.R. Stauffer, Jr. 1979. Fishes of the
Gauley River, West Virginia. Brimleyana 1:47-80.
WISCONSIN
Becker, G.C. 1983. Fishes of Wisconsin. University of Wisconsin Press,
Madison, WI. 1052 pp.
WYOMING
Baxter, G.T. and J.R. Simon. 1970. Wyoming fishes. Wyoming Game and Fish
Department. Bulletin No. 4, Cheyene, WY. 168 pp.
CANADA
McPhail, J.D. and C.C. Lindsey. 1970. Freshwater fishes of northwestern
Canada and Alaska. Bulletin No. 173. Fisheries Research Board of
Canada. 381 pp.
Scott, W.B. and E.J. Crossman. 1973. Bulletin No. 1984. Freshwater fishes
of Canada. Fisheries Res. Board Canada. 866 pp.
Walters, V. 1955. Fishes of western Arctic America and Alaska. Bulletin
of the American Museum of Natural History 106:259-368.
EASTERN CANADA
Hubbs, C.L. and K,F. Lagler. 1964. Fishes of the Great Lakes Region.
University of Michigan Press, Ann Arbor, Michigan. 213 pp.
McAllister, D.E. and B.W. Coad. 1974. Fishes of Canada's National Capital
Region. Special Publication 24. Fisheries and Marine Service. 200 pp.
ALBERTA
Paetz, M.J. and J.S. Nelson. 1970. The fishes of Alberta. Queen's Printer,
Edmonton, Alberta. 282 pp.
BRITISH COLUMBIA
Carl. G.C., W.A. Clemens, and C.C. Lindsey. 1967. The freshwater fishes of
189
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British Columbia. Fourth edition. Handbook No. 5. British Columbia
Provincial Museum. 192 pp.
Hart. J.L. 1973. Pacific fishes. Second edition. Bulletin No. 180.
fisheries Research Board of Canada. 740 pp.
MANITOBA
Fedoruk, A.N. 1969. Checklist and key of the freshwater fishes of Manitoba.
Manitoba Department of Mines and Natural Resources, Canada Land
Inventory Project. 98 pp.
Hinks, D. 1943. The fishes of Manitoba. Manitoba Department of Mines and
Natural Resources. 101 pp.
NEW BRUNSWICK
Gorhatn, S.W. 1970. Distributinal checklist of the fishes of New
Brunswick. Saint John, New Brunswick. 32 pp.
Scott, W.B. and E.J. Grossman. 1959. The freshwater fishes of New Brunswick.
A checklist with distributional notes. Contribution No. 51. Royal
Ontario Museum, Division of Zoology and Palaeontology. 37 pp.
NORTHWEST TERRITORIES
Stein, J.N., C.S. Jessop, T.R. Porter, and K.T.J. Chang-Kue. 1973. An
evaluation of the fish resources of the Mckenzie River Valley as related
to pipeline development. Volume 1. Report 73-1. Information Canada
Catalogue Number FS37-1973/1-1, Environmental-Social Committee Northern
Pipelines, Task Force on Northern Development. 122 pp.
NOVA SCOTIA
Gil hen, J. 1974. The fishes of Nova Scotia's lakes and streams. Nova Scotia
Museum, Halifax. 49 pp.
Livingston, D.A. 1951. The freshwater fishes of Nova Scotia. Nova Scotian
Institute of Science Proceedings. 23:1-90.
ONTARIO
MacKay, H.H. 1963. Fishes of Ontario. Ontario Department of Lands and
Forest. 360 pp.
Ryder, R.A., W.B. Scott, and E.J. Crossman. 1964. Fishes of Northern
Ontario, North of the Albany River. Life Sciences Contribution, Royal
Ontario Museum. 30 pp.
QUEBEC
Legendre, V. 1954. Key to game and commercial fishes of the Province of
190
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Quebec. First English edition. Quebec Department of Game and
Fisheries. 189 pp.
Masse, G. et J. Mongeau. 1974. Repartition Geographique des Poissons, leur
abondance relative et bathymetric de la region du Lac Saint-Pierre.
Service de 1'Amenagement de la Faune, Ministere du Tourisme, de la
Chasse et de la Peche. Quebec. 59 pp.
Melancon, C. 1958. Les Poissons de nos Eaux. Third edition. La Societe
Zoologique de Quebec. Quebec. 254 pp.
Mongeau, J., A. Courtemanche, G. Masse, et Bernard Vincent. 1974. Cartes
de repartition geographique des especes de poissons au sud du Quebec,
d'apres les inventaries ichthyologiques effectues de 1963 a 1972.
rapport Special 4, Faune du Quebec. 92 pp.
Mongeau J., et G. Masse. 1976. Les poissons de la region de Montreal, la
peche sportive et commerciale, les ensemencements, les frayeres, la
contamination par le mercure et les PCB. Service de TAmegagement de la
Faune, Ministere du Tourisme, de la Chasse et de la Peche, Quebec. 286.
SASKATCHEWAN
Symington, D.F. 1959. The fish of Saskatchewan. Conservation Bulletin No.
7. Saskatchewan Department of Natural Resources. 25 pp.
YUKON TERRITORY
Bryan, J.E. 1973. The influence of pipeline development on freshwater
fishery resources of northern Yukon Territory, Aspects of research
conducted in 1971 and 1972. Report No. 73-6. Information Canada
Catalogue Number R72-9773. Environmental-Social Committee Northern
Pipelines, Task Force on Northern development. 63 pp.
GENERAL
Grossman, E.J. and H.D. VanMeter. 1979. Annotated list of the fishes of
the Lake Ontario watershed. Technical Report 36. Great Lakes Fishery
Commission, Ann Arbor, MI.
Eddy, S. and T. Surber. 1947. Northern fishes with special reference to
the Upper Mississippi Valley, 2nd edition. University of Minnesota
Press. Second edition. Minneapolis, MN. 267 pp.
Hocutt, C.H. and E.O. Wiley. 1986. The zoogeography of North American
freshwater fishes. John Wiley and Sons, NY.
Hubbs, C.L. and K.F. Lagler. 1947. Fishes of the Great Lakes Region. The
Cranbrook Press, Bloomfield Hills, MI. 186 pp.
Jenkins, R.E., E.A. Lachner, and F.J. Schwartz. 1972. Fishes of the
191
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central Appalachian drainages: Their distribution and dispersal. In:
The Distributional History of the Biota of the Southern Appalachians.
Part III: Vertebrates (P. C. Holt, ed.), Research Division Monograph 4.
Virginia Polytechnic Institute and State University, Blacksburg, VA.
Kuehne, R.A. and R.W. Barbour. 1983. The American darters. Univ. Kentucky
Press, Lexington, KY.
Lee, D.S., C.R. Gilbert, C.H. Hocutt, R.E. Jenkins, D.E McAllister, and J.R.
Stauffer, Jr. 1980. Atlas of North American freshwater fishes.
North Carolina Museum of Natural History, Raleigh, NC.
Metcalf, A.L. 1966. Fishes of the Kansas River system in relation to
zoogeogarphy of the Great Plains. Publication of the Museum of
Natural History, University of Kansas 17(3):23-189.
Miller, R.R. 1948. The cyprinodont fishes of the Death Valley system of
eastern California and southwestern Nevada. Miscellaneous Publication
of the Museum of Zoology. University of Michigan 68:1-55.
Miller, R.R. 1959. Origin and affinities of the freshwater fish fauna of
western North America. Zoogeography publication number 51. American
Association for the advancement of Science, Washington, DC.
Page, L.M. 1983. Handbook of darters. TFH Pub!., Neptune, NJ. 271 pp.
Page, L.M. and B.M. Burr. 1991. A field guide to freshwater fishes.
Houghton Miff!in Co., Boston, MA. 432 pp.
Rostlund, E. 1952. Freshwater fish and fishing in native North America.
University of California Geography Publications 9:1-313.
Seehorn, M.E. 1975. Fishes of southeastern national forests. Proceedings
29th Annual Conference Southeastern Association Game Fish Commission,
pp 10-27.
Sigler, W.F. and J.W. Sigler. 1987. Fishes of the Great Basin. Univ.
Nevada Press, Reno, NE. 425 pp.
Soltz, D.L. and R.J. Naiman. 1978, The natural history of native fishes in
the Death Valley system. Natural History Museum of Los Angeles County.
Science Series 30:1-76.
Tomelleri, J.R. and M.E. Eberle. 1990. Fishes of the central United States.
Univ. Press of Kansas, Lawerence, KS. 432 pp.
8.15 Agencies Currently Using or Evaluating Use of the IBI and Iwb for Water
Quality Investigations
1. Alabama Geological Survey
2. Illinois Environmental Protection Agency
192
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3. Iowa Conservation Commission
4. Kansas Department of Wildlife and Parks
5. Kansas Department of Health and Environment
6. Kentucky Cabinet for Natural Resources and Environmental Protection
7. Nebraska Department of Environmental Control
8. North Carolina Division of Environmental Management
9. Ohio Environmental Protection Agency
10. Oklahoma State Department of Health
11. Tennessee Valley Authority
12. U.S. EPA Region I
13. U.S. EPA Region II
14. U.S. EPA Region V
15. Vermont Department of Environmental Conservation
16. Wisconsin Department of Natural Resources
17. Indiana Department of Environmental Management
18. Arizona Department of Game and Fish
8.16 Ohio EPA Fish Index of Biotic Integrity (IBI), Modified Index of Well-
Being (Iwb), and Qualitative Habitat Evaluation Index (QHEI)
8.16.1 The principal methods for determining the overall fish community
health and well-being used by the Ohio EPA are the Index of Well-Being (Iwb)
developed by Gammon (1976), and modified by Ohio EPA (see Ohio EPA, 1987b,
1991), the Index of Biotic Integrity (IBI) developed by Karr (1981), and the
qualitative habitat evaluation index (QHEI) developed by Rankin (1989). The
Iwb is based on structural attributes of the fish community, and the IBI
incorporates functional characteristics. The fish technique used by Ohio EPA
to obtain fish relative abundance and distribution data is pulsed direct
current (D.C.) electrofishing. Depending on the type of habitat sampled, six
sampling methods currently being used are: (1) boat-mounted electrofishing -
straight electrode array (2) boat-mounted electrofishing - circular electrode
array, (3) boat longline - riffle method; (4) Sportyak generator unit (5)
longline generator unit, and (6) Backpack electrofishing - battery unit. Fish
data collected with these devices are used for the purpose of calculating the
Index of Biotic Integrity (IBI) and Modified Index of Well-Being (Iwb) scores
from which aquatic life use attainment and water quality are determined.
Figure 13 is a flowchart of the biosurvey approach for fish bioassessment used
193
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I. Reference Sites - Select & Sample
II. Calibrate multi-metric indices (IBI, ICI)
lf>
Ul
0
LJ
a
CO
i
O
10 100
DRAINAGE AREA (SQ Ml)
1000
III. Fully calibrated index - differentiate site
types for fish; statewide for invertebrates.
IBI - calibrated for use in Ohio for Wading Sites.
Category
Species
Comp-
osition
Trophic
Comp-
osition
Fish
Condition
IBI Metric
# of Species
# of Darters
i of Sunfish
# of Suckers
It of Intok-ranls
<100 Sq, Mi,
>100Sq. Mi.
% Tolerants -
% Omnivores
% Inseciivores
<30 Sq, Mi.
>30 Sq. Mi.
% Top Carnivores
# of Individuals
% Simple Litho.
% DELTs
Metric Score
ill
Varies with drainage area
Varies with drainage area
>3 2-3 <2
Varies with drainage area
>5 3-5 <3
Varies with drainage area
Varies with drainage area
>19 19-34 >34
Varies with drainage area
>55 26-55 <26
>5 1-5 <1
>750 200-750 <200
>36 18-36 <18
<0.1 0.1-1.3 <200
IV, Evaluate reference site score distribution-
examine for ecoregion differences.
£ 6 0
EC
'S SO
O 4 0
3 30
Q Z°
i
— i o
Rofarence Results - Wading Sites
(~ 1
T
_i (.
HELP EOLP IP ECBP WAP
V. Derive numerical biocriteria for each aquatic life
use designation as defined in the Ohio WQS.
VI. Use biocriteria in ambient assessments.
Middle Scloto R. 1979 vs 1988
140
13O 12O 11O 1OO
RIVER MILE
Figure 13. Flowchart of biosurvey approach for fish bioassessment used by Ohio EPA (1991).
-------
by Ohio EPA. Figure 14 is an example of a fish data sheet constructed for
immediate entry into a computer data base,
8.16.2 Ohio EPA (1989) also collects data for a general qualitative habitat
evaluation (Figure 12} for calculating the Qualitative Habitat Evaluation
Index (QHEI) developed by Rankin (1989). The QHEI is designed to provide an
empirical, quantified evaluation of the general lotic maerohabitat
characteristics that are important to fish communities. A detailed analysis
of the development and use of the QHEI is found in Rankin (1989).
8.16.3 For details of specific Ohio EPA field and laboratory methods for fish
bioassessment (e.g., sampling site selection, fish sampling procedures, field
counting and weighing procedures, handling preserved specimens, data handling
and analysis), one should consult Ohio EPA (1987a, 1987b, 1989, 1990b).
195
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Fish Data Sheet
_. . . — Collector/Recorder
Field Crew:
of
River/Stream:
Date:
River Code:
RM:
Distance:
~ Sampler Type: .„...
LJepin: „ __,|M .„,„„ _
_» —
™" Uata Source:
Location:
Time fished: '
" Tola! Seconds
Observed Flow! . ,„,. , ,
Number of Species: _____
AnomaKas. A-andtcr Mm, B-bladt spot; C-teecfes. Wetamdiss; E-mati Ira. Rwgus: L-tosions; U-muRipK CELT anomai*$,
P-parasitas; Y-popeys; S-emaoatsO; W-$wiriad scales: T-uncra, Z-effw/. (H-Haavy; L-OgW v» comdned wiffi anomalies A, 8. and Cj
SPECIES
f WEIGHEC
TOTAL
fQUIfTEO
i
WEIGHT (GRAMS)
nzz
«^IZJ
Mass Weighing Twu .
Convention: «,!«*« M
ANOUALIES
536 Ml)-
• "Mlllll^
MHHWBM HjRtjW
H«ofwd
Figure 14. Example of Ohio EPA (1991) field data sheet constructed for
immediate entry into a computer data base.
196
-------
Page
of
Anxnaliss; A-anehor worm. B-Uack spot: C-tMcta: Mffcnwtw: E-*rod«d fas, Mungus; L-tovora; M-mdtipte OELT anomalies, N-blno.
P-patasiias; Y-popeya; S*«macated; W-swirtrt icstos; T-iumon; Z-ofw/. (H-H«*y; l-Ught u« csmSnsd mm anomalies A. B. and C]
SPECIES
f WEIOKCC
TUfmL
CpUHTED
!
1
!
1
WEIGHT (GR£MS|
Mass Weighing fa|gj „.
Convention; «,.,..,, ...
• — 536 fll)-
ANOMALIES
— • i ,.._ Nuntm
Wasted
Figure 14. Example of Ohio EPA (1991) field data sheet constructed for
immediate entry into a computer data base (continued).
197
-------
8.17 Literature Cited
Angemeier, P.L. 1983. The importance of cover and other habitat features
to the distribution and abundance of Illinois stream fishes. Ph.D.
Dissertation, University of Illinois, Urbana, IL.
Angemeier, P.L, and J.R. Karr. 1986, Applying an index of biotic integrity
based on stream fish communities: Considerations in sampling and
interpretation. N. Am. J. Fish. Manag. 6:418-429.
Ball, J. 1982. Stream classification guidelines for Wisconsin. Wisconsin
Department of Natual Resources Technical Bulletin. Wisconsin Department
of Natural Resources, Madison, HI.
Barbour, M.T. and J.B. Stribling. 1991. Use of habitat assessment in
evaluating the biological integrity of stream communities. EPA-440/5-
91-005. In: Biolical criteria: Research and Regulation, 1991.
Proceedings of a Symposium, U.S. Environmental Protection Agency, Office
of Water, Washington, DC. pp. 25-38.
Bickers, C.A., M.H. Kelly, J,M. Levesque, and R.L. Hite. 1988. User's guide
to IBI-AIBI-Version 2.01 (A basic program for computing the index of
biotic integrity with the IBM-PC). State of Illinois, Environmental
Protection Agency, Marion, IL.
Bond, C.E. 1988. Department of Fisheries and Wildlife, Oregon State
University, Corvallis. Personal Communication.
Bramblett, R.G. and K.D. Fausch. 1991. Variable fish communities and the
Index of Biotic Integrity in a western great plains river. Trans. Amer.
Fish, Soc. 120:752-769.
Cairns, J., Jr. and R.L. Kaesler. 1971. Cluster analysis of fish in a
portion of the Upper Potomac River. Trans. Am, Fish. Soc. 100:750-756.
Dimick, R.E. and F. Merryfield. 1945. The fishes of the Willamette River
system in relation to pollution. Engineering Experiment Station
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index of biotic integrity based on stream fish communities. Trans. Am.
Fish. Soc. 113:39-55
Funk, J.L. 1957. Movement of stream fishes in missouri. Trans. Am. Fish.
Soc. 85:39-57.
Gammon, J.R. 1976. The fish populations of the middle 340 km of the Wabash
River. Purdue Univ. Water Resources Res. Cen. Tech. Rep 86. 73 pp.
Gammon, J.R. 1980. The use of community parameters drived from
electrofishing catches of river fish as indicators of environmental
198
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quality, in seminar on water quality management tradeoffs. Report No
EPA-905/9-8Q-009. U.S. EPA, Washington, DC.
Gammon, J.R. 1989. Personal communication, Department of Biological
Sciences. DePauw University, Greencastle, IN.
Gammon, J.R., A, Spacie, J.L. Hamelink, and R.L. Kaesler. 1981. Role of
electrofishing in assessing environmental quality of the Wabash
River. In: Ecological assessments of effluent impacts on communities
of indigenous aquatic organisms. J. M. Bates and C. I. Weber, eds, STP
730, pp. 307-324. American Society of Testing and Materials,
Philadelphia, PA.
Gammon, D.B. Halliwell, P.L. Angemeier, D.J, Orth. 1988. Regional
applications of index of biotic integrity for use in water resource
management. Fisheries 5:12-20.
Gauch, H., Jr. 1982. Multivariate analysis in community ecology. Cambridge
Univ. Press, NY.
Gerking, S.D. 1959. The restricted movement of fish populations. Biol.
Review 34:221-242.
Hendricks, M.L., C.H. Hocutt, and J.R. Stauffer, Jr. 1980. Monitoring of
fish in lotic habitats. In: Biological Monitor of Fish, C. H. Hocutt
and J.R. Stauffer, Jr., eds. D. C. Heath Co., Lexington, MA.
Hill, M.O. 1979. DECORANA: a fortran program for detrended correspondence
analysis and reciprocal averaging. Cornell University, Ithaca, NY.
Hill, J. and G.D. Grossman. 1987. Home range estimates for three North
American stream fishes. Copeia 1987:376-380.
Hocutt, C.H. 1981. Fish as indicators of biologic integrity. Fisheries
6(6):28-31.
Hughes, R.M. 1985. Use of watershed characteristics to select control streams
for estimating effects of metal mining wastes on extensively distrurbed
streams. Environ. Manage. 9:253-262,
Hughes, R.M., J.H. Gakstater, M.A. Shirazi, and J.M. Omernik. 1982. An
approach for determining biological integrity in flowing waters. In:
In place resource inventories: Principles and practices. Proceedings
of a National Workshop, T. B. Brann, ed. Society of American Foresters,
Bethesda, MD.
Hughes, R.M. and J.R, Gammon. 1987. Longitudinal changes in fish assemblages
and water quality in the Willamette River, Oregon. Trans. Am. Fish.
Soc. 116(2):196-209.
Hughes, R.M., D.P. Larsen, and J.M. Omernik. 1986. Regional reference sites:
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A method for assessing stream potentials. Environ. Manage. 10:629-635.
Hughes, R.M., E. Rexstad, and C.E. Bond. 1987. The relationship of
aquatic ecoregions, river basins, and physiographic provinces to the
ichthyogeographic regions of Oregon. Copeia 1987:423-432.
Hughes, R.M. and D.P. Larsen. 1988. Ecoregions: an approach to surface
water protection. J. Water Pollut. Control Fed. 60:486-493.
Judy, R.D., Jr., P.N. Seeley, T.M. Murray, S.C. Svirsky, M.R. Whitworth, and
L.S. Ischinger. 1984. Technical Report, Initial Findings: Vol. 1 of
1982 National Fisheries Survey. Report No. FWS/OBS-84/06. U. S. Fish
and Wildlife Service, Fort Collins, CO.
Karr, J.R. 1981. Assessment of biotic integrity using fish communities.
Fisheries 6:21-27.
Karr, J.R., K.D. Fausch, P.L. Angermeier, P.R. Yant, and I.J. Schlosser.
1986. Assessing biological integrity in running waters: A method and
its rationale. Special Publication 5. Illinois Natural History Survey.
Kuehne, R.A. and R.W. Barbour. 1983. The American darters. University
Kentucky Press, Lexington, KY.
Larsen, D.P., J.M. Omernik, R.M. Hughes, C.M. Rohmm, T.R. Whittier, A.J.
Kinney, A.L. Gallant, and D.R. Dudley. 1986. The correspondence
between spatial patterns in fish assemblages in Ohio streams and aquatic
ecoregions. Environm. Manage. 10:815-828.
Larsen, D.P., D.R. Dudley, and R.M. Hughes. 1988. A regional approach for
assessing attainable water quality: An Ohio case study. J, Soil
Water Conserv. 43:171-176.
Leonard, P.M. and D.J. Orth. 1986, Application and testing of an index of
biotic integrity in small, cool-water streams. Trans. Amer. Fish. Soc.
115:404-414.
Lyons, J. 1992. Using the Index of Biotic Integrity (IBI) to measure
environmental quality in warmwater streams of Wisconsin. U.S.
Department of Agriculture, Forest Service, General Technical Report NC
149.
Matthews, W.J. 1986. Fish fauna! structure in an Ozark stream: Stability,
persistence, and a catastrophic flood. Copeia. 1986:388-397.
Matthews, W.J., D.J. Hough, and H.W. Robison. 1992. Similarities in fish
distribution and water quality patterns in streams of Arkansas:
Congruence of multivariate analyses. Copeia 2:296-305.
Miller, D.L., P.M. Leonard, R.M. Hughes, J.R. Karr, P.B. Moyle, L.H. Schrader,
B.A. Thompson, R.A. Daniels, K.D. Fausch, G.A. Fitzhugh, J.R. Gammon,
D.B, Halliwell, P.L. Angermeier, and D.J. Orth. 1988a. Regional
200
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applications of an Index of Biotic Integrity for use in water resource
management. Fisheries 5:12-20,
Miller, D.L., R.A. Daniels, and D.B. Halliwell. 1988b. Modification of an
Index of Biotic Integrity based on fish communities for streams of the
northeastern United States. Unpublished Manuscript.
Moyle, P.B. 1976. Inland fishes of California. University of California
Press, Berkeley, CA.
Nielsen, L.A. and D.L. Johnson, eds. 1983. Fisheries techniques. American
Fisheries Society, Bethesda, MD.
Ohio EPA. 1987a. Biological criteria for the protection of aquatic life:
Volume I. The role of biological data in water quality assessment.
Ohio Environmental Protection Agency, Ecological Assessment Section,
Division of Water Quality & Assessment, Ohio Environmental Protection
Agency, Columbus, OH.
Ohio EPA. 1987b. Biological criteria for the protection of aquatic life:
Volume II. User's manual for biological assessment of Ohio surface
waters. Ohio Environmental Protection Agency, Ecological Assessment
Section, Division Water Quality & Assessment, Columbus, OH.
Ohio EPA. 1987c. Appendix B: Development of fish community IBI metrics.
Appendix C: Modified Index of Well-Being (Iwb). In: Biological
criteria for the protection of aquatic life: Volume II: Users manual
for biological field assessment of Ohio surface waters. Ohio
Environmental Protection Agency, Ecological Assessment Section, Division
Water Quality Monitoring & Assessment, Columbus, OH.
Ohio EPA. 1989, Biological criteria for the protection of aquatic life:
Volume III. Standardized biological field sampling and laboratory
methods for assessing fish and macroinvertebrate communities. Ohio
Environmental Protection Agency, Ecological Assessment Section, Division
Water Quality Monitoring & Assessment, Ohio Environmental Protection
Agency, Columbus, OH.
Ohio EPA. 1990a. Compendium of biological results from Ohio rivers, streams,
and lakes: 1989 edition. Ecological Assessment Section, Division
Water Quality Planning and Assessment, Ecological Assessment Section,
Columbus, OH.
Ohio EPA. 199Qb. The use of biocriteria in the Ohio EPA surface water
monitoring and assessment program. Ohio Environmental Protection
Agency, Ecological Assessment Section, Division Water Quality Planning
and Assessment, Ecological Assessment Section, Columbus, OH.
Ohio EPA. 1990c. Fish evaluation group safety manual. Ohio Environmental
Protection Agency, Ecological Assessment Section, Division Water Quality
Planning and Assessment, Ecological Assessment Section, Columbus, OH.
201
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Ohio EPA. 1991. Ohio EPA outline of regional reference site approach to
deriving numerical biological criteria. 1991 MPCB Meeting: Region V.
Biocriteria Work Group. Division Water Quality Planning and
Assessment, Ecological Assessment Section, Columbus, OH.
Omernik, J.M. 1987. Ecoregions of the conterminous United States. Ann.
assoc. Am. Geograph. 77:118-125.
Omernik, J.M. and A.L. Gallant. 1988. Ecoregions of the upper midwest
states. EPA/600/3-88/037. U.S. Environmental Protection Agency,
Environmental research Laboratory, Corvallis, OR.
Osborne, L.L. and E,E. Hendricks. 1983. Streamflow and velocity as
determinants of aquatic insect distribution and benthic community
structure in Illinois. Water Resources Center, University of
Illinois, Report No!. UILU-WRC-83-183. U.S. Department of the
Interior, Bureau of Reclamation.
Oswood, M.E. and W.E. Barber. 1982. Assessment of fish habitat in
streams: Goals, constraints, and a new technique. Fisheries 7(3):8-ll.
Page, L.M. 1983. Handbook of darters. TFH Publication, Inc., Ltd.,
Neptune City, NJ.
Plafkin, J.L., M.T. Barbour, K.D. Porter, S.K. Gross, and R.M. Hughes.
1989. Rapjd bioassessment protocols for use in streams and rivers:
benthic macroinvertebrates and fish. EPA/440/4-89/001. Office of Water,
Assessment and Watershed Protection Division, U. S. Environmental
Protection Agency, Washington, DC.
Platts, W.S., W.F. Megahan, G.W. Minshall. 1983. Methods for evaluating
stream, riparian, and biotic conditions. General Technical Report INT-
138. U. S. Department of Agriculture, U. S. Forest Service, Ogden, UT.
Rankin, E.T. 1987. Ohio Environmental Protection Agency, Columbus, OH.
Personal communication.
Rankin, E.T. 1989. The qualitative habitat evaluation index (QHEI):
rationale, methods, and application. Ohio EPA, Ecological Assessment
Section, Division of Water Quality Planning & Assessment, P.O. Box 1049,
1800 WaterMark Drive, Columbus, OH.
Reynolds, J.B. 1983. Electrofishing. In: Fisheries Techniques. L. A.
Nielsen and D L. Johnson, eds. American Fisheries Society, Bethesda,
MD.
Rohm, C.M., J.W. Giese, and C.C. Bennett. 1987. Evaluation of an aquatic
ecoregion classification of streams in Arkansas. Freshwater Ecol.
4:127-140.
Ross, S.T., W.J. Matthews, and A.E. Echelle. 1985. Persistence of stream
fish assemblages: Effects of environmental change. Am. Nat. 126:24-40.
202
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Sanders, R.E. 1991. A 1990 night electrofishing survey of the upper
Ohio River Mainstem (RM 40,5 to 270.8} and recommendations for a long-
term monitoring program. Ohio Dept. Nat. Res. (ODNR), Division of
Wildlife, 1840 Belcher Dr., Columbus, OH.
Sanders, R.E. 1992. Day versus night electrofishing catches from near
shore waters of the Ohio and Muskingum Rivers. Ohio J. Sci. 93(3):In
Press.
Schrader, L.H. 1989. Use of the index of biotic integrity to evaluate the
effects of habitat, flow, and water quality on fish communities in three
Colorado front range rivers. Master's Thesis. Colorado State
University, Fort Collins, CO.
Scott, W.B. and E.J. Grossman. 1973. Freshwater Fishes of Canada.
Fisheries Resources Board of Canada, Bulletin 184.
Seber, G.A. 1982. The estimation of animal abundance. McMillan
Publishing, New York, NY.
Seber, G.A.F. and E.D. LeCren. 1967. Estimating population parameters
from catches large relative to the population. J. Anim. Ecol. 36:631
-643.
Seber, G.A.F. and J.F. Whale. 1970. The removal method for two and three
samples. Biometrics. 26:393-400.
Simon, T. 1990. Instream water quality evaluation of the upper Illinois River
basin using the Index of Biotic Integrity. EPA-905/9-90-005. In: W.S.
Davis (ed.). Proceedings of the 1990 midwest pollution control
biologists meeting. U.S. Environmenta Protection Agency, Environmental
Division, Chicago, IL. pp. 124-142.
Simon, T. 1991. Development of index of biotic integrity expectations for
the ecoregions of Indiana. I. central corn belt plain. EPA-905/9-
91/025. U.S. Environmental Protection Agency, Environmental Services
Division, Monitoring and Quality Assurance Branch, Ambient Monitoring
Section, Chicago, IL.
Simpson, J.C. and R.L. Wallace. 198E. Fishes of Idaho. University Press
of Idaho, Moscow, ID.
Steedman, R.J. 1988. Modification and assessment of an index of biotic
Integrity to quantify stream quality in southern Ontario. Can J.
Fish. Aquat. Sci. 45:492-501.
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for generating population statistics from electrofishing data-user's
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Agriculture, U.S. Forest Service, Ogden, UT.
Wade, D.C. and S.B. Stalcup. 1987. Assessment of the sport fishery potential
203
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for the Bear Creek floatway: Bloloagical integrity of representative
sites, 1986. Report No. TVA/ONARED/AWR-87/30. Tennessee Valley
Authority, Muscle Shoals, AL.
Whittier, T.R., R.M. Hughes, and D.P. Larson. 1988. Correspondence between
ecoregions and spatial patterns in stream ecosystems in Oregon. Can. J.
Fish. Aquati. Sci. 45:1264-1278.
Wydoski, R.S. and R.R. Whitney. 1979. Inland fishes of Washington.
University of Washington Press, Seattle, WA.
Yoder, C.O., P.A. Albeit, and M.A. Smith. 1981. The distribution and
abundance of fishers in the mainstem Scioto River as affected by
pollutant loadings. Ohio EPA Tech. Rept. 81/3. Columbus. 118 pp.
204
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SECTION 9
FAMILY-LEVEL ICHTHYOPLANKTON INDEX METHODS1
9.1 Introduction
9.1.1 The early life history stages of fishes are recognized as the most
sensitive and vulnerable life stage (Blaxter, 1974; Moser et al., 1984; Weis
and Weis, 1989). The ability to document status and trends without
identifying most taxa to species has caused some doubt as to the relevance of
resolution abilities of using ichthyoplankton in bioassessment studies.
9.1.2 Although there are some reluctance to conduct further ichthyoplankton
studies detailed enough to answer water quality questions, investigators have
continued to gather important and useful knowledge on the early life stages of
fishes. A recent explosion in the amount and types of literature includes
documentation of nursery habitats (Goodyear et al., 1982), ecological early
life history notes (Simon and Wallus, 1989; Wallus, 1986; Wallus and Buchanan,
1989), taxonomic studies of regionally important systems (Auer, 1982; Holland
and Huston, 1983; Simon, 1990; Wallus et al., 1989), toxicological studies
using early life history stages (Norberg and Mount, 1983; Birge et al., 1985;
Simon, 1988), and effects of environmental pollution (Weis and Weis, 1989).
9.1.3 The purpose of the family-level ichthyoplankton index methods is to
present guidelines and an index for the use of ichthyoplankton in
bioassessment studies and for determining water quality. The use of a
qualitative collection method with a family-level taxonomic approach will
facilitate use without complicating logistics and level of effort. The
family-level index is based on three components: taxonomy, reproductive
guild, and abundance and deformity. Water quality managers, in addition,
could use this information to document reproduction, nursery habitats, and
backwater habitats not conventionally surveyed during routine adult fish or
marcoinvertebrate collection. The format and structure of the ichthyoplankton
index (I2) is modeled after the index of biotic integrity (IBI) using a
family-level approach. Since the proponents of the IBI recommend against use
of larval and juvenile stages in they analyses (Angermeier and Karr 1986; Karr
et al., 1986), the I2 can be an additional use of data collected during a
routine adult sampling event. Current knowledge on the identification of most
freshwater faunas are limited, however, a listing of appropriate references is
included in Table 1.
9.1.4 The loss of habitat through the accumulation of toxic chemicals in the
sediment, reduction of dissolved oxygen, and increase in siltation, is perhaps
the greatest obstacle to the protection of environmental quality the
environmentalist must face. Degradation by conventional nonpoint sources of
pollution have yet to be addressed, rather efforts have concentrated on point
sources. USEPA has spent two decades quantifying the effluent quality of
point source dischargers. With toxicity endpoints established in industrial
1Adapted from Simon (1989).
205
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TABLE 1. TAXONOMIC LITERATURE USEFUL FOR IDENTIFICATION OF LARVAL AND EARLY
JUVENILE NORTH AMERICAN FRESHWATER FISH (ALSO SEE SECTION 12,
BIBLIOGRAPHY, SUBSECTION 12.4.2 LARVAL AND IMMATURE FISHES)
Author(s) and Publication Date
Region
Auer, 1982
Colton and Marak, 1969
Drewry, 1979
Elliott and Jimanez, 1981
Fish, 1932
Fritzsche, 1978
Hardy, 1978a
Hardy, 1978b
Holland and Huston, 1983
Hogue et al., 1976
Johnson, 1978
Jones et al., 1987
Lippson and Moran, 1974
Mansueti and Hardy, 1967
Martin and Drewry, 1978
May and Gasaway, 1967
McGowen, 1984
McGowen, 1989
Great Lakes Basin, emphasis Lake
Michigan
Northeast Coast, Black Island to Cape
Sable
Great Lakes Region
Beverly Salem Harbor Area, Massachusetts
Lake Erie
Mid-Atlantic Bight (Chaetodontidae
through Ophidiidae)
Mid-Atlantic Bight (Aphredoderidae
through Rachycentridae)
Mid-Atlantic Bight (Anguillidae throuygh
Syngnathidae)
Upper Mississippi River
Tennessee River
Mid-Atlantic Bight (Carangidae through
Ephippidae)
Mid-Atlantic Bight (Acipenseridae
through Ictaluridae)
Potomac River Estuary
Chesapeake Bay Region
Mid-Atlantic Bight (Stromateidae
through Ogcocephalidae)
Oklahoma, Canton Reservoir
South Carolina, Robinson Impoundment
North Carolina Piedmont Impoundment
206
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TABLE 1. TAXONOMIC LITERATURE USEFUL FOR IDENTIFICATION OF LARVAL AND EARLY
JUVENILE NORTH AMERICAN FRESHWATER FISH (CONTINUED) (ALSO SEE
SECTION 12, BIBLIOGRAPHY, SUBSECTION 12.4.2 LARVAL AND IMMATURE
FISHES)
Author(s) and Publication Date
Region
Scotton et al., 1973
Snyder, 1981
Sturm, 1988
Taber, 1969
Wall us et al., 1989
Wang, 1981
Wang and Kernehan, 1979
Delaware Bay Region
Upper Colorado River System, Colorado
Alaska
Oklahoma and Texas, Lake Texoma
Ohio River basin, emphasis on
Tennessee and Cumberland drainages
Sacramento-San Joaquin Estuary and
Moss Landing Harbor Elkhorn Slough, CA
Delaware Estuary
207
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and municipal permits, attention must be focused on instream degradation
through chronic exposure to ambient residents.
9.1.5 The effort to combine a community approach for addressing these issues
has been accomplished in adult fish (Karr, 1981; Karr et a!., 1986),
macroinvertebrates (Plafkin et al., 1989), and now with ichthyoplankton here
in this section. Karr and colleagues have described in detail the rationale
for this overall approach. The reader is referred to their documentation for
further reading rather than repeating their rationale (Karr et al. 1986). In
this Section details are provided for the scoring and information of an
ichthyoplankton index using a community based approach.
9.1.6 The need to look at various trophic levels in the analysis of
environmental degradation, through biological integrity, is difficult to
explore in insects due to taxonomic and limited ecological information. In
fishes, ontogenetic shifts during development not only is apparent in
morphological changes (Fuiman and Corazza, 1979), but also niche shifts
(George and Hadley, 1979; Brandt, 1986). The early life stages of fishes
often documents the use of habitats by endangered or rare species when the
adults can frequently not be found. The protection of these important
habitats require further consideration in protection of species diversity.
9.1.7 The I2 is an additional tool which can be concurrently conducted using
IBI type techniques, and the method may prove useful in both lotic and lentic
habitats. The difficulty in assessing lentic habitats is the inability of
species to recolonize closed systems. Field evaluations of both habitat types
are necessary prior to further evaluation of the method.
9.1.8 The implications of data quality depends on the calibration of the
metrics and collection of a representative sample (Davis and Simon, 1988).
Every effort should be made to incorporate quality assurance checks into
standard operating procedures and data analysis. Further refinement of
techniques and interpretation will become apparent with increases in knowledge
of a balance aquatic environment especially as recruitment success and early
life history states of fishes are influenced.
9.1.9 Interpretation of the I2 follows that previously established by the
IBI. The use of a three tiered scoring criteria, 5, 3, and 1, are assigned to
each metric depending on whether it approximates, deviates somewhat from, or
deviates strongly from the value expected at the least impacted ecoregion
reference site. The sampling site is then assigned to one of six quality
classes based on the sum total of the eleven metric ratings. The highest
score, 55, indicates a site without perturbation and deviations decline
proportionally. The qualitative ratings and descriptions of Karr (1981) range
from excellent to very poor (Table 2). These similar integrity classes and
attributes have been appropriately scaled for the I2 bases on those of Karr et
al. (1986).
9.1.10 Finally, although the level of discernment of taxa to a species level
would be highly desired, the taxonomic literature is unable to support this
level currently. The family level of discernment will reduce confusion among
novices using the techniques, provided a high level of reproducability, and
208
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TABLE 2. TOTAL ICHTHYOPLANKTON INDEX (I2) SCORES, INTEGRITY CLASSES, AND
ATTRIBUTES (MODIFIED FROM KARR, 1981)
Total I2 Score Integrity
(Sum Of 11 Metrics) Class Attributes
53-55 Excellent Comparable to the best situations
without human disturbance; all
regionally expected taxa for habitat,
stream size, and ecoregion, including
the most intolerant forms; balanced
guild structure and reproduction.
44-48 Good Species richness somewhat below
expectations, especially due to loss
of the most intolerant forms; some
taxa are present with less than
optimal abundances; guild structure
indicates signs of some stress.
37-40 Fair Signs of additional deterioration
include loss of intolerant forms,
skewed dominance, and guild
structure. Reduction in simple
lithophils and in mean generation
time.
26-31 Poor Dominated by r-strategists, tolerant
forms and pioneer species. Increase
in guild A.I, and in deformities or
teratogenic fish.
11-20 Very Poor Few fish present, lack of successful
reproduction in any guild, deformed
or teratogenicity frequently
observed.
No Fish Repeated sampling finds no fish.
209
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subsequently data quality assurance through accuracy. As an increase in the
ecological requirements and taxonomic literature become available, a more
sensitive analyses will be possible. Stimulation of single species and
comparative larval descriptions and species reproductive characterization
should receive higher priority among researchers in the field.
9.2 Hethods and Materials
9.2.1 Sampling and Requirements
9.2.1.1 The objectives of the I2 are to provide a rapid screening method
using a single collection event to determine effects of water quality on
reproduction and the early life stages of fishes. Collection of a
representative sample of ichthyoplankton requires a variety of gear types, and
geographical, spatial and temporal considerations. The greater the stream
complexity, the greater the distance needed to be sampled; e.g., a second
order stream should be surveyed approximately 100 m, while a good rule of
thumb is fifteen times the river width or two habitat cycles (Gammon et al.,
1981; Karr et al., 1986). Reproduction by fishes occurs within a smaller
habitat scale than adult species occurrence. Fishes may rely on a broader
area for foraging and etching out an existence, however, only specialized
"selecf'habitats are utilized for reproduction and serve as a nursery habitat.
Because of patchy distribution of eggs and larvae a large enough area needs to
be investigated to determine local use of a particular stream reach.
9.2.2 Gear Types
9.2.2.1 The more complex the environment the more numerous and sophisticated
are equipment needs. The most typical equipment used for collection of larval
fishes include, plankton nets; seines, dip nets, and sweep nets; light traps;
and push nets and benthic sleds. Snyder (1983) provides documentation on
rationale and use of most of the above equipment. Light traps can be
constructed for lentic (Faber, 1981; 1982), and lotic waters (Muth and Haynes,
1984), and information on the use of the equipment can be determined from
references contained therein. Push nets and benthic sleds are described by
Tuberville (1979) and Burch (1983). Also, see Section 4, Sample Collection
for Analysis of the Structure and Function of Fish Communities.
9.2.3 Geographical Considerations
9.2.3.1 Landscape differences have long been recognized, and methods to
differentiate between various scales have been attempted using zoogeographical
realms, biomes, and most recently ecoregions. The ecoregions concept is the
most consistent means if evaluating community composition for a water quality
based approach. Omernik (1987) defined the conterminous United States into a
series of smaller discrete units. Aquatic biological characterization using
this approach has been completed for adult fish and macroinvertebrates in
several States including Ohio (Larsen et al., 1986; Ohio EPA, 1987), Arkansas
(Bennett et al., 1987; Geise and Keith, 1988), North Carolina (Penrose and
Overton, 1988), and Vermont (Langdon, 1988),
9.2.4 Spatial Considerations
210
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9.2.4.1 Riffles of rapid flow areas are not the most likely places to
encounter larval or juvenile fishes, rather the head of a pool, side margin of
a channel and backwater areas are preferred. A representative larval sample
should be collected from all available habitats within a stream reach. For
example, a large river sample should consist of various depth fractions from
the main channel, main channel border, side border and backwaters. Low flow
areas will reveal higher diversity of taxa while the remaining large river
species will be collected while drifting in the main channel (Simon, 1986a).
These diverse areas should be pooled for an overall evaluation of the site
while each component habitats, "relative value", can be quantitatively
assessed for its contribution to the whole. Creeks, stream, and small rivers
will require fewer areas to comprise a representative sample, however, any
reduced flow or eddy area will be in need of sampling within a given location.
Ideal habitats include those with submerged and emergent aquatic macrophytes,
overhanging bank vegetation and roots.
9.2.5 Temporal Considerations
9.2.5.1 Numerous reports and journal articles have documented spawning
temperature requirements of various faunas. In order to collect a
representative sample from a particular location, familiarity with the
reproductive literature and selection of appropriate sample times are
necessary. For example, in the midwest the earliest spawning fishes initiate
spawning under the ice, with larval emergence and hatching immediately after
ice-out during late March and early April. The last species to initiate
spawning are usually finished by mid-July with a majority of species spawning
during June (Simon, 1986a). Ichthyoplankton and early juvenile sampling
should be initiated in the midwest, no sooner than mid-June and no later than
the end of September to ensure collection of a representative sample.
9.2.5.2 The use of different gear types will facilitate collection of
families which are earlier spawning, e.g. percids, cottids, salmonids, and
catostomids. Due to north to south temperature clines, and east to west
rainfall differences, species will cue on spawning earlier in the south and
west and later in the north and east for the same species. Sampling needs to
be adjusted accordingly.
9.2.5.3 Equally important is die! differences in specimen collection.
Numerous studies have documented significant differences between dusk and
sunset, daylight, and night sampling. The general pattern is the more turbid
the water body the less likely diel affects will be a problem. When one
decides to sample, is not as important as it is for them to be consistent.
Safety considerations and study objectives may not deem night sampling
necessary. However, light trap use, set up using an automatic timing device
may enable night time sampling without the inconvenience and danger. This
method has successfully been used by Alabama Power on the Tallapoosa River.
9.2.5.4 Since much of the North American fauna is incompletely described
(Simon, 1986b), use of the index is limited to a family approach until the
taxonomic literature facilitates species specific recognition. The eleven I2
metrics are based on three broad categories. Metrics are organized into
taxonomic composition, reproductive guild, and abundance, generation time and
211
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deformity categories. No single metric is always a reliable indicator of
degradation, however, relative sensitivity is determined by region, scale, and
application.
9.2.5.5 The metrics will react differentially based on the type of
perturbation. For example, if contaminated sediments are suspected, the
proportion of lithophils and number of sensitive families should decline
depending on the magnitude of the impact, while equitability and perhaps
deformity should increase.
9.2.5.6 The remainder of this section provides information, justification and
rationale behind each of the I2 metrics (Table 3). Additional refinement may
be necessary to meet the objectives of the investigators study.
9.2.5.7 Taxonomic Composition. This category is useful for assessing family
diversity and community richness. The current level of taxonomy requires that
discussion be limited to a family level but future use of the index may make
this a species specific approach. Expectations should be determined for
various stream size and calibrated by equipment based on information presented
in Fausch et al. (1984). Taxa diversity has been determined to be the best
sole indicator of "good" water quality. Sensitive families such as percids,
cottids. ictalurids, and others listed in Table 4, are useful for determining
the extent of impact to sediments and nursery habitats. Finally, dominance of
tolerant species increase proportionally to environmental degradation.
9.2.6 Metric 1. Total Number of Families. The fluctuation in number of
families of an ecoregion increased with stream order. If the same order
stream, in the same ecoregion, with similar habitat cycles were sampled, then
reduction in numbers of families would correspond to environmental
degradation. A number of investigators have determined number of taxa is the
single most important metric which highly correlates with more pristine water
quality (Ohio EPA, 1987; Davis and Lubin, 1989; Plafkin et al., 1989).
9.2.7 Metric 2, Number of Sensitive Families. Certain families of
freshwater fish are sensitive to degradation, particularly as a result of
reproduction requirements and early life ecology (Table 4). Families such as
Percidae, Cottidae, and Salmonidae are intolerant to siltation and low
dissolved oxygen. Sediment contamination due to toxins and low dissolved
oxygen inhibits most benthic families (e.g., Ictaluridae). Reduction in
habitat quality (e.g., channelization, thermal inputs, reservoir flooding)
reduces Catostomidae, Centrarchidae, Cyprinidae, and Fundulidae. Sensitive
families should be restricted to those most sensitive to low dissolved oxygen,
toxic chemicals, siltation, and reduced flow. Karr et al. (1986) suggested
that species sensitive to habitat degradation, especially siltation, are most
likely to be identified as intolerant.
9.2.8 Metric 3. Equitability/Dominance. As water quality declines certain
taxa tend to become increasingly abundant (Karr et al., 1986). Also, species
defined as r-strategists tend to inundate the environment with early life
phases (MacArthur, 1957; MacArthur and Wilson, 1967). The strategy to produce
large numbers of young are indicative of "pioneer" species which are
attempting to colonize perturbed areas. In habitats with least impacted
212
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TABLE 3. METRICS USED TO ASSESS ICHTHYOPLANKTON COMMUNITIES FROM FRESHWATERS
OF NORTH AMERICA
ScoringCriteria
Category
Metric
Taxonomic Composition
1. Total Number of Families
2, Number of Sensitive Families
3. Equitability/Dominance
4. Family Biotic Index
Reproductive Guild
5. % Non-guarding Guild A.I and A.2
6. % Guarding Guild B.I and B.2
7. % Bearers Guild C.I and C.2
8. % Simple Lithophil Mode Reprod.
Abundance, Generation Time,
and Deformity
9. Catch per Unit Effort
10. Mean Generation Time
11. % Deformity or Teratogenicity
Drainage Size and Ecoregion Dependent
Drainage Size and Ecoregion Dependent
>0.8-1.0 >0.6-0.8 0-< 0.6
0-4.5 >4.5-7.5 >7.5-10
Drainage Size and Ecoregion Dependent
Drainage Size and Ecoregion Dependent
Drainage Size and Ecoregion Dependent
Drainage Size and Ecoregion Dependent
Drainage Size and Ecoregion Dependent
Drainage Size and Ecoregion Dependent
>2-5%
213
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TABLE 4. SENSITIVITIES, MEAN GENERATION TIME, AND REPRODUCTIVE GUILD
CHARACTERISTICS OF 34 NORTH AMERICAN FRESHWATER FISH FAMILIES
Family
Petromyzontidae
Acipensideridae
Polyodontidae
Lepisosteidae
Amiidae
Anguillidae
Clupeidae
Hiodontidae
Salmonidae
Osmeridae
Umbridae
Esocidae
Characidae
Cyprinidae
Catostomidae
Cobitidae
Ictaluridae
Claridae
Amblyopsidae
Aphredoderidae
Percopsidae
Gadidae
Oryzintidae
Cyprinodontidae
Fundulidae
Poeciliidae
Atherinidae
Gasterosteidae
Moron idae
Centrarchidae
Elassomatldae
Percidae
Sciaenldae
Cichlidae
Cottidae
Sensitivity
Moderate
Moderate
Intolerant
Tolerant
Tolerant
-
Moderate
Intolerant
Intolerant
Moderate
Tolerant
Moderate
Moderate
Moderate
Intolerant
Intolerant
Intolerant
Tolerant
Intolerant
Tolerant
Moderate
Moderately
Tolerant
Intolerant
Intolerant
Tolerant
Moderate
Tolerant
Intolerant
Intolerant
Intolerant
Intolerant
Moderate
Tolerant
Intolerant
Generation
Time1
Short/Moderate
Long
Long
Moderate
Moderate
Moderate
Short
Short/Moderate
Moderate/Long
Short
Short
Moderate
Short
Short
Moderate
Short
Moderate
Moderate
Short
Short
Short
Moderate/Long
Short
Short
Short
Short
Short
Short
Moderate
Moderate
Short
Short
Moderate
Moderate
Short
FBI2
3
I
2
4
8
3
6
4
1
5
9
6
5
6 A.I,
4
4
3
10
4
8
7
5
7
2
5
8
3
9
6
5
3
0 A.I,
4
7
0
Reproductive
Guild
A.I
A.I
A.I
A.I
B.2
A.I
A.I
A.I
A.I
A.I
A.I
A.I
A.I
A. 2, B.I, B.2
A.I, A. 2
A.I
B.2
A. 2
C.I
C.I
A.I
A.I
C.2
A.I, A. 2
A.I, A. 2
C.2
A.I
B.2
A.I
B.I
B.2
A. 2, B.I, B.2
A.I
B.2
B.2
Classified as short, moderate, and long appropriately scored 1, 3, 5,
respectively. A community mean is calculated by summing scores and dividing
by total number of families.
2Scored from 0 to 10.
organic enrichment.
The higher the score the greater the tolerance to
FBI = Family Biotic Index.
214
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environments, taxa tend to be equally distributed and more moderately
abundant. The Shannon diversity index and the measure of evenness are used to
determine quality environments which have balanced communities. These single
unit measures are not adequate in themselves to extrapolate excellent quality,
but they do determine increasing levels of disturbance, Equitability (Lloyd
and Ghelardi, 1964) is determined by comparing the number of families in the
sample with the expected number of families from a community which conforms to
the MacArthur broken stick model. MacArthurs' broken stick model is normally
higher than real diversity and is the ecologically maximum diversity
attainable (Washington, 1984). Equitability is measured by:
e = s'/s
where:
s = number of taxa in the sample,
s'= the tabulated value based on the Shannon diversity index
The diversity index is the d formulation of Lloyd, Zar, and Karr (1968), The
diversity index is:
d = C/N (N Iog10 N - E nf Iog10 nf)
where:
C = 3.321928,
N = total number of individuals in the ith taxa,
nf = total number of individuals in the ith taxa.
An example calculation and reproduction of Lloyd and Ghelardi's (1964)
table are included in Table 5 and are taken from USEPA (1973, 1990). As a
side note, if solely ichthyoplankton data sets are to be used excluding
juveniles, the following families need to be omitted: Clupeodae. Scianenidae,
and Osmeridae.
9.2.9 Metric 4. Family Biotic Index. Discussions with other
ichthyoplanktologists studying the ecological and taxonomic early life stages
of fishes suggest varying degrees of sensitivity exists between organic
pollution and perturbations such as sediment, degradation, siltation, low
dissolved oxygen, toxic chemicals, and flow reduction (Table 4). The
calculation of the Family Biotic Index (FBI) is modeled after Hilsenhoff's
(1988) modified biotic index which summarizes tolerances to organic pollution.
Tolerance values range between 0 to 10 for families and increase as water
quality decreases. The formula for calculating the Family Biotic Index is:
FBI = e x^./N
where:
Xj = total number of individuals within a taxon,
tj = tolerance value of a taxon,
N = total number of organisms in the sample.
215
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TABLE 5, THE DIVERSITY OF SPECIES, d, CHARACTERISTIC OF MACARTHUR'S MODEL FOR
VARIOUS NUMBERS OF HYPOTHETICAL SPECIES, S' (From Lloyd and
Ghelardi, 1964)
s'
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
-------
9.2.10 Reproductive Build. Reproductive requirements of fishes coupled with
early life history strategies enable a diversification of the ways habitats
are used. Balon (1975, 1981) divided reproductive modes of fishes in order of
evolutionary trends. Species are divided into nonguarders (guild A), guarders
(guild B), and bearers (guild C). The increase in evolutionary sophistication
from guilds A to C, generally conforms to levels of increased diversification
and reduction in niche overlap in complex environments (Table 6). Guild
dynamics are determined by three metrics in this category. The destruction of
diverse habitats not only reduce utilization of these habitats for
reproduction by adults, but also destroys nursery habitats for larval and
juvenile phases.
9.2.11 Hetrlc 5. Proportion of Non-guarding Guild A.I and A.2. The
nonguarding guild includes mostly r-stragegists which provide little parental
investment into each egg, usually possess early reproduction, small body size,
many small offspring, single production, and exhibit a type III mortality
(MacArthur, 1957; MacArthur and Wilson, 1967). Balon (1975) described the
nonguarding guild as broadcast spawners, usually without much developmental
specialization, and although may construct some nests always abandons them
post-reproduction. These species are often "pioneer" species and frequently
are dominant only in stressed and dominant only in stressed areas which are
periodically disturbed.
9.2.12 Metric 6. Proportion of Guarding Guild B.I and B.2. The guarding
guild typically include k-strategists as defined by MarArthur (1957) and
MacArthur and Wilson (1967). This strategy favors slower development, greater
competitive ability, delayed reproduction, larger body size, repeated
reproduction, fewer larger progeny, and exhibits types I and II mortality.
The guarding guild (Balon, 1975) is a solely ethological aspects of guild with
profound ecomorphological consequences. Better protected from enemies,
guarded eggs need not be numerous to assure survival of the species. As a
consequence, eggs can be larger and result in more viable offspring with less
food specialization. Spawning sites with low oxygen content can be used
because the guarding parents clean the eggs and produce a flow of water around
them by fin-fanning and oral ventilation. Fishes that do not build
complicated structures, nests, but that deposit their eggs on top of a
selected object, are also included in this section. The evolutionary
progression has been from (1) an exclusively parental male, (2) shared
parental care by the male and female, to (3) a division of roles with the
female as the direct parent and the male as the guardian to (4) polygyny
(Barlow, 1974).
9.2.13 Metric 7. Proportion of Bearers Guild C.I and C.2. This group is
divided into external and internal brooders (Balon, 1975). External brooders
carry their developing eggs on the surface of their bodies or in externally
filled body cavities or special organs. These include transfer, forehead,
mouth, gill-chamber, skin and pouch brooders. Internal brooders have eggs
fertilized internally before they are expelled from the body cavity. Special
organs are developed to facilitate sperm transfer. Mating does not
necessarily coincide with fertilization. After fertilization eggs can be
expelled and incubated externally or retained in the body cavity of the
217
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TABLE 6. CLASSIFICATION OF REPRODUCTION STYLES FOR FISHES IN ORDER
OF EVOLUTIONARY TRENDS (FROM BALON, 1981)
Ethological Section
A. Nonguarders
Ecological Group
A.I. Open Substratum Spawners
Guild
Selected key features of early
ontogeny
A. 1.1 Pelagic spawners
(pelagophils)
A. 1.2 Rock and gravel spawners with
pelagic larvae (lithopelagophils)
A.1.3 Rock and gravel spawners with
benthic larvae (lithophils)
Numerous buoyant eggs, none or
poorly developed embryonic
respiratory organs, little pigment,
no photophobia
Adhesive chorion at first, some eggs
soon buoyant, after hatching free
embryos pelagic by positive buoyancy
or active movement, no photophobia,
limited embryonic respiratory
structures*
Early hatched embryo photophobic,
hide under stones, moderately
developed embryonic respiratory
structures, pigment appears late
A.1.4 Nonobligatory plant spawners
(phytolithophils)
A.1.5 Obligatory plant spawners
(phytophils)
A.1.6 Sand spawners
(psammophils)
Adhesive eggs on submerged items,
late hatching, cement glands in free
embryos, photophobic, moderately
develop respiratory structures
Adhesive egg envelope sticks to
submerged live or dead plants, late
hatching, cement glands, not
photophobic, extremely will
developed embryonic respiratory
structures
Adhesive eggs in running water on
sand or fine roots over sand, free
embryos without cement glands,
phototropic, freely developed
respiratory structures, large
pectorals, large neuromast rods
(cupulae)
*See the final amendment in Balon (1981), page 389.
218
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TABLE 6, CLASSIFICATION OF REPRODUCTION STYLES FOR FISHES IN ORDER
OF EVOLUTIONARY TRENDS (FROM BALON, 1981) (CONTINUED)
Ethological Section
A. Nonguarders
Ecological Group
A.2. Brood hiders
Guild
Selected key features of early
ontogeny
A. 1.7 Terrestrial spawners
(aerophils)
A.2.1 Beach spawners
(aeropsammophils)
A.2.2 Annual fishes
(xerophils)
A.2.3 Rock and gravel spawners
(lithophils)
A.2.4 Cave spawners
(speleophils)
A.2.5 Spawners in live invertebrates
(ostracophils)
Small adhesive eggs scattered out of
water in damp sod, not photophobic,
moderately developed respiratory
structures
Spawning above the water line of
high tides, zygotes in damp sand
hatch upon vibration of waves,
pelagic afterward
In cleavage phase blastomeres
disperse and rest in 1st facultative
diapause, two more resting
intervals obligate--eggs and embryos
capable of survival for many months
in dry mud
Zygotes buried in gravel depressions
called redds or in rock interstices,
large and dense yolk, extensive
respiratory plexuses for exogenous
and carotenoids for endogenous
respiration, early hatched free
embryos photophobic, large emerging
alevins
A few large adhesive eggs, most hide
in crevices, extensive embryonic
respiratory structures, large
emerging larvae
Zygotes deposited via female's
ovipositor in body cavities of
mussels, crabs, ascidians or
sponges(?), large dense yolk, lobes
or spines and photophobia to prevent
expulsion of free embryos, large
embryonic respiratory plexuses and
carotenoids, probable biochemical
mechanism for immunosuppression
219
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TABLE 6. CLASSIFICATION OF REPRODUCTION STYLES FOR FISHES IN ORDER
OF EVOLUTIONARY TRENDS (FROM BALON, 1981) (CONTINUED)
Ethologlcal Section
A. Nonguarders
Ecological Group
A.2. Brood hiders
Guild
Selected key features of early
ontogeny
B.I.I Pelagic spawners
(pelagophils)
B.I.2 Above water spawners
(aerophils)
B.I.3 Rock spawners
(lithophils)
B.I.4 Plant spawners
(phytophi Is)
Nonadhesive, positively buoyant
eggs, guarded at the surface of
hypoxic waters, extensive embryonic
respiratory structures
Adhesive eggs, embryos with cement
glands, male in water splashes the
clutch periodically
Strongly adhesive eggs, oval or
cylindrical, attached at one pole by
fibers in clusters, most have
pelagic free embryos and larvae
Adhesive eggs each to variety of
aquatic plants, free embryos without
cement glands swim instantly after
prolonged embryonic period
Ethological Section
B. Guarders
Ecological Group
B.2 Nest spawners
Guild
Selected key features of early
ontogeny
B.2.1 Froth nesters
(aphrophils)
B.2.2 Miscellaneous substrate
and material nester
(polyphils)
Eggs deposited in a cluster of
mucous bubbles, embryos with cement
glands and well developed
respiratory structures
Adhesive eggs attached singly or in
clusters on any available
substratum, dense yolk with high
carotenoid contents, embryonic
respiratory structures well
developed, feeding of young on
parental mucus common
220
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TABLE 6. CLASSIFICATION OF REPRODUCTION STYLES FOR FISHES IN ORDER
OF EVOLUTIONARY TRENDS (FROM BALON, 1981) (CONTINUED)
Ethological Section
A. Nonguarders
Ecological Group
B.2. Nest spawners
Guild
Selected key features of early
ontogeny
B.2.3 Rock and gravel nesters
(lithophils)
B.2.4 Gluemaking nesters
(ariadnophils)
B.2.5 Plant material nesters
(phytophils)
B.2.6 Sand nesters
(psammophils)
B.2.7 Hole nesters
(speleophils)
B.2.8 Anemone nesters
(actiniariophils)
Eggs in spherical or elliptical
envelopes always adhesive, free
embryos photophobia or with cement
glands swing tail-up in respiratory
motions, moderate to well developed
embryonic respiratory structures,
many young feed first on the mucus
of parents
Male guards intensively eggs
deposited in nest bind together by a
viscid thread spinned from a kidney
secretion, eggs and embryos
ventilated by male in spite of well
developed respiratory structure
Adhesive eggs attached to plants,
free embryos hang on plants by
cement glands, respiratory
structures well developed in embryos
assisted by fanning parents
Thick adhesive chorion with sand
grains gradually washed off or
bouncing buoyant eggs, free embryo
leans on large pectorals, embryonic
respiratory structures feebly
developed
At least two modes prevail in this
guild: cavity roof top nesters have
moderately developed embryonic
respiratory structures. While
bottom burrow nesters have such
structures developed strongly
Adhesive eggs in cluster guarded
the base of sea anemone, parent
coats the eggs with mucus against
nematocysts, free embryo
phototropic, planktonic, early
juveniles select host anemone
at
221
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TABLE 6, CLASSIFICATION OF REPRODUCTION STYLES FOR FISHES IN ORDER
OF EVOLUTIONARY TRENDS (FROM BALON, 1981) (CONTINUED)
Ethological Section
B. Bearers
Ecological Group
C.I External bearers
Guild
Selected key features of early
ontogeny
C.I.I Transfer brooders
C.I.2 Auxiliary brooders
C.I.3 Mouth brooders
C.I.4 Gill-chamber brooders
C.I.5 Pouch brooders
Eggs carried for some time before
deposition: in cupped pelvic fins,
in a cluster hanging from genital
pore, inside the body cavity
(earlier ovoviviparous), after
deposition most similar to
nonguarding phytophils (A.1.4)
Adhesive eggs carried in clusters or
balls on the spongy skin of ventrum,
back, under pectoral fins or on a
hook in the superoccipital region,
or encircled within coils of
female's body, embryonic respiratory
circulation and pigments well
developed
Eggs incubated in buccal cavity
after internal synchronous or
asynchronous, or buccal
fertilization assisted by egg
dummies, large spherical or oval
eggs with dense yolk are rotated
(churning) in the cavity or densely
packed when well developed embryonic
respiratory structures had to be
assisted by endogenous oxydative
metabolism of carotenoids, large
young released
Eggs of North American cavefishes
are incubated in gill cavities
Eggs incubated in an external
marsupium: an enlarged and everted
lower lip, fin pouch, or membraneous
or bony plate covered ventral pouch,
well developed embryonic respiratory
structures and pigments, low number
of zygotes
222
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TABLE 6. CLASSIFICATION OF REPRODUCTION STYLES FOR FISHES IN ORDER
OF EVOLUTIONARY TRENDS (FROM BALON, 1981) (CONTINUED)
Ethological Section
B. Bearers
Ecological Group
C.2 Internal bearers
Guild
Selected key features of early
ontogeny
C.2.1 Facultative internal bearers
C.2.2 Obligate lecithotrophic
livebearers
C.2.3 Matrotrophous oophages and
adelphophages
Eggs are sometimes fertilized
internally by accident via close
apposition of gonopores in normally
oviparous fishes, and may be
retained within the female's
reproductive system to complete some
of the early stages of embryonic
development, rarely beyond the
cleavage phase: weight decreases
during embryonic development
(examples**: Galeus polli, Rivulus
marmoratus, Oryzias latipes)
Eggs fertilized internally, incubate
in the reproductive system of female
until the end of embryonic phase or
beyond, no maternal-embryonic
nutrient transfer: as in oviparous
fishes yolk is the sole source of
nourishment and most of the
respiratory needs; some
specialization for intrauterine
respiration, excretion and
osmoregulation: decrease in weight
during embryonic development
(examples: Torpedo oceJJata,
Poeciliopsis monadia, Poecilia
reticulata, Xenopoecilus poptae,
Schastes marinus)
Of many eggs released from an ovary
only one or at most a few embryos
develop into alevins and juveniles*,
feeding on other less developed
yoked ova present and/or
periodically ovulated (oophagy), and
in more specialized forms, preying
Note differences in the earier paper (Balon, 1975)
+Terminology as in Balon (1981).
223
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TABLE 6. CLASSIFICATION OF REPRODUCTION STYLES FOR FISHES IN ORDER
OF EVOLUTIONARY TRENDS (FROM BALON, 1981) (CONTINUED)
Ethologlcal Section
B. Bearers
Ecological Group
C.2 Internal bearers
Guild
Selected key features of early
ontogeny
C.2.3 Matrotrophous oophages and
adelphopages (continued)
on less developed sibling embryos
(adelphophagy): specialization for
intrauterine respiration, secretion
and osmoregulation similar to the
previous guild: large gain in weight
during intrauterine development
(examples: Lumma cornubica,
Eugamphodus temus, Latimeria
chalumnael)
C.2.4 Viviparous trophoderms
Internally fertilized eggs develop
into embryos, alevins or juveniles
whose partial or entire nutrition
and gaseous exchange is supplied by
the mother via secretory
histotrophes ingested or absorbed by
the fetus via epithelial absorptive
structures (placental analogues) or
a yolk sac placenta: small to
moderate gain in weight during
embryonic development (examples:
Galeus cam's, Myliobatis bovina,
MusteTus cam's, Sphyrna tiburo,
Zoarces viviparus, Ameca
splendens, Poeciliopsis turneri,
Heterandria formosa, Anableps dowi,
Embiotoca lateral is, Clinus
superciliosus)
224
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female, after which full-grown juveniles are born (Hoar, 1969; Balon, 1975,
1981).
9.2.14 Metric 8. Proportion of Simple Lithophil Mode of Reproduction. This
metric is used by Ohio EPA (1987) as a substitute in the adult IBI for
hybrids. Simple lithophils spawn where their eggs can develop in the
interstices of sand, gravel, and cobble substrates without parental care.
Generally, as the level of environmental degradation of simple lithophils
decreases. This is important in determining impacts from chronic levels of
exposure in sediments, and settling out of toxins in pools or backwater
habitats.
9.2.14.1 Abundance, Generation Time, and Deformity. Impacts to individuals
often are a compounding problem effecting community analyses. Reduction in
numbers of individuals, lowering of community mean generation time, and
increases in observed deformity and teratogenicity correspond with
environmental degradation. Loss of longer-lived species which require
specialized habitats (e.g., Acipenser fulvescens and Atractosteus spatula)
during reproduction and nursery are increasing at an alarming rate. Mean
generation time is a function of the time to first reproduction. This metric
may need further research before it can be utilized since it is proposed as a
community metric rather than as an individual metric as it was conceived.
9.2.15 Metric 9. Catch per Unit Effort. Population abundance varies with
ecoregion, stream size, and gear type used. It may be expressed as the catch
per unit effort, either by area, distance, or time sampled. Sites with lower
biological integrity will have reduced numbers of individuals, however,
rapidly flowing riffles should be excluded from comparison with pools and run
habitats (see spatial considerations). Organic enrichment usually increases
the number of individuals. Steedman (1988) addressed this situation by
scoring catch per minute of sampling. Unusually low numbers generally
indicate toxicity which is readily apparent at low levels of biological
integrity.
9.2.16 Metric 10. Mean Generation Time. Mean generation time is the average
age of parenthood, or the average age at which all offspring are born. A
longer-lived k-strategists species often spend several years before reaching
reproductive maturity, e.g., Salmonidae, Polyodontidae and Acipenseridae.
Vulnerability of these organisms to perturbations may have significant impact
to future recruitment during the larval and juvenile stages of development.
Mean generation time is an average value for a family based on life strategy
of representative taxa. Mean generation time is calculated as:
f = (a + w)/2
where:
a = age at first reproduction
w = age at last reproduction
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9.2.16.1 The community mean generation time is the sum of all generation
times for all families collected, divided by the total number of families.
9.2.17 Metric 11. Proportion of Deformity or Teratogenicity. lexicological
literature suggests that increased exposure to metals and organic chemical
compounds increases the proportion of teratogenicity among fathead minnows
(Birge et al., 1985; Simon, 1988). Additional effects have been documented in
a recent literature review by Weis and Weis (1989), as well as, exposure to
radiation (Lanthrop, personal communication). Teratogenic effects include
edematous yolk sacs, post caudal swellings, clear blood, reduced heart beat,
lack of fusiform shape, enlarged craniums, square eyes, or improper
development of the mandible (Simon, 1988). An increase in deformities or
teratogenicity is a result of increased exposure to toxic chemicals or
radiation. In reference and complex effluent testing using the fathead minnow
embryo-larval survival and teratogenicity test, one very infrequently observed
any teratogenicity in control samples. When deformities were observed they
were always less than 1% (Simon, personal communication).
9.2.17.1 Improperly preserved specimens will exhibit signs of deformity.
Birchfield (1987) determined that cranial anomalies were induced in
centrarchids and clupeids by fixing them in low concentrations of formalin
(<105), exposing them to high temperatures, or vigorously shaking the fixed
specimens. No cranial anomalies were found in larval fish fixed in formalin
solutions greater than 10% or in Bouin's fluid.
9.3 Taxonomic Considerations
9.3.1 The ability to differentiate families or larval fishes requires a basic
understanding of the morphometric and meristic characteristics which are
included in most taxonomic studies (Figures 1 and 2). Extensive literature
exists on specific families of larval or larval fishes and alternative
measurements, but certain standard measurements and counts continue to be the
main ones reported in the literature. The following explanation of how to
construct the character in question and the appropriate position to measure or
count the character is defined by Simon (1987) and Simon et al. (1987).
9.3.2 Characteristics are subdivided into morphometric, measurable
structures, and meristic, countable structures. Standard length and total
length are measured from the tip of the snout to the posterior portion of the
notochord and to the tip of the caudal finfold, respectively. Morphometric
measurements include head length—from the snout to pectoral fin origin; snout
length—from tip of the snout to anterior margin of eye; eye diameter--
anterior to posterior margin; preanal length--snout to posterior margin of
anus; body depth--vertical distance at anus; greatest body depth (also
referred to as shoulder depth or head depthj — largest vertical distance
(usually anterior dorsal finfold) or measured at origin of pectoral fin;
mid-postanal depth—vertical distance measured from dorsal to ventral margin
of body at anterior apex of the mean of the postanal myomeres; caudal peduncle
depth—vertical distance at anterior apex of penultimate myomere; head width--
measured dorsally at the posterior margin of eyes; yolk sac length and depth--
measured horizontally and vertically, respectively at the greatest distance on
the yolk sac.
226
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AS Anterior Margin of Snout
AE Anterior Margin of Eye
PE Posterior Margin of Eye
OP1 Origin of Pectoral Fin
(Head Length)
OD1 Origin of Spinous dorsal Fin
PV Posterior Margin of Vent
(Preanal Length)
OD2 Origin of Soft Dorsal Fin
MPM Mid-Postanal Myomere
AMPM Anterior Margin Penultimate
Myosepta
SL Standard Length
PC Posterior Margin of Caudal Fin
(Total Length)
Figure 1. Morphometric characteristics for larval fishes. The yolk sac (Y)
is included in width and depth measurements, but fin folds are not. "B" means
immediately behind, but not including, the eye or vent. Location of width and
depth measures at OD can only be approximated before the dorsal fin begins to
form. Fin length is measured along the plane of the fin from the origin to
the most distal margin. From Simon et al. (1987).
Total Length
Eye
Diameter
Caudal Peduncle
Length
Figure 2. Diagrammatic representation of morphology of a teleost larva,
From Auer (1982).
227
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9.3.3 Meristic measurements include the enumeration of all fin rays following
methods in Hubbs and Lagler (1958); head canal pores (Hubbs and Cannon, 1935);
preanal myomeres--those anterior to a vertical line drawn from the posterior
portion of the anus, including those bisected by the line, while postanal
myomeres include an urostylar element,
9.4 Provisional Key to the Families of North American Freshwater Fishes
(Adequate information is not available for all early life phases. Families
omitted from this key include Amblyopsidae, Cichlidae, Cyprinodontidae,
Poeciliidae, Umbridae, Cobitidae, Claridae, Oryziatidaw, and Elassomatidae).
Also see Section 12, Fisheries Bibliography, Subsection 12.5.2, Larval and
Immature Fishes.
KEY TO THE FAMILIES OF NORTH AMERICAN FRESHWATER FISHES
la. Body tubul ar, elongate, eel -1 i ke 2
Ib. Body not eel-like; usually with a single gill opening; stomodeum or
functional jaws present 3
2a. Body tubular, elongate, eel-like; seven gill openings; oral sucking disc
without jaws; lacking paired fins and distinct eyes Petromyzontidae
2b. Body eel-like; usually with a single gill opening; stomodeum, or
functional jaws present; eye large; processing paired fins...Anguillidae
3a. Barbels present on chin; mandibular barbels at corners of mouth; usually
hatching with some incipient fin rays present; yolk large usually with
complex vitell in veins Icataluridae
3b. Chin barbels and mandibular barbels absent; if barbels are present
limited to ventral portion of snout or single on chin....... ...4
4a. Adhesive disc present on snout; caudal fin heterocercal ....5
4b. Adhesive disc absent on snout.. 6
5a. Adhesive disc papillose; preanal myomeres number x; snout elongate with
remnant of adhesive disc until 20 mm total length (TL); dorsal and anal
finfolds originating posteriorly, finfold with dark triangular areas
near future dorsal, anal, and caudal Fins Lepisosteidae
5b. Adhesive disc smooth; preanal myomeres number x; without elongate snout,
dorsal finfold originating anterior pectoral fin; gular plate present;
body robust Amiidae
228
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6a. Larval 10-11 mm TL at hatching; preanal length 60-65% TL; yolk sac
large, oval, vascularized; barbels developing on ventral extension of
snout; head smal 1 7
6b. Larvae < 10 mm TL at hatching; preanal length greater than 60-65% TL;
large, oil globule; without barbels on ventral surface of snout 8
7a. Decreasing preanal length at increasing length, 65% TL becomes 60% TL >
11 mm; moderate dorsal finfold originates immediately behind head;
dorsal finfold origin length 25% TL; late protolarvae with four barbels;
dorsal fin origin posterior to vent; posterior margin of operculum not
extending past base of pectoral fin; scutes developing at juvenile
stages Acipenseridae
7b. Decreasing preanal length at increasing length, 60% TL becomes 50% TL at
> 11 mm; dorsal finfold originates at mid-preanal; dorsal finfold origin
length 35% TL; late protolarvae with two barbels; dorsal fin origin
anterior anus; posterior margin of operculum extending past base or
pectoral fin; no scutes developing at juvenile stages Polyodontidae
8a. Preanal length greater than 65% TL... 9
8b. Preanal length 60% TL or less 19
9a. Preanal length greater than 75% TL. ..10
9b. Preanal length between 65-75% TL . 13
lOa. Preanal length 76-89% TL; total myomeres greater than 45 12
lOb. Preanal length usually less than 75-79% TL; total myomeres less than 45
11
lla. Preanal myomeres > 27; mouth subterminal; body elongate, with usually
one to several rows of dorsal pigment.... Catosomidae
lib. Preanal myomeres > ; mouth superior, body elongate usually without
pigmentation dorsally Clupeidae
12a. Postanal myomeres 13-17; yolk sac small, round and far forward......
Osmeridae
12b. Postanal myomeres < 10; yolk sac larger, elongate or oval, situated
posteriorly. Clupeidae
13a. Preanal myomeres greater than or equal to 40 14
13b. Preanal myomeres less than 40 ....15
229
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14a. Postanal myomeres 14015; preanal length 72-75% TL; adipose fin present;
swim bladder visibly present Osmeridae
14b. Postanal myomeres 15-22; preanal length 67-72% TL; adipose fin absent;
swim bladder not visible Esocidae
15a. Yolk sac long, bilobed with the anterior portion thick and oval,
posterior section thick and tubular, preanal length 58-74% TL 16
15b. Yolk sac not bilobed, either elongate or oval; if bilobed usually with
both sections of equal portion; preanal length 68-75% TL 17
16a. Larvae densely pigmented, evenly over body, with a dark stripe over
gut; usually less than 27 preanal myomeres; body robust Cyprinidae
16b. Pigmentation limited to dorsum, usually on cranium and sometimes
mid-dorsally in two to four distinct rows; body elongate Catostomidae
17a. Preanal myomeres < 31, postanal myomeres less than 41 Catostomidae
17b. Preanal myomeres > 31 18
18a. Postanal myomeres < 41; larvae large, at 7 mm still possess yolk;
preanal length 62-64% TL Hiodontidae
18b. Postanal myomeres > 41; preanal length 67-74% TL.. Cyprinidae
19a. Preanal length > 48% TL 20
19b. Preanal length < 48% TL 27
20a. Preanal > 56% TL 21
20b. Preanal 48-15% TL 23
21a. Preanal myomeres < 26; preanal length 56-58% TL; larvae large, yolk sac
present until 7-10mm TL. Hiodontidae
21b. Preanal myomeres < 26; preanal length < 56% TL; yolk sac larvae < 7 mm
TL 22
22a. Preanal myomeres 8-12; postanal myomeres 9-15 Moronidae
22b, Preanal myomeres 15-26; postanal myomeres 18-26 Percidae
23a. Preanal myomeres > 15 Percidae
23b. Preanal myomeres < 15 , 24
24a. Total myomeres < 26 Moronidae
24b. Total myomeres > 26 25
230
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25a. Preanal myomeres 14-16; preanal length > 50% TL Gasterosteidae
25b. Preanal myomeres < 26
26a. Postanal myomeres < 19; gut massive, uncoiled; pectoral fins
proportional Centrarchidae
26b. Postanal myomeres > 19; large pectoral fins 27
27a. Preanal length < 35%; preanal myomeres 6-7; postanal
myomeres 28-31 Atherinidae
27b. Preanal length > 35% , 28
28a. Postanal myomeres approximately 40; preanal length 39-44% TL Gadidae
28b. Postanal myomeres much less than 40; preanal length 44% TL 29
29a. Postanal myomeres < 11; posterior oil globule in yolk sac 30
29b. Postanal myomeres > 20; mouth terminal to superior;
preanal length 45% TL., 30
30a. Postanal myomeres > 30; mouth terminal to superior; preanal
length 45% TL.. .Fundulidae
30b. Postanal myomeres < 20; mouth subterminal to inferior; preanal
length 45% TL Percopsidae
9.5 Fish Larvae Sampling Precision
9.5.1 When investigators collect larval fish samples, the accuracy of the
sampling methods and equipment must be carefully considered. Using literature
data, Cyr et al. (1992) demonstrated that past sampling designs have been
inadequate for the comparison of larval fish abundance across sites or time
periods. Therefore, Cyr et al. (1992) developed a general model based on
published data to predict the variance in larval fish abundance among
replicate samples and provided guidelines for estimating the number of larval
fish samples necessary to obtain acceptable or desired levels of precision at
a collecting site. For studies that include large aquatic habitats of many
sites as well as changes in abundance through time, they concluded that
investigators must consider patterns of spatial and temporal variation when
sampling larval fish populations. They also indicated that in arriving at an
efficient allocation of sampling effort, that each scale of variation must be
considered. Furthermore, careful consideration of precision in the context of
data quality objectives (DQOs) (See Section 2, Quality Assurance and Quality
Control) will improve the qualitative or quantitative evaluations of
ichthyoplanktonic population studies.
231
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9.6 Literature Cited
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fishes in low-velocity riverine habitats. Prog. Fish-Cult. 46:59-62.
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promelas) subchronic toxicity test. Env. Tox. Chem. 4:711-718.
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of main channel ichthyoplankton abundance. Ohio river Miles 569 to 572.
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comparative descriptions of freshwater and marine larval fishes. Early
Life History Section Newsletter 7:10-15.
Simon, T.P. 1987. Description of eggs, larvae and early juveniles of the
stripetai1 darter, Etheostoma kennicotti (Putnam) and spottail darter, f.
236
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squamiceps Jordan (Percidae: Etheostomatini) from tributaries of the Ohio
River. Copeia 1987:433-442.
Simon, T.P. 1988. Subchronic toxicity evaluation of the grand calumet River
and Indiana Harbor Canal using the embryo-larval survival and
teratogenicity test. Proc. Ind. Acad. Sci. In Press.
Simon, T.P. 1989. Rationale for a family-level ichthyoplankton index for
use in evaluating water quality. In: W.S. Davis and T.P. Simon (eds.).
EPA-905/9-89/007. Proceedings of the 1989 Midwest Pollution Control
Biologists Meeting, Chicago, IL. pp. 41-65.
Simon, T.P. 1990. Predictive abilities of environmental Protection Agency
subchronic toxicity endpoints for complex effluents. Proc. Ind. Acad.
Sci. 99:29-37.
Simon, T.P. and R. Wallus. 1989. Contributions to the early life history of
gar (Actinopterygii:Lepisosteiformes) from the Ohio and Tennessee River
basins with emphasis on larval taxonomy. Trans. KY Acad. Sci. 50:59-74.
Simon, T.P., R. Wallus, and K.D. Floyd. 1987. Descriptions of protolarvae of
seven species of the darter subgenus Nothonotus with comments on
intrasubgeneric characteristics. Am. Fish Soc. Symposium 2:179-190.
Synder, D.E. 1981. Contributions to a guide to the cypriniform fish larvae
of the upper Colorado River system in Colorado. U.S. Bur. Land Manag.,
Denver, CO.
Snyder, D.E. 1983. Fish eggs and larvae. In: L.A.Nielsen and D.L. Johnson
(eds.). Fisheries Techniques. Am. Fish. Soc., Bethesda, MD. pp.165-
198.
Steedman, R.J. 1988. Modification and assessment of an index of biotic
integrity to quantify stream quality in southern Ontario. Can. J. Fish.
Aquat. Sci. 45:492-501.
Sturm, E.A, 1988. Descriptions and identification of larval fishes in
Alaskan freshwaters. M.S. Thesis, Univ. Alaska, Fairbanks, Alaska.
Taber, C.A. 1969. The distribution and identification of larval fishes in
the Buncambe Creek arm of Lake Texoma with observations on spawning
habits and relative abundance, Ph.D. Dissertation, Univ. OK, Norman, OK.
Tuberville, J.D. 1979. Drift net assembly for use in shallow water. Prog.
Fish-cult. 41:96.
USEPA. 1973. Biological field and laboratory methods for measuring the
quality of surface waters and effluents. C.I. Weber (ed.). EPA-670/4-
73/001. U.S. Environmental Protection Agency, Office of Research and
Development, Cincinnati, OH.
237
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USEPA, 1990. Macroinvertebrate field and laboratory methods for evaluating
the biological integrity of surface waters. Donald J. Klemm, Philip A.
Lewis, Florence Fulk, and James M. Lazorchak. EPA/600/4-90/030.
Environmental Monitoring Systems Laboratory, U.S. Environmental
Protection Agency, Cincinnati, OH
Wallus, R, 1986. Paddlefish reproduction in the Cumberland and Tennessee
River systems. Trans, am. Fish. Soc. 115:424-428.
Wallus, R. and J.P. Buchanan. 1989. Contributions to the reproductive
biology and early life ecology of mooneye in the Tennessee and Cumberland
Rivers. Am. Midi. Nat. 112(1):204-207.
Wallus, R., T.P. Simon, and B.L. Yeager. 1989. Contributions to the
reproductive biology and early life histories of Ohio River basin fishes.
Vol. I. Acipenseridae to Clupeidae. Tennessee Valley Authority,
Knoxville, TN.
Wang, J.C.S. 1981. Taxonomy of the early life history stages of fishes-
fishes of the Sacramento-San Joaquin Estuary and Moss Landing Harbor-
Elkhorn Slough. California. EA Publication, Concord, CA.
Wang, J.C.S. and R.J. Kernehan (eds.). 1979. Fishes of the Deleware
estuaries: A guide to the early life histories. EA Publications, Towson,
MD.
Washington, H.G. 1984. Diversity, biotic and similarity indices, a review
with special relevance to aquatic ecosystems. Water Res. 18:653-694.
Weis, J.S. and P. Weis. 1989. Effects of environmental pollution on early
fish development. Reviews Aquatic Sci. 1:45-73.
238
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SECTION 10
FISH HEALTH AND CONDITION ASSESSMENT METHODS1
10.1 Introduction
10.1.1 The fish health and condition assessment methods provide relatively
simple and rapid indication of how well fish live in their environment. They
are manifestations of biochemical and physiological alterations expressed at
the organism level. Goede and Barton (1990) and Goede (1992) review various
types of condition indices that can be used to assess stress in fish, and they
also describe an empirical necropsy-based system of organ and tissue indices
that provides a fish health and condition profile of fish populations.
External aspects, blood parameters, and the normal appearances of internal
vital organs are assumed to indicate that a fish population is in harmony with
its environment, or if the fish have been challenged, that the animals have
not been stressed enough to cause obvious structural changes. When the
necropsy system is applied in the field, departure from normal growth,
bioenergetic state, and general homeostasis can be detected, as well as the
presence of infectious agents in fish. Advantages of these methods over
physiological monitoring or community analyses are that they are simple to
use, requires little training, and does not need costly, sophisticated
equipment. The fish health and condition assessment could be used routinely
in research, culture, management, and regulatory programs to establish a data
base for evaluating whether a fish population is coping successfully with its
environment.
10.1.2 Novotny and Beeman (1990) evaluated the fish health and condition
assessment methods on juvenile chinook salmon (Oncorhynchus tshawytscha) that
were reared in net pens in the Columbia River, Washington, and they found the
procedures were efficient in assessing the condition of fish held under
various rearing conditions. They, furthermore, concluded that the simplicity
of the methods makes them useful for monitoring fish in culture facilities and
fish from wild stocks. These methods are meant to be used by investigators
who routinely work in the field and for determining the general health and
condition of a group of fish.
10.1.3 It is important that the investigator be able to use the minimum of
equipment needed for these methods and to be able to recognize gross
appearance or differences of systems in tissues and organs. The investigator
does not specifically have to be able to diagnose the cause or causes of the
condition. If a departure from normal condition is evident in a significant
proportion of the fish population, it is appropriate that a specialist be
called to help determine the cause of the variation.
10.1.4 A list of equipment and materials for the fish health and condition
assessment is found in Table 1.
Adapted from Goede and Barton (1990) and Goede (1992).
239
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TABLE 1. EQUIPMENT AND MATERIALS FOR FISH HEALTH AND CONDITION ASSESSMENT
- Microhematocrit Centrifuge
- Microhematocrit tubesa'b
- Critoseal clay to seal hematocrit tubes
- Microhematocrit tube reader
- 1.0 percent sodium or ammonia heparin solution
- Hand held serum protein refractometer
- Lens paper
- Bunsen Burner to sharpen hematocrit tubes
- Sharp/blunt scissors
- Dissecting forceps (preferably a small "mouse tooth type")
- MS-222 or comparable anesthetic6
- Metric scale to weigh individual fish
- Fish measuring board
- Hand held magnifying glasses for small fish
- Buckets and tubs to handle fish
- Calculators with standard deviation button
Heart puncture:
8Using capillary tubes: Sharpen capillary tubes and re-heparinize sharpened
end at least 1/3 to 1/2 of tube.
bHeparin:
Use 0.1 gm of heparin to 10 mL distilled water. Fill capillary tube 1/3 to
1/2, then drain back into heparin solution. This solution can be reused
again for rest of tubes. Remove all heparin from tubes and dry tubes
overnight.
cMS-222 Mixture:
To incapacitate but not kill. A solution in excess of 50 mg/L (ppm) MS-222 is
recommended. Use 4 times this amount for lethal dosage.
240
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10.2 Sampling and Collection of Fish
10,2.1 The desired sample size for this procedure is 20 fish of the same
species. When working with free-ranging populations, it is not always easy to
obtain fish. In the field, the samples often are collected from fish captured
in routine netting or electrofishing operations. In some sampling situations
20 fish of the same species might be difficult to collect. In this
circumstance the investigator must work with what is caught.
10.2.2 The composition of the fish sampled (e.g., age class, length grouping,
etc.) depends upon the data quality objectives (DQOs) of the investigation and
upon what fish are available (see Section 2, Quality Assurance and Quality
Control).
10.3 Handling of Fish
10.3.1 The ideal collection is taken alive and handled carefully until they
can be anesthetized. The fish should be immobilized shortly after capture
with an appropriate anesthetic, e.g., tricaine MS-222 (see Table 1).
10.4 Sampling and Reading of Blood
10.4.1 Blood should be collected by cardiac puncture with a sharpened,
heparinized microhematocrit tube. If blood is needed for purposes in
addition to those of this procedure, a larger volume can be sampled with a
syringe and needle from the caudal vasculature. The microhematocrit tube can
then be filled from that volume with the syringe. The tube, once filled, is
plugged on one end using a commercial clay, prepared and sold for that
purpose. It is advised that you place the filled tubes upright in a rack with
numbered holes to await placement into a centrifuge. Every effort should be
made to keep the tubes in order so that they can be accurately matched to the
fish from which they were taken. The tubes are then placed in the numbered
slots of a microhematocrit centrifuge and spun for five minutes. A typical
microhematocrit centrifuge develops approximately 13,000 G. Erythrocytes (red
blood cells) have been shown to "swell" when exposed to carbon dioxide. Thus,
it is important that the tubes be spun within one hour of sampling. Once the
tubes have been centrifuged they can be transported and read in a more
convenient location but they should be read within two hours and definitely
before the plasma begins to coagulate. Once the blood fractions have been
separated by centrifuging, you can remove the tubes and place them again in
the numbered rack. Always keep them in the order in which they were collected
so they can be matched with the individual fish from which they were
collected. The tubes can be kept until later or one can proceed to read the
hematocrit, leucocrit, and plasma protein.
10.4.2 Hematocrit is the packed red cell volume of the blood and is expressed
as a percentage of the total column. It is obtained by placing the
centrifuged tubes on a microhematocrit reader. These are available in several
styles and costs but the simple plastic reader cards containing a nomograph
are preferred. The tube is placed on the card so that the bottom of the red
(erythrocytes) portion of the column is at the zero line and the meniscus of
the clear plasma portion of the column is on one hundred percent. The
241
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location of the top of the red portion indicates the volume percentage of red
blood cells or hematocrit.
10.4.3 There is usually a small "buffy or gray" zone just above the red zone.
This is composed of the leucocytes or white blood cells and is used to
estimate the leucocrit or percent leucocytes in the packed column. The card
reader can be used to read this, and a small magnifying glass is helpful.
10.4.4 Next, the protein content of the plasma is determined. This is done
by carefully breaking the hematocrit tube just above the "buffy" zone to
obtain only the clear plasma fraction. Be sure that there are no small glass
fragments on the broken end and then express the clear plasma onto the glass
surface of the hand-held protein refractometer. Read the weight/volume
percent of protein. The refractometer must be calibrated before use. To do
this, place a few drops of distilled water on the prism surface and adjust the
boundary line to the "w" or "wt" mark with the adjusting screw. Some
instruments have a thumbscrew and some require a small screwdriver. The
investigator should consult the manual supplied with the unit in question.
The instrument should be cleaned between readings with lens paper to avoid
scratching the surface. The surface should be cleaned with water and dried
with lens paper after every use.
10.5 Length and Weight Heasurements
10.5.1 The lengths and weights can be measured immediately after the blood
samples have been collected for hematocrit determinations.
10.5.2 The total length of each fish should be determined in millimeters and
the weight in grams. This is fairly straight forward but might be pointed out
that the length and weight were initially included in the procedure to see if
there was any correlation between fish size and the other parameters.
10.5.3 If it is desired to obtain an accurate estimate of size of the fish in
the population, more lengths and weights should be taken through non-lethal
sampling. The computer program, discussed later, will accommodate 60 fish.
10.6 External Examination
10.8.1 When the fish (Figure 1. External features of a composite fish) are
laid out in front of you it is the best time to make general observations
about the fish. Record general remarks about fins, skin, and other external
features before you begin the specific observation of particular organs and
systems. Important conditions to note are deformities, scale loss, and
external parasites. These observations are carried as remarks in the data
base. It must be noted here that primary observations included in this
procedure were intended to permit some inference with respect to health and
condition of the fish. This is only one aspect of "quality". Observations
relative to esthetics are included as remarks only. Fish species (e.g.,
Catostomidae, Cyprinidae) develop cornified epithelial tubercles and engage in
nuptial bouts. If external lesions or scars are observed in some specimens,
the possibility of external anomalies related to spawning behavior should be
noted.
242
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10.6.2 Begin the observations as outlined in the classification system (Table
2), Be sure to record all observations using the abbreviations or codes
listed on the classification scheme. This is necessary for subsequent entry
into the computer program (see AUSUM PROGRAM USE, page 270). If the
observation does not seem to fit any of the listed categories, list it as OT
which indicates "other". If you use this category be sure to describe it in
the remarks column. It is much easier for the recorder if you proceed
routinely in the same order laid out on the fish necropsy (postmortem
examination) worksheet (Figure 2). There are many systematic approaches to
the order of the procedures, but Goede (1992) has found it more efficient to
"open" all of the fish first with the use of sharp/blunt scissors by making a
ventral cut from the anal vent forward to the pectoral girdle, cutting closely
to one side of the pelvic girdle. A short distance of the "hind gut" is
opened with this first cut to permit later observation. Do not insert the
scissors so far that the internal organs are damaged. The fish are opened and
laid down in front, in proper order, to wait the final inspection,
10.6.3 Take into consideration the circumstances of the collection. If the
fish were collected dead, you must be aware of the often subtle differences
this can make in appearance of organs and tissues while still permitting valid
observation within the context of this procedure. A photographic, colored
atlas (Goede, 1988) of necropsy classification categories has been prepared
and may be obtained from Ronald W. Goede, Utah Division of Wildlife Resources,
Fisheries Experiment Station, 1465 West 200 North, Logan, Ut. 84321-6233. The
cost of the atlas is $80.00.
10.7 External Organs
10.7.1 Eyes
10.7.1.1. Normal (N) - no aberrations in evidence. Good "clear" eyes.
10.7.1.2 Exopthalmia (El or E2) - Swollen, protruding eye. More commonly
referred to as "popeye". It is coded as El or E2. This refers to the
presence of exopthalmia in one eye or two eyes.
10.7.1.3 Hemorrhagic (HI or H2) - Refers to bleeding in the eye. "Blind" (Bl
or B2) - This is a very graphic category and you need not know whether the eye
is functionally blind. It generally refers to opaque eyes, and the opacity is
not important here.
10.7.1.4 "Missing" (Ml or M2) - An eye is actually missing from the fish.
10.7.1.5 "Other" (OT) - Any manifestations which do not "fit" the above.
Describe in the remarks column.
10.7.2 Gills
10.7,2.1 Normal (N) - no apparent aberrations in gills. Be very careful in
this observation. The gill can easily be effected by the manner in which the
fish is handled during and after collecting.
243
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FIN MEMBRANE SPINY-RAYED PORTION
OF DORSAL FIN
NAP,E ^^/Tht-rt^" SOFT-RAYED PORTION
OPERCLE
(GILL COVER)
LATERAL LINE SCALE
SNOUT PUPIL
BARBELS^
NARIS
(NOSTRIL) \jXfgj
UPPER
JAW
LOWER
JAW
$&«. OF DORSAL FIN
\&.-$5
"EAR FLAP"
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un BODY
GILL Yin—SCALES
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ISTHMUS W/??^)^
(THROAT) W
MAXILLARY \jg£y APPENDAGE
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PECTORAL
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ANAL FIN
CAUDAL FIN
(TAIL FIN )
PELVIC
FIN
Figure 1. External features of a composite fish. From Lagler (1962), Atlas of Fish
Anatomy, Plate 1, Michigan Fisheries No. 5, Department of Fisheries, School
of Natural Resources, The University of Michigan, Ann Arbor, MI.
-------
TABLE 2. NECROPSY CLASSIFICATION OUTLINE
Length:
Weight:
Ktl:
Eyes:
Gills:
Pseudobranch:
Thymus:
Fins:
Opercles:
Mesentery Fat;
Spleen:
Hind Gut:
Kidney:
Liver:
Total length in millimeters
Weight in grams
W x 10s
See Subsection 10.9.
Normal (N), Exopthalmia (El, E2), Hemorrhagic (HI, H2),
Blind (Bl, B2), Missing (Ml, M2), Other (OT)
Normal (N), Frayed (F), Clubbed (C), Marginate (M), Pale
(P), Other (OT)
Normal (N), Swollen (S), Lithic (L), Swollen and Lithic
(S&L), Inflamed (I), Other (OT)
No Hemorrhage (0), Mild Hemorrhage (1), Severe Hemorrhage (2)
No active erosion or previous erosion healed over (0), Mild
active erosion with no bleeding (1), Severe active erosion
with hemorrhage and/or secondary infection (2)
No shortening (0), Mild shortening (1), Severe shortening (2)
Internal body fat expressed with regard to amount present:
0 - None
1 - Little, where less than 50% of each cecum is covered
2 - 50% of each cecum is covered
3 - More than 50% of each cecum is covered
4 - Ceca are completely covered by large amount of fat
Black (B), Red (R), Granular (G), Nodular (NO), Enlarge (E),
Other (OT)
No inflammation (0), Mild inflammation (1), Severe
inflammation (2)
Normal (N), Swollen (S), Mottled (M), Granular (G),
Urolithic (U), Other (OT)
Red (A), Light red (B), "Fatty" liver, "Coffee with cream"
color (C), Nodules in liver (D)? Focal discoloration (E),
General discoloration (F), Other (OT)
245
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TABLE 2. NECROPSY CLASSIFICATION OUTLINE (CONTINUED)
Bile: 0 - Yellow or straw color, bladder empty or partially full
1 - Yellow or straw color, bladder full, distended
2 - Light green to "grass" green
3 - Dark green to dark blue-green
Blood: Hematocrit - Volume of red blood cell (erythrocytes)
expressed as percent of total blood volume.
"Buffy" zone of the packed cell column.
Leucocrit - Volume of white blood cells (leucocytes)
expressed as percent of total blood volume.
"Buffy" zone of the packed cell column.
Plasma Protein - Amount of protein plasma, expressed as gram
percent (grams per 100 mL).
246
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Fish Necropsies
Wildlife Resources
2/91 FES-25
PO
Dale Unit Strain Quality
Location Fish Source
Mark/Lot
Age
Control *
Ca
se History #
Hat. Date Tissue Collection *
liwestigatof(s) Water Temp.
Water Hardness
Reason lor Autopsy Remarks
Smp
no
1
2
3
4
5
6
7
e
a
10
n
12
13
14
15
16
17
ia
19
20
Lglti
mm
Watrt
gm
M
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Sex
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Pro
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Opt
Remarks
Flns_
SWn
GENERAL REMARKS
Qonads
Other
Figure 2. Fish necropsy worksheet.
-------
10.7.2.2 "Frayed" (F) - This generally refers to erosion of tips of gill
lamellae resulting in "ragged" appearing gills. Mere separation of gill
lamellae can be construed to be "frayed" but that condition may have been
caused by something as simple as the manner in which the gill was exposed by
the investigator.
10.7.2.3 "Clubbed" (C) - This refers to swelling of the tips of the gill
lamellae. They can often appear bulbous or "club-like". The causes are not
pertinent until interpretation is considered.
10.7.2.4 "Marginate" (M) - a graphic description of a gill with a light
discolored margin along the distal ends or tips of the lamellae or filaments.
Margination can be and often is associated with "clubbing". If both (C) and
(M) seem to apply, it is not a problem. It is important that you note that it
was not normal. Use the one which seems most appropriate.
10.7.2.5 "Pale" (P) - This refers to gills which are definitely very light in
color. Severe anemia can result in gills which are discolored to the point of
being white. Severe bleeding induced during sampling of blood can also result
in somewhat pale gills. Gills begin to pale somewhat after death also. This
is not uncommon in fish taken from nets. All of this should be considered in
making the observation.
10.7.2.6 Other (OT) - Any observation which does not fit above. Describe in
remarks.
10.7.3 Pseudobranchs (The pseudobranch is located dorsally and anterior to
the gills in the branchial cavity and can be easily observed under the
opercula.) Some species lack pseudobranchia entirely.
10.7.3.1 Normal (N) - The normal pseudobranch is quite "flat" or even concave
in aspect and displays no aberrations.
10.7.3.2 Swollen (S) - The "swollen" pseudobranch is convex in aspect and not
difficult to discern upon close examination.
10.7.3.3 Lithic (L) - Mineral deposits in pseudobranchs, manifested by
appearance of white, somewhat amorphous spots or foci.
10.7.3.4 Swollen and Lithic (S&L) - Lithic pseudobranchs are often also
swollen.
10.7.3.5 Inflamed (I) - This is a generic use of the term, inflamed, and
would more appropriately be termed "redness" because it also includes
observations of hemorrhage and any other cause of redness. The term,
"inflamed" has been traditionally used to describe this condition and is thus
contained for that reason.
10.7.3.6 Other (OT) - This term will cover any manifestation observed in the
pseudobranch which is not covered in the categories. Be sure to describe in
remarks.
248
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10.7.4 Thymus (Assessment of the thymus involves degree of petechial or
"pinpoint" hemorrhage).
10.7.4.1 No Hemorrhage (0) - The thymus displaying no hemorrhage is
considered to be a normal condition, although this assumption is still under
investigation. Caution must be exercised here because when the thymus
involutes or ceases to function there is no observable petechial hemorrhage.
This happens normally as the fish mature. In salmonids involution of the
thymus is thought to happen at two or three years of age but there is
considerable disagreement among investigators about this point.
10.7.4.2 Mild Hemorrhage (1) - A few red spots or petechial hemorrhages in
evidence. This might be only two or three small spots.
10.7.4.3 Severe Hemorrhage (2) - Many "pin point" hemorrhages in evidence
with some of them coalescing. The general area may also have a swollen
tumescent appearance but that should be recorded in remarks.
10.7.5 Fins - It must be remembered that this particular assessment procedure
is concerned primarily with health and condition. It is not concerned with
aesthetic values. Eroded or "ragged" fins are definitely indicative of a
departure from normal condition and health. Previously eroded fins which are
completely healed over and showing no evidence of the active erosion are, for
the purposes of this assessment, considered normal. The evaluation of fins is
relative to the degree of active erosion process in evidence. For the
purposes of this procedure the number and location of fins involved is not
significant. If only one fin is displaying active erosion, the observation
must be ranked and recorded. If several fins are displaying erosion with
unequal severity, the observation must refer to the most severe in evidence.
This unequal nature of the observations, in this case, is less significant in
a full 20 fish sample. The classification is as follows:
10,7.5.1 No Active Erosion (0) - Normal appearing fins with no active
erosion. This would include previously eroded fins which were completely
healed over.
10.7.5.2 Mild active erosion (1) - Active erosion process but no hemorrhage
or secondary infection in evidence.
10.7.5.3 Severe Active Erosion (2) - Active erosion with hemorrhage and/or
secondary infection in evidence.
Note: Make a general remark relative to which fins were involved and any
other observation of special significance. There is a space for this type of
entry at the bottom of the data collection worksheet. This is particularly
important in the summary.
10.7.6 Opercles (It is necessary only to observe the degree of shortening of
the opercles. The classification is as follows:)
10.7.6.1 Normal Opercle (0) - No shortening; gills completely covered.
249
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10.7.6.2 Slight Shortening (1) - Slight shortening of the opercle with a very
small portion of the gill exposed
10.7.6.3 Severe Shortening (2) - Severe shortening of the opercles with a
considerable portion of the gill exposed.
10.8 Internal Examination (or Necropsy)
10.8.1 Figure 3 reveals the key internal anatomical features of a typical
soft-rayed fish (brook trout), and Figure 4 displays the anatomical features
of a characteristic spiny-rayed fish (largemouth bass).
10.8.1.1 If the fish was not "opened" as suggested above, it should be done
now to permit access to the internal systems. Remember to proceed, where
possible, in the order listed on the data sheets. This facilitates recording.
The order was established beginning posteriorly with the mesenteric fat depot,
proceeding anteriorly through the spleen and hindgut, to the kidney, liver,
and gall bladder, to the gonads for determination of gender and state of
development. At this point, it is wise to observe the mesentery tissue for
hemorrhage or inflammation and record in remarks if not normal.
10.8.2 Mesenteric Fat
10.8.2.1 The ranking of mesenteric fat depot has been developed around
salmonid fishes with prominent pyloric caeca. It must be noted here that
there is great variation among the different fish species in the way that they
store this fat. If the system is to be applied to other groups of fishes,
alternate ranking criteria will have to be developed. It should be further
noted that as long as the ranking is 0 through 4 the computer program, AUSUM,
for summarizing data, can still be used. The following ranking system was
developed for the rainbow trout but has been applied with minor variations to
all major groups of salmonids.
0 - No fat deposited around the pyloric ceca. If there is no fat deposit in
evidence anywhere in the visceral cavity it is clearly a "0" fat.
1 - Slight, where less than 50% of each cecum is covered with fat. There are
cases where there will be no fat in evidence on the ceca, but there will be a
slight fat currently classes as a "1".
2 - 50% of each cecum is covered with fat.
3 - More than 50% of each cecum is covered with fat.
4 - Pyloric ceca are completely covered by a large amount of fat.
10.8.3 Spleen
Black (B) - The "black" is actually a very dark red color of the spleen.
Red (R) - Red coloration of the spleen. There is subjective variation among
250
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investigators as to whether the spleen is black or red, but both conditions
are considered normal
Granular (G) - Granular or "rough" appearance of the spleen.
Nodular (NO) - The spleen contains or manifests fistulas or nodules of varying
sizes. These are often cysts, such as those encountered with mycobacterial
infections.
Enlarged (E) - Spleens can, on occasion, be significantly and noticeably
enlarged.
Other (OT) - Occasionally there are observable, gross aberrations which do not
fit the above. There may be spleens with a gray mottling and some with very
small spleens. These should be classed as "other" and described in remarks.
10.8.4 Hindgut
10.8.4.1 A short distance of the hind gut should be "opened". This should,
in fact, have been accomplished as mentioned above when the body cavity is
incised. If not, it must be opened to expose the "inner lining" or mucosa.
Using the handle of a forceps or some other appropriate blunt instrument,
lightly "scrape" out the contents of the hindgut so that you can observe
relative reddening or inflammation.
No inflammation (0) - No inflammation or reddening of the hindgut.
Slight inflammation (1) - Mild or slight inflammation or reddening of the
hindgut.
Severe inflammation (2) - Considerable, severe inflammation or reddening of
the high gut.
10.8.5 Kidney
Normal (N) - Good firm dark red color lying relatively flat dorsally in the
visceral cavity along the length of the ventral surface of the vertebral
column. It will be necessary to pull the swimbladder and some of the
mesentery aside to expose the kidney to view.
Swollen (S) - Enlarged or swollen wholly or in part.
Mottled (M) - Gray discoloration, mottled or "patchy" in appearance ranging
from scattered patches of gray to total gray discoloration. This is not to be
mistaken with the superficial gray appearance induced by the mesenteric
membrane on the surface of the kidney. This should be moved aside before
observation is recorded.
Granular (G) - The kidney may have a "granular appearance and texture. This
may be induced by granulomatous concretions.
251
-------
SUPSIAOCCIPlTAL
CAUDAL PEDUNCLE
DERMilHMOlO
PREMAX!LLAH*\
r\>
in
PO ENTOCLOSS
/ / / PNEUMATIC
/ DUCT
PtCtQRAL
GIRDi-C
\
\INTISTINt
Pt'l V)C ClN
SCALE
2
3 IHCHO
Figure 3. Anatomy of a soft-rayed bony fish, the brook trout, 5a?veJ/m/s fontinalis. From Lagler
(1962), Atlas of Fish Anatomy, Plate IV, Michigan Fisheries No. 5, Department of Fisheries,
School of Natural Resources, The University of Michigan, Ann Arbor, MI.
-------
SPINOUS PORTION
OF DORSAL FIN
SOFT-RAYED PORTION
OF DORSAL FIN
DORSAL SPINE
INTERNEURAUS
SUPRAOCCIPITAL
FMSNTAL
DF.HME1HMOIO
VOMCH
OHA1N
PREMAKILLAHY
tn
w
iv\ \ \ \ \ - u^-
rflV^ -^^-<%«-
A *»*£&*'"''"P**1"
OENTAHt
ENTOOLOSSAL
BASIBRANCHIAl
CAUDAL FIN
.,.-..,.,»
VENTRAL / HEART
AORTA
BRANCHIOSIEGAL SAT PECTOHAL
6!RDLE
( INTESTINE
OVARY
PYIORIC \ \f,ffti
CAtCA \ bH-ttN
* URINARY
STOMACH BLADDER
ANAL SPINE
INTER HEMALS
INCH
PELVIC FIN
Figure 4. Anatomy of a spiny-rayed bony fish, the largemouth bass, W/cropterus sa/mo/des. From Lagler
(1962), Atlas of Fish Anatomy, Plate V, Michigan Fisheries No. 5, Department of Fisheries,
School of Natural Resources, The University of Michigan, Ann Arbor, MI.
-------
Urolithiasis (U) - This condition is known as nephrocalcinosis and involves
deposition of a white or "cream-colored" amorphous mineral material in the
tubules of the kidney. It can range in appearance from very small white spots
to severe involvement with very large "serpentine" deposits. These sites of
deposition are not to be confused with the Stannius bodies or corpora of
Stannius which are present in salmonid kidneys and have an endocrine function.
The Stannius bodies are generally not associated with the tubules and usually
occur at the "edges" in an area about midway along the kidney. They appear
more globular than do the urolithic deposits.
Other (OT) - This is used to class any aberrations which do not fit into the
above scheme. Record it as T and describe it in the remarks.
10.8.6 Liver
10.8.6.1 The appearance of the liver can very well be an artifact of the
sampling and the investigator should take that into consideration. Appearance
may, for example, vary with the length of time from collection to observation.
It also depends to a certain extent on the nature and extent of the loss of
blood during sampling. For this reason, categories "A" and "B" are both
considered as normal.
A - Normal. Good solid red color.
B - Lighter or less vivid red color than in A. Not so pale as to be classed
as general discoloration. Still considered to be normal.
C - "Fatty" liver. Light tan color, such as "coffee with cream".
D - Nodules in the liver, i.e., white mycobacterial cysts and incipient
nodules, such as those in hepatoma.
E - Focal discoloration. Color change in the whole liver.
OT - Aberration or deviation in liver which does not fit into above scheme.
Class as OT and describe in remarks.
10.8.7 Bile
10.8.7.1 The bile is observed indirectly through observation of the color of
the gall bladder. The ranking scheme considers "fullness" of the bladder and
degree of "green".
0 - Yellow or straw color; bladder empty or only partially full.
1 - Yellow or straw color; bladder full, distended.
2 - Light green to "grass" green.
3 - Dark green, dark blue-green.
10.8.8 Sex
254
-------
10.8.8.1 Observation of the gonads when possible should permit determination
of gender of the fish. It is also recommended that a remark be entered if the
fish are "ripe" or approaching spawning condition.
Male (M) - Observation of testes
Female (f) - Observation of ovaries
10.8.9 General Observations and Remarks
10.8.9.1 Anything which appears to be abnormal should be noted. It is
recommended that the mesenteric tissues in the visceral cavity be checked for
hemorrhage and inflammation and if these conditions are present, they should
be so noted in general remarks.
10.9 Calculation and Summary of Fish Health and Condition Assessment
10.9.1 Now that the fish have been sampled, examined, and as the observations
have been made and recorded on the worksheets, all the necessary calculations
should be made and summarized.
10.9.2 The format for "Summary of Fish Necropsy" is presented in Table 3.
That form will be used for the purpose of this discussion. The section
dealing with the heading information will be discussed in a later section, as
will the use of the computer. It is more than helpful to use a pocket
calculator which is provided with a function for standard deviation.
Ktl - The values of "K" (= coefficient of condition for the metric system)
have been used widely by fishery biologists to express the relative robustness
of fishes. Also, the values of "K" have been used additionally for age and
growth studies to indicate the suitability of an environment for a species by
a comparison of the value for a specific habitat with that of other aquatic
habitats. The value for Ktl is actually expressed here as Ktl x 105. This
was done to mitigate the problem of carrying a large number of decimal places
in the records. The equation used to obtain the value is as follows:
Ktl x 105 =
W x 105
L3
Where W = Weight in grams
L - Total length in millimeters
10.9.3 The condition factor used in the English system is Ctl. This value
tends to be used by some fish culturists. Ctl x 104 is obtained by
multiplying (Ktl x 105) bu 3.613.
10.9.3.1 The mean, standard deviation and coefficient of variation are to be
calculated for the length, weight, Ktl, hematocrit, leucocrit, and plasma
protein.
255
-------
Mean - The mean is determined by totaling all of the values for the
observations and dividing by the number of the observations.
Standard deviation - Indepth discussion of the standard deviation is beyond
the scope of this presentation. A pocket calculator equipped with a standard
deviation function permits very easy determination of that value. To
calculate the value without the aid of such a tool would require a prohibitive
amount of time.
Coefficient of variation - This value is defined as the ratio of standard
deviation to the mean. To obtain this value, divide the standard deviation by
he mean and multiply by 100 to convert the answer to percent. This value
expresses variation as percent of the mean. Units are not used. Record the
results on the necropsy summary sheet.
10.9.4 Values As Percents Of Total Sample
10.9.4.1 This portion expresses the percent of the total sample constituted
by each category. As an example, you can consider the eyes. The number of
fish with normal eyes divided by the total number of fish in the sample yields
the percent normal and should be recorded. The percent of fish with one blind
eye (Bl) is calculated in the same manner and so on. This is repeated for
each category of organ or tissue observation and results are recorded on the
necropsy summary sheet.
10.9.5 Summary of Normals
10.9.5.1 This section of the necropsy summary is included to facilitate
easier reading with respect to departure from normal. This also facilitates a
more accurate summary for those organs and tissues with more than one category
considered to be normal, i.e., liver and spleen. It must be further noted
that "0" is considered to be normal with respect to degree of hemorrhage in
the thymus and degree of inflammation in the hind gut. "N", when present, is
understood to be normal and the percent of the sample is indicated in the
value distributions. In these instances, merely carry that figure down to the
summary of normals. In the following instances the "normal" is not so readily
apparent:
Spleen - Black, red, and granular are all considered to be normal
manifestations of spleen condition. If the sample demonstrated 701 black, 15%
red, and 15% granular, you would combine these and list 100% normal in the
summary tables.
Liver - The A and B categories are both considered to be normal. Combine
these normals in the summary or normals.
Thymus - The categories included in the observation of the thymus represent
degree of petechial or "pin-point" hemorrhage. It is, therefore, understood
that "0" hemorrhage is normal. The percent of fish with "0" thymus is carried
down to the section dealing with summary of normals.
Hindgut - Degree of inflammation is being measured here so a reading of "0",
256
-------
TABLE 3. SUMMARY OF FISH NECROPSY
LOCATION:
SDacies:
Strain:
Mark/Lot:
Autopsy Date:
Hatching Data:
QUALITY CONTROL #
Sample Size:
Aqa:
Unit Case History #:
Fish Source:
Egg Souree;_
Water Temp.:_
Water Hardness:
lnvestigator(s}_
Reason for Autopsy:_
Remarks:
MEAN
Ltngth
Weight
Ktl*
cn**
Hematoorit
Leucocrit
Plasma Protein
STANDARD DEVIATION
COEFFICIENT OF
VARIATION
•Expressed as Ktl times 10 to the fifth power
**Converted from Ktl: expressed as Ctl times 10 to the fourth power
VALUES AS
EYES
N
B1
B2
E1
E2
H1
VS.
Ml
M2
OT
GILLS
N
F
C
M
P
OT
PSEUD
N
S
L
S&L
1
OT
THY
0
1
2
-
x«
MES
FAT
0
1
2
3
4
-
x =
PERCENTS OF TOTAL SAMPLE
SPL
B
R
G
NO
E
OT
HIND
GUT
Q
1
2
-
x =
KID
N
S
M
G
U
OT
UV
A
B
C
D
E
F
OT
BILE
0
1
2
3
-
x=
FIN
0
1
2
-
x =
OPER
0
1
2
.
x= ,
SUMMARY OF NORMALS
r
i
xxxxx
xxxxx
SUMMARY OF MEANS
xxxxx
xxxxx I
xxxxx I
1 XXXXX
1 xxxxx 1
xxxxx 1
.SEX
M:
Fat Index
Bile Index
Thymus Index
INDEX SUMMARY
Gut Index
Opercla Index
Fin Index
Normality Index
Severity Index
GENERAL REMARKS
FINS:
SKIN:
GONADS:
OTHER:
257
-------
indicating no inflammation, would be considered to be the normal. The percent
of the sample with "0" is carried down to the summary of normals.
Fins - Degree of active erosion is being measured here so a reading of "0",
indicating no active erosion would be considered to be normal. The percent of
the sample with "0" is carried down to the summary of normals.
Opercles - The relative degree of shortening of the opercles is being assessed
here so a reading of "0", indicating no "shortening", would be considered
normal. The percent of the sample with "0" is carried down to the summary of
normals. Mesenteric Fat and Bile - There are no normal categories for
mesenteric fat deposit and bile.
10.9.6 Summary of Means
10.9.6.1 This Subsection deals only with categories quantifying relative
degrees of some manifestation. Those categories involved in this section are
thymus, mesenteric fat depot, hind gut, and bile. This appears to be
confusing to people but the means are obtained in the usual manner. Total the
values in the appropriate columns and divide that total by the number of
observations. The "x" listed in the summary section dealing with values as
percents of total sample is the mean of the values and should be carried down
to the summary of the means. Numerous investigators using these systems have
referred to these means as indices, i.e., thymus index, fat index, etc.
10.9.7 Index Summary
10.9.7.1 The fat index and the bile index are the same as the means for those
observations as listed in the summary of means. The thymus, gut, fin, and
opercle indices are calculated by dividing the mean (listed in the "summary of
means") by the highest level possible and multiplying it by 100 to express it
as a percent. If, for example, the thymus mean would be .75, one would divide
this by 2 (the highest level possible) and multiply by 100 to yield 37.5
percent. This then becomes the thymus index. The severity index is
calculated by averaging the thymus, gut, fin, and opercle indexes. The
normality index is calculated by averaging the normals as listed in the
summary of normals. All of the indices are to be placed in the index summary
of the report for clarity.
10.9.8 Miscellaneous Observations
Sex - The relative proportion of gender should be entered if that information
is available. Here, as above, merely count the numbers of each category and
divide by the number of fish in the sample. If the investigator(s) is unable
to determine the gender, be sure to enter "U" for unknown.
General remarks - Any remarks made in the remarks column of the worksheet and
any general remarks, the investigator wishes to make should be made in this
section. There is a great deal of latitude here. One might, for example,
list under "Fins" that 10 fish or 50 percent of fish had badly eroded,
bleeding pectoral fins.
258
-------
10.9.9 Heading Information
10.9.9.1 The information entered into the heading of the worksheet and
summary is very important. It is that information which identified the
investigation and which ties it into the greater data base which will permit
future recall, manipulation, etc. It is very important that standard
terminology, abbreviations, ID systems, and cross-referencing be developed and
used to facilitate use in a data base. This is particularly true where
computers are to be used. It is likely that even more information will be
saved in relational data bases to enhance the value of the information. It
should be remembered that the worksheet and necropsy summary were developed to
be used both in hatcheries and free-ranging populations. This is in evidenced
more in the heading information than in any other portion of the
investigation. Many of the categories are self-explanatory, but some are
confusing enough to require a brief description. The following is a list of
categories with brief statements on some of the less obvious:
Location - Site or location of the study, such as Midway Hatchery or Green
River.
Quality Control Inspection No. - This is the number assigned to this
particular investigation.
Species - Species of fish being investigated. If abbreviations are to be
used, they should be standardized, i.e., RBT for rainbow trout.
Strain - Strain of fish under investigation, i.e., Sand Creek.
Necropsy Date - Date the necropsy was performed.
Sample Size - Number of fish in that particular sample.
Age - Age of fish using standard expression, such as months.
Mark/Lot - Identifying mark, such as dye mark or fin clip in free-ranging fish
or a production lot number in a hatchery.
Unit - Raceway number in a hatchery or specific station location, such as
Little Hole, Green River.
Water Temperature - The temperature of the water at the sampling site.
Fish Source - This generally refers to the original source of fish. The
investigation may be on fish in the Green River, but they may have been
stocked by a hatchery. The hatchery would be listed as the fish source in
this case. If they were natural reproduction, the Green River would be listed
as the fish source.
Egg Source - This refers to the original source of the eggs. In the example
above, the eggs may have been shipped to the hatchery by a brood station at
some other location. That brood station would be listed as the egg source.
259
-------
Water Hardness - This is expressed as parts per million (ppm).
Investigator(s) - Name of all investigators.
Hatching or Station Date - The date fish samples for collected.
Reason for Necropsy - Indicate reasons; such as research, routine, trouble
shooting, etc.
Remarks - Any information which might have an effect on interpretation of
results, i.e., fish were electro-shocked and hauled in tub for half an hour or
fish were taken in an overnight gill net set.
Tissue Collection No., Disease Survey No., Case History No., and Custody No. -
These are all cross-references to other investigations which should be carried
in the data base and which might have bearing on interpretation of results.
Purpose Code - Relates somewhat to "reason for Necropsy". It is included
because it makes it possible to do better sorts and queries later when working
with the assimilated database. It is very important that his be filled in. A
single letter coded is used as listed below:
A = Routine quality control inspection
B = Prestocking quality assessment
C = Trouble shooting
D = Research or special investigation
E = Administrative request for quality control
0 = Other, make entry in Remarks area
10.9.9.2 Other letters will be included later as we add letters more relevant
for fisheries biologists. This is why "0" is used for "other" rather than
"F". It is possible in this case to use more than one letter in combination
if it seems necessary. It may, for instance, seem appropriate to use AB
because the last routine quality control inspection may also be a prestocking
quality assessment and may be important in the use of the accumulated
database. All of this will be even more useful when viewed along with "Reason
for Necropsy" above,
10.9.9.3 The importance of the heading information cannot be overstated. It
is not uncommon to find that individuals have not been as diligent as they
might have been in achieving this portion of the investigation. It requires
only a few minutes more and makes a difference in the preparation of the
results. It is also necessary to the retrieval and manipulation of
information in data bases. This permits it to move from project significance
to program significance.
260
-------
10.9.9.4 Once completed, the necropsy summary presents a fish health and
condition profile of the population of fish sampled (see Tables 4 and 5;
Subsection 10.10.2).
10.9.10 Computer
10.9.10.1 This system lends itself very well to spreadsheet analysis and data
base management. A computer program has been developed for calculation,
summary, and reporting of the fish health and condition assessment necropsy.
AUSUM is a template for Lotus l-2-3R. It requires a copy of Lotus 1-2-3 ,
version 2.0 or later and an IBM compatible PC with at least a 512 K memory.
The report is formatted in such a way that the printer must be capable of 12
characters per inch and 8 lines per inch. It is a very user-friendly
template. The computer program is not necessary to use this methodology, but
it makes the task much easier, facilitates standard reporting, and provides
the basis for a data base. Instructions for using the AUSUM template are
given in Subsection 10.10, and a separate 30 page user's manual has been
prepared for the AUSUM 2.6 program and is available from Ronald W. Goede, Utah
Division of Wildlife Resources, Fisheries Experiment Station, 1465 West 200
North, Logan, UT 84321.
10.10 AUSUM 2.6--Computer Program for the Necropsy-Based Fish Health And
Condition Assessment System
10.10.1 INTRODUCTION
10.10.1.1 The computer program is written for Lotus l-2-3R, version 2.0. It
is a large worksheet so a computer with at least 512 K memory is needed. The
program calculates and summarizes the information and produces a printed
report. The printed report is formatted for 12 pitch and 8 lines per inch.
the printer should be capable of this or the report will not fit properly.
10.10.1.2 AUSUM is a computer program that has been specifically designed to
supplement the Necropsy-Based Condition Assessment System developed by Ron
Goede. The program, which is based on Lotus l-2-3R, provides a standard
report format and facilitates interpretation of the results. The following
features are provided:
* Menus for ease of use
* Defined format for data entry
* Capability to process 60 sample records
* Automatic calculation of the condition factor (Ktl) and all
summary information
* Summary information produced in report format
* Hardcopy of sample data produced for reference
* Ability to view Summary information prior to printing the report
Prepared by Ronald W. Goede and Sybil Houghton (1987), Utah Division of
Wildlife Resources, Fisheries Experiment Station, Logan, Utah 84321.
261
-------
TABLE 4. SAMPLE OF FISH NECROPSY COMPUTER SUMMARY REPORT I
SUMMARY OF AUTOPSY
LOCATION:
Midway
QUALITY CONTROL NO,: H22Y84
Species: CT
Strain: CTiL
Mark/Lot: 22-Y-8
Unit: 11 & 12
Fish Source: MM
Egg Source: BL
Hatching Date: 07/01/89
Remarks:
No unusual variables
Autopsy Date: 07/26/90 Sample Size
Age: 13 mos Tissue Collection Ma.
Disease Survey No.
Mater Temp.: 56 F Case History No.
Water Hardness: 550 pprn Custody No.
Investigator: Eric Purpose Code
Reason for Autopsy: Regular autopsy
HA
NA
MA
NA
A
Length
Weight
Ktr
Ctl**
Hematocrit
Leucocrit
Plasma Protein
MEAN
199.000 am
70.400 gr
0.890
3.215
37.900
0.880
4.130
STANDARD
DEVIATION
22.34 mn
25,67 gr
0.09
3.03
0.41
1.05
COEFFICIENT
OF VARIATION
11X
36X
US
8X
47X
25%
'Expressed as Ktl times 10 to the fifth power
"Converted from Ktl; expressed as Ctl times 10 to the fourth power
VALUES AS PERCENT OF TOTAL SAMPLE
PSEUDO-
MESEN.
HIND
EYES
N 100%
81 0%
82 0%
El 0%
£2 OX
HI OX
HZ OX
Ml 0%
MZ 0%
OT 0%
SILLS
N 100%
F OX
C 0%
H OX
P OX
OT 0%
8RANCHS
H 45X
S 55%
L 0%
S&L 0%
I OX
OT OX
THYHUS
0 90S
1 10%
Z 0%
x 0.1
FAT
0 OX
1 20X
2 20X
3 45X
4 15%
x 2.6
SPLEEN
8 20X
R 75X
S 5%
NO OX
E OX
OT OX
GUT
0 100X
1 OX
2 OX
x 0.0
KIDNEY
N 100%
S OX
H OS
G OX
u ox
OT 0%
LIVES
A 80%
B 20%
C OS
D 0%
E OX
F OS
OT OS
BILE
0 85%
1 15%
2 OS
3 OS
x 0.2
FIN
0 90%
1 10X
2 OX
x 0.1
OPERCLE
0 85X^
1 15X
2 0%
x 0.1
100%
100X
45%
Sumnary of Normals
SOX
100X
100% 1QOX 100X xxxxxxx
SSX
xxxxxxx
xxxxxxx
xxxxxxxx
Sumnary of Means
0.1
2,6
xxxxxxx
0.0
0.2
0.1
0.1
SEX: M: 65% F: 35X U: OX
GENERAL REMARKS
FINS Some upper caudals nipped
SKIN Clear
GQNADS Developing
OTHER 111. 12. 14. 15 twisted intestine
262
-------
TABLE 4, SAMPLE OF FISH NECROPSY COMPUTER SUMMARY REPORT I (CONTINUED)
Qual Uy Control No. M22Y84
SN LSH yGT Kt1 EYE GILL PS8R THY FAT SPL SUT KID LIV BILE SEX HEM LEU PLPR FIN OPCL
1
2
3
4
5
6
7
a
9
10
11
12
13
14
15
16
17
18
19
20
El
22
23
24
25
26
27
28
29
30
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
SO
242
173
211
187
183
193
203
222
180
198
178
189
210
203
143
185
230
215
223
212
119 0.84
41 0.79
72 0.77
60 0.32
52 0.85
65 0.90
70 0.84
88 0.80
53 0,91
72 0.93
35 0.62
50 0.74
93 1.00
64 0.77
22 Q. 75
57 0.90
122 1.00
97 0.98
96 0.87
80 0.84
N
H
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
M
N
N
N
N
H
N
N
N
N
M
N
N
N
N
N
N
. N
N
N
N
S
N
S
N
S
S
N
N
N
N
S
S
N
S
S
S
N
S
N
S
0
1
0
0
1
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
4
2
3
3
2
3
3
3
2
3
1
1
4
1
1
3
3
3
4
2
B
B
R
R
R
R
R
B
R
R
R
R
R
R
R
B
R
6
R
R
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
H
N
N
N
N
A
A
A
A
A
A
A
A
A
A
A
i
A
8
A
• 9
B
A
A
A
1
1
0
0
0
0
Q
0
0
a
0
0
i
0
0
0
0
0
0
a
H
M
F
F
N
N
F
F
F
N
H
M
M
H
F
F
N
H
H
N
36
35
38
44
37
41
35
44
40
39
39
33
37
37
36
31
35
36
40
39
1
1
1.5
0.5
0.5
1,5
0.5
1.5
0.5
0.5
0.5
0
1
1
1
0.5
1
1.5
1
1
3.8
4.8
3.8
6.1
3.3
4.6
4.0
5.1
4.0
4.3
2.2
3.2
4.8
3.1
2.0
3.7
4.2
6.0
5.1
4.5
0
0
1
0
0
a
0
0
0
0
0
1
0
Q
0
0
0
0
0
0
0
0
0
I
g
0
0
1
0
0
Q
0
0
0
0
1
0
Q
0
0
263
-------
TABLE 5. SAMPLE OF FISH NECROPSY COMPUTER SUMMARY REPORT I!
SUMMARY OF AUTOPSY
LOCATION:
fireen River
QUALITY CONTROL NO.: 88-23B
Autopsy Date: 7-6-
Age: 14 mos
Species: Cutthroat.
Strain: Bear Lake
Mark/Lot: 15Z6
Unit: Little Hole Water Temp,: 50
Fish Source: yhiterocks Water Hardness:
Egg Source: Egan Investigator: Barton,
Hatching Date: 4-23-87 Reason for Autopsy:
Remarks: Plasma samples: A403 to 414
Sample Size
Tissue Collection Mo.
Disease Survey Mo.
F Case History No.
260 ppm Custody Mo.
Purpose Code
Sreen River Project
HA
NA
NA
NA
0
60
Length
Weight
Ktl*
Ctl**
Hematocrit
Leucocrit
Plasma Protein
MEAN
222.330 nm
117.820 gr
1.070
3.866
40.710
1.690
6.660
STANDARD
DEVIATION
20.69 rim
39.81 gr
0.94
4.S9
0.51
0.72
COEFFICIENT
OF VARIATION
9%
34%
88%
12%
30%
11%
'Expressed as Ktl times 10 to the fifth power
**Cortverted from Ktl; expressed as Ctl times 10 to the fourth power
VALUES AS PERCENT OF TOTAL SAMPLE
EYES
N 100%
Bl OX
12 0%
El Q%
E2 0%
HI 0%
H2 0%
HI 0%
M2 0%
OT 0%
GILLS
N 100%
F 0%
C 0%
M 0%
P 0%
OT 0%
PSEUOQ-
BRANCHS
N 100%
S 0%
L 0%
S&L 0%
I 0%
OT 0%
THYMUS
0 43%
1 52%
2 5%
x 0.6
MESEN.
FAT
. 0 20%
1 40%
2 • 7%
3 25%
4 8%
x 1.6
SPLEEN
B 27%
R 73%
6 0%
NO 0%
E 0%
OT 0%
HINO
GUT
0 83%
1 17%
2 0%
x 0.2
KIDNEY
N 100%
S 0%
M 0%
6 0%
U 0%
OT 0%
LIVER
A 12%
B 88%
C 0%
D 0%
E 0%
F 0%
OT 0%
BILE
0 63%
1 30%
2 7%
3 0%
x 0.4
FIN
0 47%
1 35%
2 18%
x 0.7
QPERCLE
Q 77%
1 13%
2 1«
x 0.3
100%
100%
100%
Suimiary of Normals
43X xxxxxxx
100%
83%
100% 100%]xxxxxxx
47%
77%
Summary of Means
xxxxxxx
SEX:
xxxxxxx
M: 62%
xxxxxxxx
F: 38%
0.6
U: 0%
1.6
xxxxxxx
0.2
xxxxxxx
xxxxxx
0.4
0.7
0.3
GENERAL REMARKS
FINS Left pelvic fin clipped; avg. fin index = 0.7
SKIN Red dye marked
60NADS NA
OTHER 3 fish w/mild inflamation of hind gut
264
-------
TABLE 5. SAMPLE OF FISH NECROPSY COMPUTER SUMMERY REPORT II (CONTINUED)
Qual, Control No, 88-238
SN LGH VGT Ktl EYE GILL PSBR THY FAT SPL GUT KID LIV BILE SEX HEM LEU PLPR FIN OPCL
1
2
3
4
5
6
7
8
i
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
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
209
220
195
207
210
214
221
210
219
215
195
195
226
230
222
223
205
208
230
203
218
235
233
238
232
270
255
225
226
251
232
220
217
227
209
230
217
207
205
220
236
215
232
247
232
227
236
218
234
241
241
234
250
210
220
215
250
223
115
240
74 0.81
90 0.85
68 0.92
81 0.91
79 0,85
86 0.88
89 0.82
85 0.92
85 0.81
82 0.83
60 0.81
63 0.85
111 0.96
99 0.81
98 0,90
102 0.92
70 0.81
69 0.77
116 0.95
75 0.90
89 0.86
114 0,83
116 0,92
121 0.90
108 0.86
186 0.94
136 0.82
99 0,87
105 0.91
151 0.95
112 0.90
93 0.87
82 0.80
101 0.86
81 0.39
115 0.95
91 0.89
78 0,88
75 0,87
90 0.85
187 1.42
128 1.29
153 1.23
200 1.33
169 1.35
153 1.31
200 1.52
169 1.63
153 1.19
172 1.23
133 0.95
164 1.28
210 1.34
175 1.89
162 1.52
162 1.63
107 0.68
141 1.27
123 8.09
171 1.24
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
«
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
1
0
0
1
1
0
1
1
1
0
0
0
1
0
1
1
0
1
1
0
0
0
0
2
0
1
0
1
1
1
0
1
1
1
0
0
1
1
0
1
0
1
1
2
0
1
0
1
1
1
0
1
1
2
0
1
1
0
0
0
0
0
1
1
0
1
1
1
1
1
0
0
1
1
1
1
1
0
1
3
0
1
1
1
0
2
0
1
1
2
2
1
A
1
1
I
0
1
0
0
4
3
3
3
3
3
3
3
3
4
4
2
3
3
3
3
3
3
4
4
R
R
B
B
R
R
R
R
R
R
R
R
R
R
R
R
8
B
R
R
R
B
R
B
B
R
R
R
R
R
B
R
R
R
R
R
B
R
R
R
R
R
R
8
B
B
R
R
R
R
B
R
R
R
B
R
B
R
R
B
0
0
0
1
0
0
0
0
0
0
0
0
0
0
0
1
0
1
0
I
0
0
1
0
0
0
0
0
0
0
0
1
0
0
1
0
0
0
0
1
1
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1
N
N
N
N
N
N
N
N
n
N
N
N
H
N
N
N
H
N
N
N
N
N
N
H
N
N
N
N
N
H
N
N
N
H
N
H
H
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
H
N
H
8 .
B
8
8
B
B
B
8
8
B
8
8
3
B
8
8
8
B
B
A
8
3
B
8
8
8
6
B
B
B
B
B
8
B
8
8
8
B
8
B
B
A
B
8
A
8
B
B
B
A
B
B
%
8
8
A
B
B
A
A
0
0
1
1
0
0
0
1
1
0
0
0
0
1
1
1
0
0
1
1
0
0
0
1
0
0
0
0
0
0
0
1
0
0
1
0
0
0
0
1
0
0
2
2
0
1
1
1
2
0
0
0
2
0
0
0
0
0
1
1
F
H
F
M
H
M
H
M
F
F
M
H
H
M
F
F
M
F
F
M
M
F
H
M
F
M
F
F
F
M
H
F
M
M
F
H
F
F
M
F
M
M
F
H
F
N
F
M
H
M
N
H
M
M
F
M
M
H
F
M
38
44
42
46
42
40
41
38
42
45
41.5
37
38
41
36
40
52
47
36
41
37
38
34.5
36
33
42
42.5
36.5
40
35.5
38
35
37
37
37.5
33
34
34
41
41
46
49
41
41
49
46
40,5
44
42.5
45
39
41
30
44
43.5
46
50
49
45.5
1
1.5
1.5
1
1.5
1.5
2
2
2
1.5
2
2
2,5
2
2
1
1,5
1
1.5
2
2.5
2.0
2.5
2.0
1.5
2.0
2.0
2.5
2.5
2.0
2.0
2.0
2.0
1.5
2.5
1.5
2.0
2.0
1.5
2.0
0.5
2.0
1.0
1.0
1.0
1.0
0.5
1.0
1.5
2.0
2
2
1.5
1
1
1.5
2
2.5
1.5
6.8
7,1
6.1
7.3
6.0
6.5
7.0
6.8
6.1
6.4
5.7
7.1
6.6
5.8
6.0
7.0
6.9
6.1
6.0
6.7
6.3
6.6
6.2
6.3
6.1
6.0
6.5
6.4
7.0
6.7
6.0
5.9
5.5
6.5
7.1
5.0
6.5
7,1
6.7
6.3
7.9
7.1
6.8
7.0
9.4
8.1
8.1
7.0
7.2
7.1
6.7
6.3
5
7.3
7
7.1
7.4
7.1
6.8
2
2
1
0
2
0
1
0
2
1
1
0
1
0
0
2
2
0
2
2
0
1
0
1
1
2
1
1
0
1
1
2
1
1
1
1
2
1
1
1
0
1
0
0
0
0
0
0
0
0
0
0
0
0
0
1
0
0
0
0
1
1
1
1
1
2
2
2
2
1
2
1
2
1
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
a
0
0
0
0
0
0
0
0
0
265
-------
Prior to use of AUSUM, the following Subsections should be read by all
users:
INTRODUCTION
COMPUTER REQUIREMENTS
BEGINNING STEPS
PRINTER SETUP
MENU PRINTER
MACRO PRIMER
AUSUM PROGRAM USE
OTHER PROGRAM SELECTIONS
10.10.1.3 Keyboard Primer (page 279) is provided for those who are not
familiar with computers. Lotus Primer (page 280} gives background information
for those who are unfamiliar with Lotus l-2-3R. Entry Requirement (page 284)
lists the data-entry requirements. Sample reports are provided (see pages
262-265, 286-287).
Keyboard Primer is provided for those who are not familiar with
computers. Lotus Primer gives background information for those who are
unfamiliar with Lotus l-2-3R. Entry Requirements lists the data-entry
requirements.
COMPUTER REQUIREMENTS
AUSUM has been designed for the following computer configuration:
2 floppy disks
IBM PC or compatible with at least 512 K memory
Lotus l-2-3R, version 2.0
Epson dot matrix printer (see Printer Setup, page 267) for instructions to
change the print setup to accommodate other printers.
BEGINNING STEPS
The AUSUM master disk is to be kept for backup purposes only. Before
using AUSUM, you need to copy the program onto your own formatted disk. You
will also need a formatted disk for a data disk. Use the following
instructions to format your disks and copy the program disk:
A. Format a new disk.
1. Place the DOS system disk in drive A.
2. Place the new, unformatted disk in drive B.
3. At the A> prompt, type:
FORMAT B: (Then press Return key)
266
-------
B. Copy the AUSUM master disk.
1. Place the AUSUM master disk in drive A.
2. Place a formatted disk in drive B.
3. At the A> prompt, type:
COPY *.* B: (Then press Return key)
4. Store the original AUSUM master disk in a safe, dry place.
This disk should never be used to run the program.
C. Follow Step A directions to format a new disk to be used for your
data disk.
PRINTER SETUP
AUSUM has been designed to use an Epson dot matrix printer. The reports
are designed to be printed using elite type (12 pitch), 8 lines per inch;
thus, the program uses the following command (setup string):
\027\077\0270
Should your printer need a different setup string for elite type, 8 Ipi,
you may use the Print Set option from the Submenu of the AUSUM program. You
will be asked to enter the elite, 8 Ipi, setup string for your printer.
Simply enter the correct setup for your printer, and the program will
automatically setup the printer command for you.
MENU PRIMER
There are two menus for AUSUM:
Main Menu and Submenu
To activate the Main Menu, press Alt-M. To go the Submenu, select the
Submenu option from the Main Menu.
Selections may be made from the menus by either of the following
methods:
(1) Press the beginning letter of the desired selection, such as H for
Heading
(2) Move the Control Panel cursor to highlight the desired
selection,then press ENTER.
The following is a brief description of the menu options:
Main Menu
Heading - Enter heading and general remarks
Data - Enter sample data
267
-------
Calculate - Calculate Ktl and summary data
Report - Print a report and hardcopy of the data
Xtract - Extract data and heading for later use
Prepare - Prepare worksheet for new data entry
Load - Load previously saved data file
Submenu - Unlock, Printset, Extract-Edit, End Lotus, List files,
Summary, and Main Menu
Submenu
Unlock - Unlock titles
PrintSet Set elite command for your printer
Main Menu - Return to Main Menu
End - End work with Lotus/return to MS-DOS
Summary - To view summary information
List List files on data disk
Xtract-Edit - Extract edited data using previous or new file name
CAUTION: Prior to using the menus, you must be certain to deactivate any
commands that are currently in use; in other words, the status indicator CMD
must not be showing at the bottom of the screen. (To deactivate the CMD,
press Ctrl-Break and the ESC.)
MACRO PRIMER
In Lotus l-2-3R it is possible to program a set of commands. These
programs are called macros. There are four macros which you will be using
while entering the processing the Necropsy (Autopsy) System data. Each of
these macros is invoked by pressing the Alt key simultaneously with the letter
that names the macro. For instance, to bring the D macro, press Alt-D. The
following is a list of the AUSUM macros, directions for their use, hints about
when you will utilize them, and directions to end them:
M - This macro brings the AUSUM Main Menu Control Panel area (top
portion) of the screen. (See the Menu Primer, page 267, for an
explanation of the menu options.) Use the menu whenever you need
to select the next processing step. Press ESC to deactivate the
Main Menu. Press ESC twice or press Ctrl-Break and press ESC to
deactivate the Submenu.
D - This macro automatically shifts the cursor down to the next cell
whenever ENTER is pressed. You will want to use this when
entering the Heading Data and any columns in the Sample Data where
the entries vary down the column, such as lengths or hematocrits.
To end this macro, press Ctrl-Break (you will hear a beep) and the
ESC.
C - This macro permits you to copy a specific cell entry to a
specified range. You will want to use this when an entire column
is all the same entry, such as all N for Eyes. To use this macro,
do the following:
268
-------
(1) Place the cursor on the cell which contains the data
to be copied,
(2) Press Alt-C.
(3) Notice a message on the Control Panel will say:
Enter range to copy FROH:
Following the colon will be the current cell location,
repeated twice, such as A23..A23.
(4) Press ENTER.
(5) The message will now say:
Enter the range to copy TO:
After the colon, the current cell location will again
be repeated twice. (CAUTION: Be sure NUM LOCK is off
before using the arrow keys to highlight the copy
region.) Press the down arrow key to go down the
column as far as you want to copy the data. Notice
that the copy range is now highlighted. Also notice
that the second cell location on the Control Panel has
changed as you have moved the cursor. After the
desired range is highlighted, press ENTER. HINT; If
you desire to have two or more columns next to each
other with the same entry, such as two columns of N,
then highlight both columns by pressing the
appropriate arrow keys.
(6) The macro ends itself with no further entry needed
from you.
This macro will erase a specific range—or even just one cell.
This macro must be used with extreme caution because you want to
erase only incorrect data. To use this macro, do the following:
(1) Place the cursor on the cell to be erased or on the
top left corner cell of the range to be erased.
(2) Press Alt-E.
(3) Notice a message on the Control Panel will say:
Enter range to erase:
Immediately following the colon will be the current
cell location.
(a) If one cell is to be erased, press Enter.
269
-------
(b) If a range is to be erased, use the appropriate
arrow keys to highlight the range. Be sure you
want to erase a]], the highlighted area! Press
ENTER.
(4) The macro will end itself with no further entry needed
from you.
HINT; What to do if you begin a macro and something is wrong? You may
have entered a wrong character or the mode indicator says ERROR. To end a
macro at any time, press Ctrl-Break (you may hear a beep) and then pres ESC.
Of the ERROR message shows, you will probably only need to press the ESC key,
HINT; Lotus l-2-3R will not permit you to use more than one macro at any
one time. You will need to deactivate the menu or any other macro before
activating a new macro.
AUSUN PROGRAM USE
Program Startup
(1) Start the computer and load with MS-DOS 2.0 or later
version.
(2) Insert the Lotus l-2-3R system disk in drive A.
(3) At the A> prompt, type 123 and then press ENTER.
(4) As soon as the Lotus l-2-3R program is loaded (The worksheet
format will show on the screen), remove the Lotus 1-2-3
system disk and insert your copy of AUSUM in drive A.
(5) Insert the formatted data disk in drive B.
(6) To being the program, type: /FR (The file name, AUSUM,
will be highlighted on the third line of the Control Panel).
(7) Press ENTER. The screen will then appear as Figure 1.
(8) Press ENTER as directed, and the screen will then appear as
in Figure 2.
270
-------
Figure 1. Introduction to AUSUM
AUSUM
Version 2.6
Developed December 1986
by
Ron Goede and Sybil Houghton
If you have questions, contact:
Ron Goede
Utah Division of Wildlife Resources
Fisheries Experiment Station
1465 West 200 North
Logan, UT 84321
(801) 752-1066
Copyright Ronald W. Goede, Sybil Houghton - 1987
Press ENTER to continue . . .
Figure 2. Continuation of Introduction
AUSUM is used to summarize data from the most
recent version of the necropsy (autopsy) system which
includes observations of bile but not mesentery.
NOTE; AUSUM is not be used for data which
include observations of mesentery.
Press ENTER to continue . . .
(9) Press ENTER as directed, and the screen will then appear as
in Figure 3.
Figure 3. Data Disk Drive Entry Screen
On the line at the top of the screen, enter the
drive in which data disk is to be placed . . .
Then press ENTER to continue . . .
271
-------
(10) Enter the letter for the drive in which the data disk is to
be placed, (for a configuration with two floppy disk
drives, you will enter B for the drive letter.)
(11) Then press ENTER to continue. The screen will then appear
as in Figure 4. You are now ready to begin entering the
Heading information. See Program Order (page 273) for steps
to follow when using the AUSUM program. The cursor is
already located for the first entry.
Figure 4. Beginning Screen
D67: [W10]
A B C D
64 Enter the heading data
65 using the specified
66
67 Location:
68 Species:
69 Strain:
70 Mark/Lot:
71 Unit:
72 Fish Source:
73 Egg Source:
74 Date of Hatching"
75 Remarks
76 Necropsy Date:
77 Age:
78 Water Temp. :
79 Temp. Scale (C or F) :
80 Water Hardness:
81 Investigator:
82 Reason for Necropsy
83 Qual. Control No.:
84 Sample size
85 Tissue Collection No.:
86 Disease Survey No.:
87 Case History No. :
88 Custody No.:
89 Fins:
90 Sins:
91 Gonads:
92 Other:
93 Purpose Code:
READY
E
in column D
field lengths { }:
{30}
{13}
{13}
{'13}
{17}
{8}
{8}
{'MM-DD-YY)
{68}
{'MM-DD-YY}
{10}
{2}
{1}
{4}
{15}
{30}
{'7}
{2}
{'7}
{'7}
{'7}
{'7}
{65}
{65}
{65}
{65}
{2}
272
-------
Program Notes
Before you begin to use the program, read the following notes:
(1) When you begin the program » the cursor is already in
position for you to enter the heading data.
(2) You are instructed to enter the data column D according to
the specific directions given. There are three types of
directions:
(a) {'MM-DD-YY) Enter dates, such as '12-06-86. You
must use the apostrophe (') in front
of the date. (For explanation, see
Label/Value section of the Lotus
Primer, page 280.)
(b) {13} The number (13) indicates the
maximum number of characters
al1 owed.
(c) {'7} A number used as a label. You must
use the ', The number (7) shown
indicates the maximum number of
characters allowed in addition to
the apostrophe.
Example: '86-02-1
(3) When the Main Menu us activated, the selections will be
displayed on the Control Panel (top portion of the screen).
Program Order
The usual order of menu selections when entering a set of data for the
first time is:
(1) Heading
(2) Data
(3) Calculate
(4) Report
(5) Xtract
(6) Prepare
Heading
To enter the Heading information, use the following directions:
(1) Invoke the Down macro by pressing Alt-D.
(2) Enter the information in the appropriate cells.
(3) If there is no information for a particular cell.
273
-------
To correct entries, do one of the following:
(1) Use the down or up arrow keys to move to the appropriate
cell. They type the correct entry.
(2) If you desire to EDIT the entry, do the following:
(a) Move to the appropriate cell.
(b) Deactivate the Down macro by pressing Ctrl-Break and
then ESC.
(c) Press the F2 key to EDIT.
(d) Edit the entry line.
(e) Press ENTER.
After all the Heading information has been entered, do the following:
(1) Deactivate the Down macro by pressing Ctrl-Break and then
ESC.
(2) Activate the Main Menu by pressing Alt-M.
Data
After you select Data from the Main Menu, the cursor is located in the
first cell of the length column. In this area of the worksheet, you may want
to enter data in either of the two following ways:
(1) Use the Down macro (see page 268) and enter data in the
individual cells as you go down the column.
(2) Use the Copy macro (see page 268) if the column entries are
all the same.
NOTE; The first cell of the Ktl column says ERR. This is not. a mistake or
error! The cell contains the formula to calculate the Ktl. During the
calculation process, the formula will be copied down the column and the Ktl
will be calculated for each item in the sample. Thus, no entry is required
for the Ktl column. (The Ktl column is not protected; thus, be careful that
you do not enter the data in that column.)
To help you with the data entry, the column titles and sample numbers
have ben "locked" in place. Thus, as you work you way down and across the
worksheet, you will always know the title of the column and number of row for
your current cell location.
Enter all the sample data before doing any calculations. After all the
data is entered, deactivate the Down macro, if necessary.
274
-------
REMEMBER;
Calculate
(1)
(2)
(3)
Report
(1)
(2)
(3)
(4)
The program is designed for a maximum sample size of 60.
Activate the Main Menu (Alt-M).
Select Calculate—the calculations will take a minute or so
to complete; thus, the screen will say: Please wait . , ,
At the end of the calculation process, the Main Menu will
again be displayed and you will be asked to make your next
selection.
Be sure the printer is on!
Select Report from the Main Menu.
On the Control Panel will be a question:
Has the printer been turned on? (0 or 1)
After checking to see that the printer is turned on, press 1
and the program will continue. If you decide not to print
the report, press 0 (zero) and you will be returned to the
Main Menu for your next selection.
A second question will then be shown:
Has all the data been entered? (0 or 1)
If you press 0 (zero), the Main Menu will be displayed so
you may make the appropriate data entry selection. If you
press 1, the program will continue to execute the print
commands.
(5) The screen will say:
Please wait
The standard formatted report and a hardcopy of the data
will then be printed.
(6) At the end of the printing process, the Main Menu will again
be displayed and you will be asked to make your next
selection.
NOTE: If you want to save the data on the data disk, you
must continue with the next step (Xtract); if not, the data
will be permanently lost.
275
-------
Xtract
By selecting this option, you will be saving (Xtracting) only the
heading information and the sample data rather than the entire worksheet.
(The program has been designed in this manner to conserve space on your data
disk). This selection is only for the first time you save (Xtract) the
specific set of data. (See Xtract-Edit for edited data, page 278).
To save your data entry on your data disk, do the following:
(1) Select Xtract from the Main Menu.
(2) The screen will show:
ENTER THE NAME OF THE FILE TO BE EXTRACTED . . .
(3) Enter the file name (limited to 8 characters) you wish to use for
this set of data. As you enter the file name the *.wkl will
disappear from the Control Panel and the file name will appear.
HINT: For easier file name recognition, we suggest you use the
specific Quality Control No. (i.e., 87-01) as part of the file
name, such as 87AU01. (You cannot use hyphens in a file name.)
(4) At the end of the extraction process, the Main menu will again be
displayed and you will be asked to make your next selection.
Prepare
This process will clear the worksheet and prepare it for a new set of
data. [CAUTION; Be sure you have saved (using the Xtract option) your data
before selecting the prepare option!]
(1) Select Prepare from the Main Menu.
(2) No questions to answer--just wait until it is complete.
(3) At the end of the preparation process, the Main Menu will again be
displayed and you will be asked to make your next selection. You
are now ready to enter a new set of data or load in a previously
saved set of data.
HINT; If you are running short on time and do not want to wait
for the printer to print the report, or if a printer is
unavailable, you may want to skip the Report option and just
Xtract (save) the data for now. Then at a later time you may load
the data and select the Report option.
276
-------
OTHER PROGRAM SELECTIONS
List
An additional feature which AUSUM offers is the ability to list the
files on your data disk. This List option is helpful for several reasons.
First, you may need to know whether the data disk is full before trying to
save a new set of data. (A diskette will hold approximately 25 extracted
necropsy (autopsy) data files.) Second, it will help you remember the name of
the data file that you want to load. To use the List option, do the
following:
(1) Select List from the Main Menu.
(2) A list of the files on the data disk will be displayed on the
screen.
(3) To end viewing of the file list, press ENTER.
Load
To process and/or edit data that you previously saved, you will need to
load that data into the worksheet, fCAUTION; Be sure that the worksheet is
prepared for new data prior to using the Load option.] REMEMBER: You may
select the List option to review the names of your data files prior to
selecting the Load option.
Place the specific data disk in the drive you selected for the data disk
at the beginning of the AUSUM program, and then do the following:
(1) Select Load from Main Menu.
(2) On the screen will be:
ENTER THE NAME OF THE FILE TO LOADED . . .
(3) Type in the appropriate file name.
(4) Press ENTER.
(5) After the data is loaded, the Main Menu will again be displayed
and you will be asked to make your next selection.
You may now do any necessary editing using the methods to correct
entries described in Heading (page 273) and Data (page 274). The program may
then be continued as if it were the original data entry. [CAUTION; Be sure
to select Calculate after editing and before a report, is printed. Calculation
must be performed each time you re-enter a file and make any changes.]
277
-------
Unlock
While you are entering the Sample data, the column and row titles are
locked into place. To deactivate the locking process, simply select the
Unlock option of the Submenu and press ENTER.
End
When you have completed your data entry for AUSUM and are finished with
your use of Lotus 1-2-3, select the End option from the Submenu. This will
return you to A> prompt of MS-DOS at the system level. [CAUTION: Be sure you
have saved all your data before you use the End option.]
PrintSet
See Printer Setup for an explanation.
Xtract-Edlt
When saving (Xtracting) data that has been previously saved, you must
use the Extract-EdIt option--,not the Xtract option. To help your memory, you
will be reminded of the name of the file which you have been editing. To use
this option, do the following:
(1) Select Xtract-Edit from the Submenu.
(2) On the screen will be:
THE NAME OF THE FILE YOU HAVE LOADED IS:
PLEASE ENTER THAN FILE NAME ... OR YOU MAY CHANGE
TO A NEW FILE NAME . . .
(3) Type in the appropriate file name.
(4) Press ENTER.
(5) At the end of the process, the Submenu will be displayed and you
will be asked to make your next selection.
Summary
This option allows you to view the Summary information. You may want to
use this option to check the information prior to printing the report. To use
this option, do the following:
(1) Select Summary from the Submenu
(2) On the screen will be some of the Summary information. Use the
arrow keys to view all of the information.
278
-------
KEYBOARD PRIHER
You will notice that the keyboard is very similar to that of a
typewriter. However, there are some additional keys. A brief description of
these additional keys follows:
Functional keys
On the left side (or across the top) of the keyboard are at least 10
keys which are labeled as Fl, F2, etc. These keys are pre-programmed by each
computer program to have specific capabilities. The only Function key you
need to use for this program is the F2 key, which is the Edit key.
Ctrl (Control) Key
This key is used in conjunction with other keys to enact specific
directions. An instruction such as Ctrl-Break means to press the Control and
Break keys simultaneously.
Scroll Lock/Break Key
This key is used when the instructions call for the Break key. It is
used in conjunction with the Control key to abort certain operations in Lotus,
The key has many other uses, but that is the only one you will be using for
this program. CAUTION; If you do not hold the Ctrl and Break keys down
simultaneously, the indicator SCROLL may appear at the bottom of the screen.
If this happens, press only the Scroll Lock/Break key to erase the SCROLL
indictor and then press Ctrl-Break simultaneously.
Alt Key
This key is used in conjunction with any letter key to invoke Lotus
macros (programs). For example, Alt-D means to simultaneously press the Alt
Key and the letter 0. By doing so you would invoke a macro identified by the
letter D. Refer to the Macro Primer (page 268) for a further explanation.
Number Pad
These keys permit you to efficiently enter numeric data. To invoke the
number pad, press the MUM LOCK key. [CAUTION; If the number keys have arrows
on them, they can be used only as numbers when the MUM LOCK key has been
pressed.
The NUM LOCK key is a toggle key; thus, to return to arrow or direction
use, press the NUM LOCK key again.
Arrow Keys
Your keyboard may have separate keys with arrows on them, or the arrows
may be on the number pad keys. (Be sure to read the caution included in the
Number Pad description above.) Use the arrow keys to move the cursor up,
down, right or left.
279
-------
HomeKey
This key is located with the arrow keys. While editing a cell entry,
you may use this key to go the beginning of the line being edited. [CAUTION;
Any other time the Home key is used, the cursor will be taken out of the
current position to the beginning of the screen. In that case you must return
to the menu (press Alt-M), then make you original selection and return to your
original position using the arrow keys,]
Del (Delete) Key
While editing a cell entry, you may use this key to delete the character
at the same location as the edit-line cursor,
Backspace Key
You may use this key while entering data or when editing. Pressing this
key will delete the character just to the left of the cursor location.
End Key
This key is located with the arrow keys. While editing a cell entry,
you may use this key to go to the right end of the line being edited.
ESC (Escape) Key
Use this key when you want to end an operation prior to its normal
completion. At times you will need to first press Ctrl-Break and then the ESC
key to end an operation.
LOTUS PRIMER
Introduction
Lotus l-2-3R is a spreadsheet-type of computer program. Such a program
is based on "cell entries." Picture the worksheet (working area of the
program) as a grid with columns named by letters and rows named by numbers.
Thus, each "cell" has a specific location such as Al or X36. (Perhaps you
have played the game "Battleship" that is based on this same type of grid
identification.) As you enter data in this worksheet, you will be filling a
cell with each "piece" of data.
Screen Format
An understanding of Lotus's screen format will be helpful. The Control
Panel comprises the top three lines of the screen. When you begin the
program, the Control Panel will appear as in Figure 4. The following example
is an explanation of the information on the first line:
Information Explanation
D67 Location of cursor
280
-------
{W10} Width of column
READY Mode of indicator
When using the menus, the selections will be displayed on the second
line. The third line will give the explanation for the highlighted menu
selection. Use the arrow keys to move the cursor across the second line, and
you will see that the third line changes to give the explanation of each menu
selection as it is highlighted.
Sometimes the second and third line of the Control Panel will be blank,
or there may be a question on the second line of the Control Panel that you
will need to answer. At other times you will need to enter the name of a
file. Further directives are given in the Menu Primer (page 267).
The lower left-hand corner of the screen, as shown in Figure 4, gives
the date and time. The remaining portion of the bottom line is used to tell
which "status indicators" are currently in use. This example shows CALC as
the current status indicator. While you are running the program, other status
indicators may appear, such as NUM, CMD, and CAPS.
The remaining portion of the screen is the actual worksheet area with
its column letters and row numbers for reference. All data entry will be made
by you in the worksheet area. This is more fully explained within the program
directions.
Label/Value
Typical of all computer programs, Lotus 1-2-3 has its own
idiosyncrasies. For data entry you must be aware of one particular Lotus
requirement. When you type the first character of an entry, Lotus immediately
determines whether the entry is going to be a VALUE or a LABEL. (The mode
indicator in the top right corner of the screen will change from READY to
VALUE or LABEL.) Sometimes this idiosyncrasy can present a problem. For
instance, you may want to enter a date as 12-06-85. Lotus assumes this to be
a value because the first character is a number. Thus, rather than displaying
your entry, Lotus would display -79, the result of 12 minus 6 minus 85!
Likewise, if you typed the date as 12/06/85, Lotus would display .02 which is
the result of 12 divided by 6 divided by 85!
Fortunately, there is a way to circumvent this "problem." You simply
need to begin this entry with an apostrophe, so you will enter '12-06-85. The
apostrophe tells Lotus that you want to treat these numbers as a LABEL rather
than as a VALUE. Note that as soon as you enter the apostrophe, the READY
mode indicator changes to LABEL.
Lotus considers all of the following as indicative of a VALUE entry:
0123456789 + -. $(
If you desire an entry that begins with one of these characters to be a
LABEL instead, you must begin the entry with an apostrophe.
281
-------
The slash (/) key is reserved for Lotus commands. You will not need to
use this key. In fact, it is recommended that you not used this key unless
you are familiar with the use of LOTUS. Should you accidentally press this
key, you may press the ESC key to negate its effect.
If the first character of an entry is other than those VALUE entries
shown above or a slash (/), Lotus assumes the entry us a LABEL. In this case,
you do not need to use the apostrophe--Lotus will automatically place it there
for you.
Now, what happens if you forget to use the apostrophe? One of two
things will happen:
(1) As in the date example above, Lotus will do the calculation
instead of accepting your entry as a LABEL. In such a case,
you may change the cell entry using either of the following
methods:
(a) Edit the cell entry.
(1) Press the F2 key.
(2) Press HOME to go the beginning of the entry
line.
(3) Press the apostrophe key.
(4) Press ENTER.
(b) Re-enter the entire cell entry using an apostrophe as
the first character.
(2) If you have combined number characters with label
characters, such as 80-6C, Lotus will beep and automatically
change to the EDIT mode. You may then simply press the HOME
key to go to the beginning of the entry line, press ', and
then ENTER.
Data Entry Methods
During the program you will be using two different methods for data
entry:
(1) To enter data in a single cell, do the following:
(a) Place cursor on cell where data in to be entered.
(b) Type the entry using an apostrophe where appropriate.
(c) Press ENTER.
This is the most efficient methods to use when
282
-------
entering the Heading information, length and wright
columns data, and all other columns where data varies
for each sample.
(2) If the entire column is all the same entry, such as all the
Eye entries are N or all the Thytnus entries are 0 (zero),
then it is more efficient to enter the desired character in
the first cell of the column and then copy this entry down
the column. To do this, use the Copy macro as explained in
the Macro Primer (page 268).
HINT; If only one or two of the column entries are different, you
may still prefer to use the Copy macro. After copying, go to the
one or two cells which should be different and enter the
particular data using the single cell entry method.
What do you do if you enter incorrect data? You may do either of the
following:
(1) For a single cell correction, move the cursor to the
appropriate cell and do one of the following:
(a) Type the entire entry again,
(b) Use the EDIT mode (F2 key) to correct the entry.
(2) For a block or range of cells (see definition of range
below), you may find it easier to erase the entire range and
then re-enter the data. Use the Erase Hacro to do the
erasure (see page 269).
Terminology
To help you understand and use AUSUM, the following Lotus 1-2-3 terms
are defined:
Cursor - There ar two types of cursors in Lotus 1-2-3:
(1) In the worksheet area, the cursor is a
highlighted area that designates the current
cell location. You will move this cursor with
the arrow keys when entering data.
(2) In the Control Panel, a blinking line underlines
the current location of the cursor.
Macro - A set of special commands that can be executed with
one key stroke combination: Pressing and holding down
the Alt key while at the same time pressing the key
representing the macro's name.
Mode - Displayed in the top right corner of the screen.
283
-------
Examples are READY, VALUE, LABEL, EDIT, AND MENU.
Hopefully, you will not have the ERROR mode! (If you
do, press ESC.)
Range - Specific area of the worksheet--one or more cells. It
must be a rectangle or square.
Worksheet - The screen area, except of the Control Panel (top
three lines) and the status indicator line (bottom
line). This is the work area for a Lotus program.
ENTRY REQUIREMENTS
Below is a list of the correct entries to be used for the AUSUM program:
ENTRY EXPLANATION ENTRY EXPLANATION
N
Bl
B2
El
E2
HI
H2
Ml
M2
OT
N
F
C
M
P
OT
N
S
L
S&L
I
OT
Normal
One blind
Two blind
One exophthalmic
Two exophthalmic
One hemorrhagic
Two hemorrhagic
One missing
Two missing
Other
Gills
Normal
Frayed
Clubbed
Marginate
Pale
Other
Pseudobranchs
Normal
Swol1 en
Lithic
Swollen & lithic
Inflamed
Other
B
R
G
NO
E
OT
0
1
2
H
S
M
G
U
OT
A
B
C
D
E
F
OT
Spleen
Black
Red
Granular
Modular
Enlarged
Other
Hind Gut
No inflammation
Mild inflammation
Severe inflammation
K1dneys
Normal
Swol1 en
Mottled
Granular
Urolithiasis
Other
Li yer
Normal, red
Pale red
Fatty
Nodules
Focal discoloration
General discoloration
Other
284
-------
ENTRY REQUIREMENTS (CONTINUED)
'ENTRY
0
1
2
0
1
2
3
4
0
1
2
EXPLANATION
Thyraus
No hemorrhage
Mild hemorrhage
Severe hemorrhage
Mesenterjc Fat
None
Little; <50% coverage
10% coverage
>50% coverage
100%
Fins
Normal
Mild active erosion
Severe actiave erosion
ENTRY
0
1
2
3
M
F
U
0
1
2
EXPLANATION
Bile
Yellow bile;
-------
10.10.2 Sample Report (Summary of Necropsy).
LOCATION:
Green River
QUALITY CONTROL NO.: 88-238
Autopsy Date: 7-6-
Age: 14 mos
Species: CUT
Strain: Bear Lake
Mark/Lot: 15Z5
Unit: Little Hole Water Temp.: 50
Fish Source: Whiterocks Water Hardness:
Sample Size
Tissue Collection No.
Disease Survey No.
F Case History No.
260 ppm Custody No.
Egg Source: Egan Investigator: Barton, Purpose Code
Hatching Date: 4-23-87 Reason for Autopsy: Green River Project
Remarks: Plasma samples: A403 to 414
60
NA
NA
NA
NA
0
Length
Weight
Ktl*
Ct]**
Hematocrit
Leucocrit
Plasma Protein
MEAN
222.330 nro
117.820 gr
1.070
3. 856
40.710
1.690
6.660
STANDARD
DEVIATION
20.69 rnn
39.81 gr
0.94
4.69
0.51
0.72
COEFFICIENT
OF VARIATION
9%
34%
88%
12%
30%
11%
*Expressed as Ktl times 10 to the fifth power
**Converted from Ktl; expressed as Ctl times 10 to the fourth power
VALUES AS PERCENT OF TOTAL SAMPLE
EYES
N 100%
81 0%
82 0%
El 07.
E2 0%
HI 0%
H2 0%
Ml 0%
M2 0%
OT 0%
SILLS
N 100%
F 0%
C 0%
M 0%
P 0%
OT 0%
PSEUDQ-
BRANCHS
N 1007.
S 0%
L 0*
S&L 0%
I 0%
OT 0%
THYMUS
0 43%
1 52%
2 5%
x 0.6
MESEN.
FAT
0 20%
1 40%
2 7%
3 25%
4 8%
x 1.6
SPLEEN
B 27%
R 73%
Q 0%
NO 0%
E 0%
OT 0%
HIND
GUT
0 83%
1 17%
2 0%
x 0.2
KIDNEY
N 100%
S 0%
M 0%
S 0%
U 0%
OT 0%
LIVER
A 12%
B 88%
C 0%
D 0%
E 0%
F 0%
OT 0%
BILE
0 63%
1 30%
2 7%
3 0%
x 0.4
FIN
0 47%
1 35%
2 18%
x 0.7
OPERCLE
-0 77%
1 13%
2 10%
x 0.3
100%
100%
100%
Summary of Normal:
43% xxxxxxx
100%
83%
100% 100% xxxxxxx
47%
77;
xxxxxxx i xxxxxxx
xxxxxxxx
Summary of Means
0.6
1.6
xxxxxxx 0.2
xxxxxxx
xxxxxx
0.4
0.7
0.3
SEX: M: 62% F: 38% U: 0%
Index Summary
Fat Index:
Bile Index:
Thymus i ndex
1.62
0.43
Gut Index,
Opercle Index
Fin Index
8.3
16.7
35.8
Normality Index
Severity Index
85.0
22.9
GENERAL REMARKS
FINS Left pelvic fin clipped;
SKIN Red dye marked
GONADS NA
OTHER 3 fish w/rttild infTarnation of hind gut
286
-------
10.10.2 Sample Report (Summary of Necropsy ) Continued.
Qua!. Control No. 88-23B
SN L6H «6T Ktl EYE SILL PSBR THY FAT SPL GUT KID LIV BILE SEX HEM LEU PLPR FIN OPCL
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
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
209
220
195
207
210
214
221
210
219
215
195
195
226
230
222
223
205
208
230
203
218
235
233
238
232
270
255
225
226
251
232
220
217
227
209
230
217
207
205
220
236
215
232
247
232
227
236
218
234
241
241
234
250
210
220
215
250
223
115
240
74 0,81
90 0,85
68 0-92
81 0.91
79 0.85
86 0.88
89 0.82
85 0.92
85 0.81
82 0.83
60 0.81
63 0.85
111 0.96
99 0.81
98 0.90
102 0.92
70 0.81
69 0.77
116 0.95
75 0.90
89 0.86
114 0.88
116 0.92
121 0.90
108 0.86
186 0.94
136 0.82
99 0.87
105 0.91
151 0.95
112 0.90
93 0.87
82 0.80
101 0.86
81 0.89
115 0.95
91 0.89
78 0.88
75 0.87
90 0.85
187 1.42
128 1.29
153 1.23
200 1.33
169 1.35
153 1.31
200 1.52
169 1.63
153 1.19
172 1.23
133 0.95
164 1.28
210 1.34
175 1.89
162 1.52
162 1.63
107 0.68
141 1.27
123 8.09
171 1.24
N
H
N
N
N
N
N
N
N
N
N
N
N
H
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
K
N
N
N
N
N
N
N
H
K
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
H
N
H
n
N
N
N
N
N
N
N
K
N
N
N
N
N
N
H
N
N
N
1
0
0
1
1
0
1
1
1
0
0
0
1
0
1
1
0
1
1
0
0
0
0
2
0
1
0
1
1
1
0
1
1
1
0
0
1
1
0
1
0
1
1
2
0
1
0
1
1
1
0
1
1
2
0
1
1
0
0
0
0
0
1
1
0
1
1
1
1
1
0
0
1
1
1
1
1
0
1
3
0
1
1
1
0
2
0
1
1
2
2
1
1
1
1
1
0
1
0
0
4
3
3
3
3
3
3
3
3
4
4
2
3
3
3
3
3
3
4
4
R
R
B
B
R
R
R
R
R
R
R
R
R
R
R
R
B
B
R
R
R
B
R
B
B
R
R
R
R
R
B
R
R
R
R
R
B
R
R
R
R
R
R
B
B
B
R
R
R
R
B
R
R
R
B
R
B
R
R
B
0
0
0
1
0
0
0
0
0
0
0
0
0
0
0
1
0
1
0
1
0
0
1
0
0
0
0
0
0
0
0
1
0
0
1
0
0
0
0
1
1
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
K
N
N
N
N
N
N
N
B
B
B
B
B
B
B
B
B
B
B
B
B
B
B
B
B
B
B
A
B
B
B
B
B
B
B
B
B
B
B
B
B
B
B
B
B
B
B
B
B
A
B
B
A
B
B
B
B
A
B
B
B
B
B
A
B
8
A
A
0
0
1
1
0
0
0
1
1
0
0
0
0
1
1
1
0
0
1
1
0
0
0
1
0
0
0
g
0
0
0
1
0
0
1
0
0
0
0
1
0
0
2
2
0
1
1
1
2
0
0
0
2
0
0
0
0
0
1
1
F
M
F
M
H
M
H
M
F
F
M
M
M
M
F
F
M
F
F
M
M
F
M
M
F
M
F
F
F
M
M
F
M
H
F
M
F
F
M
F
M
H
F
H
F
M
F
M
H
M
M
M
M
M
F
M
M
M
F
M
38
44
42
46
42
40
41
38
42
45
41.5
37
38
41
36
40
52
47
36
41
37
38
34.5
36
33
42
42.5
36.5
40
35.5
38
35
37
37
37.5
33
34
34
41
41
46
49
41
41
49
46
40.5
44
42,5
45
39
41
30
44
43.5
46
50
49
45.5
1
1.5
1.5
1
1.5
1.5
2
2
2
1.5
2
2
2.5
2
2
1
1.5
1
1.5
2
2.5
2.0
2.5
2.0
1.5
2,0
2.0
2.5
2.5
2.0
2.0
2.0
2.0
1.5
2.5
1.5
2.0
2.0
1.5
2.0
0.5
2.0
1.0
1.0
1.0
1,0
0.5
1.0
1.5
2,0
2
2
1.5
1
1
1.5
2
2.5
1.5
6.8
7.1
6,1
7.3
6.0
6.5
7.0
6,8
6.1
6.4
5.7
7.1
6.6
5.8
6,0
7.0
6.9
6.1
6.0
6.7
6.3
6.6
6.2
6.3
6.1
6.0
6.5
6.4
7.0
6.7
6.0
5.9
5.5
6.5
7.1
5.0
6.5
7.1
6.7
6.3
7.9
7.1
6.8
7.0
9.4
8.1
8.1
7.0
7.2
7.1
6.7
6.3
5
7.3
7
7.1
7.4
7.1
6.8
2
2
1
0
2
0
1
0
2
1
1
0
1
0
0
2
2
0
2
2
0
1
0
1
1
2
1
1
0
1
1
2
1
1
1
1
2
1
1
1
0
1
0
0
0
0
0
0
0
0
0
0
0
0
0
1
0
0
0
0
1
1
1
1
1
2
2
2
2
1
2
1
2
1
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
0
0
0
287
-------
10.11 Literature Cited
Goede, R. W. 1988. Fish health/condition assessment procedures. Part 2, A
color atlas of autopsy classification categories. Utah Division of
Wildlife Resources, 1465 West 200 North, Logan, UT 84321.
Goede, R. W. 1992. Fish health/condition assessment procedures. Part 1.
Utah Division of Wildlife Resources, Fisheries Experiment Station, 1465
West 200 North, Logan, UT 84321. 30 pp.
Goede, R. W. and B. A. Barton. 1990. Organismic indices and an autopsy-based
assessment as indicators of health and condition of fish. In: S, M.
Adams (ed.). Biological indicators of stress in fish. American
Fisheries Symposium 8, American Fisheries Society, Bethesda, Maryland.
pp. 93-108.
Goede, R. W. and S. Houghton. 1987. AUSUM A computer program for the
autopsy-based fish health/condition assessment system. Utah Division of
Wildlife Resources, Fisheries Experiment Sation, Logan, Utah 84321.
Lagler, K.F. 1962. Atlas of fish anatomy. Plate I, IV, and V. Michigan
Fisheries No. 5, Department of Fisheries, School of Natural Resources,
The University of Michigan, Ann Arbor, MI.
Novotny, J.F. and J. W. Beeman. 1990. Use of a fish health condition profile
in assessing the health and condition of juvenile chinook salmon.
Progr. Fish-Cult. 52:162-170.
288
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SECTION 11
GUIDELINES FOR FISH SANPLING AND TISSUE PREPARATION
FOR BIOACCUHULATIVE CONTAMINANTS
11.1 Introduction
11.1.1 Sampling of fish and shellfish for bioaccumulative contaminants has
been conducted for over 35 years. Most fish sampling for contaminants has
focused on contaminants of local concern, so data results and program
conclusions have not always been comparable. The issues surrounding
management of chemical contaminants in fish are of increasing concern for
fishery management, environmental and public health agencies. The
interdisciplinary multiagency problems caused by chemical contaminants
suggests the need for standard sampling protocols. There have been
inconsistent warnings given to the public by local, state, and federal
regulatory agencies regarding the consumption of sport fish. This has been
particularly evident on bodies of water shared by two or more states and on
international waters. The Great Lakes States (Great Lakes Fish Consumption
Advisory Task Force) and those States and EPA Regions bordering the
Mississippi (Mid-America Fish Contaminants Group) and Ohio Rivers (Ohio River
Valley Water Sanitation Commission) have endeavored to provide consistent
sampling and advisory information but a standard protocol has yet to be agreed
upon.
11.1.2 The application of quantitative risk assessment including hazard
assessment, dose response assessment, exposure assessment and risk
characterization functions best with a standardized protocol. The development
of human health fish consumption advisories, whether based on quantitative
risk assessment or some other methodology, is fundamentally affected by the
procedures used in sampling. This section presents guidance for the sampling
and preparation of fish for contaminant analysis, which is a key component of
exposure assessment in quantitative risk assessment.
11.1.3 The purpose and goals of each study should be clearly stated prior to
the initiation of fish collection for contaminant analysis. One should
consider the overall long-term development of a fish contaminant database in
each jurisdiction. Frequently short term goals have been the only
consideration, where as long term trend assessments may provide a better
understanding of the problem because the long, view is the only way of gauging
important changes occurring in water quality.
11.1.4 Various federal, state, and local agencies have responsibilities for
the collection and preparation of fish samples. Thus, numerous collection
protocols are available. Fish sampling for contaminant analysis will often be
included in other biological surveys to maximize use of the resource and to
minimize costs. It must be recognized that any sample collected represents
the future expenditure of significant dollar amounts by the time a decision is
reached, and can have significant effects on major sectors of our society.
289
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11.1.5 These guidelines present a basic fish sampling protocol designed to
give comparable results between studies. Some additional requirements are
pointed out which may be needed in special studies where different sizes or
species of fish might be targeted or where special collections for spike
samples might be needed. A partial discussion of sampling strategy including
statistical concerns can be found in USEPA (1989), which should be reviewed
during any planning effort.
11.2 Site Selection
11.2.1 Collecting sites should be established according to the specific
requirements of each study. Sites may be designed as short- or long-term
depending on the frequency with which they are sampled. Most sampling designs
for short-term (synoptic) studies will be structured to determine the extent
of contamination in a water body or a section of a water body. The
determination of contamination gradients extending away from point sources or
industrial/urban areas with point and non-point sources provides important
information needed to manage contaminant burdens in fish. Some sites will be
selected by individual states to address intrastate needs while other sites
will be selected to address interstate needs through cooperative programs.
Regardless of the various reasons for site selection, long-term comparability
is of utmost importance to provide trend information needed to place
bioaccumualtive contaminants in perspective.
11.2.2 Sites should be described as sport, commercial, or having both types
of fisheries, and additional sites may be identified for ecological risk
assessment. Special watershed information should be indicated, including
urban areas, mining, manufacturing, agriculture, etc., and any known point or
non-point sources of pollution at or near the site in the watershed.
Additional information should include average width, depth, and velocity at
the sampling station, description of the substrate, duration of the sampling
effort, and habitat area sampled (e.g., length of stream or area of lake).
Selected water quality measurements (e.g., conductivity, pH, dissolved oxygen,
temperature, etc.) may also be useful. It is becoming routine to collect and
analyze water, sediment and fish at common stations to gain a more complete
understanding of contaminants in aquatic environments.
11,3 Sample Collection
11.3.1 The following three objectives should guide sample collection:
1. Provide comparable data
2. Utilize sizes and ages of species generally available to
the fishery and,
3. Yield data which will screen for problems that might
indicate that more intensive studies are needed.
11.3.2 Samples should be obtained at each station from the principal fish
categories. Fish species are grouped by feeding strategy into predators,
omnivores and bottom feeders. To reduce the number of categories, the
290
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omnivores may be placed with the bottom feeders. USEPA (1990a) sampled 388
sites nationwide at which 119 different species of fish representing 33
taxonomic families of fish were collected. The most frequently sampled
freshwater and marine species in that study are listed in Table 1.
11.3.3 This national study indicates that of the freshwater species, carp and
largemouth bass were the most frequently sampled and are the most likely to
provide interstate comparability. The other freshwater species listed may be
selected in a declining order of priority; however, additional less common
species may not be added except in special situations. The diversity of
marine species is much greater resulting in a lack of focus on a limited
number. Additional effort will be needed to determine which marine species
should receive priority on the Atlantic, Pacific and Gulf Coasts in order to
provide long term comparative data.
11.3.4 Cunningham et al. (1990) in a census of state fish/shellfish
consumption advisory programs found that approximately 60 species of fish and
shellfish are used as the basis for consumption advisories nationwide. The
leading fish families are the Ictaluridae (catfish), Centrarchidae (sunfish,
largemouth and smallmouth bass), Cyprinidae (carp), and Salmonidae (salmon and
trout). Among shellfish, crustaceans (e.g., blue crab) and molluscs (e.g.,
American oyster, soft-shelled clam, and blue mussel) are the most widely used.
The criteria most frequently used for collecting fish/shellfish species were:
1) the dominant species harvested for consumption, 2) the most abundant
species and 3) the species representing a specific trophic order.
11.3.5 Consistent sampling of common species over long time periods (several
years) and large geographic areas will greatly facilitate future trend
analyses. Many species are similar in appearance, and taxonomic
identification must be reliable to prevent mixing species. Under no
circumstance should two or more species be mixed to create a composite sample.
Fish for contaminant analyses may be obtained during studies to determine fish
community structure. The measurement of multiple parameters (e.g., fish
health condition assessment, histopathological examination, bioindicators of
stress, etc.) are encouraged on common samples to provide the information
needed in ecological risk assessment,
11.3.6 Screening studies should endeavor to collect the largest individuals
available. However, more detailed studies should sample the predominant two
or three age classes of the same species in a water body to determine the
relationship between contaminant burden and fish size (age) to provide
information needed for greater risk management flexibility. This information
could allow the lifting of an advisory on smaller, more abundant sizes of a
contaminated species with lower body burdens if these were important to a
sport fishery.
11.3,7 The frequency of sampling should be considered in each study design.
Most long-term monitoring programs will be based on an annual frequency due to
the costs of analysis. However, special studies may require seasonal
sampling. Fish sampled in the fall may tend to have a higher lipid content
than those sampled during the spring. Sampling freshwater in the spring may
291
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TABLE 1. FREQUENCY OF OCCURRENCE FOR FRESHWATER AND MARINE SPECIES
STUDY (USEPA, 1990a>
IN THE NATIONAL FISH BIQACCUMULATION
FRESHWATER
Bottom Feeder Specieg
Carp
White sucker
Channel Catfish
Redhorse sucker
Spotted sucker
Site Occurrence
135
32
30
16
10
Game(Predator)Species
Largemouth Bass
Smallmouth Bass
Walleye
Brown trout
White Bass
Northern Pike
Flathead Catfish
White Crappie
Rainbow trout
Site Occurrence
83
26
22
10
10
8
8
7
7
MARINE
Hardhead catfish
Starry flounder
Blue fish
White perch
Winter flounder
White sturgeon
Red drum
Black drum
Striped mullet
Atlantic croaker .
Spot
Spotted seatrout
Weakfish
Sheepshead
Southern flounder
Flathead sole
Atlantic salmon
Red snapper
Gizzard shad
Atlantic cod
Yellow jack
Striped bass
American shad
Surf smelt
Spotted drum
Crevalle jack
Redstripe rockfish
Summer flounder
Diamond turbot
Hornyhead turbot
Bocaccio
White surfperch
Quillback rockfish
Brown rockfish
Copper rockfish
American eel
Site Occurence
7
5
5
4
4
4
3
3
3
3
3
3
3
2
2
2
2
2
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
292
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find fish more available due to spawning movements exhibited by spring
spawning species; however, extensive movement may temporarily dislocate fish
from the usual area where they have been exposed to contaminants. The various
methods of collecting fillets (skin-on versus skin-off, belly flap included or
excluded) must be standardized. A skin-on fillet with belly flap included is
recommended. A lipid analysis of each sample is required for trend analysis
and model validation, however, lipid content is not recommended for use in
normalizing the differences among fillet types because it frequently increases
the variance in the data (NOAA, 1989), Even when considering the
bioaccumulation of lipophilic compounds all of the compound is typically not
stored in the lipid. At any given time additional amounts of the compound
will be found in the cell moisture and the non-1ipid tissue. Lipid content
may also provide insight into seasonal changes within species, as well as
identify differences between species used in contaminants monitoring.
11.3.8 Active sampling techniques (electrofishing, trawling, seining, etc.)
are preferred over passive capture techniques (gill nets, trammel nets, etc.)
however, the latter can be used as long as the gear is checked on a frequent
basis to avoid sample deterioration. Species that are difficult to collect
may be obtained from a commercial fisherman, but only when the collector
accompanies the fisherman to verify the time and place of capture. Following
collection, fish should be placed on wet ice in clean coolers prior to
processing. Fish should be either processed within 24 hours or frozen within
24 hours for later processing if immediate processing is not possible. If
analyses of fish eggs or internal organs are required, a sample size of at
least 20 grams is required.
11.3.9 Composite samples of three to ten fish (same species) are recommended
for each of the predator and bottom feeder categories based on the variability
of contaminant concentration in fish at the site. The number of
fish/composite selected should remain constant over time and space for each
species monitored. Composites are used to reduce the cost of analysis per
fish; however, it must be recognized that statistical manipulation of the data
is compromised when individual values are not determined. The smallest size
fish in a composite should equal 75% of the total length of the largest fish
in a composite, e.g., if the largest is 400 mm, the smallest should not be
less than 300 mm. Replicate composite samples may be added as needed to meet
statistical requirements; (USEPA, 1989) however, the cost of additional
samples will quickly become a factor. The most important sport and/or
commercial species in each feeding strategy group should be used for analysis.
Composite samples can be collected for either fillet analysis (human health
risk assessments) or for whole body analysis (ecological risk assessments and
worst case monitoring).
11.3.10 When a study is planned, it is not certain that the quantity of each
species indicated for analysis can be obtained especially if the water body
has had little or no prior sampling activity. In order to meet both the human
health and ecological requirements a sample of a sport fish species and a
bottom feeder species is needed. The sport fish species is usually filleted
and the data used for human health risk assessment. The whole body analysis
of bottom feeder species is used both for initial "worst case" monitoring and
for ecological risk assessment.
293
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11.3.11 If fish are not abundant or detailed comparisons with other
parameters are desired, it may be possible to do a reconstructed analysis
(Figure 1) on a single species either sport fish or bottom feeder. To do a
reconstructed analysis, the fish are filleted and the remainder of the carcass
is saved for analysis. The contaminant concentrations in both the fillet and
remaining carcass portions can then be added together to estimate the whole
body concentration. A lipid analysis must be performed on both the fillet and
remaining carcass to allow normalization of the contaminant concentrations in
both samples. A reconstructed analysis may be performed on either single fish
or composite fish samples, however, the data may be more reliable if single
fish are analyzed.
11.3.12 Sediment samples can sometimes indicate a "hot spot" and can be
helpful in determining the source(s) of contamination or the zones of
deposition. However, sediment samples cannot be used as a substitute for fish
collections, but both can provide complimentary data.
11.4. Sample Preparation For Organic Contaminants in Tissue
11.4.1 Collection Precautions
11.4.1.1 In the field, sources of tissue contamination include sampling gear,
boats and motors, grease from ship winches or cables, engine exhaust, dust,
and ice used for cooling. Efforts should be made to minimize handling and to
avoid sources of contamination. For example, to avoid contamination from ice,
the whole samples (e.g., molluscs in shell, whole fish) should be wrapped in
aluminum foil, placed in watertight plastic bags, and immediately cooled in a
covered ice chest. Many sources of contamination can be avoided by resecting
(i.e., surgically removing) tissue in a controlled environment (e.g., a
laboratory). Organisms should not be frozen prior to resection if analyses
will be conducted on only selected tissues (e.g., internal organs) because
freezing may cause internal organs to rupture and contaminate other tissue.
If organisms are eviscerated in the field, the remaining tissue may be wrapped
as described above and frozen. Tissue sample collection and preparation
requirements are summarized in Table 2 (Puget Sound Estuary Program, 1989).
11.4.2 Processing
11.4.2.1 To avoid cross-contamination, all equipment used in sample handling
should be thoroughly cleaned before each sample is processed. All instruments
must be of a material that can be easily cleaned (e.g., stainless steel,
anodized aluminum, or borosilicate glass). Before the next sample is
processed, instruments should be washed with a detergent solution, rinsed with
tap water, rinsed in isopropanol, and finally rinsed with organic free
distilled water. Work surfaces should be cleaned with isopropanol, washed
with distilled water and allowed to dry completely.
/
11.4.2.2 The removal of biological tissues should be carried out by or under
the supervision of an experienced biologist. Tissue should be removed with
clean stainless steel or quartz instruments (except for external surfaces).
The specimens should come into contact with precleaned glass surfaces only.
Polypropylene and polyethylene (plastic) surfaces and implements are a
294
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Sample
Station
Hunan
Receptor
to
tn
Animal
Receptor
Insufficient Quantities
of Sport Fish or Bottom
Feeder Species are Found
Sufficient
Quantities
of Sport Fish
Found
Sufficient
Quantities
of Bottom Feeders
Found
Replicate
Compos i te
Fillet
Sample #3
Perform
Reconstructed
Analysis
Sample #1
Figure 1. General sampling scheme for bioaccumulative contaminants in fish, multiple age groups will require additional
samples.
-------
TABLE 2. SUMMARY OF SAMPLE COLLECTION AND PREPARATION QA/QC REQUIREMENTS FOR FISH
TISSUE (MODIFIED FROM PUGET SOUND ESTUARY PROGRAM, 1986, 1989)
Variable Samp!e Size (a) Cont.ajner (b) Preservation
Organic Compounds
Who]ebody li s s ues
(after resection)
Semivolatiles
Volatiles
25 g
5 9
G,T,A
G,T
Maximum Hoiding
Time (c)
Freeze (-18°C) 1 yr
Freeze (d) (-18°C) 1 yr
Freeze (d) (-18°C) 14 days
Maximum Extract
Holding Time
40 days
40 days
Trace Metals
Wholebody Tissues
W,P,B
Freeze
6 mo
(after resection)
All Metals
(except Hg)
5 g
0.2 g
P,B Freeze (d)
P,B Freeze (d)
6 mo
28 days
a. Recommended wet weight sample sizes for one laboratory analysis. If additional laboratory
analyses are required (i.e., replicates) the field sample size should be adjusted
accordingly. If specific organs are to be analyzed, more tissue may be required.
b. G = glass, A = wrapped in aluminum foil, placed in watertight plastic bags, T = PTFE
(Teflon), P = linear polyethylene, B = borosilicate glass, W = watertight plastic bags.
c. This is a suggested holding time. No USEPA criteria exist for the preservation of this
variable.
d. Post-dissection
-------
potential source of contamination and should not be used. To control
contamination when resecting tissue, technicians should use separate sets of
utensils for removing outer tissue and for resecting tissue for analysis.
11.4.3 Preparation of Composite Fillet Samples
11.4.3.1 For fish samples, special care must be taken to avoid contaminating
targeted tissues (especially muscle) with slime and/or adhering sediment from
the fish exterior (skin) during resection. The proper handling in the
preparation of fish tissue samples to decrease the likelihood of contamination
cannot be over emphasized. To reduce variation in sample preparation and
handling, samples should be prepared in the laboratory rather than in the
field. However, if no laboratory is available, field preparation is
acceptable if portable tables are used, dust and exhausts are avoided and
proper decontamination procedures are followed. Regardless of where
preparation occurs, the following subsections should be followed to insure
quality fillet samples:
11.4.3.2 To initiate processing, each fish is measured (total or fork length)
to the nearest tenth of a centimeter, weighed (nearest gram) and external
condition noted. A few scales should be removed from each fish for age and
growth analysis. This presents an excellent opportunity to systematically
evaluate each fish using the Fish Health and Condition Assessment Methods
(Section 10). Fish are scaled (or skinned: catfish) and filleted carefully,
removing bones, to get all of the edible portion flesh.
11.4,3.3 A fillet includes the flesh tissue and skin from head to tail
beginning at the mid-dorsal line from the left side of each fish and including
the belly flap. The fillet should not be trimmed to remove fatty tissue along
the lateral line or belly flap. A comparable fillet can be obtained from the
right side of the fish and can be composited with the left fillet, kept
separate for duplicate quality assurance analysis, analyzed for different
compounds or archived. Each right and left fillet should be weighed
individually, recorded and individually wrapped in clean aluminum foil.
11.4.3.4 Care must be exercised not to puncture any of the internal organs.
If the body cavity is entered, rinse the fillet with distilled water. Fish
sex and condition of internal organs are determined during or after filleting.
This skin-on fillet deviates from the skin-off fillets analyzed in the
National Fish Bioaccumulation Study (USEPA 1990a), however, skin-on is
recommended because it is believed that this is the way most sport anglers
prepare their fillets. The issue of skin-on versus skin-off fillets differs
greatly among jurisdictions (Hesse, 1990) and is far from settled, however,
the above recommendations appear to be the preferred method unless the species
specificity is increased in future guidelines.
11.4,3.5 Filleting should be conducted on cutting boards covered with heavy
duty aluminum foil, which is changed between composite samples. Knives, fish
sealers, measurement boards, scales, etc. should be cleaned with reagent
grade isopropanol, followed by a rinse with distilled water between each
composite sample.
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11.4.3.6 Because of the low limits of detection for many environmental
analyses, clean field and laboratory procedures are especially important.
Sample contamination can occur during any stage of collection, handling,
storage or analyses. Potential contaminant sources must be known and steps
taken to minimize or eliminate them.
11.4.3.7 Large sheets of heavy duty aluminum foil should be used to carefully
fold and completely wrap the fillet samples. When filling out I.D. labels use
pencil or waterproof marker and place the foil wrapped sample in a secured
plastic bag.
11.4.4 Storage
11.4.4.1 Recommended holding times for frozen tissue samples have not been
established by USEPA, but a maximum 1 year holding time is suggested. For
extended sample storage, precautions should be taken to prevent desiccation.
National Institute For Standards and Technology is testing the effects of
long-term storage of tissues at temperatures of liquid nitrogen(-120° to -
190 C). At a minimum, the samples should be kept frozen at -20°C until
extraction. This will slow biological decomposition of the sample and
decrease loss of moisture. Liquid associated with the sample when thawed must
be maintained as part of the sample because the lipid tends to separate from
the tissue. Storage of samples should remain under the control of the sample
collector until relinquished to the analytical laboratory.
11,4.4.2 Whole fish may be frozen and stored if no resection of internal
organs or fillets will be conducted and the ultimate analysis is whole body.
However, if resection of fillets or organs is required, these tissues should
be removed prior to freezing and can be stored frozen in appropriate
individual containers. The tissues may then be ground and homogenized at a
later date and refrozen in sample packets for shipment on dry ice to the
analytical laboratory(s).
11.4.4.3 It is frequently necessary to ship whole fish, fillets or
homogenized tissue samples over long distances to an analytical laboratory.
To avoid sample deterioration, it is recommended that all samples be frozen
solid prior to shipment. The frozen and logged samples should be wrapped in
newspaper to provide additional insulation for the samples which are shipped
in well sealed insulated containers with an appropriate quantity of dry ice.
The quantity of dry ice should be sufficient to eliminate any defrosting of
the samples during the time of priority transport. However, in the event that
a delay occurs in transit, these recommendations will provide some assurance
that the samples will arrive in usable condition. Under no circumstances
should unfrozen tissue be shipped either with or without dry ice because the
quality of the sample cannot be assured.
11.4.5 Tissue Preparation
11.4.5.1 Organic contaminants are not evenly distributed throughout
biological tissue, especially in fish. This is also true for fish fillets.
Therefore, to obtain a homogenous sample, the whole fish or the whole fillet
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must be ground to a homogeneous consistency. This procedure should be carried
out by the sample collector on partially thawed samples,
11.4.5.2 Chop the sample into 2.5 cm cubes unless the sample is small enough
to fit in a hand crank meat grinder (300 gm or less) or a food processor
(Hobart Model 8181D or equivalent for large fish) (USEPA, 1990b). Then pass
the whole sample through a meat grinder. Grinding of biological tissue is
easier when the tissue is partially frozen. This is especially true when
attempting to grind the skin. Chilling the grinder with a few chips of dry
ice will reduce the tendency of the tissue to stick to the grinder. Do not
freeze the grinder since hard frozen tissue is difficult to force through the
chopper plate.
11.4.5.3 The ground sample is divided into quarters, opposite quarters are
mixed by hand with a clean stainless steel spatula and then the two halves are
mixed back together. Repeat the mechanical grinding, quartering and hand
mixing two more times. No chunks of tissue should be present at this point as
they will not be efficiently extracted. Very small fish or small fillets may
be homogenized in a high speed blender.
11.4.5.4 When compositing fillets or whole fish each individual fillet or
fish should be ground separately following the above described procedure.
Then take equal amounts from each fillet or fish sample to be composited to
provide a total equal to that required for extraction or the total number of
split and archived samples required by the study plan.
11.4,5.5 If the ground fish is to be re-frozen prior to extraction and
analysis, weigh out the exact amount for extraction into a small container.
Using a top loading balance, tare a 2 oz. glass jar (or a small sheet of
aluminum foil that can be formed into a sealed packet) to 0.0 gm and carefully
dispense a 20.0 gm portion of homogenized tissue into the container. Tightly
seal the container or foil packet. Repeat with additional containers for
duplicates, splits, or archived samples. Lipid material tends to migrate
during freezing; therefore, storing a weighed portion ensures extraction of a
representative portion of the tissue if the foil or container is completely
rinsed with solvent by the analytical chemist.
11.4.5.6 Whenever a ground sample is to be split between two or more labs,
the ground sample must also be mixed with reagent grade anhydrous sodium
sulfate (previously heated to 400°C to drive off any phthalate esters acquired
during storage). To ensure the homogeneity of the sample prior to splitting,
transfer 100 gm of ground tissue to a 600 ml beaker. Add 250 gm of anhydrous
sodium sulfate and mix thoroughly with a stainless steel spoon or a spatula.
There should not be any lumps and the mixture should appear homogeneous.
Dispense exactly 70.0 gm of mixture to each lab and note on the package that
it contains 20 gm of tissue.
11.4.5.7 When preparing the tissue for volatile analysis, grind it in an area
free of volatile organic compounds. The meat grinder or food processor must
be heated in an oven for 30 minutes at 105°C after solvent rinsing and then
allowed to cool at room temperature. Immediately after grinding the tissue,
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weigh duplicate 1 gm portions into culture tubes with screw caps. Analyze
immediately or store in a freezer,
11.5 Sample Preparation For Metal Contaminants In Tissue
11.5.1 Collection Precautions
11.5.1.1 The major difficulty in trace metal analyses of tissue samples is
controlling contamination of the sample after collection. In the field,
sources of contamination include sampling gear, grease from winches or cables,
engine exhaust, dust, or ice used for cooling. Care must be taken during
handling to avoid these and any other possible sources of contamination. For
example, during sampling the ship should be positioned such that the engine
exhausts do not fall on deck. To avoid contamination from melting ice, the
samples should be placed in watertight plastic bags.
11.5.1.2 Sample resection and any subsampling of the organisms should be
carried out in a controlled environment (e.g., dust-free room). In most
cases, this requires that the organisms be transported on ice to a laboratory
rather than being resected in the field. It is recommended that whole
organisms not be frozen prior to resection if analyses will be conducted only
on selected tissues, because freezing may cause internal organs to rupture and
contaminate other tissue. If organisms are eviscerated in the field, the
remaining tissue (e.g., muscle) may be wrapped as described above and frozen
(Puget Sound Estuary Program, 1986).
11.5.1.3 Resection is best performed under "clean room" conditions. The
"clean room" should have positive pressure and filtered air and also be
entirely metal-free and isolated from all samples high in contaminants (e.g.,
hazardous waste). At a minimum, care should be taken to avoid contamination
from dust, instruments, and all materials that may contact the samples. The
best equipment to use for trace metal analyses is made of quartz, TFE
(tetrafluoroethylene), polypropylene, or polyethylene. Stainless steel that
is resistant to corrosion may be used if necessary. Corrosion-resistant
stainless steel is not magnetic, and thus can be distinguished from other
stainless steels with a magnet. Stainless steel scalpels have been found not
to contaminate mussel samples (Stephenson et al., 1979). However, low
concentrations of heavy metals in other biological tissues (e.g., fish muscle)
may be contaminated significantly by any exposure to stainless steel. Quartz
utensils are ideal but expensive. To control contamination when resecting
tissue, separate sets of utensils should be used for removing outer tissue and
for removing tissue for analysis. For bench liners and bottles, borosilicate
glass would be preferred over plastic if trace organic analyses are to be
performed on the same sample.
11.5.1.4 Resection should be conducted by or under the supervision of a
competent biologist. Special care must be taken to avoid contaminating target
tissues (especially muscle) with slime and/or adhering sediment from the fish
exterior (skin) during resection. The procedure previously outlined for the
preparation of fillet samples should generally be followed. Unless
specifically sought as a sample, the dark muscle tissue that may exist in the
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vicinity of the lateral line should not be separated from the light muscle
tissue that constitutes the rest of the muscle tissue mass.
11.5.1.5 Prior to use, utensils and bottles should be thoroughly cleaned with
a detergent solution, rinsed with tap water, soaked in acid, and then rinsed
with metal-free water. For quartz, TFE, or glass containers, use 1+1 HN03,
1+1 HC1, or aqua regia (3 parts cone. HC1 + 1 part cone HN03) for soaking.
For plastic material, use 1+1 HNO, or 1+1 HC1. Reliable soaking conditions
are 24 h at 70°C (APHA, 1989; 1992). Do not use chromic acid for cleaning any
materials. Acids used should be at least reagent grade. For metal parts,
clean as stated for glass or plastic, except omit the acid soak step. If
trace organic analyses are to be performed on the same samples, final rinsing
with methylene chloride is acceptable.
11.5.1.6 Sample size requirements can vary with tissue type (e.g., liver or
muscle) and detection limit requirements. In general, a minimum sample size
of 6 g (wet weight) is required for the analysis of all priority pollutant
metals. To allow for duplicates, spikes, and required reanalysis, a sample
size of 50 g (wet weight) is recommended. Samples can be stored in glass,
TFE, or high-strength polyethylene jars.
11.5.2 Processing
11.5.2.1 Samples should be frozen after resection and kept at -2Q°C.
Although specific holding times have not been recommended by USEPA, a maximum
holding time of 6 months (except for mercury samples, which should be held a
maximum of 28 days) would be consistent with that for water samples.
11.5.2.2 When a sample is thawed, the associated liquid should be maintained
as a part of the sample. This liquid will contain lipid material. To avoid
loss of moisture from the sample, partially thawed samples should be
homogenized. Homogenizers used to grind the tissue should have tantalum or
titanium parts rather than stainless steel parts. Stainless steel blades used
during homogenization have been found to be a source of nickel and
chromium contamination. Some trace metal contamination during processing
cannot be avoided and it is therefore necessary to determine and control the
amount of contamination introduced during processing. Contamination can be
monitored by introducing a dry ice blank into the blender and analyzing the
chips.
11.5.2.3 To avoid trace metal contamination during processing the preferred
method is to proceed to a chemical digestion process which minimizes or
eliminates resection, homogenization, or grinding. Chemical digestion is best
limited to specific organ tissues from large fish or to smaller sized whole
fish.
11.6 Identification of Composite Whole Fish or Fillet Samples
11.6.1 Composite whole fish samples will be made up of three to ten fish with
any deviation in number clearly identified. The limitation on the variance
between individual fish in each composite will be as previously described.
The length and weight of each fish must be recorded. The same field
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information should be provided as described above for both fillet and/or whole
body composite samples. The same handling precautions as described above
should be followed for either organic or trace metal contaminants. Spines on
whole fish should be sheared to minimize puncturing the sample packaging.
11.6.2 The following information should be included on the field/lab form for
each sample collected:
11.6.2.1 Project Name
11.6.2.2 Station Code (if applicable)
11,6.2.3 Date
11.6.2.4 Collector's Name
11.6,2.5 Sampling location (river mile and/or other specific
information relating to local landmarks)
11.6.2.6 Latitude and Longitude
11.6.2.7 Water body name
11.6.2.8 Sampling technique(s), i.e. 230 vac electrofishing
apparatus, hoop nets, etc.
11.6.2.9 Fish species
11.6.2.10 Individual lengths and weights of fish in sample
11.6.2.11 Sample type (Whole or Fillet)
11.6.2.12 Individual fillet weights (whether left or right)
11.6.2.13 Comments or Unusual Conditions, i.e., tumors, sores,
fin rot, blind, etc.
11.7 Chain-of-Custody Procedures (USEPA, 1990c; USEPA, 1991)
Also See Section 2, Quality Assurance and Quality Control.
11.7.1 All samples should be kept in a secure (locked) area to avoid legal
complications in administrative proceedings. Transportation of the samples
must be coordinated between the agency responsible for the field collection
and the agency responsible for analytical work. When custody of the samples
is transferred, the following checks should be implemented:
11.7.1.1 All transfers should be properly relinquished to ensure ehain-of-
custody. Transfers should be recorded on a form separate from the field data
sheet. The chain-of-custody form should include the sample identification
number(s). Custody tags must be used and numbered in sequence (if possible).
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11.7.1.2 The field data sheet should stay with the sample until it is logged
in by the analytical laboratory.
11.7.1.3 Samples can be shipped and chain-of-custody maintained as long as
shipping containers are sealed with custody tape.
11.7.1.4 Samples should remain frozen until they are prepared for analysis.
Shipping with dry ice is recommended.
11.7.1.5 The laboratory's receiving agent should initial the field data sheet
and affix the date of sample receipt. Depending on administrative need, a
copy of this form (with initials and date of sample receipt plainly visible)
may be required by the lab agency's central office.
11.8 Conclusion
11.8.1 This protocol only addresses the steps to be considered in field
sampling fish and sample preparation for human health fish consumption
advisories and ecological risk assessment. Additional protocols must be
followed to carry out the appropriate analytical chemistry and the risk
assessment/management requirements leading to an action. These additional
protocols were beyond the scope of this assignment.
11.9 Literature Cited
Cunningham, P.A., J.M. McCarthy and D. Zeitlin 1990. Results of the 1989
Census of State Fish/Shellfish Consumption Advisory Programs. Prepared
for S.M. Kroner, Assessment and Watershed Protection Division, OWRS,
USEPA, by Research Triangle Institute, P.O. Box 12194, Research Triangle
Park, NC.
APHA. 1989. Standard methods for examination of Waste and Wastewater. 17TH
Ed. American Public Health Association, Washington, DC.
APHA. 1992, Standard methods for examination of Waste and Wastewater. 18TH
Ed. American Public Health Association, Washington, DC.
Hesse, John L. Michigan Department of Public Health, 1990. Summary and
Analysis of Existing Sportfish Consumption Advisory Programs in the
Great Lakes Basin. The Great Lakes Fish Consumption Advisory Task Force
Co-Chaired by H.A. Anderson and L. Liebenstein, State of Wisconsin.
Unpublished.
NOAA. 1989. A summary of data on tissue contamination from the first three
years (1986-89) of the mussel watch project. Technical Memorandum,
NOS, OMA49. Rockville, MD.
Puget Sound Estuary Program 1986. Recommended Protocols for Measuring Metals
in Puget Sound Water, Sediment and Tissue Samples. Prepared by Tetra
Tech, Inc., Bellevue, WA. In: Recommended Protocols for Measuring
Selected Environmental Variables in Puget Sound. USEPA, Region 10,
Seattle, WA (Looseleaf).
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Puget Sound Estuary Program 1989 (Revised). Recommended Guidelines for
Measuring Organic Compounds in Puget Sound Sediment and Tissue Samples.
Prepared by Tetra Tech, Inc., Bellevue, WA. In: Recommended Protocols
for Measuring Selected Environmental Variables in Puget Sound. USEPA,
Region 10, Seattle, WA (Looseleaf).
Stephenson, M.D., M. Martin, S.E. Lange, A.R. Flegal and J.H. Martin 1979.
California Mussel Watch 1977-78. Volume II: Trace metals
concentrations in the California mussel, Mytilus call form'anus. SWRCB
Water Quality Monitoring Report No. 79-22. Sacramento, CA.
USEPA, 1989. Assessing Human Health Risks from Chemically Contaminated Fish
and Shellfish: A Guidance Manual. EPA-503/8-89-002. Office of Marine
and Estuarine Protection and Office of Water Regulations and Standards,
Washington, DC.
USEPA, 1990a. Bioaccumulation of Selected Pollutants in Fish, A National
Study Volume I and II. EPA-506/6-9Q/001. Office of Water Regulations
and Standards (WH-552), Washington, DC.
USEPA, 1990b. Extraction and Analysis of Organics in Biological Tissue,
Method OB 8/90, USEPA, Environmental Services Division, Region IV,
Analytical Support Branch, Athens, GA.
USEPA, 1990c. Manual for the certification of laboratories analyzing drinking
water. Criteria and procedures quality assurance. EPA/570/9-90/008.
Prepared by the Laboratory Certification Program Revision Committee.
Office of Water (WH-550D), Washington, DC.
USEPA, 1991. Manual for the evaluation of laboratories performing aquatic
toxicity tests. EPA/600/4-90/031. Klemm, D.J., L.B. Lobring, and W.H.
Horning, II. Environmental Monitoring Systems Laboratory, U.S.
Environmental Protection Agency, Cincinnati, OH.
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SECTION 12
FISHERIES BIBLIOGRAPHY
12.1 General References
Adams, S.M, (ed.), 1990. Biological indicators of stress in fish. American
Fisheries Society Symposium 8, Bethesda, MD.
Alabaster, J.S. 1985. Habitat modification and freshwater fisheries.
Butterworth Pub!., Stoneham, MA.
Alabaster, J.S. and R. Lloyd. 1980. Water quality criteria for freshwater
fish. FAO, United Nations, Butterworths, Boston, MA.
Allen, G.H., A.C. Oelacy, and D.W. Gotshall. 1960. Quantitative sampling of
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APHA. 1992. Standard methods for the examination of water and wastewater.
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Angermeier, P.L. and R.J. Neves. 1991. Assessing stream values:
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Backiel, T. and R.L. Welcomme. 1980. Guidelines for sampling fish in inland
waters. FAO Technical Paper 33, UNIPUB, New York, NY.
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coasts. Cambridge Univ. Press, New York, NY.
Ballentine, R.K. and L.J. Guarrie (eds.). 1975. The integrity of water: a
symposium. U.S. Environmental Protection Agency, Washington, DC.
Barnes, R.S.K. and R.N. Hughes. 1982. An introduction to marine ecology.
Blackwell Scientific Publications, Ltd., Oxford, England.
Barnes, R.S.K. and K.H. Mann. 1991 (eds.). Fundamentals of aquatic ecology.
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Bell, M.C. 1986. Fisheries handbook of engineering requirements and
biological criteria. U.S. Army Corps Engineers, Portland OR.
Backiel, T. and R.L. Welcomme. 1980. Guidelines for sampling fish in inland
waters. Unipub. New York, NY.
Banarescu, P. (ed.). 1990. Distribution and dispersal of freshwater animals
in North America and Eurasia. Vol. 2. AULA-Verlag, Wiesbaden, Germany.
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Barnes, R.S.K. and R,N. Hughes, 1982, An introduction of marine ecology.
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Berkman, H.E. and C.F. Rabeni. 1987. Effect of siltation on stream fish
communities. Env. Biol. Fishes. 18:285-294.
Beitinger, T.L. 1990. Behavioral reactions for the assessment of stress in
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Bone, Q. and N.B. Marshall. 1982. Biology of fishes. Methunen, Inc.. Amer.
Fish. Soc., Bethesda, MD.
Bovee, K.D. 1982. A guide to stream habitat analysis using the instream flow
incremental methodology, U.S. Fish and Wildlife Service Biological
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Bramblet, R.G. and K. D. Fausch. 1991. Variable fish communities and the
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Bramblett, R.G. and K.D. Fausch. 1991. Variable fish communities and the
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Cailliet, 6.M., M.S. Love, and A.M. Ebeling. 1986. Fishes: a field and
laboratory manual on their structure, identification, and natural
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Carlander, K.D. 1969. Handbook for freshwater fishery Biology; life history
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Caulcutt, R. 1991. Statistics in research and development. Chapmann and
Hall, New York, NY.
Charles, D.F. (ed.). 1991. Acidic deposition and aquatic ecosystems:
Regional case studies. Springer-Verlag, New York, NY.
Cole, R.A. and J.P. Rockwood. 1989. Water pollution biology: A
laboratory/field handbook. Technomic Publishing Co., Inc., Lancaster,
PA.
Crossman, E.J. and J.M. Casselman. 1987. Pike bibliography. Royal Ontario
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Cummins. K.W. 1991. Establishing biological criteria: functional views of
biotic community organization. In: Biological criteria: research and
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Curtis, B. 1948. The life story of the fish. Harcourt, Brace and Company,
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Cushing, D.H. 1968. Fisheries biology. A study in population dynamics.
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Cushing, D.H. 1975. Marine ecology and fisheries, Cambridge Press,
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Cushing D.H. 1983. Key papers on fish populations. IRL Press, Oxford,
England.
Digby, P.G.N. and R.A. Kempton. 1987. Multivariate analysis of ecological
communities. Chapman and Hall, New York, NY.
DuBois, R.B. 1989. Bibliography of fishery investigation on large salmonid
river systems with special emphasis on the Bois Brule river, Douglas
County, Wisconsin. Technical Bulletin No. 166, Wisconsin Department of
Natural Resources, Madison, WI.
Duff, D.A. and N. Banks. 1988. Indexed bibliography on stream habitat
improvement. USDA, Forest Service, Intermountain Region, Ogden, UT.
Dumont, W.H. and G.T. Sundstrom. 1961. Commercial fishing gear of the United
States. Washington, DC, U.S. Government Printing Office, Fish and
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Edwards, E.F. and B.A. Megrey. 1989. Mathematical analysis of fish stock
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Everhart, W.H., A.M. Eipper, and W.D. Young. 1975. Principles of fishery
Science. Cornel Univ. Press, Ithaca, NY.
Evans, D.O., G.J. Warren, and V.W. Cairns. 1990. Assessment and management
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Fausch, K.D., J.R. Karr, and P.R. Yant. 1984. Regional application of an
index of biotic integrity based on stream fish communities. Trans.
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Fausch, K, D. Hawkes, and M. Parsons 1988. Models that predict standing
crop of stream fish from habitat variables: 1950-1985. Dept. Fishery
and Wildlife Biology, Colorado State University, Fort Collins CO.
Fausch, K.D., J. Lyons, J.R. Karr, and P. Angermeier. 1990. Fish communities
as indicators of environmental degradation. Amer. Fish. Soc. Symposium
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Freedman, B. 1989. Environmental ecology. Academic Press, Harcourt Brace
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Fridman. A.L. 1988. Calculations for fishing gear designs. Fishing News
Books Ltd., Farnham, Surrey, England.
Garner, J. 1988, Modern deep sea trawling gear. Fishing News Book Ltd.,
Farnham, Surrey, England.
Garner, J. 1989. Net work exercises. Fishing News Books Ltd., Farnham,
Surrey, England.
Gilbert, R.O. 1987. Statistical methods for environmental pollution
monitoring. Van Nostrand Reinhold Co., New York, NY.
Goldman, C.R. and A.J. Home. 1983. Limnology. McGraw-Hill, New York, NY.
Gonnason, L. 1989. Sonar for fisheries research: An introductory guide to
hydroacoustics. BioSonics, Inc., Seattle, WA.
Gorman, O.T. 1987. Habitat segregation in an assemblage of minnows in an
Ozark stream. In: W.J. Matthews and D.C. Heins (eds.). Community and
evolutionary ecology of North American stream fishes. Univ. Oklahoma
Press, Norman, OK.
Green, J. 1968. The biology of estuarine animals. Univ. Wash., Seattle, WA.
Grossman. G.D., P.B. Moyle, and J.O. Whitaker, Jr. 1982. Stochasticity in
structural and functional characteristics of an Indiana stream fish
assemblage: a test of community theory. Amer. Naturalist 120:423-454.
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Ming. A. 1964. Contributions to a bibliography on the construction,
development, use and effects of electrofishing devices. Okla. Fish.
Res. Lab., D-J Federal Aid Proj. FL-6, Semiann. Rept. (Jan-June, 1964).
pp. 33-46.
Monan, G.E. and D.E. Engstrom. 1962. Development of a mathematical relation-
ship between electra-field parameters and the electrical characteristics
of fish. U.S. Fish Wildl. Bull. 63:123-136.
Murray, A.R. 1958. A direct current electrofishing apparatus using separate
excitation. Can. Fish Cult. 23:27-32.
Northrop, R.B. 1962. Design of a pulsed DC-AC shocker. Conn. Bd. Fish and
Game, D-J Federal Aid Proj. F-25-R, Job No. 1.
Novotny, D.W. and G.R. Priegel. 1974. Electrofishing boats: Improved designs
and operational guidelines to increase the effectiveness of boom
shockers. Mis. Dept. Nat. Res., Tech. Bull. 73, Madison, WI. 48 pp.
Ohio EPA. 1989. Biological criteria for the protection of aquatic life:
Volume III. Standardized biological field sampling and laboratory
methods for assessing fish and macroinvertebrate communities. Ohio
Environmental Protection Agency, Columbus, OH.
320
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Omand, D.N. 1950, Electrical methods of fish collection. Can. Fish Cult.
9:13-20.
Petty, A.C. 1955. An alternate-polarity electrode. New York Fish Game J.
2:114-119.
Platts, W.S., M.F. Megahan, and G.W. Minshall. 1983. Methods for evaluating
stream riparian and biotic conditions. U.S. For. Serv. For. Range Exp.
Stn., Gen. Tech.Rep. INT-138. 70 pp.
Reynolds, J.B, 1983. Electrofishing. In: L.A. Nielsen and D.L. Johnson,
eds. Fisheries Techniques. Amer. Fish. Soc., Bethesda, MD. pp. 147-
164.
Reynolds, J.B. and D.E. Simpson. 1978. Evaluation of fish sampling methods
and rotenone census. In: G.D. Novinger and J.G. Dillard, eds. New
approaches to the management of small impoundments. North Central
Division, Amer. Fish., Special Pub!. 5:11-24.
Ruhr, C.E. 1953. The electric shocker in Tennessee. Tenn. Game Fish Comm.
(Mimeo). 12 pp.
Sanders, R.E. 1990. A 1989 night electrofishing survey of the Ohio river
mainstem (RM 280.8 to 442.5). Ohio Environmental Protection Agency,
Columbus, OH.
Sanders, R.E. 1992. Day versus night electrofishing catches from near-shore
waters of he Ohio and Muskingum Rivers. Ohio J. Sci. 92(3):In Press.
Saunders, J.W. and M.W. Smith. 1954. The effective use of a direct current
fish shocker in a Prince Edward Island stream. Can. Fish. Cult. 16:42-
49.
Schwartz, F.J. 1961. Effects of external forces on aquatic organisms.
Maryland Dept. Res. Edu., Chesapeake Biol. Lab., Contr. No. 168, pp. 3-
26.
Sharpe, F.P. 1964. An electrofishing boat with a variable-voltage pulsator
for lake and reservoir studies. U.S. Bur. Sport Fish, and Wild!.
Circular 195. 6 pp.
Sharpe, P.P., W.T. Burkhard. 1969. A lightweight backpack high voltage
electrofishing suit. U.S. Bur. Sport Fish, and Wildl. Circular 78.
8 pp.
Smith, G.F.M. and P.F. Elson. 1950. A D.C. electrical fishing apparatus.
Can. Fish Cult. 9:34-46.
Sullivan, C. 1956. Importance of size grouping in population estimates
employing electric shockers. Prog. Fish-Cult. 9:34-56.
321
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Taylor, G.N. 1957. Galvanotaxic response of fish to pulsating D.C. J.
Wildl. Management 21:201-213.
Thompson, R.B. 1959, The use of the transistorized pulsed direct current
fish shocker in assessing populations of resident fishes. In: Proc.
Thirty-ninth Ann. Conf. West. Assoc. St. Fish and Game Comm. pp. 291-
294.
Thompson, R.B. 1960. Capturing tagged red salmon with pulsed direct current.
U.S. Fish Wild!., Serv. Spec. Sci. Rept. Fish No. 355, 10 pp.
U.S. FWS. 1991. Safety Electrofishing. Jo: J.B. Reynolds. Chapter 13,
Principles and techniques of electrofishing. Fisheries Academy, U.S.
Fish and Wildlife Service, Office of Technical Fisheries Training,
Kearneysville, WV.
Vibert, R., ed. 1967. Fishing and electricity -Its applications to biology
and management. European Inland Fish. Adv. Comm. FAO, United Nations,
Fishing New (Books) Ltd. London, UK.
Vincent, R. 1971. River electrofishing and fish population estimates. Prog.
Fish-Cult. 33:163-169.
Webster, D.A., J.L. Forney, R.H.N. Gibbs, Jr., J.H. Severns, and W.F. Van
Woert. 1955. A comparison of alternating and direct electric currents
in fishery work. New York Fish Game J. 2:106-113.
Whitney, L.V. and R.L. Pierce. 1957. Factors controlling the input of
electrical energy into a fish in an electrical field, Limnol. Oceanogr.
2:55-61.
Witt, A. Jr. and R.S. Campbell. 1959. Refinements of equipment and
procedures in electrofishing. Trans. Amer. Fish. Soc. 88:33-35.
12.3 Chemical Fishing
Boccardy, J.A. and E.L. Cooper. 1963. The use of rotenone in surveying
small streams. Trans. Amer. Fish. Soc. 9:307-310.
Bone, J.N. 1970. A method for dispensing rotenone emulsions. British
Columbia fish and Wildlife Branch, Fish Management Report 62, pp. 1-3.
Dawnson, V.K., W.H. Gingerich, R.A. Davis, and P.A. Gilderhus. 1990. Rotenone
persistence in freshwater ponds: effects of temperature and sediment
adsorption. North Amer. J. Fish. Management 11:226-231.
Hester, F.E. 1959. The tolerance of eight species of warm-water fishes to
certain rotenone formulations. In: Proc. 13th Ann. Conf. Southeastern
Assoc. Game and Fish Comm.
Hocutt, C.H., P.S. Hambrick, and M.T. Masnik. 1973. Rotenone methods in a
large river system. Archives Hydrobiol. 72:245-252.
322
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Krumholz, L.A. 1950. Some practical considerations in the use of rotenone in
fisheries research. J. Wild!. Manage. 14.
Lawrence, J.M. 1956. Preliminary results on the use of potassium
permanganate to counteract the effects of rotenone on fish. Prog. Fish-
Cult. 18:15-21.
Marking, L.L. 1992. Evaluation of toxicants for the control of carp and
other nuisance fishes. Fisheries 17:6-13.
McKee, J.E. and H.W. Wolf (eds.). 1963. Water quality criteria. 2nd ed.
Calif. Water Quality Control Board Publ. 3A.
Ohio DNR. 1988. Water pollution, fish kill, and stream litter investigations
1987. Ohio Department Natural Resources, Division of Wildlife, Fountain
Square, Columbus, OH. 14 pp.
Ohio River Valley Water Sanitation Commission. 1962. Aquatic life resources
of the Ohio River, pp. 72-84.
Post, G. 1955. A simple chemical test for rotenone in water. Prog. Fish-
Cult. 17(4):190-191.
Post, G. 1958. Time vs. water temperature in rotenone dissipation. In:
Proc. 38th Ann. Conf. Game and Fish Comm. pp. 279-284.
Schnick, R.A. 1974. A review of the literature on the use of rotenone in
fisheries. La Crosse, Wis., Fish Control Laboratory, 130 pp. (Available
from NTIS, Springfield, VA 22161 as publication FWS-LR-74 15).
Schnick, R.A. 1991. Crisis in chemical and drug registration. Fisheries
16:3.
Solmon, V.E.F. 1949. History and use of fish poisons in the United States/
Dominion Wildlife Service, Ottawa, Canada.
Sowards, C.L. 1961. Safety as related to the use of chemicals and
electricity in fishery management. U.S. Fish and Wild!. Serv. Bur.
Sport Fish and Wild!., Branch Fish Management, Spearfish, SD.
Tanner, H.A. and M.L. Hayes. 1955. Evaluation of toxaphene as a fish poison.
Colo. Coop. Fish. Res. Unit, Quart, Rept. 1:31-39.
Turner, W.R. 1959. Effectiveness of various rotenone-containing preparations
in eradicating farm pond fish populations. Kentucky Dept. Fish and
Wild!. Res. Fish. Bull. No. 5, 22 pp.
Wilkins, L.P. 1955. Observations on the field use of cresol as a stream-
survey method. Prog. Fish-Cult. 17:85-86.
U.S. Dept. Interior. 1972. Recommended methods for water data acquisition.
Geol. Surv., Office Water Data Coordination.
323
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12.4 General Health, External Anomalies, Deformities, Eroded Fins, Parasites,
and Diseases
Allison, L.N., J.G, Hnath, and W,G. Yoder, 1977. Manual of common diseases,
parasites, and anomalies of Michigan fishes. Michigan Dept. Nat, Res.,
Lansing. Fish Mgmt. Rept. No. 8, 132 pp.
Amlacher, E. 1970. Textbook of fish Diseases. TFH Publication, Neptune
City, NJ.
Amos, K. 1985. Procedures for the detection & identification of certain fish
pathogens. Amer. Fish. Soc., Bethesda, MD.
Austin, B. 1988. Marine microbiology. Cambridge University Press, New York,
NY.
Austin, B. 1988. Methods in aquatic bacteriology (Modern Microbiological
Methods Ser.). John Wiley & Sons, Inc., New York, NY.
Austin, B. and D.A. Austin. 1992. Bacterial fish pathogens: Disease in
farmed and wild fish, Ellis Norwood Limited, Chichester, England.
Baumann, P.C., W.D. Smith, and W.K. Parland. 1987. Tumor frequencies and
contaminant concentrations in brown bullhead from an industrialized
river and a recreational lake. Trans. Am. Fish. Soc. 116(l):79-86.
Berra, T.M. and R.J. Au. 1978. Incidence of black spot disease in fishes in
Cedar Fork Creek, Ohio. Ohio J, Sci. 78:318-322.
Berra, T.M. and R-J. Au. 1981. Incidences of teratological fishes from Cedar
Fork Creek, Ohio. Ohio J. Sci. 81:225.
Bousfield, E.L. 1987. Amphipod parasites of fishes of Canada. Department
Fisheries and Oceans, Ottawa, Ontario, Canada, (available form
Canadian government Publishing Centre, Supply and Services Canada,
Ottawa, Ontario, Canada).
Egusa, S. 1992. Infectious Diseases. A.A. Balkema Uitgevers B.V.,
Rotterdam, Natherlands.
Ellis, A.E. 1988. Fish vaccination. Academic Press, New York, NY. 255 pp.
Esch, G.W. and T.C. Hazen. 1980. Stress and body condition in a population
of largemouth bass: implications for red-score disease. Trans, Am.
Fish. Soc. 109:532-536.
Grabda, J. 1991, Marine fish prasitology: An outline. PWB-Polish
Scientific Publishers, Warszawa, Poland (available from VCH Publishers,
New York, NY).
Herwig, N. 1979. Handbook of drugs and chemicals used in the treatment of
fish diseases. Charles C. Thomas Publisher, Springfield, IL. 272 pp.
324
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Hibiya, T. 1982. An atlas of fish histology: normal & pathological
features. Argent Laboratories, Redmond, WA.
Hoffman, G.L. 1967. Parasites of North American freshwater fishes.
Berkeley Univ. Press, Berkeley, CA.
Hoffman, G.L. and P.P. Meyer. 1974. Parasites of freshwater fishes. TFH
Publications, Neptune City, NJ.
Klemm, D.J. 1991. Taxonomy and pollution ecology of the Great Lakes Region
leeches (Annelida: Hirudinea). Michigan Academician 24:37-103.
Komanda, N. 1980. Incidence of gross malformations and vertebral anomalies
of natural and hatchery Plecoglossus altivelis, Copeia 1980:2935.
Marking, L.L. 1987. Gas supersaturation in fisheries: Causes, Concerns,,
and Cures. Fish and Wildlife Leaflet 9. Publications Unit, U.S. Fish
and Wildlife Service, Matomic Building,Room 148, Washington, DC.
Meyer, P.P. and R.A. Schnick. 1989. A review of chemicals used for the
control of fish diseases. Rev. Aquat. Sci. 2:693-710
Moller, K. and K. Anders. 1986. Diseases and parasites of marine fishes.
Verlag Moller, Kiel, Federal Republic Germany.
Margolis, L. and Z. Kubata. 1984. Guide to the parasites of fishes of
Canada. Part 1: Monogenea and Turbellaria. Can Spec. Pub!. Fish. &
Aquatic Sci. 74, Dept. Supply and Services, Canadian Government Publ.
Centre, Ottawa, Ontario, Canada,
Meyer, F,P. and G.L. Bullock. 1990. Protozoan parasites of freshwater
fishes. U.S. Fish & Wildlife Service Fish Health Bulletin 8. U.S. Fish
& Wildlife Service, Washington, DC.
Meyer, F.P. and Schnick. 1989, A review of chemicals used for the control of
fish diseases. Rev. Aquat. Sci. 1:693-710.
Meyer, F.P., J.W. Warren, and T.F. Carey. 1983. A guide to integrated fish
health management in the Great Lakes Basin. The Great Lakes Fishery
Commission, Ann Arbor, MI
Perkins, F.O. and T.C. Cheng. 1990. Pathology in marine science. Academic
Press, Inc., San diego, CA.
Pippy, J.H. and G.M. Hare. 1969. Relationship of river pollution to
bacterial infection in salmon and suckers. Trans. Am. Fish. Soc.
4:685-690.
Post, G. 1983. Textbook of fish health. TFH Publications, Inc., Neptune
City, NJ.
Reash, R.J. and T.M. Berra. 1989. Incidence of fin erosion and anomalous
325
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fishes in a polluted stream and a nearby clean stream. Water, Air, and
Soil Pollution 47:47-63.
Roberts, R.J. (ed.). 1982. Microbial diseases of fish. Academic Press, New
York, NY.
Roberts, R.J. (ed.). 1989. Fish pathology, Academic Press, Harcourt Brace
Jovanovich, San Diego, CA.
Roberts, R.J. and C.J. Shepherd. 1986. Handbook of trout and salmon
diseases. Fishing New Books, Ltd., Farnham, Surrey, England.
Rogers, W.A. and J.A. Plumb, 1977. Principal diseases of sportfish: a
fisherman's guide to fish parasites and diseases. Agric. Exp. Sta.,
Auburn Univ. Spec. Rept. Pamphlet, 17 pp.
Schaperclaus, W., H. Kulow, and K. Schreckenbach (eds.J. 1992. Fish
diseases. Volumes 1 and 2. A.A. Balkema Publishers, Rotterdam, The
Netherlands.
Sindermann. C. 1990. Principal diseases of marine fish and shellfish. Vol.
1, Academic Press, Inc., New York, NY.
Sindermann. C. 1990. Principal diseases of marine fish and shellfish. Vol.
2, Academic Press, Inc., New York, NY.
Sniezko, S.F. 1962. The control of bacterial and virus diseases of fishes.
Biological problems in water pollution, 3rd seminar. U.S. Pub!. Health
Serv. Pub. No. 999-WP-25. pp. 281-282.
Stoskopf, M.K. (ed.). 1992. Fish medicine. W.B. Saunders Co., Harcourt Brace
Jovanovich, Inc., Philadelphia, PA.
Swink, W.D. 1991. Host-size selection by parasitic sea lampreys. Trans.
Amer. Fish. Soc. 120:637-643.
Van Duijn, C. 1973. Disease of fishes. Charles C. Thomas Publisher,
Springflied, IL.
Weis, J.S. and P. Weis. 1989. Effects of environmental pollutants on early
fish development. Reviews in Aquatic Sciences 1:45-73.
Wolf, K. 1988. Fish viruses and fish viral diseases. 1988. Cornell Univ.
Press, Ithaca, New York, NY.
12.5 Fish Identification
12.5.1 General
Blair, W.F. and G.A. Moore. 1968. Vertebrates of the United States. McGraw
Hill, New York, NY.
326
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Cailliet, G.M., M.S. Love, and A.W. Ebeling. 1986. Fishes. A field manual
on their structure. Identification and Natural History. Wadsworth
Publ. Co., Belmont, CA.
Casto, J.I. 1983. Sharks of the North American waters. Texas A & M Univ.
Press, College Station, TX.
Chart. I.E. and E.P. Bergersen. 1988. Methods for long-term identification
of salmonids: a review. Publications Unit, Fish and Wildlife Service,
Patomic Building, Washington, DC.
Cummins, J.D. 1987. Index and field identification guide to the fishes of
the district of Columbia. J.D. Cummins, Government District Columbia,
Department of Consumer and Regulatory Affairs, Environmental Control
Division, Washington, DC.
Eddy, S. 1957. How to know the freshwater fishes. Wm. C. Brown Co.,
Dubuque, IA.
Eddy, S. and J.C. Underbill. 1978. How to know the freshwater fishes. Wm.
C. Brown Co., Dubuque, IA.
Eschmeyer, W.N. 1990. Catalog of the genera of recent fishes. California
Academy of Sciences, Scientific Publications Department, Golden Gate
Park, San Francisco, CA.
Gilligan, M.R. 1989. An illustrated guide to the fishes of Gray's Reef
National Marine Sanctuary. Lyons and Burford Publ., New York, NY.
Hood, d.W. and S.T. Zimmerman. 1986. The Gulf of Alaska: Physical
environment and biological resources. U.S. Government Printing Office,
Washington, DC.
Hubbs, C.L. and K.F. Lagler. 1964. Fishes of the Great Lakes region. Univ.
Mich. Press, Ann Arbor, MI.
Jordan, D.S. and B. W. Evermann. 1896-1900. The fishes of North and Middle
America; a descriptive catalogue of the species of fish-like vertebrates
found in the waters of North America, north of the Isthmus of Panama.
U.S. Natl. Mus. Bull. 47:1-331.
Jordan, D.S., B.W. Evermann, and H.W. Clark. 1930. Check list of the fishes
and fish like vertebrates of North and Middle America north of the
northern boundary of Venezuela and Colombia. U.S. Fish Wild!. Serv.,
Washington, DC.
Kendall, R.L. 1988. Taxonomic changes in North American trout names. North
Amer. J. Fisheries Management 8:389.
Kuehne, R.A. and R.W. Barbour. 1983. The American Darters. Univ. Press
Kentucky, Lexington, KY.
327
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LaMonte, F. 1958. North American game fishes. Doubleday, Garden City, New
York, NY.
Lee, D.S., C.R. Gilbert, C.H. Hocutt, R.E. Jenkins, D.E. McAllister, and R.
Stauffer. 1980. Atlas of North American freshwater fishes. Pub!.
1980-12, N. Carolina State Museum Nat. Hist., Raleigh, NC.
Lundberg, J.G. and L. A. McDade. 1990. Systematics. In: C.B. Schreck and
P.B. Moyle (eds.). Methods for fish biology. Amer. Fish. Soc.,
Bethesda, MD. pp. 65-108.
Moore, G.A. 1968. Fishes. W.F. Blair, A.T. Blair, P. Brodkorb, F.R. Kagle,
6.A. Moore (eds.). In: Vertebrates of the United States. McGraw-Hill
Book Co., New York, NY. pp. 21-165.
Morita, C.M. 1953. Freshwater fishing in Hawaii. Div. Fish Game. Dept. Land
Nat. Res., Honolulu, HI.
Nelson, J.S. 1976. Fishes of the world. John Wiley and Sons, New York, NY.
Page, L.M. 1983. Handbook of darters. TFH Pub!., Inc. Ltd., Neptune City,
NJ.
Page, L.M. and B.M. Burr. 1991. A field guide to freshwater fishes of North
America north of Mexico. The Peterson Field Guide Series, Houghton
Mifflin Co. Boston, MA.
Perlmutter, A. 1961. Guide to marine fishes. New York Univ. Press, New
York, NY.
Robins, C.R., R.M. Bailey, C.E. Bond, J.R. Brooker, E.A. Lachner, R.N. Lea,
and W.B Scott. 1990. A list of common and scientific names of fishes
from the United States and Canada. 3rd ed., Spec. Pub!. Amer. Fish.
Soc., Committee on Names of Fishes No. 12. 190 pp.
Scott, W.B. and E.J. Crossman. 1969. Checklist of Canadian freshwater fishes
with keys of identification. Misc. Publ. Life Sci. Div. Ontario Mus.
104 pp.
Smith, G.R. and R.F. Stearley 1989. The classification and scientific names
of rainbow and cutthroat trouts. Fisheries 14:4-10.
Sterba, G. 1963. Freshwater fishes of the world. Viking Press, New York,
NY.
Strauss, R.E. and C.E. Bond. 1990. Taxonomic methods: morphology. In: C.B.
Schreck and P.B, Moyle {eds.}. Methods for fish biology. Amer. Fish.
Soc., Bethesda, MD. pp. 109-140.
Thompson, J.R. and S. Springer. 1961. Sharks, skates, rays, and chimaeras.
Bur. Comm. Fish., Fish Wild!, USDI Circ. No. 119.
328
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Whitaker, J.O., Jr. 1968. Keys to the vertebrates of the eastern United
States. Burgess Publ. Co., Minneapolis, MN.
12.5.2 Larval and Immature Fishes
Auer, N.A. (ed.). 1982. Identification larval fishes of the Great Lakes
Basin with emphasis on the Lake Michigan drainage. Great Lakes Fisheries
Res. Center, Ann Arbor, MI.
Fahay, M.P. 1983. Guide to the early stages of marine fishes occurring in
the western north Atlantic Ocean, Cape Hatteras to the southern Scotian
shelf. J. Northwest Atlantic Fishery Sci. Vol. 4., Northwest Atlantic
Fisheries Organization, Bedford Institute of Oceanography, Dartmouth,
Nova Scotia.
Fritzsche, R.A. 1978. Development of fishes of the Mid-Atlantic Bight. An
atlas of egg, larval and juvenile stages. Vol. V. Chaetodontidae
through Ophidiidae, U.S. Fish and Wildlife Serv. Biol. Serv. Prog.
FWS/OBS-78/12.
Hardy, J.D., Jr. 1978. Development of fishes of the Mid-Atlantic Bight. An
atlas of egg, larval, and juvenile stages. Vol. II. Anguillidae through
Syngnathidae. U.S. Fish and Wildlife Sev. Biol. Serv. Prog. FWS/OMS-
78/12
Hardy, J.D., Jr. 1978. Development of fishes of the Mid-Atlantic Bight. An
atlas of egg, larval and juvenile stages. Vol. III. Aphredoderidae
through Rachycentridae. U.S. Fish and Wildlife Serv. Biol. Serv. Prog.
FWS/OBS-78/12.
Hoyt, R. 1988. A bibliography of the early life history of fishes. R.D.
Hoyt. Department of Biology, Western Kentucky University, Bowling Green,
KY.
Hubbs, C.L. 1943. Terminology of early stages of fishes. Copeia 4:160.
Johnson, G.D. 1978. Development of fishes of the Mid-Atlantic Bight. An
atlas of egg, larval and juvenile stages. Vol. IV. Carangidae through
Ephippidae. U.S. Fish and Wildlife Serv. Biol. Serv. Prog. FWS/OMS-
78/12.
Jones, P.M., W.D. Martin, and J.D. Hardy, Jr. 1978. Development of fishes of
the Mid-Atlantic Bight. An atlas of egg, larval and juvenile stages.
Vol. I. Acipenseridae through Ictaluridae. U.S. Fish and Wildlife
Serv. Biol. Serv. Prog. FWS/OMS-78/12.
Lippson, A.J. and R.L. Moran. 1974. Manual for identification of early
developmental stages of fishes of the Potomatic River estuary. Martin
Marietta Corp. Environ. Tech. Center, Baltimore, MD.
Mansueti, A.J. and J.D. Hardy, Jr. 1967. Development of fishes of the
329
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Chesapeake Bay region: An atlas of egg, larval and juvenile stages.
Natural Resources Inst., Univ. Maryland, College Park, MD.
Martin, F.D. and G.E. Drewry. 1978. Development of fishes of the Mid-
Atlantic Bight. An atlas of egg, larval and juvenile stages. Vol. VI.
Stramateidae through Ogcocephalidae. U.S. Fish & Wildlife Serv. Biol.
Serv. Prog. FWS/OBS-78/12.
Matarese, A.C., A.W. Kendall, D.M. Blood, and B.M. Vinter. 1989.
Laboratory guide to early life history states of northeast pacific
fishes. National Marine Fisheries Service, Seattle, WA, Northwest and
Alaska Fisheries Center. NOAA-TR-NMFS-80.
Simon, T.P. 1989. Rationale for a family-level ichthyoplankton index for use
in evaluating water quality. In: W.S.Davis and T.P. Simon (eds.).
Proceedings of the 1989 Pollution Control Biologists meeting. U.S.
Environmental Protection Agency, Chicago IL. pp. 41-65.
Snyder, D.E. 1976. Terminologies for intervals of larval fish development.
In: J. Boreman (ed.). Great Lakes fish egg and larvae identification
(proceedings of a workshop). U.S. Fish Wildlife Serv., OBS Natl. Power
Plant Team, Ann Arbor, MI. FWS/OBS-76/23. pp. 41-58.
Snyder, D.E. 1981. Contributions to a guide to the Cypriniform fish larvae
of the Upper Colorado River system in Colorado. Biol. Sci, Sedr. No. 3,
Bur. Land Management, CO.
Snyder, D.E. 1983. Fish eggs and larvae. In: L.A. Nielsen and D.L. Johnson
(eds.). Fisheries techniques. Amer. Fish. Soc., Bethesda, MD. pp.
165-198.
Wang, J.C.S. 1981. Taxonomy of the early life stages of fishes. Fishes of
the Sacramento, San Joaquin estuary and Moss Landing Harbor, Elkhorn
Slough, California. Ecological Analysts, Inc., Concord, CA.
Wallus, R., B.L. Yeager, and T.P. Simon. 1990. Reproductive biology and
early life history of fishes in the Ohio River drainage. Volume 1:
Acipenseridae through Esocidae. Tennessee Valley Authority,
Chattanooga, TN.
Wang, J.C.S. and R.J. Kernehan. 1979. Fishes of the Delaware estuaries: A
guide to the early life histories. EA Communications, Ecological
Analysts, Inc., Towson, MD.
Weinstein, M.P. (ed.). 1988. Larval fish and shellfish transport through
inlets. American Fisheries Society Symposium 3, Bethesda, MD.
12.5.3 Marine: Atlantic and Gulf of Mexico
Ackerman, B. 1951. Handbook of fishes of the Atlantic seaboard. American
Publ. Co., Washington, DC.
330
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Bearden, C.M. 1961. Common marine fishes of South Carolina. Bears Bluff
Lab. No. 34, Wadmalaw Island, SC.
Bigelow, H.B. and W.C. Schroeder. 1953. Fishes of the gulf of Maine. Fish
Bull No. 74. Fish Wild!. Serv. 53. 577 pp.
Bigelow, H.B. and W.C. Schroeder. 1954. Deep water elasmobranchs and
chimaeroids from the northwestern slope. Bull. Mus. Comp. Zool. Harvard
College, 112:37-87.
Bohlke, J.E. and C.G. Chaplin. 1968. Fishes of the Bahamas and adjacent
tropical waters. Acad, Nat. Sci. Philadelphia. Livingston Publishing
Co., Wynnewood, PA.
Bohlke, E.B., J.E. Bohlke, E. Bertelsen, W.H. Hulet, M.M. Leiby, J.E.
McCosker, J.G. Nielsen, C.H. Robins, C.R. Robins, D.G. Smith, and K.A.
Tighe, 1989. Fishes of the western North Atlantic - Part Nine
(AnguiHi formes, Saccopharynigiformes, and Leptocephali). Sears
Foundation for Marine Research, Peabody Museum of Natural History, Yale
University, New Haven, CT.
Breder, C.M., Jr. 1948. Field book of marine fishes of the Atlantic Coast
from Labrador to Texas. G.P. Putnam and Sons, New York, NY.
Casey, J.G. 1964. Angler's guide to sharks of the northeastern United
States, Maine to Chesapeake Bay. Bur. Sport Fish. Wild!. Cir. No.
179. Washington, DC.
Collette, B.B. 1988. Annotated list of the fishes of Massachusetts Bay.
U.S. Dept. Commerce, U.S. Governmnt Printing Office, Washington, DC.
Fritzssche, R.A. 1978. Development of fishes of the Mid-Atlantic Bight. An
atlas of egg, larval and juvenile stages. Vol. V. Chaetodontidae
through Ophidiidae. Biol. Serv. Prog. FWS/OMS-78/12, U.S. Fish and
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