&ER&
        United States
        Environmental Protection
        Agency
              Office of
              Water
              (WH-553)
EPA/440/4-89/001
May 1989
Rapid Bioassessment
Protocols for Use in
Streams and Rivers
                 , V
Benthic
Macroiinvertebrates and
Fish
Previously Published as EPA/444/4j89-u01


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    RAPID BIOASSESSMENT PROTOCOLS
    FOR USE IN STREAMS AND RIVERS:
BENTHIC MACROINVERTEBRATES AND FISH
                        by
                   James L.
                   Michael T. Barbour
                   Kimberly D. Porter
                    Sharon K. Gross
                   Robert M. Hughes

              (a)U.S. Environmental Protection Agency
             Assessment and Watershed Protection Division
                   401 M Street, S.W.
                  Washington, D.C. 20460

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                              Notice

This document has been reviewed in accordance  with U.S. Environmental
Protection Agency policy and approved for publication. Mention of trade names
or commercial products does not constitute endorsement or recommendation
for use.

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                                           FOREWORD
   In December 1986, U.S.  EPA's Assistant Administrator for Water initiated a major study of the Agency's sur-
face water monitoring activities. The resulting report, entitled "Surface Water Monitoring: A  Framework for
Change" (U.S. EPA 1987), emphasizes the restructuring of existing monitoring programs to better address the
Agency's current priorities, e.g., toxics, nonpoint source impacts, and documentation of "environmental results."
The study also provides specific recommendations on effecting the necessary changes. Principal among these are:

    1. To issue guidance on  cost-effective approaches to problem identification  and trend assessment.

   2. To accelerate the development and application of promising biological monitoring techniques.

   In response to these recommendations, the Assessment and  Watershed Protection Division  has developed rapid
bioassessment protocols designed to provide basic aquatic life data for planning and management purposes such as
screening, site ranking, and  trend monitoring. All of the protocols utilize fundamental assessment techniques to
generate basic information on ambient physical,  chemical, and  biological conditions.  Level of assessment and level
of effort vary with successive protocols, and  choice of a given  protocol should depend on the specific objective of
the monitoring activity and available resources. Although none of the protocols are meant to provide the  rigor  of
fully comprehensive studies, each is designed to supply pertinent,  cost-effective information when applied in the
appropriate context.
                                                      Martha G. Prothro
                                                      Director, Office of Water Regulations and Standards
                                                      U.S. EPA, Washington, D.C.
                                                      in

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                     DEDICATION TO DR. JAMES  L. PLAFKIN
       All of us in the Environmental Protection  Agency (EPA) owe  an immeasurable debt of
gratitude to Dr. James L.  Plafkin.  In addition to developing this document, Jim  was a driving
force  within EPA to increase the use  of  biology in the water pollution control program until his
untimely death on  February 6,  1990.  Throughout his decade-long career with EPA, his expertise
in ecological  assessment, his dedication, and his vision were  instrumental in  changing commonly-
held views of what  constitutes pollution and the basis for pollution control  programs.   Jim will
be remembered for his love of life,  his enthusiasm, and his  wit.  As a small token of  our  esteem,
we  dedicate this document  to his memory.
                                              IV

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                                 ACKNOWLEDGMENTS
   Dr. James L. Plafkin of the Assessment and Watershed Protection Division (AWPD) served as principal editor
and coauthor of this document. Other coauthors were consultants Michael T. Barbour, Kimberly D. Porter, and
Sharon Gross working for AWPD and Dr. Robert M. Hughes working for EPA's Corvallis Research Laboratory.

   Many others also contributed to the development of this document and deserve special thanks. First and fore-
most, the Rapid Bioassessment Workgroup. The Workgroup, composed of both State and EPA Regional biologists
(listed in Chapter 1),  was instrumental in providing a framework for the basic approach and served as primary
reviewers of various drafts. Dr. Kenneth Cummins and Dr. William Hilsenhoff provided invaluable advice on for-
mulating certain assessment metrics,  and Dr. Anthony Maciorowski and Paul Leonard supplied helpful editorial
comments on the final drafts. Special thanks go to the biologists in the field (well over a hundred) who con-
tributed their valuable time to review the document and provide constructive input. Their help in this endeavor is
sincerely appreciated.

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                                TABLE  OF  CONTENTS
                                                                                                Page

FOREWORD	     iii

ACKNOWLEDGMENTS	      v

LIST OF FIGURES	     xi

LIST OF TABLES	     xv

1. INTRODUCTION	    1-1
   1.1 Purpose of the Document	    1-1
   1.2 Development of This Document	    1-1
   1.3 A Framework for Implementing the Rapid Bioassessment Protocols  	    1-2

2. THE CONCEPT OF BIOMONITORING	    2-1
   2.1 Biosurveys, Bioassays, and Chemical Monitoring	    2-1
   2.2 Use of Different Taxonomic Groups in Biosurveys	    2-2
   2.3 Station Siting	    2-3
   2.4 Importance of Habitat Assessment	    2-4
   2.5 The Ecoregion Concept	    2-4
   2.6 Data Management and Analysis	    2-6
       2.6.1 Integration into BIOS	    2-6
       2.6.2 Computerizing Field Data for Calculation of the Metrics	    2-6
   2.7 Benthic Community Considerations	    2-6
       2.7.1 Seasonality for Benthic Collections	    2-6
       2.7.2 Benthic Sampling Methodology	    2-9
            2.7.2.1  Natural and Artificial  Substrates 	    2-9
            2.7.2.2  Single and Multiple Habitat Sampling	   2-11
            2.7.2.3  Sampling Coarse Paniculate Organic Material (CPOM) 	   2-11
       2.7.3 Benthic Sample Processing and Enumeration	   2-12
       2.7.4 Benthic Environmental  Tolerance Characterizations	   2-12
   2.8 Fish Community Considerations	   2-12
       2.8.1 Seasonality for Fish Collections	   2-12
       2.8.2 Fish Sampling Methodology	   2-13
            2.8.2.1  Use of Electrofishing,  Seining, and Rotenoning	   2-13
            2.8.2.2  Sampling Representative Habitat	   2-13
       2.8.3 Fish Sample Processing and Enumeration	   2-14
       2.8.4 Fish Environmental Tolerance Characterizations 	   2-14

3. OVERVIEW OF PROTOCOLS  AND SUMMARY OF COMPONENTS	    3-1
   3.1 Summary of the Protocols	    3-1
   3.2 Objectives of the Protocols	    3-1
   3.3 Level of Effort  and Investigator Expertise	    3-1

4. QUALITY ASSURANCE/QUALITY CONTROL 	    4-1
   4.1 Program Description	    4-1
   4.2 Data Quality Objectives	    4-1
   4.3 Quality Assurance Program Plans  and Project Plans	    4-1
                                                 vn

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   4.4 EPA Responsibilities	    4-2
   4.5 Importance of QA/QC for Rapid Bioassessments	    4-2

5. HABITAT ASSESSMENT AND PHYSICOCHEMICAL PARAMETERS	    5-1
   5.1 Physical Characteristics and Water Quality	    5-1
       5.1.1 Physical Characterization	    5-1
       5.1.2 Water Quality	    5-3
   5.2 Habitat Assessment	    5-3
       5.2.1 Primary Parameters—Substrate and Instream Cover  	    5-4
       5.2.2 Secondary Parameters—Channel Morphology 	    5-7
       5.2.3 Tertiary Parameters—Riparian and Bank Structure 	    5-7

6. BENTHIC MACROINVERTEBRATE BIOSURVEY AND DATA ANALYSIS	    6-1
   6.1 Rapid Bioassessment Protocol I—Benthic Macroinvertebrates	    6-1
       6.1.1 Field Methods	    6-1
       6.1.2 Data Analysis Techniques	    6-1
   6.2 Rapid Bioassessment Protocol II—Benthic Macroinvertebrates	    6-4
       6.2.1 Field Methods	    6-4
             6.2.1,1  Sample Collection	    6-4
             6.2.1.2  Sample Sorting and Identification	    6-7
       6.2.2 Data Analysis Techniques	   6-10
   6.3 Rapid Bioassessment Protocol HI—Benthic  Macroinvertebrates  	   6-16
       6.3.1 Field Methods	   6-16
             6.3.1.1  Sample Collection	   6-16
             6.3.1.2  Field Processing of the CPOM Sample	   6-18
       6.3.2 Lab Methods	   6-18
             6.3.2.1  Sample Sorting and Identification	   6-18
       6.3.3 Data Analysis Techniques	   6-19
   6.4 Results of a Pilot Study Conducted on the Ararat  and Mitchell Rivers, North Carolina 	   6-26
       6.4.1 Introduction	   6-26
       6.4.2 Methods	   6-28
             6.4.2.1  Field Collections	   6-28
             6.4.2.2  Laboratory Processing	   6-30
             6.4.2.3  Quality Assurance	   6-30
       6.4.3 Bioclassification of Stations Based on the North Carolina DEM Protocol	   6-30
       6.4.4 Selection of Metrics	   6-32
       6.4.5 Comparison of  Multihabitat vs. Single Habitat Collections	   6-33
       6.4.6 Evaluation of the 100-Organism Subsample	   6-38
       6.4.7 Family-Level vs. Species-Level Identification	   6-38
       6.4.8 Integrated Bioassessment  	   6-39

7. FISH BIOSURVEY AND DATA ANALYSIS  	    7-1
   7.1 Rapid Bioassessment Protocol IV—Fish  	'.	    7-1
       7.1.1 Design of Fish  Assemblage Questionnaire Survey	    7-1
       7.1.2 Response Analysis	    7-1
   7.2 Rapid Bioassessment Protocol V—Fish	    7-5
       7.2.1 Field Survey Methods	    7-5
             7.2.1.1  Sample Collection	    7-5
             7.2.1.2  Sample Processing	    7-9
       7.2.2 Data Analysis Techniques	    7-9
             7.2.2.1  Description  of IBI Metrics	   7-12
   7.3 Results of Pilot Studies in Ohio and Oregon	   7-20
       7.3.1 Methods	   7-20
       7.3.2 Results and  Interpretation	'.	   7-20
                                                   Vlll

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8.  INTEGRATION OF HABITAT, WATER QUALITY, AND BIOSURVEY DATA	    8-1
   8.1 The Relationship Between Habitat Quality and Biological Condition	    8-1
   8.2 Bioassessment Technique	    8-3
   8.3 An Integrated Assessment Approach	    8-5
   8.4 Case Study	   8-16

REFERENCES                                                                                 R-l

APPENDIX A: GUIDANCE FOR USE OF FIELD AND LABORATORY DATA SHEETS
              A. 1 Guidance  for Header Information	    A-l
              A.2 Guidance  for Biosurvey Field Data Sheet for Behthic RBPs I, II, and III	    A-l
              A.3 Guidance  for Impairment Assessment Sheet for RBPs I, II, III, and V	    A-2
              A.4 Guidance  for Data Summary Sheet for Benthic RBPs II and III	    A-2
              A.5 Guidance  for Laboratory Bench Sheet for Benthic RBP III	    A-4
              A.6 Guidance  for Field Collection Data  Sheet for Fish RBP V	    A-4
              A.7 Guidance  for Data Summary Sheet for Fish RBP V	    A-5

APPENDIX B: RAPID BIOASSESSMENT SUBSAMPLING METHODS FOR BENTHIC PROTOCOLS
              II AND III (100-Organism Count Technique)
              B.I Rapid Bioassessment Subsampling Methods for Protocol II	    B-l
              B.2 Rapid Bioassessment Subsampling Methods for Protocol III	    B-l

APPENDIX C: FAMILY  AND  SPECIES-LEVEL  MACROINVERTEBRATE  TOLERANCE
              CLASSIFICATIONS
              C. 1 Family-Level Tolerance Classification	    C-l
              C.2 Genus/Species-Level Tolerance  Classification .•	    C-l
              C.3 References for Determining Family and Species-Level Tolerance Classifications	    C-l
              C.4 A Partial Listing of Agencies That Have Developed Tolerance Classifications and/or Biotic
                  Indices	    C-4

APPENDIX D: TOLERANCE, TROPHIC GUILDS, AND ORIGINS OF SELECTED FISH SPECIES
              D.I Species-Level Fish Tolerance, Trophic, and Origin Classifications	    D-l
              D.2 Selected References for Determining Fish Tolerance, Trophic, Reproductive, and Origin
                  Classifications	    D-7
              D.3 Agencies  Currently Using or Evaluating Use  of the IBI for Water Quality Investiga-
                  tions 	   D-l 1
                                               IX

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                                      LIST  OF FIGURES
                                                                                                       Page

1.3-1  Bioassessment decision matrix	      1-4

2.5-1  Ecoregions of the conterminous United States	      2-5

2.5-2  Flowchart illustrating potential delineation of reference sites within an ecoregion	      2-7

2.6-1  Header information used for documentation and identification of sampling stations	      2-8

2.7-1  Classification of U.S. climatological regions	     2-10

3.2-1  Overview of the five bioassessment approaches and their primary objectives  	      3-4

5.1-1  Physical  Characterization/Water  Quality  Field Data  Sheet for  use with all Rapid  Bioassessment
       Protocols	      5-2

5.2-1  Habitat Assessment Field Data Sheet for  use with all  Rapid Bioassessment Protocols  	      5-5

6.1-1  Biosurvey Field Data Sheet for use with  Rapid Bioassessment Protocol I 	      6-2

6.1-2  Impairment Assessment Sheet for use with macroinvertebrate Rapid Bioassessment Protocols	      6-3

6.2-1  Biosurvey Field Data Sheet for use with  Rapid Bioassessment Protocol II	      6-6

6.2-2  Data Summary Sheet suggested  for use  in  recording benthic data  utilized in Rapid  Bioassessment
       Protocol  II	      6-8

6.2-3  Flowchart of bioassessment approach advocated for Rapid Bioassessment Protocol II	     6-12

6.3-1  Biosurvey Field Data Sheet for use with  Rapid Bioassessment Protocol III	     6-17

6.3-2  Laboratory Bench Sheet suggested for use in recording benthic data utilized in Rapid  Bioassessment
       Protocol  III	     6-20

6.3-3  Data Summary Sheet suggested  for use  in  recording benthic data  utilized in Rapid  Bioassessment
       Protocol  III	     6-23

6.3-4  Flowchart of bioassessment approach advocated for Rapid Bioassessment Protocol III	     6-27

6.4-1  Pilot study station locations, Ararat River,  North Carolina, September 1986	     6-29

6.4-2  Cluster analysis results for benthic community metrics, based on 100-organism subsamples from riffle
       samples collected on the Ararat and Mitchell Rivers	     6-33

6.4-3  Comparison of taxa richness  for all  field-sorted  samples collected  on the Ararat  and Mitchell
       Rivers 	     6-36

6.4-4  Station cluster analysis  results for field-sorted riffle  samples collected on the Ararat and Mitchell
       Rivers 	     6-37

                                                     xi

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6.4-5  Station cluster analysis results for 100-organism subsamples from riffle samples collected on the Ararat
       and Mitchell  Rivers  	     6-37

6.4-6  Station cluster analysis results for 300-organism subsamples from riffle samples collected on the Ararat
       and Mitchell  Rivers  	     6-38

6.4-7  Cluster  analysis results  for benthic community  metrics,  based on  family-level identifications  of
       100-organism subsamples from riffle samples collected on the Ararat and Mitchell Rivers	     6-39

6.4-8  Station cluster analysis results for benthic community metrics, based on family-level identifications of
       100-organism subsamples from riffle samples collected on the Ararat and Mitchell Rivers	     6-39

7.1-1  Fish assemblage questionnaire for use with Rapid Bioassessment Protocol IV	      7-2

7.2-1  Impairment Assessment Sheet for use  with fish Rapid Bioassessment Protocol V	      7-7

7.2-2  Flowchart of  bioassessment approach advocated for Rapid Bioassessment Protocol V	      7-8

7.2-3  Fish Field Collection Data Sheet for use with  Rapid Bioassessment Protocol V	     7-10

7.2-4  Total number of fish species versus  watershed  area for Ohio regional reference sites	     7-16

7.2-5  Data Summary  Sheet for Rapid  Bioassessment Protocol V  	     7-19

7.3-1  Locations of  regional reference sites in Ohio	     7-20

7.3-2  Locations of  sampling sites on the mainstem Willamette River, Oregon	     7-21

7.3-3  Index  of Biotic  Integrity  scores by Ohio ecoregion	     7-22

7.3-4  Dominant Ohio fish  species by ecoregion  	     7-23

7.3-5  Patterns  in mainstem Willamette  River  fish assemblages  as  revealed by  detrended correspondence
       analysis  	/:.	     7-25

7.3-6  Longitudinal  trends in Index of Biotic Integrity and nitrate in the Willamette River	     7-26

8.1-1  The relationship between habitat and biological condition	      8-1

8.1-2  Relationship of habitat quality and biological condition in the context of water quality  	      8-2

8.2-1  Range of sensitivities of Rapid Bioassessment Protocol II and III benthic metrics in assessing biological
       condition in response to  organics and  toxicants	      8-3

8.2-2  Range of sensitivities of  Rapid  Bioassessment  Protocol  V  fish  metrics in assessing biological
       condition	      8-4

8.3-1  Evaluation of habitat at a site-specific control relative to that at a regional reference	      8-5

8.3-2  Evaluation of water quality effects	      8-7

8.3-3  Evaluation of biological  impairment due to reversible habitat  alterations	      8-8

8.3-4  Evaluation of an alternative site-specific control station	     8-10
                                                        XII

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8.3-5  Bioassessment using a site-specific control station  	      8-11

8.3-6  Bioassessment using a regional reference	      8-15

8.4-1  The relationship between habitat quality and benthic community condition at the North Carolina pilot
       study site	      8-19

8.4-2  Pilot study  results applied  to the theoretical habitat versus biological condition curve	      8-19
                                                        Xlll

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                                       LIST  OF  TABLES
                                                                                                      Page

3.1-1  Comparison of Rapid Bioassessment Protocols	     3-2

6.2-1  Criteria for characterization of biological condition for Rapid Bioassessment Protocol II	    6-11

6.3-1  Criteria for characterization of biological condition for Rapid Bioassessment Protocol III	    6-21

6.4-1  Bioclassification results for North Carolina DEM multihabitat benthic samples collected from the Ararat
       and Mitchell Rivers, 23-24 September 1986	    6-31

6.4-2  Taxa richness,  by group, for samples collected by North Carolina DEM from the Ararat and Mitchell
       Rivers 	    6-32

6.4-3  Metric values,  percent comparison, and bioassessment scores for benthic pilot study results: 100-, 200-,
       and 300-organism subsample  data	    6-34

6.4-4  Metric values,  percent comparison, and bioassessment scores for benthic pilot study results: EA field-
       sorted and family level identification data  .. .	    6-35

6.4-5  Summary of the bioclassification derived from an analysis of samples collected from the Ararat and
       Mitchell  Rivers	    6-40

7.2-1  Regional variations  of IBI metrics	    7-13

7.3-1  Collection data for two Ohio  ecoregion reference sites	    7-21

7.3-2  Scoring criteria and IBI and IWB scores for two Ohio ecoregion reference sites 	    7-22

7.3-3  Collection data (number of individuals) for two sites on the  Willamette River, Oregon	    7-24

7.3-4  Scoring criteria and IBI and IWB scores for two sites on the Willamette River, Oregon	    7-25

8.3-1  Bioassessment conclusions  relative to use of a site-specific control or regional reference	    8-12

8.4-1  Summary of  habitat  assessment  scoring  for  Ararat  and  Mitchell Rivers  benthic  case-study
       data 	    8-17

8.4-2 -Summary of metric values, percent comparison, and bioassessment scores for Ararat and Mitchell Rivers
       benthic case-study  data	    8-18

C-l    Tolerance values for families  of stream arthropods in the western Great Lakes region	    C-2

C-2    Tolerance values for some macroinvertebrates not included in Hilsenhoff (1982, 1987b)	    C-3

D-l    Tolerance, trophic guilds, and origins of selected fish species	     D-l
                                                     xv

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                                     1.  INTRODUCTION
         1.1  PURPOSE OF THE
                DOCUMENT
   The primary purpose of this document is to pro-
vide States with a practical technical reference for
conducting cost-effective biological assessments of
lotic systems. The protocols presented are not  neces-
sarily intended to replace those already  in use by State
agencies. Instead, they provide options for agencies
that wish to implement rapid biological  assessment
techniques. Three macroinvertebrate and two fish pro-
tocols are presented:  Benthic Rapid Bioassessment
Protocol I (RBP  I) and fish Rapid Bioassessment Pro-
tocol IV (RBP IV) are cost-effective screening proce-
dures that provide some supporting data; benthic
Rapid Bioassessment Protocol II (RBP II) can  help set
priorities for more intensive evaluations; and benthic
Rapid Bioassessment Protocol III (RBP  III) and  fish
Rapid Bioassessment Protocol V (RBP V) are  progres-
sively more rigorous and provide more  confirmational
data, but also require more resources. The choice of a
particular  protocol should depend  on the purpose of
the bioassessment, the need to document conclusions
with confirmational data, the degree of  discrimination
desired, and available resources. Although the benthic
protocols were designed and tested in wadable fresh-
water streams rather  than large rivers (or lakes,  estu-
aries, or marine  systems), the fundamental approach
should be applicable  to large freshwater rivers as well.
The fish protocols were validated  in freshwater
streams and large rivers and are applicable to  both.
   The original rapid bioassessment protocols  were
designed as inexpensive screening tools  for determin-
ing if a stream is supporting or not supporting a
designated aquatic life use. The basic information
generated  would  enhance the coverage of broad
geographical assessments,  such as State  and National
305(b) Water Quality Inventories.  However, members
of a  1986 benthic Rapid Bioassessment Workgroup
and reviewers of this document indicated that the
rapid bioassessment protocols  can  also be applied to
otrler program  areas, for example:

• Characterizing the  existence and severity of  use
  impairment
• Helping to identify sources  and  causes of use
  impairment
• Evaluating the effectiveness of control actions
• Supporting use attainability studies
• Characterizing regional biotic components

Therefore, the scope of this guidance might now be
considered applicable to a wider range of planning
and management purposes than originally envisioned,
i.e., they may be appropriate for priority setting,  point
and nonpoint-source evaluations,  use attainability  anal-
yses, and trend monitoring, as well as initial
screening.
        1.2  DEVELOPMENT OF
           THIS DOCUMENT
   This document was developed in two phases. The
first phase centered  on the development and refine-
ment of the benthic  rapid bioassessment protocols.
The second phase involved the addition of analogous
protocols pertinent to  the assessment of fish
communities.
   The benthic protocols were developed by con-
solidating procedures in  use by various State water
quality agencies. In  1985, a survey  was conducted to
identify States that routinely perform screening-level
bioassessments and believe that such efforts are
important to their monitoring  programs. Guidance
documents and field methods  in common use were
evaluated in  an effort to  identify successful bioassess-
ment methods that use different levels  of effort.  Origi-
nal survey materials and information obtained  from
direct personal contacts were  used to develop the draft
protocols.
   Missouri Department of Natural  Resources and
Michigan Department of Natural Resources both use
the "stream  walk" approach upon which the screening
protocol (RBP I) in  this  document is based. The sec-
ond protocol  (RBP II) is more time and labor inten-
sive, incorporating field  sampling and  family-level
taxonomy, and is a less intense version of RBP HI.
The concept of family-level  taxonomy  is based on  the
approach used by the  Virginia State Water Control
Board. The third protocol (RBP III) incorporates
certain aspects of the  methods used  by the North
Carolina Division of Environmental  Management and
the New York Department of  Environmental Conser-
vation and is the most rigorous of the  three
approaches.

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   A workgroup of State and U.S. EPA Regional biol-
ogists (listed below) was formed to review and refine
the draft benthic protocols. The Rapid Bioassessment
Workgroup included biologists using the State methods
described above and biologists from other regions
where pollution sources and aquatic systems differed
from those areas for which the draft protocols were
initially developed.

EPA

James Plafkin,  AWPD
Michael Bilger, Region  I
Michael Bastian, Region VI
William Wuerthele, Region VIII
Evan Hornig, Region  X

STATES

Brenda Sayles,  Michigan DNR
John Rowland,  Missouri DNR
Robert Bode, New York DEC
David Lenat, North Carolina DEM
Michael Shelor, Virginia SWCB
Joseph Ball, Wisconsin  DNR

   The rapid bioassessment protocols  for benthos
presented here include modifications discussed in the
workgroup's first meeting held in 1986, as well as
comments on a subsequent draft.  This document also
includes results of a field validation study (Section 6.4)
conducted with the North Carolina Department of
Environmental Management to examine certain
methodological  issues highlighted in the workgroup's
review (Chapter 2). In addition,  these protocols and
the concept of "rapid" bioassessment have been dis-
cussed at the 1986, 1987, and 1988 annual meetings of
the North American Benthological Society and in per-
sonal communications with Dr. Kenneth Cummins,
Dr. William Hilsenhoff, Dr. James Karr, and Dr.
Vincent Resh.
   In response  to a number of comments received
from State and  EPA personnel  on an earlier version  of
the rapid bioassessment protocols, a set of fish pro-
tocols was also developed. Fish protocol V is based
on Karr's work (1981) with the Index of Biological
Integrity (IBI), Gammon's Index of Well Being (1980),
and standard fish population assessment models, cou-
pled with certain modifications for implementation in
different geographical regions.  Ohio EPA has devel-
oped biological criteria using the IBI and Index of
Well Being (IWB), and a substantial database on their
use for site-specific fish assessments exists.
       1.3  A FRAMEWORK FOR
    IMPLEMENTING  THE RAPID
   BIOASSESSMENT PROTOCOLS
   The rapid bioassessment protocols advocate an
integrated assessment, comparing habitat (e.g., physi-
cal structure, flow regime) and biological measures
with empirically defined reference conditions (Figure
1.3-1). Reference conditions are established through
systematic monitoring of actual sites that represent the
natural range of variation in "least disturbed" water
chemistry, habitat, and biological condition. Of these
three components of ecological integrity, ambient
water quality may be the most difficult to characterize
because of the complex array  of chemical constituents
(natural and otherwise) that affect it. Therefore, the
implementation framework presented below first
describes the development  of an empirical relationship
between habitat quality and biological condition, then
refines this relationship for a given region. As addi-
tional information is obtained from systematic
monitoring of potentially impacted and site-specific
control sites, the predictive power of the empirical
relationship  is enhanced. Once the relationship
between habitat and biological potential is understood,
water quality impacts can be objectively discriminated
from habitat effects and control efforts can be focused
on the most important source of impairment.
                                                    1-2

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Explanatory Notes—
Bioassessment Implementation Framework
   The following notes describe the implementation
framework of the RBPs presented in Figure 1.3-1;
each note corresponds to a similarly numbered  ele-
ment  in the flow diagram.  The reader should examine
the figure first,  then refer back to the numbered notes
for explanations.
1. The "reference site" (RS) should represent a data-
   base consisting of the best attainable physical
   habitat, water chemistry, and biological parameters
   for specific environmental conditions. Acceptable
   ranges for the habitat and biological  parameters of
   concern are based on this reference database.
       In the RBP assessment scheme, selected
   parameters are integrated to define generic habitat
   categories and bioclassifications. The integrated
   characterizations describe important attributes of
   the designated use and  represent criteria for
   attainment/non-attainment of the designated  use.
   Figure 1.3-1  also illustrates how designated  uses
   and criteria may be established or  refined as
   ambient  monitoring activities  proceed, and how
   new data are  incorporated into the reference
   database.
       Considerable effort may be required  initially to
   identify reference  sites and the habitat and biologi-
   cal characteristics of a specified aquatic life use.
   Alternatively, data required to define new  or
   refined use characterizations and assessment
   criteria could be collected through implementation
   of an effective ambient monitoring program. How-
   ever, when the initial reference database includes a
   spectrum of "least disturbed"  habitats and con-
   comitant  biotic conditions, the need  for site-specific
   controls may be greatly reduced. The value of a
   comprehensive reference database becomes  more
   evident with progression through the implementa-
   tion framework.
 2. The purpose of the habitat assessment is to  deter-
    mine whether "IS" (impaired site) has the potential
   to support a biological  community comparable to
    that of the reference (see note 6).
 3. Generally applicable ranges for several important
    habitat characteristics are incorporated into  the
    habitat assessment field sheets (Figure 5.2-1) and
    the  habitat evaluation can be  made quickly  onsite.
    However, preliminary reconnaissance is especially
    helpful when impaired  site habitat (HIS) proves to
    be much lower in quality than reference habitat
    (HRS) and an evaluation of reversible habitat alter-
    ations ("attainability")  may also be necessary.
    Reconnaissance information allows planning for the
    additional work needed to characterize more
    appropriate reference sites.
 4. In the early stages of developing assessment
    criteria for a given  aquatic life use,  HIS may often
    appear degraded relative to the HRS database. The
    likelihood of such an outcome  is proportional to
    the  richness of the initial HRS database. As more
   potentially  impacted stations are assessed,  however,
   certain stations will be shown to support:
   —Biological communities equivalent to the refer-
     ence sites despite apparent habitat deficiencies.
     Information from such sites will enrich the refer-
     ence database and broaden the applicability of
     the use designation.
   —A  relatively degraded community that is  limited
     by intrinsic or irreversible habitat constraints. In
     this case, the original use is not attainable, and
     data collected from such a site should be used to
     revise  the use designation.
5.  The  robustness of the comparison between the bio-
   logical condition at the impaired site  (BIS) and that
   at the reference (BRS) is  limited by the rigor
   of the assessment  procedure used (e.g.,  many
   versus few replicates) and the scope of the overall
   assessment (i.e., the number of biological  commu-
   nity  segments actually evaluated). The comparison
   of BIS and BRS is useful for  detecting or  confirm-
   ing appreciable impact to the biotic community
   and  may be insensitive to certain subtle and/or
   threshold effects.
6.  If BIS = BRS, there is no detectable impairment.
   This conclusion assumes no overriding limitations
   on the biological potential of "IS" relative to  "RS
   that  are  not accounted for by the previous habitat
   comparison (see Note 2). Factors that could
   uniquely affect "IS"  are discussed in Section 2.3.
   For  example, stations  "RS" and  "IS" may be
   located on a first  order stream with primary
   organic inputs from a coniferous forest. In this sit-
   uation, certain characteristics of the benthic com-
   munity, such as taxa richness, may actually
   increase with organic  enrichment from point source
   discharges rather than decrease as otherwise
   expected. This atypical situation should be assessed
   as if HIS-(-(Reversible Habitat Alterations)< HRS
   (Step 7).
7. The HIS + (Reversible Habitat Alterations) versus
   HRS comparison  amounts to  a simplistic use
   attainability analysis (UAA) that only considers
   habitat. The comparison involves scaling up the
   observed habitat parameter values to the extent that
   they might be feasibly improved.  For example,
   bank stability, bank vegetation, and streamside
   cover could be greatly enhanced by fencing a pas-
   ture and planting  trees, whereas other parameters
   may be unalterable. This "mini-UAA" can help to
   assess site-specific potential  in the determination of
   actual impairment. If HIS and HRS  are potentially
   equivalent, then use impairment can  be appropri-
   ately assessed in terms of the resident biota. If HIS
   and  HRS are not  equivalent even when reversible
   habitat alterations are considered, biological effects
   may not be independent of habitat constraints.
   These potential scenarios are discussed in  more
   detail  in Chapter  8, Integration of Habitat, Water
   Quality, and Biosurvey Data. The approach to con-
   ducting a habitat assessment and bioassessment  is
   discussed  in Chapters 5, 6, and 7.
                                                     1-3

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                                     Assessment of
                           Impairment of Biological Integrity
                    (Not Human Health, Recreation or Aesthetics)*1)
               Potentially Impacted/
                 Impaired Site (IS)
            HIS = Habitat; BIS = Biological
                   Classification
   vs
     Reference Site (RS)
 for a given Designated Use;
HRS = Habitat; BRS = Biological
   Classification (Criterion)
  HIS = HRS
                                      HIS < HRS
                                         (4)
                                         BIS < BRS
  BIS = BRS
     (5)
 Bioimpact
Impairment
   at IS
    (6)
                                                                                HIS+
                                                                            (Revs Hab Alts)
                                                                               vsHRS
                                                                                 (7)
 No Bioimpact
No Impairment
    at IS
                                         See
                                       Chapter
                                          8
                Figure 1.3-1. Bioassessment decision matrix. (Numbers in
                             parentheses refer to points of discussion in text).
                                           1-4

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                   2.  THE  CONCEPT OF  BIOMONITORING
   2.1  BIOSURVEYS,  BIOASSAYS,
  AND CHEMICAL MONITORING
   The water quality-based approach to pollution
assessment requires various types of data. Biosurvey
techniques, such as the rapid bioassessment.protocols,
are best used for detecting aquatic life impairments
and assessing their relative severity. Once an  impair-
ment  is detected, however, additional chemical and
biological (toxicity) testing is usually necessary to
identify the causative agent and its source and to
implement appropriate mitigation (U.S. EPA 1985).
Following mitigation, biosurveys are important for
evaluating the effectiveness of such control measures.
   Biosurveys may be used within a planning and
management  framework to prioritize water quality
problems for more stringent assessments and to docu-
ment  "environmental  recovery" following control
action.  Some of the advantages of using biosurveys  for
this type of monitoring are:

1. Biological communities reflect overall ecological
   integrity (i.e., chemical, physical, and biological
   integrity)., Therefore, biosurvey results directly
   assess the status of a waterbody relative to the
   primary goal of the Clean Water Act.
2. Biological communities integrate the effects of
   different pollutant  stressors and thus provide a
   holistic measure of their aggregate  impact. Com-
   munities also integrate  the stresses  over time  and
   provide an ecological measure of fluctuating
   environmental conditions.  Assessing the integrated
   response of biological communities to highly
   variable pollutant inputs offers a particularly useful
   approach  for monitoring nonpoint-source impacts
   and the effectiveness of certain Best Management
   Practices.
3. Routine monitoring of biological communities  can
   be relatively inexpensive, particularly when com-
   pared to the cost of assessing toxic pollutants,
   either chemically  or with toxicity tests (Ohio  EPA
   1987a).
4. The  status of biological communities is of direct
   interest to the public as a measure  of a pollution
   free  environment, while reductions in chemical
   pollutant loadings are not as readily understood by
   the layman as positive environmental results.
5.  Where criteria for specific ambient impacts do not
   exist (e.g., nonpoint-source impacts that degrade
   habitat), biological communities may be the only
   practical means of evaluation.

   Biosurvey methods have a long-standing history of
use for "before and after" monitoring. However, the
intermediate steps in pollution control, identifying
causes and limiting sources, require information of a
different  type—chemical, physical, and/or additional
biological data. These data are needed to:

1.  Identify the specific stress agents causing impact.
   This may  be a relatively simple task; but, given
   the array of potentially important pollutants (and
   their possible combinations),  it is likely to be both
   difficult and costly. In situations where specific
   chemical stress agents are either poorly understood
   or too varied to  assess individually, toxicity tests
   can be used to focus specific chemical investiga-
   tions or to characterize generic stress agents (e.g.,
   whole effluent toxicity).
2.  Identify and limit the specific sources of these
   agents. Although biosurveys can be used to help
   locate the likely origins of impact, chemical anal-
   yses and/or toxicity tests are usually necessary to
   confirm the responsible sources and develop
   appropriate discharge limits.
3.  Design appropriate treatment to meet the
   prescribed limits and monitor compliance. Treat-
   ment  facilities are designed to remove  identified
   chemical constituents with a specific efficiency.
   Chemical  data are therefore required to construct
   such facilities and evaluate treatment effectiveness.
   To some degree, a biological endpoint resulting
   from toxicity testing can also be.used to evaluate
   the effectiveness of prototype treatment schemes
   and can serve  as a design pa-ameter. In most
   cases,  these same  parameters are limited in dis-
   charge permits and,  after  controls  are  in place, are
   used to monitor for compliance. Where discharges
   are not controlled  through a permit system (e.g.,
   nonpoint-source  runoff, combined  sewer outfalls,
   and dams) compliance must be assessed in terms
   of ambient standards.
   Effective implementation of the water quality-based
approach requires that various monitoring techniques
be considered within a larger context  of water
resource  management. Both biological and chemical
                                                    2-1

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methods play critical roles in a successful pollution
control  program. They should be considered com-
plementary rather than mutually exclusive approaches
that will enhance overall program effectiveness when
used appropriately.
       2.2  USE  OF DIFFERENT
         TAXONOMIC GROUPS
             IN BIOSURVEYS
   The bioassessment techniques presented in this
document focus on the evaluation of water quality,
habitat, and benthic macroinvertebrate and fish com-
munity parameters. Many State water quality agencies
employ trained and experienced benthic biologists,
have accumulated considerable background data on
macroinvertebrates, and consider benthic surveys a
useful  assessment tool. However, water quality stan-
dards,  legislative mandate, and public opinion are
more directly related to the status of a waterbody as a
fishery resource. For this reason, separate protocols
were developed for fish and were incorporated as
Chapter 7 in this document. The fish survey protocol
is based largely on James Karr's IBI (Karr 1981; Karr
et al. 1986; Miller et al.  1988a), 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 com-
mon element to the fish  and benthic rapid bioassess-
ment approaches.
   Although no methods are presented here for con-
ducting algal assessments, algal communities are also
useful  for water quality monitoring. They represent
another trophic level, exhibit a different range of sen-
sitivities, and will often  indicate effects only indirectly
observed in the benthic and fish communities. As  in
the benthic macroinvertebrate  and fish communities,
integration of structural/compositional and functional
characteristics provides the best means of assessing
impairment (Rodgers et  al.  1979).
   Algal community structural/compositional  analyses
may be taxonomic  or non-taxonomic. Taxonomic anal-
yses (e.g., diversity indices, taxa richness, indicator
species) are commonly used,  and are described in
studies by  Rodgers et al. (1979), Weitzel (1979),
Palmer (1977), and Patrick (1973). Non-taxonomic
measures, such as  biomass and chlorophyll, can also
be useful for detecting effects not indicated  by taxo-
nomic analysis. For example,  toxic pollutants may
cause  sublethal (i.e., reproductive) effects which
would not immediately be detected by taxonomic ana-
lyses such  as  taxa richness, but would be indicated by
low biomass (Patrick 1973). A summary of non-
taxonomic measurements is presented in Weitzel
(1979).
   Functional  aspects of algal communities, such as
primary productivity rates, can also be assessed.
These analyses, which  are described in Rodgers et  al.
1979,  can be performed at the taxonomic  level  (e.g.,
determination  of species colonization rate) or at the
non-taxonomic level (e.g., community respiration) to
evaluate effects of toxicants or nutrient  enrichment.
   In determining the taxonomic group or groups
appropriate for a  particular biomonitoring situation,
the advantages of using each  taxonomic group  must be
considered along  with the objectives of the program.
Some of the advantages of using macroinvertebrates,
fish, and algae in a biomonitoring program are
presented  in this section. References for this list are
Cairns and Dickson 1971;  Karr 1981; U.S. EPA 1983;
Hughes et al.  1982; American Public Health Associa-
tion et al. 1971; Patrick 1973; Rodgers et  al. 1979; and
Weitzel 1979.

Advantages of Using Benthic Macroinvertebrates
1.  Macroinvertebrate communities are good indicators
   of localized conditions.
   • Because  many benthic macroinvertebrates have
     limited migration patterns or a sessile mode of
     life,  they are particularly well suited for assess-
     ing site-specific impacts (upstream-downstream
     studies).
2. Macroinvertebrate communities integrate the effects
   of short-term environmental variations.
   •  Most species have a complex life cycle of
     approximately 1 year or more. Sensitive  life
      stages will  respond quickly to stress;  the overall
      community  will respond more slowly.
3. Degraded conditions can often be detected by an
   experienced biologist with only a cursory examina-
   tion of the  macroinvertebrate community.
   •  Macroinvertebrates are relatively easy to  identify
      to family; many "intolerant" taxa can be identi-
      fied to lower taxonomic levels with ease.
4.  Sampling is relatively easy,  requires few people
    and inexpensive gear, and has no detrimental effect
    on the resident biota.
5.  Benthic macroinvertebrates serve as a primary food
    source for  many recreationally and commercially
    important fish.
6.  Benthic macroinvertebrates are abundant in most
    streams.
    •  Many small streams (1st and 2nd order), which
      naturally support a diverse  macroinvertebrate
      fauna, only support a limited fish fauna.
7.  Most State water quality agencies that routinely
                                                      2-2

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   collect biosurvey data focus on macroinvertebrates.
   (This may be due to the emphasis placed on mac-
   roinvertebrates for community-level evaluations in
   the 1976 Basic Monitoring  Programs Guidance.)
   •  Many States already have background macroin-
      vertebrate data.
   •  Most State water quality agencies have more
      expertise in aquatic entomology than in
      ichthyology.

Advantages of Using Fish

1.  Fish are good indicators of long-term (several
   years) effects and broad  habitat conditions because
   they are relatively long-lived and mobile (Karr et
   al.  1986).
2.  Fish communities generally include a range of spe-
   cies that represent a variety of trophic levels
   (omnivores, herbivores, insectivores, planktivores,
   piscivores).  They tend to integrate effects of lower
   trophic levels; thus,  fish community structure is
   reflective of integrated environmental health.
3.  Fish are at the top of the aquatic food chain and
   are consumed by humans,  making them important
   subjects in assessing contamination.
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.
   •  Environmental requirements of common fish are
      comparatively well known.
   •  Life history information  is extensive for most
      species.
   •  Information on fish distributions is commonly
      available.
5.  Aquatic life uses (water quality, standards) are typi-
   cally characterized in terms of fisheries (coldwater,
   cool water, warmwater, sport, forage).
   •  Monitoring fish communities provides direct
      evaluation of  "fishability", which emphasizes the
      importance of fish to anglers and commercial
      fishermen.
6.  Fish account for nearly half of the endangered ver-
   tebrate species and subspecies in the United States.

Advantages of Using Algae

1.  Algae generally  have rapid reproduction rates and
   very short life cycles, making them valuable indi-
   cators of short-term  impacts.
2.  As primary producers, algae are most directly
   affected by physical  and chemical  factors.
3.  Sampling is easy, inexpensive, requires few people,
   and creates  minimal impact to resident biota.
4.  Relatively standard methods exist for evaluation of
   functional and non-taxonomic structural (biomass,
   chlorophyll  measurements)  characteristics  of algal
   communities.
5.  Algal communities are sensitive to some pollutants
   which may  not visibly  affect other aquatic commu-
   nities, or may only affect other communities at
   higher concentrations (i.e.,  herbicides).
          2.3  STATION SITING
   RBP I, RBP II, RBP III, and  RBP V include the
collection of biological samples to assess the biotic
integrity of a given site. To meaningfully evaluate bio-
logical condition,  sampling locations must be carefully
selected to ensure generally comparable habitat at
each station. Unless basically comparable physical
habitat is sampled at all stations,  community differ-
ences attributable  to a degraded habitat will be diffi-
cult to separate from those resulting from water
quality  degradation. Availability of habitats at each
sampling location  can be established during prelim-
inary reconnaissance (such as RBP I). In evaluations
where several stations on a waterbody will be com-
pared, 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.
   Locally modified sites, such as small impound-
ments and bridge  areas, should be avoided unless data
are needed to assess their effects. 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).
   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 control  is generally
thought to be most representative of "best attainable"
conditions for  a particular waterbody. However,
regional reference stations may also be desirable to
allow evaluation of conditions on a larger scale.
Where feasible, effects should  be bracketed  by  estab-
lishing 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.
                                                     2-3

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          2.4 IMPORTANCE OF
        HABITAT ASSESSMENT
   The procedure for assessing habitat quality
presented in this document (Section 5.2) is 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 sur-
rounding land. All of the habitat parameters  evaluated
are related to  overall aquatic life use and are  a poten-
tial source of  limitation to the aquatic  biota.
    Habitat, as affected by instream and surrounding
topographical  features, is a major determinant of
aquatic community potential.  Both the quality and
quantity of available habitat affect the structure  and
composition of resident biological communities.
Effects of such features can be  minimized by sampling
similar habitats at all stations being compared. How-
ever, when  all stations are not physically comparable,
habitat characterization is particularly  important  for
proper interpretation of biosurvey results.
    Where habitat quality is similar, detected impacts
can  be attributed to water quality factors. However,
where habitat quality differs substantially from refer-
ence 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 rapid
bioassessment protocols  provides an effective means of
evaluating  and documenting habitat quality at each
biosurvey station.
  2.5 THE ECOREGION CONCEPT

    Innate regional differences exist in forests, agricul-
tural  potential, wetlands,  and waterbodies. These
regional differences  have  been mapped by Bailey
(1976); ~USDA Soil Conservation Service  (1981),
Energy,  Mines and Resources Canada (1986), and
Omernik (1987).  All four maps were developed from
examination of several mapped  land variables.  It is
assumed that waterbodies reflect the lands they drain
and that similar lands should produce similar water-
bodies. This ecoregional approach provides much
more robust and ecologically-meaningful  regional
maps than could be attained by mapping  a single vari-
able.  For example, hydrologic  unit maps  are useful  for
mapping drainage patterns, but have limited value for
explaining the substantial changes that occur in water
quality and biota independent of stream size and river
basin. Recognition of these changes stimulated
Warren's (1979) work, and Ohio's and Arkansas'
development of ecoregional standards.
   Omernik (1987) provides 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, potential
natural vegetation, and land use, which  vary across
the country. Two major applications grew out of the
regional approach. The first was the use of a rela-
tively small number of minimally-impacted regional
reference sites to assess feasible but protective biologi-
cal goals for an  entire region (Hughes et al. 1986).
The second was the use of regions as a statistical
framework for stratified random sampling of lakes in
a national survey of the effects of acid deposition
(Linthurst et al.  1986, Landers et al. 1987). These two
site selection methods offer ecologically and statisti-
cally valid means to establish baseline conditions and
assess water quality in entire regions by monitoring a
relatively  small number of sites.
   Geographic patterns of similarity among
ecosystems can be grouped into ecoregions. Naturally
occurring biotic assemblages, as components of the
ecosystem, would be expected to differ among
ecoregions but be relatively  similar within  a given
ecoregion. The ecoregion concept thus provides  a
geographic framework for more efficient management
of aquatic ecosystems and their components (Hughes
et al.  1986, Hughes 1985, and Hughes and Larsen
1988).  For example, studies in  Ohio (Larsen et al.
1986),  Arkansas (Rohm et al.  1987), and Oregon
(Hughes et al. 1987, Whittier et al.  1988) have shown
that distributional patterns of fish communities coin-
cide with the States' ecoregions as defined a priori by
Omernik (1987). This, in turn,  implies that similar
water quality standards, criteria, and monitoring
strategies are likely to be valid throughout  a given
ecoregion, but should be tailored to accommodate  the
innate differences among ecoregions (Ohio EPA
1987b).  Figure 2.5-1 shows the 76 ecoregions devel-
oped by Omernik (1987) for the conterminous United
States.
   Macroinvertebrate  communities reflect habitat
differences on a smaller scale than  fish,  and may be
better suited for site-specific assessments. Within an
ecoregion (Omernik  1987), additional qualifiers such
as stream size, hydrologic regime,  and riparian vegeta-
tion need to be considered.  In  addition, streams or
stream segments may  represent characteristics  atypical
for that particular ecoregion. For instance, a given
                                                     2-4

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stream segment may be wooded (deciduous or conifer-
ous) or open, within a perennial or intermittent flow
regime, and represent a particular stream size  (Figure
2.5-2). Individual descriptors may not apply to all
ecoregions, nor will all conditions (i.e., deciduous,
coniferous, open) be present in all stream sizes.
   The final  rapid bioassessment guidance should be
generally applicable to all ecoregions of the  United
States, although specific elements and evaluation
criteria may require modification for particular
ecoregions. When rapid bioassessment protocols  are
used to assess impact sources (upstream-downstream
studies), reference criteria may  not be as important if
an unimpacted site-specific control station can  be sam-
pled.  However, when a synoptic ("snapshot") survey
is being conducted or an appropriate control does not
exist in the immediate study area, use of idealized
criteria may be the  only means of discerning use
impairment or assessing impact.
   Each agency will need to evaluate the generic
criteria suggested in this document for inclusion  into
specific programs. To this end,  the application of the
ecoregion concept versus the site-specific control
approach will need  to be evaluated by each agency. It
is likely that  additional investigation will be  needed
to: delineate  areas that differ significantly in their
innate biological potential;  locate reference sites
within each ecoregion that  fully support aquatic  life
uses;  and develop biological criteria (e.g., define
optimal values for the metrics recommended) using
data generated from one of the  higher level protocols.
       2.6 DATA MANAGEMENT
              AND ANALYSIS


       2.6.1 Integration into BIOS

   The U.S.  Environmental  Protection Agency (EPA)
has developed a biological data management system
known as BIOS. BIOS provides a centralized system
for storage of biological data in addition to analytical
tools for data analysis. The field survey file compo-
nent of BIOS  provides a  means of storing,  retrieving,
and analyzing biosurvey data, and will process data
on the distribution, abundance, and physical condition
of aquatic organisms, as  well as descriptions of their
habitats. Data stored in BIOS become part  of a com-
prehensive database that can  be used as a reference, to
refine analysis techniques, or to define ecological
requirements for aquatic  populations.  Data  from the
rapid bioassessment protocols can be  readily managed
with the BIOS field survey file using  header informa-
tion presented in Figure 2.6-1 to identify sampling
stations.
   Habitat information and physical characterization
information may also be stored in the field survey file
with abundance data. Parameters available in the field
survey file can be used to  store some of the environ-
mental characteristics associated with the sampling
event, including physical characteristics,  water quality,
and habitat assessment. Physical/chemical parameters
with discrete  values may be stored in the field survey
file or the water quality file of STORET, under the
same station description. Such parameters include
stream depth, velocity,  and substrate characteristics, as
well as many other parameters. The system will also
allow storage of other pertinent station or sample
information in the comments section.

 2.6.2  Computerizing  Field  Data for
       Calculation of  the Metrics

   Entering data into a computer system can provide
a substantial time savings.  An additional advantage to
computerization is analysis documentation, which is
an important  component for a QA/QC plan. An
agency conducting rapid bioassessment programs can
choose an existing system  within their agency or
utilize the BIOS system developed as a national
database  system.
   The field survey file of BIOS can calculate several
metrics used in the  RBPs.  Metric values that may cur-
rently be calculated in a BIOS PGM = TAXATABLE
retrieval  report include taxa richness, EFT Index, and
percent contribution of the dominant taxon. Other
metrics are planned as future additions to the field
survey file. Additional metrics can be calculated using
SAS,  which is easily accessible to the file. BIOS  may
also be used  to create a machine-readable file for use
as input  to either a  user-written program or to an
external  analytical software package (SPSS, BMDP,
dBase III).
     2.7 BENTHIC COMMUNITY
            CONSIDERATIONS


            2.7.1  Seasonality for
            Benthic  Collections

   Rapid bioassessment is based on evaluation of rela-
tively few samples al a site.  Seasonality  is particularly
important when only a few collection sites are
involved. The  intent of the benthic rapid bioassess-
                                                    2-6

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                                  Ecoregion
                                 A, B, C, etc.
                                 Surrounding
                                   Land Use
                                  Evaluation
Figure 2.5-2. Flowchart illustrating potential delineation of reference sites within an ecoregion.
                                       2-7

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00
                              Waterbody Name.



                              Reach/Milepoint _



                              County	
                              Station Number.



                              Date	
                              Hydrologic Unit Code



                              Reason for Survey	
 State.
Time.
                            Location.
Latitude/Longitude.



Aquatic Ecoregion _
Investigators._



Agency	
                            Form Completed by
                                        Figure 2.6-1.  Header information used for documentation and identification for sampling stations.

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ment  is to evaluate overall biological condition,
optimizing the use of the benthic community's capac-
ity to reflect integrated environmental effects over
time.  Ideally, the optimal biological sampling season
will correspond to recruitment cycles of the inver-
tebrates. Maximum information for a benthic commu-
nity is obtained when most benthic macroinvertebrates
are within a size  range (later instars) retained during
standard sieving and sorting, and can be identified
with the most confidence.
   Reproductive periods and different life stages of
aquatic insects are related to the abundance of particu-
lar food supplies  (Cummins and Klug 1979). Peak
emergence and reproduction typically occur in the
spring and fell, although onset and duration vary
somewhat across  the United States. During peak
reproduction, approximately 80 percent of the macro-
invertebrates will be too small to be captured in suffi-
cient  numbers to  accurately characterize the
community. Additionally, food source requirements for
early  instars are different from those for later instars.
Therefore, the biologically  optimal sampling season
would occur when the habitat is utilized most heavily
by later instars and the food resource has stabilized to
support a balanced indigenous community.
    Field collections scheduled to  correspond to sea-
sonal recruitment cycles of invertebrates will provide
the optimal biological sampling period. However,
sampling during  these optimal biological periods may
not be  logistically feasible due to adverse weather
conditions, manpower availability, scheduling con-
straints, or other factors. Additionally, an agency may
be required to perform biological sampling during
periods of greatest environmental  stress such as low
flow/high temperature periods for point-source dis-
charges or high flow/runoff periods for nonpoirit-
source discharges.  Although an estimate of benthic
community structure during optimal biological condi-
tions is expected to reflect effects of, or recovery
from, environmental stress periods (Ohio EPA 1987a),
assessment of worst-case conditions may be  needed
under certain permitting regulations, or as a follow-up
to sampling during biologically optimal periods,
where impairment is detected.
    Optimal biological conditions for sampling vary
with  climate. Seven major climatological regions are
represented within the United States (Figure 2.7-1).
Temperature and/or rainfall are the principal  factors
influencing optimal biological conditions in each
climatological region. Several ecoregions are
represented within each of these  climatological
regions. Some scaling of the optimal collection period
will be necessary, depending on the elevation of the
site and the habitat type.
2.7.2  Benthic Sampling Methodology

2.7.2.1  Natural and Artificial Substrates

   The benthic RBPs employ direct sampling of natu-
ral substrates. Because routine evaluation of a large
number of sites is a primary objective of the RBPs,
artificial  substrates were eliminated from consideration
due to time required for both placement and retrieval,
and the amount of exposure time required for coloni-
zation. However,  where conditions are inappropriate
for the collection of natural substrate samples, artifi-
cial substrates may be an option. Artificial substrates
may be useful in  situations such as large rivers, where
an impact is attributable to physical alteration and
channelization or chemical effects. Artificial substrates
may be used to separate the two impact sources.
   Advantages and disadvantages of artificial sub-
strates (Cairns 1982) relative to the use of natural sub-
strates are presented below.

Advantages of Sampling With Artificial Substrates

 1.  Artificial substrates allow sample collection in
    locations that are typically difficult to sample
    effectively (e.g., bedrock, boulder, or shifting sub-
    strates; deep or high velocity water).
2.  As a  "passive" sample collection device, artificial
    substrates permit standardized sampling by
   eliminating subjectivity in sample collection tech-
    nique. Direct sampling of natural substrate requires
   similar effort  and degree of efficiency for the col-
   lection of each sample. Use of artificial substrates
    requires standardization of setting and retrieval;
   however, colonization provides the actual sampling
   mechanism.
3. Confounding effects of habitat differences are
   minimized by providing a standardized micro-
   habitat.  Microhabitat standardization may promote
   selectivity for  specific organisms if the artificial
   substrate provides a different  microhabitat than that
   naturally available at a site (see Disadvantage 2).
   Most artificial substrates, by design, select for the
   Scraper and Filtering Collector communities,
   which are the  macroinvertebrate communities
   emphasized in this document. However, in  some
   situations, accumulation of debris may cause a pre-
   dominance of Collector-Gatherers (Hilsenhoff, per-
   sonal communication).
4. Sampling variability is decreased due to a reduc-
   tion in  microhabitat patchiness, improving the
   potential for spatial and temporal similarity among
   samples.
5. Sample collection using artificial substrates may
                                                     2-9

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                                                                                                                                          Boston, Mass.
                                                                                                                                   New York, N.Y.
= Mediterranean



= Humid Continental



= Humid Subtropical



= Highlands
= Steppe



= Desert



= Marine West Coast
                             Figure 2.7-1. Classification of U.S. climatological regions. (Taken from Gabler et al. 1976.)

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   require less skill and training than direct sampling
   of natural substrates.  Depending on the type of
   artificial  substrate  used, properly trained techni-
   cians could place and retrieve the substrates.
   However, an experienced specialist should be
   responsible for the selection of habitats and sample
   sites.

Disadvantages of Sampling
With Artificial Substrates
1.  Two trips (one to set and  one to retrieve) are
   required  for each artificial substrate  sample; only
   one trip  is necessary for direct sampling of the
   natural substrate.  Artificial substrates require a
   long (8-week average) exposure period for coloni-
   zation. This decreases their utility for certain rapid
   biological assessments.
2.  Samples  may not be fully representative of the
   benthic community at a station if the artificial sub-
   strate offers different microhabitats than those
   available in the natural substrate.  Artificial sub-
   strates often selectively sample certain taxa, mis-
   representing relative abundances of these taxa in
   the natural substrate.  Artificial substrate samples
   would thus indicate colonization potential rather
   than  the  resident community structure. This could
   be advantageous if a study is designed to isolate
   water quality effects from substrate and other
   microhabitat effects.  Where habitat quality  is a
   limiting factor, artificial substrates could be used to
   discriminate between physical  and chemical effects
   and assess a site's  potential to support aquatic life
   on the basis of water quality alone.
3.  Sampler  loss or perturbation commonly occurs due
   to sedimentation,  extremely high or low flows, or
   vandalism during the relatively long  (at least
   several weeks)  exposure period required for
   colonization.
4.  Depending on the  configuration of the artificial
   substrate used, transport and storage can be diffi-
   cult. The number of artificial  substrate samplers
   required  for sample  collection increases such
   inconvenience.
 2.7.2.2  Single and Multiple Habitat Sampling

    A central  issue in the development of the rapid
 bioassessment protocols has been whether sampling
 all available habitats is necessary to evaluate biological
 integrity at a  site, or if sampling only selected habitats
 will provide a sufficient characterization. The pilot
 study in North Carolina (see Section 6.4) addressed
 this issue and indicated that the  riffle community and
the multihabitat assemblage responded similarly to
differences among stations.  For example, under stress,
taxa richness was reduced by the same proportion in
both the riffle community and  the multihabitat assem-
blage at a given station. These responses suggest that
either the riffle community  or  the multihabitat assem-
blage will give a good assessment of biotic integrity
but assessing both may be redundant.
   The sampling of a single habitat type (e.g., riffle/
run) is  intended to  limit the variability inherent in
sampling natural substrates. Kicknet samples are used
in the RBPs because they have been shown to provide
good statistical replication (Pollard 1981). However,
some streams  lack the cobble substrate (riffle/run) to
support the periphyton-based benthic community
emphasized in the RBPs. In this case, an alternate
habitat(s) will need to be sampled. Some State agen-
cies, such as North Carolina DEM, have been suc-
cessful  in using a multihabitat sampling  approach, and
advocate this technique  as being more appropriate in
North Carolina than simply sampling the riffle/run
habitat.
   Discussions at the 1987 Biocriteria Workshop (U.S.
EPA 1988) indicated strong  support for multihabitat
sampling where time and resources permit and the
particular region or specific study emphasizes non-
Scraper communities. It was generally agreed, how-
ever, that samples from various habitats should be
processed and analyzed separately. Data can always  be
aggregated after individual samples are analyzed and
tabulated, but potentially important comparisons
among habitats are  lost if samples are composited.

2.7.2.3 Sampling Coarse  Participate
Organic Material (CPOM)

   In addition  to sampling the riffle habitat, the ben-
thic  RBPs recommend that  a Coarse Particulate
Organic Material (CPOM) sample also be collected
(Sections 6.2.1.1 and 6.3.1.1). In lotic systems, CPOM
generally exists in -the form of plant debris (leaves,
needles, twigs, bark) which accumulates in deposi-
tional areas. The rationale for collecting the CPOM
samples is that, as  a group, Shredders should be par-
ticularly affected by toxic pollutants that often adsorb
to CPOM (Cummins 1987, personal communication).
Toxicants adsorbed to CPOM affect Shredders directly
through ingestion and indirectly by killing attached
microbes that "prepare" CPOM for Shredder con-
sumption. In a study by Newman et al.  (1987), amphi-
pod  Shredders  colonizing litter bags were significantly
reduced in numbers by  230 5g/L total residual
chlorine.
   The CPOM  sample  is processed separately and
                                                     2-11

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the organisms are identified as Shredders or Non-
Shredders (Sections 6.2.1.1 and 6.3.1.1). Taxonomic
identifications are not necessary in that most aquatic
insects can be classified to Functional Feeding Group
on the basis of morphological and behavioral features
using the procedures in Cummins and Wilzbach
(1985). These counts are used to calculate the ratio of
Shredders to  the total number of individuals collected,
one of the eight metrics used in the final biosurvey
analysis.

    2.7.3  Benthic  Sample Processing
              and Enumeration

   One of the underlying goals of this guidance is to
promote consistency in the conduct of rapid bioassess-
rnents. In RBP II, consistent sample  handling is very
important because relatively detailed  comparisons are  •
made among stations and sites. The RBP n 100-count
subsampling  procedure is adapted from Hilsenhoff
(1987b) and is performed in the field. The level of
 effort and subsampling process used for RBP III is
 similar to RBP II,  except subsampling is conducted in
 the laboratory. This laboratory subsampling procedure
 provides a more standard unit of effort.
    Much of the useful information regarding assess-
 ment of biological  condition can be obtained from a
 relatively small subsample, such as the 100-count sub-
 sample recommended  in this document (see Section
 6.4).  However, an agency may be willing to expend
 additional time to attain a higher degree of resolution.
 In this case, a 200- or 300-count subsample may be
 selected.  Some agencies may wish to process the   /
 entire sample for analysis.

       2.7.4  Benthic Environmental
       Tolerance Characterizations

    Assessment of biological condition using the ben-
thic protocols presented in this document is based on
the calculation of several metrics. Certain metrics rely
on classification of benthic taxa according to their
relative sensitivity to pollution. This  approach  reflects
the longstanding indicator species concept, with sensi-
tivity primarily related to responses of species to
organic pollution. Responses to toxicants are also
incorporated  into the tolerance characterizations,  but
to a lesser extent.  Evaluation of toxic effects is
addressed primarily at the Functional Feeding  Group
level.
    The specific tolerance characterizations used in the
RBPs  were obtained from Hilsenhoff (1987b, 1988).
Hilsenhoff s species level tolerance characterization
system was selected due to its extensive use across the
country. A more recently developed  family level sys-
tem (Hilsenhoff 1988) has not been used extensively,
but was based on  Hilsenhoffs original widely
accepted index.  Several other general tolerance clas-
sification systems  are fairly  well established,  generally
applicable, and may also be used as guidelines. Some
of these are listed in Appendix C. Additional biotic
indices are also listed in U.S. EPA  1983. However,
types of pollution  and causes of impairment will differ
regionally, and the meaning of  "pollution tolerance"
may vary among regions. Therefore, optimal
implementation of the tolerance characterization
approach requires that each State agency refine estab-
lished tolerance classification systems for their own
use. Winget and Mangum (1979) have developed a
tolerance classification based on nonpoint-source
effects  (Biotic Condition Index). This classification
may prove useful  as a substitute for Hilsenhoffs when
evaluating nonpoint-source problems.
         2.8  FISH COMMUNITY
            CONSIDERATIONS

 2X1 Seasonality for Fish Collections

    Seasonal changes in the relative abundances of the
 fish community primarily occur during reproductive
 periods and (for some species) the spring and fall
 migratory periods. However, because larval fish sam-
 pling is not recommended in this  protocol,  reproduc-
 tive period changes in relative abundance are not of
 primary importance.
    Generally,  the preferred sampling season is mid to
 late summer, when stream and river  flows are moder-
 ate to low, and less variable than  during other sea-
 sons. 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 Environmental Protection Agency (Rankin
 1987, personal communication) confirmed that few
 fishes in perennial streams migrate long distances.
 Hill and Grossman (1987) found that the three domi-
 nant fish species in a North Carolina stream had
 home ranges of 13 to 19 m over a period of 18
 months. Ross  et a!. (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.
                                                   2-12

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   2.8.2 Fish Sampling Methodology

2.8.2.1  Use of Electrofishing, Seining,
and Rotenoning

   Although various gear types are routinely used to
sample fish, electrofishers, seines, and rotenone are
the most commonly used collection methods  in fresh-
water habitats. Each method has advantages and disad-
vantages  (Nielsen and Johnson 1983; Hendricks et al.
1980).  However, electrofishing is  recommended for
most fish field surveys because of its greater  applica-
bility and efficiency. Local conditions may require
consideration of seining and rotenoning as optional
collection methods. Advantages and disadvantages  of
each gear type are presented below.

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 ichthyo-
   cides) (Hendricks et al. 1980).
3.  Electrofishing is less selective than  seining
   (although it is selective towards size and species)
   (Hendricks et al.,  1980). (See disadvantage
   number 2).
4.  If properly used, adverse effects on fish are
   minimized.
5.  Electrofishing is appropriate in a variety of
   habitats.

Disadvantages of Electrofishing

1.  Sampling efficiency is affected  by turbidity and
   conductivity.
2.  Although less selective than seining, electrofishing
   is size and species selective. Effects of electrofish-
   ing increase with body size. Species specific
   behavioral and anatomical differences also deter-
   mine  vulnerability to electroshocking (Reynolds
   1983).
3.  Electrofishing is a hazardous operation that can
   injure field-personnel  if proper safety procedures
   are  ignored.

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.

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 gear (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 with little cover.
4. Standardization of unit of effort  to ensure data
   comparability  is difficult.

Advantages of Rotenoning

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 censusing of the fish popu-
   lation than seining or electrofishing.

Disadvantages of Rotenoning

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 tempera-
   ture,  light, dissolved oxygen, alkalinity, and tur-
   bidity (Hendricks  et al.  1980).
4. Rotenoning typically has a high environmental
   impact; concentration miscalculations can  produce
   substantial fish kills downstream of the study site.

2£.2.2 Sampling  Representative Habitat

   The sampling approach  advocated  in fish RBP V
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  site, and a  realistic
sample of fish likely  to be encountered in  the water-
body. When  sampling large streams, rivers, or water-
bodies with complex  habitats, a complete inventory of
the entire reach is not necessary for the level of
assessment used  in RBP V. 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
                                                   2-13

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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. Sam-
pling 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 RBP V.

  2.8.3 Fish Sample Processing and
                 Enumeration

   To  ensure data comparability for assessing biologi-
cal condition with fish RBP V,  sample processing and
species enumeration must be standardized.
   Processing of the fish biosurvey sample includes
identification of all individuals to species,  weighing (if
biomass data are desired), and recording 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.) The
data from the counted and weighed subsample is
extrapolated for the total. Ohio EPA  (1987a) found that
subsampling reduced potential error and made the
extra time required for weighing insignificant. Proce-
dural details for subsampling are presented in Ohio
EPA 1987c. Determination of trophic guild designation
is also necessary for some FBI metrics.

 2X4  Fish Environmental Tolerance
             Characterizations

   Use of the IBI in fish RBP V requires classifica-
tion of fish species in terms of environmental toler-
ance. Responses of individual species to pollution will
vary regionally and according to the type  of pollutant.
Tolerance characterizations of selected midwestern and
northwestern  fish species are presented in Appendix
D. Effective use of the tolerance characterization
approach requires an appropriate regional  tolerance
characterization system. Regional modifications or
substitutions may be  based upon regional  fish refer-
ences,  historical distribution records,  objective assess-
ment of a large statewide database, and toxicological
test data. IBI  tests in the southeastern  and south-
western United States, and its widespread use by
water resource agencies may result in  additional
modifications. Past modifications have occurred (Sec-
tion 7.2.2.1, Miller et al. 1988a) without changing the
IBI's basic theoretical foundations.
                                                    2-14

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                     3.  OVERVIEW  OF  PROTOCOLS AND
                          SUMMARY  OF  COMPONENTS
   The bioassessment protocols presented in this
document provide guidance to those agencies not
presently using biosurveys as investigative tools or
who are seeking alternatives to their present methods.
Agencies with successful  biological monitoring pro-
grams are encouraged to continue their programs  and
provide further leadership and guidance in the use of
bioassessment.  The five separate protocols presented
below reflect different levels of effort and expertise
and focus on different objectives.
        3.1 SUMMARY OF THE
               PROTOCOLS
   A summary of the key features of the five pro-
tocols is presented in Table  3.1-1.  The first and fourth
protocols are subjective because an investigator or
agency may conduct any level of investigation deemed
necessary. The presence or  absence of impairment
using Rapid Bioassessment Protocols (RBPs) I and IV
is supported by a limited analysis of the biological
communities. Benthic  RBP I and  fish RBP IV are
used as screening or reconnaissance techniques for
discerning biological impairment. Benthic RBPs II and
III, and fish RBP V are progressively more rigorous
and are intended to provide more objective and repro-
ducible evaluations than RBPs I and IV.  RBPs II, III,
and V  are designed to be semi-quantitative and utilize
an integrated analysis  technique to provide continuity
in the evaluation of impairment among sites and sea-
sons. The primary difference between RBPs II, III
and V  is the level of taxonomic resolution (i.e.,
family  level vs. genus/species level identification)
necessary to perform an assessment. RBPs III and V
require more time and expertise than RBP II, but are
better able to discriminate degrees of impairment.
          3.2  OBJECTIVES OF
            THE PROTOCOLS
    As presented in Figure 3.2-1, selection of the
 appropriate bioassessment approach depends on the
 objectives of the study. RBPs I and IV provide a
screening mechanism for identification of biological
impairment; they are not intended to quantify the
degree of impairment nor provide definitive data that
would be used to establish a cause-and-effect. RBPs I
and IV allow a cursory assessment incorporating the
cost and time efficiencies necessary to evaluate a large
number of sites. RBPs I and IV are used primarily to
identify major water quality problems as an aid in
planning and developing management strategies.
   The information derived from benthic RBP II pro-
vides  a basis for ranking sites as severely or moder-
ately impaired. This classification can then be used to
focus  additional study or regulatory action. RBP II
can also be used as a screening technique in lieu of
benthic RBP I. Like RBP I, RBP II  is designed to
enable agencies to evaluate a large number of sites
with relatively limited time and effort. However, the
concept of a documented procedure for collections,
inherent in RBP II and intended to promote a consis-
tent level of effort, allows for better comparison
among sites.
   The primary objective of benthic RBP III  and fish
RBP V is to provide a consistent, well-documented
biological assessment. Repeatable results provide a
basis  for  comparison among sites over time (trend
monitoring). The ability to discriminate the level of
impairment among sites is enhanced by performing
taxonomic identifications to the lowest practical level,
thereby providing information on population as well as
community level effects. RBPs III and V can  be used
to rank sites according to impairment in lieu of
RBP II, but will also establish a basis for trend
monitoring over a period of time. RBPs III and V
place still greater emphasis on consistency in unit
effort and documentation.
    3.3 LEVEL OF EFFORT AND
    INVESTIGATOR EXPERTISE

   The level of effort required for RBP I is estimated
to be  1 to 2 hours per station, excluding travel time.
The effort consists of habitat assessment, physico-
chemical measurements, and biological collections and
observations. All of this effort is expended in the
field.  An additional 0.5 to 1 hour of data analysis and
                                                 3-1

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                                       TABLE  3.1-1   COMPARISON  OF RAPID BIOASSESSMENT  PROTOCOLS
                          Protocol I
                                                  Protocol II
                                                                           Protocol III
                                                                                                     Protocol IV
                                                                                                                              Protocol V
Objectives
Level of Effort
(per station}
Experience
Required
Minimal Skill Mix
Habitat Assessment
Water Quality and
Phys/Chen
Determine whether
biological inpair-
aent exists
Determine if further
investigation is
needed
Field—1 to 2 hours/
person (1 person)
Lab—None
Data—0.5 to 1 hour
(1 person)
Budget—Total of 1.5
to 3 work hours

High level of pro-
fessional impact
assessment
experience
Knowledge of benthic
invertebrate ecology

Biologist
Characterize and
rate substrate/
instream cover,
channel morphology,
and riparian/bank
structure

Measure conventional
water quality
parameters
Examine physical
characteristics
.  Assess  biological
  impairment
.  Provide infornation
  for  ranking sites
.  Prioritize sites  for
  further assessment
  and/or  testing
  (toxicity, chenical)
Field—1.5 to 2.5
hours/person (2 per-
sons )
Lab—None

Data — 2 to 4 hours
(1 person)
Budget—Total of 5
to 9 work hours

Professional impact
assessment experience
Knowledge of benthic
ecology and taxonomy
  Biologist  and
  technician

  Characterize and  rate
  substrate/instream
  cover,  channel
  morphology, and
  riparian/bank
  structure

  Measure conventional
  water  quality
  parameters
  Examine physical
  characteristics
                         Assess biological
                         impai rment
                         Establish basis for
                         trend monitoring
                         Prioritize for further
                         assessment and/or
                         testing (toxicity,
                         chenical)
Field—1 to 2 hours/
person (2 persons)
Lab—2 to 3 hours
(1 person)

Data—1 to 3 hours
(1 person)
Budget—Total of 5
to 10-work hours

Professional impact
assessment experience
Knowledge of benthic
ecology and taxonomy
                         Biologist and
                         technician

                         Characterize and rate
                         substrate/instream
                         cover, channel
                         morphology,  and
                         r ipa r ian/bank
                         structure

                         Measure conventional
                         water quality
                         parameters
                         Examine physical
                         characteristics
                          Determine whether
                          biological impairment
                          exists
                          Determine if further
                          investigation is
                          needed
                                                     Field—None

                                                     Lab—None
Data — 3 hours
(1 person)
Budget—Total- of
3 work hours

Survey design
experience
Knowledge in broad
patterns of fish
species distribution
and abundance

Biologist
                          Characterise and ratt
                          substrate/instream
                          cover, channel
                          morphology,  and
                          riparian/bank
                          structure

                          Measure conventional
                          water quality
                          parameters
                          Examine physical
                          characteristics
Assess biological
impa i rment
Establish basis for
trend aonitoring
Provide information
for ranking sites
Prioritize for
further assessment
and/or testing
(toxicity, chemical)

Field—1-5 hours/
pe rson
(2 persons minimum)
Lab—None

Data —1-2 hours
(1 person)
Budget—Total of 3 to
17 work hours

Professional impact
assessment experience
Knowledge in the use
of the IBI and 1KB
Biologist and
technician(s)

Characterise and rati
substrate/instream
cover, channel mor-
phology, and
riparian/bank
structure

Measure conventional
water quality
parameters
Examine physical
characteristics

-------
                                                            TABLE  3.1-1   (Cont.)
                          Protocol I
                                                  Protocol II
                                                                           Protocol III
                                                                                                     Protocol IV
                                                                                                                              Protocol V
Biosurvey
Cursory examination
Determine relative
abundance of aacro-
benthos; field IDs
Exaaination focusing
on the riffle/run
coaaunity,
supplenented with a
CPOH saaple
100-organisa sub-
sample IDed in field
to faaily or order
level
Functional  Feeding
Group analysis of
riffle/run  and CPON
sample in the field
Examination focusing
on the riffle/run
coaaunity,
supplemented with a
CPOH sample
Collect riffle/run
benthos;  collect CPON
saaple, deteraine
Shredder  abundance
Preserve  riffle/run
sample, return to lab,
do 100-organism sub-
sample, IDs to species
level and Functional
Feeding Group analysis
Questionnaire survey
Survey ecoregional
reference reaches and
randomly selected
reaches
Examination with
sampling of all major
habitats and cover
types
Collect fish, note
condition, ID to
species level in
the field
Preserve voucher col-
lection for deposit
in museum
Analysis
Conclusion
Minimal; deteraine
presence or absence
of iapairment
                      Determine if iapair-
                      ment exists
                      Indicate generic

                      (habitat, organic
                      enr ichaent,
                      toxicity|
Integrated assessaent
of metrics measuring
various components of
family level com-
munity structure
                        Characterise
                        conditions as no
                        impairment, moderate
                        impairment, severe
                        impa i rment
                        Indicate generic
                        cause of iapairaent
                        (habitat, organic
                        enrichaent, toxicity)
Integrated assessment
of aetrics measuring
various components of
genus/species level
community structure
                         Evaluate site as no
                         impairment, slight
                         iapairaent, aoderate
                         impairment, severe
                         impairment
                         Indicate generic cause
                         of iapairaent
                         (habitat, organic
                         enrichaent, toxicity)
Summarize survey
responses to deter-
aine degree and
probable cause of
impairment
                          Deteraine if impair-
                          ment exists
                          Indicate generic
                          cause of iapairment
                          (habitat, water
                          quali ty)
Integrated assessment
of metrics measuring
various components of
species, family, and
trophic level com-
munity structure

Evaluate biological
integrity as
excellent, good,
fair, poor, very poor
Indicate generic
cause of impairment
(habitat, organic
enrichment, toxicity)

-------
                                               APPROACH
                                               Decide on
                                               Monitoring
                                               Objectives


SCREENING


-Limited Effort
-Impairment
Noted




r~
i

SITE RANKING




Level One I
1

-Focus on
Communities
-Three Levels
of Impairment
Detected




i
Level Two

1

-Focus on
Communities
and Populations
-Four Levels
of Impairment
Detected
              Figure 3.2-1.  Overview of the five bioassessment approaches and their primary objectives.
evaluation might be needed.  No laboratory analysis is
anticipated for RBP I. Only  a single experienced biol-
ogist is needed to conduct RBP I. However,  a second
person, for reasons of safety, quality control, or train-
ing, may be assigned by the  agency.
   The biologist conducting RBP I should have
professional impact assessment experience with knowl-
edge of benthic invertebrate  ecology. A HIGHLY
EXPERIENCED  INDIVIDUAL IS REQUIRED. The
accuracy of the assessment depends upon the biolo-
gist's professional integrity.
   The field  survey for RBP II can be completed  by
two persons in 1.5 to 2.5 workhours per person for
each station,  excluding travel time. The necessity for a
reconnaissance survey (similar to RBP I) is  somewhat
dependent on agency familiarity  with the site and pur-
pose of the investigation. Habitat assessment,  physico-
chemical  measurements, and biological  collections/
observations are  also required  in this protocol. All of
the sorting and identifications (family level)  are done
in the field to minimize laboratory time. RBP II
requires one field investigator experienced in impact
assessment, benthic  ecology, and taxonomy.  The sec-
ond field person can be a trained technician with a
biological background. Data analysis may take from  1
to 2 hours per station and should be performed by an
experienced biologist. Use of computers for data entry
and calculation of results provides optimal time effi-
ciency for data analysis. Hand calculation of results
may require an additional 1 to 2 hours per station for
data analysis.
   The field effort for RBP III can be done by two
biologists in 1 to 2 workhours per biologist  for each
station. RBP III is the most detailed of the benthic
protocols and provides a means of obtaining repeat-
able results; the sorting and identification tasks are
conducted in the laboratory. Laboratory processing is
estimated to take 2 to 3 hours per station for one biol-
ogist. Data analysis is expected to take an additional 1
to 3 hours per station for one biologist. One of the
field investigators must be experienced in impact
assessment of benthic communities. This investigator
should also perform the data analysis to provide con-
tinuity throughout the assessment process. The person
performing the laboratory analysis needs to be
experienced in taxonomy, but does not necessarily
need to perform the final assessment. Data analysis
can be expedited by computerized data entry for cal-
culation  of results. An additional  1  to 2 hours per sta-
tion may be required  if results are hand calculated. As
field and analysis procedures are  mastered, time effi-
ciencies  can be expected.
   The level of field effort for RBP IV requires 1 to
3 work hours per reach. This includes reach selec-
                                                     3-4

-------
tion, respondent identification,  mailing, questionnaire
completion, and response tabulation. No additional
field or laboratory work is required and no travel or
equipment are necessary. All effort is expended  in the
office and is independent of season or weather condi-
tions. The designer of the survey must be familiar
with survey design. Respondents should also  be  famil-
iar with broad patterns in species distribution and
abundance. The accuracy of statements depends  on the
respondent's professional knowledge and integrity.
Only one  respondent is needed per reach but some
confirmation from additional sources and methods is
advisable  for quality assurance.
   The level of effort for RBP  V includes  1 to 5
hours for  fish collection  and identification  (depending
upon habitat complexity  and  gear used), and  1 to 2
 hours for data analysis. Data analysis can be per-
 formed by one biologist.  The field effort requires a
 minimum of 2 persons; up to 5 people may be needed
 in wide shallow rivers and rivers  with complex
 habitats. Typically, using  this protocol, stream sites
 can be sampled and data  analyzed within a total of 5
. hours. At least one biologist should have professional
 impact assessment experience, with specific knowl-
 edge in the use of the Index of Biotic Integrity (IBI)
 and the Index of Well Being (IWB).  This biologist
 should be involved in all  phases of the protocol to
 improve continuity and to provide greater insight. The
 remaining crew members can be experienced techni-
 cians.  Sample quality can be assessed by replicate
 sampling and inter-crew sampling.
                                                     3-5

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            4.  QUALITY ASSURANCE/QUALITY CONTROL
   Effective quality assurance and quality control
(QA/QC) procedures and a clear delineation of
QA/QC responsibilities are essential to ensure the
utility of environmental monitoring data. The term
"quality control" refers to the routine application of
procedures for obtaining prescribed standards of per-
formance in the  monitoring and measurement process.
The term "quality assurance" includes the quality
control functions and involves a totally integrated pro-
gram for ensuring the reliability of monitoring and
measurement data.
   4.1 PROGRAM DESCRIPTION
   The U.S. EPA QA/QC program requires that all
EPA national program offices, EPA regional offices,
and EPA laboratories participate in a centrally
planned,  directed, and coordinated Agency-wide
QA/QC program. This requirement also applies to
efforts carried out by the States and interstate agencies
that are  supported by EPA through grants, contracts,
or other formalized agreements. The EPA QA pro-
gram is  based upon EPA order 5360.1, "Policy and
Program Requirements to Implement the Quality
Assurance Program" (U.S. EPA 1984a), which
describes the policy, objectives, and responsibilities of
all EPA  Program and Regional offices.
   Each office or laboratory that generates data under
EPA's QA/QC program must implement,  at a mini-
mum, the prescribed procedures to ensure that preci-
sion, accuracy, completeness, comparability, and
representativeness of data are known and documented.
 4.2  DATA QUALITY OBJECTIVES


   A full assessment of the data quality needed to
meet the intended use should be made prior to
specification of QA/QC  controls. The determination of
data quality is accomplished through the development
of data quality  objectives (DQOs). DQOs are qualita-
tive and quantitative statements developed by data
users to specify the quality of data needed to support
specific decisions or regulatory actions. Establishment
of DQOs involves interaction of decision-makers and
the technical staff.
   The process for developing DQOs includes a first
stage involving input by the decision-maker regarding
the information needed, reasons for the need, how the
information will be used, and specification of any
time and resource constraints. The second stage in
developing DQOs involves clarification of the specific
problem. Here, the technical staff and decision-maker
interact to establish a detailed specification of the
problem and any constraints imposed on data collec-
tion. The third stage involves developing alternative
approaches to data collection, selecting the approach
to be used, and establishing the final data quality
objectives. Once  the data collection approach and data
quality objectives have been established,  a clear
understanding of data quality will help ensure success-
ful study completion. U.S. EPA (1984b) describes the
process for developing DQOs in  more detail.
     4.3  QUALITY ASSURANCE
       PROGRAM PLANS AND
            PROJECT PLANS

   To provide adequate control and guidance, the
Agency's  QA program relies on the development and
implementation of two  QA documents: the QA Pro-
gram Plan and the QA Project Plan. These plans are
required of all recipients  of EPA grants and assistance
programs. Grant regulations, 40 CFR Part 30, require
submission of QA  Program Plans to EPA as a prior
condition of receiving an EPA grant. QA Project
Plans also must be developed according to an accept-
able schedule within the QA Program Plan. The QA
Program Plan (U.S. EPA 1980a) describes manage-
ment policies, organization, objectives, principles, and
general procedures that establish how data of known
and acceptable  quality  will be produced. The QA
Project Plan describes  and defines specific objectives,
network design, procedures, methods, and controls
that will be applied to  a specific project to ensure the
production of data of known and acceptable quality.
Two guidance documents are  available to assist  in
preparation of the QA Project Plan: a general gui-
dance document (U.S. EPA 1980b) and  a more
detailed guidance document that combines a work
plan with the QA Project Plan (U.S. EPA 1984c).
These documents also provide guidance on the use of
a short form for limited surveys.
                                                  4-1

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    4.4 EPA RESPONSIBILITIES


  EPA Headquarters is responsible for providing gui-
dance for developing Quality Assurance Program
Plans and Quality  Assurance Project Plans. This
includes  updates necessitated by new Agency require-
ments and additional technical guidance for the
Regional offices and States to develop sound  plans. In
addition, Headquarters is responsible for developing
guidance for inclusion of Data Quality Objectives in
Quality Assurance Project Plans and providing gui-
dance to the Regions on application of the DQO
development process.  In order to provide  on-going-
guidance, the Office of Water has formed a Water
Monitoring  Data Quality Objective Advisory  Group.
   EPA Regional offices are responsible for develop-
ing Quality  Assurance Program Plans and Quality
Assurance Project  Plans for the activities  that they
conduct. In  addition, they are responsible for ensuring
that States prepare QA Program Plans and Project
Plans in  conformance with grant  requirements speci-
fied in 40 CFR Part 30.  The Regions are  responsible
for developing DQO requirements compatible with
Headquarter's requirements and meeting the Regions'
specific needs. The Regions are also .responsible for
assisting the States in developing DQO  requirements
that meet State needs.
    4.5  IMPORTANCE OF  QA/QC
  FOR RAPID BIOASSESSMENTS
   Quality assurance and control (QA/QC) should be
a continuous process implemented throughout the
entire bioassessment program. All aspects of the
study, including field collection, habitat assessment,
lab processing, and data analysis are subject to
QA/QC procedures.  As with any scientific study,
quality must be assured before the results can  be
accepted. As described below, quality assurance is
accomplished-through  establishment of thorough inves-
tigator training, protocol guidelines, comprehensive
field and lab data documentation and management,
verification of data  reproducibility, and instrument
calibration.
   The protocols for rapid bioassessment presented in
this document can be modified to achieve specific
objectives. A different habitat assessment approach,
replicate sampling,  more intensive sample enumera-
tion, or modified analytical metrics may be  preferred
by a particular State over the methods suggested in the
RBPs.  Such refinements  can be accommodated,
provided they are clearly documented in an EPA
approved Quality Assurance Program and/or Project
Plan.

Training
   All personnel conducting assessments must be
trained in a consistent manner (preferably by the same
person) to ensure that the assessments are conducted
properly and to ensure standardization. At least one
investigator for each site should be a professional biol-
ogist trained and skilled in field aquatic sampling
methods and organism identification. Additionally, the
investigator should be familiar with the objectives of
each site investigation. Each agency should  have  a
designated QA/QC officer (or a person in charge of
the program) responsible for maintaining consistency
among investigators. At regularly scheduled intervals,
the QA/QC officer should visit selected overlap sites
and perform assessment techniques to use as a repli-
cate  of a previous assessment. Results from two sepa-
rate assessments conducted by two different teams can
determine if reproducible results  are being attained.
Quality control of taxonomic identifications can also
be evaluated in this way.

Standard Procedures
   It is the responsibility of each agency to define
precise methods and review inconsistencies  before the
assessment begins. Because RBPs I and IV  are
primarily subjective investigations, a specific level of
effort should be established prior to the actual field
investigation. Taxonomic identification is required at
various levels for RBP I (order level), RBP II  (family
level),  RBP III (genus and species level), and RBP V
(species level).  Field experience and taxonomic exper-
tise requirements for the particular level of assessment
performed must be  met. Any deviations from the pro-
tocol should be documented as to the reason for  devi-
ation, and corrective actions taken.
   Field validation, conducted at a frequency to  be
determined by each agency, should involve two proce-
dures:  (1) collection of replicate samples at  various
stations to check on the 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 to maintain objectivity
in the bioassessment.

Documentation
   The field data sheets should be filled out as com-
pletely and as  accurately as possible to provide a rec-
ord in  support of the survey and analysis conclusions.
Abbreviations  commonly used in documentation  (e.g.,
                                                    4-2

-------
scientific names) should be standardized to decrease
data manipulation errors. Field and laboratory data
sheets and  final reports should be filed.

Habitat Assessment

   Because the habitat characterization step of the
protocols is primarily a  subjective evaluation, final
conclusions are potentially subject to variability
among investigators. This limitation can be mini-
mized, however, by ensuring that each investigator is
appropriately trained in the evaluation technique and   •
periodic cross-checks are conducted among investiga-
tors  to promote consistency.

Benthic Collections

   The data developed during the benthic collection
effort is directly comparable to data developed at
other sites  because: (1) only similar habitats are  sam-
pled at each site, and (2) a uniform method (consis-
tent  unit of effort, 100-organism count) is used for
benthic data acquisition. To ensure that sampling
methods are applicable to a specific region, results
may be evaluated by comparing results obtained using
other sampling methods. To ensure reproducible data,
well characterized sites  should  be periodically resam-
pled by a variety of investigators. To document con-
sistency among field teams, selected sites should be
sampled simultaneously by several field teams.
Fish Collections

   To ensure fish field survey data is representative of
the fish assemblage at a particular site requires careful
regional analysis and station siting. Data comparability
is maintained by using similar collection methods and
sampling effort in waterbodies of similar size. Also,
where possible, major habitats (riffle, run, pool) are
sampled at each site, and the proportion of each  habi-
tat type sampled, should be comparable.
   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 differ-
ences exceeding 10 percent at minimally-impacted sites
and 15 percent at highly-impacted sites. IBI differ-
ences at the same site should not exceed 4 (Karr et al.
1986).
   Data reproducibility may be ensured by having a
variety of investigatprs periodically resample well
characterized sites. Investigator accuracy for use  of the
IBI and the IWB may be determined by having inves-
tigators evaluate a standard series of data sets or
preserved field collections.

Calibration of Instruments

   Instruments used for measuring water quality,  cur-
rent velocity, or any other  measurable parameters
should be calibrated with known  standards. All field
measurements  should  be accompanied by documenta-
tion of the type of instrument and the identification
number of the instrument  used.
                                                     4-3

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                          5.  HABITAT ASSESSMENT AND
                       PHYSICOCHEMICAL  PARAMETERS
    An evaluation of habitat quality is critical to any
 assessment of ecological integrity. The habitat quality
 evaluation can be accomplished by characterizing
 selected physicochemical parameters and 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.
               5.1 PHYSICAL
           CHARACTERISTICS
         AND WATER QUALITY

    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 5.1-1. The infor-
 mation  requested includes measurements made rou-
 tinely during biological surveys. This phase of the
 survey is broken into two sections: Physical Charac-
 terization and Water Quality  (Figure 5.1-1). These sec-
 tions are discussed separately below.

     5.1.1 Physical  Characterization

    Physical characterization parameters include esti-
 mations  of general land use and physical stream
 characteristics such as width, depth, flow, and sub-
. strate. 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 to
 presence of stream impacts. The information requested
 in the Physical Characterization section of the Field
 Data Sheet (Figure 5.1-1) is briefly discussed below.

 Predominant Surrounding Land Use: Observe the
 prevalent land-use  type in the vicinity (noting any
 other land uses in  the area which, although not  pre-
 dominant,  may potentially affect water quality).

 Local Watershed Erosion—The existing or potential
detachment of soil within the local watershed (the por-
tion of the watershed that drains directly into the
stream) and its movement into a stream is noted. Ero-
sion can be rated through visual observation of water-
shed and stream characteristics. (Note any turbidity
observed during water quality assessment below.)

Local Watershed Nonpoint-Source Pollution—This
item refers to problems and potential problems other
than  siltation. Nonpoint-source  pollution is defined as
diffuse agricultural and urban runoff.  Other com-
promising factors in a watershed that may affect water
quality are feedlots,  wetlands, septic systems, dams
and impoundments,  and/or mine seepage.

Estimated Stream Width (m): Estimate the distance
from shore to shore at a transect representative of  the
stream width  in the  area.

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.

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.

Velocity: Record an estimate of stream  velocity in  a
representative run area.

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 relat-
ing to  alteration of flow.

Channelized: Indicate whether or not the area around
the sampling  station is channelized.

Canopy Cover: Note the general proportion of open
to shaded area which best describes the amount of
cover at the sampling station.

Sediment Odors: Disturb sediment and note any
odors described (or include any other odors not listed)
                                                    5-1

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                                               PHYSICAL CBABACTERISATIOB/VATE8 QUALITY
                                                          FIELD DATA  SHEET
 PBT3ICAI. CBABJLCTERI1ATIOB

 BIPABIAB EOBE/IBSTBEAH FIXTURES

 Predominant Surrounding Land  Uaa:

 Poraet       riald/paiture     Agricultural       Raaidential       Co»ercial

 Local tratarahed Eroaion:   Ion*     Moderate     Heavy

 tool Waterehed BP3 Pollution:    Bo  evidence      Sone  Potential  Sourcea

 Eatiuatad Stream Width 	  B   Estimated Stream  D«pth:   Biffl*

 High H>t«r Mark 	 n    V.loclty  	    Dai  Pr«««nt:   Taa

                           Partly Opan       Partly Shadad
 Canopy Covar:   Opan

 SEDImaT/SUBSTBATB:

 sadiaant  Odora:   Uoraial

 Sodiaiant  Oila:   Abaont
                                                      Industrial



                                                Obvioua Sourcaa

                                               Run 	 n   Pool

                                                  Channalilad:   Taa
                                                                      Oth.c
 Sawaga      Patrolaua      Chaaical

Slight     Hodarata      profuaa
                                          Anaerobic

•: Sludge Savduat Ptpar ribar Sand Belict Shells Other
Are the underaidea of atoi.ea which ara not deeply ambidded black? Yes Bo
Substrata Typa
B drock
B ulder
C bble
0 aval
S nd
Silt
Clay
Percent
Compoait ion
Diameter in Sampling Area
>256-mm (10 in.)
<4-}S6-mm (2.5-10 in.)
2-C4-mm (0.1-2.5 in. )
0.06-2.00-mm (gritty)
.004-.06-mm
<.004-mm (slick)
Percent
Composition
Substrate Type characteristic in Sampling Araa
Datritua Sticks, Wood,
Coarse Plant
Materials (CPOM)
Muck-Hud alack. Very fine
Organic (rPOH)
Marl dray. Shell
pragmenta

HATEB QUALITY

C Dlaaolvad Oiygen pH Conductivity Other

Straaa Typa:  Colduatar       Waraiwatar

Watar Odora:  Boraal      Sawaga      Patrolaun     Chaaical      Bona      Othar

Hatar Surtaca Ofli:   slick      Shaan      Globa      rlacka      Bona

Turbidity:  Claar      Slightly Turbid      Turbid      Opag.ua      Uatar Color  	
HEATBEB COBDITIOHS
PHOTOOBAPH HUMBEB
OBSERVATIONS AHD/OR SKETCB
     Figure 5.1-1.  Physical Characterization/Water Quality Field Data Sheet for use with all Rapid Bioassessment Protocols.

-------
which are associated with sediment in the area of the
sampling station.

Sediment Oils:  Note the term which best describes
the relative amount of any sediment oils observed in
the sampling area.

Sediment Deposits:  Note those deposits described (or
include any other deposits not listed) which are pres-
ent in the sampling area. Also indicate whether the
undersides of rocks not  deeply embedded are black
(which generally indicates low dissolved oxygen or
anaerobic conditions).

Inorganic Substrate Components:  Visually estimate
the relative proportion of each of the seven sub-
strate/particle types  listed that are present in the sam-
pling area.

Organic Substrate  Components: Indicate relative
abundance of each of the three substrate types listed.

             5.1.2 Water Quality

    Information  requested in this section (Figure 5.1-1)
is standard to many aquatic studies and allows for
some comparison  between sites. Additionally,  condi-
tions that may significantly affect aquatic biota are
documented. Documentation  of recent and current
weather conditions is important because of the poten-
tial impact that  weather may have on water quality. To
complete  this phase of the bioassessment, a photo-
graph may be helpful in 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.

Temperature (C), Dissolved Oxygen,  pH, Conduc-
tivity: Measure and record values for  each of the
water quality parameters indicated, using the  appropri-
ate calibrated water quality instrument(s). Note the
type of instrument and  unit number used.

Stream Type: Note the appropriate stream designa-
tion according to State  water quality standards.

Water Odors: Note those odors described (or include
any other odors not listed) that are associated with  the
water in the  sampling area.

Water Surface  Oils: Note the term that best  describes
the relative amount of any oils present on  the water
surface.

Turbidity: Note the term which, based upon  visual
observation, best describes the amount of material
suspended in the water column.
     5.2 HABITAT  ASSESSMENT

   The habitat assessment  matrix (Figure 5.2-1) is
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). 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  sta-
tion  studied. The ratings are then totaled and com-
pared 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.
   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.  How-
ever, 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 Chapter 8.
   Habitat parameters pertinent to the assessment of
habitat quality are separated into three principal cate-
gories: primary, secondary, and tertiary parameters.
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, and estimation of
the flow or velocity and depth regime, have  the widest
score  range (0-20) to reflect their contribution  to hab-
itat quality. The secondary parameters measure the
"macroscale" habitat such as channel morphology
characteristics.  These parameters evaluate: channel
alteration, bottom scouring and deposition, and stream
sinuosity. The secondary parameters  have a  score
range  of 0-15. Tertiary parameters evaluate riparian
and bank structure and comprise three parameters:
bank stability, bank vegetation, and streamside  cover.
These tertiary parameters  include those that  are most
often  ignored in biosurveys.  The tertiary  parameters
have a score range of 0-10.
                                                     5-3

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                                                                              Condition
                  Condition/Parameter
  PRIMARY—SUBSTRATE AND INSTREAM COVER
  1.  Bottom substrate and available cover
  2.  Embeddedness
  3.  Flow/velocity

  SECONDARY-CHANNEL MORPHOLOGY
  4.  Channel alteration
  5.  Bottom scouring and deposition
  6.  Pool/riffle, run/bend ratio

  TERTIARY—RIPARIAN AND BANK STRUCTURE
  7.  Bank stability
  8.  Bank vegetation
  9.  Streamside cover
        Excellent     Good


          16-20       11-15
          16-20       11-15
          16-20       11-15
          12-15
          12-15
          12-15


          9-10
          9-10
          9-10
8-11
8-11
8-11


6-8
6-8
6-8
Fair


6-10
6-10
6-10


4-7
4-7
4-7


3-5
3-5
3-5
Poor


 0-5
 0-5
 0-5


 0-3
 0-3
 0-3


 0-2
 0-2
 0-2
   Habitat evaluations are first made on instream hab-
itat,  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 greatest impact on the
community being studied. The actual habitat assess-
ment process involves rating the  nine parameters as
excellent, good, fair, or poor based on the criteria
included on the Habitat Assessment Field Data Sheet
(Figure  5.2-1).
   A total 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. The station is then classified
on the basis of its similarity to expected conditions (as
represented by the control or reference station), and
its apparent potential to support an acceptable level of
biological health.
   Use of a percent comparability evaluation allows
for regional and stream-size differences which affect
flow or velocity, substrate, and channel morphology.
Some regions are characterized by streams having a
low  channel gradient. Such streams are typically shal-
lower, have a  greater pool/riffle or run/bend ratio, and
less  stable substrate than streams with a steep channel
  Assessment Category

Comparable to Reference
Supporting
Partially Supporting
Non-Supporting
                Percent of
              Comparability

                  >90%
                  75-88%
                  60-73%
                  <58%
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. Using the approach presented here,
these streams may be evaluated relative to other low
gradient streams.
   Listed below is a general explanation  for each of
the nine habitat parameters to be evaluated.

      5.2.1  Primary Parameters—
     Substrate and Instream  Cover

   The  primary instream habitat  characteristics
directly pertinent to the support of aquatic communi-
ties consists of substrate type and stability, availability
of refugia, and migration/passage potential. These pri-
mary habitat parameters are weighted the highest to
reflect their degree of importance to biological
communities.

I.  Bottom Substrate—This refers to the  availability
   of habitat for support of aquatic organisms. A vari-
   ety of substrate materials and habitat types  is
                                                   5-4

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HABITAT
ASSESSMENT FIELD DATA SHEET
Ca t ego ry

1. 'Bottom substrate/
aval 1 abl« cover
2. Embeddedness(b)
3 . 50.15 ens (Scf s ) •
•Floy, at rep. low
flow14'
or
>0 .15 cms ( Scf s I «
Velocity/depth
4. • Channel alteration
S. Bottom scouring and
depos it ion

(a) From Ball 1982.
(b) From Platts et al. 1983.

Excel lent
Greater than 50% rubble,
gravel, submerged logs,
undercut banks, or
other stable habitat.
16-20
Gravel, cobble, and
between 0 and 25 %
surrounded by fine
sediment
16-20
Cold >O.OS cms (2 cfs)
Warm >0.15 cms (5 cfs)
10-20
Slow (<0.3 m/s 1 , deep
(>O.S m); slow, shallow
( <0 .5 m) ; Cast
shallow habitats all
present .
16-20

point bars, and/or
no channelization.
12-15
Less than 51 of the
bottom affected by
scouring and
depos it ion .
12-15


Good
30-50% rubble, gravel
or other stable habitat.
Adequate habitat.
11-15
Gravel , cobble , and
between 25 and 50 t
surrounded by fine
sedi nent
11-15
0.03-0.05 cms (1-2 cfs)
0.05-0.15 cms (2-5 cfs)
11-15
Only 3 of the 4 habitat
categories present
(missing riffles or runs
missing pool s ) .
.11-15

coarse gravel; and/ or
some channelization
pr esent .
8-11
5-30% affected. Scour
at constrictions and
where grades steepen.
8-11


Fair
10-30% rubble, gravel
or other stable habitat.
Habitat availability
less than desirable.
6-10
Gravel, cobble, and
between 50 and 75 t
surrounded by fine
sediment
6-10
0.01-0.03 cms ( .5-1 cfsl
0.03-0.05 cms (1-2 cfs)
6-10
Only 2 of the 4 habitat
categories present
(missing riffles/runs
6-10

on old and new bars;
pools partially filled
ments .on both banks.
. 4-7
30-50% affected.
Deposits and scour at


4-7

Poo r
Less than 10% rubble
gravel or other stable
habitat. Lack of
habitat is obvious.
0-5
Gravel , cobble, and
over 75 % surrounded
by fine sediaent
0-5
(0.01 cms I . S cfs )
<0.03 cms (1 cfs)
0-5
Dominated by on'e
velocity/depth
category (usually
0-5

development; most pools
filled w/silt; and/or
0-3
More than 50% of the
bottom changing
nearly year long.

On ly la rge rocks
in riffle exposed.
0-3

Figure 5.2-1. Habitat Assessment Field Data Sheet for use with all Rapid Bioassessment Protocols.

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HABITAT ASSESSMENT FIELD DATA SHEET (cont.f
Ca t eao r v

6. Pool/riffle, run/bend 5-7. Variety of 7-15. Adequate depth
ratio (distance habitat. Deep riffles in pools and riffles.
between riffles divided and pools. Bends provide habitat.
by s t ream width )
12-15 8-11
7. Bank stability Stable. No evidence Moderately stable.
bank failure. of erosion mostly healed
Side slopes gener- over. Side slopes up to
ally <30% . Little 40% on one bank. Slight
probl em . f loods .
9-10 6-8
8. Bank vegetative Over 80% of the 50-79% of the streambank
covered by vegetation, gravel or
vegetation or boulders larger material.
and cobbl e'.
9-10 6-8
< b ) . . .
t s shrub . is of tree form.
9-10 6-8
Column Totals 	 	
Sco re

Fair
15-25. Occassional
riffle or bend. Bottom
contours provide some
habitat .
4-7
Mode rately unstable.
Mo derate frequency and
size of erosional areas.
Side slopes up to 60%
on some banks. High
erosion potential
during extreme high
flow.
3-5
25-49% of the stream-
by vegetation, gravel,
3-5
Dominant vegetation
is grass or f orbes .
3-5

Poo r
>25. Essentially a
straight stream.
Generally all flat
wa te r or sha 1 low
riffle. Poor
habitat .
0-3
Unstable - Many
eroded areas. Side
slopes > 60% common.
along straight sections
and bends.
0-2
Less than 25% of the
covered by vegetation,
gravel , or larger
ma t e r ia 1 .
0-2
Over 50% of the stream-
bank has no vegetation
and dominant material
is soil, rock, bridge
materials, culverts,
o r nine tail ings .
0-2

Figure 5.2-1.  (Cont.).

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   desirable. The presence of rock and gravel in flow-
   ing 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 habi-
   tat 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, embed-
   dedness is regarded as the stability of cobble sub-
   strate 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) 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 con-
   tour is  relatively flat. Even where a few stations
   may have flows in excess of 0.15 cms,  flow may
   still be the predominating constraint. Therefore, the
   evaluation is based on flow rather than velocity.

   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  relation-
   ship. 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.

      5.2.2 Secondary Parameters—
             Channel Morphology

      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.
4.  Channel Alteration—The character of sediment
   deposits 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 con-
   strictions, 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.
5.  Bottom Scouring and Deposition—These
   parameters relate to the destruction of instream
   habitat resulting  from the problems described
   above. Characteristics to observe are scoured sub-
   strate and degree of siltation in pools and riffles.
   Scouring results  from high velocity flows. The
   potential for scouring is increased by channeliza-
   tion. Deposition and scouring result from the trans-
   port 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).
6.  Pool/Riffle or Run/Bend Ratio—These parameters
   assume that a stream with riffles or bends provides
   more diverse habitat than a straight (run) or uni-
   form 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.

        5.2.3  Tertiary Parameters-
       Riparian and Bank  Structure

      Well-vegetated banks are usually stable regard-
   less 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.
                                                    5-7

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Stream channel alteration downstream of WWTP.
                    5-8

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      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 of the stream cross section  from the nor-
   mal high water line to the lower water line. The
   lower channel defines the  stream width.
7.  Bank Stability—Bank stability is rated by observ-
   ing 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 subject 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, raw areas may not be as sus-
   ceptible to erosion as other soil types.
8.  Bank Vegetative Stability—Bank soil is generally
   held in place by plant root systems. Erosional pro-
   tection may also be provided by boulder, cobble, or
   gravel material.  An estimate of the density of bank
   vegetation (or proportion of boulder, cobble, or
   gravel material)  covering the bank provides  an indi-
   cation of bank stability and potential instream
   sedimentation.
9.  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 bot-
   tom, 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 CPOM source  in allochthonous
   systems.
                                                    5-9

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Poor bank stability with high erosional potential.
    Stream banks stabilized by dense vegetation.
                      5-10

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                     6.  BENTHIC MACROINVERTEBRATE
                       BIOSURVEY  AND DATA  ANALYSIS
   The biosurvey and data analysis components of the
three benthic bioassessment protocols are presented
below. All three protocols have common biosurvey
and data analysis elements. Common elements and
discussions are repeated in each protocol to maintain
discrete protocol integrity.
   Examples of field and  laboratory data sheets
referred to in this chapter are presented for guidance.
The example data sheets do not include headers for
documenting identifier information, and  may be modi-
fied for the needs of different agencies. Descriptive
guidance for use with each data sheet is found in
Appendix A.
   The three protocols consist of three  basic compo-
nents: water quality/physical characteristics (Fig-
ure 5.1-1), habitat assessment (Figure 5.2-1), and a
biosurvey (Figures 6.1-1, 6.2-1, and 6.3-1). The overall
habitat assessment evaluates habitat quality using  the
key environmental parameters described  in Chapter 5.
If a degraded community  is found from  the results of
the biosurvey, habitat information will aid interpreta-
tion of effects relative to the biotic potential of a  site.
The water quality and physical characterizations pro-
vide data on stream  habitat quality as- well as potential
sources and/or causes of impairment.
    6.1 RAPID BIOASSESSMENT
         PROTOCOL  I—Benthic
            Macroinvertebrates
   Rapid Bioassessment Protocol I (RBP I) is a
screening or reconnaissance assessment that involves
systematic -documentation of specific visual observa-
tions made in the field by a trained professional.
RBP I is used to discriminate obviously  impacted and
non-impacted areas from potentially affected areas
requiring further investigation. Use of RBP I allows
rapid screening  of a large number  of sites. Areas
identified for further study can then be rigorously
evaluated using  RBPs II, III, and V; quantitative fish
or benthic surveys; or ambient toxicity studies.
   Because RBP I involves limited data  generation, its
effectiveness depends largely on the experience ("best
professional judgment") of the professional biologist
performing the  assessment. The biologist conducting
RBP I should have professional impact assessment
experience with a knowledge of aquatic ecology and
basic  expertise in benthic macroinvertebrate taxonomy.

            6.1.1 Field  Methods

   The biosurvey component of RBP I focuses on
qualitative sampling of benthic  macroinvertebrates,
supplemented by a preliminary  field examination of
other  aquatic biota (periphyton, macrophytes, slimes,
and fish). Qualitative benthic samples are collected
from  all  available habitats using a dip net or kick net,
or by hand.  Benthic macroinvertebrate orders/families
(e.g.,  families  for Megaloptera  and  Diptera)  collected
are listed on the Biosurvey Field Data Sheet (Fig-
ure 6.1-1), with an estimate of their relative abundance
in the sampling area. Each State agency should
develop its own definitions for abundance categories.
Lower levels of identification, if they are easily deter-
mined, can enhance the  assessment. Any observations
on the relative abundance of other aquatic biota are
also noted; these  observations provide additional infor-
mation on the  presence or absence  of impact.

    6.1.2  Data Analysis Techniques
   Impairment may be indicated by the absence of
generally pollution-sensitive benthic macroinvertebrate
taxa such as Ephemeroptera,  Plecoptera, and Trichop-
tera (EPT);  dominance of generally  pollution-tolerant
groups such  as Oligochaeta or Chironomidae; or over-
all low benthic abundance or taxa richness. Benthic
abundance or taxa richness indicative of impairment is
variable and must be evaluated  with respect to the
waterbody being  evaluated. Some headwater  streams
are naturally unproductive and will  be characterized
by low benthic abundance.and taxa  richness  in their
pristine state. Impairment may also  be indicated by an
overabundance of slimes or filamentous algae in the
area or an absence of expected  fish populations.
   On the basis of the observations made on habitat,
water  quality, physical characteristics, and the qualita-
tive biosurvey, the investigator determines whether
impairment is detected. The determination of impair-
ment  requires the judgment of an experienced profes-
sional. If impairment is  detected, the investigator
provides  an estimation of the probable cause and
source on the Impairment Assessment Sheet  (Fig-
ure 6.1-2). The aquatic biota that indicated an impair-
                                                   6-1

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                              Rapid Bioassessment Protocol I

                                    Biosurvey Field Data Sheet
RELATIVE ABUNDANCE OF AQUATIC BIOTA
Periphyton         01234
Filamentous Algae   01234
Macrophytes        01234


0 = Absent/Not Observed          1 = Rare
                                                      Slimes
                                                      Macroinvertebrates
                                                      Fish
0     1
0     1
0     1
                                              2 = Common
                                                                 3 = Abundant
234
234
234


  4 = Dominant
MACROBENTHOS QUALITATIVE SAMPLE LISTflndrcate Relative Abundance R = Rare, C = Common, A = Abundant, O = Dominant)
Porifera
Hydrozoa
Platyhelminthes
Turbellaria
Hirudinea
Oligochaeta
Isopoda
Amphipoda
pecapoda
Gastropoda
Bivalvia

Anisoptera
Zygoptera
Hemiptera
Coleoptera
Lepidoptera
Sialidae
Corydalidae
Tipulidae
Empididae
Simuliidae
Tabanidae
Culicidae
Chironomidae
Plecoptera
Ephemeroptera
Trichoptera
Other







Rare < 3
Observations
                      Common 3-9
                                                 Abundant > 10
                                                                            Dominant > 50 (Estimate)
        Figure 6.1-1. Biosurvey Field Data Sheet for use with Rapid Bioassessment Protocol I.
                                              6-2

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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:

                 Benthic  macroinvertebrates          Other aquatic communities

                 	 absence  of  EFT  taxa            	  Periphyton

                 	 dominance of  tolerant groups      	 filamentous

                 	 low benthic abundance             	 other

                 	 lov taxa richness              	  Macrophytes

                 	 other                          	  Slimes

                                                           Fish
              3.   Brief  description  of  problem:
                  Year  and  date  of  previous  surveys:
                  Survey  date,  available  in:  	
                  Cause:  (indicate major  cause)    organic enrichment   toxicants   flow

                      habitat  limitations   other 	
                                                      2
                  Estimated  areal extent  of  problem (m )  and length of stream reach

                  affected (m),  where applicable:  	
              6.   Suspected  source(s)  of  problem:

                  	  point  source discharge (name,  type of facility, location)
                  	  construction site runoff
                  	  combined sewer outfall
                  	  silviculture runoff
                  	  animal feedlot
                  	  agricultural runoff
                  	  urban  runoff
                  	  ground water
                  	  other
                        unknown
              Briefly explain:
Figure 6.1-2. Impairment Assessment Sheet for use with macroinvertebrate Rapid Bioassessment Protocols.
                                              6-3

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ment are noted, as are potential sources of pollutants.
The downstream extent of impact is estimated, and
multiplied  by either the approximate stream width at
the estimated fully mixed zone or the  width of the
discharge plume. This calculation provides an estimate
of the area impacted at the site.
    6.2  RAPID BIOASSESSMENT
         PROTOCOL II—Benthic
             Macroinvertebrates
   Rapid Bioassessment Protocol II (RBP II) utilizes
the systematic field collection and analysis of major
benthic taxa. RBP II provides a more intense assess-
ment than RBP I and can detect sites of intermediate
impairment  with relatively little additional  time and
effort.  The protocol can be used to prioritize sites for
more intensive evaluation  (i.e., RBP III, replicate
sampling, ambient toxicity testing, chemical charac-
terization) or can be used in  lieu of RBP I as a
screening technique. RBP II  is based  on RBP III at a
reduced level of effort. RBP  II incorporates the con-
cept of benthic analysis at the family  taxonomic  level,
as advocated by  some States  (e.g., Virginia, Illinois),
and utilizes  field sorting and identification. This level
of effort involves minimal taxonomic  identification and
is sufficient to address the objectives  of RBP II. Furse
et al.  (1984) stated that  family-level classifications are
valuable in developing local site inventories of organ-
isms and in the evaluation of pollution monitoring
programs. The strength of RBP II is a result of sys-
tematic data collection procedures and the  use of
recently developed data analysis techniques.

            6.2.1  Field  Methods

   The biosurvey component of RBP II focuses  on
standardized sampling of benthic macroinvertebrates,
supplemented by a cursory field observation of other
aquatic biota (periphyton, macrophytes, slimes, and
fish) (Figure 6.2-1). Although RBP II emphasizes the
benthic community, the observation of effects  on  other
aquatic biota will support the final evaluation. (This
approach is adapted from Michigan DNR's protocol.)

6.2.1.1 Sample  Collection
   The collection procedure  provides representative
samples of the macroinvertebrate fauna from compara-
ble habitat types at all stations constituting a site
evaluation, and is supplemented with  separate Coarse
Paniculate Organic Matter (CPOM) samples. RBP II
focuses on the riffle/run habitat because it is the most
productive habitat available in stream systems and
includes  many pollution-sensitive taxa of the Scraper
and Filtering Collector Functional Feeding Groups.
The CPOM  sample provides  a measure of effects
(particularly toxicity effects), on a third trophic com-
ponent of the benthic community, the Shredders.
   In sampling situations where a riffle/run habitat
with a rock  substrate is not available, any submerged
fixed  structure will  provide a substrate  for the Scraper
and Filtering Collector Functional Groups emphasized
here.  This allows for the same approach to be used  in
non-wadable streams and large rivers and wadable
streams and  rivers with unstable substrates.

Riffle/Run Sample

   Riffle areas with relatively fast currents and cobble
and gravel substrates generally provide  the most
diverse community. Riffles should be sampled using a
kick net  to collect from an approximately 1 m2 area.
Two 1 m2 riffle samples should  be collected at each
station:  one  from an area of fast current velocity and
one from an area of slower current velocity.  The two
samples  are  composited for processing.  In streams
lacking riffles, run  areas with cobble or gravel  sub-
strate are also appropriate for kick net  sampling.
   Where riffle/run communities with  a rock substrate
are not available, other submerged fixed structures
(e.g. submerged boulders, logs, bridge  abutments, pier
pilings) should be sampled by hand picking.  These
structures provide suitable habitat for the Scrapers and
Filtering Collectors and will allow use  of RBP II for a
wider range of regions and stream orders. Benke et
al. (1984) determined that although submerged wood
substrates, or snags, accounted for only a small por-
tion of the available substrate in  a blackwater river in
Georgia, this habitat provided the greatest taxa rich-
ness and more than half of all benthic  biomass.

CPOM Sample

   In addition to the riffle/run sample  collected for
evaluation of the Scraper and Filtering  Collector
Functional Feeding  Groups, a CPOM sample should
also be collected to provide data on the abundance of
Shredders at the site. Large paniculate  Shredders are
important in forested areas of stream ecosystems rang-
ing from stream orders  1 through 4 (Minshall et al.
1985). The absence of Shredders of large paniculate
material  is characteristic of unstable, poorly retentive
headwater streams in disturbed watersheds or in dry
areas  where  leaf material processing is accomplished
by terrestrial detritivores (Minshall et al. 1985).
McArthur et al.  (1988) reported that very few Shred-
ders were found in  summer leaf packs  in South Caro-
lina because processing was so rapid.
                                                    6-4

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Kick net sampling in riffle area.
              6-5

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                              Rapid Bioassessment Protocol II

                                     Biosurvey Field Data Sheet
RELATIVE ABUNDANCE OF AQUATIC BIOTA
Periphyton
Filamentous
Macrophytes
0 1
Algae 0 1
0 1
0 = Absent/Not Observed
2
2
2
1
3
3
3
= Rare
4 Slimes
4 Macroinvertebrates
4 Fish
0 1
0 1
0 1
2
2
2
2 = Common 3 = Abundant
3
3
3
4
4
4
4 = Dominant
MACROBENTHOS QUALITATIVE SAMPLE LIST
                                                                     List Families Present/Indicate Abundance
Oligochaeta


Gastropoda


Bivalvia


Ephemeroptera




Anisoptera


Zygoptera

Plecoptera


Trichoptera




Coleoptera


Diptera





Other
RIFFLE SAMPLE
FUNCTIONAL FEEDING GROUPS
(Indicate No. ol Individuals Representing Group)
Scrapers
     Filtering Collectors
CPOM SAMPLE  FUNCTIONAL FEEDING GROUPS (Indicate No. ol Individuals Representing Group)
Shredders
                                                   Total Org. in Sample
Observations
       Figure 6.2-1. Biosurvey Field Data Sheet for use with Rapid Bioassessment Protocol II.
                                                6-6

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  . The CPOM sample is processed separately from
the riffle/run sample and used only for characterizing
the Functional Feeding Group representation. Sam-
pling the CPOM component requires a composite col-
lection of various plant parts  such  as leaves, needles,
twigs, bark, or their fragments. Potential sample
sources include leaf packs, shorezones, and other
depositional areas where CPOM may accumulate.
Only the upper surface of litter accumulation in
depositional areas should be sampled to ensure that
they are from the aerobic zone. For the Shredder
community analysis, several handfuls of material
should be adequate. A variety of CPOM forms should
be collected if available. CPOM collected may be
washed in a dip net or a sieve bucket.
   Shredder abundance  is maximum when the CPOM
is about 50 percent decomposed (Cummins et al.
1989).  Care must be taken  to avoid collecting recent
or fully decomposed leaf litter to optimize collection
of the Shredder community. For this CPOM collection
technique, seasonality may  have an important •
influence on Shredder abundance data. For instance,
fast-processing litter (e.g., basswood, alder, maples,
birch) would have the highest Shredder  representation
in the winter (Cummins et al. 1989). The slow-
processing litter (e.g., oaks, rhododendrons, beech,
conifers) would have the highest Shredder representa-
tion in the summer.

6.2.1.2  Sample  Sorting and Identification  .
Riffle/Run Sample
   Sorting and enumeration in the field to obtain a
100-count organism subsample is recommended for
the riffle/run sample. After processing in the field,
the organisms and sample residue  should be preserved
for archiving. Thus, a re-analysis (quality control) or
more thorough processing (e.g., larger counts, more
detailed taxonomy)  would be  possible. The subsam-
pling method described  in this protocol  is based on
Hilsenhoff s Improved Biotic  Index (Hilsenhoff 1987b)
and  is  similar to that used by New York DEC (Bode
1988).  This subsampling technique provides for a con-
sistent unit of effort and a representative estimate of
the benthic fauna.
   The subsampling procedure consists  of evenly dis-
tributing  the  composite sample into a  gridded pan
with a light colored bottom.  Grids are randomly
selected and all organisms  within those  grids are
removed  until approximately  100 organisms are picked
out. Because this subsampling technique is being
applied to samples with live organisms, narcotization
using club soda or tobacco is recommended.  A more
detailed description of this technique may be found  in
Appendix B.
   An alternative method of subsampling live samples
in the field is to simply sort 100 organisms in a ran-
dom manner. Narcotization to slow the organisms is
less important with this subsampling technique. To
lessen sampling bias,  the investigator should pick
smaller, cryptic organisms, as well as the larger, more
obvious organisms.
   All organisms in the subsample should be classi-
fied according to Functional Feeding Group.  Field
classification is important because many  families com-
prise genera and species representing a variety of
functional groups. Knowing the family-level identifica-
tion of the organisms  will generally be insufficient for
categorization by  Functional Feeding Group. Func-
tional Feeding Group  classification can be done in the
field,  on the  basis of  morphological and  behavioral
features, using Cummins and Wilzbach (1985).  Care
should be taken in noting early instars, which may
constitute different Functional  Feeding Groups from
the later instars.
   The  Scraper and Filtering Collector Functional
Groups  are the most important indicators in the riffle/
run community. Numbers of individuals  representing
each of these two groups are recorded on the Biosur-
vey Field Data Sheet  (Figure 6.2-1). All organisms in
the subsample should  be identified to family or order,
enumerated, and recorded, along  with any observa-
tions on abundance of other aquatic biota, on the
Biosurvey Field Data  Sheet. A summary of all benthic
data to be used in the final analysis will  be recorded
on the Data Summary Sheet (Figure 6.2-2) upon
return to the laboratory.
   The use of family-level identification  in this pro-
tocol  is based on Hilsenhoff s  Family .Biotic Index
which uses higher taxonomic levels of identification
(Hilsenhoff 1988). Tolerance characterizations for the
Family  Biotic Index (FBI) and excerpts from Hilsen-
hoff s paper describing the index  are included in
Appendix C. Assessment based on family-level iden-
tifications has been used successfully by  the States of
Virginia and  Illinois.

CPOM Sample
Organisms collected in the supplemental  CPOM sam-
ple are  classified as Shredders or Non-Shredders. Tax-
onomic identification  is not necessary for this
component. The composited CPOM sample may be
field sorted in a  small pan with a light colored bottom
or in the net  or sieve  through  which it was rinsed. (If
a large  number of benthic macroinvertebrates have
been collected, a representative subsampling of 20-60
organisms may be removed for Functional Feeding
Group classification.)  Numbers of individuals
representing the  Shredder Functional Group,  as well
                                                    6-7

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                                         DATA SUMMARY SHEET
Station No.
Station Location
Taxa Richness
FBI (modified)
Functional Feeding Groups
Riffle Community
Scrapers/Filt. Collect.
CPOM Community
Shredders/Total
EPT/Chironomidae
% Contribution (dom. family)
EPT Index
Community Similarity Index
Comments:

















































































Figure 6.2-2. Data Summary Sheet suggested for use in recording benthic data utilized in Rapid Bioassessment Protocol

-------
Field sorting of benthic macroirwertebrate
samples for  Rapid Bioassessment Protocol II.
                   6-9

-------
as total number of macroinvertebrates collected in this
sample, should be recorded on the Biosurvey Field
Data Sheet (Figure 6.2-1) for later analysis.  The
Shredder/Non-Shredder metric may be deemed
optional in rivers or  in some regions where Shredder
abundance is naturally low.  However, the potential
utility of such a  metric for assessing toxicant effects
warrants serious  consideration in this bioassessment
approach.


    6.2.2  Data  Analysis Techniques

   Biological impairment of the benthic community
may be indicated by  the absence of generally
pollution-sensitive macroinvertebrate taxa  such as
Ephemeroptera, Plecoptera,  and Trichoptera (EFT);
excess dominance by any particular taxon, especially
pollution-tolerant forms such as some Chironomidae
and Oligochaeta taxa; low overall taxa richness; or
appreciable shifts in  community composition relative
to the reference condition. Impairment may  also be
indicated by an overabundance  of fungal  slimes or
filamentous algae, or an absence of expected popula-
tions of fish.  All of these indicators can be evaluated
using the sampling data generated  in RBP II.
   On the  basis of observations made in the assess-
ment of habitat, water quality, physical characteristics,
and the qualitative biosurvey, the investigator con-
cludes whether impairment is detected. If impairment
is detected, an estimation of the probable cause and
source is provided on the Impairment Assessment
Sheet (Figure 6.1-2).  The aquatic biota that indicated
an impairment are noted along with observed indica-
tions of potential problem sources. The downstream
extent of impact  is estimated and multiplied by
appropriate stream width to provide an estimate of the
areal extent of the  problem.
   The data analysis  scheme used in RBP II integrates
several community, population, and functional
parameters  into a single evaluation of biotic integrity
(Table 6.2-1). Each parameter, or metric,  measures  a
different component  of community structure and has a
different range of sensitivity to pollution  stress (Fig-
ure 8.2-1).  This integrated approach provides more
assurance of a valid  assessment because a variety of
parameters  are evaluated. Deficiency of any one met-
ric in a particular situation should not  invalidate the
entjre approach.
   The eight metrics used in RBP II are  the same as
those in RBP III, but the scoring criteria used to
evaluate the metrics  have been  modified to accommo-
date the less rigorous taxonomy (family-level identifi-
cations) of RBP  II. The integrated data analysis
(Figure 6.2-3) is performed  as  follows. Using the raw
benthic data, a numerical value is calculated for each
metric. Calculated values are then compared to values
derived from either a reference site within the same
region, a reference database applicable to the region,
or a suitable control station on the same stream.  Each
metric is then assigned a score according to the com-
parability (percent similarity) of calculated and refer-
ence values. Scores for the eight metrics are then
totaled and compared to the total metric  score for the
reference station. The percent comparison between the
total scores provides a final evaluation of biological
condition.
   The criteria to be used  for scoring the eight met-
rics were derived from an evaluation  of pilot study
results (Section 6.4); certain compliance  monitoring
requirements now in use (Vermont Department of
Environmental Conservation 1987); and discussions
with various aquatic biologists regarding the level of
detection considered dependable for certain metrics.
However, these criteria may need to be adjusted for
use in particular regions.
   Inherent  variability in each metric was considered
in establishing percent comparability  criteria. The
metrics based  on taxa richness, FBI,  and EFT Indices
have low variability (Resh  1988). This variability is
accounted for in the criteria for characterization of
biological condition (Figure 6.2-3), based on existing
data.  For metrics based on standard taxa richness and
FBI and EPT Indices, differences  of 10-20 percent
relative to the reference condition would be considered
nominal, and the station being assessed would receive
the maximum  metric score. Because increasing FBI
values denote worsening biological condition, percent
difference for this metric is calculated by dividing the
reference value by the  value for the station of
comparison.
   Metrics that utilize ratios fluctuate more widely,
however, and comparing percent differences between
ratios (ratios of ratios) will compound the variability.
Scoring increments are therefore  set at broad intervals
of 25 percent or greater. For metrics  based on  Func-
tional Feeding Group ratios, Cummins (1987, personal
communication) contends that differences as great as
50 percent from the  reference may be acceptable, but
differences in the range of 50-100 percent are not
only important but discriminate degrees of impact
more clearly.
   The contribution  of the dominant  taxon to total
abundance is a simple estimator of evenness. Scoring
criteria are based on theoretical considerations  rather
than direct comparison with a reference.
   The Community Loss Index (a representative
similarity index) already incorporates a comparison
with a reference. Therefore, actual index values are
used in scoring.
   The metrics used to evaluate the benthic data  and
their significance are explained below.
                                                     6-10

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TABLE 6.2-1 CRITERIA*3* FOR CHARACTERIZATION



1.


2.

3.


4.

5.



6.



7.

8.

Metric

Taxa Richness


Family Biotic Index (modified)

Ratio of Scrapers/Filtering Collectors*


Ratio of EPT and Chironomid Abundances

% Contribution of Dominant Family



EPT Index


/c\
Community Similarity Indexv

Ratio of Shredders/Total* '
OF BIOLOGICAL CONDITION FOR RAPID BIOASSESSMENT PROTOCOL II

Non-Impaired
hn tn «-t IB O
o n n x o
i-l n O T3 3
C T> IB T3
VI O 3" O 0»
i-i c n IB 01
IB n a. cr
a IB in i->
3 n C IB
x-x n >-*•
W O C r^ r»
M. 0 0 3" O
N 3 <-> r—
IB "O C 3 «-»
on 3-
3 M. • 3
O-'rr CT
3" O O O W
0) 3 -a o <-•
cr rr n
H- 01 H- IB M
~ 3 3 W M.
01 0. C t-. ,-r
n 3 O C
a. 3 01
XI O O • rr
C 3 O M.
01 M. 3 0
p-> 3 3 CD 3
M. 01 C 01
r-r 3 3 1— r>
•< n M. o> o
• IB ~ 3
s^>< o cr
IB IB
0-
Biological Condition
Moderately Impaired
01 -«1
3 IB
ID
O
n ~
3 Oi
in x
• 01
a.
IB IB
a.
C rf
o o
o' o
3 in
in
3 0
tTt
PI

H O
in

3
CL t->*
(B 3
X r»
• O
t— *
IB
I-I
1

Severely Impaired
•-» O 1J
01 H IB
x oq «
U 01

w" x
O 3 01
3 M
>-"• 13
>< r,
•-• IB
rr T W
O .il IB
t— 3 3
IB ^
n Q. •
0) O
3 3
3 1-h
O 01
n -r 3-
W IB M-

3 3"

M -< Q.
3 IB
MOD
3 M
•O (B M.
IB 0 £
M 1 IB
IB M
3 »-t
rr < 0
• O rt%
(a)  Scoring criteria are generally based on percent comparability to the reference sta±ion...
(b)  Determination of Functional Feeding Group is independent of taxonomic grouping.
(c)  Community Similarity Indices are used in comparison to a reference station.

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                                                Site-Specific Study
                                               Sampling &  Analysis
          CRITERIA FOR CHARACTERIZATION OF BIOLOGICAL CONDITION FOR PROTOCOL II


1.
2.
3.
4.
5.
6.
7.
8.

Metric
Taxa Richness'3'
Family Biotic Index (modified)(b)
Ratio of Scrapers/Filt. Collectors'2-0'
Ratio of EPT and Chironomid Abundances'3'
% Contribution of Dominant Family'2'
EPT Index'8'
Community Loss Index'e'
Ratio of Shredders/Total'a-c)

6
>80%
>85%
>50%
>75%
<30%
>90%
<0.5
>50%
Biological Condition Scoring Criteria
3
40-80%
50-85%
25-50%
25-75%
30-50%
70-90%
0.5-4.0
25-50%

0
<40%
<50%
<25%
<25%
>50%
<70%
>4.0
<25%
(a)  Score is a ratio of study site to reference site x 100.
(b)  Score is a ratio of reference site to study site x 100.
(c)  Determination of Functional Feeding Group is independent of taxonomic grouping.
(d)  Scoring criteria evaluate actual percent contribution, not percent comparability to the reference station.
(e)  Range of values obtained. A comparison to the reference station is incorporated in these indices.
                                               BIOASSESSMENT
                     % Comp.
                      to  Ref.
                      Score'"'

                      >79%     Non-impaired
 Biological Condition
	Category	
Attributes
                      29-72%    Moderately impaired
                       <21%     Severely impaired.
                        Comparable to the best situation
                        to be expected within an ecoregion.
                        Balanced trophic structure. Optimum
                        community structure (composition and
                        dominance)  for stream size and habi-
                        tat quality.
                        Fewer species due to loss of most
                        intolerant forms. Reduction  in EPT
                        index.
                        Few species present. If high den-
                        sities of organisms, then dominated
                        by one or two taxa. Only tolerant
                        organisms present.
                     (a) Percentage values obtained that are intermediate to the above ranges
                        will require subjective judgement as to the correct placement. Use
                        of the habitat assessment and physicochemical data may be necessary to aid in
                        the decision process.
                                                Recommendations
     Figure 6.2-3.  Flowchart of bioassessment approach advocated for Rapid Bioassessment Protocol
                                                      6-12

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Riffle/Run Sample

Metric 1.  Taxa Richness

             Reflects health of the community
          through a measurement of the variety of
          taxa (total number of families) present.
          Generally increases with increasing water
          quality, habitat diversity, and habitat suit-
          ability. Sampling of highly similar habitats
          will reduce the variability in this metric
          attributable to factors such as current speed
          and substrate type. Some pristine headwater
          streams may be naturally unproductive, sup-
          porting only a very limited number of taxa.
          In these situations, organic enrichment may
          result in an increased number of taxa
          (including EPT taxa).

Metric 2.  Modified Family Biotic Index

             Tolerance values range from 0 to 10 for
          families and increase as water quality
          decreases. The index was developed by Hil-
          senhoff (Hilsenhoff 1988) to summarize the
          various tolerances of the benthic arthropod
          community with a single value. The Modi-
          fied Family Biotic Index was developed to
          detect organic pollution and is based on the
          original species-level index (Hilsenhoff
          1982).  Tolerance values for each family
          were developed by weighting' species
          according to their relative abundance in the
          State of Wisconsin.
             The family-level index has been modi-
          fied for this document  to include organisms
          other than just arthropods using the genus
          and species-level biotic index developed by
          the State of New York  (Bode 1988).  The
          formula for calculating the Family Biotic
          Index is:
                                 X: tj
                                 -
          where

          Xj = number of individuals within a taxon

          tj = tolerance value of a taxon

          n = total number of organisms  in the sample

              Hilsenhoffs family-level tolerance values
          may require modification for some regions.
          Alternative tolerance  classifications and
          biotic indices have been developed by some
          State agencies (Appendix C). Additional
          biotic indices are listed in U.S. EPA (1983).
             Although the FBI may be applicable for
          toxic pollutants, it has only been evaluated
          for organic pollutants. The State of Wiscon-
          sin is conducting a study to evaluate the
          ability of Hilsenhoffs index to detect non-
          organic effects.                         '.

Metric 3.  Ratio of Scraper and Filtering Collector
          Functional  Feeding Groups

             The Scraper and Filtering Collector met-
          ric reflects the  riffle/run community food-
          base.  When compared to a reference site,
          shifts in the dominance of a particular feed-
          ing type indicate a community responding
          to an  overabundance of a particular food
          source. The predominant feeding strategy
          reflects the type of impact detected.  Assign-
          ment  of individuals to Functional Feeding
          Groups is independent of taxonomy, with
          some families representing several func-
          tional groups.
             A description of the Functional Feeding
          Group concept can be found in Cummins
          (1973) and Merritt and Cummins (1984).
          Functional  Feeding Group designations for
          most aquatic insect families may be  found
          in Merritt and  Cummins (1984). Most
          aquatic insects  can also be classified to
          Functional  Feeding Group in the field,  on
          the basis of morphological and behavioral
          features, using  Cummins and Wilzbach
          (1985).
             The relative abundance of Scrapers and
          Filtering Collectors in the riffle/run  habitat
          is an  indication of the periphyton commu-
          nity composition, availability of suspended
          Fine Paniculate Organic Material (FPOM),
          and availability of attachment  sites for filter-
          ing. Scrapers increase with increased dia-
          tom abundance and decrease as filamentous
          algae and aquatic mosses  (which scrapers
          cannot efficiently harvest) increase. How-
          ever,  filamentous algae and aquatic mosses
          provide good attachment sites for Filtering
          Collectors, and the organic enrichment
          often  responsible for overabundance of
          filamentous algae can also provide FPOM
          that is utilized  by the Filterers.
             Filtering Collectors are also sensitive to
          toxicants bound to fine particles and should
          be the first group to decrease when  exposed
                                                     6-13

-------
          to steady sources of such bound toxicants.
          This situation is often associated with point-
          source discharges where certain toxicants
          adsorb readily to dissolved organic matter
          (DOM) forming FPOM during flocculation.
          Toxicants thus become available to Filterers
          via FPOM. The Scraper to Filtering Collec-
          tor ratio may not be a good indicator of
          organic enrichment if adsorbing toxicants
          are present. In these instances the FBI and
          EPT Index may provide additional insight.
          Qualitative field observations on periphyton
          abundance may also be helpful in interpret-
          ing results.

Metric 4. Ratio of EPT and Chironomidae
          Abundances

             The EPT and  Chironomidae abundance
          ratio uses relative abundance of these indi-
          cator groups  (Ephemeroptera, Plecoptera,
          Trichoptera, and Chironomidae) as a mea-
          sure of community  balance. Good biotic
          condition is reflected in communities with
          an even distribution,among, all four major
          groups and with substantial representation
          in the sensitive groups Ephemeroptera,
          Plecoptera, and Trichoptera.  Skewed popu-
          lations having a disproportionate number of
          the Chironomidae relative to  the more sen-
          sitive insect groups may indicate environ-
          mental stress (Ferrington  1987,  Shackleford
          1988). Certain species of some genera such
          as Cricotopus are highly tolerant  (Lenat
          1983, Mount  et al.  1984)  and as oppor-
          tunists may become numerically dominant
          in habitats exposed  to metal discharges
          where EPT taxa are not abundant, thereby
          providing a good indicator of toxicant stress
          (Winner et al. 1980).  Clements et al. (1988)
          found that mayflies were more sensitive
          than chironomids to exposure levels  of 15  to
          32 ^g/L of copper.  Chironomids tend to
          become increasingly dominant in terms of
          percent taxonomic composition and relative
          abundance along a  gradient of increasing
          enrichment or heavy metals concentration
          (Ferrington 1987).
             An alternative to the ratio of EPT and
          Chironomidae abundance metric is the Indi-
          cator Assemblage Index (IAI) developed by
          Shackleford (1988). The IAI  integrates the
          relative abundances of the EPT taxonomic
          groups and the relative  abundances of
          chironomids and annelids upstream and
          downstream of a pollutant source to evalu-
          ate impairment. The IAI  may be a valuable
          metric in areas where the annelid commu-
          nity may fluctuate substantially in response
          to pollutant stress.

Metric 5.  Percent Contribution of Dominant Family

             The percent contribution of the domi-
          nant family to the total number of organ-
          isms uses abundance of the numerically
          dominant taxon relative to the rest of the
          population as an  indication of community
          balance at the family-level.  A community
          dominated by relatively few families would
          indicate environmental stress.  (This metric
          may be redundant if the Pinkham and Pear-
          son Similarity Index is used as a commu-
          nity similarity index for metric number 7.)

Metric 6.  EPT Index

             The EPT Index generally increases with
          increasing water  quality. The EPT Index
          value is the total number of distinct taxa
          within the groups Ephemeroptera, Plecop-
          tera, and Trichoptera.  The EPT Index value
          summarizes the taxa richness within the
          insect groups that are  generally considered
          pollution  sensitive. This was developed for
          species-level identifications; however, the
          concept is valid for  use at family-level
          identifications.
             Headwater streams which are naturally
          unproductive  may experience an increase in
          taxa (including EPT taxa) in response to
          organic enrichment.

Metric 7.  Community Similarity Indices

             Community Similarity Indices are used
          in situations where a reference community
          exists, either through sampling or through
          prediction for a  region. Data sources or
          ecological data files may  be available to
          predict a  reference community to be used
          for comparison. The combined information
          provided through a regional analysis and
          EPA's ERAFT ecological database (Dawson
          and Hellenthal 1986) may be useful for this
          analysis. These indices are designed to be
          used with either  species level  identifications
          or higher taxonomic levels. Three of the
          many community similarity indices available
          are discussed below:
                                                     6-14

-------
  Community Loss Index—Measures the
  loss of benthic taxa between a reference
  station and the station of comparison.
  The Community Loss Index was devel-
  oped by Courtemanch and Dav'ies (1987)
  and is an index of compositional dis-
  similarity, with values increasing as the
  degree of dissimilarity with the reference
  station increases. Values range from  0 to
  "infinity." Based on preliminary data
  analysis, this index provides greater dis-
  crimination than either of the following
  two community similarity indices.

  Jaccard Coefficient of Community
  Similarity—Measures the degree of
  similarity in taxonomic composition
  between two stations  in terms of taxon
  presence or absence.  The Jaccard Coeffi-
  cient discriminates between highly similar
  collections.  Coefficient values, ranging
  from 0 to 1.0,  increase as the degree of
  similarity with the reference station
  increases. See Jaccard (1912), Boesch
  (1977), and U.S. EPA (1983) for more
  detail. The formulae for the Community
  Loss Index and the Jaccard Coefficient
  are
          Community Loss =
                             d-a
       Jaccard Coefficient =
                            a + b + c
  where
  a = number of taxa common to both
       samples
  b = number of taxa present in  Sample
       B but not A
  c = number of taxa present in  Sample
       A but not B
  d = total number of taxa present in
       Sample A
  e = total number of taxa present in
       Sample B
  Sample A = reference station (or mean
              of reference database)
  Sample B = station of comparison

• Pinkham and Pearson Community
  Similarity Index—Incorporates abundance
  and compositional information and can
            be calculated with either percentages or
            numbers. A weighting factor can be
            added that assigns more significance to
            dominant taxa. See Pinkham and Pearson
            (1976) and U.S. EPA (1983) for more
            detail. The formula is
         S.I.
                   min (xia,
             ab "  max (xja,
            where
!is  .^fi  /2
X      V.  '
                                   weighting  factor
            xia, xib = number of individuals in the ith
                    taxon in Sample A or B

             Other community similarity indices sug-
          gested by reviewers of this document
          include Spearman's  Rank Correlation
          (Snedecor and Cochran  1967), Morisita's
          Index (Morisita 1959), Biotic Condition
          Index (Winget and Mangum  1979),  and
          Bray-Curtis Index (Bray  and  Curtis 1957,
          Whittaker  1952). Calculation of a chi-
          square "goodness of fit" (Cochran  1952)
          may also be appropriate.

CPOM Sample

Metric 8.  Ratio of Shredder Functional Feeding
          Group and Total Number of Individuals
          Collected

             Also  based on the  Functional Feeding
          Group concept, the abundance of the Shred-
          der Functional Group  relative to  the abun-
          dance of all other Functional Groups allows
          evaluation of potential impairment as indi-
          cated by the CPOM-based Shredder com-
          munity. Shredders are sensitive to riparian
          zone  impacts and are particularly good indi-
          cators of toxic effects when the toxicants
          involved are readily adsorbed to the CPOM
          and either  affect microbial communities
          colonizing the  CPOM or the Shredders
          directly (Cummins 1987, personal
          communication).
             The degree of toxicant effects on Shred-
          ders versus Filterers depends on  the nature
          of the toxicants and the organic particle
          adsorption efficiency. Generally, as  the size
          of the particle  decreases, the adsorption
          efficiency increases as a function of the
          increased surface to volume ratio (Hargrove
          1972). Because water-borne toxicants are
          readily adsorbed to FPOM, toxicants of a
                                         6-15

-------
          terrestrial source (e.g., pesticides, herbi-
          cides) accumulate on CPOM prior to leaf
          fall thus  having a substantial effect on
          Shredders (Swift et al. 1988a and 1988b).
          The focus of this approach is on a compari-
          son to the reference community which
          should have a reasonable representation of
          Shredders as dictated by seasonally, region,
          and climate. This allows for an examination
          of Shredder or Collector "relative"  abun-
          dance as indicators of toxicity.

   The data collected in the  100-organism riffle/run
subsample and the  CPOM sample are summarized
according to  the information  required for each metric
and entered on the Data Summary Sheet (Fig-
ure 6.2-2).
   Each metric result is given a score based on per-
cent comparability  to a reference station. Scores are
totaled and compared to the total metric score for the
reference station. The percent comparison between the
total scores provides a final evaluation of biological
condition.  Values obtained may sometimes be  inter-
mediate to established ranges and require some judg-
ment as  to assessment of biological condition. In these
instances, habitat assessment, physical characteriza-
tion, and water quality data may aid in the evaluation
process.
    6.3  RAPID BIOASSESSMENT
        PROTOCOL III—Benthic
             Macroinvertebrates
   Rapid Bioassessment Protocol in (RBP III) is a
more rigorous bioassessment technique than RBP II,
involving systematic field collection and subsequent
lab analysis in order to allow detection of more subtle
degrees of impairment. Discrimination of four levels
of impairment should be possible with this assess-
ment. Although  Protocol III requires more detailed
taxonomy than can ordinarily be accomplished in the
field, lab analysis procedures emphasize a minimal
level of effort to ensure the protocol's time- and cost-
effectiveness. Where differences among stations are
subtle, however, more detailed sample analyses (e.g.,
enumeration of larger subsamples) or processing of a
greater number of samples (to define replicability or
assess more habitats) may be  necessary to resolve
such differences.
   Data  provided by RBP III can be used lo prioritize
sites for  more intensive evaluation (e.g., quantitative
biological surveys, ambient toxicity testing, chemical
characterization). Besides providing a means of evalu-
ating effects among stations, this protocol provides a
basis for monitoring trends in benthic community
structure that  might be attributable to improvement or
worsening of conditions over time.

            6.3.1  Field  Methods

    The biosurvey component  of RBP III focuses on
the sampling of benthic macroinvertebrates supple-
mented by cursory field observation of the periphyton,
macrophyton, slime, and fish  communities. The infor-
mation on observed effects upon other aquatic biota
is recorded on the Biosurvey  Field Data Sheet (Fig-
ure 6.3-1) and may be used to support or further
evaluate benthic data.
    The habitat assessment evaluates habitat quality on
the basis of key parameters of the waterbody and sur-
rounding land as described in Chapter 5.  Habitat
assessment is  especially important in situations where
benthos and other biological communities indicate an
impairment. In these  instances,  an evaluation of habi-
tat quality will aid in the interpretation of effects  rela-
tive to a site's biotic potential. The water quality/
physical characterization provides  pertinent data on
habitat quality as well as potential sources or causes
, of impairment.

6.3.1.1 Sample Collection
    The purpose of the standardized collection proce-
dure is to obtain representative  samples of the macro-
 invertebrate fauna from comparably productive habitat
types available at all  stations constituting  a site evalua-
tion, supplemented with separate CPOM samples.
This protocol focuses on the riffle/run habitat as the
 most productive habitat available in stream systems.
The riffle/run benthic community includes many
 representatives of the Scraper and Filtering Collector
 Functional Feeding Groups. Riffle/run sampling is
 supplemented with collection  of a CPOM sample. The
 CPOM sample provides  a measure of effects on a
 third trophic component of the benthic community, the
 Shredders.
    Where riffle/run habitat with a rock substrate is
 not available,  other submerged fixed structures e.g.,
 submerged boulders,  logs, bridge  abutments, pier pil-
 ings, will provide a substrate  for the  Scraper and
 Filtering Collector Functional Groups emphasized
here. Sampling submerged fixed structures would also
be appropriate in non-wadable streams and large rivers
and wadabie streams  and rivers with  unstable
substrates.
                                                    6-16

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                             Rapid Bioassessment Protocol III

                                    Biosurvey Field Data Sheet
RELATIVE ABUNDANCE OF AQUATIC BIOTA
Periphyton          0
Filamentous Algae   0
Macrophytes         0


0 = Absent/Not Observed
      3
      3
      3


1 = Rare
Slimes             0     1
Macroinvertebrates   0     1
Fish                0     1
                                               2 = Common
                                                                   3 = Abundant
2     3
2     3
2     3
                                                                                        4 = Dominant
MACROBENTHOS QUALITATIVE SAMPLE LIST'lndicate Relative Abundance R = Rare, C = Common, A = Abundant, D = Dominant)
Porilera
Hydrozoa
Platyhelminthes
Turbellaria
Hirudinea
Oligochaeta
Isopoda
Amphipoda
Decapoda
Gastropoda
Bivalvia

Anisoptera
Zygoptera
Hemiptera
Coleoptera
Lepidoptera
Sialidae
Corydalidae
Tipulidae
Empididae
Simuliidae
Tabanidae
Culicidae
Chironomidae
Plecoptera
Ephemeroptera
Trichoptera
Other







Rare < 3
                       Common 3-9
                                                  Abundant > 10
                                                                              Dominant > 50 (Estimate)
CPOM SAMPLE  FUNCTIONAL FEEDING GROUPS  (Indicate No. ol Individuals Representing Group)
Shredders
                                                  Total Org. in Sample
Observations
       Figure 6.3-1.  Biosurvey Field Data Sheet for use with Rapid Bioassessment Protocol III.
                                               6-17

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Riffle/Run Sample

   In most situations, a riffle area with relatively fast
current and a cobble and gravel substrate provide the
most diverse community. Riffles should be sampled
using a kick net to collect  from an approximately
1 m2 area. Two 1 m2 riffle samples should be col-
lected at each station: one  from an area of fast current
velocity and one  from an area of slower current veloc-
ity. The two samples are composited for processing.
In streams lacking riffles, run areas with cobble or
gravel substrate are also appropriate for kick net
sampling.
   Where a riffle/run community with a rock sub-
strate is not available, other submerged fixed struc-
tures, e.g., submerged boulders, logs, bridge
abutments, pier pilings,  should be sampled by hand
picking. These structures provide suitable habitat for
the Scrapers and  Filtering Collectors and will allow
use of RBP III for a wider range of regions and
stream orders.  Evaluation of benthic production in a
blackwater stream in Georgia by Benke et al. (1984)
indicated that although submerged wood substrates
(snags) comprised a minor portion of available sub-
strate, the greatest taxa richness and more than half of
all benthic biomass were associated  with this habitat.
   Field inspection of the sample is recommended to
obtain a preliminary assessment of presence and rela-
tive abundance of major groups (to be indicated on
the Biosurvey Field'Data Sheet, Figure 6.3-1), and to
determine if the sampling effort was adequate to
obtain at least  100 organisms. In some samples from
severely impaired areas, organism abundance may not
total 100 organisms. The samples collected at the two
current velocities from the  same habitat are com-
posited, preserved,  labeled, and returned to the
laboratory for processing.

CPOM Sample

   In addition  to the riffle/run sample collected  for
evaluation of the  Scraper and Filtering Collector
Functional Feeding Groups, a CPOM sample should
also be collected  to provide data on  the relative abun-
dance of the Shredders at the site. Shredders of large
paniculate material are important in forested areas of
stream ecosystems ranging  from stream orders 1
through 4 (Minshall et al.  1985). The absence of large
particulate Shredders is characteristic of unstable,
poorly retentive headwater  streams in disturbed
watersheds or in  dry areas  where leaf material
processing is accomplished by terrestrial detritivores
(Minshall et al. 1985). McArthur et al. (1988)
reported that very few Shredders were found in sum-
mer leaf packs in South Carolina because processing
was  so rapid.
   The CPOM sample is processed separately from
the riffle/run sample and used for Functional  Feeding
Group characterization. Sampling of the CPOM com-
ponent requires a composite collection of any of a
variety of forms of CPOM  (plant parts  such as leaves,
needles, twigs,  bark, or fragments of these).  Potential
sample sources include leaf packs and shorezone areas
where CPOM may accumulate. For the Shredder com-
munity analysis, collection  of several handfuls of
material should be adequate. A variety  of CPOM
forms should be collected if they are available. Mate-
rial collected may be washed in a dip net or  a sieve
bucket.
   Maximum Shredder abundance is obtained when
the CPOM is about 50 percent decomposed (Cum-
mins et al. 1989). Care must be taken to avoid col-
lecting recent or fully decomposed leaf litter to
optimize collection of the Shredder community.  Sea-
sonality may have an important influence on  Shredder
abundance data. For instance,  fast-processing  litter
(e.g., basswood, alder, maples, birch) would  have
the highest Shredder representation in the winter
(Cummins et al. 1989). The slow-processing  litter
(e.g., oaks, rhododendrons, beech, conifers)  would
have the highest Shredder representation in the summer.

6.3.1.2  Field Processing
of the CPOM Sample

   Organisms collected in the supplemental CPOM
sample are classified as either Shredders or Non-
Shredders. Taxonomic identification is not necessary
for this component. The composited CPOM sample
may be sorted in the field in a small pan with a light
colored bottom. (If a large  number of benthic macro-
invertebrates has been collected, a representative sub-
sampling of 20-60 organisms may be removed for
Functional Feeding  Group classification.) Numbers of
individuals representing the Shredder Functional
Group, as well as total number of macroinvertebrates
collected in  this sample, should be recorded on the
Biosurvey Field Data Sheet (Figure 6.3-1) for  later
analysis.

             6.3.2 Lab Methods
6.3.2.1  Sample Sorting and Identification
   A 100-organism subsample is recommended as a
time-saving sorting procedure for use with the riffle/
run sample.  The subsampling method described for
use in this protocol is based on that used for  Hilsen-
hoff s Biotic Index (Hilsenhoff 1987b) and is similar to
that used by New York DEC (Bode 1988) and in
Arkansas (Shackleford  1988). The subsampling proce-
                                                    6-18

-------
dure consists of evenly distributing the composite
sample in a gridded pan with a light-colored bottom.
As grids are randomly selected, all organisms within
those grids are removed, until at  least 100 organisms
have been selected from  the sample. This method  of
subsampling provides a representative estimate of the
benthic  fauna as well as  a consistent unit of effort. A
more detailed description of this  technique may be
found in Appendix  B. Although pilot study results
(Section  6.4.6) indicated that  a 100-organism subsam-
ple is sufficient, a 200- or 300-organism subsample
may be preferred, depending on  investigator prefer-
ence, budget constraints, and  individual  sample
characteristics. Some agencies may prefer to expend
additional resources to process whole samples instead
of subsampling.
   All benthic macroinvertebrates in the subsample
(or sample) should  be identified  to the lowest posi-
tively identified taxonomic level  (generally genus or
species), enumerated, and recorded on the Laboratory
Bench Sheet (Figure 6.3-2). Based on the taxonomic
identifications, Functional Feeding Group classifica-
tions can be assigned for most aquatic insects using a
reference such  as Merritt and Cummins  (1984). Once
a Functional Feeding Group  classification list has
been established, it  can be incorporated  into the com-
puter analysis for computation of the metrics. Care
should be taken to  note the presence of  early instars
which may  represent different Functional Feeding
Groups from later instars. The Scraper and Filtering
Collector Functional Groups are  considered the
important indicators in the riffle/run community; if
this  metric is not calculated using a computer pro-
gram, numbers of individuals representing each of
these two groups are recorded on the Laboratory
Bench Sheet (Figure 6.3-2).

     6.3.3  Data Analysis Techniques

Based on observations made in assessing habitat,
water quality, physical characteristics, and the qualita-
tive  biosurvey, the  investigator makes a  preliminary
judgment on the presence or absence of biological
impairment and an estimation of probable cause
and  source  on  the Impairment Assessment Sheet
(Figure 6.1-2).
   The integrated benthic data analysis  is performed
as follows. Using the raw benthic data,  a numerical
value is calculated  for each metric. Calculated values
are then compared  to values derived  from either an
unimpaired reference site within  the same region or a
suitable  control station on the same stream. Each met-
ric is then assigned a score according to the compara-
bility (percent  similarity) of calculated and reference
values.  Scores for the eight metrics are  then totaled
and compared to the total metric score for the refer-
ence station. The percent comparison between the
total scores provides a final evaluation of biological
condition.
   Criteria to be used for scoring the eight metrics
were derived from  an evaluation of pilot study results
(Section 6.4), certain project compliance monitoring
requirements now in use (Vermont Department of
Environmental  Conservation 1987), and  discussions
with various aquatic biologists regarding the level of
detection considered dependable for certain metrics.
However, it is envisioned that these criteria may need
to be adjusted for use in particular regions.
   Inherent variability in each metric was considered
in establishing  percent comparability criteria. The
metrics based on taxa richness, HBI,  and EFT indices
have low variability (Resh  1988). This variability  is
accounted  for in the criteria for characterization of
biological  condition (Figure 6.2-3) based on existing
data. For metrics based on standard taxa richness and
HBI and EFT Indices, differences of 10-20 percent
relative to  the reference condition would be considered
nominal, and the station being assessed  would receive
the maximum metric score. Because increasing HBI
values  denote worsening biological condition, percent
difference  for this metric is calculated by dividing the
reference value by the value for the station of
comparison.
   Metrics that utilize ratios will fluctuate more
widely, however, and comparing percent differences
between ratios  (ratios of ratios) will compound the
variability.  Scoring increments are therefore set at
broad intervals  of 25 percent or greater. For metrics
based on Functional Feeding Group ratios, Cummins
(1987, personal  communication) contends that differ-
ences as great as 50 percent from the reference may
be acceptable, but differences in the range of 50-100
percent are not only important but discriminate
degrees of impact more clearly.
   The percent contribution of the dominant taxon to
total abundance is a simple estimator of evenness.
Scoring criteria are based on theoretical considera-
tions rather than direct comparison with a reference.
   The Community Loss Index already  incorporates
comparison with a  reference. Therefore, actual index
values  are  used in scoring.
   Analysis of the benthic data combines several com-
munity population and functional parameters. An
integrated assessment is used, based on  eight metrics
(Table  6.3-1). Each metric has a different range of
sensitivity  measuring a slightly different component of
community structure  (Figure 8.2-1). The data collected
in the  100-organism riffle/run subsample and the
CPOM sample are  summarized according to the  infor-
mation required for each metric and entered on the
                                                    6-19

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                       LABORATORY BENCH SHEET
                                   Number of Organisms
Station Number
Station Location
Species Name



















Total Organisms
Number of Taxa
































































































Figure 6.3-2. Laboratory Bench Sheet suggested for use in recording benthic
             data utilized in Rapid Bioassessment Protocol III.
                                 6-20

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to
       TABLE 6.3-1  CRITERIA
                            (a)
FOR CHARACTERIZATION OF BIOLOGICAL CONDITION FOR RAPID BIOASSESSMENT PROTOCOL III



1.


2.

3.


A.



5.


6.

7.
8.


Metric
Taxa Richness


Hilsenhoff Biotic Index (modified)

Ratio of Scrapers/Filtering Collectors


Ratio of EPT and Chironomid Abundances



% Contribution of Dominant Taxon


EPT Index

/c\
Community Similarity Index
Ratio of Shredders/Total^ '

Non-
Impaired
rt, (rt rt IB O
O rt *l X O
n n o *o 3

in n 3* n oi
n c n IB 01
n n a. cr
01 IB in t— *
3 ^ rt £1 3-
01 (rt (B 01 IB
3 M. . 3
O> rt O"
t-** IB (B
3" O o n in
01 3 "O- O rt
O" rt l-l
M> 01 M- IB in
rt 3 3 03 t-"
01 O. C M- rt
rt 3 O C
Q. 3 01
A O O • rt
C 3 O >".
01 H. 3 O
M 3 3 CB 3
M. 01 C 01
rt 3 3 I-" rt
>< 0 M. 01 0
• ID rt 3
(B IB
O-
Biological
Slightly
Impaired
0 H- rt o O
rt, 3 3" 0 0
rt 01 3 3
rt O 3 TJ 3
O M 0 C
k-» IB IB In 3
IB n X H- M-
n 01 T3 rt rt
B 3 IB M-><
3 rt O O
rt • rt 3 in
rt, IB rt
rt, o Q.-~ • n
on me
n 3 a. xi n
3 la C IB rt
M • IB n c
M» n

3 "0 o in
n IB i-«
n n *-* n IB
IB n o M- in
01 IB in n in
in 3 in 3-
IB rt 3 rt
(A O IB 3"
• o n in 01
o in 3
3 w ^
rt O (B
M. 3 1-1 X
er IB o -a
c c IB
« IB n
H- H rt
O IB
3 a.
Condition
Moderately
Impaired
3 3 IB
O. rt <
n o IB
• (B
n (n
01 -a
3 IB
rt, IB
O in
n
a OL.
in c
• IB

7> 0
(B
a, 1-1
C 0
n in
rt in
M*
O 0
3 r«
M. 3
3 O
in
PI rt
3


Severely
Impaired
"O rt O *13
1 < rt, (B
IB O C
in 0
IB rt n in
3 01 OP *O
rt X 01 IB
• oi 3 n

in IB
3 in
0 I/l
3— -0
t-1 n
•< rt IB
3- in
rt (B IB
O 3 3

IB 0. •
n o
01 3
3 M. 1-1
rt 3 rt,
01
O rt 3"
« IB M.
09 Q.OQ
W 3*
3 or
H-V; a.
In IB
303
w 3 in
IB M-
O M*
n IB
(A
     (a)  Scoring criteria are generally based on percent comparability to the reference station.
     (b)  Determination of Functional Feeding Group is independent of taxonomic grouping.
     (c)  Community Similarity Indices are used in comparison to a reference station.

-------
   Data Summary Sheet (Figure 6.3-3). Each metric
   result is given a score based on percent comparability
   to a reference station.  Evaluation of biological condi-
   tion is based on comparison to the  reference condition
   (site-specific or reference database)  that is representa-
   tive of the "best attainable" condition. Using this
   approach, metrics can be eliminated if found inap-
   plicable,  without altering the biological classification.
   However,  this integrated assessment approach is
   intended  to remain intact to avoid jeopardizing the
   integrity  of the bioassessment concept.
      Scores are totaled and a Biological Condition Cate-
   gory is assigned based on percent comparability with
   the reference station score. Values obtained may some-
   times be  intermediate  to established ranges and
   require some subjective judgment as to assessment of
   biological condition. In these instances, habitat assess-
   ment, physical  characterization, and water quality data
   may aid in the  evaluation process. An explanation of
   the importance of interpreting  biological data in the
   context of habitat quality is presented in Chapter 8.
      The metrics used to evaluate the benthic data and
   their significance are described below.

   Riffle/Run Sample

   Metric 1.  Species Richness

             Reflects health of the community
          through a measurement of the variety of
          taxa (total  number of genera and/or species)
          present.  Generally increases with increasing
          water quality, habitat diversity, and/or habi-
          tat suitability. Sampling of highly similar
          habitats will reduce the variability  in this
          metric attibutable to factors  such as current
          speed and  substrate type. Some pristine
          headwater  streams may be naturally
          unproductive, supporting only a very
          limited  number of taxa. In these situations,
          organic enrichment may result in an
          increase in  number of taxa (including EFT
          taxa).

Metric 2. Modified Hilsenhoff Biotic Index

             Tolerance values range from 0 to 10,
          increasing  as water quality decreases.  The
          index was developed by Hilsenhoff (1987b)
          to summarize overall pollution tolerance of
          the benthic arthropod community with a
          single value. This index was developed as a
          means of detecting organic pollution in
          communities inhabiting rock or gravel rif-
          fles, and has been modified  for this docu-
          ment to include non-arthropod species as
          well, on the basis of the biotic index used
          by the  State of New York (Bode  1988).
             Although Hilsenhoffs biotic index was
          originally developed for  use in Wisconsin,
          it is  successfully used  by several States and
          should prove reliable for extensive use,
          requiring regional modification in some
          instances. Alternative tolerance classifica-
          tions and biotic indices have also been
          developed by some State agencies (Appen-
          dix C). The formula for calculating  the
          Biotic  Index is:
                         HBI=I
          where
          Xj = number of individuals within a species
          tj = tolerance value of a species
          n = total number of organisms  in the sample

             Although it may be applicable for other
          types of pollutants, use of the HBI in
          detecting non-organic pollution effects has
          not been thoroughly evaluated. The State of
          Wisconsin is conducting a study to evaluate
          the ability of Hilsenhoffs index to detect
          non-organic effects. Winget and Mangum
          (1979) have developed a tolerance classifica-
          tion  system applicable to the assessment of
          nonpoint source impact. Additional biotic
          indices are also  listed in U.S. EPA (1983).

Metric 3.  Ratio of Scraper and Filtering Collector
          Functional Feeding Groups

             The  Scraper and Filtering Collector
          Functional Group ratio reflects the riffle/run
          community foodbase and provides insight
          into the nature of potential disturbance fac-
          tors. The proportion of the two feeding
          groups is important because  predominance
          of a  particular feeding type may indicate an
          unbalanced community responding to an
          overabundance of a particular food source.
          The  predominant feeding strategy  reflects
          the type of impact detected.
             A description of the Functional Feeding
          Group concept can be found in Cummins
          (1973). Genus-level  Functional Feeding
          Group designations for most aquatic insects
          can be found in  Merritt and  Cummins
          (1984).
                                                        6-22

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                                                                       DATA SUMMARY SHEET
o\
Station No.
Station Location
Taxa Richness
FBI (modified)
Functional Feeding Groups
Riffle Community
Scrapers/Filt. Collect.
CPOM Community
Shredders/Total
EPT/Chironomidae
% Contribution (dom. family)
EPT Index
Community Similarity Index
Comments:

















































































                             Figure 6.3-3.  Data Summary Sheet suggested for use in recording benthic data utilized in Rapid Bioassessment Protocol III.

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             The relative abundance of Scrapers and
         Filtering Collectors in the riffle/run habitat
         provides an indication of the periphyton
         community composition and availability of
         suspended Fine Paniculate Organic Material
         (FPOM) associated with organic enrich-
         ment.  Scrapers  increase with  increased
         abundance of diatoms and decrease as
         filamentous algae and aquatic mosses
         (which cannot be efficiently harvested by
         Scrapers) increase. However, filamentous
         algae and aquatic mosses provide good
         attachment sites for Filtering Collectors,
         and the organic enrichment often responsi-
         ble for overabundance of filamentous algae
         provides FPOM utilized by the  Filterers.
             Filtering Collectors are also  sensitive to
         toxicants bound to fine particles and may
         decrease in abundance when exposed to
         sources of such bound toxicants (Cummins
         1987). The Scraper to Filtering  Collector
         ratio may not be a good indication of
         organic enrichment if adsorbing toxicants
         are present. This  situation is often
         associated with  point source discharges
         where certain toxicants adsorb readily to
         dissolved organic matter (DOM) forming
         FPOM during flocculation. Toxicants thus
         become available  to Filterers via FPOM.  In
         these instances the HBI and EPT Index may
         provide additional insight. Qualitative field
         observations on periphyton abundance may
         also be helpful  in interpreting results.

Metric 4.  Ratio  of EPT and Chironomidae
          Abundances

             The EPT and  Chironomidae abundance
          ratio uses relative abundance of these indi-
          cator groups as a measure of community
          balance. Good biotic condition  is reflected
          in  communities having a fairly  even distri-
          bution among all four major groups and
          with substantial representation in the sensi-
         tive-groups Ephemeroptera, Plecoptera, and
         Trichoptera. Skewed populations having a
         disproportionate number of the  generally
         tolerant Chironomidae relative to the more
         sensitive  insect groups may indicate
         environmental stress  (Ferrington 1987).
         Certain species of some genera such as
          Cricotopus are  highly tolerant (Lenat 1983,
          Mount et al.  1984), opportunistic, and  may
          become numerically  dominant in habitats
         exposed to metal  discharges where EPT
          taxa are not abundant, thereby providing  a
         good indicator of toxicant stress (Winner
         et al. 1980). Clements et al. (1988) found
         that mayflies were more sensitive than
         chironomids when exposed  to 15 to 32
         of copper.
             Chironomids tend to become increas-
         ingly dominant in terms of  percent taxo-
         nomic composition and relative abundance
         along a gradient of increasing enrichment
         or heavy  metals concentration (Ferrington
         1987).
             An alternative to the ratio of EPT and
         Chironomidae abundance metric is the Indi-
         cator Assemblage Index (IAI) developed by
         Shackleford (1988). The IAI integrates the
         relative abundances of the EPT taxonomic
         groups and the relative abundances of
         chironomids and annelids upstream and
         downstream of a pollutant source to  evalu-
         ate impairment. The IAI may be a valuable
         metric in areas where the annelid  commu-
         nity may  fluctuate substantially in  repsonse
         to pollutant stress.

Metric 5. Percent Contribution  of Dominant Taxon

             The percent contribution of the numeri-
          cally dominant taxon to the total number  of
          organisms is an indication of community
          balance at the lowest positive taxonomic
          level. (The lowest positive taxonomic level
          is assumed to be genus or species in most
          instances.) A community dominated by rela-
          tively few species would indicate environ-
          mental stress. (If the Pinkham and Pearson
          Similarity Index  is used as  a community
          similarity index for metric number 7, this
         metric may be redundant.)  Shackleford
         (1988) has modified this metric to reflect
          "dominants in common" (DIG) utilizing the
         dominant five taxa at the stations of
         comparison.
             This  DIG approach is based  on the
         original metric used in earlier drafts of this
         RBP document. The  DIG will provide a
         measure of replacement or  substitution
         between  the reference community  and the
         downstream station.  The purpose of the
         modification to "percent contribution of
         dominant taxon"  used in RBP III (and RBP
         II)  is to focus on evenness/redundancy of
         the benthic community regardless of taxa
         composition. Compositional shifts  are mea-
         sured by other metrics  such as the commu-
         nity similarity indices.
                                                    6-24

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Metric 6.  EPT Index

             The EPT Index generally increases with
          increasing water quality. The EPT Index is
          the total number of distinct taxa within the
          orders Ephemeroptera,  Plecoptera,  and
          Trichoptera. This  value summarizes taxa
          richness within the insect orders that  are
          generally considered to be pollution
          sensitive.
             Headwater  streams which are naturally
          unproductive may experience an increase  in
          taxa (including EPT taxa) in response to
          organic enrichment. In  this situation,  a
          "missing genera"  approach may be more
          valuable. Shackleford (1988) uses a "miss-
          ing genera"  metric to evaluate the loss of
          EPT taxa from upstream to downstream to
          avoid the complication  in data interpretation
          resulting from  the addition  or replacement
          of genera.

Metric 7.  Community  Similarity Indices

             Community Similarity Indices are used
          in situations where reference communities
          exist. The reference community can be
          derived through sampling or prediction for
          a region using a reference database. Data
          sources  or ecological data files may be
          available to establish a  reference community
          for comparison. The combined information
          provided through a regional analysis and
          EPA's ERAPT  ecological database (Dawson
          and Hellenthal 1986) may be useful for this
          analysis.  Three of the many similarity indi-
          ces available are discussed below:

          • Community  Loss Index—Measures the
            loss of benthic  species between a refer-
            ence station and the  station of compari-
            son.  The Community Loss Index was
            developed by Courtemanch and Davies
            (1987) and is an index of dissimilarity
            with values  increasing as the degree of
            dissimilarity from the reference station
            increases. Values range from 0 to
            "infinity." Based on  preliminary data
            analysis, this index provides  greater dis-
            crimination than the  following two  com-
            munity similarity indices.
          • Jaccard Coefficient of Community—
            Measures the degree of similarity in taxo-
            nomic composition between two stations
            in terms of taxon presence or absence.
    The Jaccard Coefficient discriminates
    between highly similar collections.
    Coefficient values, ranging from 0 to 1.0,
    increase as the degree of similarity with
    the reference station increases. See
    Jaccard (1912), Boesch  (1977), and U.S.
    EPA (1983) for more detail. The formulae
    for the Community Loss Index and the
    Jaccard Coefficient are
            Community Loss =
                               d-a
         Jaccard Coefficient =
                              a + b+c
    where
 a = number of species common to both
      samples
 b = number of species present in  Sample B
      but not A
 c = number of species present in  Sample A
      but not B
 d = total number of species present in
      Sample A
 e = total number of species present in
      Sample B

    Sample A  = reference station
    Sample B   = station of comparison

 •  Pinkham and Pearson Community
    Similarity  Index—Measures the  degree of
    similarity in taxonomic composition in
    terms of taxon abundances and can be
    calculated  with either percentages or
    numbers. A weighting factor can be
    added that assigns more significance  to
    dominant species. See Pinkham and Pear-
    son  (1976)  and  U.S.  EPA (1983)  for more
    detail. The formula  is
           min  (x.  , x.. )  fx.     x..
Si    _ r 	*a    I"    ia  .   ib   / ,
   •ab     max  (xia, xib)  [xg      xb  /
   where
                          weighting factor
    Xja, X|b = number of individuals in the  ilh
            species in Sample A or B

    Other community similarity indices sug-
 gested by reviewers of this document
 include Spearman's Rank Correlation
                                                   6-25

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          (Snedecor and Cochran 1967), Morisita's
          Index (Morisita  1959), Biotic Condition
          Index (Winget and Mangum 1979), and
          Bray-Curtis Index (Bray and Curtis  1959,
          Whittaker 1952). Calculation of a chi-
          square "goodness of fit" (Cochran 1952)
          may also be appropriate.

CPOM Sample

Metric 8.  Ratio of Shredder Functional Feeding
          Group and Total  Number of Individuals
          Collected

             Also based on the Functional Feeding
          Group concept,  the abundance of the Shred-
          der Functional Group relative to the abun-
          dance of all other Functional Groups allows
          evaluation of potential impairment as indi-
          cated by the CPOM-based Shredder com-
          munity. Shredders are sensitive to riparian
          zone impacts  and are particularly good indi-
          cators of toxic effects when the toxicants
          involved are readily adsorbed to the CPOM
          and either affect the microbial communities
          colonizing the CPOM or the Shredders
          directly  (Cummins 1987).
             The degree of toxicant effects on Shred-
          ders versus Filterers depends on the nature
          of the toxicants  and the organic particle
          adsorption efficiency. Generally,  as the size
          of the particle decreases, the adsorption
          efficiency increases as a function of the
          increased surface to  volume ratio (Hargrove
          1972). As stated in metric 3, water-borne
          toxicants are readily adsorbed to FPOM.
          Toxicants of a terrestrial source (e.g., pesti-
          cides, herbicides) accumulate on CPOM
          prior to leaf fall thus having a substantial
          effect on Shredders (Swift et al. 1988a and
          1988b).  The focus of this approach  is on a
          comparison to the reference community,
          which should  have an abundance and diver-
          sity of Shredders representative of the par-
          ticular area under study. This allows for an
          examination of Shredder or Collector "rela-
          tive" abundance as indicators of toxicity.

   The data collected in the 100-organism  riffle/run
subsample and the CPOM  sample are summarized
according to the information required for each
metric and entered on the Data Summary Sheet
(Figure 6.3-3).
   Each metric result is given a score based on per-
cent comparability to  a  reference station. Scores are
totaled and a Biological Condition Category is
assigned based on percent comparability with the
reference station score (Figure 6.3-3). Values obtained
may sometimes be intermediate to established ranges
and require some subjective judgment as to assess-
ment of biological condition. In these instances, habi-
tat assessment, physical characterization, and water
quality data may aid in the evaluation process.
   For RBP III, four categories of scores are estab-
lished for  the assessment of biological condition. The
power to differentiate four categories for RBP III over
three for RBP  II is derived from additional effort
necessary  for the lowest possible taxonomic identifica-
tions. However, the rationale for metric percentage
ranges is essentially the same as for RBP II. Fig-
ure 6.3-4 outlines the steps that would be taken in a
biological  assessment patterned after Protocol III.
 6.4  RESULTS OF A PILOT STUDY
  CONDUCTED ON THE ARARAT
      AND MITCHELL RIVERS,
           NORTH CAROLINA
             6.4.1 Introduction

   A joint survey was conducted by EA Engineering,
Science, and Technology and North Carolina Division
of Environmental Management  (DEM) on 23-24 Sep-
tember 1986. The objective of this study was to inves-
tigate several methodological questions raised at the
Benthic Rapid Bioassessment Workshop held in July
1986.
   The principal questions were

• Is it necessary to integrate sampling across all
  appropriate habitats at a given site or will sampling
  a single productive habitat (such as a riffle) suffice
  for a general characterization of biological
  integrity?
• Should abundances be characterized as categorical
  estimates for a total sample or as relative abun-
  dances based on a given size subsample? If counts
  on subsamples are preferred, what is the minimum
  count needed to  detect basic  differences among
  stations?
• Can family-level identifications be useful for a site
  prioritization or  is it necessary to identify all organ-
  isms to the lowest taxonomic level?

   The purpose of this research project was to assess
the use of protocols II and III relative to the above
                                                   6-26

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                                               Site-Specific Study
                                              Sampling & Analysis
                    Metric
1.  Taxa Richness'"'
2.  Hilsenhoff Biotic Index (modified)0"
3.  Ratio of Scrapers/Filt. Collectors*3-"0
4.  Ratio of EPT and Chironomid Abundances'3'
5,  %  Contribution of Dominant Taxon'd)
6.  EPT Index'a)
7.  Community Loss Index'e'
8.  Ratio of Shredders/Total'"-10
                        >80%
                        >85%
                        >50%
                        >75%
                        <20%
                        >90%
                        <0.5
                        >50%
                                                                  Biological Condition Scoring Criteria
  60-80%
  70-85%
  35-50%
  50-75%
  20-30%
  80-90%
  0.5-1.5
  35-50%
40-60%
50-70%
20-35%
25-50%
30-40%
70-80%
 1.5-4.0
20-35%
                                                                                                            0
<40%
<50%
<20%
<25%
>40%
<70%
 >4.0
<20%
(a)  Score is a ratio of study site to reference site X 100.
(b)  Score is a ratio of reference site to study site x 100.
(c)  Determination of Functional Feeding Group is independent of taxonomic grouping.
(d)  Scoring criteria evaluate actual percent contribution, not percent comparability to the reference station.
(e)  Range of values obtained. A comparison to the reference station is incorporated in these indices.
                                               BIOASSESSMENT
                     % Comp.
                      to Ref.
                      Score(a)
 Biological Condition
	Category	
Attributes
                      >83%     Nonimpaired
                     54-79%     Slightly impaired
                     21-50%     Moderately impaired
                      <17%     Severely impaired.
                        Comparable to the best situation to be
                        expected within an ecoregion. Balanced
                        trophic structure. Optimum community
                        structure (composition and dominance)
                        for stream size and habitat quality.
                        Community structure less than
                        expected. Composition (species  rich-
                        ness) lower than expected due to loss
                        of some intolerant forms. Percent con-
                        tribution of tolerant forms increases.
                        Fewer species due to loss of most
                        intolerant forms. Reduction in EPT
                        index.
                        Few species present. If high densities
                        of organisms, then dominated by one
                        or two taxa.
                    (a) Percentage values obtained that are intermediate to the above ranges
                        will require subjective judgement as to the correct placement.  Use
                        of the habitat assessment and physiochemical data may be necessary to aid
                        in the decision  process.
                                                Recommendations
   Figure 6.3-4.  Flowchart of bioassessment approach advocated for Rapid Bioassessment Protocol III.

                                                      6-27

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 questions. The purpose was not to disprove or dis-
 credit any sampling techniques or assessment methods
 presently in use. As stated earlier, the benthic rapid
 bioassessment protocols are essentially a synthesis of
 existing methods that  have been employed for some
 time by various States, (e.g., North Carolina, New
 York, and Virginia). This guidance, therefore, is
 meant to provide basic, cost-effective data gathering
 methods for States that (1) have no established bioas-
 sessment procedures,  (2) are looking  for alternative
 methodologies, or (3) may need to supplement their
 existing programs, (not supersede other bioassessment
 approaches that have already been successfully
 implemented). Furthermore, the results of the Ararat
 and Mitchell River Pilot Study should not be viewed
 as full validation of Rapid Bioassessment Protocols II
 and III. Subsequent studies and additional refinement
 in the course of implementation are needed to fully
 validate the procedures presented in this document.
    The Pilot Study was performed in conjunction with
 North Carolina DEM  because their methods were well
 developed and supported by a large database. There-
 fore, results from the  North Carolina DEM biosurvey
 provide the basis of evaluation for resolution of the
 issues listed above.

                 6.4.2  Methods

   The study site was  the Ararat River near Mt. Airy,
North Carolina, located in the Central Appalachian
Ridge and Valley ecoregion. Sampling was conducted
at four stations on the Ararat River an'd one station on
the  Mitchell  River.  The Mitchell River served as a
reference  site representative of excellent biological
condition within the region. Two sites above the  town
of Mt. Airy  were selected to be used  as site-speqific
controls. The station selected as a regional  reference
(Station R) was on the Mitchell River, located near
the  Ararat River in the same county.  A description of
the  biological sampling stations as illustrated in Fig-
ure 6.4-1 are as follows:

  Station 1, Ararat River at NC 104. Station 1
  served as  a control  station and was located near the
  North Carolina-Virginia border. Station  1  was
  established by North Carolina DEM to monitor the
  water quality of the Ararat River above the town of
  Mt.  Airy. Land use in this area was a mixture of
  forestry and agriculture.

  Station 2, Ararat River at NC 52 (Bus.).
   Station 2  provided an alternate control station and
  was located approximately 1 mi above the WWTP
  discharge. Station 2 was established by North Caro-
   lina DEM to assess the impacts of urban runoff
   and any unpermitted discharges. Land use was
   predominantly urban.

   Station 3, Ararat River near SR 2116. Station 3
   was located just above the confluence of Lovills
   Creek and the Ararat River and was several hun-
   dred meters below the WWTP discharge. Land use
   was predominantly urban.

   Station 4, Ararat River at SR 2019. Station 4 was
   a site sampled as part of the North  Carolina DEM
   ambient network on 4 August  1986. Station 4,
   located about 11 miles below the Mt. Airy WWTP
   discharge, was intended  as a primary station for the
   evaluation of recovery. Land use was primarily for-
   estry with some agriculture.

   Station R, Mitchell  River at SR 1419. This "refer-
   ence" station  was established in an  area known to
   have good to excellent water quality. Land use was
   predominantly forestry. Regional reference stations
   are  especially valuable in differentiating between
   the  effects of pollution and natural  seasonal and
   temporal  variation, and in estimating the biological
   potential  of waterbodies  within a region.

6.4.2.1 Field Collections

   Samples  were collected  concurrently by personnel
from North  Carolina DEM and EA Engineering,
Science, and Technology. At each  station, one collect-
ing team from both North Carolina DEM and EA col-
lected two kick-net samples from riffle areas (from
both a fast and a slow current velocity area). In addi-
tion, North  Carolina DEM personnel sampled other
habitats according to their standardized collection
technique utilizing a variety of sampling methods to
collect from all microhabitats present at a station
(Lenat 1988). Methods  included kick net and sweep
net sampling of riffle areas, root masses,  "snags,"
bank areas,  and  macrophyte beds; use  of fine  mesh
samplers into which  invertebrates inhabiting rocks and
logs were washed; sieving of leaf pack and sand  sam-
ples; and  visual  inspection of large rocks and  logs to
collect attached organisms (N.C. DNR and Commu-
nity Development 1983).
   EA samples were sorted in the field, with  all
organisms and sample remains being preserved for
additional analysis. North Carolina samples were also
sorted in the field (according to their standard proce-
dure),  but the organisms collected from the riffle
sample were kept  separate  from those  found in all
other habitats. These samples were preserved in the
event that additional  analysis was needed. Additional
information  collected  included a habitat assessment
and  general  physical characterization of the site (e.g.,
                                                    6-28

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              NC 104
ARARAT RIVER
  SR2019
ARARAT RIVER
   Figure 6.4-1.  Pilot study station locations, Ararat River, North Carolina, September 1986.
               (Taken from 1 October 1986 NC DEM memorandum.)
                                       6-29

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stream depth, width). The EA sampling effort did not
include collection of a CPOM sample. This pilot
study pre-dated  inclusion of the Shredder metric in
RBPs H and III.

6A2.2 Laboratory  Processing

   Samples (vials of organisms) collected by North
Carolina DEM were taken to  their laboratory and
processed according to standard agency procedures
(N.C. DNR and Community Development  1983).
Organisms were identified to genus or species and
tabulated as a total multihabitat sample with  the taxa
composition for the riffle sample presented separately.
Abundances were characterized as categorical esti-
mates of total abundance.
   Samples collected  by EA were taken to the EA
laboratory and processed by a variety of methods to
allow comparisons among data sets.  First,  organisms
picked in the field were  identified to the lowest posi-
tive taxon (species in  most cases). Afterwards, these
organisms were  added to the remainder of the sample
brought in  from the field. Subsampling to  obtain 100
organisms was then performed on the sample using
the methods in Appendix B. After picking and
separating 100 organisms, an additional 100 were
picked and kept separate a second and third time.  All
three 100-organism subsamples (a total of approxi-
mately 300 organisms) were enumerated and identified
as separate entities.

6.4.2.3 Quality Assurance
   Quality assurance  measures were adhered to
throughout the pilot study to ensure  the reliability of
results.  Field collection of samples was conducted in
conjunction with North Carolina DEM personnel;  all
samples being collected simultaneously at a given sta-
tion. Habitat was assessed consistently by the same
individual at all stations. All field efforts were
thoroughly documented.
   All sample processing in the lab  was performed by
the same individual to ensure  consistency in  sorting
and identification. Subsampling was  randomized by
using a random  numbers table. Number of organisms
picked from each  block was recorded to verify ran-
dom distribution and to validate the  Subsampling
procedure.  Taxonomic identification  was separately
documented for  each subsample.

         6.4.3  Bioclassification of
           Stations Based on the
     North Carolina DEM Protocol

   Results  of the North Carolina DEM study are
presented here as described in a memorandum from
Dave Lenat to Steve Tedder dated 1 October 1986
(Lenat  1986). These results form the basis for this
evaluation of the rapid bioassessment protocols.  The
biological condition of the Ararat River is discussed
in reference  to the condition of the Mitchell River
(reference station  "R").
     Although the Ararat River more  than doubles in
   size within the study  area, we would expect only
   minor changes in the composition of the benthic
   community. Depth  and substrate characteristics are
   similar at all sites,  although less sand was observed
   at the Mitchell River station. This may reflect
   fewer nonpoint-source problems in the Mitchell
   River drainage area. Some substrate differences
   may also  be due to differences in soil type.
     Taxa richness values (Table 6.4-1)  indicated
   good-fair  water  quality at Stations 1  and 2. Poor
   water quality was indicated at Station 3, and fair
   water quality at Station 4. Only the Mitchell River
   (Station R) was found to have excellent water qual-
   ity. There was  little indication that urban runoff or
   unpermitted discharges had any effect on the  biota
   of the Ararat River upstream of Station 2. The Mt.
   Airy WWTP discharge eliminated all but the most
   tolerant species, and full recovery appears to take
   over 25 river miles under low flow conditions.
     A Biotic IndexW has been computed for all
   sites, using a numeric abundance of 1 for rare spe-
   cies,  3 for common species, and  10  for abundant
   species. This method will probably give a slightly
   different value  than more quantitative methods, but
   this  type of computation still appears to allow valid
   between-station  comparisons.
     Ranking of stations by the Biotic Index values
   gives results very similar  to the taxa richness
   criteria, i.e.:
                        = 2>4>3.
      Plecoptera (stoneflies) were completely elimi-
   nated at Stations 3  and 4 (Table 6.4-2).
   Ephemeroptera (mayflies) were largely absent at
   Station 3 and sharply reduced at Station 4.
   Trichoptera (caddisflies)  were also eliminated at
   Station 3, but recovered  more quickly than the
   mayflies.
      Patterns of numeric abundance for the major
   groups also  have been very roughly indicated
   (Table 6.4-2) from  both  field notes and lab counts.
   These  data again indicate a strong similarity
   between Stations 1 and 2, but note the  increased
   abundance of some Coleoptera (riffle beetles) at
  WTolerance characterization developed by North Carolina
    DEM.
                                                   6-30

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       TABLE  6.4-1
  BIOCLASSIFICATION  RESULTS  FOR NORTH  CAROLINA  DEM
  MULTIHABITAT  BENTHIC  SAMPLES COLLECTED FROM THE
  ARARAT  (STATIONS 1-4)  AND  MITCHELL  (STATION R)
  RIVERS,  23-24 SEPTEMBER 1986	

                                      Stations
                                                             3
Total  Taxa  Richness

EpT(a)  Taxa Richness

EPT Abundance(b)

Biotic Index

   Numeric Value

   Hilsenhoff Rating

tt Intolerant Taxa

   All
(c)
 # Unique Taxa
                 (d)
 Bioclassification
                     (e)
                    64

                    18

                    92
63

20

75
32

 1

 1
50

11

76
 94

 31

133
2.6
Good
11
14
2
Good-
Fair
2.7
Good
7
11
6
Good-
Fair
3.4
Poor
0
1
2
V.Poor
3.2
Fair
1
3
3
Fair
2.3
V.Good
15
24
24
Excel.
(a)   Intolerant  groups—Ephemeroptera,  Plecoptera, and Trichoptera.
(b)   Rare=l,  common=3, abundant=10,  summed  for all EPT groups.
(c)   Only  those  intolerant  taxa which are common  or abundant  are counted.
(d)   Number  of taxa occuring at only one of the  five  stations,  very
      tolerant species  excluded.
(e)   Based on DEM Taxa Richness Criteria for Piedmont Rivers.
 Station 2. This site also had a greater number of
 "unique" species (defined here as occurring at only
 one of the study sites). These  between-station
 differences are probably due to the presence  of
 Podostemum (riverweed) at Station 2. Unexplained
 factors appear to have reduced or eliminated
 Podostemum growths at Station 1.
   Organic indicator species were generally not
 abundant at Ararat River sites. Only Limnodrilus
 hoffmeisteri was found to be abundant, and only at
 Station 3 (immediately below the WWTP dis-
 charge). Note that another oligochaete taxon, Lum-
. briculidae, was abundant at both Stations 3 and 4.
 This group is  more strongly associated with toxics
 than with organic pollution.
   Toxic indicator species were abundant through-
 out the Ararat River. The presence of Cricotopus
 bicinctus and C. infuscatus gr. at both Stations 1
                             and 2 suggested some upstream toxicity problems.
                             Although these species were abundant at the
                             upstream stations, they were not dominant taxa. At
                             Stations 3 and 4, however, toxic indicator species
                             clearly dominated the benthic macroinvertebrate
                             community. These data indicate that toxic problems
                             are of greater importance in the Ararat River than
                             organic  loading.
                                Other species-level data are also helpful in mak-
                             ing between-station comparisons. These data again
                             indicate comparable water quality at Stations  1 and
                             2. The loss of some intolerant species at Station 2
                             (Heptagenia aphrodite, Helichus, Hydropsyche
                             bronta,  Atherix lanthd) seems to be offset by  the
                             appearance of other intolerant species (Baetisca
                             Carolina, Promoresia elegans). It is also evident
                             that Station 3, just below the Mt. Airy discharge,
                             is occasionally influenced by drift from the
                                           6-31

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          TABLE 6.4-2   TAXA RICHNESS,  BY GROUP,  FOR  SAMPLES COLLECTED BY
                           NORTH CAROLINA DEM FROM  THE  ARARAT (STATIONS 1-4)
                           AND  MITCHELL (STATION  R)  RIVERS

Group
Ephemeroptera
Plecoptera
Trichoptera
Coleoptera
Odonata
Megaloptera
Diptera: Misc.
Diptera: Chiron.
Oligochaeta
Crustacea
Mollusca
Other
Subtotal (EPT)
Total

1
12(a)
2
4
6
3
2
4
23
3
2(a)
13(a)
6 ,
6
2
4
32
4
1
5(a)
3
31
94
  (a)   Dominant groups  (from  lab  counts  and field  notes).
   upstream sites. This explains the presence of spe-
   cies such as Baetis ptuto and Promoresia elegans.
   These species did not persist 11 miles downstream.
      Several tolerant species were most abundant at
   Station 4, about 11 miles below the discharge.
   These taxa included Rheotanytarsus, Hydropsyche
   betteni, Argia,  and Stenacron interpunctatum. Note
   that S.~interpunctatum has replaced the more
   intolerant S. pallidum at this station.

        6.4.4  Selection  of Metrics

   The analysis techniques in this document focus on
the use of metrics for assessment  of various compo-
nents  of benthic macroinvertebrate community struc-
ture and function. Various metrics were evaluated for
use in making a biological  assessment of the benthic
community in the Ararat and Mitchell Rivers. The
metrics evaluated were:
Taxa or species richness—The number of taxa at the
lowest identifiable level.
Percent contribution of the dominant taxon—
Intended as a simple measure of evenness.
Modified Hilsenhoff Biotic Index (HBI)—Integrates
tolerance classification with abundance.
EPT Index—The number of distinct taxa within the
insect orders Ephemeroptera (mayflies), Plecoptera
(stoneflies), and Trichoptera (caddisflies).
Ratio of abundance of EFT organisms and
chironomid larvae—Intended to  measure a shift in
organism dominance as a response to toxicants or
other pollutants.
Community Similarity Indices—The Jaccard Coeffi-
cient, Pinkham and Pearson Index,  weighted Pink-
ham and Pearson Index,  and Community Loss
Index.
Functional Group  ratios—Shredders/Collectors,
Scrapers/Filterers, and Filterers/Gatherers.
                                                 6-32

-------
   Cluster analyses were used as a means of data
evaluation, providing a level of differentiation among
varied data sets independent of the rapid bioassess-
ment technique. The few data acquired by this one
pilot study do not constitute a rigorous analysis, nor
are the results obtained by the cluster analysis
intended  to be a definitive validation of the rapid
bioassessment technique. Hopefully, a larger database
will be available in the future to more adequately
refine the rapid bioassessment metrics and associated
criteria.
   Thirteen metrics were calculated using the
100-organism subsample data collected from the riffle
habitat of the Ararat and Mitchell River stations. The
resulting information was compared using a cluster
analysis (Figure 6.4-2). The relative proximity of met-
rics on the dendrogram, based  on distance between
cluster centroids, was used to determine the unique
information contributed by each metric to an
integrated bioassessment.
    Seven metrics were considered to add some level
of information to the biological assessment, as
denoted by the distance between cluster centroids.
These are Taxa Richness,  Percent Contribution of  the
Dominant Taxon, Ratio of Scraper to Filtering Collec-
tor Functional Feeding Groups, Community Loss
Index, Ratio of EFT and Chironomid Abundances,
Modified HBI, and EPT Index. The other six metrics
appeared to be somewhat redundant to one or more of
these seven selected metrics in terms of contributed
information. These seven metrics, along with an
eighth metric added subsequently to the pilot study,
the Ratio of Shredder Functional Group to Total Num-
ber of Organisms (derived from a CPOM sample),
form the basis  of the integrated analysis advocated in
this  rapid bioassessment approach.  These metrics  are
described in Sections 6.2 and 6.3. The computed data
for the original seven metrics are presented in Tables
6.4-3 and 6.4-4.
   In addition  to cluster analysis, the bioassessment
technique described in Sections 6.2 and 6.3 was used
to evaluate relationships of biological condition among
stations. These results are presented in  Section 6.4.8.

6.4.5  Comparison of Multihabitat  vs.
        Single Habitat Collections

   From the analysis conducted by  North Carolina
DEM, the  stations were ranked from excellent to poor
biological condition  as follows:
                       = 2>4»3
   The double brackets, which indicate a greater
   degree of difference between stations than a single
   bracket, were added to the North Carolina DEM
p p
p
o
i
                   fi
                   cr
                           B   M
                                                                        in
                                                                        M
                §
                o
                                                                                     Ul
                                                                                            m
             Figure 6.4-2. Cluster analysis results for benthic community metrics, based on 100 organism
                         subsamples from riffle samples collected on the Ararat and Mitchell Rivers.
                                                    6-33

-------
              T/BLE  6.4-3
                           METRIC VALUES, PERCENT COMPARISON, AND BIOASSESSMENT SCORES FOR BENTHIC PILOT STUDY RESULTS:
                           100-,  200-,  AND 300-ORGANISM SUBSAMPLE DATA
Metric Value
Station
Metrics
1
2
3
4
R
% Comparison
Station
1 2
3
4
R
Bioassessment Score
Station
1
2
3
4 R
100-ORGANISM SUBSAMPLE
Taxa, Richness
HB I
Scrapers/Filt. Collect.
EPT/Chiron. Abundance
% Contrib. Dom. Taxon
EPT Indox ...
Community Loss Index
Total Score
Biological Condition
26
4.46
0.833
2.45
11.2
12
0.64


26
4.63
0.604
1.47
19.8
13
0.64


11
9.34
0.000
0.00
53.5
0
2.31


34
6.24
0.108
0.55
16.5
12
0.62


34
3.93
1.500
9.28
14.2
14
0


76 76
88 85
56 40
26 16
11 20
86 93


32
42
0
0
54
0


100
63
7
6
16
86


100
100
100
100
14
100


4
6
6
2
6
4
4
32
Slightly
4
4
4
0
4
6
4
26
Slightly
0
0
0
0
0
0
2
2
Sev.
6 6
2 6
0 6
0 6
6 6
4 6
4 6
22 42
Mod. Non
200-ORGANISM SUBSAMPLE
Taxa Richness
HB I
Scrapers/Filt . Collect.
EPT/Chiron. Abundance
% Contrib. Dom. Taxon
EPT Index d
Community Loss Index
Total Score
Biological Condition
32
4.55
0.92
3.07
11.1
15
0.68


32
4 .57
0 .54
1 .28
21 .1
15
0.73


15
9 . 33
0.00
0.00
56 .5
0
2.12


38
6 .06
0 .26
0.59
16.2
13
0.68


41
3 .63
3 .02
13.40
20.1
17
0


78 78
80 79
30 18
23 10
11 21
88 88


36
39
0
0
56
0


93
60
8
4
16
76


100
100
100
100
20
100


4
4
2
0
6
4
4
24
Slightly
4
4
0
0
4
4
4
20
Mod.
0
0
0
0
0
0
2
2
Sev.
6 6
2 6
0 6
0 6
6 4
2 6
4 6
20 40
Mod. Non
300-ORGANISM SUBSAMPLE
Taxa. Richness
HBITb'
Scrapers/Filt. Collect.
EPT/Chiron. Abundance
% Contrib. Dom. Taxon'
EPT index
Community Loss Index
Total Score
Biological Condition
36
4.56
0.84
2.91
10.6
16
0.68


42
4 .60
0.58
1 .39
19.5
18
0.75


19
9.31
0.00
0.00
56.3
0
2 .00


43
6.07
0.20
0.49
16.7
13
0.64


51
3.55
2.96
15.14
24.2
21
0


70 82
78 77
28 19
19 9
11 20
76 86


37
38
0
0
56
0


84
58
7
. 3
17
62


100
100
100
100
24
100


4
4
2
0
6
2
4
22
Slightly
6
4
0
0
4
4
4
22
Slightly
0
0
0
0
0
0
2
2
Sev.
6 6
2 6
0 6
0 6
6 4
0 6
4 6
18 40
Mod. Non
(a)   Adjusted for use with seven metrics.
(b)   Ratio of reference to station of comparison.
(c)   Actual percent contribution evaluated, not percent comparability.
(d)   Range of values evaluated,  not percent comparability.

-------
              TABLE 6.4-4
                           METRIC VALUES, PERCENT COMPARISON, AND  BIOASSESSMENT SCORES  FOR  BENTHIC PILOT STUDY  RESULTS:
                           EA FIELD-SORTED AND FAMILY-LEVEL  IDENTIFICATION DATA
Metric Value
Station
Metrics 1 2 3 4 • R
% Comparison
Station
1 2 3 4 R
Bioas sessment Score
Station
1 234

R
                                                            EA FIELD SORTED
Taxa. richness
H BI
Scrapers/Filt. Collect.
EPT/Chiron. Abundance
% Contrib. Dora. Taxon
EFT Index
Community Loss Index

Total Score

Biological Condition
18
4.26
1.47
12.6
20.0
11
1 .00
30
3.98
1.60
s.oa
12 .0
15
0.53
7
8.33
0.00
0 .00
53.8
0
3 .50
17
5 .41
0 .32
1.96
23.2
10
1 .06
29
4.19
2.15
32.00
10.9
17
0
62
98
68
39
20
65
—
103
105
74
16
12
88
—
24
50
0
0
54
0
—
59
77
15
6
23
59
—
100
100
100
100
11
100
—
4
6
6
2
4
0
4
6
6
6
0
6
4
4
0
2
0
0
0
0
2
2
4
0
0
4
0
4
6
6
6
6
6
6
6
                                                                                                26         32        4     14     42

                                                                                             Slightly  Slightly   Sev.  Mod.   Non
                                                     FAMILY-LEVEL IDENTIFICATION
Taxa.richness
FBI
Scrapers/Filt. Collect.
EPT/Chiron. Abundance
% Contrib. dom. family
EFT Index
Community Loss Index

Total Score

Biological Condition


ct .
e , ,
iy(c|

(d)
12
5.15
0 .83
2.45
19.0
5
0.83
13
5.37
0.60
1.474
34.2
6
0.85
5
9.30
0.00
0.000
70.9
0
3.60
12
6 .14
0.11
0.550
54 .5
4
1 .08
21
4.59
1.50
9.286
34.0
8
0
                                                                57
                                                                89
                                                                56
                                                                26
                                                                19
                                                                62
62
85
40
16
34
75
24
49
 0
 0
71
 0
57
75
 7
 6
54
50
100
100
100
100
 34
100
 3
 6
 6
 3
 6
 0
 3

27

Mod.
3
3
3
0
3
3
3
0
0
0
0
0
0
3
3
3
0
0
0
0
3
6
6
6
6
3
6
6
                                                                                                          18

                                                                                                          Mod.
                                                                                                                 Sev.
                                                                                                                        Sev.
                                                                                                                                39
                                                                                                                               Non
(a)  Adjusted Cor use with seven metrics.
(b)  Ratio of reference to station of comparison.
(c)  Actual percent contribution evaluated, not percent comparability.
(d)  Range of values evaluated, not percent comparability.

-------
   results to provide another level of differentiation of
   biological condition.
   The North Carolina DEM analysis was conducted
by sampling several habitats and making an integrated
assessment focusing primarily on taxa richness and
EFT Index.  These results were used as a basis for
comparison  to results obtained only from the riffle
habitat. Comparison of taxa richness for the multi-
habitat and riffle samples that were analyzed  using the
same method indicates that more taxa were collected
using the multihabitat approach (Figure 6.4-3). The
general trend of taxa richness among stations was
                                               similar. Although the multihabitat approach provides a
                                               distinct separation of benthic diversity among stations,
                                               data obtained from the riffle habitat alone is sufficient
                                               for a discrimination among the stations with regard
                                               to taxa richness.  A comparison of the EPT Index
                                               between  the  multihabitat and riffle  samples was highly
                                               similar at all stations except at Station R  (Fig-
                                               ure 6.4-3).
                                                 The use of combined information from the seven
                                               metrics was evaluated by performing independent clus-
                                               ter analyses and comparing station  relationship results
                                               with those obtained from the North Carolina DEM

40

35

30-

25-

20

is-

le-

 s'
                 NC Multihabitat
                           —  -
                 NC Field Sorted Riffle
                 EA Field Sorted Riffle
                   NC Multihabitat
                   NC Field Sorted Riffle
                    EA Field Sorted Riffle
                                                         Stations

      Figure 6.4-3. Comparison of taxa richness for all field sorted samples collected on the Ararat (Stations 1—4)
                  and Mitchell (Station R) Rivers.
                                                    6-36

-------
approach. Results of the cluster analysis performed on
the EA field-sorted .riffle sample  (Figure 6.4-4)  indi-
cate a station relationship based on similarity of
attributes from the seven metrics. However, the  rela-
tionships of the stations in terms  of biological condi-
tion cannot be determined using only the. clusters.
Therefore, a  knowledge of the biological results
obtained using the North Carolina DEM analysis tech-
nique is used to put the station relationships in  .
perspective.
                        = 2>4»3
   A further analysis was conducted on the riffle
sample by subsampling to 100 organisms  in the
laboratory, identifying and enumerating, and perform-
ing a cluster analysis on the computed metrics. The
results of the clusters  indicate that Stations  1 and 2
are most similar, with Station 4 being next  most simi-
lar to the centroid of 1 and 2. The reference station
was  unlike any of the others, as was Station 3. Rear-
ranging these results in the context of the biological
data to provide an interpretation of impairment, these
results (Figure 6.4-5) also indicate a strong  similarity
with the classification presented by the North Carolina
DEM. The ranking  of stations according  to biological
condition from the laboratory-processed samples using
1.4 -
3 1.2-
1 1-
*
5 0.8-
O
§ 0.6 -
** 0 4 -
S?
S 0.2 -
5
0 -







I









1 2




































4 R 3
Stations
           Figure 6.4-4.  Station cluster analysis results for field sorted riffle samples collected on the Ararat
                        (Station 1-4) and Mitchell (Station R) Rivers.
                     1.4
                     1.2-
                     o,-
                      0-
                                                           4
                                                        Stations
               n
             Figure 6.4-5. Station cluster analysis results for 100 organism subsamples from riffle samples
                          collected on the Ararat (Stations 1-4) and Mitchell (Station R) Rivers.
                                                      6-37

-------
the clusters is:
                        = 2>4»3
   This preliminary analysis of multihabitat versus
single habitat conducted at one site suggests that the
single habitat approach can provide a representative
sample for an evaluation of biological condition.

          6.4.6  Evaluation of the
        100-Organism  Subsample

   To determine if a 100-organism subsample pro-
vides an adequate  estimate of community structure,
comparisons were  made between  100-, 200-, and 300-
organism subsamples. Although comparison of the
cumulative taxa richness  and EFT values for the 100-,
200-, and 300-organism subsamples (Table  6.4-3) indi-
cates that  additional information is gained with each
incremental increase in organism  count, results of  a
cluster analysis of the seven metrics performed on the
300-organism count data (Figure  6.4-6) showed the
same station relationship as that obtained with the
100-organism data  (Figure 6.4-5). The greater sensitiv-
ity demonstrated with the 300-organism data was
subtle and may not warrant the additional time expen-
diture required.
   Laboratory sorting of each 100-organism subsam-
ple was estimated  to require between 1 and 1.5 hours.
If a 300-organism  subsample was used, approximately
3 to 4.5 hours would be necessary to pick the organ-
isms from the sample. The time-savings estimated
for the 100-organism subsample, combined with the
minimal additional information provided with addi-
tional subsamples, supports the contention that a
100-organism subsample is adequate for assessment of
the benthic community. Other researchers have also
found that a 100-organism subsample will provide
sufficient data to detect impact (Nuzzo 1986, Bode
1988, Shackleford 1988).

           6.4.7 Family-Level vs.
       Species-Level Identification

   Additional tabulation was done to determine if
family-level identifications resulted in similar site clas-
sifications.  A cluster analysis was performed on all of
the 13 metrics described in Section 6.4.4 for the
family-level data of the 100-organism riffle sample.
Results obtained with the cluster analysis (Figure 6.4-7)
showed the same relationship in terms of contribution
of the metrics to the  assessment as did the species-
level (lowest taxon) analysis. Therefore, the seven
metrics used for the  lowest taxonomic level assess-
ment were used for the family-level assessment.
   Data for the seven computed metrics using family-
level identification are presented in Table 6.4-4.
Results of station  clusters illustrate a  spatial trend
(Figure 6.4-8) that is the same as that obtained using
the RBP III approach on the  100-organism (lowest
taxon) data (Figure 6.4-5). Stations 1  and 2 were most
similar;  Station 4 clustered next, then R, and finally
3.  However, the bioassessment scheme of the family-
level protocol  is more general than that provided by
Distance between Cluster Centroids
0 O 0 0 — -
oK>*d»OB-*r\>*
I 1 I I 1 I 1 I

1
1 1








1 2 4 R 3
Stations
            Figure 6.4-6. Station cluster analysis results for 300 organism subsamples from riffle samples
                        collected on the Ararat (Station 1—4) and Mitchell (Station R) Rivers.
                                                   6-38

-------
          
         p
          2
          o>
         O
         S
          in
         O
         5
         S
          4»3
   The family-level  data differed  slightly in level of
station similarity compared to that for the species-
level (I00-organism  riffle), which is to be expected
with different taxonomic levels of identification. How-
ever, results indicate that a reasonably good evaluation
                                            can  be obtained with  family-level identifications. The
                                            relative sensitivity of  a family-level identification effor.
                                            is sufficient for a prioritization or site ranking pro-
                                            tocol that would differentiate between non-impaired,
                                            moderately impaired,  and severely  impaired
                                            conditions.

                                                 6.4.8  Integrated Bioassessment

                                               A summary of the  bioclassification scheme for the
                                            stations that was derived  from the cluster analysis per-
                                            formed on all of the data sets and the North Carolina
                                            DEM analysis technique  is presented  in Table 6.4-5.
                                                     6-39

-------
           TABLE 6.4-5
 SUMMARY  OF THE  BIOCLASSIFICATION DERIVED FROM
AN  ANALYSIS OF  SAMPLES COLLECTED  FROM  THE
ARARAT  AND  MITCHELL RIVERS
                         Data  Set
                                            Bioclassification
  Cluster Analysis  (Based  on NC DEM Classifications)

    NC multihabitat; NC analysis

    EA field-sorted riffle; RBP III analysis

    100-organism riffle; RBP  III  analysis

    300-organism riffle; RBP  III  analysis

    100-organism riffle; RBP  II (family)  analysis
R
R
R
R
R
»
»
»
»
»
1
1
1
1
1
= 2
= '2
= 2
= 2
= 2
>
>
>
>
>
4
4
4
4
4
»
»
»
»
»
3
3
3
3
3
  Bioassessment  Technique

     NC multihabitat; NC analysis

     EA field sorted riffle; RBP III analysis

     100-organism riffle; RBP  III  analysis

     300-organism riffle; RBP  III  analysis

     100-organism riffle; RBP  II (family)  analysis
                                           R » 1  = 2  > 4 »  3

                                           R>1 = 2>4>3

                                           R>1 = 2>4»3

                                           R»l  = 2>4>3

                                           R>1 = 2>4  = 3
Very little variation existed in the relationship of Sta-
tions R, 1, and 2,  whereby R was always of greater
quality than Station 1, and essentially the same quality
existed between Stations 1 and 2. In addition, the
orientation of the stations in terms of biological condi-
tion was the same  for all data sets. The subjectivity in
these analyses exists in the fact that some judgment
has to be used in interpreting the biological relation-
ships from the station similarity  information illustrated
by the dendrograms of the cluster analysis.  It  is possi-
ble that the close proximity of Station 4 to the cen-
troid of Stations 1  and 2 could indicate equality rather
than a slightly lower quality. This situation  occurred
particularly in the  data sets of the 300-organism riffle
subsample and the multihabitat RBP III analysis.
   Using the scoring criteria described for RBP II
(Section 6.2) and RBP III (Section 6.3), the bioassess-
ment metrics were calculated. The bioclassification
for the 100-organism subsample  species-level identifi-
cation resulted in Station R being classified as non-
impaired, Stations  1 and 2 as slightly impaired, Sta-
                        tion 4 as moderately impaired, and Station 3 as
                        severely impaired (Table 6.4-3). Therefore,
                        R>1 =2>4»3. Bioclassification for the
                        300-organism subsample resulted in a classification
                        similar to that of the 100-organism count samples.
                        The station relationship results based on biological
                        condition  using the bioassessment approach are not
                        unlike those obtained using the cluster analysis
                        (assuming the biological condition as identified by
                        NC DEM) on the same data sets. However, the
                        amount of data was limited for an adequate cluster
                        analysis. Station ranking based on results  of the field-
                        sorted riffle sample is similar to that resulting from
                        the North Carolina DEM multihabitat bioclassifica-
                        tion. Although differences among stations were more
                        conservative using the RBP approach, these are the
                        same station trends observed in the cluster analysis for
                        this data set (Table 6.4-5).
                           The  family-level bioclassification results suggest an
                        orientation slightly different from that obtained with
                        the  100-organism (species-level) sample (Table 6.4-4).
                                               6-40

-------
Both the species-level  bioassessment and the station
cluster for the family-level data indicated that Station
3 .is different from Station 4,  which reflects the lesser
sensitivity associated with the family-level identifica-
tion used in RBP  II (Table 6.4-5). The difference
between  family-level bioclassification (moderate
impairment) and species-level  bioclassification (slight
impairment) at Stations  1 and 2  is attributable to the
fact that  the RBP  II classification scheme is based  on
only three levels of impairment as opposed to the four
levels used in RBP III.
   The bioassessment technique appears to be more
conservative than  the clustering technique,  which may
be beneficial from  a water quality management point
of view.  Subtle differences in structure and function
will  be regarded  as rationale for further confirmative
study,  to ascertain  the significance of complex impair-
ment problems.  If  an evaluation of biological  condi-
tion  was based on  a straight percent-of-refere.nce, a
slightly different scenario might be obtained.  A
greater differentiation  between Stations R and 1  and
between  Stations 3 and 4 would be one outcome.
However, based on this single pilot study, the ranges
of biological condition are necessarily conservative.
With a good reference database, these ranges  can be
modified, making them either more or less protective.
                                                      6-41

-------
                 7.  FISH BIOSURVEY  AND  DATA  ANALYSIS
   Two levels of fish biosurvey analyses are
presented: Rapid Bioassessment Protocol IV consti-
tutes a questionnaire approach where local and State
fisheries experts are canvassed for existing data and
information; Rapid Bioassessment Protocol V consists
of collecting fish at selected sites for biosurvey anal-
yses. The data analysis used in RBP V is based on
the IBI  (Karr et al. 1986) and the IWB (Gammon
1980). This document only provides an overview of
the IBI  and IWB and their conceptual foundations.
Effective use of RBP V requires information presented
in Karr  et al. (1986) and Gammon (1980).  Sample
field and data sheets are presented as guidance.
   Pilot studies based on use of the fish biosurvey
(RBP V) have been published. An overview of two of
these studies is presented in Section 7.3.
     7.1 RAPID BIOASSESSMENT
         PROTOCOL IV-FISH
   The intent of RBP IV is to serve as a screening
tool and to maximize the use of existing knowledge of
fish communities.  The questionnaire polls State fish
biologists and university ichthyologists- believed
knowledgeable about the fish assemblages in stream
reaches of concern. The proposed questionnaire (Fig-
ure 7.1-1)  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 questionnaires include  inaccuracy
caused by hasty responses, a desire to report condi-
tions as better or worse than they are, and insufficient
knowledge. The questionnaire provides a qualitative
assessmenrof"a~ large number of waterbodies quickly
and inexpensively. Its quality depends on the survey
design (the number and location of waterbodies), the
questions presented, and the knowledge  and coopera-
tion of the  respondents.
   This document 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.
   7.1.1  Design of Fish Assemblage
           Questionnaire Survey

   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 waterbody
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 modifica-
tions, and diversions, and have diverse habitats, com-
plex  bottom substrate, considerable  instream cover,
and a wide buffer of natural riparian vegetation.
   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
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 topo-
graphic 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.
   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 fiscal year. The questionnaire should be accom-
panied by a personalized cover letter written on offi-
cial stationary, and closed by an official title below
the signature. A  stamped, self-addressed return enve-
lope increases the response rate.  Materials mailed first
or priority class  are effective; special  delivery and
certified letters are justified in follow-up mailings. Tel-
ephone contact is advisable after three follow-up
notes.

          7.1.2  Response Analysis

   Questionnaire response should provide the follow-
ing information:

1. The  integrity  of the fish community
                                                   7-1

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                    FISH ASSEMBLAGE QUESTIONNAIRE
INTRODUCTION
This questionnaire  is part of an effort  to assess  the biological health
or integrity of  the flowing vaters of  this state.  Our principle focus is
on the biotic health of the designated vaterbody 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  vaterbody 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 waterbody.  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 aware of
someone who is familiar with the waterbody, please give this person's
name, address, and  telephone number in the space provided below.
Vaterbody code

Vaterbody name
Vaterbody location  (also see map)

    State 	  County 	    Long/Lat	

    Ecoregion 	

Vaterbody size


    Stream  (<1  cfs,  1-10 cfs, >10 cfs)
              r

(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 waterbody
   size and ecoregion.

3  Intolerant species absent, considerably  fewer species and individuals
   than expected  for that waterbody size  and ecoregion,  older age  classes
   of top carnivores rare, trophic structure skewed toward omnivory.
 Figure 7.1-1.  Fish assemblage questionaire for use with Rapid Bioassessment Protocol IV.
                                     7-2

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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 fish 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

        5  -4   3   2   1   0                          '	'

2.  Rank the conditions of the reach 10 years ago

        543210

3.  Given present trends, hov will the reach rank 10 years from now?

        543210                               ....

4.  If the major human-caused limiting factors were eliminated, how
    would the reach rank 10 years from now?
(Complete each subsection by circling the single most appropriate
limiting factor and probable cause.)
Subsection 1—Water Quality

Limiting factor

5   Temperature too high
6   Temperature too low
7   Turbidity
8   Salinity
9   Dissolved oxygen
10  Gas supersaturation
11  pH too acidic
12  pH too basic  •
13  Nutrient deficiency
14  Nutrient surplus
15  Toxic substances
16  Other (specify below)
17  Not limiting
Probable cause

18  Primarily upstream
19  Within reach
20  Point source discharge
21    Industrial
22    Municipal
23    Combined sewer
24    Mining       '
25    Dam release
26  Nonpoint source discharge
27    Individual sewage
28    Urban runoff '•
      Landfill leachate
      Construction
      Agriculture
      Feed lot
      Grazing
      Silviculture
      Mining
29
30
31
32
33
34
35
36  Natural"
37  Unknown
38  Other (specify below)
                          Figure 7.1-1. (Cont.).
                                  7-3

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Subsection  2--Vater 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    Mining
53  Natural
54  Unknown
55  Other (specify below)
Subsection 3--Habitat Structure

Limiting factor

56  Excessive siltation
57  Insufficient pools
58  Insufficient riffles
59  Insufficient shallows
60  Insufficient concealment
61  Insufficient reproductive
    habitat
62  Other (specify below)


63  Not limiting




Subsection 4—Fish Community

Limiting factor

76  Overharvest
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  Mining
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—Major Limiting Factor
                                    93  Water quality
                                    94  Water quantity
                                    95  Habitat structure
                                    96  Fish community
                                    97  Other (specify)
Your name  (please print)
                      Figure 7.1-1.  (Cont.).
                              7-4

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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 waterbody 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
   The questionnaire data are most effectively entered
and analyzed by using a microcomputer and interac-
tive database 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.
RBP IV allows considerable flexibility.
    7.2  RAPID BIOASSESSMENT
          PROTOCOL  V—FISH
   Rapid Bioasssessment Protocol V (RBP V) is a
rigorous approach similar to macroinvertebrate
RBP III in accuracy and effort, but focuses on the
fish community.  RBP  V involves careful, standardized
field collection, species identification and enumera-
tion, and community analyses using biological indices
or quantification of the biomass and numbers of key
species. The RBP V survey yields an objective, dis-
crete measure of the health of the fish community that
usually can be completed onsite by qualified fish biol-
ogists (difficult species identifications may require
laboratory confirmation). Data provided by RBP V
can serve  to assess use attainment, develop biological
criteria, prioritize sites for further evaluation, provide
a reproducible impact assessment, and assess fish
community status and trends. RBP V is based primar-
ily on the Index  of Biotic Integrity (IBI), a fish com-
munity assessment approach developed  by Karr (1981).
A more detailed  description  of this approach  is
presented  in Karr et al. (1986)  and Ohio EPA (1987b).
Regional modifications and applications are described
in Hughes and Gammon (1987), Leonard and Orth
(1986), Steedman (1988), Wade and Stalcup (1987),
and Miller et al. (1988a).
       7.2.1 Field Survey  Methods

   RBP V involves field evaluation of the same phys-
icochemical and habitat characteristics as RBPs I, II,
and III (Figures  5.1-1 and 5.2-1), a similar impairment
assessment (Figure 7.2-1), and a fish community
biosurvey. Because it provides critical information for
evaluating the cause and  source of impairment, the
habitat and physical characterization (described in
Chapter 5 of this document) are essential to RBP V.
The approach for conducting a RBP V site-specific
fish community  analysis is based on the use of the IBI
(Figure 7.2-2).

7.2.1.1  Sample Collection
   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
RBP V. However,  pilot studies may indicate the need
for different or multiple gear.
   The fish community biosurvey  data are designed to
be representative of the fish community at  all  station
habitats,  similar to the "representative qualitative sam-
ple" 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 pro-
vides a realistic  sample of fishes likely to be encoun-
tered  in the waterbody. Sampling procedures effective
for  large rivers are described in Gammon (1980),
Hughes and Gammon (1987), and Ohio  EPA (1987b).
   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 effec-
tive if the water is especially clear  and there is little
cover (Reynolds  1983). 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.
   The RBP V fish community assessment 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
                                                    7-5

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Field crew electrofishing with a pram-towed unit.
                       7-6

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                              IMPAIRMENT ASSESSMENT SHEET
       1.   Detection of impairment:   Impairment  detected
                                     (Complete Items 2-6)
             No impairment
                detected
              (Stop here)
       2.   Biological impairment indicator:

            Pish

            	 sensitive species reduced/absent
            	 dominance of tolerant  species
            	 skewed trophic structure
            	 abundance reduced/unusually  high
            	 biomass reduced/unusually.high
            	 hybrid or exotic abundance   '
                  unusually high
            	 poor size class representation
            	 high incidence of anomalies

       3.   Brief description of problem: 	
  Other aquatic communities

  	 Macroinvertebrates
  	 Periphyton
  	 Macrophytes
           Year and date of previous surveys:

           Survey data available in: 	
       i>.   Cause (indicate major cause):   organic enrichment   toxicants   flow

                                        •  sediment   temperature   poor habitat

                                          other
       5.   Estimated areal extent' of problem (m )  and length of stream reach

           affected (m) where applicable:	-

       6.   Suspected source(s) of problem
             point source

             urban runoff

             agricultural runoff
             silvicultural runoff
             livestock
             landfill
mine
dam or diversion

channelization or snagging

natural
other
unknown
       Comments:
Figure 7.2-1.  Impairment Assessment Sheet for use with fish Rapid Bioassessment Protocol V.
                                        7-7

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                                         I    Select a Site

                                                  I
                                     Identify Regional Fish Fauna
                                                 I
                        Assign Species to Trophic, Tolerance, and Origin Guilds
                         Assess Available Data for Metric Suitability and Stream
                                            Size Patterns
                                                 _L
                             Develop Scoring Criteria from Reference Sites
                                      Quantitatively Sample Fish
                                                 I
                          List Abundances of Species, Hybrids, and Anomalies
                                                 _L
                                   Calculate and Score Metric Values
METRIC SCORES (IBI)
Scoring Criteria'"'

1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
Metric
Number of native fish species
Number of darter or benthic species
Number of sunfish or pool species
Number of sucker or long-lived species
Number of intolerant species
Proportion of green sunfish or tolerant
individuals
Proportion omnivorous individuals
Proportion insectivores
Proportion top carnivores
Total number of individuals
Proportion hybrids or exotics
Proportion with disease/anomalies
5
>,67%
>67%
>67%
>67%
>67%
<10%
<20%
>45%
>5%
>67%
0%
3
33-67%
33-67%
33-67%
33-67%
33-67%
10-25%
20-45%
20-45%
1-5%
33-67%
0-1%
<1% 1-5%
'"'Metrics 1-5 are scored relative to the maximum species richness
Metric 10 is drawn from reference site data.
1
<33%
<33%
<33%
<33%
<33%
>25%
>45%
<20%
<1%
<33%
>1%
>5%
line.
                                INDEX SCORE INTERPRETATION'"'
                  IBI
                58-60

                48-52
                28-34
                 12-22
Integrity Class
Excellent

Good


Fair

Poor


Very Poor
                                                            Characteristics
Comparable to pristine conditions,
exceptional assemblage of species
Decreased species richness,
intolerant species in particular;
sensitive species present
Intolerant and sensitive species
absent; skewed trophic structure
Top carnivores and many expected
species absent or rare; omnivores and
tolerant species dominant
Few species and individuals present;
tolerant species dominant; diseased
fish frequent
                  (a'From Karr et al. 1986; Ohio EPA 1987.
                                          Recommendations

Figure 7.2-2.  Flowchart of bioassessment approach advocated for Rapid Bioassessment Protocol V.

                                                 7-8

-------
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) recom-
mended 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 collection (e.g., sturgeon).
The intent of the sample (as with the entire protocol)
is  to obtain a representative estimate of the species
present, and their abundances, for a reasonable
amount of effort.
   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 compari-
son of fish community status among sites.  Major habi-
tat 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 con-
siderably with the gear  used and the size and com-
plexity of the site.
   Atypical conditions,  such as high flow,  excessive
turbidity  or turbulence,  heavy rain, drifting leaves,  or
other unusual conditions, that affect sampling effi-
ciency, are best avoided. Glare, a frequent problem, is
reduced by wearing polarized glasses during sample
collection.

7.2.1.2  Sample Processing

   A field collection data sheet (Figure 7.2-3) is com-
pleted 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  num-
bers and weights and the incidence of  external anoma-
lies are recorded for each group.  Reference specimens
of each species from each site are preserved in
10 percent formaldehyde, the jar labeled, and the col-
lection 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 weighed 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  mor-
tality  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 (1987b).

    7.2.2  Data Analysis Techniques

   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 7.2-1). A
preliminary  judgment on the  presence of biological
impairment  is particularly important if RBP IV is not
used prior to RBP V.
   Data can be analyzed using the IBI (or individual
IBI metrics), the IWB (Gammon  1980), and multivari-
ate  statistical techniques  to determine community
similarities.  Detrended correspondence analysis (DCA)
is a useful multivariate analysis technique for revealing
regional community patterns  and  patterns among mul-
tiple sites. It also demonstrates assemblages with com-
positions differing from others  in the region or reach.
See Gauch (1982) and Hill (1979) for descriptions of,
and software for, DCA. 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. Illinois EPA has developed
software to assist professional aquatic  biologists in cal-
culating IBI values  in Illinois streams  (Bickers et al.
1988). (Use of this software outside Illinois 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.
   The IBI  is a broadly-based index firmly grounded
in fisheries  community ecology (Karr 1981; Karr et al.
1986). The IBI incorporates zoogeographic, ecosys-
tem, community, population, and individual perspec-
tives.  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
                                                     7-9

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                           FISH FIELD COLLECTION DATA SHEET
                                                                   page
                                                                           of
                                           Date .
Drainage	
Sampling Duration  (rain)
Sampling Distance  (m)
Habitat Complexity/Quail
Weather
Gear Used
Comments 	
Fish (preserved)  Number  of  Individuals
      Sampling Area
ty (excellent   good
  Crew 	
poor   very
                                                       _
                                                      fair
                                   Flow  (flood  bankfull  moderate  lov)
                                   Gear/Crev  Performance
poor)
                                                  Number  of  Anomalies
      Genus/Species
                                 Adults
                   Juveniles
                               No.
                                      Ut.
                                                  No.
                                                          tft.
      Anomalies
           No.
                                                                            (*)
      (*) Discoloration,  deformities,  eroded  fins,  excessive  mucus,  excessive

          external  parasites,  fungus,  poor condition,  reddening,  tumors,

          and  ulcers
Figure 7.2-3.  Fish Field Collection Data Sheet for use with Rapid Bioassessment Protocol V.
                                         7-10

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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).
   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 com-
position 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 (Sec-
tion 7.2.2.1).
    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 quantita-
tive standards for discriminating fish  community con-
dition. Judgment is involved in choosing the most
appropriate population or community element that is
representative of each metric and .in setting the scor-
ing 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 expectations 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 Appendix D. Additional classifi-
cations 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  southeastern
and southwestern United States and its widespread use
by  water resource agencies may result in  further
modifications.  Past modifications have occurred (Sec-
tion 7.2.2.1;  Miller et al. 1988a) without  changing the
IBI's basic theoretical foundations.  Sample calcula-
tions of the IBI are given in Section 7.3.
    The steps in calculating the IBI (Figure 7.2-2) are
explained below:

 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.
2.  Develop scoring criteria for each IBI metric. Maxi-
    mum species  richness (or density)  lines are devel-
   oped from a reference database.
3.  Conduct field study and  identify fish; note anoma-
   lies,  eroded fins, poor condition, excessive mucous,
   fungus, external parasites, reddening, lesions, and
   tumors. Complete field data sheets.
4.  Enumerate and tabulate number of fish  species and
   relative abundances.
5.  Summarize site information for each IBI metric.
6.  Rate each IBI metric  and calculate total IBI score.
7.  Translate total IBI score  to one of the five integrity
   classes.
8.  Interpret data in the context of the habitat assess-
   ment (see Chapter 8). Individual metric analysis
   may  be necessary to ascertain specific trends.

   The IBI is based on an integrated analysis of the
metrics. However, individual IBI metrics may serve as
separate variables to aid in  data interpretation. Com-
parison of commonly-occurring and dominant species
are revealing, especially  when  related to their ecologi-
cal requirements and tolerances. Larsen et al. (1986)
and Rohm et al. (1987) provide examples of such
regional characterizations of common and  abundant
species. The IWB (Gammon 1980; Hughes and Gam-
mon 1987) incorporates two abundance and two diver-
sity 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=0.5.1n N + 0.5 In B+H'N+H'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 and J.R. Gammon (Gammon  1989) found
that subtracting highly tolerant species from the num-
ber and biomass variables increases sensitivity of the
index in degraded environments (Ohio EPA 1987b).
   If the  size of a particular fish population (e.g.,
trout or bass species) is  of  concern,  it can be esti-
mated with known confidence limits by several
methods.  One of the most popular approaches is the
removal method (Seber and LeCren 1967;  Seber and
Whale  1970,  Seber 1982). This approach assumes a
closed population,  equal probability of capture for all
fish, and  a constant probability of capture from sam-
ple 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 = C,2/(C,-C2)
where  C,  and C2 are the number of fish captured in
                                                     7-11

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the first and second samples, respectively.
In the three sample cases, population size is estimated
by
                                            J/2
      N =
6X2 - 3XY - Y? 4- Y(Y2 + 6XY - 3X?)
              18(X-Y)
where X=2C,+C2, and Y = C,+C2 + C3.

   Many methods are available to calculate population
statistics from removal data including regression, max-
imum likelihood, and  maximum weighted-likelihood.
Public-domain software is available to assist in cal-
culating these and other fisheries population statistics
(Van  Deventer and Platts 1989).

7.2.2.1 Description of BBI Metrics

   The IBI serves as an integrated analysis because
individual metrics may differ in their relative sensitiv-
ity to various levels of biological  condition. A
description and  brief rationale for each of the  12 IBI
metrics is outlined below. The original metrics
described by Karr (1981) for Illinois streams (under-
lined) are  followed by substitutes  used in or proposed
for different geographic regions and stream sizes.
Because of zoogeographic differences, different fami-
lies or species are evaluated in different regions,  with
regional substitutes occupying the same general habitat
or niche. The source for each  substitute is footnoted
below. Table 7.2-1' 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 7.2-1 as an example of the assessment approach
for evaluating fish community  condition.

Species Richness and Composition Metrics
   These  metrics assess the species richness compo-
nent  of diversity and the health of the major taxo-
nomic 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 and their substitutes, 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 7.2-4). A detailed description of
these methods can be found in Fausch et al.  (1984),
Ohio EPA (1987b), and Karr et al (1986).
Metric 1.   Total number of fish species(l,4,5). Sub-
           stitutes: Total number of native fish spe-
           cies(2,8), and salmonid  age classes(6)
              This number decreases with increased
           degradation; hybrids and introduced spe-
           cies are not included. In coldwater streams
           supporting  few fish species, the age classes
           of the species found represent the suitabil-
           ity 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.

Metric 2.   Number and identity of darter spe-
           cies(l). 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(S),  sculpin and darter species(8)
              These species are sensitive to degrada-
           tion 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 m^
           (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.

Metric 3.   Number and identity of sunfish spe-
           cies(l). Substitutes: Number and identity
           of cyprinid  species(2,4),  water column  spe-
           cies(3,4), salmonid species(4), headwater
           species(S),  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 salmor.ids  are  important
           sport species. The sunfish metric works
                                                     7-12

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                                TABLE 7.2-1  REGIONAL VARIATIONS OF IBI METRICS(a)
                                                     New               Central    Colorado    Western  Sacramento-
	Variations in IBI Metrics	    Midwest  England  Ontario  Appalachia  Front Range  Oregon   San Joaquin

1.  Total Number of Species                  XX                 XX                     X

    # native fish species ,, ,.                                  X                                 X
    # salmonid age classes                                                                       XX

2.  Number of Darter Species                 X                           XX

    # sculpin species                                                                            X
    # benthic insectivore species                      X
    # darter and sculpin species      (.^                      X
    # salmonid yearlings (individuals)                                                           XX
    % round-bodied suckers                   X
    # sculpins (individuals)                                                                                X

3.  Number of Sunfish Species                X                                        X

    # cyprinid species                                                                           X
    # water column species                             X
    ft sunfish and trout species                                X
    # salmonid species                                                                                      X
    # headwater species                      X

4.  Number of Sucker Species                 XX                                         X

    # adult trout species^ '                                                                     X          X
    # minnow species                         X                                        X
    ft sucker and catfish species                               X
(a)  Taken from Karr et al. (1986), Hughes and Gammon (1987), Miller et al. (1988a), Miller et al. (1988b In
     Review), Ohio EPA (1987b), Steedman (1988).
(b)  Metric suggested by Moyle or Hughes as a provisional replacement metric in small western salmonid streams.
(c)  An optional metric found to be valuable by Hughes and Gammon (1987).

Note:   X = metric used in region.  Many of these variations are applicable elsewhere.

-------
                                                TABLE 7.2-1  (Cont.)
      Variations in IBI Metrics
                                                     New               Central    Colorado    Western   Sacramento-
                                          Midvest  England  Ontario  Appalachia  Front Range  Oregon    San  Joaquin
5.  Number of Intolerant Species

    tt sensitive species
    tt amphibian species
    presence of J)rook trout

6.  % Green Sunfish

    % common carp
    % white sucker
    % tolerant species
    % creek chub
    % dace species
7.  % Omnivores

    % yearling salmonids1

8.  % Insectivorous Cyprinids
,(b)
    % insectivores
    % specialized insectivores
    # juvenile trout
    % insectivorous species

9.  % Top Carnivores

    % catchable salmonids
    % catchable wild trout
    % pioneering species
    Density catchable wild trout
                                             X

                                             X
                                                                                      X          X

                                                                                      X          X
                                                                                                            X

                                                                                                            X

-------
                                                TABLE 7.2-1  (Cont.)
	Variations in IBI Metrics

10. Number of Individuals

    Density of individuals

11. % Hybrids

    % introduced species
    % simple lithophils
    # simple lithophilic species
    % native species
    % native wild individuals

12. % Diseased Individuals

13. Total Fish Biomass
           New               Central    Colorado    Western  Sacramento-
Midwest  England  Ontario  Appalachia  Front Range  Oregon   San Joaquin
             X

             X
   X
   X
                                                       X

                                                       X
                                                                  X
                                                                  X

-------
     40
%

-------
          for most Mississippi Basin streams, but
          where sunfish are absent or rare, other
          groups are used. Cyprinid species are used
          in cool water western streams; water
          column species  occupy the same niche in
          northeastern streams; salmonids are suit-
          able in coldwater streams; headwater spe-
          cies 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 depen-
          dent 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.

Metric 4.  Number and identity of sucker spe-
          cies(l). Substitutes: Number of adult trout
          species(6),  number of minnow species(5);
          and number of  suckers and catfish(S)
              These species are sensitive to physical
          and chemical habitat degradation and  com-
          monly comprise most  of the fish biomass
          in streams.  All  but the minnows are long-
          lived species and provide a multiyear
          integration  of physicochemical 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 spe-
          cies/waterbody  size relationships.

Metric 5.  Number and identity of intolerant spe-
          cies(l). Substitutes: Number and identity
          of sensitive species(S), amphibian spe-
          cies(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 a disturbance. Species classified
          as intolerant or sensitive should only  rep-
          resent the 5-10  percent most susceptible
          species, otherwise this becomes a less dis-
          criminating metric. Candidate species are
           determined by examining regional ichthyo-
           logical 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  spe-
           cies 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.  Steed-
           man (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.

Metric 6.   Proportion of individuals as green  sun-
           fish (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 distin-
guishes low from moderate quality waters. These spe-
cies  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 suckers 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 weighting the met-
ric 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.

Trophic Composition Metrics
   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 differ-
ent situations can yield similar results. The trophic
                                                     7-17

-------
composition metrics offer a means to evaluate the shift
toward more generalized foraging that typically occurs
with increased degradation of the physicochemical
habitat.

Metric 7.  Proportion of individuals as omni-
           vores(l,2,3,4,5,8).  Substitutes: Proportion
           of individuals as yearlings(4)
              The percent of omnivores in the com-
           munity increases as  the physical and chem-
           ical habitat deteriorates.  Omnivores are
           defined as species that consistently feed  on
           substantial proportions of plant and animal
           material. Ohio EPA  (1987b) excludes sensi-
           tive filter feeding species such as paddle-
           fish 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.

Metric 8.  Proportion of individuals as insec-
           tivorous cyprinids(l). Substitutes:  Propor-
           tion of individuals as insectivores  (2,3,5),
           specialized insectivores(4), and insec-
           tivorous species(S);  and  number of juvenile
           trout(4)
              Insectivores or invertivores are  the
           dominant trophic guild of most North
           American surface waters. As the inver-
           tebrate  food source decreases in abundance
           and diversity due to physicochemical habi-
           tat 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.

Metric 9.  Proportion of individuals as top carni-
           vores(l,3,8). Substitutes: Proportion of
           individuals as catchable salmonids(2),
           catchable wild trout(4), and pioneering
           species(5)
              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 pisci-
           vores,  such  as creek chub and channel cat-
           fish, are not included. In trout streams,
           where  true pisctvores 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 spe-
           cies are  used by  Ohio EPA (1987b) in
           headwater streams typically lacking
           piscivores.

 Fish Abundance and Condition Metrics

    The last three metrics indirectly evaluate popula-
 tion 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.

 Metric 10.  Number of individuals in sam-
           ple(l,2,4,5,8).  Substitutes: Density of
           individuals(3,4)
              This metric evaluates population abun-
           dance and  varies  with region and stream
           size for small  streams. It is expressed as
           catch per unit effort,  either by area, dis-
           tance, 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 as a three, and
           less than 4 as  a one.  Unusually  low num-
           bers 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 addi-
           tional metric.

Metric 11.  Proportion of individuals  as hybrids(l).
           Substitutes: Proportion of individuals as
           introduced  species(2,4), simple  litho-
           phils(5); and number of simple lithophilic
           species(5)
              This metric is an estimate of reproduc-
           tive isolation or the suitability of the habi-
           tat 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
                                                     7-18

-------
          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 sub-
          strates without parental care. Hughes and
          Gammon (1987) and Miller et al. (1988a)
          propose  using percent introduced indi-
          viduals.  This metric is a direct measure of
          the loss  of species segregation between
          midwestern and western fishes that existed
          before the introduction of  midwestern spe-
          cies to western rivers.

Metric 12. Proportion of individuals with disease,
          tumors, fin damage, and skeletal
          anomalies(l).
              This  metric depicts the health and con-
          dition 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 concen-
          trated. They are  excellent  measures of the
          subacute effects of chemical  pollution and
          the aesthetic value  of game and nongame
          fish.
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.

   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 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. This
has been done 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 7.2-3), 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 7.2-5). Hughes and Gammon
(1987) and  Miller et al. (1988a) suggest that scores
lying at the extremes of scoring criteria can be  modi-
fied by a plus or minus; a combination of three pluses
or three minuses results in  a  two point increase or
Station No.
Site





Scoring Criteria(b)


1.
2.
3.
4.
5.
(,.
1.
8.
9.
10.
11.
12.

Hetrics(a)
Nuaber of Native Pish Species
Nuaber of Darter or Benthic Species
NuBber of Sunfish or Pool Species
NuBber of Sucker or Long-Lived Species
Nunber of Intolerant Species
X Green Sunfish or Tolerant Individuals
X Omnivores
X Insectivores or Invertivores
X Top Carnivores
Total Nunber of Individuals
X Hybrids or Exotics
X Anomalies
5
m
>67
>67
>67
>67
>67
<10
<20
>45
>5
>67
0
<1
3
~rcr
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

<33
<33
<33
<33
<33
>25
>45
<20
<1
<33
>1
>5
Scorer
GOBI

ents:








Hetric Value Metric Score












IBI Score




(a)

(b)



Karr's original netrics or commonly used substitutes. See text
ties.
Karr's original scoring criteria or commonly
ecoregions.

used


substitutes.

and Table

These may

7.2-1 for other possibili-

require refinement in other

                       Figure 7.2-5.  Data Summary Sheet for Rapid Bioassessment Protocol V.

                                                     7-19

-------
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.
 7.3  RESULTS OF PILOT STUDIES
        IN OHIO AND  OREGON
   Surveys of 109 regional reference sites in five Ohio
ecoregions and of 26 sites on the mainstem Willamette
River, Oregon, were conducted during the summers of
1983 and  1984. The Ohio survey was a cooperative
project between Ohio EPA, EPA Region V, and EPA's
Environmental Research Laboratory at Corvallis
(ERL-C); the Willamette survey was a cooperative
project between DePauw University,  EPA-Region X,
Oregon  Department of Environmental Quality,  U.S.
Fish and Wildlife Service, and ERL-C. The objectives
of the Ohio survey were to evaluate the correspon-
dence between  ecoregions and stream ecosystem
attributes  and to assess regional patterns in attainable
water resource  quality.  The Willamette study was
intended to measure the effects of water resource qual-
ity on the fish community and to examine the useful-
ness of the IBI and IWB in a large western river. The
results of these two surveys have been published else-
where (Larsen et al.  1986; Hughes and Gammon
1987; Ohio EPA 1987b; Whittier et al. 1987; Larsen et
al. 1988)  and only the  fisheries aspects will be sum-
marized here.
                7.3.1 Methods
Ohio
   Minimally-impacted regional reference sites were
selected based on: census and point-source data; maps
of population density, land use, and mining;  and aerial
and ground inspection of the watershed and site (Fig-
ure 7.3-1).
   Ohio EPA collected fish at half the sites two to
three times at  1  month intervals each  summer (1983
and 1984). Rivers were sampled via boat-mounted
electrofishers for 500 meters, headwaters were sam-
pled with  backpack electrofishers for 200 meters,
most streams were sampled with a towed electrofisher
for 300 meters. All captured  fish  were identified to
species, counted, and examined for disease; a subsam-
ple was weighed at the site.  The data  were analyzed
through use of the IBI, IWB, and identification of
regionally characteristic species.
                    ...*..  Regional Reference Sampling Site
                    SSSB Most Typical Areas
                      I   Huron/Erie Lake Plain (HELP)
                      II   Eastern Corn Belt Plains (ECBP)
                     "I  Erie/Ontario Lake Plain (EOLP)
                     IV  Interior Plateau (IP)
                      V  Western Allegheny Plateau (WAP)


Figure 7.3-1. Locations of regional reference sites in Ohio
            (from Whittier et al. 1987).
Willamette River
   Sites were selected along one side of the river
about 2 yd offshore to bracket large point sources and
to space sites approximately  16 km apart (Figure
7.3-2). Each site was 500 meters long and included
slow,  deep water; shallow,  fast, water; and cover.
   Each site was sampled  twice in the summer of
1983 with a boat-mounted  electrofisher.  All fish were
identified to species,  counted, and examined for
anomalies, and a subsample  was  weighed. Data were
analyzed through use of IBI, IWB, and  DCA.

    7.3.2 Results  and Interpretation

   The tolerance and trophic guilds, and origin of
selected species are given in Appendix D. For the
sake of brevity, IBI and IWB calculations are
presented for four selected sites  only.
                                                    7-20

-------
  t
  N
                                              Table  7.3-1
  OREGON
AQUATIC  ECOREGION'S

      WILLAMETTE  VALLEY

      CASCADES

      COAST RANGE
Figure 7.3-2. Locations of sampling sites on the mainstem
          Willamette River, Oregon (taken from Hughes
          and Gammon 1987).


Ohio

   The unmodified IBI metrics and criteria (Karr
et al. 1986) were used to analyze the Ohio data.  Spe-
cies richness metrics were determined as suggested by
Karr et al. 1986 (Figure 7.2-4). Sample data, scoring
criteria, and scores are given in Tables 7.3-1 and 7.3-2
for a site in the Huron Erie Lake Plain (HELP)  and
Western Allegheny Plateau (WAP) ecoregions. The
                                          COLLECTION DATA FOR TWO
                                          OHIO ECOREGION REFERENCE
                                          SITES

                                                            Site
                                                     Species
Grass pickerel
White sucker
Black redhorse
Golden redhorse
Northern hogsucker
Common carp
Blacknose  dace
Creek chub
Golden shiner
Redfin shiner
Silver shiner
Rosyface shiner
Striped shiner
Sand shiner
Mimic shiner
Silverjaw  shiner
Bluntnose  minnow
Fathead minnow
Central stoneroller
Yellow bullhead
Black bullhead
Tadpole madtorn
Brindled madtom
Blackstripe topminnow
White crappie
Green sunfish
Bluegill
Rock bass
Longear sunfish
Smallmouth bass
Largemouth bass
Orangespotted sunfish
Blackside  darter
Logperch
Johnny darter
Greenside  darter
Banded darter
Rainbow darter
Fantail darter
                                                       WAP
                                                                           21
                                                                            2
                                                                           24
                                                                           26

                                                                            3
                                                                           94
                                                                            6
                                                                           41
                                                                          443
                                                                          264
                                                                            1
                                                                           93
                                                                          101

                                                                          559
                                                                           16
                                                                            3
                                                                           11
  3
  3
 40
 56
 25
 60
113
         HELP

          19
          24
                                                                  26

                                                                   2
                                                                  42
                                                                  22
                                                                   8
                                                                   3

                                                                  23
                                                                  13
                                                                   2

                                                                  66
                                                                   6
                                                                 156
                                                                   1
                                                                                      2
                                                                                      2
                           fish communities of the Ohio regional reference sites
                           showed distinct ecoregional differences between the
                           Western Allegheny Plateau and the Huron Erie Lake
                           Plain ecoregions. Differences among the three transi-
                           tional ecoregions were less obvious. This was true
                           whether examining patterns in IBI scores (Fig-
                           ure 7.3-3) or dominant species (Figure 7.3-4).
                                           7-21

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          TABLE 7.3-2
           SCORING CRITERIA AND IBI  AND IWB SCORES
          FOR TWO OHIO  ECOREGION REFERENCE SITES
                Metric  (Criteria)
                                                            Value(Score)
                                              WAP
Total Number of Species  (<9=1,  9-18=3, >18=5)
Number  of  Darter Species  (<2=1,  2-5=3, >5=5)
Number  of  Sunfish Species  (<2=1,  2-4=3, >4=5)
Number  of  Sucker Species  (<2=1,  2-4=3, >4=5)
Number  of  Intolerant Species  (<4=1,  4-8=3, >8-5)
% Green Sunfish   (>20=1,  5-20=3,  <5=5)
% Omnivores (>45=1, 20-45=3,  <20=5)
% Insectivorous Minnows  (<20=1,  20-45=3, >45=5)
% Top Carnivores (<1=1,  1-5=3,  >5=5)
Number  of  Individuals  (<200=1,  200-800=3, >800=5)
% Hybrids  (>1=1, 0-1=3,  0=5)
% Diseased (>5=1, 2-5=3, <2=5)

Total IBI  Score

IWB Score

Integrity
                                               25(5)
                                                7(5)
                                                2(3)
                                                4(3)
                                                8(3)
                                                0(5)
                                                5(5)
                                               42(3)
                                                1(3)
                                            2,012(5)
                                                0(5)
                                                1(5)

                                                  50

                                                  10

                                                Good
 HELP

 17(3)
  0(1)
  4(3)
  1(1)
  0(1)
 37(1)
  9(5)
 15(1)
  5(3)
417(3)
  0(5)
  4(3)

    30
  Poor
          UJ
          cc
          O
          o
          CO
          5
              r
              •o
              §
   50-
                46-
 F  •
•£42-
                38-
                34-
              o
              o
              a
              L30

                       HELP   ECBP  EOLP    IP

                             'ECOREGION
                                   WAP
      Figure 7.3-3. Index of biotic integrity scores by Ohio ecoregion (from Whittier et al. 1987).
                Vertical lines represent ranges; horizontal lines represent 10th, 25th, 75th, and
                90th percentiles; open circles are medians.
                                    7-22

-------
                1   bluntnose minnow
                2   creek chub
                3   green sunf ish
                4   blackstripetopminnow
                5   fathead minnow
                6   yellow bullhead
                7   johnny darter
                8   white sucker
                9   rockbass
               10   rosefin shiner
               11   greenside darter
               12   bluegill
               13   mottled sculpin
               14   common shiner
               15   rainbow darter
               16   fantail darter
               17   spotfin shiner
               18   longear sunf ish
               19   sand shiner
               20   golden redhorse
               21   emerald shiner
               22   central stoneroller
               23   striped shiner
  100
c
ro
i  60
   20
            llllll
       1  2  3  4 5  6  7  8 9 10 11 12 13 14 15 16 17 1819 20 21 22 23
                          fish species

                   Huron/Erie Lake Plain
80
0)
w
C
2 60
V
0
g 40
V
fll
a
20

















Im
1
ill ll
80
a>

c
3 60

O
v 1°
W
a>
Q.
20











1
I 1
III i i ill i Illlll 	 II
1  23  456  7  8  9 10 11 12 13 14 15 16 17 18 19 20 21 22 23
                   fish species
           Eastern Corn Belt Plains
       1  2  3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 1819 20 21 22 23
                          fish species

                  Erie/Ontario Lake Plain

80

-------
Willamette River
   Five of the original 12 metrics that Karr et al.
(1986) found appropriate  for midwestern streams were
inappropriate for a large  western river. These five
metrics  were modified by making the following substi-
tutions:  (1) The number of sculpin species replaced
the number of darter species as suggested by Karr et
al. (1986); (2)  The number of native minnow species
replaced the number of sunfish  species. In cool and
warmwater western streams, the introduced sunfish
increase and native minnows decline when habitat
structure deteriorates (Minckley 1973; Moyle 1976);
(3) Percent common carp replaced percent green sun-
fish. No green sunfish were collected in the Wil-
lamette, the  other relatively tolerant species were
either rarely captured or  dominant, and common carp
increased as the physicochemical habitat deteriorated;
(4) Percent catchable salmonids (longer than 20 cm)
replaced percent piscivores. The dominant piscivore in
the Willamette is a relatively tolerant species and is
not indicative of integrity; most Willamette salmonids
(juvenile whitefish and anadromous salmon) are not
piscivorous in  freshwater rivers; and (5) Percent
introduced individuals  replaced  percent hybrids.
Hybrids are too rare in the Willamette to make this a
useful metric for discriminating degradation. Percent
introduced individuals do increase with degradation
(Moyle and Nichols 1973; Holden and. Stalnaker 1975;
Leidy and Fiedler.  1985) and represent a loss of the
segregation existing before midwestern species were
brought to western waters.
   Scoring criteria were based on the Willamette data
due to the lack of sufficient quantitative  historical
data; no adjustments were required for stream size
because  it changed little. Criteria were based on
characteristics of the fish assemblages at the
minimally impacted sites of the upper mainstem
(Table 7.3-3). Like the metrics, criteria were adjusted
in the following ways to reflect western conditions
(Table 7.3-4): (1) Criteria for common carp were
reduced  because carp are much less common in
Oregon than the green sunfish is  in the Midwest;
(2) Omnivore criteria were increased because a domi-
nant species (largescale sucker) is an omnivore;
(3) The catchable salmonids criteria were increased
because  salmonids are more common than midwestern
piscivores; (4) Percent introduced individuals had
   TABLE  7.3-3   COLLECTION  DATA (NUMBER  OF INDIVIDUALS) FOR  TWO  SITES  ON THE
                    WILLAMETTE  RIVER,  OREGON
                                                                                        River
                                                                                     Kilometer
           Species
    Mountain  whitefish
    Chinook salmon
    Longnose  dace
    Speckled  dace
    Northern  squawfish
    Common carp
    Largescale  sucker
    Mountain  sucker
    Largemouth  bass
    Yellow perch
    Prickly sculpin
    Reticulate  sculpin
    Torrent sculpin
    Paiute sculpin
                                270

                                25
                                 4
                                 1
                                 3
                                 2

                                44
                                 6
                                 2
                                 3
                                 5
31
 2
 7
10

 2
 1
 1
                                                 7-24

-------
            TABLE 7.3-4  SCORING  CRITERIA AND IBI  AND  IWB  SCORES  FOR
                           TWO SITES ON THE  WILLAMETTE RIVER, OREGON
                     Metric  (Criteria)
  Number of Native Species  (<5=1,  5-9=3, >9=5)
  Number of Sculpin Species  (<2=1,  2=3,  >2=5)
  Number of Native Minnow Species  (<3=1, 3-5=3,  >5=5)
  Number of Sucker Species  (0=1,  1=3,  >1=5)
  Number of Intolerant Species  (0=1,  1-2=3,  >2=5)
  % Common  Carp  (>9%=1,  l-9%=3,  50%=1, 25-502=3,  <25%=5)
  % Insectivores  (<20%=1, 20-40%=3, >40%=5)
  % Catchable Salmonids  (0%=1,  l-9%=3,  >9%=5)
  Number of Individuals/km  (<50=1,  50-99=3,  >99=5)
  % Introduced  (>9X=1, l-9%=3,  <1%=5)
  % Anomalies (>5%=1,  2-5%=3,  <2%=5)

  Total IBI Score

  IWB Score

  Integrity
                                                                        Value(Score)
                                                                      River Kilometer
                                        270
31
higher criteria than expected for percent hybrids; and
(5) Percent anomalies criteria were higher because,
where they occurred, they were above the 1  percent
maximum as referenced in Karr et al. (1986).
   A detrended correspondence analysis (DCA)
showed four distinct clusters among the 26 sites cor-
responding to upper river, middle  river, Newberg
pool, and Portland metropolitan fish  assemblages
(Figure 7.3-5). These differences and the longitudinal
                                                    4(1)
                                                    1(1)
                                                    KD
                                                    1(3)
                                                    1(3)
                                                   28(1)
                                                   68(1)
                                                   16(1)
                                                    0(1)
                                                   25(1)
                                                   40(1)
                                                    8(1)

                                                       16

                                                      4.7

                                                  Very Poor
                 trends shown by the IBI (Figure 7.3-6) corresponded
                 to declining physical and chemical habitat quality and
                 increased nonpoint-source pollution.
                 (l)Karr et al. (1986); (2)Hughes and Gammon (1987); (3)Miller et
                 al., (1988b In Review); (4)Miller et al. (1988a); (5)Ohio EPA
                 (1987b); (6)Metric suggested by Moyle or Hughes as a provisional
                 replacement metric in small western salmonid streams; (7)Leonard
                 and Orth (1986); (8) Steedman (1988).
Upper
River
                             Portlond
                             Metro
                180-


                160-


                140


                120-


              c^ 100
              c/»

              •5  80-


                 60-


                 40-


                 20
                       20  40  60  80  100 120  140  16O  18O  200 220  240  280
                                           Axis 1

           Figure 7.3-5.  Patterns in mainstem Willamette River fish assemblages as revealed by
                      detrended correspondence analysis (taken from Hughes and Gammon 1987).
  Middle
  River
                                           7-25

-------
     60
     50
     40
=    30
     20
     10
              IBI
              NO.3 x 100
       300
                                     200
100
                                                 River Kilometer
              Figure 7.3-6.  Longitudinal trends in Index of Biotic Integrity and nitrate in the
                           Willamette River (modified from Hughes and Gammon) 1987.
                                                7-26

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                        8.  INTEGRATION OF  HABITAT,
                WATER QUALITY, AND  BIOSURVEY DATA
   The overall assessment of ecological condition first
focuses on the evaluation of habitat quality, then ana-
lyzes the biological components of the system in light
of these data. Habitat, as the principal determinant of
biological potential,  sets the context for interpreting
biosurvey results arid can be used as a general predic-
tor of biological condition.  Routine water chemistry
can also help to characterize certain impacts.
   In Rapid Bioassessment Protocols (RBPs) I and IV,
the habitat evaluation carries considerable weight in
the final'assessment because minimal  effort is
expended on the collection  and analysis of biological
data. In RBPs II, III, and V, however,  the biological
evaluations are more rigorous and appropriately take
precedence. The habitat assessment plays a supporting
role within these protocols. It is used to identify obvi-
ous constraints on the attainable potential of the site,
help in the selection of appropriate sampling stations,
and provide basic information for interpreting biosur-
vey results.
       8.1 THE RELATIONSHIP
  BETWEEN HABITAT  QUALITY
  AND BIOLOGICAL  CONDITION
   The attainable biological potential of a site is
primarily determined by the quality of the habitat at
that site. The relationship between habitat quality and
biological condition can be envisioned as a sigmoid
curve (Figure 8.1-1) with community response varying
with habitat quality. In the upper segment of the
curve, good quality habitat (supporting or excellent)
will support high quality communities, and responses
to minor alterations in habitat will be only subtle and
of little consequence. As habitat quality continues to
decline, however, discernible biological impairment
results, and, in the absence of confounding water
quality effects, the relationship is roughly linear.
            100
                                                                                  90     100
                                     Habitat Quality (% of Reference)
                    Figure 8.1-1. The relationship between habitat and biological condition.
                                                 8-1

-------
    In areas of severe habitat degradation, predicting
the degree of biological impairment becomes more
difficult. Community structure is less dependent on
habitat diversity, which is effectively simplified by
degradation, and more dependent on the opportunistic
colonization strategies of a relatively few tolerant spe-
cies. These opportunists are adapted to environmental
conditions that are unfavorable to most other species
and, in the absence of competition, thrive (or at least
survive) in these marginal conditions. Therefore, bio-
logical measures, particularly those used  in the RBPs,
are relatively insensitive to habitat  variations  in this
range, and a nonsupporting  characterization may cor-
respond  to either a moderately or severely impaired
biological  condition, depending on the specific site.
    When habitat and biological data are systematically
collected together, empirical  relationships can be
quantified  and subsequently used for screening impact
sites, scoping field activities, and discriminating water
quality impacts from habitat degradation. With the
acquisition of a multiple-site database,  confidence
bounds can be established for the habitat/indigenous
community relationship.
    A theoretical relationship of habitat quality and
                                    biological condition as affected by water quality prob-
                                    lems (organic or toxicant loadings) also can be
                                    hypothesized (Figure 8.1-2). Curve II in Figure 8.1-2
                                    indicates the general relationship of biological condi-
                                    tion to habitat  quality in the absence of water quality
                                    effects. Curve  II may, in fact, resemble a sigmoid
                                    curve as illustrated by  Figure  8.1-1. Curve III repre-
                                    sents a situation where organic pollution or toxicants
                                    will adversely  affect biological condition regardless of
                                    the quality of the habitat.
                                       In areas of good or excellent habitat, biological
                                    communities will reflect degraded conditions when
                                    water quality effects are present.  However, as habitat
                                    degrades to a poor condition in the presence of water
                                    quality problems, response of the communities may be
                                    less dramatic because the community is composed of
                                    tolerant and generally opportunistic species. Curve I is
                                    representative of a situation indicative of nutrient
                                    enrichment, which will artificially sustain a more
                                    diverse  fauna than dictated by the habitat quality.
                                    However,  at some point along the curve as habitat
                                    degradation proceeds, nutrient enrichment will no
                                    longer support a diverse community, and  a drastic
                                    decrease in biological condition will result.
                o>
                o
                o
o
O
75
o
w
o
o
3
                                                      Habitat Quality
                                                          Decreasing

          Figure 8.1-2.  Relationship of habitat quality and biological condition in the context of water quality.
                                                       8-2

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          8.2 BIOASSESSMENT
                TECHNIQUE
   As described in Chapters 6 and 7, the biological
assessment involves an integrated analysis of both
functional and structural components of the aquatic
communities. These functional and structural compo-
nents are evaluated through  the use of eight metrics
for benthic RBPs H and III  and  12  metrics for fish
RBP V. The range of pollution sensitivity exhibited by
each metric differs among metrics (Figures 8.2-1 and
8.2-2); some are sensitive across a  broad range of bio-
logical conditions, others only to part of the range.
Sensitivity  of metrics  may also vary depending on
whether organic or toxicant  impacts are being evalu-
ated  (Figure 8.2-1). The considerable overlap in the
ranges  of sensitivity helps reinforce final conclusions
        regarding biological condition, while metrics that are
        better able  to differentiate responses at the extremes
        of the range of impairment enable a more complete
        bioassessment.  The integrated analysis approach thus
        allows a broader assessment of condition than an anal-
        ysis using any single metric.  However, information
        from individual metrics will be useful in enhancing
        overall data interpretation.
           Certain  metrics are designed  to be better estima-
        tors of either organic or toxicant effects.  For instance
        in the benthic protocols, the Hilsenhoff Biotic Index
        (HBI) utilizes a tolerance classification scheme that is
        based on organic  pollution  effects, while Functional
        Group representation can be altered by either organics
        or toxicants (Figure 8.2-1).  Although Scrapers and  •
        Filterers are affected by toxicants to a certain extent,
        their ratio can best be  used to assess'organic enrich-
        ment (Cummins 1987, personal communication). As
        discussed in Chapter 6, a reduction in the value
                      Organics
                               Metrics
                       Taxa Richness
                       HBI
                       FFG* Scrapers/Filterers
                       EPT Abund./Chiron.Abund.
                       % Contribution (dom.taxon)
                       EPT
                       Communily Similarity Index
                    £ [ FFG •» Shredders/Total
                    o
                                                           Biological Condition
 Non-
Impaired
Severely
Impaired
                            I	1

                           —I
                      Toxicants
                               Metrics
                        Taxa Richness
                        HBI
                        FFG* Scrapers/Filterers
                        EPT Abund./Chiron.Abund.
                        % Contribution (dom.taxon)
                        EPT
                        Community Similarity Index
                      [ FFG •» Shredders/Total
                                                           Biological Condition
Non-
Impaired
Severely
Impaired















            Figure 8.2-1.  Range of sensitivities of Rapid Bioassessment Protocol II and III benthic metrics
                        in assessing biological condition in response to organics and toxicants.
                                                     8-3

-------
obtained for Scrapers/Filterers can be indicative of
either a reduction in the quality of the periphyton as a
food  source and/or an increase in the suspended
FPOM. Filterers are also affected  by FPOM contami-
nated by toxicants.
   The relative abundance  of Shredders in the benthic
community is a good indicator of toxicant problems
because of the sensitivity of the Shredder community
to toxic conditions (Cummins 1987, personal commu-
nication). Vegetation sprayed with  pesticides eventually
becomes a CPOM food source for Shredders. In  suffi-
cient  concentrations, toxicants bound to CPOM may
affect Shredders directly through ingestion,  as well as
indirectly by killing attached microbes that serve  as a
nutrition base for Shredders. The ratio of the abun-
dances of EPT taxa and chironomids may also  func-
tion as a toxicant  indicator, since some midge species
such  as Cricotopus sp.  become abundant in areas
affected by metals (Winner et al. 1980; Mount et al.
1986).
   The 12 IBI metrics used in fish Protocol  V also
represent  differing sensitivities  (Figure 8.2-2). For
example, municipal  effluents typically affect  total
abundance and trophic structure (Karr et al.  1986),
while unusually low total abundance generally indi-
cates  a toxicant effect. However, some nutrient-
deficient environments support a limited number of
individuals, and an increase in abundance may indi-
cate 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 Harbour  1983;  Ohio EPA
1987b) and are good indicators of habitat degradation.
   For the benthic and fish biosurveys 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 function of a
range of scores and is weighted  in terms of its contri-
bution to the total habitat quality. The scores assigned
to the benthic and fish metrics are based on computed
values of the  metrics  and a station comparison,  where
the regional or stream reference station serves as the
highest attainment criterion. Comparison  of the  total
score computed for the metrics or parameters with
that of the reference station provides a judgment as to
impairment of biological condition.
   Effects indicated by the aquatic community need to
be evaluated in the context of habitat  quality. A poor
habitat in terms of riparian vegetation, bank stability,
stream substrate,  etc., would not be conducive to sup-
porting a well-balanced community structure.  The
attainment of a higher quality biological  condition
may be prohibited by the constraints of habitat quality.

  Biological Condition
Metrics
Species
Darters
Sunfishes
Suckers
Intolerants
% Green Sunfish
% Omnivores
% Insectivorous Cyprinids
% Piscivores
Number
% Hybrids
% Diseased
Non- Severely
Impaired Impaired





u> < » . .I i





                     Figure 8.'2-2.  Range of sensitivities of Rapid Bioassessment Protocol V fish
                                  metrics in assessing biological condition (from Karr et al. 1986).
                                                      8-4

-------
         8.3  AN INTEGRATED
      ASSESSMENT APPROACH
   The initial focus of a bioassessment should be on
habitat quality. Based on a regional  reference, the hab-
itat at an impacted site may be equal to or less than
the desired quality for that particular system. As dis-
cussed in Section  1.4, if the habitat  at the impact site
and reference are  equal, then a direct comparison of
biological condition can be made. If the habitat at the
impact site is lower in quality than the reference, the
habitat potential should be evaluated as a first step. A
site-specific control may be more appropriate than a
regional reference for an assessment of an impact site.
Once a determination of the appropriate reference site
type is made, possible outcomes of the bioassessment
are: (1) no biological effects; (2) effects due to habi-
tat degradation; (3) effects due to water quality; or
(4) effects due to a combination  of water quality and
habitat degradation.  Once habitat problems are identi-
fied, in most cases,  separating the cause  of impair-
ment from water quality problems is difficult. The
following decision matrix illustrates the approach to
assessing biological  effects.

Evaluation of Habitat at a Site-Specific
Control Relative to that at a Regional
Reference (Figure 8.3-1)
   Selection of an appropriate station of  comparison
for evaluation of biological impact begins with an
evaluation of habitat at the potential control  station.
                    HC+RHA=HR
                           (II)
               HC + RHA

-------
This comparison assumes that a regional reference
database is available for the particular site being stud-
ied. Reference data used for comparison may be
obtained from a single reference site. However, a
reference database  derived from numerous sites is
much preferred and strongly recommended.
    Scenario I depicts  the situation where the habitat
quality  at the control station (HC) is equivalent to that
at the regional reference (HR). If the control station
habitat  is degraded relative to  that at the regional
reference, it becomes  necessary to consider  the effect
that reversible habitat  alterations (RHA) may have on
habitat  quality (Scenarios II and III). Reversible habi-
tat  alterations are those habitat parameters that can
potentially be altered by remedial action (i.e., bank
stability, bank vegetation, streamside cover,  and some-
times embeddedness).

Evaluation of Water Quality  Effects
(Figure  8.3-2)
    A determination may be made that the habitat
quality  at the site-specific control station (C) is
equivalent to that at the reference (R) (Scenario I). In
this case, a biological assessment can be used to
evaluate potential water quality effects at C  (Fig-
ure 8.3-2).

(1)  If impairment  is not detected in a comparison of
    biological condition at the site-specific control
    station (BC) to biological  condition at the refer-
    ence (BR), then C should be included in the R
    database  and either C or R may be used as a
    reference for biological assessment at the impact
    site (I). The site-specific control would  be the
    best indicator of a site-specific situation. In addi-
    tion, C would  be more appropriate for  use in
    determining water quality  effects of point-source
    pollutants since it would be located on the same
    waterbody (or  a nearby waterbody) and would
    integrate  all other background sources of impair-
    ment other than the point source being  evaluated.
    This allows segregation of effects of a particular
    point source. The reference would be more
    appropriate in  an assessment of nonpoint-source
    effects since it is  virtually  impossible to find a
    nearby  site-specific control which would not  be
    impacted by the impact sources being studied. If
    R is based on  an  extensive database, then use of
    R as a  reference would provide  a better estimate
    of acceptable variation in  a data set. Confidence
    intervals  could be  derived and used to put bounds
    on  the data from  C and/or the  impact site (I).

(2)  If biological impairment is detected at C relative
    to R, the impairment may be attributed  to water
    quality  effects. The use designation at C is proba-
    bly appropriate, but R should be used as the bio-
    assessment reference because of impairment at C.

Evaluation of Biological Impairment  Due
to Reversible Habitat Alterations
(Figure 8.3-3)
   If the habitat quality at C is  degraded relative to
that at R, but habitat quality could potentially  be
improved by reversing those degraded  habitat parame-
ters which  are reversible (Scenario II), biological
assessment at C will indicate whether  C, R, or an
alternative  control site (C*) should be  used as  a
bioassessment reference for the  impact site  (I)
(Figure 8.3-3).

(3)  When  BC is  equal to BR, the use designation at
    C may be considered appropriate, and C should
    be used as the bioassessment reference.  This is a
    potential situation since reversible habitat param-
    eters are mainly tertiary characteristics and
    should have the least effect  on the biological
    community. However,  in this situation benthic
    RBP III or fish RBP V should be utilized  as a
    minimum. A more rigorous biological analysis
    (e.g., quantitative sampling) may be warranted to
    ensure that the approach is  sufficiently sensitive
    to detect impairment.

(4)  In situations  where BC is less than BR, impair-
    ment may be due to either  reversible habitat alter-
    ations, water quality effects, or a  combination of
    the two. Selection of a bioassessment reference is
    dependent on the  purpose of the assessment and
    the suspected source of impairment.

(5)  If point source effects are being assessed,  a habi-
    tat independent approach (e.g., toxicological test-
    ing, sampling with artificial substrates) may be
    warranted, using R as a reference. C could be
    considered an impact site.

(6)  It may be appropriate to continue  the rapid bioas-
    sessment (and habitat evaluation) approach using
    R as the reference because  of impairment  at C.
    Assuming that impairment at C indicates impair-
    ment also at  I, the degree of impairment at I can
    be assessed relative to C. An a priori knowledge
    of potential water quality problems from an exist-
    ing database  would enhance interpretation  of find-
    ings in this case.

(7)  Another alternative would be to eliminate  the
    confounding  effects of the reversible habitat alter-
    ations  by selecting another  site-specific control
    station (C*),  which, if possible, would then be
    evaluated relative  to R (Figure 8.3-1).
                                                      8-6

-------
                                      HC = HR
                                         (I)
                   BC = BR
               HC = HR; BC = BR
                Include "C" in
                "R"  database.
                      (1)
       BC

-------
           HC + RHA =  HR
               BC = BR
           Use  designation
           is appropriate.
                  (3)
             Use  "C"  as
            bioassessment
             reference.
Continue RBP—
habitat and
biosurvey
approach .
(6)
i
r


« — 1


                                       HC + RHA = Hft
                                           BC  < BR
                                      Impairment at "C"
                                    due to revs hab  alts
                                      •(-/or WQ effects.
                                         Use habitat
                                         independent
                                          approach
                                       (toxicological.
                                      art.  substrates)
                                             (5)
                                                               Use  "R" as
                                                              bioassessment
                                                               reference.
                                                             Consider "C" as
                                                             an impact site.
   Use "C" as
   control if
habitat quality
    is best
   attainable
Use ecoregional
  database as
 bioassessment
   reference.
Consider  "C" an
  impact site.
Evaluate
habitat
 at new
control
    Figure 8.3-3. Evaluation of biological impairment due to reversible habitat alterations
                (RHAs).  (Numbers in parentheses refer to points of discussion in text.)
                                        8-8

-------
Evaluation of an Alternative Site-Specific
Control Station  (Figure 8.3-4)

   If the habitat quality at C is degraded relative to
that at R, and reversible habitat alterations  do not
account for all of the habitat differences, then it is
necessary to select an alternative site-specific control
station (C*), if one  is available (Figure 8.3-4).
(8) If a more appropriate control is located (where
    HC* = HR) then use C* as the  reference and pro-
    ceed with the bioassessment.

(9) If HC* is also degraded relative to HR, and it
    appears that a better quality site-specific control
    is not  available, then biological condition  should
    be evaluated at  C* relative to R.

(10) Where reversible habitat alterations account for
    all differences in habitat quality between C* and
    R, then use C*  as  the reference and proceed with
    the bioassessment.

(11) In the unlikely situation where  the degraded habi-
    tat (reversible and/or irreversible parameters) at
    C* is  not limiting to biological condition, and
    BC* = BR, then either C* or R would be  an
    appropriate reference.

(12) If biological impairment is detected at C*  the
    effects may be attributable to either degraded hab-
    itat  (reversible and/or irreversible parameters)
    and/or water quality effects. Three possible alter-
    natives should be considered: (a) A Use Attain-
    ability Analysis (UAA) is  needed to determine the
    appropriate use  classification of the system. In
    this case, the system as represented by C  or C* is
    aberrant to R. A UAA will  be  needed to  redefine
    R* or a subset  of R, for interpretation of an
    appropriate bioassessment. (b) C* would be used
    as a reference for bioassessment because it
    represents the best attainable condition for that
    system. However, interpretation of effects, would
    be in the context of a control that does not meet
    the criteria of the region,  (c) A prediction of the
    expected biological  condition can be made from
    an extrapolation of  the regression line formed
    from the  reference database and the best potential
    habitat quality at C*.
Bioassessment Using a Site-Specific
Control Station  (Figure 8.3-5)
   Once the decision is made to use a site-specific
control  (C or C*) then  evaluation of the impact site
relative to C (or C*) proceeds as in Figure 8.3-5.  As
indicated  in the previous flowcharts, C is used when it
is biologically representative of the region or  is con-
sidered to represent the best attainable condition. A
matrix of conclusions from the potential scenarios is
presented in Table 8.3-1. The general  bioassessment
approaches  are as follows:

(13)  If HC = HI, then bioassessment for the purpose of
     detecting water quality effects at  I (impact site)
     would proceed  similarly to the evaluation of BC
     relative to BR (Figure 8.3-2).

(14)  Where reversible habitat alterations account for
     all habitat differences between C  and I, then
     bioassessment would proceed as  in Figure 8.3-3.

(15)  If habitat degradation is due to reversible and/or
     irreversible alterations, then bioassessment would
     proceed as in Figure  8.3-4.

Bioassessment Using a Regional Reference
(Figure  8.3-6)
   In situations where  R is to be used as a reference,
reference data could be obtained either from a single
reference site or a regional database made up of
numerous sites, and evaluation of the  impact site
would proceed as in Figure 8.3-6. As data are
accumulated and processed, regional  databases will
provide refinement to the  criteria and enhance bio-
assessments. A matrix  of conclusions that would
result from the possible scenarios is presented in
Table 8.3-1.

(16) If HR = HI, then the  approach in assessing poten-
     tial water quality effects at I would be similar to
     that  followed in evaluating BC relative to BR
     (Figure 8.3-2).

(17)  Where reversible habitat alterations account for
     all habitat differences between I and R, then
     bioassessment would proceed as  in Figure 8.3-3.

(18)  Where habitat degradation may be caused by
     reversible and/or irreversible alterations, then
     bioassessment would proceed according to Fig-
     ure 8.3-4.
                                                     8-9

-------
HC
 RHA <
(III)
HR
                                   Select  C*
   HC*  = HR
       IB)
HC*
+ RHA
.19)
< HR
                                                   HC* + RHA
                                                         (10)
                                                                         HR
    Go to
      (I)
  BCX = BR
  (unlikely
 scenario)
    (11)
HC + RHA < HR;
BC = BR
Habitat not
limiting;
No bioimpact
at "CH"
*


Use "C*" or
'R'as
reference for
bioassessment








^





(a




HC* + HHA < HR BC* < BR
Bioimpact at "C*n.
Effects due to degraded
habitat +/or WQ
(UAA is eventually
needed)






^ ,


Perform UAA.
Redef ine'Rx"
or subset of'R"







(b)
Consider "CK"
best attainable
condition. Use
°C*° as
reference for
bioassessment.




r
Icl
Predict
biological
condition using
known habitat
qua'lity and
reference
database.
        Figure 8.3-4. Evaluation of an alternative site-specific control station (C*).
                    (Numbers in parentheses refer to points of discussion in text.)
                                       8-10

-------
                            (Using  "C"
                            [or  "C*"]
                               as a
                            reference
                            station)
Figure 8.3-5.  Bioassessment using a site-specific control station.
             (Numbers in parentheses refer to points of discussion in text.)

-------
                               TABLE 8.3-1  BIOASSESSMENT CONCLUSIONS  RELATIVE  TO USE OF  A SITE-SPECIFIC CONTROL OR  REGIONAL  REFERENCE
                                                                                      HC  *  RHA
                                                                                                                                 HC t  RHA  <  HR
oo
fo
BC = BR BC
No bioimpairment at C
or I ; I is a candi-
n date for inclusion in
th* R database .
u
CO
M
R Bioiopaiment at I
0 due to WQ effects .
X
M
0
U
Q

o I; reversible habitat
I alterations are
w present but not
n liaiting.
u
X
* Bioiapairnent at I
< due to degraded
« habitat (reversible
t parameters) and/or
M WQ.|B)
M CO
X
u
n
(a) Use attainability analysis
(b) To differentiate water qual
< BR BC = BR

or I; use designation
appropriate. X is a
candidate for inclu-
sion in R database.
Reversible habitat
alterations arc pre-
sent but not limiting
Bioimpairment at I .
due to WQ effects
sible habitat
alterations.
'Unlikely scena'rio
Bioinpairaent at I
due to degraded
paraneters) and/or WQ
effects.
eventually needed.
ity effects from habitat effects, a
BC < BR(a) BC = BR
Unlikely scenario
Unlikely scenario
Unlikely scenario
Bioimpiiraent at I
due Co degraded
habitat (rever-
sible w/respect
to C, and both
reversible and
irreversible para-
aeters w/respect
to R) and/or WQ
effects.
fairly extensive database on water quality
BC < BRIa)
Bioimpairment at C
and I due to
(reversible and/or
meters) and/or WQ
effects. TbT
Both C and I
impaired relative
to R due to
(reversible and/or
irreversible para-
meters) and/or WQ
effects. ' Addi-
ment at I due to
WQ.
Bioimpairment at C
and I due to
degraded habitat
(reversible and/or
irreversible para-
meters) and/or WQ
effects.'01
Both C and I
inoaired relative
to R due to
degraded habitat
(reversible and/or
irreversible
parameters ) and/or
WQ effects. 1DI
Additional
due to degraded
parameters I .
parameters is
                        (0
                            necessary.  This  information,  if  it  exists,  should  be  available Cor prior review.   Thus, an agency would b« aware of a
                            potential water quality  problea prior  to   biological assessment,  which would aid in the determination of course  and effects
                            relative to habitat  constraints or  water  quality  probleas.
                            Unlikely, but  nay  occur  in  response  to organic  enrichment.

-------
TABLE  8.3-1  (Cont. )
                  HR
                                                  HC  +  RHA  < HR
                     BC <  8RIa I
                                                               BC < BRIa)
M
m
n
u
03
H
a
u
m
M
n
M
K
ra
M
O)
tt
m
NO DLO impairment at
I, but reversible
and/or irreversible
habitat alterations
present .
Bioimpaiment at I,
duo to degraded
habitat ( reversible
and/or irreversible
parameters) and/or WQ
effects. '

Include I in R
database .
due to WQ effects.


para no tors) and/or WQ (reversible and/or

WQ effects. ( '
Biological impaitxent Unlikely scenario Unlikely scenario
at C, but not I .
Both C and I impaired C impaired relative to Bioimpairnent at I
relative to R due to Rduetodegraded dueto WQ . Habitat
parameters) and/or WQ
effects. Bioimpair-
nent at I due to WQ
effects .
nioiapairment ac I,
and C due to
degraded habitat
( reversible and/or
irreversible
parameters ) and/or
WQ effects. ' '
Both C and I
to R due to
degraded habitat
(reversible and/or
parameters ) and/or
WQ effects .
Addi tional bio-
to degraded habitat
( reversible and/or
itat parameters)
and/or WQ . '
Unlikely scenario
C iap aired relative
to R due to
degraded habitat
( reversible and/or
•eters ) and/or WQ
effects.' ' Bio-
                                                         to degraded habitat
                                                         (reversible para-
                                                         Be ters) and/or WQ.

-------
                                                                              TABLE  8.3-1   ICont.)
                                              HR
                                                                                      HC  +  RHA  =  HR
                                                                                                                                 HC *  RHA <  HR
00
BC = BR

alterations present,
but not Uniting.
Bioimpairoent at I
habitat alterations
and/or WQ •f«»cts.lcl


BC < BR

habitat parameters
and/or WQ effects .
due to WQ.
C impaired relative
effects . Bioimpair-
ment at I due to
(reversible para-
meters) and/or WQ . '

BC = BR BC < BR(a) BC = BR
Unlikely scenario Un likely scenario

Dioimpairment at C and Bioimpairaent at X
reversible habitat habitat altera-
alterations and/or WQ . tions and/or WQ .
1 imi ting at C .
Unlikely scenario Unlike ly scenario
BC < BRtal
Unlikely scenario

Bioiapairaent at C
due to degraded
habitat (reversible
for C , reve rs ible
for I) and/or WQ .
Unlikely scenario
                       I.  but reversible
                       and/or irreversible
                       habitat alterations
                       a re present.
                       Bioiapaicment at I
                       due to degraded
                       habitat  (reversible
                       ar.H/or irreversible
                       paranecersi and/or MQ
                       effects.101
due to degraded
habitat '{reversible
and/or irreversible
parameters)  and/or
WQ.     Bioimpairment
at C due to WQ.

C impaired relative
to R due to WQ
effects.  Bioimpair-
aent «*t I due to
degraded haoitat
(r»>v*rsible and/or
irreversible para-
meters) and/or WQ.
Bioinpai raent at C
relative to R due to
reversible habitat
alterations and/or WQ.
Bioiapairment at I due
to degraded habitat
(reversible and/or
irreversible para-
meters) and/or WQ
effects.top
Bioinpairaent at
I due to degraded
habitat and/or WQ.
Habitat at C ttot
limiting.
Bioimpairment at C
and I relative to R
due to degraded
habitat (irrever-
sible and rever-
sible parameters)
and/or WQ.

-------
                              R"
(Using
   as  a
reference
 station)
Figure 8.3-6.  Bioassessment using a regional reference.
             (Numbers in parentheses refer to points of
             discussion in text.)
                         8-15

-------
              8.4  CASE STUDY
                                                                   HI+RHA = HR — HI + RHA VS
   Using the data from the North Carolina DEM pilot
study (discussed in Section 6.4), an  integrated assess-
ment can be performed using the decision matrix
approach described in Section 8.3. This case study is
presented in a step-by-step fashion to illustrate the
concepts of the decision matrix. Only data from Sta-
tions 3 and 4 are compared to Stations 1  (site-specific
control)  and R (regional reference).

1.  The habitat quality of the site-specific  control is
   first compared to that of the regional reference.

          HC = HR «- HC VS HR -* HC

-------
         TABLE 8.4-1  SUMMARY OF  HABITAT ASSESSMENT  SCORING  FOR ARARAT AND MITCHELL RIVERS BENTHIC  CASE  STUDY DATA
00

Habitat Category/Parameter
Primary — Substrate and Ins t ream Cover
1. Bottom substrate and available cover
2. Embeddedness^ '
3. Flow/velocity
Secondary — Channel Morphology
4. Channel alteration
5. Bottom scouring and deposition
6. Pool/riffle, run/bend ratio
Tertiary — Riparian and Bank Structure
7. Bank stability*^
8. Bank vegetation^'.
9. Streamside cover
Subtotal for tertiary parameters
Score =
Proportion (%) of ecoregional reference
Classification =

C(D

14
18
16

7
10
11

6
9
10
25
101
81
S
Stations
3

8
6 (18)
9

2
4
10

7 (10)
9 (10)
8 (10)
24 (30)(b)
63 (81)
50 (65)
P

4

18
10
18

11
13
11

9
10
8
27
108
86
S

R(6)

18
18
19

13
13
14

10
10
10
30
125
100
E
        Criteria:

          > 90% excellent (comparable to reference)
            75-89  supporting
            60-74  partially supporting
          < 59 nonsupporting
        (a)  Reversible habitat alteration (RHA) parameters.
        (b)  Parentheses indicate adjustment for RHAs pertinent to ecoregional reference.

-------
                      TABLE 8.4-2  SUMMARY OF METRIC VALUES, PERCENT COMPARISON, AND BIOASSESSMENT SCORES FOR
                                   ARARAT AND MITCHELL RIVERS BENTHIC CASE STUDY DATA





Metric Value
Station
Metrics
Taxa richness

HBI
Scrapers/Filt. Collect.
EPT/Chiron. Abundance
% Contrib. Dom. Taxon
EPT Index
Community Loss Index
C( 1 )
26

4.46
0.833
2.45
11.2
12
0.64
3
11

9.34
0.000
0.00
53.5
0
2.31
4
34

6.24
0.108
0.55
16.5
12
0.62
R(6)
34

3.93
1.500
9.28
14.2
14
0 .00
100-Organism Subsample
% Comparison to Reference
Station
C( 1 ) 3 4 R(6)
76 32 100 100
t a \ ( a ) 1 A \ fat
88|a) 42(a) 63
56 0 7 100
26 0 6 100
__<"> __<»> __ __<»>
86 0 86 100
(c) (c) (c ) (c)




Bioassessment Score
Station
C( 1 )
4

6
6
2
6
4
4
3
0

0
0
0
0
0
2
4
6

2
0
0
6
4
4
R(6)
6

6
6
6
6
6
6
Total score

Biological condition
    32     2    22     T2

Slight  Sev.  Mod.    Non
(a)  HBI comparison is ratio of reference to station evaluated.
(b)  Actual percent contribution is evaluated, not percent comparability.
(c)  Range of values is evaluated, not percent comparability.

-------
   It is apparent from a general comparison of habitat
quality and biological condition at this North Carolina
site that there is a close relationship between habitat
and biological condition (Figure 8.4-1). As habitat
quality declines, so does the value of the  benthic
index (based on the RBP approach). If these data
from the pilot study are plotted against the theoretical
curve depicting  the relationship between habitat and
biological condition,  the deviation from the predicted
relationship for each station can be discerned (Fig-
ure 8.4-2). The development of a substantial reference
database would allow for the development of an
empirical line  with statistical confidence intervals
around the line. From this information, predictions  of
water quality effects beyond the  habitat constraints are
possible. In this manner, cause of the degradation of
biological condition at Stations 3 and 4 could be
refined.
                                                                                         125
                                                                                      - 100
              0)
              m
              OL
              m
              tr
                                       a
                                       s
                                       'E
                                       a
     Figure 8.4-1.  The relationship between habitat quality and benthic community condition at the North Carolina
                  pilot study site.
              100 —
                    0       10      20      30       40      50      60      70       80      90      100
                                          Habitat Quality (% of Reference)

            Figure 8.4-2.  Pilot study results applied to the theoretical habitat vs. biological condition curve.
                                                      8-19

-------
                                         REFERENCES
American Public Health Association, American Water-
works Association, and Water Pollution Control Feder-
ation. 1971. Standard Methods for the Examination of
Water and Wastewater. American Public Health
Association, Washington, D.C.

Angermeier, PL.  1983. The Importance of Cover and
other Habitat Features to the Distribution and Abun-
dance of Illinois Stream Fishes. Ph.D. Dissertation,
University of Illinois, Urbana.

Angermeier, PL.  and J.R. Karr. 1986. Applying an
index of biotic integrity based on  stream fish commu-
nities: Considerations in sampling and interpretation.
N. Am. J. Fish. Manage. 6:418-429.

Bailey, R.G. 1976. Ecoregions of the United States
(Map scale 1:7,500,000). USDA, Forest Service,
Ogden,  Utah.

Ball, J.  1982. Stream Classification Guidelines for
Wisconsin. Wisconsin Department of Natural
Resources Technical  Bulletin. Wisconsin Department
of Natural Resources, Madison, Wisconsin.

Benke, A.C., T.C. Van Arsdall,  Jr., and D.M.
Gillespie.  1984. Invertebrate productivity in a subtrop-
ical blackwater  river: The importance of habitat and
life history. Ecol.  Monogr.  54(l):25-63.

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, Environ-
mental Protection Agency, Marion, Illinois.

Bode, R.W.  1988.  Quality Assurance  Work Plan
for Biological Stream Monitoring  in New York State.
New York State Department of  Environmental
Conservation.

Bond, C.E.  1986.  Department of Fisheries and Wild-
life,  Oregon-State University, Corvallis.  Personal
communication.

Boesch, D.F. 1977. Application  of Numerical Classifi-
cation in Ecological  Investigation  of Water Pollution.
Report No. EPA-600/3-77-033. U.S. EPA, Corvallis,
Oregon.

Bray, J.R. and J.T. Curtis. 1957. An ordination of the
upland forest communities of southern Wisconsin.
Ecol. Monogr. 27:325.
Cairns, J., Jr.  1982. Artificial Substrates. Ann Arbor
Science Publishers, Inc., Ann Arbor, Michigan.

Cairns, J., Jr.  and K.L. Dickson. 1971. A simple
method for the biological assessment of the effects of
waste discharges on aquatic bottom-dwelling organ-
isms. J. Water Pollut.  Control Fed. 43:755-772.

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.

Clements, W.H.,  D.S. Cherry, and J. Cairns,  Jr.  1988.
Structural alterations in aquatic insect communities
exposed to copper in  laboratory streams. Environ.
Tox. and Chem. 7(9):715-722.

Cochran, W.G. 1952.  The x2 test of goodness of fit.
Ann. Math.  Statistics  23:315-345.

Courtemanch, D.L. and S.P Davies. 1987. A  coeffi-
cient of community loss to assess detrimental change
in aquatic communities. Water Res. 21(2):217-222.

Cummins, K.W. 1973.  Trophic  relations of aquatic
insects. Ann. Rev. of  Entomol.  18:183-206.

Cummins, K.W. 1987. Appalachian Environmental
Laboratory, University of Maryland, Frostburg. Per-
sonal communication.

Cummins, K.W. and M.J. Klug.  1979. Feeding ecology
of stream invertebrates.  Ann.  Rev.  Ecol.  Syst.
10:147-172.

Cummins, K.W. and M.A. Wilzbach. 1985. Field
Procedures for Analysis of Functional Feeding Groups
of Stream Macroinvertebrates.  Contribution 1611.
Appalachian Environmental Laboratory,  University of
Maryland, Frostburg.

Cummins, K.W.,  M.A. Wilzbach, D.M. Gates, J.B.
Perry, and W.B. Taliaferro. 1989. Shredders and ripar-
ian vegetation.  Bioscience. 39(1):24-30.

Dawson,  C.L. and R.A. Hellenthal.  1986. A  Com-
puterized System  for the Evaluation of Aquatic
Habitats  Based on Environmental Requirements and
Pollution Tolerance Association of Resident Organ-
isms. Report  No.  EPA-600/53-86/019. U.S.  EPA, Cin-
cinnati, Ohio.

Dimick,  R.E.,  and F.  Merryfield.  1945. The  Fishes of
the Willamette  River System in Relation to  Pollution.
                                                    R-l

-------
Engineering Experiment Station Bulletin Series
20:7-55.  (Oregon State College, Corvallis, Oregon).

Energy, Mines, and Resources Canada. 1986.  Canada
Wetland  Regions  (Map scale 1:7,500,000). MCR 4108.
Canada Map Office, Energy, Mines, and Resources
Canada,  Ottawa,  Ontario.
Fausch, D.D., J.R. Karr, and  P.R. Yant. 1984.
Regional application of an  index of biotic integrity
based on stream  fish communities. Trans. Am. Fish.
Soc. 113:39-55.
Ferrington, L.C.  1987.  Collection and Identification of
Floating  Exuviae of Chironomidae for Use in Studies
of Surface Water  Quality. SOP No. FW 130A. U.S.
EPA, Region VII, Kansas City, Kansas.
Funk, J.L.  1957.  Movement of stream  fishes  in Mis-
souri. Trans. Am. Fish.  Soc.  85:39-57.

Furse, M.T., D. Moss, J.F. Wright, and P.D.
Armitage. 1984. The influence of  seasonal and taxo-
nomic factors on  the ordination and classification of
running-water sites in Great Britain and on the predic-
tion of their macro-invertebrate communities. Fresh-
water Biol. 14:257-280.
Gabler, R.E., R.J. Sager, S. Brazier, and D.L. Wise.
1976. Essentials of Physical Geography. CBS  College
Publishing, The Dryden  Press, New York, N.Y.
Gammon, J.R. 1980. The use  of community param-
eters derived from electrofishing catches of river fish
as indicators of environmental quality,  in Seminar on
Water Quality Management Tradeoffs.  Report  No.
EPA-905/9-80-009. U.S.  EPA, Washington, D.C.
Gammon, J.R. 1989. Department of Biological
Science,  DePauw University, Greencastle, Indiana.
Personal communication.
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 Ecolog-
ical  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 for Testing and Materials,  Philadelphia,
Pennsylvania.
Gauch,  H., Jr. 1982.  Multivariate  Analysis in Com-
munity Ecology.  Cambridge University Press, New
York.
Gerking, S.D. 1959. The restricted movement  of fish
populations. Biol. Review.  34:221-242.
Hargrove, B.T.  1972. Aerobic decomposition of sedi-
ment and detritus as a function of particle surface
area and organic content. Limnol. Oceanogr.
17:583-596.
Hendricks, M.L., C.H. Hocutt, and J.R. Stauffer, Jr.
1980. Monitoring of fish in lotic habitats, in Biologi-
cal Monitoring of Fish (C.H. Hocutt and J.R.
Stauffer, Jr., eds.).  D.  C. Heath Co., Lexington,
Massachusetts.

Hill, M.O. 1979. DECORANA: A FORTRAN  Pro-
gram for Detrended Correspondence Analysis and
Reciprocal Averaging. Cornell University, Ithaca, New
York.

Hill, J.  and G.D.  Grossman.  1987. Home range esti-
mates for three North American stream fishes. Copeia
1987:376-380.

Hilsenhoff, W.L.  1982. Using a Biotic  Index to Evalu-
ate Water Quality in Streams.  Technical Bulletin
No.  132. Department of Natural  Resources,  Madison,
Wisconsin.

Hilsenhoff, W.L.  1987a. Department of Entomology,
University of Wisconsin-Madison. Personal
communication.
Hilsenhoff, W.L.  1987b. An improved biotic index
of organic stream pollution.  Great Lakes Entomol.
20:31-39.
Hilsenhoff, W.L.  1988. Rapid  field assessment of
organic  pollution with  a family level biotic  index.
J.  N. Am. Benthol. Soc. 7(l):65-68.
Hocutt,  C.H. 1981.  Fish as indicators of biological
integrity. Fisheries 6(6):28-31.
Holden,  P.B.  and C.B.  Stalnaker.  1975.  Distribution
and abundance  of mainstream  fishes of the  middle and
upper Colorado river basins,  1967-1973. Trans. Am.
Fish Soc. 104:217-231.
Hughes, R.M.  1985. Use of watershed  characteristics
to select control streams for estimating effects  of
metal mining wastes on extensively disturbed streams.
Environ. Manage. 9:253-262.

Hughes, R.M., J.H. Gakstatter, M.A. Shirazi.  and
J.M. Omernik.  1982. An approach for  determining
biological integrity  in flowing  waters, in Inplace
Resource  Inventories: Principles and Practices,
Proceedings of a National  Workshop (T.B. Brann,
ed.). Society  of American Foresters.  Bethesda,
Maryland.
                                                    R-2

-------
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. 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., 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.
Jaccard, P. 1912. The distribution of flora  in an alpine
zone. New Phytol. 11:37.
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,
Colorado.
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. Spe-
cial Publication 5. Illinois Natural History Survey.

Kuehne, R.A. and R.W. Barbour. 1983.  The American
Darters. University Press of Kentucky,  Lexington,
Kentucky.
Landers, D.H., J.M. Eilers, D.F. Brakke, W.S. Overton,
P.E. Kellar, M.E. Silverstein, R.D.  Schonbrod,
R.E. Crowe, R.A. Linthurst, J.M.  Omernik, S.A.
Teague, and E.P.  Meier.  1987.  Characteristics of Lakes
in the Western United  States. Volume I. Population
Descriptions and  Physico-Chemical Relationships.
Report No. EPA-600/3-86/054a. U.S. EPA, Washing-
ton, D.C.
Larsen, D.P.,  J.M. Omernik, R.M. Hughes, C.M.
Rohm, T.R. Whittier, A.J. Kinney, A.L. Gallant, and
D.R. Dudley.  1986. The correspondence between spa-
tial patterns in fish assemblages in Ohio streams and
aquatic ecoregions. Environ. 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.
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
State Museum of Natural History. Raleigh, North
Carolina.

Leidy,  R.A. and P.L.  Fiedler. 1985. Human distur-
bance and patterns of fish species diversity in the San
Francisco Bay drainage, California. Biol. Conserv.
33:247-267.

Lenat,  D.R. 1983. Chironomid taxa richness: Natural
variation and use in pollution assessment. Freshwater
Invertebr.  Biol. 2(4): 192-198.
Lenat,  D.R. 1986. Memorandum to Steve Tedder re:
Biomonitoring of Ararat River, September 1986. North
Carolina Department of Natural Resources and Com-
munity Development, Division of Environmental
Management.  1 October. Unpublished.
Leonard, P.M. and D.J. Orth. 1986. Application and
testing of an index of biotic integrity in small, cool-
water streams. Trans.  Am. Fish. Soc.  115:401-414.

Linthurst, R.A., D.H.  Landers, J.M.  Eilers,  D.F.
Brakke, W.S. Overton, E.P. Meier, and R.E. Crowe.
1986. Characteristics of Lakes in the Eastern United
States.  Volume I. Population Descriptions and
Physico-Chemical Relationships. Report No.
EPA-600/4-86/007a. U.S. EPA, Washington,  D.C.
Matthews, W.J. 1986.  Fish faunal structure in an
Ozark  stream: Stability, persistence, and a catastrophic
flood.  Copeia. 1986:388-397.
McArthur, J.V., J.R. Barnes, B.J. Hansen, and L.G.
Leff. 1988. Seasonal dynamics of leaf litter breakdown
in a Utah alpine stream. J. N. Am. Benthol. Soc.
7(1):44-50.
Merritt, R.W.  and K.W. Cummins, eds. 1984. An
Introduction to the Aquatic Insects of North America.
Second edition. Kendall/Hunt Publishing Co.,
Dubuque, Iowa.
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, D.J.  Orth.  1988a.
Regional 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 north-
eastern United States. N. American J.  Fish. Man. (In
Review).
                                                    R-3

-------
Minckley, W.L. 1973. Fishes of Arizona. Sims Print-
ing,  Phoenix, Arizona.

Minshall, G.W., K.W. Cummins,  R.C.  Petersen,  C.E.
Gushing, D.A. Bruns, J.R. Sedell, and R.L.  Vannote.
1985. Developments in stream ecosystem theory. Can.
J. Fish. Aquat. Sci. 42:1045-1055.

Moriseta, M. 1959.  Measuring of interspecific associ-
ation and similarity between communities.  Memoirs
Faculty Sci., Kyoshu Univ.  Ser. E. Biol. 3-65.

Mount, D.I., N.A. Thomas, T.J.  Norberg,  M.T. Bar-
bour, T.H. Roush,  and W.F. Brandes.  1984. Effluent
and  Ambient Toxicity Testing and Instream Commu-
nity  Response on the Ottawa River, Lima,  Ohio.
Report No.  EPA-600/3-84-080. U.S.  EPA,  Duluth,
Minnesota.

Mount, D.I., T.J. Norberg-King,  and A.E. Steen.
1986. Validity of Effluent and Ambient Toxicity  Tests
for Predicting Biological Impact, Naugatuck  River,
Waterbury, Connecticut. Report No. EPA-600/8-86/005.
U.S.  EPA,  Duluth,  Minn.

Moyle, P.B. 1976. Inland Fishes of California. Univer-
sity  of California Press, Berkeley, California.

Moyle, P.B., and R.D. Nichols. 1973. Ecology of
some native and introduced fishes of the Sierra
Nevada foothills in central California, Copeia.
1973:478-490.

Newman, R.M., J.A. Perry, E. Tam, and R.L. Craw-
ford. 1987. Effects of chronic chlorine  exposure  on lit-
ter processing in outdoor  experimental streams.
Freshwater Biol. 18:415-428.

Nielsen,  L.A. and  D.L. Johnson, eds.  1983.  Fisheries
Techniques. American Fisheries Society,  Bethesda,
Maryland.

North Carolina Department of Natural Resources and
Community Development. 1983. Qualitative Sampling
of Benthic  Macroinvertebrates: A Reliable, Cost-
Effective Biomonitoring Technique. Biological Series
No.  108. North Carolina Department of Natural
Resources and Community Development, Division of
Environmental Management, Raleigh, North  Carolina.

Nuzzo, R.  1986. Macroinvertebrate Rapid Assessment
Methodology. Mass. Viv.  WPC.

Ohio Environmental Protection Agency. 1987a. Biolog-
ical  Criteria for the Protection of Aquatic Life: Vol-
ume I. The Role of Biological Data  in Water Quality
Assessment (Final Draft). Ohio Environmental Protec-
tion  Agency, Columbus, Ohio.

Ohio Environmental Protection Agency. 1987b. Biolog-
ical  Criteria for the Protection of Aquatic Life: Vol-
ume II. User's Manual for Biological Assessment of
Ohio Surface Waters.  Ohio Environmental Protection
Agency, Columbus, Ohio.
Ohio Environmental Protection Agency.  1987c. Biolog-
ical Criteria for the Protection of Aquatic Life:  Vol-
ume III.  Standardized Biological Field Sampling and
Laboratory Methods for Assessing Fish and Macroin-
vertebrate Communities. Ohio Environmental  Protec-
tion  Agency, Columbus, Ohio.

Omernik, J.M.  1987. Ecoregions of the Conterminous
United States. Ann. Assoc. Am. Geograph. 77(1): 118-125.
Osborne, L.L. and E.E.  Hendricks. 1983. Streamflow
and Velocity as Determinants of Aquatic Insect  Distri-
bution  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 WE. Barber.  1982. Assessment of
fish habitat in streams: Goals, constraints, and a new
technique. Fisheries 7(3): 8-11.
Palmer, C.M. 1977. Algae and Water Pollution.  Report
No. EPA-600/9-77-036. U.S.  EPA, Cincinnati, Ohio.
Patrick, R. 1973. Use  of algae, especially diatoms in
the assessment of water quality, in Biological  Methods
for the Assessment of Water Quality (J. Cairns, Jr.
and K.L. Dickson, eds.), Special  Technical Publica-
tion 528. American Society for  Testing and Materials,
Philadelphia, Pennsylvania.
Pinkham, C.F.A. and  J.B. Pearson. 1976.  Applications
of a  new coefficient of similarity to pollution  surveys.
J. Water Pollut. Control Fed. 48:717-723.
Platts,  W.S. 1974.  Geomorphic and aquatic conditions
influencing salmonids and stream  classification—with
application to ecosystem management. U.S. Depart-
ment of Agriculture SEAM Program, Billings,
Montana.
Platts,  W.S., W.F. Megahan, and 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, Utah.

Pollard, J.E.  1981.  Investigator differences associated
with a  kicking method for sampling macroinver-
tebrates. J. Freshwater Ecol. 1:215-224.
Rankin, E. 1987. Ohio Environmental Protection
Agency. Personal communication.
Resh, V.H. 1988.  Variability, accuracy, and taxonomic
costs of rapid assessment approaches in benthic
biomonitoring. Presented at the 36th annual North
                                                   R-4

-------
American Benthological Society meeting at Tus-
caloosa, Alabama, 17-20 May 1988.
Reynolds, J.B. 1983. Electrofishing, in Fisheries Tech-
niques (L.A. Nielsen and D.L. Johnson, eds.),
American Fisheries Society, Bethesda, Maryland.
Rodgers, J.H., Jr., K.L. Dickson, and J. Cairns, Jr.
1979. A review and analysis of some methods used to
measure functional aspects of periphyton, in  Methods
and Measurements of Periphyton Communities: A
Review (R.L. Weitzel, ed.), Special Technical Publi-
cation  690. American Society for Testing and
Materials.
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, ST., W.J. Matthews, and A.E. Echelle. 1985.
Persistence of stream fish assemblages: Effects of
environmental change. Am. Nat. 126:24-40.
Scott,  W.B.  and E.J. Grossman. 1973.  Freshwater
Fishes of Canada.  Fisheries Resource Board  of
Canada, Bulletin 184.

Seber,  G.A. 1982.  The Estimation of Animal Abun-
dance. McMillan Publishing, New York, N.Y.

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.
Shackleford, B.  1988. Rapid Bioassessments of Lotic
Macroinvertebrate Communities: Biocriteria Develop-
ment. Arkansas Department of Pollution Control and
Ecology, Little Rock, Arkansas.
Simpson,  J.C. and R.L. Wallace. 1982. Fishes of
Idaho. University  Press of Idaho. Moscow, Idaho.
Snedecor, G.W.  and W.G. Cochran.  1967. Statistical
Methods.  Iowa State University  Press, Ames, Iowa.
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.

Swift,  M.C., K.W. Cummins, and R.A. Smucker.
1988a. Effects of  Dimilin on stream leaf-litter process-
ing rates..Verh. Internal. Verein. Limnol. 23:1255-1260.
Swift, M.C., R.A. Smucker, and K.W. Cummins.
1988b. Effects of  Dimilin on freshwater litter decom-
position. Environ.  Toxic, and Chem. 7:161-166.
U.S. Department of Agriculture, Soil Conservation
Service. 1981. Land Resource Regions and Major
Land Resource Areas of the United States. Agricul-
tural Handbook 296. U.S.  Government Printing
Office, Washington, D.C.
U.S. Environmental Protection Agency (EPA).  1980a.
Guidelines and Specifications for Preparing Quality
Assurance Program Plans. Report No. QAMS-004180.
U.S. EPA, Washington,  D.C.
U.S. Environmental Protection Agency (EPA).  1980b.
Interim Guidelines and Specifications for Preparing
Quality Assurance Project Plans. Report No. QAMS-
005180. U.S.  EPA, Washington, D.C.
U.S. Environmental Protection Agency (EPA).  1983.
Technical  Support Manual: Waterbody Surveys and
Assessments  for Conducting Use Attainability
Analyses.
U.S. Environmental Protection Agency (EPA).  1984a.
Policy and Program Requirement to Implement the
Quality Assurance Program. EPA Order 5360.1. U.S.
EPA, Washington, D.C.
U.S. Environmental Protection Agency (EPA).  1984b.
The Development of Data Quality Objectives. Pre-
pared by the  EPA Quality Assurance Management
Staff and the DQO Workgroup. U.S. EPA, Washing-
ton, D.C.
U.S. Environmental Protection Agency (EPA).  1984c.
Guidance  for Preparation  of Combined Work/Quality
Assurance Project Plans for Environmental Monitor-
ing. Report No. OWRS  QA-1. U.S. EPA, Washington,
D.C.
U.S. Environmental Protection Agency (EPA).  1985.
Technical  Support Document for Water Quality-based
Toxics Control. Report  No. 440/4-85-03. Office of
Water, U.S. EPA, Washington, D.C.
U.S. Environmental Protection Agency (EPA).  1987.
Surface Water Monitoring: A Framework for Change.
Office of  Water, Office  of Policy Planning and Evalua-
tion, U.S. EPA, Washington, D.C.
U.S. Environmental Protection Agency (EPA).  1988.
Proceedings of the First National Workshop on Bio-
logical Criteria, Lincolnwood, Illinois, December 2-4,
1987. Report  No. 905/9-89/003. U.S. EPA, Chicago,
Illinois.
Van Deventer, J.A.  and  W.S.  Platts. 1989.  Microcom-
puter Software System for Generating Population
Statistics from  Electrofishing Data-User's Guide for
MicroFish 3.0.  Technical Report No. INT-254.  U.S.
Department of Agriculture, U.S. Forest Service,
Ogden, Utah.
                                                    R-5

-------
Vermont Department of Environmental Conservation.
1987. Compliance Monitoring of the Aquatic Biota.
Report No. 388-1330-RJ10.
Wade, D.C. and S.B. Stalcup. 1987. Assessment of the
Sport Fishery Potential for the Bear Creek Floatway:
Biological Integrity of Representative Sites, 1986.
Report No. TVA/ONRED/AWR-87/30. Tennessee Val-
ley Authority, Muscle Shoals, Alabama.
Warren, C.E.  1979.  Toward Classification and Ration-
ale for Watershed Management and Stream Protection.
Report No. EPA-600/3-79-059. U.S. Environmental
Protection Agency, Corvallis, Oregon.
Weitzel, R.L. 1979. Periphyton measurements and
applications,  in Methods and Measurements of
Periphyton Communities:  A Review (R.L. Weitzel,
ed.), Special Technical Publication 690. American
Society for Testing and Materials.
Whittaker, R.H. 1952. A  study of summer foliage
insect communities  in the Great Smokey Mountains.
Ecol. Monogr. 22:6.
Whittier, T.R., D.P. Larsen, R.M. Hughes, CM.
Rohm, A.L. Gallant, and J.M. Omernik. 1987. The
Ohio Stream Regionalization Project: A Compendium
of Results.  Report No.  EPA-600/3-87/025. U.S. EPA,
Corvallis, Oregon.
Whittier, T.R.,  R.M. Hughes, and D.P. Larsen. 1988.
Correspondence between ecoregions and spatial pat-
terns in stream  ecosystems in Oregon. Can. J. Fish.
Aquat. Sci. 45:1264-1278.
Winget, R.N. and F.A. Mangum. 1979. Biotic Condi-
tion Index:  Integrated Biological, Physical, and Chem-
ical Stream Parameters  for Management. Intermountain
Region, U.S. Department of Agriculture, Forest Ser-
vice, Ogden, Utah.
Winner, R.W., M.W. Boesel, and M.P. Farrell. 1980.
Insect community structure as an index of heavy-metal
pollution in lotic ecosystems. Can. J. Fish. Aquat.
Sci. 37:647-655.
Wydoski, R.S. and R.R. Whitney. 1979. Inland Fishes
of Washington.  University  of Washington Press.
Seattle, Washington.
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     APPENDIX A

 GUIDANCE FOR USE OF FIELD
AND LABORATORY DATA SHEETS

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                                      APPENDIX A
                       GUIDANCE FOR USE  OF FIELD
                     AND LABORATORY  DATA SHEETS
   This appendix provides guidance for use of the
rapid bioassessment field and laboratory data sheets.
The guidance sheets give brief descriptions of the
information required for each data sheet.
A.2  GUIDANCE FOR BIOSURVEY
      FIELD DATA  SHEET FOR
   BENTHIC RBPs I,  II, AND III
    (Figures 6.1-1, 6.2-1, and 6.3-1)
   A.I GUIDANCE FOR HEADER
    INFORMATION (Figure 2.6-1)
Water body Name:
  Name of river,  stream, or drain.
Location:
  Township, range, section, county where problem
  area is located. For rivers, streams, or drains, road
  crossings or outfall locations should be referenced
  where applicable.
Reach/Milepoint:
  Indicate station reach/milepoint.
Latitude/Longitude:
  Indicate station latitude/longitude.
County/State:
  Name of county and state where station is located.
Aquatic Ecoregion:
  Name of ecoregion.
Station:
  Agency name or number for station.
Investigators:
  List field personnel involved.
Date:
  Date of survey.
Agency:
  Agency name or affiliation (academic, private
  consulting)
Hydrologic Unit Code:
  Indicate the USGS cataloging unit  number in which
  the station is located.
Form Completed By:
  List personnel completing form.
Reason for Survey:
  Reason survey was conducted.
    Rapid Bioassessment Protocol I
               (Figure 6.1-1)

Estimated Abundance Level of Aquatic Biota:
Record estimated abundance level  of biota found in
the sampling area. Circle the number (corresponding
to the descriptions just below on the data sheet) that
best indicates the estimated size of each population
found in the sampling area. Each  agency should
develop its own abundance level criteria.

Macrobenthos Qualitative Sample: Using the guide-
lines of rare, common, abundant,  or dominant, record
the estimated abundance level of each major taxa
found in the sampling area. Each  agency should
develop its own abundance level criteria.

Observations: Information included here would
include abundance of fish nesting  sites;  notes concern-
ing biota present; type of game fish observed; location
or presence  of noteworthy  physical structures such as
bridges, rip-rap, culverts; habitat alteration due to
construction activities;  or any other observations perti-
nent to an impact assessment.

   Rapid Bioassessment Protocol  II
               (Figure 6.2-1)

Estimated Abundance Level of Aquatic Biota:
Record estimated abundance level  of biota found in
the sampling area. Circle the number (corresponding
to the descriptions just below on the data sheet) that
best indicates the estimated size of each population
found in the sampling area. Each  agency should
develop its own abundance level criteria.
                                               A-l

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Macrobenthos Qualitative Sample List: List families
found in 100-organism subsample and the number of
individuals within each family.

Riffle Sample Functional Feeding Groups:
Record the number of individuals collected  in the
100-organism  riffle  subsample which represent the
Scraper and Filtering Collector Functional Feeding
Groups.

CPOM Sample: Record the number of individuals
collected in the supplemental CPOM sample which
represent the Shredder Functional Feeding Group, and
total number of individuals sorted.

Observations: Information included here would
include abundance of fish  nesting sites; notes concern-
ing biota present; type of game fish observed; location
or presence of noteworthy  physical structures such as
bridges, rip-rap, culverts; habitat alteration due to
construction activities; or any other observations perti-
nent to an  impact assessment.
   Rapid Bioassessment Protocol III
                (Figure 6.3-1)

Estimated Abundance Level of Aquatic Biota:
Record estimated abundance level of biota found in
the sampling area. Circle the number (corresponding
to the descriptions just below on the data sheet) that
best indicates  the estimated size of each population
found in the sampling area. Each agency should
develop its own abundance level criteria.

Macrobenthos Qualitative Sample: Using the guide-
lines of rare, common, abundant, or dominant, record
the estimated abundance level of each major taxa
found in the sampling area. Each agency should
develop their own abundance level criteria.

CPOM Sample: Record the number of individuals
collected in the supplemental CPOM sample which
represents the Shredder Functional Feeding Group,
and total number of individuals sorted.

Observations: Information recorded here would
include abundance of fish nesting sites;  notes concern-
ing biota present; type of game fish observed; location
or presence of noteworthy  physical structures such as
bridges, rip-rap, culverts; habitat alteration due to
construction activities;  or any other observations perti-
nent to an impact assessment.
A.3  GUIDANCE FOR IMPAIRMENT
     ASSESSMENT SHEET FOR
         RBPs  I,  II, III, AND V
         (Figures 6.1-2  and 7.2-1)
 Rapid Bioassessment Protocols I,  II,
                 III,  and V

 1.  Detection of Impairment:  Circle the one that
   applies.
2.  Biological Impairment Indicator: Circle those
   that apply, as indicated by the benthos, fish,  and
   other aquatic biota.
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).
4.  Cause: Circle  those  that apply. Indicate which  is
   the major cause of the stream problem.
5.  Estimated Areal Extent of Problem: Record esti-
   mated downstream extent of impact (in m) and
   multiply  by approximate stream width (in m) to
   estimate  areal width.
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.
     A.4  GUIDANCE FOR  DATA
       SUMMARY SHEET FOR
     BENTHIC RBPs  II AND III
         (Figure 6.2-2 and  6.3-3)
   Rapid  Bioassessment Protocol II
               (Figure 6.2-2)

Station Number: Indicate station number for each
data set recorded.

Station Location: Record brief description of sam-
pling site relative to established landmarks (i.e.,
roads, bridges).
                                                 A-2

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Taxa Richness: Record total number of families (or
higher taxa) collected in the 100-organism riffle
subsample.

FBI (modified): Record the Family Biotic Index value
(Hilsenhoff 1988) calculated for the 100-organism rif-
fle subsample using the formula presented in RBP II.
Tolerance  classification values can be entered into  the
computer database to simplify calculation.

Functional Feeding Group: Functional Feeding
Group classifications may be entered into the com-
puter  database to  simplify calculations.

   Riffle Community; Scrapers/Filtering Collectors:
   Enter the value obtained by  dividing the number of
   individuals in the riffle subsample representing  the
   Scraper Functional Group, by the number repre-
   senting the Filtering Collector Functional Group.

   CPOM Community; Shredders/Total: Enter the
   value obtained by dividing the number of
   individuals in the CPOM sample (or subsample)
   representing the Shredder Functional Group, by the
   total number of organisms in the sample (or
   subsample).

EPT/Chironomidae: Enter the value obtained by
dividing the number of individuals in the
100-organism riffle subsample in the family
Chironomidae, by the total  number of individuals in
the orders Ephemeroptera,  Plecoptera, and
Trichoptera.

Percent Contribution (Dominant Family): Record
the value obtained by dividing the number of individ-
uals in the family that is most  abundant in the
100-organism riffle subsample, by the total number of
individuals in the sample.

EPT Index: Record the total number of taxa in the
100-organism riffle subsample representing the  orders
Ephemeroptera, Plecoptera, and Trichoptera.

Community Similarity Index: Enter the value calcu-
lated for the appropriate community similarity index,
using  data from the 100-organism riffle subsample.
   Values obtained for each metric should be assigned
a score based on  percent comparability to the control
or reference station data. Scores are summed for both
the impaired and  reference  station. The percent com-
parison between  the total  scores provides the final
evaluation of biological condition.
   Rapid Reassessment Protocol III
                (Figure 6.3-3)

Station Number: Indicate station number for each
data set recorded.

Station Location: Record brief description of sam-
pling site relative to established landmarks (i.e.,
roads,  bridges).

Species Richness: Record total number of species (or
higher  taxa) collected in the 100-organism riffle
subsample.

HBI (modified): Record the species level Hilsenhoff
Biotic  Index value (Hilsenhoff 1987b) calculated for
the 100-organism riffle subsample using the formula
presented in Rapid Bioassessment Protocol III. Toler-
ance classification values can be entered into the com-
puter database to simplify calculation.

Functional Feeding Group: Functional Feeding
Group  classifications may be entered into the com-
puter database to simplify calculations.

   Riffle Community; Scrapers/Filtering Collectors:
   Enter the value obtained by dividing the number of
   individuals  in the  riffle subsample representing the
   Scraper  Functional Group, by the number repre-
   senting the  Filtering Collector Functional Group.

   CPOM  Community; Shredders/Total: Enter the
   value obtained by  dividing the number of indi-
   viduals in the CPOM sample (or subsample)
   representing the Shredder Functional Group, by the
   total number of organisms in the sample (or
   subsample).

EPT/Chironomidae: Enter the value obtained  by
dividing the total number of individuals in the
100-organism riffle subsample in the orders
Ephemeroptera,  Plecoptera, and Trichoptera, by the
number of individuals in the family Chironomidae.

Percent Contribution (Dominant Taxon): Record the
value obtained by dividing the number of individuals
in the  taxon that is most abundant in the 100-organism
riffle subsample, by the total  number of individuals in
the sample.

EPT Index: Record the total number of taxa in the
100-organism riffle subsample representing the orders
Ephemeroptera,  Plecoptera, and Trichoptera.
                                                    A-3

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Community Similarity Index: Enter the value calcu-
lated for the appropriate community similarity index,
using data from the 100-organism riffle subsample.
   Values obtained for each metric should be assigned
a score based on percent comparability to the control
or reference station data. Scores are summed for both
the impacted and reference station.  The percent com-
parison between the total scores provides the final
evaluation of biological condition.
   Note: To maximize time efficiency, the metric
         values entered on the Data Summary Sheets
         are intended to be generated by use of a
         computerized database.
  A.5  GUIDANCE FOR LABORA-
     TORY BENCH SHEET FOR
            BENTHIC RBP III
                (Figure 6.3-2)

Station Number: Indicate agency assigned number for
each sample dataset recorded.

Station Location: Record  brief description of sam-
pling site relative to established landmarks (i.e.,
roads,  bridges).

Species: List species identified from the 100-organism
riffle subsample.

Number  of Organisms: Indicate number of indi-
viduals of each species collected in the  100-organism
riffle subsample at each station.

Total Organisms: Record  total number  of individuals
collected  in the 100-organism riffle subsample at each
station.

Number  of Taxa: Record  the total number of taxa
collected  in the 100-organism riffle subsample at each
station.
     A.6 GUIDANCE FOR FIELD
 COLLECTION DATA  SHEET FOR
                FISH  RBP  V
                (Figure 7.2-3)

Drainage: Give name of stream or river and its basin,
site descriptor,  and unique site code.
Date: Enter day, month, and year of collection.

Sampling Duration: Record length of time in minutes
actually collecting fish.  If replicates are taken, record
them separately.

Sampling Distance: Measure,  with a tape or cali-
brated range finder, the length  in meters of reach
sampled.

Sampling Area: Multiply the length or reach sampled
by the average width sampled.  Express in meters
squared.

Crew: Indicate crew chief and crew members.

Habitat Complexity/Quality: Circle the descriptor
that best describes subjective evaluation of the phys-
icochemical habitat.

Weather: Record air temperature, estimated wind
velocity, percent cloud cover, and precipitation.

Flow: Circle most appropriate  descriptor.

Gear Used:  Specify type, model, and number of elec-
trofisher, mesh size and length of seine, or concentra-
tion of fish  toxicant.

Gear/Crew Performance: Indicate effectiveness  of
crew in sampling the site. Note problems with equip-
ment, staff, or site obstacles, such as extensive cover,
high velocity current, excessive turbidity, floating
debris, deep muck or pools, or weather conditions.

Comments: Record any additional qualitative site
data: sketch map or photographs, presence of springs,
evidence of fishing activity, any potential or current
impacts, weather conditions (such as evidence  of
recent high  flows or unusually hot or cold weather
immediately preceding the survey), biota observed
(insect hatches,  potential vertebrate predators, fish
nesting and grazing sites, fish reproductive condition,
fish seen but not captured).

Fish (preserved): Indicate if specimens were
preserved for permanent collection or further
examination.

Number of Individuals; Number of Anomalies:
Give total numbers of fish and anomalies for the
sample.

Genus/Species: Enter scientific name or unique  stan-
dard abbreviation for each species captured.
                                                   A-4

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Adults (Number, Weight): Enter the number of adults
of each species and their total weight in grams. Weigh
individually or by batch, depending on  the species'
size and abundance. Species weight can also be deter-
mined by weighing a subsample of individuals (20-30
fish spanning the size range collected) and extrapolat-
ing for the total  number of that species.

Juveniles  (Number, Weight): Record the number of
juveniles of each species and their total weight as
above. Juveniles and adults are distinguished subjec-
tively by coloration and size; the objective is to deter-
mine  whether both age classes are present.

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.
      A.7  GUIDANCE  FOR DATA
    SUMMARY SHEET FOR FISH
                     RBP V
                 (Figure 7.2-5)
Station Number: Indicate station number.

Station Location: Record brief description of sam-
pling site relative to established landmarks (i.e.,
roads, bridges).
Metrics: List metrics used to conduct IBI calcula-   -
tions. Use Karr's original metrics or published (or
well supported) substitutes. Precede metric selection
with analysis of reference site data or a high quality
historical database from a representative,  large river
basin.

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.

Metric Value: Record  metric values (number or per-
cent) for the station. Metric values are obtained by
comparing the collection data (Figure 7.2-3)  with the
tolerance and trophic guilds previously listed (e.g.,
Appendix D).  For taxonomic metrics numbers of spe-
cies are added. Total number of individuals is
recorded from the field collection data sheet.  Propor-
tional metrics  are determined by adding the number of
individuals in  each category and then dividing by the
total number of individuals.

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).

Scorer: Enter scorer's name.

IBI Score: The'metric scores (and 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.

Comments: Metrics producing contrary results or
suggestions for improvement are entered here.
                                                    A-5

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Waterbody Name.
Reach/Milepoint _
County	
 State.
                            Location.
Latitude/Longitude
Aquatic Ecoregion _
Station Number.
Date	
Hydrologic Unit Code.
Reason for Survey	
Time
Investigators.
Agency	
                            Form Completed by.
          Figure 2.6-1.  Header information used for documentation and identification for sampling stations.

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                             Rapid Bioassessment Protocol I

                                   Biosurvey Field Data Sheet
RELATIVE ABUNDANCE OF AQUATIC BIOTA
Perjphyton         0123
Filamentous Algae   0123
Macrophytes        0123


0 = Absent/Not Observed          1 = Rare
Slimes             0     1
Macroinvertebrates   0     1
Fish               0     1
                                              2 = Common
                                                                 3 = Abundant
234
234
234


  4 = Dominant
MACROBENTHOS QUALITATIVE SAMPLE LIST(lndicate Relative Abundance R = Rare, C = Common, A = Abundant, 0 = Dominant)
Porifera
Hydrozoa
Platyhelminthes
Turbellaria
Hirudinea
Oligochaeta
Isopoda
Amphipoda
Decapoda
Gastropoda
Bivalvia

Anisoptera
Zygoptera
Hemiptera
Coleoptera
Lepidoptera
Sialidae
Corydalidae
Tipulidae
Empididae
Simuliidae
Tabanidae
Culicidae
Chironomidae
Plecoptera
Ephemeroptera
Trichoptera
Other







Rare < 3
Observations
                      Common 3-9
                                                Abundant > 10
                                                                           Dominant > 50 (Estimate)
        Figure 6.1-1. Biosurvey Field Data Sheet for use with Rapid Bioassessment Protocol I.
                                              A-7

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                              Rapid  Bioassessment Protocol II

                                    Biosurvey Field Data Sheet
RELATIVE ABUNDANCE OF AQUATIC BIOTA
Periphyton
Filamentous
Macrophytes
0 1
Algae 0 1
0 1
0 = Absent/Not Observed
2
2
2
1
3
3
3
= Rare
4 Slimes
0 1
4 Macroinvertebrates 0 1
4 Fish
2 = Common
0 1
3 = Abundant
2
2
2
3
3
3
4
4
4
4 = Dominant
MACROBENTHOS QUALITATIVE SAMPLE LIST
                                                                    List Families Present/Indicate Abundance
Oligochaeta


Gastropoda


Bivalvia


Ephemeroptera




Anisoptera


Zygoptera

Plecoptera


Trlchoptera




Coleoptera


Diptera





Other
RIFFLE SAMPLE
FUNCTIONAL FEEDING GROUPS
(Indicate No. of Individuals Representing Group)
Scrapers
     Filtering Collectors
CPOM SAMPLE  FUNCTIONAL FEEDING GROUPS  (Indicate No. of Individuals Representing Group)
Shredders
                                                  Total Org. in Sample
Observations
       Figure 6.2-1. Biosurvey Field Data Sheet for use with Rapid Bioassessment Protocol II.
                                               A-8

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                             Rapid Bioassessment Protocol III

                                    Biosurvey Field Data Sheet
RELATIVE ABUNDANCE OF AQUATIC BIOTA
Periphyton         0
Filamentous Algae   0
Macrophytes        0


0 = Absent/Not Observed
1     2
1     2
1     2
                              1 = Rare
Slimes             0     1
Macroinvertebrates   0     1
Fish                0     1
                                               2 = Common
                                                                   3 = Abundant
2     34
2     34
234


  4 = Dominant
MACROBENTHOS QUALITATIVE SAMPLE LIST(lndicate Relative Abundance R = Rare, C = Common, A = Abundant, D = Dominant)
Porifera
Hydrozoa
Platyhelminthes
Turbellaria
Hirudinea
Oligochaeta
Isopoda
Amphipoda
Decapoda
Gastropoda
Bivalvia

Anisoptera
Zygoptera
Hemiptera
Coleoptera
Lepidoptera
Sialidae
Corydalidae
Tipulidae
Empididae
Simuliidae
Tabanidae
Culicidae
Chironomidae
Plecoptera
Ephemeroptera
Trichoptera
Other







Rare < 3
                      Common 3-9
                                                  Abundant > 10
                                                                             Dominant > 50 (Estimate)
CPOM SAMPLE  FUNCTIONAL FEEDING GROUPS  (Indicate No. of Individuals Representing Group)
Shredders
                                                  Total Org. in Sample
Observations
       Figure 6.3-1. Biosurvey Field Data Sheet for use with Rapid Bioassessment Protocol III.
                                                A-9

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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:

                  Benthic macroinvertebrates          Other aquatic communities

                  	 absence of EPT taxa            	 Periphyton
                  	 dominance of tolerant groups     	 filamentous

                  	 low benthic abundance            	 other

                  	 low taxa richness              	 Macrophytes

                  	 other                          	 Slimes

                                                      	 Fish


              3.   Brief description of problem: 	
                  Year and date of previous surveys:
                  Survey date, available in: 	
                  Cause: (indicate major cause)   organic enrichment    toxicants   flow

                      habitat limitations   other 	
                                                      2
                  Estimated areal extent of problem (m ) and length of  stream reach

                  affected (m), where applicable: 	
              6.  Suspected source(s) of problem:

                  	  point source discharge (name, type of facility,  location)
                  	  construction site  runoff
                  	  combined sewer outfall
                  	  silviculture runoff
                  	  animal  feedlot
                  	  agricultural runoff
                  	  urban runoff
                  	  ground  water
                  	  other
                        unknown
              Briefly explain:
Figure 6.1-2. Impairment Assessment Sheet for use with macroinvertebrate Rapid Bioassessment Protocols.
                                              A-IO

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                              IMPAIRMENT ASSESSMENT SHEET
          Detection  of  impairment:  Impairment detected
                                    (Complete Items  2-6)
             No impairment
                detected
              (Stop here)
          Biological  impairment  indicator:

            Fish

            	 sensitive  species  reduced/absent
            	 dominance  of  tolerant species
            	 skewed trophic  structure
            	 abundance  reduced/unusually high
            	 biomass reduced/unusually high
            	 hybrid or  exotic  abundance
                  unusually high
            	 poor size  class representation
            	 high incidence  of  anomalies

          Brief  description  of problem:  	
  Other aquatic communities

  	 Macroinvertebrates
  __ Periphyton
  	 Macrophytes
           Year  and  date  of  previous surveys:

           Survey  data available  in: 	
          Cause  (indicate  major  cause):  organic  enrichment    toxicants   flow

                                         sediment    temperature   poor habitat

                                         other
           Estimated areal  extent  of  problem  (m  )  and  length of stream reach

           affected (m)  where applicable: 	

           Suspected source(s) of  problem
             point  source

             urban  runoff

             agricultural runoff
             silvicultural runoff
             livestock
             landfill
mine
dam or diversion

channelization or snagging
natural
other
unknown
       Comments:
Figure 7.2-1. Impairment Assessment Sheet for use with fish Rapid Bioassessment Protocol V.
                                      A-ll

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                                         DATA SUMMARY SHEET
Station No.
Station Location
Taxa Richness
FBI (modified)
Functional Feeding Groups
Riffle Community
Scrapers/Filt. Collect.
CPOM Community
Shredders/Total
EPT/Chironomidae
% Contribution (dom. family)
EPT Index
Community Similarity Index
Comments:

















































































Figure 6.2-2.  Data Summary Sheet suggested for use in recording benthic data utilized in Rapid Bioassessment Protocol

-------
                                          DATA SUMMARY SHEET
Station No.
Station Location
Taxa Richness
FBI (modified)
Functional Feeding Groups
Riffle Community
Scrapers/Filt. Collect.
CPOM Community
Shredders/Total
EPT/Chironomidae
% Contribution (dom. family)
EPT Index
Community Similarity Index
Comments:

















































































Figure 6.3-3.  Data Summary Sheet suggested for use in recording benthic data utilized in Rapid Bioassessment Protocol III.

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                       LABORATORY BENCH SHEET
                                   Number of Organisms
Station Number
Station Location
Species Name




•










-



Total Organisms
Number of Taxa
































































































Figure. 6.3-2. Laboratory Bench Sheet suggested for use in recording benthic
             data utilized in Rapid Bioassessment Protocol III.
                                 A-14

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                           FISH FIELD COLLECTION DATA SHEET
                                                                   page
                                                                           of
      Drainage
                                           Date .
Sampling Duration (rain)
Sampling Distance (m) Sampling
Habitat Complexity/Quality (excellent
Veather Flow (floe
Gear Used Gear/Crev
Area (m ) rreu
good fair poor very poor)
d bankfull moderate lov)
Performance
Comments
Fish (preserved) Number of Individuals
Genus/Species Adults



















Number of Anomalies
Juveniles Anomalies^ '
No. tft. No. tft. No.






















































      (*) Discoloration, deformities,  eroded fins,  excessive mucus, excessive
          external parasites,  fungus,  poor condition,  reddening, tumors,
          and ulcers
Figure 7.2-3.  Fish Field Collection Data Sheet for use with Rapid Bioassessment Protocol V.
                                         A-15

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Station No.
Site






Scoring Criteria(b)
Hetrics(a)
1. Nuaber of Native Fish Species
2. Nuaber of Darter or Benthic Species
3. Nuaber of Sunfish or Pool Species
4. Nuaber of Sucker or Long-Lived Species
5. Number of Intolerant Species
6. X Green Sunfish or Tolerant Individuals
7. X Oanivores
8. X Insectivores or Invertivores
9. Z Top Carnivores
10. Total Nunber of Individuals
11. t Hybrids or Exotics
12. X Anomalies
Scorer
Coaaents:
m
>67
>67
>67
>67
>67
<10
<20
>45
>5
>67
0
<1


3
~TCT
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
TZ~) Metric Value Metric Score
<33
<33
<33
<33
<33
>25
>45
<20
<1
<33
>1
>5
IBI Score





(a) Karr's original aetrics or commonly used substitutes.
ties.
(b) Karr's original scoring criteria or coanonly
ecoregions.

See text

used substitutes.


and Table 7.2-1 for other possibili-

These nay require refinement in other

Figure 7.2-5.  Data Summary Sheet for Rapid Bioassessment Protocol V.
                             A-16

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              APPENDIX B

RAPID BIOASSESSMgNT SUBSAMPLING METHODS FOR
         BENTHIC PROTOCOLS II AND III
          (100-Organism Count Technique)

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                                      APPENDIX  B
    RAPID BIOASSESSMENT SUBSAMPLING METHODS FOR
                      BENTHIC  PROTOCOLS  II AND III
                         (100-Organism Count  Technique)
    B.1  RAPID BIOASSESSMENT
  SUBSAMPLING METHODS FOR
             PROTOCOL II
1. Thoroughly rinse sample in a (500-micron) screen
  or the sampling net to remove fine sediments. Any
  large organic material (whole leaves, twigs, algal or
  macrophyte mats) should be rinsed, visually
  inspected,  and discarded.

2. Place sample contents in a large, flat pan with a
  light-colored (preferably white) bottom. The bottom
  of the pan should be marked with a numbered grid
  pattern, each block in the grid measuring 5 cm x
  5 cm. (Sorting using a gridded pan is only feasible
  if the organism movement in the sample can be
  slowed by  the addition of club soda or tobacco to
  the sample. If the organisms are not anesthetized,
  100 organisms should be removed from the pan as
  randomly as possible.) A 30 X 45 cm pan is
  generally adequate, although pan size  ultimately
  depends on sample size.  Larger pans allow debris
  to be spread more thinly, but they are unwieldy.
  Samples too large to be effectively sorted in a sin-
  gle pan may be thoroughly  mixed in a container
  with some water, and half of the homogenized
  sample placed in each of two gridded  pans. Each
  half of the sample must be  composed  of the same
  kinds and  quantity of debris and an equal number
  of grids-must be sorted from each pan, in order to
  ensure a representative subsample.

3. Add just enough water to allow  complete dispersion
  of the sample within the pan; an excessive amount
  of water will allow sample material to shift within
  the grid during sorting. Distribute sample material
  evenly within the grid.

4. Use a random numbers table to select a number
  corresponding to a square within the gridded pan.
  Remove all organisms from within that square and
  proceed with the process of selecting squares and
  removing organisms until the total number sorted
  from the sample is within 10 percent of 100. Any
  organism which is lying over a line separating two
  squares is considered to be in the square containing
  its head. In those instances where it is not possible
  to determine the location of the head (worms for
  instances), the organism is  considered to be in the
  square containing the largest portion of its body.
  Any square sorted must be sorted in its entirety,
  even after the 100 count has been reached. In order
  to lessen sampling bias, the investigator should
  attempt to pick smaller, cryptk^ organisms  as well
  as the larger, more obvious organisms.
Source: Modified from Hilsenhoff 1987b.
    B.2  RAPID BIOASSESSMENT
  SUBSAMPLING METHODS FOR
             PROTOCOL  III
1.  Thoroughly rinse sample in a No. 35 mesh
   (500-micron) screen to remove preservative. Any
   large organic material (whole leaves, twigs, algal or
   macrophyte mats) not removed in the field should
   be rinsed, visually inspected, and discarded. If the
   samples have been preserved in alcohol, it will be
   necessary to. soak the sample contents in water for
   about 15 minutes to hydrate the benthic organisms,
   preventing them from floating on the water surface
   during sorting.

2.  Place sample contents in a large, flat pan with a
   light-colored (preferably white) bottom. The bottom
   of the pan should be marked with a  numbered grid
   pattern, each block in the grid measuring 5 cm x
   5 cm. A 30 X 45 cm pan is generally adequate,
   although pan size ultimately  depends on sample
   size. Larger pans allow debris to be  spread more
                                                B-l

-------
   thinly, but they are unwieldy. Samples too large to
   be effectively sorted in a single pan may be
   thoroughly mixed in a container with some water,
   and half of the homogenized sample placed in each
   of two gridded pans.  Each half of the sample must
   be composed of the same kinds and quantity of
   debris and an equal number of grids  must be
   sorted from each pan, in order to ensure a
   representative subsample.

3.  Add just enough  water to allow complete dispersion
   of the sample within the pan; an excessive amount
   of water will allow sample material to shift within
   the grid during sorting. Distribute sample  material
   evenly within the grid.

4.  Use a random numbers table to select a number
   corresponding to a square within the  gridded pan.
Remove all organisms from within that square and
proceed with the process of selecting squares and
removing organisms until the total number sorted
from the sample is within  10 percent of 100. Any
organism  which is lying over a line separating two
squares is considered to be in the square containing
its head. In those instances where it is not possible
to determine  the location of the head (worms for
instances), the organism is  considered to be in the
square containing the largest portion of its body.
Any square sorted must be sorted in its entirety,
even after the 100 count has been reached. If many
of the organisms are very small and it appears that
the potential  for missing individuals  is great, an
illuminated 5X magnifier will facilitate the sorting
procedure.
Source: Modified from Hilsenhoff 1987b.
                                                     B-2

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             APPENDIX C

         FAMILY AND SPECIES-LEVEL
MACROINVERTEBRATE TOLERANCE CLASSIFICATIONS

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                                     APPENDIX  C
                        FAMILY AND SPECIES-LEVEL
     MACROINVERTEBRATE TOLERANCE CLASSIFICATION
          C.1  FAMILY-LEVEL
  TOLERANCE CLASSIFICATION
    average of tolerance values of species and genera
    within each family based on their relative abun-
    dance in Wisconsin.
   RBP II is based on family-level identifications. The
adequate assessment of biological condition for RBP
II requires the use of a tolerance classification for
differentiating among responses of the benthic com-
munity to pollutants. Hilsenhoffs Family Biotic Index
(FBI) is used as a basis for the family-level tolerance
classification presented in this document.
   A brief description of the FBI is taken from
Hilsenhoffs paper entitled "Rapid Field Assessment
of Organic Pollution with a Family Biotic Index"
(Hilsenhoff 1988). The family-level tolerance values
assigned for western Great Lakes region stream
arthropods are presented in Table C-l.

  A special symposium on rapid biological assess-
  ment at the 1986 meeting of the North  American
  Benthological Society  stressed the need for rapid
  field-based assessment approaches. It was recog-
  nized that in order to  save time, a degree of
  accuracy would be sacrificed. Consequently, I
  adapted the biotic index (BI) of organic pollution
  (Hilsenhoff I987b) for rapid evaluation by providing
  tolerance values for families (Table C-l) to allow a
  family-level biotic index (FBI) to be calculated in
  the field. The FBI is an average of tolerance values
  of all arthropod families in a sample. It is not
  intended as a replacement for the  BI and can be
  effectively used in the field only by biologists who
  are familiar enough with arthropods to be able to
  identify families without using keys.

  Using the same method and more than 2,000
  stream samples from throughout Wisconsin that
  were used to revise tolerance values for species and
  genera (Hilsenhoff 1987b) family-level tolerance
  values were established by comparing occurrence of
  each family with the average BI of streams in
  which they occurred in the greatest numbers. Thus,
  family-level tolerance values tend to be a weighted
    C.2 GENUS/SPECIES-LEVEL
  TOLERANCE CLASSIFICATION
   The tolerance classification used in RBP III is
based on Hilsenhoff (1987b). Because Hilsenhoffs
tolerance classification is restricted to arthropods,
nonarthropod tolerance designations have been taken
from Bode (1988). Some of these tolerance values for
macroinvertebrates not listed in Hilsenhoff (1982,
1987b) are presented  in Table C-2.
       C.3  REFERENCES FOR
   DETERMINING FAMILY AND
   SPECIES-LEVEL TOLERANCE
          CLASSIFICATIONS
Beck, W.M., Jr. 1977. Environmental Requirements
  and Pollution Tolerance of Common Freshwater
  Chironomidae. Environmental Monitoring and Sup-
  port Laboratory, Report No. EPA-600/4-77-024.
  U.S. EPA, Cincinnati.
Bode, R.W. 1988. Quality Assurance Workplan for
  Biological Stream Monitoring in New York State.
  New York State Department of Environmental Con-
  servation, Albany, New York.
Dawson, C.L. and R.A. Hellenthal. 1986. A Com-
  puterized System for the Evaluation of Aquatic
  Habitats Based on Environmental Requirements and
  Pollution Tolerance Association  of Resident Organ-
  isms. Report No. EPA-600/S3-86/019.
Harris, T.L. and T.M. Lawrence. 1978. Environmental
                                              C-l

-------
   TABLE C-l  TOLERANCE VALUES FOR FAMILIES OF STREAM ARTHROPODS IN THE
  	WESTERN GREAT LAKES REGION  (FROM HILSENHOFF  1988)	
Plecoptera
Ephemeroptera
Odonata
Trichoptera
Megaloptera

Lepidoptera

Coleoptera

Diptera
Amphipoda

Isopoda
Capniidae 1, Chloroperlidae 1, Leuctridae 0,
Nemouridae 2, Perlidae 1, Perlodidae 2,
Pteronarcyidae 0, Taeniopterygidae 2

Baetidae 4, Baetiscidae 3, Caenidae 7, Ephemerellidae
1, Ephemeridae 4, Heptageniidae 4, Leptophlebiidae 2,
Metretopodidae 2, Oligoneuriidae 2, Polymitarcyidae
2, Potomanthidae 4, Siphlonuridae 7, Tricorythidae 4

Aeshnidae 3, Calopterygidae 5, Coenagrionidae 9,
Cordulegastridae 3, Corduliidae 5, Gomphidae 1,
Lestidae 9, Libellulidae 9, Macromiidae 3

Brachycentridae 1, Glossosomatidae 0, Helicopsychidae
3, Hydropsychidae 4, Hydroptilidae 4, Lepidosto-
matidae 1, Leptoceridae 4, Limnephilidae 4,
Molannidae 6, Odontoceridae 0, Philopotamidae 3,
Phryganeidae 4, Polycentropodidae 6, Psychomyiidae 2,
Rhyacophilidae 0, Sericostomatidae 3

Corydalidae 0, Sialidae 4

Pyralidae 5

Dryopidae 5, Elmidae 4, Psephenidae 4

Athericidae 2, Blephariceridae 0, Ceratopogonidae 6,
Blood-red Chironomidae (Chironomini) 8, other
(including pink) Chironomidae 6, Dolochopodidae 4,
Empididae 6, Ephydridae 6, Psychodidae 10, Simuliidae
6, Muscidae 6, Syrphidae 10, Tabanidae 6, Tipulidae 3

Gammaridae 4, Talitridae 8

Asellidae 8
                                    C-2

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 TABLE C-2   TOLERANCE VALUES FOR SOME MACROINVERTEBRATES NOT INCLUDED IN
	HILSENHOFF (1982,  1987b>ra7;  FROM BODE (1988)	


                  Acariformes                    A
                  Decapoda                       6
                  Gastropoda
                    Amnicola                     8
                    Bithynia                     8
                    Ferrissia                    6
                    Gyraulus~                    8
                    Helisoma                     6
                    Lymnaea                      6
                    Physa                        8
                    Sphaeriidae                  8

                  Oligochaeta
                    Chaetogaster                 6
                    Dero                        10
                    Nais barbata                 8
                    Nais behningi                6
                    Nais bretscheri              6
                    Nais communis                8
                    Nais elinguis               10
                    Nais pardalis                8
                    Nais simplex"                6
                    Nais variabilis             10
                    Pristina                     8
                    Stylaria                     8
                    Tubificidae
                      Aulodrilus                 8
                      Limnodrilus               10

                  Hirudinea
                    Helobdella                  10

                  Turbellaria                    A
(a)  These values are for use with the biotic index scale of 0-10.
     Additional tolerance values are available in Bode (1988).
                                   C-3

-------
   Requirements and Pollution Tolerance of Trichop-
   tera. Report No.  EPA-600/4-78-063. U.S. EPA,
   Washington.
Hilsenhoff, W.L. 1982. Using a Biotic Index to Evalu-
   ate Water Quality in Streams. Technical Bulletin
   No. 132. Department of Natural Resources, Madi-
   son, Wisconsin.
Hilsenhoff, W.L. 1987.  An improved biotic  index of
   organic stream pollution. Great Lakes Entomologist
   20:31-39.
Hilsenhoff, W.L. 1988. Rapid field assessment of
   organic pollution with a family-level biotic index. J.
   N.  Am. Benthol. Soc. 7(l):65-68.
Hubbard, M.D.  and W.L. Peters.  1978. Environmental
   Requirements and Pollution Tolerance of
   Ephemeroptera. Report No. EPA-600/4-78-061. U.S.
   EPA, Washington.
Shackleford, B.  1988. Rapid Bioassessment  of Lotic
   Macroinvertebrate Communities: Biocriteria
   Development. Arkansas Department of Pollution
   Control and Ecology, Little Rock,  Arkansas.
Surdick, R.F.  and A.R. Gaufin.  1978. Environmental
   Requirements and Pollution Tolerance of Plecop-
   tera. Report No.  EPA-600/4-78-062. U.S. EPA,
   Cincinnati.
U.S. Department of Agriculture.  1985. Fisheries Sur-
   vey Handbook, Aquatic Ecosystem Inventory,
   Chapter 5 Aquatic Macroinvertebrate Surveys.
   Document No. R-4 FSH 2609.23. U.S. Department
   of Agriculture,' Forest Service, Ogden, Utah.
Weber, C.I. 1973. Biological  Field and Laboratory
   Methods for Measuring the Quality of Surface
   Waters and Effluents. Report  No. EPA-670/4-73-001.
   U.S. EPA, Cincinnati.
Winget, R.N. and F.A. Mangum. 1979. Biotic Condi-
   tion Index: Integrated Biological, Physical,  and
   Chemical Stream Parameters  for Management. U.S.
   Department of Agriculture, Forest Service,  Ogden,
   Utah.
    C.4 A PARTIAL LISTING  OF
       AGENCIES THAT  HAVE
     DEVELOPED TOLERANCE
    CLASSIFICATIONS AND/OR
            BIOTIC INDICES
Florida Department of Environmental Regulation
Illinois EPA
New York Department of Environmental Conservation
North Carolina Department of Environmental
  Management
Ohio EPA
U.S. Department of Agriculture, Forest Service,
  Intermountain Region
U.S. EPA Region V
Vermont Department of Environmental Conservation
                                                C-4

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       APPENDIX D

TOLERANCE, TROPHIC GUILDS, AND
ORIGINS OF SELECTED FISH SPECIES

-------
                                APPENDIX D
               TOLERANCE, TROPHIC GUILDS,  AND
               ORIGINS OF SELECTED FISH SPECIES
     D.1 SPECIES-LEVEL FISH
  TOLERANCE, TROPHIC, AND
    ORIGIN CLASSIFICATIONS
  Calculation of the IBI and assessment of biotic
integrity requires classification of fish species toler-
ances to an array of stressors and/or the species'
characteristic trophic guilds (Table D-l). Classifica-
tions 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 C.E. Bond (1988,
      personal communication). Those for midwestern fishes
      were taken from Karr et al. (1986) and Ohio EPA
      (1987b). Classifications for other species and regions
      can be developed using Lee et al. (1980), the regional
      fish texts listed in Section D.2, and various journal
      manuscripts, theses, and grey literature.
        Agencies contemplating the development of
      regional IBIs may wish to contact the author of this
      protocol, authors of the IBI papers cited in Section 7,
      or the State agencies now using the IBI (Section D.3)
      for further guidance.
         TABLE D-l  TOLERANCE,  TROPHIC GUILDS, AND  ORIGINS OF SELECTED
                    FISH SPECIES^'
  WILLAMETTE SPECIES

  Salmonidae
    Chinook salmon
    Cutthroat trout
    Mountain whitefish
    Rainbow trout
  Cyprinidae
    Chiselmouth
    Common  carp
    Goldfish
    Leopard dace
    Longriose dace
    Northern squawfish
    Peamouth
    Redside shiner
    Speckled dace
  Catostomidae
    Largescale sucker
    Mountain sucker
                                     Trophic Level
piscivore
insectivore
insectivore
insectivore

herbivore
omnivore
omnivore
insectivore
insectivore
piscivore
insectivore
insectivore
insectivore

omnivore
herbivore
                   Tolerance
intolerant
intolerant
intolerant
intolerant

intermediate
tolerant
tolerant
intermediate
intermediate
tolerant
intermediate
intermediate
intermediate

tolerant
intermediate
                 Origin
native
native
native
native

native
exotic
exotic
native
native
native
native
native
native

native
native
  (a)Not  necessarily the  final  designations; designations may vary  for
       different regions.
                                       D-l

-------
                            TABLE D-l  (Cont.)
Ictaluridae
  Brown bullhead
  Yellow bullhead
Percopsidae
  Sand roller
Gasterosteidae
  Threespine stickleback
Centrarchidae
  Bluegill
  Largemouth bass
  Smallmouth bass
  White crappie
Percidae
  Yellow perch
Cottidae
  Paiute sculpin
  Prickly sculpin
  Reticulate sculpin
  Torrent sculpin
Trophic Level

insectivore
insectivore

insectivore

insectivore

insectivore
piscivore
piscivore
insectivore

insectivore

insectivore
insectivore
insectivore
insectivore
 Tolerance

tolerant
tolerant

intermediate

intermediate

tolerant
tolerant
intermediate
tolerant

intermediate

intolerant
intermediate
tolerant
intolerant
Origin

exo tic
exotic

native

native

exotic
exotic
exotic
exo t i c

exotic

native
native
native
native
MIDWEST SPECIES

Petromyzontidae
  Silver lamprey
  Northern brook lamprey
  Mountain brook lamprey
  Ohio lamprey
  Least brook lamprey
  Sea lamprey
Polyodontidae
  Paddlefish
Acipenseridae
  Lake sturgeon
  Shovelnose sturgeon
Lepisosteidae
  Alligator gar
  Shortnose gar
  Spotted gar
  Longnose gar
Amiidae
  Bowfin
Hiodontidae
  Goldeye
  Mooneye
Clupeidae
  Skipjack herring
  Alewife
  Gizzard shad
  Threadfin shad
Salmonidae
  Brown trout
piscivore
filterer
filterer
piscivore
filterer
piscivore

filterer

invertivore
insectivore

piscivore
piscivore
piscivore
piscivore

piscivore

insectivore
insectivore

piscivore
invertivore
omnivore
omnivore

insectivore
intermediate
intolerant
intolerant
intolerant
intermediate
intermediate

intolerant

intermediate
intermediate

intermediate
intermediate
intermediate
intermediate

intermediate

intolerant
intolerant

intermediate
intermediate
intermediate
intermediate

intermediate
native
native
native
native
native
exotic

native

native
native

native
native
native
native

native

native
native

native
exotic
native
native

exotic
                                    D-2

-------
TABLE D-l (Cont.)

Rainbow trout
Brook trout
Lake trout
Coho salmon
Chinook salmon
Lake herring
Lake whitefish
Osmeridae
Rainbow smelt
Umbridae
Central mudminnow
Esocidae
Grass pickerel
Chain pickerel
Northern pike
Muskellunge
Cyprinidae
Common carp
Goldfish
Golden shiner
Horneyhead 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
Rosyface shiner
Redfin shiner
Rosefin shiner
Striped shiner
Common shiner
River shiner
Spottail shiner
Blackchin shiner
Bigeye shiner
Steelcolor shiner
Spotfin shiner
Bigmouth shiner
Sand shiner
Trophic Level
insectivore
insectivore
piscivore
piscivore
piscivore
piscivore
piscivore

' invertivore

insectivore

piscivore
piscivore
piscivore
piscivore

omnivore
omnivore
omnivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
generalist
insectivore
generalist
insectivore
insectivore
herbivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
Tolerance
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate
intermediate

intermediate

tolerant

intermediate
intermediate
intermediate
intermediate

tolerant
tolerant
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
Origin
exotic
native
native
exotic
exotic
native
native

exotic

native

native
native
native
native

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
D-3

-------
TABLE D-l (Cont.)

Mimic shiner
Ghost shiner
Blacknose shiner
Pugnose shiner
Silver jaw minnow
Mississippi silvery
minnow
Bullhead minnow .
Bluntnose minnow
Fathead minnow
Central stoneroller
Popeye shiner
Grass carp
Red shiner
Brassy minnow
Central silvery minnow
Catostomidae
Blue sucker
Bigmouth buffalo
Black buffalo
Smallmouth buffalo
Quillback
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 mad torn
Slender mad torn
Freckled mad torn
Northern mad torn
Scioto mad torn
Trophic Level
insectivore
insectivore
insectivore
insectivore
insectivore

herbivore
omnivore
omnivore
omnivore
herbivore
insectivore
herbivore
omnivore
omnivore
herbivore

insectivore
insectivore
insectivore
insectivore
oranivore
omnivore
omnivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
invertivore
insectivore
omnivore
insectivore
insectivore
insectivore
insectivore

piscivore
generalist
insectivore
insectivore
insectivore
insectivore
piscivore
insectivore
insectivore
insectivore
insectivore
insectivore
insectivore
Tolerance
intolerant
intermediate
intolerant
intolerant
intermediate

intermediate
intermediate
tolerant
tolerant
intermediate
intolerant
intermediate
intermediate
intermediate
intolerant

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
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
native
D-4

-------
                           TABLE D-l   (Cont.)
  Brindled madtorn
  Tadpole madtorn
Anguillidae
  American eel
Cyprinodontidae
  Western banded
    killifish
  Eastern banded
    killifish
  Blackstripe topminnow
Poeciliidae
  Mosquitofish
Gadidae
  Burbot
Percopsidae
  Trout-perch
Aphredoderidae
  Pirate perch
Atherinidae
  Brook silverside
Percichthyidae
  White bass
  Striped bass
  White perch
  Yellow bass
Centrarchidae
  White crappie
  Black crappie
  Rock bass
  Smallmouth bass
  Spotted bass
  Largemouth bass
  Warmouth
  Green Sunfish
  Bluegill
  Orangespotted sunfish
  Longear sunfish
  Redear sunfish
  Pumpkinseed
Percidae
  Sauger
  Walleye
  Yellow perch
  Dusky darter
  Blackside darter
  Longhead darter
  Slenderhead darter
  River darter
  Channel darter
  Gilt darter
  Logperch
Trophic Level

insectivore
insectivore

piscivore
insectivore

insectivore
insectivore

insectivore

piscivore

insectivore

insectivore

insectivore

piscivore
piscivore
piscivore
piscivore

invertivore
invertivore
piscivore
piscivore
piscivore
piscivore
invertivore
invertivore
insectivore
insectivore
insectivore
insectivore
insectivore

piscivore
piscivore
piscivore
insectivore
insectivore
insectivore
insectivore
insectivore
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
intolerant
intermediate
intolerant
intolerant
intermediate
Origin

native
native

native
native

native
native

exotic

native

native

native

native

native
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
                                   D-5

-------
TABLE D-l (Cont.)

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
Sciaenidae
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

invertivore

insectivore
insectivore
insectivore
insectivore

insectivore
Tolerance
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
D-6

-------
   D.2 SELECTED REFERENCES
     FOR DETERMINING FISH
      TOLERANCE, TROPHIC,
  REPRODUCTIVE,  AND ORIGIN
          CLASSIFICATIONS
ALABAMA
Smith-Vaniz, W.F. 1968. Freshwater Fishes of
   Alabama. Auburn University Agricultural Experi-
   ment Station, Auburn, Alabama. 211 pp.

ALASKA
McPhail, J.D. and C.C. Lindsey.  1970. Freshwater
   Fishes of Northwestern Canada and Alaska. Bulle-
   tin No.  173.  Fisheries Research Board of Canada.
   381 pp.

Morrow, J.E. 1980. The Freshwater Fishes of Alaska.
   Alaska Northwest Publishing Company, Anchorage,
   Alaska.  300  pp.

ARIZONA
Minckley, W.L. 1973. Fishes of Arizona. Arizona
   Game and Fish Department, Phoenix, Arizona.
   293 pp.

ARKANSAS
Buchanan,  T.M. 1973. Key to the Fishes  of Arkansas.
   Arkansas Game and Fish Commission Little Rock,
   Arkansas. 68 pp.,  198 maps.

CALIFORNIA
Moyle, P.B. 1976. Inland Fishes of California. Univer-
   sity of California Press, Berkeley, California.
   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 Divi-
   sion, Denver, Colorado. 77 pp.

CONNECTICUT
Whitworth, W.R., PL. Berrien, and W.T. Keller.
   1968. Freshwater Fishes of Connecticut. Bulletin
   No. 101. State Geological and Natural History Sur-
   vey of Connecticut. 134 pp.
DELAWARE
Lee, D.S., S.P.  Platania, C.R. Gilbert, R. Franz, and
   A. Norden. In press. A Revised List of the Fresh-
   water Fishes of Maryland and Delaware. Proceed-
   ings of the Southeastern Fishes Council.

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.R., G.H. Burgess, and R.W. Yerger. In
   preparation.  The Freshwater Fishes of Florida.

GEORGIA
Dahlberg, M.D., and D.C. Scott.  1971. The Freshwater
   Fishes of Georgia. Bulletin of the Georgia
   Academy of Science 29:1-64.

IDAHO
Simpson, J.C. and R.L. Wallace.  1978. Fishes of
   Idaho. The University  of Idaho Press, Moscow,
   Idaho. 237 pp.

ILLINOIS
Forbes, S.A. and R.E. Richardson. 1908. The Fishes
   of Illinois. Illinois State Laboratory of Natural His-
   tory. 357 pp., plus separate atlas containing 102
   maps.

Forbes, S.A. and R.E. Richardson. 1920. The Fishes
   of Illinois. Second edition. Illinois Natural History
   Survey. 357 pp.

Smith, P.W. 1979. The Fishes of Illinois. Illinois  State
   Natural History Survey, University of Illinois  Press,
   Urbana, Illinois. 314 pp.

INDIANA
Gerking, S.D. 1945. The  distribution  of the fishes of
   Indiana. Investigation of Indiana Lakes and Streams
   3:1-137.

IOWA
Harlan, J.R. and E.B. Speaker. 1951. Iowa Fish and
   Fishing. State Conservation Commission, State of
   Idaho. 237 pp.

KANSAS
Cross, F.B. 1967. Handbook of Fishes of Kansas. Pub-
   lic Education Series No. 3. University of Kansas
   Museum of  Natural History. 189 pp.
                                                D-7

-------
KENTUCKY
Burr, B.M. In press. A distribution checklist of the
   fishes of Kentucky. Brimleyana No. 3.

Clay, W.M. 1975. The Fishes of Kentucky. Kentucky
   Department of Fish and Wildlife Resources, Frank-
   ford, Kentucky. 416 pp.

LOUISIANA
Douglas, N.H.  1974. Freshwater Fishes of Louisiana.
   Claitors Publishing Division, Baton Rouge, Loui-
   siana. 443 pp.

MAINE
Everhart, W.H.  1966. Fishes of Maine. Third edition.
   Maine Department of Inland Fisheries  and Game,
   Augusta, Maine. 96  pp.

MARYLAND
Lee, D.S.,  S.P.  Platania, C.R.  Gilbert, R. Franz, and
   A. Norden. In press. A Revised List of the Fresh-
   water Fishes of Maryland and Delaware. Proceed-
   ings of the Southeastern Fishes Council.

MASSACHUSETTS
Mugford, PS. 1969.  Illustrated Manual of Mas-
   sachusetts Freshwater Fish.  Massachusetts Division
   of Fish and Game, Boston,  Massachusetts. 127 pp.

MICHIGAN
                                    t
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, Michigan.  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. Underbill. 1974. Northern Fishes,
   with  Special  Reference  to the Upper Missippi Val-
   ley. University of Minnesota Press, Minneapolis,
   Minnesota. 414 pp.

Phillips, G.L. and J.C. Underbill.  1971. Distribution
   and variation  of the Catostomidae  of Minnesota.
   Occasional Papers of the Bell Museum of Natural
   History  10:1-45.
Underbill, 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.

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. Missis-
   sippi Game and Fish Commission, pp. 6-22.

Cook,  F.A. 1959. Freshwater Fishes  in Mississippi.
   Mississippi Game and  Fish Commission, Jackson,
   Mississippi. 239  pp.

MISSOURI
Pflieger, W.L. 1971. A distributional 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, Missouri.
   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. Morris, and  L.  Witt. 1972. The Fishes of
   Nebraska.  Nebraska Game and Parks Commission,
   Lincoln, Nebraska. 98 pp.

NEVADA
LaRivers, I.  1962.  Fish and Fisheries of Nevada.
   Nevada State Fish and Game Commission, Carson
   City, Nevada.  782 pp.

NEW HAMPSHIRE
Scarola, J.F.  1973. Freshwater Fishes of New Hamp-
   shire. New Hampshire Fish  and Game Department,
   Concord, New Hampshire. 131 pp.

NEW JERSEY
Stiles,  E.W. 1978. Vertebrates of New Jersey. Edmund
   W. Stiles Publishers, Somerset, New Jersey.
   148 pp.
                                                   D-8

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NEW MEXICO
Koster, W.J.  1957. Guide to the Fishes of New Mex-
   ico. University of New Mexico Press, Albuquer-
   que, New Mexico. 116 pp.

NEW YORK
Greeley, J.R.  1927-1940. Watershed survey reports on
   fishes of New York rivers, published as supple-
   ments to the 16th through 29th Annual Reports of
   the New York State Conservation Department,
   Albany, New York.

Smith, C.L.  In preparation. Inland Fishes of New
   York.

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.  In preparation. The Freshwater
   Fishes of North Carolina.

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, Ohio. 683 pp.

OKLAHOMA
Miller, R.J. and H.W. Robinson. 1973. The  Fishes of
   Oklahoma.  Oklahoma State University Press, Still-
   water, Oklahoma. 246 pp.

OREGON
Bond, C.E. 1973. Keys to Oregon freshwater fishes.
   Technical Bulletin 58:1-42.  Oregon State University
   Agricultural Experimental Station, Corvallis,
   Oregon.

PENNSYLVANIA
Cooper, E.L. In preparation. Fishes of Pennsylvania.

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. Publication No. 119. Miscellaneous Publi-
   cations of the Museum of Zoology, University of
   Michigan. 131 pp.

TENNESSEE
Etnier, DA. and W.C. Starnes. In preparation. The
   Fishes of Tennessee.

Kuhne, E.R. 1939.  A Guide to the Fishes of Tennes-
   see and the  Mid-South. Tennessee Department of
   Conservation, Nashville, Tennessee. 124 pp.

TEXAS
Hubbs, C. 1972. A checklist of Texas  freshwater
   fishes. Texas Parks and Wildlife Department Tech-
   nical Service 11:1-11.

Knapp, FT. 1953.  Fishes Found in the Fresh Waters
   of Texas. Ragland Studio and Lithograph Printing
   Company, Brunswick, Georgia. 166 pp.

UTAH
Sigler, W.F. and R.R.  Miller. 1963. Fishes of Utah.
   Utah Game  and Fish Department, Salt Lake City,
   Utah.  203 pp.

VERMONT
MacMartin, J.M.  1962. Vermont  stream  survey
   1952-1960. Vermont  Fish and Game Department,
   Montpelier,  Vermont. 107  pp.

VIRGINIA
Jenkins, R.E.,  N.M. Burkhead, and D.J. Jenkins. In
   preparation. The Freshwater Fishes of Virginia.

WASHINGTON
Wydoski, R.S.  and R.R. Whitney. 1979. Inland  Fishes
   of Washington. University of Washington Press.
   220 pp.

WEST VIRGINIA
Denoncourt, R.F.,  B.C. 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.
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WISCONSIN
Becker, G.C.  1983. Fishes of Wisconsin. University of
   Wisconsin Press, Madison, Wisconsin.  1052 pp.

WYOMING
Baxter, G.T. and J.R. Simon.  1970. Wyoming Fishes.
   Bulletin No.  4. Wyoming Game and Fish Depart-
   ment. Cheyenne, Wyoming. 168 pp.

CANADA
McPhail, J.D. and  C.C. Lindsey.  1970. Freshwater
   Fishes of Northwestern Canada and Alaska. Bulle-
   tin No. 173. Fisheries Research Board of Canada.
   381  pp.

Scott, W.B. and E.J. Grossman. 1973. Bulletin No.
   1984. Freshwater Fishes of Canada. Fisheries
   Research Board  of 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 Publica-
   tion 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 British Columbia. Fourth
   edition.  Handbook No. 5. British Columbia Provin-
   cial  Museum. 192 pp.

Hart, J.L. 1973. Pacific Fishes. Second edition. Bulle-
   tin No. 180. Fisheries Research Board of Canada.
   740  pp.

MANITOBA
Fedoruk, A.N.  1969. Checklist and Key of the Fresh-
   water Fishes of  Manitoba. Manitoba Department of
   Mines and Natural Resources, Canada Land Inven-
   tory Project. 98 pp.

Hinks, D. 1943. The Fishes of Manitoba. Manitoba
   Department of Mines and Natural Resources.
   102 pp.
NEW BRUNSWICK
Gorham, S.W.  1970. Distributional Checklist of the
   Fishes of New Brunswick. Saint John, New Brun-
   swick. 32 pp.

Scott, W.B. and E.J. Grossman.  1959. The Freshwater
   Fishes of New Brunswick. A  checklist with dis-
   tributional 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/l-l. Environmental-Social  Committee
   Northern Pipelines, Task Force on  Northern
   Development. 122 pp;

NOVA SCOTIA
Gilhen, J. 1974. The Fishes of Nova Scotia's Lakes
   and Streams. Nova Scotia Museum, Halifax. 49 pp.

Livingstone,  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. Grossman. 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 Quebec.  First English
   edition. Quebec Department of Game and Fisher-
   ies. 189 pp.

Masse G., et J. Mongeau. 1974.  Repartition Geograp-
   hique des Poissons, leur abondance relative et
   bathymetric  de la region du Lac Saint-Pierre. Ser-
   vice de lAmenagement 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.
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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 inventaires 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 cornmer-
   ciale, les ensemencements, les frayeres, la contami-
   nation par le  mercure et les PCB. Service de
   I'Amegagement de la Faune, Ministere du
   Tourisme, de la Chasse et de la Peche,  Quebec.
   286 pp.

SASKATCHEWAN
Symington, D.F. 1959.  The Fish of Saskatchewan.
   Conservation Bulletin No. 7. Saskatchewan Depart-
   ment of Natural Resources. 25 pp.

YUKON TERRITORY
Bryan, I.E.  1973. The influence of pipeline develop-
   ment 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 Pipe-
   lines, 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 Commis-
   sion, Ann Arbor,  Michigan.

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, Minnesota.  267 pp.

Hocutt, C.H. and E.O. Wiley. 1986.  The Zoogeogra-
   phy of North American Freshwater Fishes. John
   Wiley and Sons, New York.

Hubbs, C.L.  and K.F. Lagler. 1947.  Fishes of the
   Great Lakes Region. The Cranbrook  Press, Bloom-
   field Hills, Michigan. 186 pp.

Jenkins, R.E., E.A.  Lachner, and F.J. Schwartz.  1972.
   Fishes of the 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 Insti-
   tute and State University, Blacksburg, Virginia.
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 Caro-
   lina Museum of Natural History, Raleigh, North
   Carolina.

Metcalf, A.L.  1966.  Fishes of  the Kansas River sys-
   tem in relation to zoogeography of the Great
   Plains. Publication of the Museum of Natural His-
   tory, University of Kansas 17(3):23-189.

Miller, R.R. 1948. The Cyprinodont fishes of the
   Death Valley system of eastern California and south-
   western Nevada. Miscellaneous Publication of the
   Museum of Zoology, University of Michigan
   68:1-55.

Miller, R.R. 1959. Origin and  Affinities of the Fresh-
   water Fish Fauna of Western North America. Zoo-
   geography Publication Number 51. American
   Association  for the Advancement of Science,
   Washington, D.C.

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 Commissions,
   pp. 10-27.

Soltz, D.L. and R.J. Naiman.  1978. The natural his-
   tory of native fishes in the Death Valley system.
   Natural History Museum of Los Angeles County.
   Science Series 30:1-76.
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