&EPA
United States
Environmental Protection
Agency
A Review of Biological Assessment Tools
and Biocriteria for Rivers and Streams in
New England States
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EPA/600/R-04/168
October 2004
A Review of Biological Assessment Tools and Biocriteria for Streams and
Rivers in New England States
Alicia D. Shelton
SoBran, Inc.
Karen A. Blocksom
National Exposure Research Laboratory
U.S. Environmental Protection Agency
National Exposure Research Laboratory
26 West Martin Luther King Drive
Cincinnati, OH 45268
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NOTICE
The research described in this document has been funded by the United States
Environmental Protection Agency under contract 68D01048 to SoBran, Inc. It has been
subjected to Agency peer and administrative review and approved for publication as an EPA
document.
Mention of trade names or commercial products does not constitute endorsement or
recommendation for use.
The correct citation for this document is:
Shelton, A.D., and K.A. Blocksom. 2004. A Review of Biological Assessment Tools and
Biocriteria for Streams and Rivers in New England States. EPA/600/R-04/168. U.S.
Environmental Protection Agency, Cincinnati, Ohio.
Cover photos by (clockwise from upper left): New Hampshire DES Biomonitoring Program;
Hilary Snook, USEPA Region 1; Richard Levey, Vermont DEC; NHDES Biomonitoring
Program.
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ACKNOWLEDGMENTS
This report relied on the generous assistance of state personnel in providing materials on
bioassessment tools and biocriteria and reviewing draft chapters for each state. Thanks go to
Ernest Pizzuto, Jr. of Connecticut DEP, Susan P. Davies of Maine DEP, Arthur S. Johnson of
Massachusetts DEP, David Neils of New Hampshire DES, Connie Carey of Rhode Island DEM,
and Doug Burnham, Richard Levey, and Richard Langdon of Vermont DEC.
Peter Nolan of U.S. EPA Region 1, Wayne Davis of the U.S. EPA Office of Environmental
Information, and Bradley Autrey of the U.S. EPA Office of Research and Development all
provided valuable comments and suggestions on a draft of this document. Eric O'Neal of
SoBran, Inc. provided assistance in creating some of the maps in this report. Michael T. Barbour
of Tetra Tech, Inc. provided the suggestion for the general layout of the document.
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TABLE OF CONTENTS
LIST OF FIGURES AND TABLES vii
ACRONYMS AND COMMON TERMS x
1 INTRODUCTION 1-1
1.1 Purpose of the Document 1-1
1.2 Rational for Bioassessment Programs 1-1
1.2.1 Designated Uses 1-3
1.2.2 Water Quality Criteria for Aquatic Life Use 1-3
1.2.3 Anti-degradation Policies 1-4
1.2.4 Guidance Documents 1-4
1.2.5 Biological Monitoring Programs 1-5
1.2.6 Bioindicator Organisms 1-5
1.2.7 305 (b) Report and 303 (d) List 1-6
1.3 Literature Cited 1-7
2 CONNECTICUT 2-1
2.1 Introduction 2-1
2.2 Key Elements of the Biological Assessment Approach 2-1
2.2.1 Index Period and/or Temporal Conditions 2-1
2.2.2 Monitoring Program Survey Approach 2-3
2.2.3 Indicator Assemblages 2-3
2.2.4 Reference Condition 2-4
2.3 Field and Laboratory Protocols 2-5
2.3.1 Macroinvertebrate Protocols 2-5
2.3.1.1 Field Methods 2-5
2.3.1.2 Laboratory Methods 2-5
2.3.2 Periphyton Protocols 2-5
2.3.2.1 Field Methods 2-5
2.3.2.1.1 Quantitative Periphyton Sampling 2-5
2.3.2.1.2 Rapid Periphyton Survey 2-6
2.3.2.2 Laboratory Methods 2-6
2.3.2.2.1 Chlorophyll a 2-6
2.3.2.2.2 Algal Identification and Density 2-7
2.3.2.2.3 Biomass and Biovolume Determination 2-7
2.3.3 Fish Protocol 2-7
2.4 Data Management/Quality 2-7
2.5 Analysis of Biological Data 2-8
2.5.1 Macroinvertebrate Data 2-8
2.5.2 Periphyton Data 2-9
2.5.3 Fish Data 2-9
2.5.4 Summary: Determining ALU Support 2-10
2.6 Literature Cited 2-11
in
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2.7 Resources 2-12
3 MAINE 3-1
3.1 Introduction 3-1
3.2 Key Elements of the Biological Assessment Approach 3-3
3.2.1 Index Period and/or Temporal Conditions 3-3
3.2.2 Monitoring Program Survey Approach/Natural Classification of Water Bodies... 3-4
3.2.3 Indicator Assemblage 3-4
3.2.4 Reference Condition (Establishing a priori Groups) 3-4
3.3 Field and Laboratory Protocols 3-5
3.3.1 Macroinvertebrate Protocols 3-5
3.3.1.1 Field Methods 3-5
3.3.1.2 Laboratory Methods 3-6
3.3.1.2.1 Subsampling and Identification 3-6
3.3.1.2.2 Chironomidae Identification and Subsampling 3-7
3.4 Data Management/Quality 3-7
3.5 Analysis of Biological Data 3-7
3.6 Literature Cited 3-17
3.7 Resources 3-20
4 MASSACHUSETTS 4-1
4.1 Introduction 4-1
4.2 Key Elements of the Biological Assessment Approach 4-2
4.2.1 Index Period and/or Temporal Conditions 4-2
4.2.2 Monitoring Program Survey Approach 4-2
4.2.3 Natural Classification of Water Bodies 4-3
4.2.4 Indicator Assemblages 4-3
4.2.5 Reference Condition 4-3
4.3 Field and Laboratory Protocols 4-4
4.3.1 Macroinvertebrate Protocols 4-4
4.3.1.1 Field Methods 4-4
4.3.1.1.1 Kick Sampling 4-6
4.3.1.1.2 Rock Basket Sampling 4-6
4.3.1.1.3 Hester-Dendy Multi-plate Sampling 4-6
4.3.1.2 Laboratory Methods 4-7
4.3.1.2.1 Processing of Kick Net and Rock Basket Samples 4-7
4.3.1.2.2 Processing of Hester-Dendy Multi-plate Samples 4-7
4.3.1.2.3 Taxonomic Identification 4-8
4.3.1.2.4 Oligochaeta and Chironomidae Identification 4-8
4.3.2 Periphyton Protocols 4-8
4.3.2.1 Field Methods 4-8
4.3.2.1.1 Algal Abundance and Identification 4-8
4.3.2.1.2 Biomass 4-9
4.3.2.1.3 Chlorophyll a 4-9
4.3.2.1.4 Percent coverage calculation 4-9
IV
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4.3.2.1.5 Biomass Determination 4-10
4.3.2.1.6 Chlorophyll a Determination 4-11
4.3.3 Fish Protocols 4-11
4.4 Data Management/Quality 4-12
4.5 Analysis of Biological Data 4-12
4.5.1 Macroinvertebrate Data 4-12
4.5.2 Algal Data 4-15
4.5.3 Fish Data 4-15
4.5.4 Summary: Determining ALU Support 4-15
4.6 Literature Cited 4-17
4.6.1 Resources 4-20
5 NEW HAMPSHIRE 5-1
5.1 Introduction 5-1
5.2 Key Elements of the Biological Assessment Approach 5-3
5.2.1 Index Period and/or Temporal Conditions 5-3
5.2.2 Monitoring Program Survey Approach 5-3
5.2.3 Natural Classification of Water Bodies 5-3
5.2.4 Indicator Assemblages 5-3
5.2.5 Reference Condition 5-3
5.3 Field and Laboratory Protocols 5-5
5.3.1 Macroinvertebrates Protocols 5-5
5.3.1.1 Field Methods 5-5
5.3.1.2 Laboratory Methods 5-5
5.3.2 Fish Protocol 5-6
5.4 Data Management/Quality 5-6
5.5 Analysis of Biological Data 5-6
5.5.1 Macroinvertebrate Data 5-6
5.5.2 Fish Data 5-7
5.6 Summary: Determining ALU Support 5-8
5.7 Literature Cited 5-9
5.8 Resources 5-10
6 RHODE ISLAND 6-1
6.1 Introduction 6-1
6.2 Key Elements of the Biological Assessment Approach 6-2
6.2.1 Index Period and/or Temporal Conditions 6-2
6.2.2 Monitoring Program Survey Approach 6-2
6.2.3 Natural Classification of Water Bodies 6-2
6.2.4 Indicator Assemblages 6-2
6.2.5 Reference Condition 6-2
6.3 Field and Laboratory Protocols 6-3
6.3.1 Macroinvertebrate Protocols 6-3
6.3.1.1 Field Methods 6-3
6.3.1.2 Laboratory Methods 6-4
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6.4 Data Management/Quality 6-4
6.5 Analysis of Biological Data 6-5
6.5.1 Macroinvertebrate Data 6-5
6.6 Summary: Determining ALU Support 6-6
6.7 Literature Cited 6-9
6.8 Resources 6-9
7 VERMONT 7-1
7.1 Introduction 7-1
7.2 Key Elements of the Biological Assessment Approach 7-3
7.2.1 Index Period and/or Temporal Conditions 7-3
7.2.2 Monitoring Program Survey Approach 7-3
7.2.3 Natural Classification of Water Bodies 7-3
7.2.4 Indicator Assemblages 7-3
7.2.5 Reference condition 7-3
7.3 Field and Laboratory Protocols 7-5
7.3.1 Macroinvertebrate Protocols 7-5
7.3.1.1 Field Methods 7-5
7.3.1.2 Laboratory Methods 7-5
7.3.2 Fish Protocols 7-6
7.3.2.1 Field Methods 7-6
7.4 Data Management/Quality 7-6
7.5 Analysis of Biological Data 7-7
7.5.1 Macroinvertebrate Data 7-7
7.5.2 Fish Data 7-9
7.6 Summary: Determining ALU Support 7-10
7.7 Literature Cited 7-14
7.8 Resources 7-15
8 SUMMARY 8-1
8.1 Comparison Across States 8-1
8.2 Literature Cited 8-8
APPENDIX A: PROCESS AND CRITERIA FOR THE ASSIGNMENT OF
BIOLOGIST'S CLASSIFICATION A-l
VI
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LIST OF FIGURES AND TABLES
FIGURES
Figure 2-1. Major Connecticut basins sampled for the biological monitoring program using the
rotating basin strategy 2-4
Figure 3-1. Maine's narrative aquatic life standards with the human disturbance and biological
condition gradients (Taken from Courtemanch 2003) 3-3
Figure 3-2. Map of basins sampled by MDEP (2002) 3-5
Figure 3-3. Maine tiered uses based on measurable ecological values (taken from Courtemanch
2003) 3-9
Figure 3-4. Process of calculating model variables and association values using linear
discriminant models (taken from MDEP 2003) 3-18
Figure 3-5. Process for determining attainment class using association values (modified from
MDEP 2003) 3-19
Figure 4-1. Massachusetts 5-Year Basin Rotation Strategy (taken from the Massachusetts
Department of Environmental Protection website
www.mass.gov/dep/brp/wm/files/cyclemap6.jpg) 4-4
Figure 4-2. Level III and Level IV Ecoregions of Massachusetts (taken from Griffith et al. 1994,
http://www.epa.gov/wed/pages/ecoregions/mactri eco.htm) 4-5
Figure 5-1. Major New Hampshire basins and the northern and southern bioregion boundaries
used for macroinvertebrate sampling (indicated by the red line) 5-4
Figure 6-1. Level IV Omernik subecoregions and reference streams used in RI OEM's biological
monitoring program 6-3
TABLES
Table 1 -1. Contact information for bioassessment programs in New England states 1-2
Table 2-1. Connecticut water quality standard classes 2-2
Table 2-2. Metrics and scoring ranges used in RBP III determinations of the level of biological
impact based on benthic macroinvertebrates (based on Plafkin etal. (1989)) 2-8
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Table 2-3. Aquatic life use support categories and the criteria used for making decisions (taken
from Table 2 in CT DEP 2002a) 2-10
Table 3-1. Water quality classification system for rivers and streams in Maine. (M.R.S.A. Title
38 Article 4-A §464-465) 3-2
Table 3-2. Methods for the calculation of variables and measures of community structure used in
linear discriminant models (from Davies and Tsomides, 2002) 3-10
Table 3-3. Coefficients for the First Stage Model (from MDEP 2003) 3-14
Table 3-4. Coefficients for the Final Classification Models (AA/A, B, and C) (MDEP 2003).3-14
Table 4-1. Massachusetts attainment classes with management strategy and narrative biologic
and habitat criteria as stated in 314 CMR 4.00 (2000) 4-2
Table 4-2. Methods for the calculations of metrics and scoring ranges used in RBP II
determinations of level of biological impact (Plafkin 1989; Nuzzo 2003) 4-12
Table 4-3. Methods for the calculations of metrics and scoring ranges used in RBP III
determinations of level of biological impact (Plafkin 1989; Nuzzo 2003) 4-14
Table 4-4. Biological, lexicological, and chemical parameters that are used collectively to
determine ALUS. Attainment is assigned based on a "weight of evidence" evaluation. (MA DEP
2003) (Numerical criteria for dissolved oxygen, pH, and temperature can be found in 314 CMR
4.00 (MA DEP 2000). MA DEP uses the recommended limits published by EPA pursuant to
Section 3 04(a) of the Federal Act for Toxic Pollutant Criteria) 4-16
Table 5-1. NH DES water quality classes and the defined designated uses for each class.
Dissolved oxygen exceedance values for aquatic life criteria are also listed (NH DES 1999 ).. 5-2
Table 5-2. Metrics and scoring for the New Hampshire B-IBI 5-8
Table 6-1. Metrics used by the Rhode Island Biomonitoring program and the methods for the
calculation of metrics and their scoring ranges based on the RBP III (Plafkin et al. 1989, RI
DEM 2002a, RI DEM 2002b) 6-5
Table 6-2. Percent comparability evaluation for macroinvertebrate bioassessment scores used by
the State of Rhode Island 6-7
Table 6-3. Biological, physical and chemical criteria used to determine aquatic life use (modified
from RI DEM 2000) 6-7
Vlll
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Table 7-1. Biological ALUS management classes and associated narrative biological criteria for
rivers and streams in Vermont 7-2
Table 7-2. VT DEC macroinvertebrate metrics and methods used to calculate each of the
metrics 7-7
Table 7-3. Macroinvertebrate assemblage biocriteria thresholds for the macroinvertebrate
community stream categories, and associated WQ classes of Vermont (VT DEC 2004) 7-11
Table 7-4. The six metrics used in scoring the fish assemblage for the CWIBI. These streams
must naturally support two to four native species (VT DEC 2004) 7-12
Table 7-5. The nine metrics used in scoring cold and warm water sites for the MWTBI. These
streams must naturally support more than four native fish species (VT DEC 2004) 7-12
Table 7-6. All possible scores for the CWIBI and MWIBI that correspond to the VT WQS
classification scheme (VT DEC 2004) 7-14
Table 8-1. Comparisons of the key components of state bioassessment programs 8-2
Table 8-2. Comparison of the macroinvertebrate metrics used by states in the New England
Region. Color shading indicates equivalent metrics across states 8-6
IX
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ACRONYMS AND COMMON TERMS
305(b) Report: Clean Water Act (CWA) section 305(b) requires each state to submit an
assessment report biennially to U.S. EPA on the quality of surface and ground water resources.
The EPA then compiles the data from state 305(b) reports and submits a National Water Quality
Report to Congress
303(d) List: The section of the Clean Water Act that requires each state to identify waters that
are impaired according to water quality standards. Placement of water bodies on this list requires
the preparation of Total Maximum Daily Loads (TMDLs) that will aid in the cleanup of the
impacted waters.
7Q10: The lowest consecutive 7-day mean stream flow that occurs during a 10-year period
30Q5: The lowest consecutive 30-day mean stream flow that occurs during a 5-year period.
ABN: Ambient Biomonitoring Network [Vermont]
AFDM: Ash Free Dry Mass
ALU: Aquatic Life Use
ALUS: Aquatic Life Use Support
BASS: Biomonitoring and Aquatic Studies Section [Vermont]
B-IBI: Benthic Index of Biotic Integrity. [New Hampshire]
BPJ: Best Professional Judgment
CALM: Consolidated Assessment and Listing Methodology
CT DEP: Connecticut Department of Environmental Protection
CWA: Clean Water Act
CWIBI: Cold Water Index of Biotic Integrity [Vermont]
DO: Dissolved oxygen
EDAS: Ecological Data Assessment System: A database system developed by Tetra Tech, Inc.
that uploads ecological data to STORET for archival and has the capability to analyze, manage
and store data and calculate metrics.
EMAP: Environmental Monitoring and Assessment Program
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EPT: Insect orders of Ephemeroptera, Plecoptera, Trichoptera, considered sensitive benthic
orders.
FBI: Family Biotic Index
GIS: Geographic Information Systems
GWHI: Ground Water Hazard Inventory
HBI: Hilsenhoff Biotic Index
HDG: Human Disturbance Gradient [New Hampshire]
HUC: Hydrologic Unit Code
IBI: Index of Biotic Integrity
MDEP: Maine Department of Environmental Protection
NA: non-attainment
MA DWM: Massachusetts Division of Watershed Management
MA DEP: Massachusetts Department of Environmental Protection
MHG: Medium High Gradient Streams [Vermont]
NH DES: New Hampshire Department of Environmental Services
MWIBI: Mixed Water Index of Biotic Integrity [Vermont]
NEWS: New England Wadeable Streams Project
NHLC: New Hampshire Land Cover
NPDES: National Pollutant Discharge Elimination System
ONRW: Outstanding National Resource Waters
QAPP: Quality Assurance Project Plan
RCRA: Resource Conservation Recovery Act
RBP: Rapid Bioassessment Protocol
RI DEM: Rhode Island Department of Environmental Management
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RPS: Rapid Periphyton Survey (RBP)
SHG: Small High Gradient Streams [Vermont]
SOP: Standardized Operating Procedure
TMDL: Total Maximum Daily Load
WBS: Water Body System- Database developed by USEPA to store information to be used in
305(b) reporting and 303(d) listing for all assessed water bodies within a region
WPCA: Water Pollution Control Act
WQS: Water Quality Standards
WWMG: Warm water medium grade streams and rivers [Vermont]
VT DEC: Vermont Department of Environmental Conservation
xn
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1 INTRODUCTION
1.1 Purpose of the Document
The primary purpose of this document is to serve as a detailed description of the
biological assessment programs for wadeable streams and rivers within U.S. Environmental
Protection Agency (U.S. EPA) Region 1 states (i.e., Connecticut, Maine, Massachusetts, New
Hampshire, Rhode Island, and Vermont). Specifically, this report concentrates on the target
assemblages (e.g., benthic macroinvertebrates, periphyton, and/or fish) and the specific methods
used by each state to determine whether biocriteria set for aquatic life use (ALU) are met in
wadeable streams and rivers. The information contained in this report is critical to the eventual
use of state data in assessing water resources on a national scale because it provides the
necessary level of detail on New England state bioassessment methodologies in a single
document. In addition, this report serves as a valuable resource for other states, tribes, and
municipalities, both those developing bioassessment tools and those with existing programs.
Although every attempt has been made to represent the methods and protocols used by
each state accurately, this document is not intended to be used as a replacement for those
protocols and Standard Operating Procedures (SOPs) that are used and approved by the state
agencies. Thanks to the cooperation of state scientists, all protocols and procedures were
obtained through personal communication and via state and federal published and unpublished
documents that are referenced within this report. Each state reviewed its respective chapter for
technical accuracy and was given the opportunity to provide comments and changes prior to
completion of this report. However, we recommend referring directly to state protocols before
implementation of the described methods to ensure that the most updated and complete versions
of protocols are used. Contact details for each of the state bioassessment programs discussed are
provided in Table 1-1.
1.2 Rational for Bioassessment Programs
The modern Clean Water Act (CWA) is derived from the 1948 Federal Water Pollution
Control Act (WPCA). After the passage of the 1972 amendments, the act became commonly
known as the CWA and its goal was to "restore and maintain the chemical, physical and
biological integrity of the nation's waters so that they can 'support the protection and
propagation offish, shellfish, and wildlife and recreation in and on the water'"
(http://www.epa.gov/watertrain/cwa/). This act federally recognized the aquatic inhabitants of
water bodies and began to set water quality standards to protect these organisms. The CWA
amendments through 1987 outlined the guidelines by which states and tribes must use
bioassessment programs and develop biocriteria to ensure the adherence to water quality
standards. Specifically, Section 303(c) of the CWA requires states to have water quality
standards (WQS) that consist of three components: 1) designated uses, 2) water quality criteria to
protect those uses, and 3) an anti-degradation policy. States are required to review their
standards every three years and revise them as needed to achieve the purposes of the CWA,
including the ecological integrity objective.
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Table 1-1. Contact information for bioassessment programs in New England states.
State Program Contact Web Site
Connecticut Ernest Pizzuto, Jr.
Supervising Environmental Analyst
Connecticut Department of Environmental
Protection
Address:
79 Elm St.
Hartford, CT 06106-5127
Phone: 860-424-3715
Email: ernest.pizzuto@po.state.ct.us
http://dep.state.ct.us/
Maine
Susan P. Davies
Program Manager, Biologist III
Maine Department of Environmental Protection
Address:
SHS 17
Augusta, ME 04333
Phone: 207-287-7778
Email: susan.p.davies@maine.gov
http://www.maine.gov/dep/
Massachusetts
Arthur S. Johnson
Environmental Monitoring Coordinator
Massachusetts Department of Environmental
Protection
Address:
627 Main Street
Worcester, MA 01608
Phone: 508-767-2873
Email: arthur.johnson@state.ma.us
http ://www. state.ma.us/dep/
New David Neils
Hampshire Biomonitoring Program Coordinator
New Hampshire Department of Environmental
Services
Address:
6 Hazen Drive
Concord, NH 03302-0095
Phone: 603-271-8865
Email: dneils@des.state.nh.us
www.des.state.nh.us/
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State
Program Contact
Web Site
Rhode Island
Connie Carey
Environmental Scientist
Rhode Island Department of Environmental
Management
Address:
235 Promenade Street
Providence, RI 02908-5767
Phone: 401-222-4700 x7239
Email: ccarey@dem.state.ri.us
http ://www. state.ri .us/dem/
Vermont
Doug Burnham
Biomonitoring and Aquatic Studies Section
Chief
Vermont Department of Environmental
Conservation
Address:
103 South Main Street-ION
Waterbury, VT 05671
Phone: 802-241-3784
Email: doug.burnham@anr.state.vt.us
http ://www.anr. state, vt.us/
1.2.1 Designated Uses
As required by CWA 40 C.F.R. § 130.10, states, territories and tribes must specify
appropriate beneficial uses based on the intended use and the value of the waters, and these uses
must be achieved and protected. Designated uses may be listed as general categories (e.g.,
drinking water source, wildlife, shellfish, aquatic life, recreational, industrial), or the uses may
consist of more specific sub-categories that may target cold water versus warm water systems or
contain special uses that are meant to protect unique, sensitive, or valuable aquatic habitat (U.S.
EPA 1991). These designated uses are typically associated with a classification system (e.g.,
Class A waters, Class B waters, Class C waters) within each state's WQS that categorizes each
water body according to condition.
1.2.2 Water Quality Criteria for Aquatic Life Use
Water quality criteria are narrative or numeric descriptions of those conditions that
protect designated uses. In addition, the criteria need to be scientifically consistent with the
intended designated use and must be accurate indicators of the designated use. Although the
U.S. EPA has published guidance criteria to protect aquatic life use (U.S. EPA 2002a),
individual states are not required to follow them and may develop their own criteria. Guidance
for the development of numeric criteria are published in the CWA § 104(a)(l) and may be
modified based on the needs of the state. Currently, only narrative descriptions of criteria for
aquatic life use support are required within state WQS by the U.S. EPA. The narrative criteria
are simply descriptions of the conditions necessary for a water body to attain its designated use
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(U.S. EPA 2002b), and these definitions, along with organisms that can be used to assess
attainment, vary from state to state in U.S. EPA Region 1.
Each state in Region 1 has established its goal for protecting waters and then defined
aquatic life use (ALU). For example, New Hampshire statutes define waters achieving ALU as
those waters that "provide suitable chemical and physical conditions for supporting a balanced,
integrated and adaptive community of aquatic organisms (NH DES 1999). Aquatic life use
support (ALUS) as defined in Rhode Island WQS is "providing suitable habitat and water quality
for the protection, maintenance, and propagation of a viable community of aquatic life" (RI
DEM 2000). Section 3-01 of the Vermont Water Quality Standards states the provision to,
"establish and apply numeric biological indices to determine whether there is full support of
aquatic biota and aquatic habitat uses" and to "establish procedures that employ standard
sampling and analytical methods to characteristics of the biological integrity of the appropriate
reference conditions" (State of Vermont 2000). In Massachusetts, the ALUS criteria of the
standards 314 CMR 4.00 "must provide suitable habitat for sustaining a native, naturally diverse
community of aquatic flora and fauna" (MA DEP 2000; 2003). Massachusetts then further
designates two subclasses: Cold Water Fishery - capable of sustaining a year-round population of
cold water aquatic life; and Warm Water Fishery - waters that are not capable of sustaining a
year-round population of cold water aquatic life (MA DEP 2003). Connecticut WQS express
that "the benthic invertebrate criteria may be utilized where appropriate for assessment of the
biological integrity of surface waters. These criteria apply to the fauna of erosional or riffle
habitats in lotic waters which are not subject to tidal influences" (CT DEP 2002). Connecticut
defines biological integrity as the "ability of an aquatic ecosystem to support and maintain a
balanced, integrated, adaptive community of organisms having a species composition, diversity,
and functional organization comparable to that of the natural habitats of a region" (CT DEP
2002). In Maine, the use of benthic organisms to determine the attainment of ALU is written
directly into the standards in chapter 579 (MDEP 2003). Chapter 579 gives a detailed
description of the use of benthic organisms and the methods used to make decisions about
classification attainment (MDEP 2003). Furthermore, narrative standards in Maine Revised
Statutes Annotate 38 Public Chapter 3 Article 4-A § 464 and § 465 define the biological
narrative and numerical dissolved oxygen and bacterial standards.
1.2.3 Anti-degradation Policies
The anti-degradation policy (CWA 40 CFR §131.12) is a set of rules designed to protect
high quality waters. This policy must offer a framework of decision-making if water quality
changes occur. The U.S. EPA WQS require states to implement a three-tiered system for
addressing anti-degradation. Tier 1 requires that water quality necessary to support existing uses
is maintained and protected, Tier 2 states that in no case shall water quality decrease to a level
that would interfere with the designated use, and Tier 3 maintains and protects outstanding
national resource waters (ONRW), aiming to preserve those waters with exceptional recreational
or ecological significance (U.S. EPA www.epa.gov/waterscience/standards/about/adeg.htm).
1.2.4 Guidance Documents
To support the assessment of attainment of beneficial uses, states are responsible for
implementing a biological monitoring strategy for the design, collection and data analysis of
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biological data. The U.S. EPA has published technical documents and guidance documents to
offer support for the development of state biomonitoring programs for the assessment of water
quality for ALUS. The most current documents include the Guidance for 2004 Assessment
Listing and Reporting Requirements Pursuant to Sections 303(d) and 305(b) of the Clean Water
Act: TMDL-01-03 (U.S. EPA 2003), and The Consolidated Assessment and Listing
Methodology (CALM). Toward a Compendium of Best Practices (U.S. EPA 2002b), both of
which provide a framework for documenting the collection and use of water quality data for
CWA Section 305(b) reporting, determining attainment of WQS, determining stream impairment
for CWA Section 303(d) listing, and establishing anti-degradation policies.
This document attempts to follow the general framework provided by the CALM
document to organize the information for each of the Region 1 states in a format that is
conceptually easy for comparisons to be made among biomonitoring programs.
1.2.5 Biological Monitoring Programs
After beneficial uses are established, the criteria are set, and the anti-degradation policy is
in place, each state then implements a monitoring program. Bioassessment programs have been
employed by states to assess the water quality with established biocriteria for a range of
designated uses in freshwater systems. Bioassessment is used for a number of designated uses,
which may include drinking water, recreation, industry, wildlife, agriculture, and others, but it is
most commonly used to evaluate aquatic life use support. In 1991, U.S. EPA policy stated the
necessity of integrating biological surveys with toxicity and chemical-specific assessment
methods into monitoring programs to determine the attainment or non-attainment of aquatic life
use support (U.S. EPA 1991). As of 2001, 40 entities, including all of the Region 1 states, used
bioassessment to determine ALUS for 305(b) reporting (U.S. EPA 2002c).
Currently, the U.S. EPA CALM guidance suggests four categories of data that may be
collected and integrated to determine ALUS. These four categories are: biological, habitat,
lexicological, and physical/chemical data (U.S. EPA 2002b). Although all categories of data are
potentially useful depending on the rigor involved in the assessment method, only biological data
provide a direct measurement of the resident aquatic organisms that integrates the abiotic
conditions in the water body (U.S. EPA 2002b). The CALM document advises that states use
biological data "as a core indicator for aquatic life use determinations, as they are a unique water
body response measurement, providing information about a water body that no other
measurement can" (U.S. EPA 2002b). The document continues to stress that the state
"documentation of the adequate quality and rigor of the key elements of the state's
bioassessment program" be provided so that the biological data can accurately assess water
quality (U.S. EPA 2002b).
1.2.6 Bioindicator Organisms
Biological assessments of those organisms present in the aquatic system offer the most
direct way to measure the condition of the biological community as a function of environmental
stressors (Yoder and Rankin 1995). Community composition may be altered as a result of
stresses in the system and the condition of individual organisms can show pollution impacts that
may act as an early warning detection of degradation or provide a more reliable assessment of
changes in the biological community over time (U.S. EPA 2002d). There are several possible
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assemblages of organisms available for use in bioassessment. However, benthic
macroinvertebrates, periphyton, and fish are the biological indicators suggested for use by the
U.S. EPA in lotic environments (Barbour et al. 1999). All three assemblages are widely used in
bioassessment, but macroinvertebrates and fish are the most common indicator organisms, with
45 entities using two to four assemblages (U.S. EPA 2002c). Although standard methods for
sampling each of these assemblages are suggested by the U.S. EPA, many states alter the
methods to fit into the goals of their program, adjust for ecoregional constraints, and
accommodate limited budgets.
Benthic macroinvertebrates are the most commonly used assemblage. As of 2001, all 57
of the entities with a bioassessment program in place either currently used or were developing
macroinvertebrate indicators (U.S. EPA 2002c). Benthic macroinvertebrates are a diverse
assemblage, consist of species exhibiting a range of pollution tolerance levels, and are abundant
in most streams (Plafkin et al. 1989, Barbour et al. 1999). Furthermore, they often live the
majority of their lives in direct contact with both the water and sediments and their life cycles
may span multiple seasons, thereby showing cumulative changes. They also serve as an
important link in the food chain (Plafkin et al. 1989), maintaining the rest of the aquatic
community and managing algal systems. Benthic macroinvertebrates are easy and affordable to
collect, making them extremely attractive for biological monitoring.
The advantage of using periphyton as an indicator is that growth of this assemblage is
directly related to nutrient eutrophication and this assemblage may show adverse effects of
herbicides or other chemicals more quickly than other organisms (Barbour et al. 1999).
Periphyton assemblages exhibit stressor-related changes that alter species composition rapidly,
and can shift to noxious levels of overgrowth, thereby contributing to water quality degradation
(Stevenson et al. 1996, Stevenson and Bahls 1999). Similar to benthic macroinvertebrates,
periphyton assemblages contain species with a wide range of pollution tolerances. Furthermore,
they are easy to collect and identify by experienced taxonomists (Plafkin et al. 1989, Stevenson
and Bahls 1999). As of 2001, only 20 entities were using algae as an indicator, although an
additional 5 entities were developing algal indicators (U.S. EPA 2002c).
Fish are another indicator of watershed health with easily identifiable species of varying
trophic levels that respond differently to wide ranges of environmental stressors (Karr et al.
1986, Barbour et al. 1999). Fish are advantageous indicators because they live their entire lives
in water and their large geographical ranges can indicate the effects of stressors on a greater scale
than either periphyton or macroinvertebrates. Fish provide information regarding the physical,
chemical, biological and habitat condition of the watershed as a whole
(www.epa.gov/bioindicators/html/fish.html). As of 2001, 41 entities were using fish for
biological assessments (U.S. EPA 2002c).
1.2.7 305 (b) Report and 303 (d) List
Following data collection, processing, and analysis, each state is required to submit the
results in the form of a biennial 305(b) Report on the water quality conditions and provide a
303(d) List of Impaired Waters on April 1st of every even-numbered year (U.S. EPA 2003). The
305(b) report must contain all the information collected from streams and rivers located within
the state's boundaries. The Integrated 305(b) Report must contain the following key
components: "geographic referencing of all water resources; categorization of waters according
to WQS attainment status; identification, prioritization and scheduling of waters needing Total
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Maximum Daily Loads (TMDL); identification of waters where information is not sufficient to
determine a water's status; and a schedule of monitoring for the next reporting cycle" (U.S. EPA
2003). The EPA requires that all of the state's assessed waters be placed into one of five
categories that represent varying levels of water quality standards attainment. These five
categories as stated in U.S. EPA (2003) are as follows:
Category 1: All designated uses are met;
Category 2: Some of the designated uses are met but there is insufficient data to determine if
remaining designated uses are met;
Category 3: Insufficient data to determine whether any designated uses are met;
Category 4: Water is impaired or threatened but a TMDL is not needed;
Category 5: Water is impaired or threatened and a TMDL is needed.
Those impaired streams where one or more designated uses are not attained and are
consequently placed in Category 5 must be listed on the 303(d) list. Once placed on the 303(d)
list, a TMDL must be prioritized and established. Within the 303(d) list, Section 130.7(b)(4)
requires that each state also identify the pollutants that are known to be causing the impairment
(U.S. EPA 2003).
1.3 Literature Cited
Barbour, M, J. Gerritsen, B.D. Snyder, and J.B. Stribling. 1999. Rapid Bioassessment
Protocols for Use in Streams and Wadeable Rivers: Periphyton, Benthic
Macroinvertebrates and Fish, Second Edition. EPA 841-B-99-002. U.S. Environmental
Protection Agency, Office of Water, Washington, D.C.
http://www.epa.gov/owow/monitoring/rbp/wp61 pdf/rbp. pdf
(CT DEP) Connecticut Department of Environmental Protection. 2002a. Water Quality
Standards. Connecticut Department of Environmental Protection, Hartford, CT.
http://dep.state.ct.us/wtr/wq/wqs.pdf
Karr, J.R., K.D. Fausch, P.L. Angermeier, P.R. Yant, and IJ. Schlosser. 1986. Assessing
biological integrity in running waters: A method and its rationale. Special publication 5.
Illinois Natural History Survey.
(MDEP) Maine Department of Environmental Protection. 2003. Classification Attainment
Evaluation Using Biological Criteria for Rivers and Streams. Rules of Maine State
Government Agencies, 06 096 Chapter 579.
http://www.maine.gov/sos/cec/rcn/apa/06/096/096c579.doc
Maine Revised Statutes Annotate. Title 38, Chapter 3: Protection and Improvement of Waters.
Sections 464 and 465. http://janus.state.me.us/legis/statutes/38/title38ch3sec0.html
(MA DEP) Massachusetts Department of Environmental Protection. 2003. Massachusetts Year
2002 Integrated List of Waters: Part 1, Context and Rationale for Assessing and
Reporting the Quality of Massachusetts Surface Waters. CN: 125.1. Massachusetts
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Department of Environmental Protection, Bureau of Resource Protection, Division of
Watershed Management, http://www.mass.gov/dep/brp/wm/files/2002-il 1 .pdf
MA DEP. 2000. 314 CMR 4.00, Massachusetts Surface Water Quality Standards. Massachusetts
Department of Environmental Protection, Division of Water Pollution Control.
http://www.mass.gov/dep/bwp/iww/files/314cmr4.htm
(NH DES) New Hampshire Department of Environmental Services. 1999. State of New
Hampshire Surface Water Quality Regulations, Chapter 1700. New Hampshire
Department of Environmental Proteciton, Concord, NH.
http://www.des.state.nh.us/wmb/env-wsl700.pdf
Plafkin, J.L., M.T. Barbour, K.D. Porter, S.K. Gross, and R.M. Hughes. 1989. Rapid
Bioassessment Protocols for Use in Streams and Rivers: Benthic Macroinvertebrates and
Fish. EPA/444/4-89-001. U.S. Environmental Protection Agency, Washington DC.
(RI DEM) Rhode Island Department of Environmental Management. 2000. Water Quality
Regulations, Regulation EVM 112-88.97-1. Rhode Island Department of Environmental
Management, Office of Water Resources.
http://www.state.ri.us/dem/pubs/regs/REGS/WATER/h20qltv.pdf
Stevenson, R.J., and L.L. Bahls. 1999. Periphyton protocols. Pages 6-1 to 6-22 in M.T.
Barbour, J. Gerritsen, and B.D. Snyder, and J.B. Stribling (editors). Rapid Bioassessment
Protocols for Use in Wadeable Streams and Rivers: Periphyton, Benthic
Macroinvertebrates, and Fish, Second Edition. EPA 841-B-99-002. United States
Environmental Protection Agency, Washington, D.C.
Stevenson, R.J., M.L. Bothwell, and R.L. Lowe. 1996. Algal Ecology: Freshwater Benthic
Ecosystems. Academic Press, New York.
(U.S. EPA) U.S. Environmental Protection Agency. 1991. Policy on the Use of Biological
Assessments and Criteria in the Water Quality Program. U.S. Environmental Protection
Agency, Office of Water, Washington, D.C.
http://www.epa.gov/bioindicators/pdf/bioass_policy.pdf
U.S. EPA. 2002a. National Recommended Water Quality Criteria. EPA-822-R-02-047. U.S.
Environmental Protection Agency, Office of Water, Washington D.C.
http://www.epa.gov/waterscience/pc/revcom.pdf
U.S. EPA. 2002b. Consolidated Assessment and Listing Methodology, Toward a Compendium
of Best Practices, 1st Edition. U.S. Environmental Protection Agency, Office of Water,
Washington, D.C. http://www.epa.gov/owow/monitoring/calm.html
U.S. EPA. 2002c. Summary of Biological Assessment Programs and Biocriteria Development
for States, Tribes, Territories, and Interstate Commissions: Streams and Wadeable Rivers.
EPA-822-R-02-048. U.S. Environmental Protection Agency.
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U.S. EPA. 2002d. Biological Assessments and Criteria: Crucial Components of Water Quality
Programs. EPA 822-F-02-006. U.S. Environmental Protection Agency, Office of Water,
Washington D.C. http://www.epa.gov/ost/biocriteria/technical/brochure.pdf
U.S. EPA. 2003. Guidance for 2004 Assessment, Listing and Reporting Requirements Pursuant
to Sections 303(d) and 305(b) of the Clean Water Act; TMDL-01-03. U.S.
Environmental Protection Agency, Office of Water, Washington, D.C.
http://www.epa.gov/owow/tmdl/tmdl0103/2004rpt_guidance.pdf
State of Vermont. 2000. Vermont Water Quality Standards. State of Vermont, Water
Resources Board, Montpelier, VT. http://www.state.vt.us/wtrboard/julv2000wqs.htm
Yoder, C.O., and E.T. Rankin. 1995. Biological criteria program development and
implementation in Ohio. Pages 109-144 in W.S. Davis and T.P Simon (editors).
Biological Assessment and Criteria: Tools for Water Resource Planning and Decision
Making. Lewis Publishers, Boca Raton, FL.
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2 CONNECTICUT
This document was prepared using documents written by the State of
Connecticut and via Personal Communication with the State
Supervising Environmental Analyst. Any questions concerning
bioassessment methods should be directed to:
Ernest Pizzuto, Jr., Supervising Environmental Analyst
Connecticut Department of Environmental Protection (CT DEP)
79 Elm Street
Hartford, CT 06106-5127
Phone: (860) 424-3715; Fax ((860) 424-4055
Email: Ernest.Pizzuto@po.state.ct.us
2.1 Introduction
The CT DEP Bureau of Water Management has used the benthic macroinvertebrate
assemblage to assess the biological integrity of surface waters since the mid-1970's and began
using fish assemblage data in 1999 in cooperation with the CT DEP Inland Fisheries Division.
The benthic macroinvertebrate assemblage is assessed based on the Rapid Bioassessment
Protocol (RBP) III Single Habitat method (Plafkin et al. 1989, Barbour et al. 1999), and an index
modified from Plafkin et al. (1989) is used to determine the level of ALUS (i.e., Full Support,
Threatened, Partial Support, Not Supporting). Connecticut WQS state that "the benthic
invertebrate criteria may be utilized where appropriate for assessment of the biological integrity
of surface waters. The criteria apply to the fauna of erosional or riffle habitats in lotic waters
which are not subject to tidal influences" (CT DEP 2002a). In addition to the biological
component, habitat, aquatic toxicity, sediment, and ambient chemical and physical data collected
by the Connecticut Ambient Biological Monitoring Program are used to determine compliance
with State WQS (Table 2-1) and are ultimately used to report on the ALUS under section 305(b)
and 303(d) of the CWA. Furthermore, the ambient monitoring program seeks to evaluate
pollution control program effectiveness, collect data for baseline characterization and
identification of reference conditions, assess water quality trends, evaluate ecological damage
due to emergency pollution events, identify existing and emerging pollution problems, and
investigate nuisance complaints (CT DEP 1999).
2.2 Key Elements of the Biological Assessment Approach
2.2.1 Index Period and/or Temporal Conditions
Biological monitoring by CT DEP utilizes benthic macroinvertebrates as the primary aquatic
assemblage for ALUS assessment purposes. Fish assemblage data have been incorporated on a
limited basis since 1999. Based on differences in the biology of these indicator assemblages and
logistical considerations, different index periods have been selected for their collection. Benthic
macroinvertebrate data are collected in the late autumn (October 1-December 1). This time
frame provides for the collection of individuals that are large enough to identify. It also
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Table 2-1. Connecticut water quality standard classes.
Class
Management
Biological Standard
A
Designated as a
potential drinking
water supply, fish and
wildlife habitat,
recreation, industrial
supply and other
legitimate uses,
including navigation.
A wide variety of macroinvertebrate taxa should
normally be present and all functional feeding groups
should normally be well-represented. Presence and
productivity of aquatic species are not limited except
by natural conditions, permitted flow regulation or
irreversible cultural impacts. Water quality shall
be sufficient to sustain a diverse macroinvertebrate
community of indigenous species. Taxa within the
orders Plecoptera (stoneflies), Ephemeroptera
(mayflies), Coleoptera (beetles), and Trichoptera
(caddisflies) should be well-represented.
B
Designated as a use for
habitat for fish and
other aquatic life and
wildlife, recreation,
navigation, and
agricultural and
industrial water supply.
Water quality shall be sufficient to sustain a diverse
macroinvertebrate community of indigenous species.
All functional feeding groups and a wide variety of
macroinvertebrate taxa shall be present; however, one
or more may be disproportionate in abundance.
Waters which currently support a high quality aquatic
community shall be maintained at that high
quality. Presence and productivity of taxa within the
orders Plecoptera, Ephemeroptera; and pollution
intolerant Coleoptera and Trichoptera may be limited
due to cultural activities. Macroinvertebrate
communities in waters impaired by cultural activities
shall be restored to the extent practical through
implementation of the department's procedures for
control of pollutant discharges to surface waters and
through Best Management Practices (BMPs) for non-
point sources of pollution.
C
Suitable for certain fish
and wildlife habitat,
recreational activities,
industrial use, and
other legitimate uses,
including navigation.
Not defined in WQS
D
May be suitable for
bathing or other
recreational
purposes, certain fish
and wildlife habitat,
industrial uses and
navigation.
Not defined in WQS
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provides the worst-case scenario for impairment of waste-receiving waters, and allows for
conclusions of this impairment to be drawn based on the macroinvertebrate assemblage
assessment. Fish monitoring is conducted during the summer low flow period. This is a period
of high stress for fish assemblages in Connecticut streams, and low stream flow facilitates
sample collection. In 2002-2003, CT DEP was funded by U.S. EPA to collect periphyton data
for a pilot project. It was the intention of CT DEP to use the project for incorporating the
methods developed into the ambient biological monitoring program in the future. For the pilot
project, periphyton was collected in July and August, the period of stable flow and high
periphyton growth rates (due to maximum ambient temperature from increased available sunlight
and day length).
2.2.2 Monitoring Program Survey Approach
Water quality monitoring in Connecticut has historically employed a focused approach
targeting major rivers and waste receiving waters. Consequently, many smaller streams
remained unassessed. In an effort to prioritize surface water monitoring activities and increase
monitoring coverage, a five-year rotating basin monitoring strategy was developed and
implemented in 1997. One major drainage basin was targeted each year during the five-year
cycle that ended in 2001. Within each basin, approximately fifty sites were sampled annually.
Criteria used to select sites for sampling were sub-basin size, location of wastewater discharges,
land use, and resource value. A subset of approximately 24 targeted sites was chosen each year
to assess the fish assemblage. Additionally, an increased effort was made to incorporate data
from volunteers, academics and municipalities. To work toward the goal of a comprehensive
assessment, the CT DEP accepted the opportunity to participate with U.S. EPA in a two-year
monitoring project following completion of the five-year rotating basin strategy in 2001. This
project was conducted during 2002 and 2003 and assessed wadeable streams based on a
statewide probabilistic design. Sample coverage included monitoring of macroinvertebrates at
60 sites, fish at 24 sites, and periphyton at 30 sites. Water samples were collected quarterly for
chemical analyses at the 60 sites sampled for benthos. The CT DEP is currently developing a
Comprehensive Monitoring and Assessment Strategy to meet CWA Section 106 requirements.
This strategy will be completed by October 2004 and will cover a ten-year period. It will include
elements of the previous rotating basin strategy as well as a probabilistic component.
2.2.3 Indicator Assemblages
Currently, CT DEP primarily uses benthic macroinvertebrates as the indicator
assemblage for biological monitoring. The CT DEP incorporates fish assemblage data using best
professional judgment to make decisions about class attainment and is currently developing a
Fish Index of Biotic Integrity (IBI) based on the State of Vermont model. A pilot project for
periphyton sampling was conducted in 2002-2003, and the data were used only to supplement
other biological data. Methods for the use of periphyton in the monitoring program are under
development.
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2.2.4
Reference Condition
The CT DEP has selected reference sites to compare to test sites within each of
Connecticut's major basins (Figure 2-1). Those sites designated as reference have been defined
as those that are least disturbed and minimally impacted by human influences. Furthermore,
reference sites are selected for comparison against test sites so that the streams are within one
stream order (±1) and drainage areas are within one order of magnitude of one another.
Reference sites are also used for comparison if the stream gradient is similar to that of the test
site. The natural features of a reference site should include wadeable streams with optimal
habitat including a hard bottom and erosional substrate (i.e., riffle habitat with cobble or gravel
substrate).
Lower ah amev SoutheastiKoastal, Pa'wcatuck
Southwest Coastal and Soj/WCentral Coastal
Figure 2-1. Major Connecticut basins sampled for the biological monitoring program
using the rotating basin strategy.
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2.3 Field and Laboratory Protocols
2.3.1 Macroinvertebrate Protocols
2.3.1.1 Field Methods
The CT DEP uses a modified RBP Single Habitat Approach to collect macroinvertebrate
samples (Plafkin et al. 1989, Barbour et al. 1999). After twelve riffle sampling points ("stops")
are chosen in a sampling reach, a rectangular kick net (9 in x 18 in) with 800-900 jim mesh,
modified from the RBP recommended 500 jim, is placed at the bottom of each riffle with the
opening facing upstream. An approximately 2-m2 area of substrate upstream of the net is
disturbed at each riffle using a kicking and stomping motion and the loosened
macroinvertebrates are then trapped in the net. The sample is removed, the net is repositioned at
different riffles or within the same riffle, and the process is repeated until all twelve samples are
collected. The twelve samples are then composited into one jar, labeled and preserved with 70%
ethanol.
2.3.1.2 Laboratory Methods
Benthic macroinvertebrate samples are rinsed through a #30 sieve (600 jim) to remove
preservative and any large debris. The entire sample is then spread over a gridded tray
containing 56 squares. Enough water is added to moisten organisms and spread the sample
evenly. Random squares are selected to sort completely until 200 organisms (± 10%) are
counted. After the sample is subsampled in this way, any large, rare or representative taxa are
removed from the remaining debris. The sample is preserved with 70% ethanol. The sample is
then identified by a qualified taxonomist using dissecting microscopes (10x-64x) to the lowest
possible taxonomic classification using various keys and Peckarsky et al. (1990). Chironomids
are placed in 15% KOH overnight, mounted on slides using glycerine, and then identified using a
compound microscope. The CT DEP maintains a reference collection of taxa and any taxa
unable to be identified by CT DEP taxonomists are verified by regional taxonomists.
2.3.2 Periphyton Protocols (CT DEP 2003)
2.3.2.1 Field Methods
Two methods were used to assess benthic algae during 2002-2003. Samples were
collected using a modified RBP Single Habitat sampling protocol (Barbour et al. 1999) for
natural substrates to assess algal biomass and taxonomic composition. In addition, the field-
based Rapid Periphyton Survey (RPS) (Barbour et al. 1999) was conducted.
2.3.2.1.1 Quantitative Periphyton Sampling
At each site, a 150-m reach is selected for assessment and fifteen pieces of substrate
(ideally rocks) with sizes of 6.4-25.6 cm diameter are collected from throughout the reach. If
rocks are not present, then logs and large sticks are used. The fifteen pieces of substrate are
carried to the bank, and attached algae are scraped from a 1-in diameter area using a flat spatula
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and a toothbrush. A rinse bottle filled with deionized water is used to wash the algal material
into a Nalgene® collection bottle. The algae from all 15 pieces of substrate are composited into a
single sample bottle. The bottle is placed on ice and returned to the lab where 5 ml of sample is
removed for chlorophyll a analysis. The remaining sample is then preserved with 2% formalin
and analyzed to determine the identification and biomass of the sampled periphyton. A duplicate
sample is collected for quality control at 10% of sites.
2.3.2.1.2 Rapid Periphyton Survey (Barbour et al. 1999, CT DEP 2003)
At each site, the width of the stream is estimated to establish the number of transects that
will be laid out (5-25). Then, transects are divided into observation points ("stops") to be
sampled, so that 2-10 stops are sampled at each transect. A viewing bucket with a 50 dot grid is
immersed in the stream and at each stop the observer visually estimates and records the number
of grid points covered by moss, macroalgae and microalgae. The observer also records the
distance from the left bank, depth, dominant substrate, canopy cover, current velocity, size of the
average rock/substrate within 1 ft of the observation point, and the presence or absence of
vascular plants. All data are recorded on the CT RPS Data Sheet (CT DEP 2003).
2.3.2.2 Laboratory Methods (CT DEP 2003)
2.3.2.2.1 Chlorophyll a (APHA 1999)
After arriving at the lab, the 5 ml subsample removed from the composite periphyton
sample is filtered through a 47 mm GF/F filter with a nominal pore size of 0.7 mm. The filter is
then stored in an aluminum foil packet and frozen at -15°C until transfer to a laboratory for
processing (Environmental Research Institute at the University of Connecticut) within three
weeks of filtration. The pigment is extracted from the filter using an aqueous acetone MGCOs
solution. Chlorophyll a concentration is determined using the U.S. EPA Method AERP-12
fluorometric method. A Turner 450 Flourometer with a 1-cm light path length is used. The
fluorometer is calibrated by using a chlorophyll a standard of known value, which is also run by
spectrophotometric method. The fluorescence of the extract is determined and the chlorophyll a
concentration is calculated using the following equation:
Chla mg/m2 = (Ca)(Extract volume in L)/(Substrate area in m2 represented by filter)
where:
Ca = Concentration of Chla in mg I L =
T^I 7- /• TI • f ^ ugChla mgChla
Fluorometer reading x (instrument calibration jactor ) = —2 =—
Extract Volume in L = always equals 0.02 L
07 77 r-i \5ml filteredvolume•}. , 7 ~ ™-™~,- 2-.
Substrate area represented by jitter = \ \(total substrate area 0.0076035m )
^ total volume in ml j
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2.3.2.2.2 Algal Identification and Density (contracted to Dr. R. Jan Stevenson
at Michigan State University)
A Palmer Counting Cell is used to count at least 300 cells and identify soft algae to
species or to subspecies/variety (rarely to genus), and count live diatoms using the
Environmental Monitoring and Assessment Program (EMAP) protocol (Lazorchek et al. 2000,
Charles et al. 2002). Next, diatom valves are acid cleaned and mounted in NAPHRAX. Then,
600 diatom valves are identified to species or lower level in a second count. Algal densities per
unit area of substratum and relative abundances of algal taxa are calculated as advised in the
RBPs for algae (Barbour et al. 1999).
2.3.2.2.3 Biomass and Biovolume Determination
Biomass, or Ash Free Dry Mass (AFDM), is determined using Standard Method 10300C
(APHA 1999). Biovolumes of algae are determined by measuring at least 15 cells of each taxon
that occurs with a relative abundance greater than 5% in any sample. For rarer taxa, fewer cells
are measured to determine their biovolume. For taxa for which all measures cannot be made or
taxa with less than a 1% average relative abundance among all samples, literature values or
database values may be used to determine species biovolumes. For each sample, the relative
biovolumes of taxonomic and functional groups (as defined by algal class and growth form:
centric diatom, pennate diatom, filamentous cyanobacteria, filamentous green algae, etc.) are
calculated.
2.3.3 Fish Protocol (Plafkin et al. 1989, Hagstrom et al. 1995, CT DEP 2002b)
Fish assemblage sampling is conducted in cooperation with the CT DEP Inland Fisheries
Division. At each site, a 150-m reach is selected for assessing the fish assemblage. The upper
and lower boundaries of the reach are determined by natural barriers to fish movement. The CT
DEP varies the electrofishing equipment depending on the width of the stream to be sampled. A
single backpack is used in the smallest streams, two backpacks are used in medium-sized
streams, one generator towed in a canoe is used in large streams, and two or three generators are
towed in multiple canoes for the very largest streams. In all cases, the elecrofishing crew
(minimally consisting of three people), begin at the downstream barrier and move upstream
collecting all species affected after one pass. All fish that are greater than 3 cm in length are
measured, identified, and the condition and any anomalies are recorded in the field before they
are returned to the stream. Any fish that cannot be identified in the field are sent to the
laboratory (alive or preserved in ethanol) to be identified by a senior field biologist using
Whitworth(1996).
2.4 Data Management/Quality
A Microsoft Access database is used to track all sample collection, analyses, and
resultant metadata. All data are linked to sampling trips by unique sample identifiers and each
location is stored as a unique point and gee-referenced. All metadata are entered in the correct
sequence to keep sample results linked with sample metadata; electronic transfer of results is
used whenever possible to reduce transcription error. The database is linked to Arc View GIS
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software to enable the mapping and graphic analysis of data. Ultimately, all data are stored in
the U.S. EPA Storage and Retrieval database (STORET), a repository for water quality,
biological, and physical data.
2.5 Analysis of Biological Data
2.5.1 Macroinvertebrate Data
For the macroinvertebrate assemblage, RBP III thresholds as described in Table 2.2 are
applied to seven metrics (Plafkin et al. 1989) to determine metric scores. The sum of metric
scores is then represented as a percent of the reference total score, and the test stream is placed
into one of four impact categories: Not Impaired, Slightly Impaired, Moderately Impaired, or
Severely Impaired (Table 2-2). The CT DEP recognizes any test stream score of less than 54%
as not fully supporting ALU, with a gray area lying between 50-54%. The streams in the gray
area are placed in categories on a case-by-case basis weighted on the evidence and available
data.
Table 2-2. Metrics and scoring ranges used in RBP III determinations of the level of
biological impact based on benthic macroinvertebrates (based on Plafkin et al. (1989)).
Metric
Taxa Richness (a)
EPT Index (a)
EPT/ Chironomidae
(abundance ratio) (a)
HBI (modified) (b)
Scraper/Filtering
Collector Ratio (a)
% Contribution of
Dominant Taxon (o)
Method
The total number of distinct taxa in
a sample
The number of taxa within the
orders of Ephemeroptera,
Plecoptera, and Trichoptera
(Abundance of EPT organisms)/
(Abundance of EPT +
Chironomidae)
(Number of individuals in each
taxon multiplied by its assigned
tolerance value)/(Total number of
organisms in sample)
(Number of scrapers)/(Number of
filtering organisms)
(Number of individuals in most
common taxon)/(Total number of
organisms) x 100
Scoring Ranges
6
>80%
>90%
>75%
>85%
>50%
<20%
4
60-80%
80-90%
50-75%
70-85%
35-50%
20-29%
2
40-59%
70-79%
25-49%
50-69%
20-34%
30-40%
0
<40%
<70%
<25%
<50%
<20%
>40%
2-8
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Metric
Method
Scoring Ranges
Community Loss
Index(d)
A measure of the dissimilarity
between a test site and a reference
site (Plafkin et al. 1989). Metric
values increase as biological
impairment increases. Values have
no limits.
CLI = a-c/b
where: a = number of genera in
reference sample, b = number of
genera in test sample, c = number of
genera common to both samples
<0.5
0.5-1.5
1.6-4.0
>4.0
% Total Observed Score compared to Reference Condition
> 83% Not Impaired
54-79% Slightly Impaired
21-50% Moderately Impaired
<17% Severely Impaired
(a) Value is converted to ratio of test to reference site * 100
(b) Value is converted to ratio of reference to test site * 100
(c) Actual percent contribution used in scoring, not ratio to reference
(d) Uses range of values actually obtained
2.5.2 Periphyton Data
The CT DEP is under the process of developing algae as an indicator using probabilistic
sampling in their 2002-2003 pilot study. The data collected were used to supplement other
biological data collected while the methods are under development. Literature values were used
to calculate metrics during the first year of study. However, autoecological information for
species within Connecticut populations can be generated with data from the pilot study and then
tested using data from the second year of the study. Metrics calculated are derived from RBP
periphyton protocols (Barbour et al. 1999). Data that are generated from the RPS and data from
Chlorophyll a, biomass, and species composition and abundance will be evaluated to determine
the components that will be used in the routine ambient monitoring program.
2.5.3
Fish Data
Although CT DEP does not currently have a fish index, they are developing a Fish Index of
Biotic Integrity (IBI) based on the State of Vermont model. Until the IBI is developed the CT
DEP incorporates the results of the fish assemblage data using best professional judgment to
make decisions about class attainment. The data collected from fish assessments are species
composition, trophic structure and age class distribution and these measurements taken from
sampled streams are compared to those measurements in unimpaired and impaired streams to
make inferences about the condition of the fish assemblage.
2-9
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2.5.4 Summary: Determining ALU Support
Connecticut narrative criteria in WQS, the macroinvertebrate quantitative index, fish data
(while metrics are under development), and any other supplemental physical, chemical, and other
biological data are used to make the ALU assessment (listed in Table 2-3). These assignments
are then outlined in the 305(b) report and any streams not in attainment are placed on the
Connecticut 303(d) list. Table 2-3 outlines the guidelines CT DEP biologists use in conjunction
with BPJ to determine ALUS at sites.
Table 2-3. Aquatic life use support categories and the criteria used for making decisions
(taken from Table 2 in CT DEP 2002a).
ALUS
Criteria/Indicators
Fully Supporting
Benthic assemblage: bioassessment indicates assemblage is non-
impaired or slightly impaired (Plafkin et al. 1989), and meets
narrative criteria in CT WQS; RBP III Community Score > 54%
of reference condition.
Fish assemblage: species composition, trophic structure, and age
class distribution as expected for a non-impacted stream of
similar size.
Conventional physical/chemical criteria not exceeded.
Measured toxicants do not exceed chronic toxicity criteria.
No record of catastrophic events (e.g., chemical spills, fish kills)
No evidence of flow diversion
Threatened
Benthic assemblage: non-impaired or slightly impaired, but still
meets narrative criteria in CT WQS; RBP III Community Score
>54% reference condition.
Fish assemblage as above, but documented trend is downward or
conditions exist that may impact the assemblage in the future.
Slight exceedences of either conventional or toxicant criteria in <
10% of samples; exceedences difficult to discern from expected
analytical variability or error.
Discharge effluent constitutes >20% of stream flow.
Land use conditions exist that may cause impairment.
Flow reductions due to diversions have been observed.
Partially
Supporting
Benthic assemblage: bioassessment indicates assemblage is
moderately impaired; RBP III Community Score 21-50% of
reference condition.
Fish assemblage: species composition, trophic structure and age
class distribution significantly less than expected for non-
impacted stream of similar size: diversity and abundance of
intolerant species reduced; top carnivores rare; trophic structure
skewed toward omnivory.
Either fish or benthic assemblage meets above conditions, but the
other assemblage is fully supporting.
Conventional physical/chemical criteria exceeded in > 10% but <
2-10
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ALUS
Not Supporting
Not Attainable
Criteria/Indicators
25% of samples.
• Measured toxicants exceed chronic criteria < 10% of samples.
• Flow is reduced significantly during drought conditions.
• Benthic assemblage: bioassessment indicates assemblage is
severely impaired: RBP III Community Score < 17% of
reference condition.
• Fish assemblage: species composition, age class distribution and
trophic structure greatly impaired in comparison to a non-
impacted or minimally impacted stream of similar size;
assemblage dominated by highly tolerant species, omnivores and
habitat generalists; in extreme cases, few species present and/or
diseased fish common.
• Conventional physical/chemical criteria exceeded in > 25% of
samples
• Measured toxicants exceed chronic criteria >10% of samples
• Stream known to dry completely for significant periods.
• Documented catastrophic event (e.g., chemical spill, fish kill)
Stream completely enclosed in conduit or cleared concrete trough.
Stream is dewatered most of the time due to an upstream
impoundment or diversion.
2.6
Literature Cited
(APHA) American Public Health Association. 1999. Standard Methods for the Examination of
Water and Wastewater, 20th edition. American Public Health Association, Washington,
D.C.
Barbour, M, J. Gerritsen, B.D. Snyder, and J.B. Stribling. 1999. Rapid Bioassessment
Protocols for Use in Streams and Wadeable Rivers: Periphyton, Benthic
Macroinvertebrates and Fish, Second Edition. EPA 841-B-99-002. U.S. Environmental
Protection Agency, Office of Water, Washington, D.C.
Charles, D.F., C. Knowles and R.S. Davis (editors). 2002. Protocols for the Analysis of Algal
Samples Collected as Part of the U.S. Geological Survey National Water-Quality
Assessment Program. Report No. 02-06. The Academy of Natural Sciences,
Philadelphia, PA.
(CT DEP) Connecticut Department of Environmental Protection. 1999. Ambient Monitoring
Strategy for Rivers and Streams: Rotating Basin Approach. CT Department of
Environmental Protection, Bureau of Water Management, Planning and Standards
Division, Hartford, CT. http://dep.state.ct.us/wtr/wq/rotbasinplan.pdf
CT DEP. 2002a. Water Quality Standards. Connecticut Department of Environmental
Protection, Bureau of Water Management, Planning and Standards Division, Hartford,
CT. http://dep.state.ct.us/wtr/wq/wqs.pdf
2-11
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CT DEP. 2002b. Quality Assurance Project Plan: Ambient Biological Monitoring - Fish
Community Structure. Prepared by Michael Beauchene, CT Department of
Environmental Protection, Bureau of Water Management, Planning and Standards
Division, Hartford, CT.
CT DEP. 2003. Quality Assurance Project Plan: Ambient Biological Monitoring: Periphyton
and Chlorophyll Concentrations. Prepared by Lisa Wahle, CT Department of
Environmental Protection, Bureau of Water Management, Planning and Standards
Division, Hartford, CT.
Lazorchak, J.M., B.H. Hill, O.K. Averill, D.V. Peck, and DJ. Klemm (editors). 2000.
Environmental Monitoring and Assessment Program-Surf ace Waters: Field Operations
and Methods for Measuring the Ecological Condition of Non-wadeable Rivers and
Streams. EPA 620/R-00/007. U.S. Environmental Protection Agency, Cincinnati, OH.
Hagstrom, N.T., M. Humphreys, W.A. Hyatt, and W.B. Gerrish. 1995. A Survey of Connecticut
Streams and River: Statewide Summary. F-66-R: Final Report. Connecticut Department
of Environmental Protection, Federal Aid in Sport Fish Restoration.
Peckarsky, B.L., P.R. Fraissinet, M.A. Penton, and DJ. Conklin, Jr. 1990. Freshwater
Macroinvertebrates of Northeastern North America. Cornell University Press, Ithaca,
NY.
Plafkin, J.L., M.T. Barbour, K.D. Porter, S.K. Gross, and R.M. Hughes. 1989. Rapid
Bioassessment Protocols for Use in Streams and Rivers: Benthic Macroinvertebrates and
Fish. EPA/444/4-89-001. U.S. Environmental Protection Agency, Washington, D.C.
Whitworth, W. 1996. Freshwater Fishes of Connecticut. CT DEP Bulletin number 114, 2nd
edition. State Geologic and Natural History Survey, Connecticut Department of
Environmental Protection, Hartford, CT.
2.7 Resources
CT DEP Web Page http://dep.state.ct.us/index.htm.
U.S. EPA. Biological Indicators of Watershed Health, Connecticut Webpage:
http://www.epa.gov/bioindicators/html/state/ct-bio.html
U.S. EPA. 2002. Summary of Biological Assessment Programs and Biocriteria Development
for States, Tribes, Territories, and Interstate Commissions: Streams and Wadeable Rivers.
EPA-822-R-02-048. U.S. Environmental Protection Agency.
2-12
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MAINE
This document was prepared using documents written by Maine
Department of Environmental Protection personnel. Any questions
concerning bioassessment methods should be directed to:
Susan P. Davies, Program Manager, Biologist III
Maine Department of Environmental Protection
SHS17
Augusta, ME 04333
Phone: (207) 287-7778; Fax (207) 287-7191
Email: susan.p.davies@maine.gov
3.1 Introduction
Maine Department of Environmental Protection (MDEP) has developed a biological
monitoring and biocriteria program to assess water quality and ensure the adherence to ALU
designations defined in Maine's WQS. In 1983, MDEP began its standardized benthic
macroinvertebrate sampling program. This program began building a database to aid in the
development of numeric biocriteria using a discriminant model approach. The numeric
biocriteria developed were refined on a regional scale to increase the accuracy of measuring the
adherence of streams to aquatic life use standards. The biocriteria program was written into law
in April 1986 with the passing of M.R.S.A. 38 Public Law Chapter 698: The Classification
System for Maine Waters (State of Maine 1985). This law required the State to "restore and
maintain the chemical and biological integrity" of Maine waters. This law also established a
classification system, and narrative biological and habitat criteria were described for each of
these classes. Furthermore, the statute details specific numerical standards to which each class
must adhere for dissolved oxygen and bacterial concentrations (Table 3-1). The water quality
classes are AA and A, B, and C (Table 3-1). Water quality below Class C is given Non-
Attainment status.
Once water bodies were assigned a discrete water quality classification (i.e., A, B, C,
NA) and narrative aquatic life standards were established, MDEP began testing whether
empirical and ecological data collected from Maine's streams would show the gradients of
environmental quality reflected in the narrative standards (Figure 3-1). They were able to
conclude that the four established categories of biological condition did fit well with the State's
four-tiered standards for dissolved oxygen, bacteria and habitat (Davies et al. Draft). Multiple
exploratory multivariate analyses were performed, including k-means clustering,
multidimensional scaling, principal coordinate analysis, principal components analysis, multiple
regression analysis, two-way indicator species analysis, log-linear modeling, logistic regression,
detrended correspondence analysis, and variance component analysis, but MDEP ultimately
chose discriminant analysis to determine the probability of a stream's membership to an
established class (Davies et al. Draft).
3-1
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Table 3-1 Water quality classification system for rivers and streams in Maine. (M.R.S.A.
Title 38 Article 4-A § 464-465).
£
U
AA
A
B
C
Management
Highest quality water for
recreation and ecological
interests. Minimal human
influence. No discharges or
impoundments permitted.
High quality water with
limited human interference.
Discharges restricted to non-
contact process water or
highly treated wastewater
equal to or better than the
receiving water.
Impoundments allowed.
Good quality water.
Discharge of well-treated
effluent with ample dilution
permitted. Impoundments
allowed.
Acceptable water quality.
Maintains the interim goals
of the Federal Water Quality
Act (Fishable/swimmable).
Discharge of well-treated
effluent permitted.
Impoundments allowed.
Narrative Habitat and
Aquatic Life Standards
Habitat natural and free
flowing. Aquatic life as
naturally occurs.
Habitat natural. Aquatic life
as naturally occurs.
Habitat unimpaired.
Ambient water quality
sufficient to support life
stages of all indigenous
aquatic species. Only non-
detrimental changes in
community composition
allowed.
Habitat for fish and other
aquatic life. Ambient water
quality sufficient to support
life stages of all indigenous
aquatic species. Change in
community composition may
occur but structure and
function of the community
must be maintained.
Dissolved
Oxygen
Numeric
Criteria
As
naturally
occurs
7ppm;
75%
saturation
7ppm;
75%
saturation
5ppm;
60%
saturation
Bacteria
(E. coli)
Numeric
Criteria
As
naturally
occurs
As
naturally
occurs
64/100 ml
(geometric
mean) or
427/100
ml
(instantane
ous level)
142/100
ml
(geometric
mean) or
949/100
ml
(instantane
ous level)
* Classes AA and A are indistinguishable in the discriminant model because narrative criteria
are both described "as naturally occurs ".
5-2
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natural
Class ±4/A: Habitat Nature
Biological
Condition
degraded
Aquatic life as naturally occurs
Class B: Habitat unimpaired. Ambient
water quality sufficient to support life
stages of indigenous species. No
detrimental change allowed.
Class C: Ambient WO sufficient
to support life stages of all
indigenous fish species &
maintain structure & function.
Xot meeting CWA 101 a uses for protection &
propagation of aquatic life
High
Low Human Disturbance
Figure 3-1. Maine's narrative aquatic life standards with the human disturbance and
biological condition gradients (Taken from Courtemanch 2003).
The MDEP must report the specific class attainment of each stream under Section 305(b)
of the CWA. Those sites that are found in non-attainment must be listed on the state's 303(d)
list. For those streams placed on the 303(d) list, the MDEP is then expected to develop and
implement a total maximum daily load (TMDL) for each stressor that is preventing the stream
from reaching attainment status.
3.2
Key Elements of the Biological Assessment Approach
3.2.1 Index Period and/or Temporal Conditions
It is important to select a sampling season that is indicative of the conditions needed to
collect the most suitable data to answer the objectives for the intended study. Thus, the MDEP
has chosen the index sampling period to be July 1 - September 30th, the low flow period for
streams in Maine. All baseline data from streams were collected during this period because late
summer represents the time of the year when organisms may be exposed to maximum stressful
conditions (Davies and Tsomides 2002). During this period, water levels tend to drop, which
may increase the concentration of soluble contaminants or nutrients in the stream, and water
temperatures tend to be at a maximum level.
3-3
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3.2.2 Monitoring Program Survey Approach/Natural Classification of Water
Bodies
Maine has divided the state into 5 main watersheds that are sampled every five years on a
rotational basis. These basins are the Androscoggin River; Kennebec and Mid-Coast Basin;
Penobscot, St. Croix, and North Coastal Rivers; Piscataqua, Saco, and Southern Coast; and the
St. John and Presumpscot Basins (Figure 3-2). Although the water bodies have been divided into
five main watersheds for monitoring purposes, exploratory data analysis concluded that it was
not necessary to stratify the modeling approach climatically or geographically or to create
separate southern Maine and northern Maine models (Davies et al. Draft).
Approximately 50-60 sites are sampled each year within a single basin. Sites are chosen
based on a "targeted approach" that incorporates a variety of factors that document the
degradation or improvement of each stream. These factors include: 1) a prior knowledge of
existing activities that may degrade the water body and impact the biological community; 2) a
continued effort to monitor the effect a potential threat may have on a water body; 3) the
requirement (or scientific endeavor) to monitor remediation activities or water quality
management changes; and 4) to increase documentation of natural variability by including
previously unmonitored sites (MDEP 2002). Furthermore, the rotation schedule provides
assessment information for scheduled wastewater renewals.
3.2.3 Indicator Assemblage
Benthic macroinvertebrates were chosen as the biological endpoint because "they have
limited mobility; as a group, they include species representing a wide range of pollution
tolerances, including those found in extremely polluted sites; they are diverse and have relatively
long, complex life cycles; they are a food source for fish; methods of sample collection and
analysis are well-established for this assemblage; and they are a cost-effective group to sample"
(MDEP 1995, Davies et al. Draft).
3.2.4 Reference Condition (Establishing a priori Groups)
The MDEP used linear discriminant analysis for model construction, which requires that
a priori groups be established. For this reason, reference condition is more appropriately
addressed as a discussion of a priori groups. The MDEP used best professional judgment to set
these a priori groups (please see Section 3.5 Analysis of Biological Data and Appendix A for an
in-depth description of a priori group assignment).
Furthermore, MDEP does evaluate upstream and downstream of disturbed sites in order
to collect information about the biological condition before and after a disturbance. This
information is evaluated and used when determining the use attainment of a site (Davies et al.
Draft).
5-4
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Penobscol River
St. Croix Rive
North Coaslal Ri
Kennebec River
Anoroscoggin
Rive
Saco River
Pi scat a qua River
Figure 3-2 Map of basins sampled by MDEP (2002).
3.3 Field and Laboratory Protocols
3.3.1 Macroinvertebrate Protocols (from Davies et al. 1999, Davies and Tsomides
2002)
3.3.1.1 Field Methods
The MDEP follows a highly standardized and quantitative "Classification Attainment
Evaluation" protocol to collect data (see Davies et al. 1999, Davies and Tsomides 2002).
Depending on water depth encountered, the MDEP uses rock baskets, riffle bags, or rock-filled
cones to collect macroinvertebrates. Rock-filled mesh bags are used in small streams with
depths of at least 5 cm, and rock-filled cones are used in non-wadeable rivers that must be
accessed by boats for placement and retrieval. Rock baskets are used to sample wadeable
streams deep enough to allow the baskets to be fully submerged. Rock baskets consist of a
cylindrical wire barbecue basket filled with substrate. Each basket is constructed according to
U.S. EPA methods and has at least 1.5 cm spaces between wires, with a hinged opening and a
secure closure (Klemm et al. 1990). The substrate material is a clean cobble, relatively uniform
in diameter ranging from 3.8-7.6 cm. Each basket contains approximately 7.25 ± 0.5 kg of
3-5
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substrate. Rock baskets are placed in the riffle/run portion of a stream reach at a depth to ensure
that they will remain fully submerged. The apparatus is positioned in a portion of the stream that
reflects the overall characteristic of the stream sampled, located in the middle 50% of bank-to-
bank width or in an area that depicts the overall flow of the channel. At each site, a minimum of
three baskets are used, and baskets are placed in the stream so that the long axis is parallel to
stream flow. Samplers remain in the water for a period of 28 days (± 4 days) within the
sampling season (typically late August/early September). However, baskets may be required to
remain in the stream for 56 days (± 4 days) if the stream has a low velocity or is impounded.
Baskets are placed so that influences including bridges, culverts, channelized areas, slack water
areas, and eddies are avoided.
During removal of the rock baskets, they are approached from downstream to minimize
the kicking up of sediments and subsequent addition of organisms to the basket. Macrophytes,
algae, and debris are carefully removed from the surface of the basket. A 600-jim mesh net is
then placed against the substrate downstream of the basket and the basket is then quickly lifted
into the net. All contents of the net and the basket are then processed through a 600-jim sieve.
The basket and each rock are inspected to ensure the complete removal of all macroinvertebrates.
All of the sieved materials from each basket are then placed into a separate sample jar and
preserved with 95% ethanol and stream water to a final concentration of approximately 70%
ethanol in the field.
3.3.1.2 Laboratory Methods
The MDEP requires that all samples be handled by qualified personnel working under the
supervision of a professional aquatic biologist and that all taxonomy be performed by a person
who has experience in freshwater macroinvertebrate taxonomy. Valid samples must yield at
least 50 organisms each. The entire sample is sorted in small portions (1-2 tablespoons at a time)
until no more organisms can be found. Sorting is considered complete when no organisms are
found after 45 seconds of searching. While sorting, the sample is kept wet using water but does
not remain unpreserved for more than eight hours. All sorted organisms are then placed in a vial
containing a 70% alcohol and 5% glycerin solution. The detritus is returned to the sample jar
and preserved with 70% alcohol. Any samples used for regulatory purposes are kept for five
years. A qualified Biologist evaluates 10% of samples for sorting completeness.
3.3.1.2.1 Subsampling and Identification
If the mean number of organisms in a sampler is greater than 500, then subsampling may
be performed to yield at least 100 organisms per sampler. The MDEP does note that
subsampling will reduce sample richness by an amount that may affect the outcome when
performing linear discriminant analysis. This subsampling effect is taken into account by
biologists when making the final determination of classification attainment. If subsampling is
required, MDEP follows the methods outlined by Wrona et al (1982). First, sorted
macroinvertebrates are gently agitated in an Imhoff settling cone fitted with an aquarium bubbler
stone for two to five minutes. Then 25% of the sample is removed from the settling cone in five
aliquots using a settling cone and combined into one sample vial, insuring that the required 100
organisms have been obtained. It is important that the individual performing the subsampling
randomly dips from the cone and does not target specific organisms. Once the five aliquots are
5-6
-------
combined, the sample vial is then labeled indicating the fraction that the subsample represents.
An important precaution taken is assuring that large, dense organisms are not included in the
subsample because they are too heavy to be suspended for capture in the subsampler cone.
These organisms (e.g., crayfish, molluscs or caddisflies with stone cases) are counted separately.
The MDEP has tested the randomness of the sample distribution to conclude that five aliquots
can be combined into one sample (Elliott, 1977). After sorting, samples are identified by a
qualified taxonomist. The MDEP recommends that all macroinvertebrates be identified to
species; however, all numbers are adjusted to genus for use in metric calculations. In cases
where taxonomic expertise is lacking or when the specimen is too small or in poor condition, the
organism is identified to the lowest taxonomic level possible. Each taxonomist submitting data
to the MDEP is required to submit a reference collection of identified taxa and a list of
references used to identify samples. The reference collections are checked by a MDEP
taxonomist using the MDEP's master collection.
3.3.1.2.2 Chironomidae Identification and Subsampling
Chironomid midges are identified using slide mounts of the cleared head capsule and
body parts. For identification purposes, Euparol or Berlese mounting medium is preferred;
however, for permanent slide mounting, MDEP recommends that CMCP-9 be used (Wiederholm
1983).
For any sample containing less than 100 midges, all midges are identified to the lowest
possible taxonomic level. In samples containing 100-199 midges, a subsample containing 50%
of randomly selected specimens for identification, and in samples containing 200-499 midges,
25% of the specimens are randomly subsampled. The subsamples are identified to genus/species
level, and then the unsampled midges are examined for unusual or rare specimens. Those rare or
unusual specimens are also identified to genus/species level and kept separate from the
subsample. If a sample contains 500 or more midges, midges are grouped by genus, and a
random subsample of 100 organisms is selected from the remaining ungrouped midges. These
midges are identified to species level. If any rare or unusual specimens are found after
examination, they are identified to species level and kept separate from the subsample.
3.4 Data Management/Quality
The data generated (i.e., identified organisms) are entered into the database management
system ORACLE. ORACLE stores the taxonomic code table and all sampling event data,
computes analytical variables, and computes and reports the results of linear discriminant models
(Davies et al. 1999). All data are checked and verified following rigorous data entry and data
editing protocols. After entry into the database, all data are adjusted to the same taxonomic
resolution (genus) for comparison (Davies et al. 1995). All site data in ORACLE are also
georeferenced in ARCINFO so that spatial relationships can be studied.
3.5 Analysis of Biological Data (Information compiled from: Davies et al. 1995,
Davies et al. 1999, Davies and Tsomides 2002, MDEP 2003)
The MDEP uses linear discriminant analysis to assess the attainment of aquatic life
standards. A series of discriminant models based on macroinvertebrate metrics is used to divide
3-7
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observations among two or more predetermined classes (i.e., A, B, and C in Maine WQS). The
M.R.S.A. 38 Public Law Chapter 698: The Classification System for Maine Waters (State of
Maine 1985) outlines the system for assuring both the attainment of aquatic life and that the
objectives of maintaining the chemical, physical and biological integrity of waters are upheld by
the state. Minimum standards for dissolved oxygen, bacteria and aquatic life for each class, as
well as a detailed explanation of the classification system (Table 3-1), are described in this
document. The aquatic life standards are narratively established for attainment of each class and
are derived from measurable parameters. The results of the discriminant analysis are ultimately
used to assign each site to a class and determine ALUS.
The MDEP originally developed the linear discriminant models based on 145 rock basket
samples collected from across the state and representing a range of water quality during 1983-
1989. They recalibrated the models in 1998 using a much larger macroinvertebrate database
with a total of 376 sampling events (Davies et al. 1999). Quantifiable measures for each class
(A, B, C, and NA) were determined (Figure 3-3). The final step involved assigning each of the
376 sites in the database to one of four a priori groups using the quantifiable measures.
Linear discriminant analysis requires that a priori groups be established. A priori groups
consist of samples of "known" classification from which a predictive model can be developed to
characterize streams with unknown classifications (Davies et al. Draft). Based on Maine's Water
Quality Laws, MDEP chose four groups to which streams could be assigned: AA/A, B, C and
non-attainment (Figure 3-1). After testing multiple statistical modeling techniques (e.g., k-
means clustering, Two-Way Indicator Species Analysis, multivariate ordination), they decided
that the use of best professional judgment of expert aquatic biologists would be the most ideal
way to assign a priori groups. An explanation of the criteria biologists followed to establish the
a priori groups can be found in Appendix A. Once these groups were determined subjectively
and independently by three biologists, univariate and multivariate analysis of variance (ANOVA
and MANOVA, respectively) confirmed that the assigned groups were statistically distinct. To
determine variability in expert judgment assignments, a subset of data was assigned to a priori
groups by two non-MDEP biologists, yielding an average concurrence with MDEP biologists'
assignments of 80%. Furthermore, to check the model, MDEP chose 27 minimally disturbed
sites that were not originally used to build the model to determine the success of the model and
to assign them to Class A conditions. These sites had no known point sources and land uses
comprised 97% forested (3% logged), 2% crop and 1% residential/industrial/commercial.
Next, 25 biological community variables were ultimately identified from a list of 400
variables using stepwise discriminant analysis and iterative backward selection procedures to
best predict membership of an unknown stream sample to one of the four water quality classes
(A, B, C, and non-attainment). The 25 variables and the methods used to calculate them are
found in Table 3-2.
-------
Maine Tiered Uses Based on Measurable Ecological Values
Narrative Standard
Biological Value
Quantifiable Measures
Class A
natural
Class B
unimpaired, maintain
indigenous taxa
Class C -
maintain structure and
function
Taxonomic and numeric
equality; Presence of •
indicator taxa
Retention of taxa and
numbers; Absence of
hyperdominance; •
Presence of sensitive
taxa
Resistance;
Redundancy; Resilience;
Balanced distribution
Energy transfer;
Resource assimilation; •
Reproduction
Similarity, Richness,
"Abundance, Diversity, EPT,
Indicator taxa, Biotic Index
Community loss, Richness,
Abundance, Diversity,
>Equitability, Evenness, EPT,
Indicator taxa, Biotic Index
Richness, Diversity,
Equitability, Evenness
Trophic groups, Richness,
"Abundance, Community loss,
Fecundity, Colonization rate
Figure 3-3. Maine tiered uses based on measurable ecological values (modified from
Courtemanch 2003).
Linear discriminant functions were developed from the 25 quantitative macroinvertebrate
variables (Table 3-2). The discriminant functions determine the probability that a site belongs to
a given water quality class. Using a linear optimization algorithm to calculate the discriminant
function coefficients, multivariate space distance was minimized between sites within a class,
while the distance between classes was maximized. The linear discriminant model consists of
functions to compute an association value in the following form (MDEP 2003):
Z = C +WiXi +W2X2+ ..... WnXn.
Z = discriminant score
C = constant
W; = the coefficients or weights
X; = the predictor variable (metric) values
For each site, 25 quantitative metrics are calculated with the data from the three replicate
samples combined. Then, the discriminant function is calculated using one four-way model and
three two-way models. First, using only nine variables and calculated coefficients (Table 3-3),
the four-way model calculates the probability (range 0.0 - 1.0) that a site fits into each of the
three attainment classes (AA/A, B, or C), and the non-attainment (NA) class. The resultant
probabilities are then transformed and used as variables in the three two-way models (Table 3-4).
3-9
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Table 3-2. Methods for the calculation of variables and measures of community structure
used in linear discriminant models (from Davies and Tsomides, 2002).
Variable
Calculation Method
Total Mean
Abundance
Count all individuals in all replicate samples from one site and divide by
the number of replicates to yield the mean number of individuals per
sample.
Generic Richness
Count the number of different genera found in all replicates from one site.
Counting rules for Generic Richness:
a) All population counts at the species level are aggregated to the generic
level.
b) A family level identification that includes no more than one taxon
identified to the generic level is counted as a separate taxon in generic
richness counts.
c) A family level identification with more than one taxon identified to
generic level is not counted towards generic richness. Counts are divided
proportionately among the genera that are present.
d) Higher level taxonomic identifications (Phylum, Class, Order) are not
counted toward generic richness unless they are the only representative.
e) Pupae are ignored in all calculations.
Plecoptera Mean
Abundance
Count all individuals from the order Plecoptera in all replicate samplers
from one site and divide by the number of replicates to yield mean number
of Plecoptera individuals per basket.
Ephemeroptera
Mean Abundance
Count all individuals from the order Ephemeroptera in all replicate
samplers from one site and divide by the number of replicates to yield
mean number of Ephemeroptera individuals per basket.
Shannon-Wiener
Generic Diversity
(Shannon and
Weaver, 1963)
After adjusting all counts to genus following counting rules in Variable 2:
Where: C= 3.321928 (converts base 10 log to base 2)
N= Total abundance of individuals
n;= Total abundance of individuals in the ith taxon
Hilsenhoff Biotic
Index (Hilsenhoff,
1987)
N
Where: HBI= Hilsenhoff Biotic Index
n;= number of individuals in the ith taxon
al = tolerance value assigned to taxon i
N= total number of individuals in sample with tolerance values.
Relative
Chironomidae
Abundance
Calculate the mean number of individuals in the family Chironomidae,
following counting rules in Variable 4, and divide by total mean
abundance (Variable 1).
5-10
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8
9
10
11
12
13
14
15
16
Variable
Relative Diptera
Richness
Hydropsyche Mean
Abundance
Probability (A + B +
C) from First Stage
Model
Cheumatopsyche
Mean Abundance
EPT- Diptera
Richness Ratio
Relative Oligochaeta
Abundance
Probability (A+B)
from First Stage
Model
Perlidae Mean
Abundance (Family
Functional Group)
Tanypodinae Mean
Abundance (Family
Functional Group)
Calculation Method
Count the number of different genera from the order Diptera, following
counting rules in Variable 2, and divide by generic richness (Variable 2).
Count all individuals from the genus Hydropsyche in all replicate samplers
from one site, and divide by the number of replicates to yield mean number
of Hydropsyche individuals per basket.
Sum of probabilities for Classes A, B, and C from first stage model.
Count all individuals from the genus Cheumatopsyche in all replicate
samplers from one site and divide by the number of replicates to yield
mean number of Cheumatopsyche individuals per basket.
EPT Generic Richness (Variable 19) divided by the number of genera from
the order Diptera, following counting rules in Variable 2. If the number of
genera of Diptera in the sample is 0, a value of 1 is assigned to the
denominator.
Calculate the mean abundance in the class Oligochaeta, following counting
rules in Variable 4, and divide by total mean abundance (Variable 1).
Sum of probabilities for Classes A and B from the First Stage Model.
Count all individuals
from the family
Perlidae in all replicate
samplers from one site.
Divide by the number
of replicates to yield
mean number of
Perlidae per basket.
Count all individuals
from the subfamily
Tanypodinae in all
replicate samplers from
one site and divide by
the number of replicates
to yield mean number
of Tanypodinae per
basket.
Perlidae Functional Group
Acroneuria Neoperla
Agnetina Paragnetina
Attaneuria Perlesta
Beloneuria Perlinella
Eccoptura
Tanvpodinae Functional Group
Ablabesmyia Niltanypus
Clinotanypus Paramerina
Coelotanypus Pentaneura
Conchapelopia Procladius
Djalmabatista Psectrotanypus
Guttipelopia Rheopelopia
Husonimyia Tanypus
Labrundinia Telopelopia
Larsia Thienemannimyia
Meropelopia Trissopelopia
Natarsia Zavrelimyia
5-11
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17
18
19
20
21
22
23
24
Variable
Chironomini Mean
Abundance (Family
Functional Group)
Relative
Ephemeroptera
Abundance
EPT Generic
Richness
Variable Reserved*
Sum of Mean
Abundances of:
Dicrotendipes,
Microspectra,
Parachironomus and
Helobdella
Probability of Class
A from First Stage
Model
Relative Plecoptera
Richness
Variable Reserved*
Calculation Method
Count all individuals in
the tribe Chironomini in
all replicate samplers
from one site and divide
by the number of
replicates to yield the
mean number of
Chironomini per basket.
Chironomini Functional Group
Axarus Nlothauma
Chironomus Parachironomus
Cryptochironomus Paracladopelma
Cladopelma Paralauterborniella
Cryptotendipes Paratendipes
Demicriptochirono-mus Paenopsectra
Dicrotendipes Polypedilum
Endochironomus Pseudochironomus
Einfeldia Pagastiella
Gyptotendipes Robackia
Harnishchia Stelochomyia
Kiefferulus Stenochironomus
Lauterborniella Stictochironomus
Microchironomus Tribelos
Microtendipes Xenochironomus
Variable 4 divided by Variable 1 .
Count the number of different genera from the orders Ephemeroptera,
Plecoptera, and Trichoptera in all replicate samplers, according to counting
rules in Variable 2, generic richness.
Sum the abundances of the 4 genera and divide by the number of replicates
(as performed in Variable 4).
Probability of Class A from First Stage Model
Count number of genera of Order Plecoptera, following counting rules in
variable 2, and divide by generic richness (Variable 2).
5-12
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25
26
27
28
29
30
Variable
Sum of Mean
Abundances of
Cheumatopshyche,
Cricotopus,
Tanytarsus and
Ablabesmyia
Sum of Mean
Abundances of
Acroneuria and
Stenonema.
Variable Reserved*
Ratio of EP Generic
Richness
Variable Reserved*
Ratio of Class A
Indicator Taxa
Calculation Method
Sum of the number of individuals in each genus in all replicate baskets and
divide by the number of replicates (as performed in Variable 4).
Sum the number of individuals in each genus in all replicate baskets and
divide by the number of replicates (as performed in Variable 4).
Count the number of different genera from the orders Ephemeroptera (E),
and Plecoptera (P) in all replicate baskets, following counting rules in
Variable 2, and divide by 14 (maximum expected for Class A).
Count the number
of Class A
indicator taxa that
are present in the
sample and
divide by 7 (total
possible number).
Indicator Taxa: Class A
Brachycentrus (Trichoptera Brachycentridae)
Serratella (Ephemeroptera: Ephemerellidae)
Leucrocuta (Ephemeroptera: Heptageniidae)
Glossosoma (Trichoptera: Glossosomatidae)
Paragnetina (Plecoptera: Perlidae)
Eurylophella (Ephemeroptera: Ephemerellidae)
Psilotreta (Trichoptera: Ondontoceridae)
*These variable numbers are not used in discriminant models.
5-13
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Table 3-3. Coefficients for the First Stage Model (from MDEP 2003).
Variable #
Constant
1
2
3
4
5
6
7
8
9
Coefficients
Transformation
In (value + 0.001)
In (value + 0.001)
In (value + 0.001)
In (value + 0.001)
Class A
-99.95508
10.77061
-0.38619
0.23940
-0.59970
21.22732
8.01620
-11.70298
70.77937
-0.00535
Class B
-105.70948
11.46981
-0.43340
0.03946
-0.55500
20.91256
9.12163
-11.52650
71.09637
-0.00398
Class C
-112.67581
11.80888
-0.50051
-0.60923
-0.67722
21.07602
10.31492
-11.49414
72.46514
-0.00152
Nonattainment
-107.74283
11.26793
-0.48822
-0.95480
-1.79032
19.46547
10.72746
-11.66371
70.22517
0.00007
Table 3-4. Coefficients for the Final Classification Models (AA/A, B, and C) (MDEP 2003).
Class C or Better Model
Variable #
Constant
10
11
12
13
Coefficients
Transformation
Arcsin
In (value + 0.001)
Square Root
In (value + 0.001)
Class A-B-C
-25.70020
19.98470
-0.26001
5.57672
-2.33229
Nonattainment
-8.55844
3.36032
-0.43781
5.92732
-1.89945
Class B or Better Model
Variable #
Constant
14
15
16
17
18
19
21
Coefficients
Transformation
Arcsin
In (value + 0.001)
In (value + 0.001)
In (value + 0.001)
In (value + 0.001)
Class A-B
-17.81016
12.04826
-1.11091
-0.10582
0.17813
4.03202
0.87400
-0.69360
Nonattainment
-6.93836
3.63707
-1.03934
0.01978
0.10825
-1.14508
0.63310
-0.53194
5-14
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Class A Model
Variable #
Constant
22
23
25
26
28
30
Coefficients
Transformation
Arcsin
In (value + 0.001)
In (value + 0.001)
Class A-B-C
-9.59254
8.34341
3.78999
0.53110
-0.55838
12.32529
6.94828
Nonattainment
-4.08552
1.52080
4.27447
0.77851
-0.51448
9.81592
-0.67475
The next step is to use three two-way models to test the probability of a site belonging to
class AA/A, B, or C. These models distinguish between a given class plus any higher classes as
a group and any lower classes as a group (i.e., Classes AA/A + B + C vs NA; Classes AA/A + B
vs Class C + NA; Class AA/A vs Classes B + C + NA). The three models are applied using the
equations in Table 3-4 (MDEP 2003, Davies and Tsomides 2002).
The discriminant scores (Z) are known as the Mahalanobis Distances (MDEP 2003)
where:
Mahalanobis Distance = Zt(sample x) = gi (x,f) + g2 (f)
And where:
Zt = discriminant score for sample x, class t
gi (x,f) = (x- mf) S"1 (s-mf)
g2 (t) = -2 loge (qf) = 0 (if prior probabilities are equal)
x = a vector containing all the values of all the variables for a given linear discriminant
function, for a given sample, of class t
mf = a vector, as for x, but containing the means of all predictor variables in the given linear
discriminant function, for the given sample, of class t
S = pooled covariance matrix (the variance of the multivariate observation)
qf = value of the prior probability that a given sample is Class A, B, C, or NA
The probability (association value) of a sample x belonging to a particular class t is:
5-15
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± t V-V - NA
I [e[-°5
A
Where:
Pf(x) = the probability that sample x belongs to class t (for Classes AA/A, B, C, NA)
e = the exponential function
-0.5 = a standardized constant from the normal distribution
Zf = the discriminant score or Mahalanobis Distance for class t (Classes AA/A, B, C, NA)
Once the probability that a site belongs to a certain class is calculated, the MDEP follows
a process to determine whether the site attains at least that particular class (Davies and Tsomides
2002, MDEP 2003). In order to determine whether a site attains at least Class C or is in non-
attainment, the association value (Zc) calculated using the 'C or better model' needs to be used.
If the association value is greater than 0.6, then the sample attains to Class C or higher, but if it is
less than 0.4 then the site is in non-attainment. If a site falls within 0.4 - 0.6, then best
professional judgment is used to determine if the site belongs in Class C, is in non-attainment, or
is indeterminate of Class C. For any site found to be indeterminate, additional monitoring may
be needed in order to make a decision.
Those samples that do attain to at least Class C are then tested for Class B attainment
using the association values (ZB) from the 'B or better model'. If the association value is greater
than 0.6 then the sites are at the minimum attaining to Class B status. Those values below 0.4
are now considered to be sites that attain to Class C. If a value falls between 0.4 - 0.6 then it is
indeterminate of Class B and may require additional monitoring to determine to which
attainment class the site belongs.
When the association value for a site is 0.6 or greater using the Class B or better model, it
is then tested using the 'A or better model'. If the association value (ZA) is 0.6 or greater, then
the class attains to A. If the value is 0.4 or less, then the class attains to Class B. If the value is
between 0.4 - 0.6, the finding is indeterminate of Class A and additional sampling may be
required. Figure 3-4 graphically shows the process for calculating model variables and
association values using linear discriminant models and Figure 3-5 shows how the attainment
classes are determined using association values (MDEP 2003).
After a site is placed into an aquatic use attainment class, a provision in MDEP regulation
Chapter 579 (MDEP 2003) allows for professional judgment to make an adjustment to the
evaluation. Any adjustment may be made using analytical, biological, and habitat data.
Professional judgment also may be employed when the condition of the stream does not allow
for the accurate use of the linear discriminant models. Such factors may include habitat
influences (e.g., lake outlets, impounded waters, substrate characteristics, tidal waters), sampling
issues (e.g., disturbed samples, unusual taxa assemblages, human error in sampling), or
analytical and sample processing issues (e.g., subsample vs. whole sample analysis or human
error in processing) (Davies and Tsomides 2002).
If a water body falls into a lower class than its assigned statutory class after a MDEP
biologist determines that adjustment is not needed, then the site is determined to be in non-
attainment of its statutory class. When a site is found to be in non-attainment of its legally
5-16
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defined class, then certain actions must be taken that include: 1) the notification of other
programs (e.g., Licensing or Land Use Regulation); 2) the listing of the stream in question on the
303(d) list of impaired water bodies; and 3) the development of a total maximum daily load
(TDML) for pollutants.
If a water body falls into a higher class than its assigned statutory class, then the higher
aquatic life conditions must be maintained if the finding confirmed under critical water quality
conditions. This finding also requires certain actions to ensure the findings were accurate and
include: 1) the confirmation of the finding by resampling; 2) the confirmation that higher aquatic
life quality exists at the maximal pollution loads; and 3) MDEP can propose that the water body
be upgraded if dissolved oxygen, bacteria and habitat standards also adhere to the next higher
class (MDEP 2002). Furthermore, data collected from two or more sampling events over
different seasons must be evaluated to determine if a site has improved in quality and moved into
the next higher statutory class.
All results and classification attainment decisions made by MDEP must be reported in the
State of Maine Water Quality Assessment Report that is required under Section 305(b) of the
CWA. If a water body is found to be in non-attainment of its statutory classification, then it is
placed on Maine's list of impaired waters as required in Section 303(d) of the CWA.
3.6 Literature Cited
Courtemanch, D. L. 2003. RFC 101: Development of Reference Conditions for Management
Classes. National Biological Assessment and Criteria Workshop: Advancing State and
Tribal Programs. Coeur d'Alene, ID. March 31-April 3, 2003.
http://www.epa.gov/waterscience/biocriteria/modules/rfcl01-07-maine.pdf
Davies, S.P., L. Tsomides, D.L. Courtemanch, andF. Drummond. 1995. Maine Biological
Monitoring and Biocriteria Development Program, 2nd edition. Maine Department of
Environmental Protection, Augusta, ME.
Davies, S.P., L. Tsomides; J.L. DiFranco and D.L. Courtemanch. 1999. Biomonitoring
Retrospective: Fifteen Year Summary for Maine Rivers and Streams. Maine Department of
Environmental Protection, Augusta, ME.
http://www.state.me.us/dep/blwq/docmonitoring/biomonitoring/biorep2000.htm
Davies, S.P. and L. Tsomides. 2002. Methods for the Biological Sampling and Analysis of
Maine's Rivers and Streams. Maine Department of Environmental Protection, Augusta, ME.
http://www.state.me.us/dep/blwq/docmonitoring/fmlmethl.pdf
Davies, S.P., L. Tsomides, D.L. Courtemanch, and F. Drummond. Draft. Stream Biological
Monitoring and Numeric Criteria Development in Maine. Maine Department of
Environmental Protection, Augusta, ME.
Elliot, J.M. 1977. Some Methods for the Statistical Analysis of Samples of Benthic
Macroinvertebrates. Freshwater Biological Association., Science Publication No. 25.
5-17
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Computer calculates model variables (Varl - VarSO)
using taxa counts from a sample event using
procedures described in Table 3-2.
First Stage Linear Discriminant Model (LDM)
(4-way model: A vs. B vs. C vs. NA)
1. Model calculates Discriminant Score using Varl - Var9.
2. Model uses Discriminant Score to calculate the following
Association Values1:
• probability Class AA/A (pAl)
• probability Class B (pBl)
• probability Class C (pCl)
• probability Nonattainment (pNAl)
1.1 "C or Better" Second Stage LDM
(2-way model: A, B, or C vs. NA)
1. .Model calculates Discriminant Score
using VarlO (pAl+pBl+pCl) and
Varll - Varl 3.
2. Model uses Discriminant Score to
calculate the following Association
Values:
• probability C or better (pABC)
• probability NA (pNA)
1.2 "B or Better" Second Stage LDM
(2-way model: A or B vs. C or NA)
1. .Model calculates Discriminant Score
using Varl4 (pAl+pBl) and VarlS -
Var21.
2. .Model uses Discriminant Score to
calculate the following Association
Values1:
• probability B or better (pAB)
• probability C or NA (pCNA)
1.3 "A" Second Stage LDM
(2-way model: A vs. B, C, or NA)
1. .Model calculates Discriminant Score1
using Var22 (pAl) and Var23 — VarSO.
2. .Model uses Discriminant Score to
calculate the following Association
Values1:
• probability AA/A (pA)
• probability B, C, or NA
(pBCNA)
Figure 3-4. Process of calculating model variables and association values using linear discriminant models (taken from MDEP
2003).
5-18
-------
Is the sample appropriate for Linear Discriminant Model?
1
I
(" Use BPJ J
r
Is the sample Class C or better?
r At least C J
At least C U Indeterminat
/\ e
NA
( NA )
I
Is the sample Class B or better?
At least B V Indeterminate
Is the sample Class A?
A V Indeterminat
e
B
A £
Figure 3-5. Process for determining attainment class using association values (modified
from MDEP 2003).
5-19
-------
Klemm, D.J., P.A. Lewis, F. Fulk, and J.M. Lazorchak. 1990. Macroinvertebrate
and Field Methods for Evaluating the Biological Integrity of Surface Water.
EPA/600/4-90/030. U.S. Environmental Protection Agency, Cincinnati, OH.
(MDEP) Maine Department of Environmental Protection. 2002. Board of
Environmental Protection Meeting, Frequently Asked Questions. Unpublished
material.
MDEP. 2003. Classification Attainment Evaluation Using Biological Criteria for Rivers and
Streams. Rules of Maine State Government Agencies, 06 096 Chapter 579.
http://www.maine.gov/sos/cec/rcn/apa/06/096/096c579.doc
State of Maine. 1985. Maine Laws Ch. 698 § 15 (in part). An Act to Amend the
Classification System for Maine Waters and Change the Classifications of
Certain Waters. Augusta, Maine.
Wiederholm, T. 1983. Chironomidae of the Holarctic Region. Entomologica
Scandinavica, Suppl. No. 19.
Wrona, F.J., J.M. Gulp and R.W. Davies. 1982. Macroinvertebrate subsampling:
a simplified apparatus and approach. Canadian Journal of Fisheries and
Aquatic Science 39:1051-1054.
3.7 Resources
Maine Department of Environmental Protection, Biological Monitoring Web Page:
http://www.state.me.us/dep/blwq/docmonitoring/biomonitoring/index.htm
Maine Revised Statutes Annotate. Title 38, Chapter 3: Protection and Improvement of Waters,
Sections 464 and 465. http://janus.state.me.us/legis/statutes/38/title38ch3secO.html
U.S. EPA. Biological Indicators of Watershed Health, Maine Webpage:
http://www.epa.gov/bioindicators/html/state/me-bio.html
U.S. EPA. 2002. Summary of Biological Assessment Programs and Biocriteria
Development for States, Tribes, Territories, and Interstate Commissions:
Streams and Wadeable Rivers. EPA-822-R-02-048. U.S. Environmental
Protection Agency.
5-20
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4 MASSACHUSETTS
This document was prepared using documents written by the
Massachusetts Department of Environmental Protection. Any
questions concerning bioassessment methods should be directed to:
Arthur Johnson, Environmental Monitoring Coordinator
Massachusetts Department of Environmental Protection (MA
DEP)
627 Main Street
Worcester, MA 01608
Phone: (508) 767-2873; Fax (508) 791-4131
Email: Arthur.Johnson@state.ma.us
4.1 Introduction
As required by the CWA, MA DEP submits a biennial 305(b) report that describes the
status of the state's water resources with respect to the classes defined by the Massachusetts
Surface Water Quality Standards. The Massachusetts Surface Water Quality Standards assign
support classes according to intended use, which include aquatic life, fish and shellfish
consumption, drinking water supply, primary recreational contact, and secondary recreational
contact (314 CMR 4.00 2000, Table 4-1). These standards were set to account for the most
severe hydrologic conditions. In rivers, the standards must apply to waters at or above the seven-
day ten-year flow statistic (7Q10). In regulated water systems, the standards must apply to the
lowest discharge that meets or exceeds criteria 99% of the time on a yearly basis and any
alternatives must be approved by the MA DEP Commissioner or the entity controlling flow (MA
DEP 2003).
The ALUS criteria of the standards require that a suitable habitat be provided, "to sustain
a native, naturally diverse community of aquatic flora and fauna" (MA DEP 2003). For
assessment purposes each attainment class is composed of two sub-classes, the cold water fishery
(capable of sustaining a year-round population of cold water aquatic organisms) and the warm
water fishery (not capable of sustaining a year-round population of cold water organisms). Each
stream that is assessed is designated as fully supporting, partially supporting, or non-supporting
for its ALU class. Sites that are designated as "fully supporting" for a particular attainment class
also may be given a threatened status if the stream is in danger of becoming polluted within a
two-year period. Sites with too little or no data are not assessed.
The MA DEP uses biocriteria to aid in the assessment of the ALUS of surface waters.
Macroinvertebrate, periphyton, and fish data are used by Massachusetts State Biologists to aid in
determining ALUS attainment. Furthermore, MA DEP combines the biological data with habitat
evaluations, toxicological and chemical data, and other environmental variables in order to make
a final ALUS decision.
4-1
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Table 4-1. Massachusetts attainment classes with management strategy and narrative
biological and habitat criteria as stated in 314 CMR 4.00 (2000).
Management
Biological and Habitat Narrative
Criteria
These waters are designated as a source of public
water supply. To the extent compatible with this use
they shall be an excellent habitat for fish, other
aquatic life and wildlife, and suitable for primary
and secondary contact recreation. These waters shall
have excellent aesthetic value. These waters are
designated for protection as Outstanding Resource
Waters under 314 CMR 4.04(3).
Excellent habitat for fish, other
aquatic life and wildlife, supporting
normal species diversity, successful
migration, reproductive functions or
growth of aquatic organisms.
B
These waters are designated as a habitat for fish,
other aquatic life, and wildlife, and for primary and
secondary contact recreation. Where designated they
shall be suitable as a source of public water supply
with appropriate treatment. They shall be suitable
for irrigation and other agricultural uses and for
compatible industrial cooling and process uses.
These waters shall have consistently good aesthetic
value.
These waters are designated as a habitat for fish,
other aquatic life and wildlife, and for secondary
contact recreation. These waters shall be suitable for
the irrigation of crops used for consumption after
cooking and for compatible industrial cooling and
process uses. These waters shall have good aesthetic
value.
These waters are designated as a
habitat for fish, other aquatic life
and wildlife, protect normal species
diversity, successful migration,
reproductive functions or growth of
aquatic organisms.
4.2
Key Elements of the Biological Assessment Approach
4.2.1 Index Period and/or Temporal Conditions
Originally, in order to represent the "worst-case" scenario, all sampling events were
performed during low-flow, dry-weather, high-stress conditions. However, when MA DEP
began to consider the impact of non-point source pollution, they altered the schedule to include a
larger range of weather conditions. Typically, the sampling season is July through September
and this sampling schedule is consistent from year to year so that data collected from surveys can
be compared to make historical inferences. This time frame also coincides with streams
returning to base flow conditions and occurs after the spawning season for most fish species
present within the state.
4.2.2 Monitoring Program Survey Approach
The protocol of the MA DEP biomonitoring program divides the state into 27 major
watersheds and coastal drainage areas that are sampled using a five-year basin rotation strategy
4-2
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(MA DEP 2003, Figure 4-1). Within each basin, the first year of the program involves
reconnaissance of basins and research to identify available data and/or information gaps. During
the second year, streams are sampled and data are collected according to a MA DEP Quality
Assurance Project Plan (QAPP). These sites are targeted based on known or suspected water
quality degradation. In the third year, the stream assessments are made and published in State
documents and/or 305(b) reports. Over the next two years, other watershed management issues
are targeted (e.g., TMDL calculations, permit issuance, targeting of problem watersheds,
outreach program implementation). In order to determine ALUS, MA DEP samples
approximately 75 streams per year.
4.2.3 Natural Classification of Water Bodies
Massachusetts contains three Level III Ecoregions: the Northeastern Highlands, the
Northeastern Coastal Zone, and the Atlantic Coastal Pine Barrens (Griffith et al. 1994). The
Northeastern Highlands contains seven Level IV ecoregions (Taconic Mountains, Western New
England Marble Valleys, Green Mountain/Berkshire Highlands, Lower Berkshire Hills,
Berkshire Transition, Vermont Piedmont, and Worcester/Monadnock Plateau). The Northeastern
Coastal Zone contains five Level IV ecoregions (Connecticut Valley, Lower Worcester
Plateau/Eastern Connecticut Upland, Southern New England Coastal Plains and Hills, Boston
Basin, and Narraganset/Bristol Lowland). The Atlantic Coastal Pine Barrens contains one Level
IV ecoregion (Cape Cod/Long Island). For sampling purposes, the state is assessed on a basin-
level scale (Figure 4-1), which may incorporate more than one Level IV ecoregion (Figure 4-2).
Cold water and warm water streams are assessed as separate categories.
4.2.4 Indicator Assemblages
Biological, toxicological and chemical data are collected and used to make ALUS
decisions based on a "weight of evidence" approach. Biological indicators include assessments
of the macroinvertebrate, fish, and algal communities. An index based on the RBP II and III
(Plafkin et al. 1989) is used to assess the macroinvertebrate assemblage. The overall structure
and condition of the fish assemblage is assessed using some measurements from the RBP V
(Plafkin et al. 1989). Water quality condition is also assessed using algal measurements (i.e.,
Chlorophyll a concentration, percent cover of green algae, and biomass).
4.2.5 Reference Condition (Arthur Johnson, Personal Communication)
Reference sites are identified by examining criteria and by using the best professional judgment
of MA DEP Biologists. Sites that are described as "least impacted" have minimal or no potential
to receive point or non-point source pollution and lack land use patterns that would degrade
water or habitat quality. Maps and field reconnaissance are both used to locate unique reference
sites when RPBs are used to assess a site. Multiple reference sites often exist for a study and
occasionally reference sites of adjoining watersheds of the study site may be used. Sites are only
considered reference for streams of similar elevation and drainage area.
4-3
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Five Year Basin Cycle
SCHEDULE OF FIVE-YEAR BASIN CYCLE
2002
1
5
4
3
2
2003
2
1
5
4
3
2004
3
2
1
5
4
2005
4
3
2
1
5
2006
5
4
3
2
1
2007
1
5
4
3
2
Team functions during each phase(year) of the cycle:
Year 1: Outreach
Year 2: Research
Year 3: Assessment
Year 4: Planning/Implementation
Year 5: Evaluation
0 7 14 21 28 35 Miles
Figure 4-1. Massachusetts 5-Year Basin Rotation Strategy (taken from the Massachusetts
Department of Environmental Protection website
www. mass.£ov/dep/brp/wni/files/cvclemap6. jps).
4.3 Field and Laboratory Protocols
4.3.1 Macroinvertebrate Protocols (Taken from Nuzzo 2003)
4.3.1.1 Field Methods
The MA DEP uses one of three different methods to collect macroinvertebrates,
depending on the depth and substrate of the stream. Kick sampling is the most commonly used
method and is designed for use in wadeable streams that have coarse substrates, rock baskets are
used in streams that are not wadeable or have fine substrates and for studies that require
quantitative measurements, and Hester-Dendy multi-plate samplers are used in deep rivers. Each
of the methods requires a habitat assessment to accompany the macroinvertebrate data to identify
problematic areas and habitat destruction or loss. The MA DEP uses a visual-based rapid habitat
assessment method (as described in Barbour et al. 1999) that includes ten habitat categories that
are rated from 0 (lowest) to 20 (optimal).
4-4
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Level 111 Mi'l IV Kcorcgiuns of Massachusetts, Rhode I - l.
-------
4.3.1.1.1 Kick Sampling
Kick sampling is used for routine biomonitoring in wadeable streams with coarse
substrates and riffle habitats. A 100-meter reach is chosen to represent the best habitat for the
particular sample area. Then, a 0.46-m x 0.46-m, 500-|im mesh kick net is placed into the riffle
and pressed firmly against the bottom so that the net is facing upstream. A biologist then kicks
upstream in an area approximately the same size as the kick net, allowing material to be caught
in the net. Following a kick (minimally 30 seconds), the net is examined, any
macroinvertebrates residing on large debris are rinsed off into the net, and the excess debris is
then placed back into the stream. In streams where the riffles within the reach are inadequate to
allow for a 2-m2 composite, other productive habitats are sampled by jabs into snags and/or by
rubbing substrates. The contents of the net are then emptied into a 2-L wide-mouth leak-proof
Nalgene bottle. Ten kick samples (approximately equaling 2-m2) are collected at a site and
composited into one bottle. The contents are preserved with enough denatured 100% reagent
alcohol (5% methanol, 5% isopropanol, 90% ethanol) to cover the residue. If preservative is not
added to the sample, it must be placed on ice and processed within 48 hours of collection.
4.3.1.1.2 Rock Basket Sampling
When kick sampling is deemed inappropriate for stream biomonitoring, rock baskets are
used to collect benthic macroinvertebrates. Situations where rock baskets may be employed
include planned statistical treatments of benthos data requiring quantitative collection methods,
water depth too great for kick sampling, and substrate which is too fine for kick sampling at one
or more sites used for comparison studies. The MA DEP uses rock baskets that are filled with
roofing stone. Each basket is approximately the same weight and uses 2.5 -7.5 cm sized rocks.
Three baskets are placed separately at the bottom of the stream in a riffle for six to eight weeks
(in a current velocity of 15 - 76 cm/s). Upon retrieval, a kick-net is pressed tightly against the
stream bed along the edge of the basket's downstream edge. The basket is then lifted onto the
net. If the removal is difficult due to water depth, a cover is draped over the basket
(Courtemanch 1984). After removal, each basket is placed into a separate large bucket or tub of
water and then opened. All of the contents are emptied into the bucket and each rock is rinsed
and set aside. After all of the rocks have been rinsed, the contents of each bucket are run through
a #30 mesh (600 jim) and then placed into a sample container and preserved with denatured
100% reagent alcohol. If the sample is to be processed within 48 hours, the sample is put on ice
without preservative.
4.3.1.1.3 Hester-Dendy Multi-plate Sampling
Hester-Dendy multi-plate samplers are used in deep rivers or where kick sampling is
inappropriate and rock baskets are impractical. The MA DEP uses round-plate samplers as
specified by U.S. EPA (Klemm et al. 1990). The samplers are placed in deep waters and
tethered to an anchored float so that the sampler is suspended 1 m below the surface. Samplers
placed in shallow water are mounted to a patio block or 4-in cinder blocks. In order for
comparable data to be collected, each sampler is placed in a riffle of a similar velocity (within
the range of 15 and 76 cm/s). Three Hester-Dendy multi-plate samplers are independently
deployed in a stream at the same time for six to eight weeks. When they are retrieved, samplers
4-6
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are completely surrounded with a 500-jim mesh net, a plastic bag, or a 2-L wide-mouth jar and
pulled from the water. Enough water is placed in the 2-L wide-mouth jar to completely cover
each sampler and then it is tightly capped and placed on ice and labeled. All multi-plate
samplers are removed, placed in separate bottles in a refrigerator, and processed within 48 hours.
After processing, samples are preserved with 100% reagent ethanol.
4.3.1.2 Laboratory Methods (Nuzzo 2003)
4.3.1.2.1 Processing of Kick Net and Rock Basket Samples
The MA DEP uses RBPs to process the macroinvertebrate data collected using kick nets
or rock baskets (RBP II and RBP III; Plafkin et al. 1989) to obtain 100 organisms in the
subsample. Each sample to be sorted is first run through a 600-jim mesh sieve and held over a
waste collection vessel to collect the decanted alcohol preservative, then rinsed three times with
enough water so that the sample is washed free of any preservative. Following rinsing, the
sample is left to drain in the sieve for one minute and then the sample is spread evenly across a
gridded pan that contains 25 squares 6-cm x 6-cm. Multiple pans are used to distribute the
material evenly and to reduce the density of organisms per grid to a workable number (e.g., pan
A contains grids 1-25, and pan B contains grids 26 - 50). Sufficient water is then added to
distribute the material evenly across the pans. Next, the sorter chooses random numbers within
the range of the sum of total squares used to hold the sample. Once a random grid square is
chosen, only the material from that square is removed and it is placed into a petri dish that has
been divided into quarters. One of the quadrants inside the petri dish is randomly selected and
all organisms are sorted from that portion. The remaining debris in the petri dish is discarded
into the waste collection vessel. The process is then repeated, so that complete quadrants are
sorted until at least 100 organisms are obtained in the sample or until sorting yields no fewer
than 90 organisms and the spread between the highest and lowest count among subsamples being
compared is not greater than ±10%. If a problem arises from the original sample because the
primary data are questioned, an additional 100-200 organisms are sorted, stored separately and
labeled as "subsample extras". Furthermore, if any specimens in good condition are
encountered, they may be kept separately from the subsample and stored in separate vials as
voucher specimens.
4.3.1.2.2 Processing of Hester-Dendy Multi-plate Samples
Hester-Dendy multi-plate sampling of the macroinvertebrate assemblage uses a different
protocol because all processing must take place within 48 hours of collection. After removal
from the refrigerator, the entire multi-plate sampler and the water in which it is stored are
transferred into a large pan. The sample container is then rinsed into the pan three times with
small volumes of water to ensure that no organisms remain attached to the inside of the
container. The multi-plate is then disassembled and all pieces are rinsed. The sample is then
poured through a #30 mesh sieve, and the resultant debris is placed into ajar and preserved with
denatured 70% ethanol or sorted immediately as required by the study design. Normally all
macroinvertebrates recovered are identified; however, if the macroinvertebrate density is high,
only a subsample is identified.
4-7
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4.3.1.2.3 Taxonomic Identification
A taxonomist identifies macroinvertebrates to genus or species level if the RBP III
protocol is used. Identification may be to the family or order level if the RBP II protocol is being
followed, or if the organism is in poor condition, immature, or not identified in an available key.
MA DEP uses a number of keys to identify macroinvertebrates (Klemm 1985; Kathman and
Brinkhurst 1998; Jokinen 1983; Smith 1986; 1987; 1988; Merritt and Cummins 1996; and
Peckarsky et al. 1990). After identification, specimens are placed in labeled glass vials and
preserved with denatured 70% ethanol.
4.3.1.2.4 Oligochaeta and Chironomidae Identification
Chironomid midges and oligochaetes are identified using slide mounts under a compound
microscope using permanent or semi-permanent mounting medium. A 3-in x 1-in microscope
slide is situated with the label end facing left. Three worms or midges are then placed side-by-
side with heads towards the top. A sufficient amount of CMC-10 is then added to the specimens
to cover them and surround them. Worms are uncoiled and placed on their sides using forceps.
The heads of the midges are separated from the body using an insect pin and forceps and then
they are oriented so that their bodies are on their side and the heads are placed with the ventral
side up. A #1 cover slip is then added and gentle pressure is applied to ensure that all air bubbles
have been removed and so that the mandibles of midges are opened. CMC-10 is added to the
edge of the cover slips and the slides are dried for at least one week horizontally before
placement into a storage box. Identification by taxonomists are made using the keys by Kathman
and Brinkhurst (1998), Bode (1990), Coffman and Ferrington (1996), and Wiederholm (1983,
1986).
4.3.2 Periphyton Protocols (Beskenis 2002, Draft 2003; Barbour et al. 1999)
4.3.2.1 Field Methods
MA DEP biologists evaluate stream condition using assessments of the periphyton
assemblage above and below point and non-point sources or compared to a reference stream to
look for toxicity issues, nutrient impacts and habitat alterations. The data from the periphyton
sampling is used to evaluate if either the aesthetics or ALU is affected. MA DEP identifies both
diatoms and soft algae to the genus level, and they incorporate other algal measurements:
biomass (AFDM), chlorophyll a determination, and percent coverage.
4.3.2.1.1 Algal Abundance and Identification
Within the same 100-m reach used to collect macroinvertebrate samples (see section
4.3.1), benthic algal samples are collected using the RBP Single Habitat Approach from riffles
with a current of 10-50 cm/s. All samples are scraped from either natural or artificial substrates
following U.S. EPA's RBPs (Barbour et al. 1999). Substrate (preferably cobble) within the
stream reach is scraped with a knife and the detached algae are then washed into a labeled glass
vial using stream water. All vials collected from a site are tightly closed and placed into a large
plastic jar containing stream water to ensure a regular temperature is maintained. Then the
4-8
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samples are transported back to the laboratory to be stored in the refrigerator until identified and
counted. If a sample is held for an extended time before analysis the sample is preserved with
M3 fixative.
4.3.2.1.2 Biomass
MA DEP uses artificial substrates to collect algae data that determine periphyton biomass
and chlorophyll a measurements (APHA 1992). The artificial substrate apparatus is constructed
from a wooden tray attached to a cinder block with nylon twine. Slides are placed vertically in
the wooden tray and the apparatus is placed into the water at a depth of at least 0.25 m and less
than 1 m for a period of approximately three weeks. Any duplicate apparatuses are placed at
comparable light and flow regimes.
4.3.2.1.3 Chlorophyll a
Chlorophyll a samples are typically measured using two slides from trays that have been
deployed into a stream for three weeks to measure biomass. The two slides removed from the
tray are then placed on ice, brought back to the lab and either filtered and ground or frozen until
analysis (Beskenis Draft 2003). An alternative method to collect periphyton chlorophyll a is to
scrape a known area of rocks clean. Then, the material is rinsed into a bottle with water and a
subsample is removed for chlorophyll a analysis. Samples are typically processed immediately
upon arrival and must be analyzed within 21 days of first filtering or freezing.
4.3.2.1.4 Percent coverage calculation (Beskenis 2002)
Percent algal cover is determined in the riffle zones and is usually determined visually by
the biologist conducting the on-site survey. The surveyor will also note whether the algal
composition is microalgae or macroalgae. If a stream consists of greater than 50% macroalgae,
it may be considered to be threatened based upon review of other data including, if available, DO
measurements and impacts on the macroinvertebrate assemblage.
MA DEP also uses the point-intercept method to determine percent algal cover. For this
method, a bottom viewer constructed by cutting a hole in the bottom of a bucket greater than 0.5
m wide) and attaching a piece of clear acrylic material to the bottom with silicon caulk is used to
assess the algal cover. The viewing bucket has 50 evenly spaced dots forming a grid on the clear
acrylic bottom. Three transects are then laid out across areas where benthic algae are observed,
insuring that the left and right banks and the middle are covered. The viewer is then placed into
the stream and the number of dots covering the macroalgae are counted and recorded. A
subsample of the macroalgae is collected for identification. Then, the process is repeated to
count the number of rocks that contain substrata with microalgae on them. The microalgal
composition is assessed to determine if it is composed of diatomaceous, green algae, or blue-
green algae and a subsample is collected for identification. If an algal mat exists, the thickness is
determined in the field using a ruler and a score is given to describe the mat as a function of
thickness:
4-9
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Score
0 Substrate is rough with no visible evidence of microalgae
0.5 Substrate is slimy and no visible accumulation of microalgae is present
1 A thin layer of microalgae is present
2 Accumulation of a microalgae mat from 0.5-1 mm thick is present
3 The microalgae mat is 1- 5 mm thick
4 The mat is 5 mm - 2 cm thick
5 The mat is > 2 cm
Abundance is calculated by counting the number of organisms of each genus in each field
view. If less than one cell per field is counted, the genus is considered rare (R). If at least one
cell of a genus (but fewer than five cells) is counted per field, the genus is common (C). If
between five and 25 cells are counted per field then the genus is very common (VC). A genus is
abundant (A) if greater than 25 cells per field are present and very abundant (VA) if the cells are
too abundant to count.
4.3.2.1.5 Biomass Determination
When the artificial substrate sampling device is retrieved, the wooden trays and slides are
removed from the cinder blocks and placed in plastic bags on ice. The two outside slides are
discarded and a random number generator is used to choose two slides to be analyzed for
chlorophyll a (See chlorophyll a determination). The remaining four slides are used to measure
biomass as AFDM. After removal from the tray the slides are first air-dried. China crucibles are
cleaned and dried at 105°C and then cooled in a desiccator and weighed. Crucibles are returned
to the oven for one hour, cooled and re-weighed until there is less than a 1% change in weight for
each of the crucibles. After the slides are removed from the muffle furnace, one slide is retained
for the archive and three of the slides are used as replicates and scraped into the pre-cleaned and
pre-weighed china crucibles. A few drops of water are added to the crucibles and then they are
placed into the combustion oven at 500°C for one hour (Thermolyne Sybron furnatrol 11, type
13300). After the oven cools to 200°C, the samples are removed and placed in a dessicator
before water is added for re-hydration. After the sample is re-hydrated, it is dried to a constant
weight at 105°C and then the final weight is recorded. To calculate productivity, the following
equation is used:
r> (mg of AFDM per slide )
~~ ~A
P= net productivity (mg ash-free weight/m2/day)
t= exposure time (days)
A= area of slide (m2)
The mean weight from the slides is calculated and reported as dry weight and ash-free dry
weight/m2 using the formula:
/ 2 Mean AFDM in g
£/m ~ 0.00375m2
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To calculate the Autotrophic Index (Biggs 1996):
A T Biomass (ash free weight of organic matter in mg I m)
Chlorophyll a in mg I m
AI values range from 50 - 200. Larger values indicate a greater number of heterotrophic (fungi,
bacteria) than autotrophic (algae) organisms.
4.3.2.1.6 Chlorophyll a Determination (Beskenis Draft 2003)
MA DEP has modified the U.S. EPA Fluorometric Method 445.0 to measure chlorophyll
a samples using a Turner Design Fluorometer TD-700. The modified method used does not
require acidification of the sample since background compounds are eliminated with the use of a
filter and blue (mercury) lamp for analysis. Before any samples are run on the fluorometer, it is
calibrated using chlorophyll a standards, and samples are analyzed according to Massachusetts
Division of Watershed Management (MA DWM) methods (Beskenis 2003 Draft). All
chlorophyll a concentrations are reported as mg/m3 and used in conjunction with abundance,
biomass, and percent coverage data to make water quality condition inferences.
4.3.3 Fish Protocols (from Maietta and Decesare 2001)
MA DEP evaluates fish populations using a method based on U.S. EPA RBP V (Plafkin
et al. 1989) to aid in the determination of ALUS classifications in streams. While monitoring
streams for fish, biologists also assess the habitat and characterize the physical and chemical
water quality within the sample reach. A representative 100-m reach is selected and measured
such that the primary physical habitat characteristics of the stream are included within the reach
(i.e., riffle, run and pool habitats, when available). MA DEP selects sample reaches away from
the influences of major tributaries and major bridge and road crossings. Fish are collected using
a pulsed direct current (DC) backpack electrofisher. A crew of at least 3 people begins at a
shallow riffle or other physical barrier at the downstream limit of the stream reach and moves
upstream sweeping from bank to bank until an upstream barrier is reached. If a natural barrier is
not present, block nets are set prior to sampling. All fish caught are added to a bucket or live
well. All wadeable habitats within the reach are sampled with one pass. However a second pass
is required at sites where estimated capture efficiency of observed fish is less than 75%. Fish are
identified to species or subspecies on-site by a qualified/trained fish taxonomist, familiar with
Massachusetts ichthyofauna. Fish are also examined for any deformities, eroded fins, lesions,
and tumors. Any young-of-the-year fishes less than 20 mm in length are not identified or
counted and are released back to the water. A subsample (maximum of 25 specimens) of each
species is measured to the nearest mm in length and weighed to the nearest g. Any unknown
specimens are preserved in 10% formalin solution and transported to the lab for identification by
a second qualified fish taxonomist.
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4.4
Data Management/Quality
All assessment data collected by the MA DEP are entered into the U.S. EPA's Water
Body System (WBS) electronic database. This system aids in the determination of use support
and compiles the data collected throughout the state. The MA DEP also uses a database called
MABEN to analyze macroinvertebrate samples and to determine the condition of the community
at a site using a multimetric approach (Nuzzo 2002). Within the database, taxa richness, HBI (or
FBI), EPT index, ratio of EPT abundances to Chironomidae abundance, ratio of scrapers to
filtering collectors, % contribution of dominant family, and similarity are calculated (Table 4-2).
4.5
Analysis of Biological Data
4.5.1 Macroinvertebrate Data (Nuzzo 2003)
MA DEP uses modifications of the RBPs (II and III) to calculate metrics for the benthic
macroinvertebrate assemblage (Plafkin et al. 1989). The RBP II is designed for family level
taxonomic identification of macroinvertebrates, while the RBP III protocol separates streams
based on generic- and species-level identifications. The metrics are nearly identical between the
two with the exception that the FBI is used for the RBP II (Table 4-2), and the HBI is used for
the RBP III analysis (Table 4-3). Each metric is calculated and then assigned a metric score (6,
3, or 0 for the RBP II; 6, 4, 2, or 0 for the RBP III) based on a comparison with values from
reference sites. Details of metric calculation and scoring are provided in Tables 4-2 and 4-3.
The scores given to each of the individual metrics are then summed to yield a single index value.
Based on the overall index value, the RBP II has the capability to discern between three different
impact categories: Not Impacted, Moderately Impacted, and Severely Impacted. The RBP III
discerns between four different impact categories: Not Impacted, Slightly Impacted, Moderately
Impacted, and Severely Impacted. The overall impact category is determined by comparing the
index score of the test stream to that of the reference site(s), and then determining if the score
lies within the threshold value assigned to the expected level of impairment (Tables 4-2 and 4-3).
Table 4-2. Methods for the calculations of metrics and scoring ranges used in RBP II
determinations of level of biological impact (Plafkin 1989; Nuzzo 2003).
Metric
Taxa Richness (a)
EPT(a)
EPT/Chironomidae
(abundance ratio) (a)
FBI (modified)0^
Method
The total number of distinct taxa in a sample
The number of taxa within the orders of
Ephemeroptera, Plecoptera, and Trichoptera
(Abundance of EPT organisms)/(Abundance of
EPT + Chironomidae)
(Number of individuals in family /') x (Family /'
tolerance value )/(Total number organisms in
sample)
Scoring Ranges
6
>80%
>90%
>75%
>85%
3
40-
80%
70-
90%
25-
75%
50-
85%
0
<40%
<70%
<25%
<50%
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Metric
Scraper/Filtering
Collector Ratio(a)
% Contribution of
Dominant Taxon (G:I
X
T3
>^
-M
1
g
50%
<30%
<0.5
>70%
>64%
3
25-
50%
30-
50%
0.5-
40
30-
70%
35-
64%
0
<25%
>50%
>4.0
<30%
<35%
% Compared to Reference and Impact Category
> 79% Not Impaired
29-72% Moderate Impaired
21% Severe Impaired
a) Value is converted to ratio of test to reference site * 100
b) Value is converted to ratio of reference to test site * 100
c) Actual percent contribution used in scoring, not ratio to reference
d) Uses range of values actually obtained
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Table 4-3. Methods for the calculations of metrics and scoring ranges used in RBP III
determinations of level of biological impact (Plafkin 1989; Nuzzo 2003).
Metric
Taxa Richness(a)
EPT(a)
EPT/ Chironomidae
(abundance ratio)(a)
HBI (modified)(b)
Scraper/Filtering
Collector Ratio(a)
% Contribution of
Dominant Taxon1-0'1
Community Similarity Index
Community
Loss(d)
% Similarity
(d)
% Reference
Affmity(d)
Method
The total number of distinct taxa in a sample
The number of taxa within the orders of
Ephemeroptera, Plecoptera, and Trichoptera
(Abundance of EPT organisms)/(Abundance
of EPT + Chironomidae)
(Number of individuals in taxon /)x(tolerance
value of taxon /)/(Total number of organisms
in sample)
Number of scrapers/filtering organisms.
Number of individuals in most common
taxon/ total number of organisms x 100.
A measure of the dissimilarity between a test
site and a reference site (Plafkin et al. 1989).
Metric values increase as biological
impairment increase. Values have no limits.
CLI = a-c/b
where: a = number of genera in reference
sample, b = number of genera in test sample, c
= number of genera common to both samples
% Similarity = ^ min(a. , bi )
i
Where at is the percentage of taxon / in
sample a and bt is the percentage of taxon / in
sample b.
A test site compared to the reference site for
seven faunal groups (Ephemeroptera,
Trichoptera, Plecoptera, Chironomidae,
Oligochaeta, Coleoptera, and Other).
Calculated similarly to % Similarity but only
percentages for the aggregate groups are used
(Novak and Bode 1992).
Scoring Ranges
6
>80%
>90%
>75%
>85%
>50%
<20%
<0.5
>70%
>64%
4
60-
80%
80-
90%
50-
75%
70-
85%
35-
50%
20-
29%
0.5-
1.5
50-
70%
50-
64%
2
40-
59%
70-
79%
25-
49%
50-
69%
20-
34%
30-
40%
1.6-
4.0
30-
49%
35-
49%
0
<40%
<70%
<25%
<50%
<20%
>40%
>4.0
<30%
<35%
% Compared to Reference Sites
> 83% Not Impaired
54-79% Slight Impaired
21-50% Moderate Impaired
<17% Severe Impaired
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Metric
Method
Scoring Ranges
6
a) Value is converted to ratio of test to reference site * 100
b) Value is converted to ratio of reference to test site * 100
c) Actual percent contribution used in scoring, not ratio to reference
d) Uses range of values actually obtained
4.5.2 Algal Data (Arthur Johnson, Personal Communication)
Algal data are used in aquatic use attainment as an indicator of nutrient enrichment of a
water body. MA DEP has not developed metrics for algae but does use chlorophyll a, percent
cover of green algae, and biomass to determine if the levels of algae are high enough to disrupt
the biological value of the stream. Aquatic life support is considered fully supporting if no algal
blooms are detected, while persistent algal blooms may represent impairment of ALU support.
4.5.3 Fish Data
MA DEP uses a modification of the RBP V (Plafkin et al. 1989) to assess some aspects of
the fish assemblage in streams. A suite of measurements is used to assess the overall structure
and condition of the fish assemblage. These calculations require the taxonomic identification of
fish to the species level and include: number offish species, number of fluvial
specialists/dependents, number of intolerant species and number of salmonid species. Although
MA DEP does not use an IBI, they do use the fish assemblage data with macroinvertebrate
metrics qualitatively to help determine biological attainment. Furthermore, a detailed habitat
assessment (Barbour et al. 1999) is made to accompany the data.
4.5.4 Summary: Determining ALU Support
The MA DEP uses a "weight of evidence" approach to assign ALU designations and
attainment status for monitored streams. Data are collected from the biological (biotic and
habitat), toxicological and chemical components and referenced against criteria (narrative or
numerical) to make a final decision on the ALUS (Table 4-4).
The biological component used to make the decision takes into consideration the
outcome of the macroinvertebrate index (i.e., impairment designation: Tables 4-2, 4-3 and 4-4),
the structure and condition of the fish assemblage, the habitat and flow regime of the stream, and
algal presence as measured by chlorophyll a, biomass, and percent coverage. Once the available
evidence is analyzed and weighed, a site is placed into an attainment category (e.g., Supported or
Impaired) for the water quality class to which it was originally assigned (e.g., A, B or C). These
assignments are then outlined in the 305(b) report and any streams not in attainment are placed
on the Massachusetts 303(d) list.
4-15
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Table 4-4. Biological, toxicological, and chemical parameters that are used collectively to
determine ALUS. Attainment is assigned based on a "weight of evidence" evaluation. (MA
DEP 2003) (Numerical criteria for dissolved oxygen, pH, and temperature can be found in
314 CMR 4.00 (MA DEP 2000). MA DEP uses the recommended limits published by EPA
pursuant to Section 304(a) of the Federal Act for Toxic Pollutant Criteria).
Variable
Support - Data available clearly
indicates support or minor
modification of the biological
community. Excursions from
chemical criteria not frequent or
prolonged and may be tolerated if the
biosurvey results demonstrate
support.
Impaired - There are frequent or severe
violations of chemical criteria, presence
of acute toxicity, or a moderate or severe
modification of the biological
community.
BIOLOGY
Macroinvertebrates:
Rapid Bioassessment
Protocol (RBP) IIP
Fish Assemblage
Habitat and Flow
Eelgrass Bed Habitat
Macrophytes
Plankton/
Periphyton
Non/Slightly impacted
Best Professional Judgment (BPJ)
BPJ
No/minimal loss, BPJ
BPJ
No/infrequent algal blooms
Moderately or Severely Impacted
BPJ
Dewatered streambed due to artificial
regulation or channel alteration, BPJ
Moderate/severe loss, BPJ
Exotic species present, BPJ
Frequent and/or prolonged algal blooms
TOXICITY TESTS**
Water
Column/Ambient
Sediment
>75% survival either 48 hr or 7-day
exposure
>75% survival
<75% survival either 48 hr or 7-day
exposure
<75% survival
CHEMISTRY- WATER* *
Dissolved
oxygen/percent
saturation (MA DEP
1996, U.S. EPA 1997)
pH (MA DEP 1996,
U.S. EPA 1999)
Temperature (MA
DEP 1996, U.S. EPA
1997)
Toxic Pollutants (MA
DEP 1996, U.S. EPA
1999)
Ammonia-N (MA
DEP 1996, U.S. EPA
1999)
Chlorine (MA DEP
1996, U.S. EPA 1999)
Infrequent excursion from criteria,
BPJ (minimum of three samples
representing critical period)
Infrequent excursion from criteria
Infrequent excursion from criteria1
Infrequent excursion from criteria
Ammonia is pH and temperature
dependent2
0.01 1 mg/L (freshwater) or 0.0075
mg/L (saltwater) total residual
chlorine (TRC)3
Frequent and/or prolonged excursion
from criteria [river and shallow lakes:
exceedances >10% of measurements;
deep lakes (with hypolimnion):
exceedances in the hypolimnetic area
>10% of the surface area].
Criteria exceeded >10% of
measurements.
Criteria exceeded >10% of
measurements.
Frequent and/or prolonged excursion
from criteria (exceeded >10% of
measurements).
4-16
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Variable
Support - Data available clearly
indicates support or minor
modification of the biological
community. Excursions from
chemical criteria not frequent or
prolonged and may be tolerated if the
biosurvey results demonstrate
support.
Impaired - There are frequent or severe
violations of chemical criteria, presence
of acute toxicity, or a moderate or severe
modification of the biological
community.
CHEMISTRY-SEDIMENT* *
Toxic Pollutants
(Persaud et al. 1993)
Concentrations < Low Effect Level
(L-EL), BPJ
Concentrations > Severe Effect
EL)4, BPJ
Level (S-
CHEMISTRY-TISSUE
PCB - whole fish
(Coles 1998)
DDT (Environment
Canada 1999)
PCB in aquatic tissue
(Environment Canada
1999)
<500 ng/kg wet weight
<14.0 ng/kg wet weight
<0.79 ng TEQ/kg wet weight
BPJ
BPJ
BPJ
*RBP II analysis may be considered for assessment decision on a case-by-case basis.
**For identification of impairment, one or more of the following variables may be used to
identify possible causes/sources of impairment: NPDES facility compliance with whole effluent
toxicity test and other limits, turbidity and suspended solids data, nutrient (nitrogen and
phosphorus) data for water column/sediments. Maximum daily mean T in a month (minimum
six measurements evenly distributed over 24-hours) less than criterion. 2 Saltwater is temperature
dependent only.3 The minimum quantification level for TRC is 0.05 mg/L. 4For the purpose of
this report, the S-EL for total polychlorinated biphenyl compounds (PCB) in sediment (which
varies with Total Organic Carbon (TOC) content) with 1% TOC is 5.3 ppm while a sediment
sample with 10% TOC is 53 ppm.
4.6
Literature Cited
(APHA) American Public Health Association. 1992. Standard Methods for the Examination of
Water and Wastewater, 18
D.C.
th
edition. American Public Health Association, Washington,
Barbour, M, J. Gerritsen, B.D. Snyder, and J.B. Stribling. 1999. Rapid Bioassessment Protocols
for Use in Streams and Wadeable Rivers: Periphyton, Benthic Macroinvertebrates and
Fish, Second Edition. EPA 841-B-99-002. U.S. Environmental Protection Agency,
Office of Water, Washington, D.C.
http://www.epa.gov/owow/monitoring/rbp/wp61 pdf/rbp. pdf
Beskenis, J. 2002. Standard Operating Procedures, Benthic Algae: Micro and Macro
Identifications and Biomass Determinations. CN: 060.0. Department of Environmental
Protection, Division of Watershed Management, Worcester, MA.
4-17
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Beskenis, J. 2003 (Draft). Standard Operating Procedures. Extracted Chlorophyll a (SM -
10200H) (USEPA Flourometric Method 445 and 445 with the Welschmeyer
modification). CN 0003.0. Department of Environmental Protection, Division of
Watershed Management, Worcester, MA.
Biggs, B.J.F. 1996. Patterns of benthic algae in streams. Pages 31-35 in R.J. Stevenson, M.
Bothwell, and R.L. Lowe (editors). Algal Ecology: Freshwater Benthic Ecosystems.
Academic Press, San Diego, CA.
Bode, R.W. 1990. Chironomidae. Pages 225-267 in B.L.Peckarsky, P.R. Fraissinet, M.A.
Penton, and DJ. Conklin, Jr. (editors). Freshwater Macroinvertebrates of Northeastern
North America. Comstock Publishing Association, Ithaca, NY.
Coffman, W.P. and L.C. Ferrington. 1996. Chironomidae. Pages 635-754 in R.W. Merritt and
K.W. Cummins (editors). An Introduction to the Aquatic Insects of North America.
Kendall/Hunt Publishing Company, Dubuque, IA.
Coles, J.C. 1998. Organochlorine Compounds in Fish Tissue from Connecticut, Housatonic and
Thames River Basins Study Unit, 1992-1994. National Water-Quality Assessment
Program, U.S. Department of the Interior, U.S. Geological Survey, Marlbourough, MA.
Courtemanch, D.L. 1984. A closing artificial substrate device for sampling benthic
macroinvertebrates in deep rivers. Freshwater Invertebrate Biology 3(3): 143-146.
Environment Canada. 1999. Canadian Environmenatal Quality Guidelines. Environment
Canada.
Griffith, G.E., J.M. Omernik, S.M. Pierson, and C.W. Kiilsgaard. 1994. Massachusetts
Ecological Regions Project. EPA/600/A-94/111. U.S. Environmental Protection
Agency, Corvallis, OR.
Jokinen, E.H. 1983. The Freshwater Snails of Connecticut. State Geological and Natural
History Survey of Connecticut, Bulletin 109.
Kathman, R.D. and R.O. Brinkhurst. 1998. Guide to the Freshwater Oligochaetes of North
America. Aquatic Resources Center, College Grove, TN.
Klemm, DJ. (editor). 1985. A Guide to the Freshwater Annelida (Polychaeta, naidid and
tubificid Oligochaeta, and Hirudinea) of North America. Kendall/Hunt Publishing
Company, Dubuque, IA.
Klemm, D.J., P.A. Lewis, F. Fulk, and J.M. Lazorchak. 1990. Macroinvertebrate Field and
Laboratory Methods for Evaluating the Biological Integrity of Surface Waters.
EPA/600/4-90/030. U.S. Environmental Protection Agency, Cincinnati, OH.
4-18
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Maietta, B. and G. Decesare. 2001. Standard Operating Procedures: Fish Collection Procedures
for Evaluation of Resident Fish Populations. CN: 75.0. Department of Environmental
Protection, Division of Watershed Management, Worcester, MA.
(MA DEP) Massachusetts Department of Environmental Protection. 1996. Massachusetts
Surface Water Quality Standards. Massachusetts Department of Environmental
Protection, Division of Pollution Control, Technical Services Branch, Westbourough,
MA.
MA DEP. 2000. 314 CMR 4.00, Massachusetts Surface Water Quality Standards.
Massachusetts Department of Environmental Protection, Division of Water Pollution
Control, http://www.mass.gov/dep/bwp/iww/files/314cmr4.htm
MA DEP. 2003. Massachusetts Year 2002 Integrated List of Waters: Part 1, Context and
Rational for Assessing and Reporting the Quality of Massachusetts Surface Waters.
CN: 125.1. Massachusetts Department of Environmental Protection, Division of
Watershed Management, http://www.mass.gov/dep/brp/wm/files/2002-il2.pdf
Merritt, R.W., and K.W. Cummins (editors). 1996. An Introduction to the Aquatic Insects of
North America. Kendall/Hunt Publishing Company, Dubuque, IA.
Novak, M. A., and R. W. Bode. 1992. Percent model affinity: a new measure of
macroinvertebrate community composition. Journal of the North American
Benthological Society 11:80-85.
Nuzzo, R.M. (editor). 2002. Benthic macroinvertebrate database descriptions and procedures
(rev. 1.1). Massachusetts Department of Environmental Protection, Division of
Environmental Management, Worcester, MA.
Nuzzo, R.M. 2003. Standard Operating Procedures. Water Quality Monitoring in Streams
Using Aquatic Macroinvertebrates. Massachusetts Department of Environmental
Protection, Division of Watershed Management, Worcester, MA.
Peckarsky, B.L., P.R. Fraissinet, M.A. Penton, and DJ. Conklin, Jr. 1990. Freshwater
Macroinvertebrates of Northeastern North America. Comstock Publishing Association,
Ithaca, NY.
Persaud, D., R. Jaaguamagi, and A. Hayton. 1993. Guidelines for the Protection and
Management of Aquatic Sediment Quality in Ontario. Water Resources Branch, Ontario
Ministry of the Environment, Queen's Printer for Ontario, Canada.
Plafkin, J.L., M.T. Barbour, K.D. Porter, S.K. Gross, and R.M. Hughes. 1989. Rapid
Bioassessment Protocols for Use in Streams and Rivers: Benthic Macroinvertebrates and
Fish. EPA/444/4-89-001. U.S. EPA, Washington D.C.
4-19
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Smith, D.G. 1986. Keys to the Freshwater Macroinvertebrates of Massachusetts (No. 1):
Mollusca Pelecypoda (clams, mussels). Massachusetts Division of Water Pollution
Control, Westbourough, MA.
Smith, D.G. 1987. Keys to the Freshwater Macroinvertebrates of Massachusetts (No. 2):
Mollusca Mesogastropoda (operculate snails). Massachusetts Division of Water
Pollution Control, Westborough, MA.
Smith, D.G. 1988. Keys to the Freshwater Macroinvertebrates of Massachusetts (No. 3):
Crustacea Malacostraca (crayfish, isopods, amphipods). Massachusetts Division of Water
Pollution Control, Wesborough, MA.
U.S. EPA. 1997. Guidelines for Preparation of the Comprehensive State Water Quality
Assessments (305(b) Reports) and Electronic Updates: Supplement. U.S. Environmental
Protection Agency, Office of Water, Office of Wetlands, Oceans and Watersheds,
Assessment and Watershed Protection Division, Washington, D.C.
U.S. EPA. 19 November 1999. Federal Register Document. [Online]. Environmental Protection
Agency. http://www.epa.gov/fedrgstr/EPA-WATER/1998/December/Dav-
10/w3 0272. htm
Wiederholm, T. (editor). 1983. Chironomidae of the Holarctic Region: Keys and Diagnoses
(Part 1. Larvae). Entomologica Scandinavica, Supplement No. 19.
Wiederholm, T. (editor). 1986. Chironomidae of the Holarctic Region: Keys and Diagnoses
(Part 2. Pupae). Entomologica Scandinavica, Supplement No. 28.
4.6.1 Resources
Massachusetts Department of Environmental Protection, Bureau of Resource Protection Web
Page: http://www.state.ma.us/dep/brp/
U.S. EPA. Biological Indicators of Watershed Health, Massachusetts Webpage:
http://www.epa.gov/bioindicators/html/state/ma-bio.html
U.S. EPA. 2002. Summary of Biological Assessment Programs and Biocriteria Development or
States, Tribes, Territories, and Interstate Commissions: Streams and Wadeable Rivers.
EPA-822-R-02-048. U.S. Environmental Protection Agency.
4-20
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5 NEW HAMPSHIRE
This document was prepared using documents written by the State of
New Hampshire. Any questions concerning bioassessment methods
should be directed to:
Dave Neils, Biomonitoring Program Coordinator
New Hampshire Department of Environmental Services (NH DES)
Concord, NH 03302-0095
Phone: (603) 271-8865; Fax (603) 271-7894
Email: dneils@des.state.nh.us
5.1 Introduction
The State of New Hampshire Water Quality Standards (NH WQS), as defined by the NH
DES, recognizes six designated uses in surface freshwaters: aquatic life, drinking water supply,
fish consumption, primary contact recreation, secondary contact recreation, and wildlife. The
narrative standards for water use classifications are stated as: "(a) State surface waters shall be
divided into class A and class B, pursuant to RSA 485-A:8,1, II and III. Each class shall identify
the most sensitive use which it is intended to protect; (b) All surface waters shall be restored to
meet the water quality criteria for their designated classification including existing and
designated uses, and to maintain the chemical, physical, and biological integrity of surface
waters; (c) All surface waters shall provide, wherever attainable, for the protection and
propagation offish, shellfish and wildlife, and for recreation in and on the surface waters; (d)
Unless the flows are caused by naturally occurring conditions, surface water quantity shall be
maintained at levels adequate to protect existing and designated uses" (NH DES 1999).
Furthermore, NH DES standards define biological and aquatic community integrity such
that "a) The surface waters shall support and maintain a balanced, integrated, and adaptive
community of organisms having a species composition, diversity, and functional organization
comparable to that of similar natural habitats of a region" and where "b) Differences from
naturally occurring conditions shall be limited to non-detrimental differences in community
structure and function" (NH DES 1999). For rivers and streams and associated impoundments
(4th order or less), ALUS is determined based on ten indicators, with the macroinvertebrate
assemblage as the core indicator of biological condition (NH DES 2004a). The other indicators
are: dissolved oxygen, pH, habitat, water quality criteria for toxic substances, toxicity tests of
ambient water, sediment quality, exotic macrophytes, flow, and benthic deposits. In all other
surface waters, ALUS is determined based on biological assemblage data or a minimum of DO
and pH. If these constituents are within allowable standards or if there is documentation from a
trained biologist that there is no other obvious impairment to the biological community, then the
water body can be listed as fully supporting.
5-1
-------
Table 5-1. NH DES water quality classes and the defined designated uses for each class.
Dissolved oxygen exceedance values for aquatic life criteria are also listed (NH DES 1999 ).
Classification
Designated Uses
Exceedances of the water quality criteria
(aquatic life) for dissolved oxygen
(Applies to any depth in free flowing rivers)
Class A
Generally of the
highest quality and
considered
potentially usable
for water supply
after adequate
treatment.
Discharge of
sewage or wastes
prohibited to waters
of this
classification.
Daily Average Measurement: < 75% saturation;
Instantaneous Measurement: <6 mg/1
In cold water naturally producing fisheries
Daily Average Measurement: From October 1 to
May 14, a 7 day mean DO based on the daily
average of < 9.5 mg/L;
Instantaneous Measurement: From October 1 to
May 14 DO < 8 mg/L
Class B
Of the second
highest quality,
considered
acceptable for
fishing, swimming
and other
recreational
purposes, and, after
adequate treatment,
for use as water
supplies.
Daily Average Measurement:<75% saturation;
Instantaneous Measurement: < 5mg/l
In cold water naturally producing fisheries
Daily Average Measurement: From October 1 to
May 14, a 7 day mean DO based on the daily
average of < 9.5 mg/L;
Instantaneous Measurement: From October 1 to
May 14 DO < 8 mg/L
The NH DES uses a weight of evidence approach when taking in consideration
biological, RBP habitat, in situ chemistry, physical (e.g., landuse coverages and point sources)
and toxicological data to support narrative criteria that determine ALUS (NH DES 2004a).
Biological and physical habitat data are given the highest weight as "they are a direct
measurement of the aquatic life and detect the cumulative impact on the aquatic community
including new or previously undetected stressors over time" (NH DES 2004a). NH DES collects
fish and macroinvertebrate data in their biomonitoring program to assess the condition of streams
for 305(b) reporting and 303(d) listing. A benthic macroinvertebrate multimetric index has been
developed recently, and a fish index is currently under development through the modification of
the Vermont Cold Water Index of Biotic Integrity (CWIBI) and Mixed Water Index of Biotic
Integrity (MWIBI) (see chapter 7 in this document, as well as VT DEC 2004). All other data
collected (chemical, physical habitat and toxicological) provide complementary information and
assist the biologists in determining ALUS.
5-2
-------
5.2 Key Elements of the Biological Assessment Approach
5.2.1 Index Period and/or Temporal Conditions
The NH DES biomonitoring program collects both macroinvertebrates and fish during
the mid-summer to early fall index period. This period represents the lowest flows and most
stressful hydrologic conditions for instream biota. During this period, the larvae of the
macroinvertebrate assemblages are easily identifiable because the late instars are larger.
Furthermore, the fish assemblage is more stable and resident species are more likely to remain in
a localized area when flow conditions are not fluctuating dramatically. The sampling index
period is consistent from year to year in order to accurately compare monitoring data through
time.
5.2.2 Monitoring Program Survey Approach
When developing the biocriteria program, an effort was made by NH DES to initially
sample least impacted sites from a variety of habitats. Since then the program has collected
samples from moderate and high impact sites in order to develop a multimetric index that best
represents a response of the biological community to human disturbance. Sites have also been
chosen for macroinvertebrate and fish monitoring using a targeted approach for 305(b) reporting
and for identifying sites that cover geographical data gaps. Approximately 25-30 sites are
sampled statewide per year.
5.2.3 Natural Classification of Water Bodies
New Hampshire consists of five major river basins: Androscoggin, Connecticut, Saco,
Merrimack, and Piscataqua (Figure 5-1). For macroinvertebrate biomonitoring purposes, the
state is separated into bioregions defined by distinct biological community types. The
boundaries for the "Northern" and "Southern" bioregions represent similar Ecological Drainage
Units as defined by The Nature Conservancy (Figure 5-1). For fish, streams are divided into two
categories: cold and warm water.
5.2.4 Indicator Assemblages
Currently (for the 2004 305 (b) report), NH DES is using the macroinvertebrate
assemblage to determine ALUS. A Benthic Index of Biotic Integrity (B-IBI) is used to assess
the macroinvertebrate assemblages (with separate biocriteria for northern and southern
bioregions). A fish Index of Biotic Integrity (IBI) is currently being developed to assess the fish
assemblages of both cold and mixed waters. NH DES is basing the fish IBI on Vermont's
CWIBI and MWIBI because of the presumed similarity in fish assemblage composition,
structure, and function.
5.2.5 Reference Condition
NH DES defines reference condition as the least disturbed sites. Reference sites were
chosen based on a state-wide scoring system that takes into account both local and watershed
5-3
-------
scale variables using GIS coverages of land use and point sources, as well as an immediate in-
stream measure of human influence using the U.S. EPA's RBP habitat assessment (Barbour et al.
1999). On a watershed scale, land use was portioned into percentages developed, undeveloped
and agricultural lands categories using the New Hampshire Land Cover (NHLC) dataset.
Androscnggin
River Basin
Northern Bioregion
Connecticut
River Basin
Menimack
River Basin
Southern Bioregion
Figure 5-1. Major New Hampshire basins and the northern and southern bioregion
boundaries used for macroinvertebrate sampling (indicated by the red line).
Percent water impounded was also calculated at the watershed scale. Local scale variables were
estimated for a 300-ft buffer on either side of all hydrologic features within the watershed and
within a 1-mile radius of the site. Densities of Ground Water Hazard Inventory (GWHI) sites,
Resource Conservation and Recovery Act (RCRA) sites, junkyards, dams, water withdrawals,
National Pollutant Discharge Elimination System (NPDES) sites, and roads were calculated at
this local scale for each site. Each variable was scored on a 0-3 scale, with 0 representing
5-4
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minimal disturbance for that variable and 3 representing maximum disturbance, based upon
overall distributions across all sites sampled between 1997 and 2001. Scores were summed for
both the local and watershed scale disturbance scores, as well as across all variables to yield a
total disturbance score (the human disturbance gradient (HDG) score). The total score was
divided into four disturbance levels with equal scoring ranges: 0-4 (Best), 5-9 (Good), 10-14
(Fair), and >14 (Worst). The entire range observed across all sites was 0 to 19. Those sites
scoring in the best range were identified as the least disturbed sites. This HDG resulted in the
identification of 46 reference condition sites within New Hampshire, 26 in the northern bioregion
and 20 in the southern region.
5.3 Field and Laboratory Protocols
5.3.1 Macroinvertebrates Protocols
5.3.1.1 Field Methods (NH DES 2004b)
NH DES uses rock baskets to collect macroinvertebrates for B-IBI calculations. Rock
baskets consist of an 11-in cylindrical plastic coated wire barbecue basket with 1-in2 mesh
openings. The bottom of the basket has a hinged opening. The basket is filled with regionally
indigenous bank run gravel ranging in diameter from 1.5-3.0 in. The hinge is then secured with a
plastic tie wrap. Baskets are placed in groups of three replicates attached to a Va-in steel rod
anchor. Baskets are submerged to depths of greater than 5 in. Each of the attached baskets is
arranged so that the bottom is facing downstream.
The baskets are left in the stream for eight weeks to allow for colonization. At removal,
the baskets are approached from downstream and a 3-gal bucket containing a 600-jim sieve
bottom is placed downstream of the rock baskets. The plastic cable ties that secure the three
baskets to the steel rod are cut and the baskets are quickly placed into the bucket. Any extra
debris or algae clinging to the outside of the rock baskets are removed and discarded. The
bucket containing the baskets is then transported to the stream side for sample retrieval. Using a
knife, the plastic tie wraps are cut and the rocks are emptied into the sieve bucket. Then, the
empty rock basket is added to a 5-gal bucket containing 3-4 gal of water. The basket is scraped
free of organisms using a soft bristle brush. The 5-gal bucket is then emptied into the sieve
bucket. Next, 2-3 gal of water are added to the 5-gal bucket and the sieve bucket is nested
inside. Using a soft bristle brush, the rocks inside the sieve bucket are scrubbed free of attached
organisms, and each scrubbed rock is then returned to the rock basket. After all organisms and
detritus have been removed from the rocks, the sieve bucket is lifted from the 5-gallon bucket,
capturing the targeted sample in the sieve. The contents of the sieve bucket are then placed into
a one-quart, wide-mouthed jar and preserved with a solution of 1/3 water and 2/3 ethanol. This
process is repeated for each of the replicate baskets. The scraped rocks from the baskets are
allowed to dry before baskets are reused.
5.3.1.2 Laboratory Methods (NH DES 2004b)
NH DES processes macroinvertebrate samples collected in rock baskets using the U.S.
EPA Caton Method (Caton 1991). A tray is fitted with a gridded screen with 16 uniform
squares. Enough water is added to the tray so that the entire sample can be dispersed evenly over
5-5
-------
the screen. The screen is then removed so that the organisms are settled on the screen. Random
numbers are then selected to choose the grids from which to sort organisms. A metal square is
used to delineate a grid square and the organisms are removed using a scoop. Those organisms
that occupy more than one grid are counted in the grid that contains the head, unless the head is
missing. Then, the grid with the largest portion of the organism is counted. At least 25% of the
squares are counted yielding a minimum sample of 100 organisms. In the event that the 100-
organism target is not achieved, the entire sample is sorted. After sorting, benthic organisms are
identified to the lowest possible taxonomic level. For data analysis, however, all Chironomidae
were aggregated to the family level.
5.3.2 Fish Protocol (NH DES 2004b)
NH DES follows U.S. EPA's RBP V (Barbour et al. 1999) to collect fish from wadeable
streams using backpack electrofishing units. For wadeable streams, a minimum reach length of
150 m is selected in close proximity to the location where the macroinvertebrate rock baskets
were placed. NH DES has established a minimum 150-m reach as "the reasonable limit to
prevent oversampling, while optimizing efficiency and representation of the resident species"
(NH DES 2004b). A field sampling crew comprised of one shocker, at least two netters, and one
person to carry an aerated bucket, begins at the most downstream portion of the reach and moves
upstream. The crew collects a representative fish sample along the available instream habitat
using a single-pass method. All fish identifications are made streamside. The identities, number
offish and external anomalies are recorded, and the fish are then released. For important cold
water game fish, young-of-the-year (YOY) are recorded in order to document natural spawning
activity. Fish that cannot be easily identified are taken back to the laboratory for identification
by a trained biologist or using the fishes of New Hampshire key (Scarola 1973).
5.4 Data Management/Quality
NH DES has adapted the Ecological Data Assessment System (EDAS) to manage the
chemical, physical and biological data collected in New Hampshire (EDAS 1999). EDAS is a
Microsoft Access-based data storage and retrieval warehouse that contains station information,
basic water chemistry data, fish data, habitat assessments, macroinvertebrate data and flow data.
Updated GIS coverages of sample locations and upstream watersheds are also maintained using
ARCVIEW software. The GIS coverages include data pertaining to watershed size, station
elevation, latitude, longitude, stream order, Human Disturbance Gradient (HDG) status,
bioregion, and hydrologic unit code (HUC). The NH DES uses the Assessment Database (ADB)
developed by the U.S. EPA to submit electronic reports. Since 2002, the new Oracle-based
version has been used by NH DES to submit surface water assessments.
5.5 Analysis of Biological Data
5.5.1 Macroinvertebrate Data (Neils and Blocksom 2004)
The following seven metrics comprise the New Hampshire Benthic Index of Biotic
Integrity (B-IBI): Total number of taxa, number of Plecoptera taxa, % Chironomidae, % non-
insects, % clingers, number of tolerant taxa, and % individuals in intolerant taxa. NH DES used
5-6
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the 95th percentile across all sites as a threshold for metrics that decrease in response to
disturbance (i.e., positive metrics) and the 5th percentile across all sites for metrics that increase
in response to disturbance (i.e., negative metrics) (Table 5-2). Scoring for each metric is on a
continuous scale ranging from 0 to 100. Positive metrics are scored using the following
equation:
oh served
SCORE = ooservea *iOO
threshold
Negative metric scores are calculated using the following equation:
SCORE = WO
(max- threshold)
The maximum is either the maximum possible (for percentage metrics) or the maximum
observed in the calibration data set (for all other metrics). The metric scores for each site are
calculated and averaged across all metrics, and the score is compared to the bioregional threshold
value.
Within each bioregion, the distribution of reference sites was used to set the biocriterion
for the New Hampshire B-IBI. NH DES does not regard the reference sites used as truly
unimpaired conditions for the state and has set the thresholds for attainment of ALU standards to
take into consideration the possibility of incomplete information about sites. The 25th percentile
of the reference distribution within each bioregion was used as the biocriterion. In the Northern
Region, the threshold score is 77, and in the Southern Region, the threshold score is 66.4. Sites
with B-IBI scores at or above the threshold in each bioregion are considered to be in attainment
ofALUS.
5.5.2 Fish Data (personal communication, David Neils, NH DES; VT DEC 2004)
Currently, NH DES is modifying the Vermont Cold Water Index of Biotic Integrity
(CWIBI) and the Mixed Water Index of Biotic Integrity (MWIBI) for use in New Hampshire
streams. Detail of the Vermont indices can be found in Chapter 7. Vermont uses the CWIBI in
streams that contain two to four species, and the MWIBI in streams that contain greater than four
species (VT DEC 2004). NH DES used native fish species richness to adapt the method to use
the MWIBI for any stream containing five or more taxa and used the CWIBI for other streams
containing less than five taxa. Best professional judgment was used to ensure a stream was
placed into the correct category. After this initial categorization, sites identified as belonging in
MWIBI category were further refined as cold or warm water fish communities utilizing Eastern
Brook Trout and Slimy Sculpin as indicator species. Site elevations were also used to classify
MWIBI sites into cold and warm fish communities. Within the MWIBI, the differentiation
between cold and warm water fish assemblages was applied in scoring individual metrics.
Although the use of the CWIBI and MWIBI is currently under development, NH DES found the
indices to be promising because the method showed significant differences between reference
and non-reference sites. Further refinement of the CWIBI and MWIBI will be completed in the
near future to calibrate final index scores and establish attainment cutoffs specific to New
5-7
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Hampshire fish communities. Although the CWIBI or MWIBI appeared to work well for many
sites, as in VT, neither index is applicable in slow, winding sites.
Table 5-2. Metrics and scoring for the New Hampshire B-IBI.
Metric
Total taxa
Plecoptera taxa
%
Chironomidae
% Non-insects
% Clingers
% Intolerant
Tolerant taxa
Definition
Number of distinct
macroinvertebrate
taxa.
Number of taxa in
the order Plecoptera
(genus or species
level).
% individuals in the
family
Chironomidae.
% individuals in a
sample that are not
in the class Insecta.
% insects having
fixed retreats or
adaptations for
attachment to
surfaces in flowing
water.
% of individuals
considered to be
sensitive to various
types of pollution.
Taxa richness of
those organisms
considered to be
tolerant to
increased
disturbance.
Expected
Response to
Disturbance
Decrease
Decrease
Increase
Increase
Decrease
Decrease
Increase
Scoring Equation
Total taxal2\.5*\W
Plecoptera tara/4.4*100
(100 - % Chironomidae)/ 100-
0)*100
(100- % Non-insects)! 100-
0)*100
%Clingers/94.6*WO
% Intolerant/7 6. 1*100
(6.2-Tolerant fajear)/(6.2-0)*100
5.6
Summary: Determining ALU Support
NH DES follows the guidelines listed in the Comprehensive Assessment and Listing
Methodology (CALM) to assess streams for ALUS (NH DES 2004a). This document lists the
assessment criteria necessary for making the decision for specific segments (Assessment Units,
AUs) of wadeable streams. Non-support in an ALU segment can be determined based on the B-
IBI score or the failure to meet chemical criteria (e.g., DO or pH). However, Full Support status
cannot be given to a segment without a B-IBI evaluation within that segment. This assessment
must also include data from the most recent calendar year. Other data, such as RBP habitat
5-S
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assessments, fish assessments, benthic deposits, flow, macrophyte composition, sediment and
ambient water toxicity tests are also used to aid in the decision-making process.
5.7 Literature Cited
Barbour, M, J. Gerritsen, B.D. Snyder, and J.B. Stribling. 1999. Rapid Bioassessment Protocols
for Use in Streams and Wadeable Rivers: Periphyton, Benthic Macroinvertebrates and
Fish, Second Edition. EPA 841-B-99-002. U.S. Environmental Protection Agency,
Office of Water, Washington, D.C.
http://www.epa.gov/owow/monitoring/rbp/wp61 pdf/rbp. pdf
Caton, L.W. 1991. Improving subsampling methods for the EPA "Rapid Bioassessment"
benthic protocols. Bulletin of the North American Benthological Society 8(3):317-319.
Ecological Data Assessment System (EDAS). 1999. Tetra Tech, Inc.
Neils, D., and K.A. Blocksom. 2004. Development of the New Hampshire Benthic Index of
Biotic Integrity. New Hampshire Department of Environmental Services, Concord, New
Hampshire.
(NH DES) New Hampshire Department of Environmental Services. 1999. State of New
Hampshire Surface Water Quality Regulations, Chapter 1700.
http://www.des.state.nh.us/wmb/env-wsl700.pdf
NHDES. 2004a. 2004 Section 305(b) and 303(d) Consolidated Assessment and Listing
Methodology. New Hampshire Department of Environmental Services, Concord, NH.
http://www.des.state.nh.us/wmb/swqa/2004/pdf/CALM.pdf
NHDES. 2004b. Biomonitoring Program Protocols. New Hampshire Department of
Environmental Services, Concord, NH.
Plafkin, J.L., M.T. Barbour, K.D. Porter, S.K. Gross, and R.M. Hughes. 1989. Rapid
Bioassessment Protocols for Use in Streams and Rivers: Benthic Macroinvertebrates and
Fish. EPA 440-4-89-001. U.S. Environmental Protection Agency, Office of Water
Regulations and Standards, Washington, D.C.
Scarola, J.F. 1973. Freshwater Fishes of New Hampshire. New Hampshire Fish and Game
Department, Division of Inland and Marine Fisheries.
(VT DEC) Vermont Department of Environmental Conservation. 2004. Biocriteria for Fish and
Macroinvertebrate Assemblages in Vermont Wadeable Streams and Rivers -
Implementation Phase. Vermont Department of Environmental Conservation,
Waterbury, VT. http://www.vtwaterquality.org/bass/htm/bs_biomon.htm
5-9
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5.8 Resources
New Hampshire Department of Environmental Services, Watershed Management Bureau,
Biomonitoring Program Web Page: http://www.des.state.nh.us/wmb/biomonitoring/
U.S. EPA. Biological Indicators of Watershed Health, New Hampshire Webpage:
http://www.epa.gov/bioindicators/html/state/nh-bio.html
U.S. EPA. 2002. Summary of Biological Assessment Programs and Biocriteria Development
for States, Tribes, Territories, and Interstate Commissions: Streams and Wadeable Rivers.
EPA-822-R-02-048. U.S. Environmental Protection Agency.
http://www.epa.gov/bioindicators/html/program summary.html
5-10
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6 RHODE ISLAND
This document was prepared using documents written by the State
of Rhode Island. Any questions concerning bioassessment
methods should be directed to:
Connie Carey, Principal Environmental Scientist
Rhode Island Department of Environmental Management (RI
DEM)
Office of Water Resources
235 Promenade Street
Providence, Rhode Island 02908
Phone: (401) 222-3961 ext. 7239; Fax: (401) 222-3564
Email: ccarey@dem.state.ri.us
6.1 Introduction
Rhode Island WQS define water quality goals for the state's waters by designating uses
and setting criteria necessary to protect those uses. Rhode Island WQS provide for the
protection of the waters from pollutants so that the waters shall, where attainable, be available
for all designated uses (i.e., drinking water supply, shell fish consumption, fish consumption,
swimming, aquatic life) and thus assure protection for the public health, welfare, and the
environment (RI DEM 2000). Rhode Island WQS define ALU as "providing suitable habitat
and water quality for the protection, maintenance, and propagation of a viable community of
aquatic life". In accordance with Section 305(b) of the CWA, states are required to evaluate
water quality of all water body types (i.e., rivers/streams, lakes/ponds, estuarine waters) for
attainment of their designated uses. The water quality criteria and assessment methodology used
for the evaluation is detailed in the Integrated Water Quality Monitoring and Assessment Report.
Any water bodies that are not attaining their designated uses are placed on the 303(d) List of
Impaired Waters and a TMDL must be developed for each exceedance that results in non-
attainment status.
The Rhode Island Department of Environmental Management (RI DEM) has used
biological assessments to supplement physical and chemical water quality monitoring data for
evaluation of ALU attainment in rivers and streams. The Rhode Island WQS currently only
contain narrative biological criteria to utilize in evaluating biological assessment data. RI DEM
implements a reference site approach for evaluation of the macroinvertebrate assemblage, as
described below. The data collected from the bioassessment program is compared to
corresponding reference sites and scored relative to conditions observed at the reference station.
Scoring information from multiple years is used to determine a site's ALU attainment status for
305(b) reporting and 303(d) listing purposes.
6-1
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6.2 Key Elements of the Biological Assessment Approach
6.2.1 Index Period and/or Temporal Conditions
The RI DEM conducts its biological monitoring program during the summer and fall
seasons. Sites are sampled annually during this index period and any data collected and
evaluated during drought conditions are noted. A long-term database allows for current and
historical comparisons of data using these seasons.
6.2.2 Monitoring Program Survey Approach
Currently, RI DEM samples 45 fixed-river stations located throughout the state. The
sampling sites are located within one of two subecoregions in Rhode Island: the New England
Coastal Plains and Hills, or the Narragansett/Bristol Lowland. The stations were originally
located on rivers/streams that were considered unassessed for ALU. They generally consist of
smaller, wadeable rivers that do not have point-source discharges located on them, and range
from first order to fifth order in size.
6.2.3 Natural Classification of Water Bodies
Rhode Island has two Level IV Omernik subecoregions represented within the
Northeastern Coastal Zone ecoregion: the New England Coastal Plains and Hills subecoregion
and the Narragansett/ Bristol Lowland subecoregion. Each subecoregion is represented by a
reference site that is used to compare to test sites within the subecoregion.
6.2.4 Indicator Assemblages
RI DEM currently uses only the macroinvertebrate assemblage as a biological indicator.
Macroinvertebrates are typically identified to the lowest practical taxonomic level. Rhode Island
follows the U.S. EPA's RBP for evaluation of benthic macroinvertebrate communities (Plafkin et
al. 1989).
6.2.5 Reference Condition
The RI DEM currently uses two reference sites in their bioassessment program, one
representing each of the two Level IV Omernik subecoregions present in the state (Figure 6-1).
One reference site, the Wood River, is located in the New England Coastal Plains and Hills
subecoregion, while Adamsville Brook serves as the reference site for the Narragansett/Bristol
Lowland subecoregion. Currently, no specific reference criteria exist, but each reference site
represents minimally disturbed, high quality, historically natural sites within its subecoregion.
The Wood River site is located on a fourth order, minimally disturbed portion of the river within
the Pawcatuck River Basin. This reference site is located almost completely within the
boundaries of a state park, and, therefore, is not expected to undergo the degradation that would
remove it from the reference category. The Adamsville Brook reference station is located on a
second order portion of the brook in the Cape Cod Basin. This area consists of very low density
residential and 82% rural land use.
6-2
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New England Coastal
Plains and Hills
subeco region
Mai ran (janselt Bristol
Lowland subecoregion
Figure 6-1. Level IV Omernik subecoregions and reference streams used in RI DEM's
biological monitoring program.
6.3 Field and Laboratory Protocols
6.3.1 Macroinvertebrate Protocols
6.3.1.1 Field Methods (RI DEM 2002a, RI DEM 2003)
RI DEM uses the U.S. EPA's RBP III Single Habitat Approach (Barbour et al. 1999) to
assess the macroinvertebrate assemblage of wadeable streams. This method has been in use
since 1991 at the 45 fixed-station riffle stream sites. A physical evaluation is conducted at each
of the sites that includes: surrounding land use; subsystem classification; documentation of dams,
erosion and non-point source pollution; width, depth and flow measurements; inorganic and
organic substrate types; and presence of odors, oils and deposits. Along with a physical
evaluation, water quality measurements are taken (DO, pH, specific conductance, turbidity, and
water temperature), and a habitat assessment is conducted. The habitat assessment and
biological data are scored and compared against the reference station's data. The information is
6-3
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then integrated with water quality, physical, rainfall, and historical trends data, to determine the
ALU attainment status relative to the reference site from the same subecoregion.
RI DEM uses the RBP III Single Habitat Approach to collect representative samples of
the stream reach. A 100-m reach is chosen for sampling, assuring that the reach is located at
least 50 m upstream of bridges or road crossings to minimize their effect on stream velocity,
depth and overall habitat quality. An effort is also made to not sample immediately downstream
of stream confluences. The reach is approached from the downstream to upstream direction
collecting a composite sample of macroinvertebrate kick samples from riffles (multiple if
present) with a dominant cobble substrate. A 0.3-m wide D-Frame net with 500-|im mesh is
placed into the bottom of the riffle, and the substrate is disturbed for three minutes. Larger
substrate particles and debris are rubbed to remove attached organisms. Collected material is
rinsed using stream water before being transferred to a sample container. If multiple riffles are
sampled, all contents of the net from each riffle are placed into a single jar. The sample is
preserved with 70% ethanol.
6.3.1.2 Laboratory Methods (RI DEM 2002a)
Samples preserved in 70% ethanol are rinsed thoroughly with tap water through a 500-|im
mesh sieve to remove fine sediment and preservative. Any large organic material is discarded
and the sample is then distributed evenly in an 18-in x 13-in x 1-in gridded tray with eight
equally-sized squares. The entire sample is scanned, and any rare or large organisms are
removed, identified and used only to report as supplemental data. Then, one section is randomly
selected and all of the material in that section is removed using a 6-cm flat scoop and transferred
to a separate container. Any overhanging debris is cut using scissors. Macroinvertebrates are
sorted under a dissecting microscope on a clean Petri dish and placed into one of the three
following groups in glass vials containing 70% ethanol for identification: 1) Oligochaetes and
Chironomids, 2) Crustaceans and Mollusks, and 3) other organisms. Additional, randomly
selected sections are completely sorted until 100 organisms are sorted or until the entire sample
has been inspected for macroinvertebrates. Random quality checks are performed to assure that
there is less than a 10% discrepancy between the sorter and the quality assurance check.
After sorting, macroinvertebrates are identified to the lowest practical taxonomic level.
Vials containing chironomids and oligochaetes are sent to a sub-contractor, ARC, where they are
mounted on labeled slides using an appropriate medium (e.g., Eupcral, CMC-9) and identified.
All organisms are identified using a compound microscope or a dissecting microscope (up to
45X magnification), a fiber optic lamp, standard dissecting tools, and taxonomic keys.
6.4 Data Management/Quality
RI DEM has recently developed a Microsoft Access database to maintain the data
collected by the biological monitoring program. Portions of the historic biological data have
been entered into the database. Because the Rhode Island Office of Water Resources, where the
biological monitoring program resides, has been without staff for over a year, the more recent
data are maintained only in hard copy format. The Access database has been developed to
calculate RBP metrics for macroinvertebrate data and will assist in the evaluation of developing
a biological condition matrix for the state.
6-4
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6.5
Analysis of Biological Data
6.5.1 Macroinvertebrate Data (RI DEM 2002a, RI DEM 2002b)
RI DEM uses the RBP III (Plafkin et al. 1989) metric-based method to give a
bioassessment score relative to the reference site. Eight RBP metrics and three supplemental
metrics (i.e., EPT abundance, Shannon Weaver Diversity Index, and % Hydropsychidae of total
Trichoptera) are calculated (Table 6-1). Each metric is given a score (6, 4, 2, or 0), primarily
based on the percentage of the reference site value observed at a site. The overall score is the
sum of metric scores. Then, the overall score is compared to the reference site score by
calculating the percentage of the reference score achieved. Thresholds of percentages relative to
the reference site score places a stream into one of four bioassessment categories: Non-impaired,
Slightly Impaired, Moderately Impaired, or Severely Impaired (Table 6-2). Although Table 6-2
lists the threshold percentages for assignment to a particular bioassessment category, the actual
threshold may be adjusted slightly based on best professional judgment of the assessor, as well as
analysis and comparison to historical data and trends.
Table 6-1. Metrics used by the Rhode Island Biomonitoring program and the methods for
the calculation of metrics and their scoring ranges based on the RBP III (Plafkin et al.
1989, RI DEM 2002a, RI DEM 2002b).
Metric
Total Taxa Richness
(a)
EPT Taxa Richness
(a)
EPT Abundance
Hilsenhoff Biotic
Index (b)
Shannon Weaver
Diversity Index
% Contribution of
Dominant Taxon ( )
EPT/Chironomidae
(abundance ratio)(a)
Method
The total number of distinct taxa
in the sample
The number of taxa within the
orders of Ephemeroptera,
Plecoptera, and Trichoptera.
All Ephemeroptera, Plecoptera,
and Trichoptera individuals are
added together.
(Number of individuals in taxon
/')x(tolerance value of taxon
/')/(Sum of individuals with
tolerance values in sample)
Number in each species is counted
and index is calculated as:
£ (p; Iog2 pO
Where p; = the proportion of
individuals in the ith species.
((a/b)x 100)
Where: a = the number of
individuals in the dominant taxon,
b = the total number of individuals
recorded at the stream segment.
(No. of EPT individuals)/(No.
Chironomidae individuals)
Scoring Ranges
6
>80%
>90%
4
60-
80%
80-
90%
2
40-
60%
70-
80%
0
<40%
<70%
Only used as supplementary
data, not used in the RBP
approach.
>85%
70-
85%
50-
70%
<50%
Used as a measure of aquatic
environmental health, but not
used for RBP III.
<20%
>75%
20-
30%
50-
75%
30-
40%
25-
50%
>40%
<25%
6-5
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Metric
Method
Scoring Ranges
0
Percent
Hydropsychidae of
Total Trichoptera
(No. Hydropsychidae
individuals)/(No. all other
Trichoptera) x 100
Used only as additional potential
tolerance/intolerance measure
metric (RI DEM 2003; modified
from Barbour et al. 1999)
Ratio of Shredders to
Total Number of
Macroinvertebrates (a)
(No. shredder individuals)/(Total
no. of other individuals) (RI DEM
2003)
>50
35-
50%
20-
35%
<20%
Ratio of Scrapers to
Filterers(a)
(No. of Scraper individuals)/(No.
of filterer individuals)
>50
35-
50%
20-
35%
<20°A
Community Loss
Index(d)
A measure of the dissimilarity
between a test site and a reference
site (Plafkin et al. 1989). Metric
values increase as biological
impairment increase. Values have
no limits.
CLI = (a-b)/c
where: a = no. genera in reference
sample, b = no. genera in test
sample, c = no. genera common to
both samples
<0.5
0.5-1.5
1.5-4.0
>4.0
a) Value is converted to ratio of test to reference site * 100
b) Value is converted to ratio of reference to test site *100
c) Actual percent contribution used in scoring, not ratio to reference
d) Uses range of values actually obtained
6.6
Summary: Determining ALU Support
RI DEM uses a combination of biological, habitat, chemical, and physical data to assign
ALUS (Table 6-3). RI WQS contain some numeric criteria for dissolved oxygen, pH,
temperature and priority pollutants, but other criteria are assessed based on a narrative
description of water quality condition.
Taking into consideration all of the above factors, RI DEM determines the ALU
attainment status of the streams. ALU is fully supported if there are no exceedances of the water
quality criteria and the biological data indicate a fully supporting community. RI DEM gives the
biological component more weight than the water chemistry data for the ALU assessment.
Therefore, a river/stream may be considered fully supporting ALU if the biological community
demonstrates non-impairment or slight impairment, even if minor exceedances of water quality
criteria exist. A stream is considered partially supporting ALU if the macroinvertebrate
assemblage indicates a bioassessment category of "slightly impaired" or "moderately impaired"
and/or if there is an exceedance of any chemical water quality criterion (acute or chronic) more
than once in a three-year period but in fewer than 10% of the samples. A site is determined to be
not supporting ALU if the biological community is "severely impaired" and/or if there are severe
6-6
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Table 6-2. Percent comparability evaluation for macroinvertebrate bioassessment scores
used by the State of Rhode Island.
Bioassessment
Categories
Definition
Percent
Comparability
to Reference
Non-impaired
Comparable to the best condition expected within an
ecoregion. Trophic structure is balanced and community
structure is optimal for the stream size and habitat
quality.
> 83 %
Slightly
impaired
Community structure is less than expected. Species
composition is lower as evident by the loss of some
intolerant species. The percent contribution of tolerant
species increases.
54 - 79 %
Moderately
impaired
Fewer species are present and most intolerant species
disappear.
21 -50 %
Severely
impaired
Few species are present and the stream is often
dominated by one or two species.
< 17 %
or frequent (>10% of samples) violations of chemical water quality criteria. States are required
by the CWA to describe the water quality of their state's waters in the 305(b) report and any
water bodies that are found to be not in attainment of their designated uses must be listed on the
303(d) list. In Rhode Island, any water bodies that are considered partially or not supporting any
designated uses are listed on the State's 303(d) List of Impaired Waters.
Table 6-3. Biolog
(modifiedfrom RI
ical, physical and chemical criteria used to determine aquatic life use
DEM 2000).
Component
Biological
Physical
Chemical
Description of Criteria
• Macroinvertebrate Index (Plafkin et al. 1989)
• Non-impaired, slightly impaired, moderately impaired
severely impaired determined for the stream based on
macroinvertebrate assemblage
, and
the
• Land use evaluation
• Subsystem classification
• Documentation of dams, erosion, and non point source
pollution
• Width and depth measurement
• Flow measurement
• Inorganic and organic substrate types
• Presence of odors, oils and deposits
• RBP Habitat Assessment (Barbour et al. 1999)
6-7
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Component
Description of Criteria
Dissolved • Cold water fish habitat: dissolved oxygen should be above
Oxygen 75% saturation based on a daily average and than the
instantaneous minimum concentration shall be of at least 5
mg/1.
o To protect early life stages that are intergravel during
the fish spawning period of October 1st to May 14th, the
7-day mean water column dissolved oxygen
concentration shall not be less than 9.5 mg/1, while the
instantaneous minimum dissolved oxygen shall not be
less than 8 mg/1.
o To protect those early life stages that are exposed
directly to the water column, the 7-day mean water
column dissolved oxygen concentration shall not be
less than 6.5 mg/1 and the instantaneous minimum
dissolved oxygen concentration shall not be less than
5.0 mg/1.
Warm water fish habitat: daily average dissolved oxygen shall
be above 60% saturation and the instantaneous minimum
dissolved oxygen concentration shall be at least 5.0 mg/1. The
7-day mean water column dissolved oxygen concentration
shall not fall below 6 mg/1.
PH
The pH should be as naturally occurs (6.5-9.0)
Temperature Increase should not rise above the recommended limit that would
cause the growth of nuisance species nor rise above 83°F. In cold
water habitats, heated discharges must not raise the temperature above
68°F and in not case shall the receiving water be raised more than 4°F.
Secchi depth
and chlorophyll
a
Secchi depth and chlorophyll a are used to measure the impact of
nuisance algal blooms that may degrade the quality of life for fish and
wildlife.
Priority
Pollutants
Please refer to
Appendix B of
the Water
Quality
Regulations for
a list of
pollutant
criteria (RI
DEM 2000).
Shall not be found in concentrations or combinations that would be
harmful to humans or fish and wildlife for the most sensitive and
governing water class use, or unfavorably alter the biota, or which
would make the water unsafe or unsuitable for fish and wildlife or
their propagation, impair the palatability of same, or impair waters for
any other existing or designated use.
• Aquatic Life Criteria: The acute and chronic aquatic life
criteria for freshwaters shall not be exceeded at or above the
lowest average 7 consecutive day low flow with an average
recurrence frequency of once in 10 years (7Q10).
• Human Health Criteria: The freshwater human health criteria
for non-carcinogens are applicable at or in excess of the lowest
average 30 consecutive day low flow with an average
recurrence frequency of once in 5 years (30Q5). The
freshwater human health criteria for carcinogens are applicable
at or in excess of the harmonic mean flow, which is a long-
6-8
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Component
Description
term
daily
daily
of Criteria
mean flow value calculated
flows analyzed by the sum
flows.
by dividing the number of
of the reciprocals of those
6.7 Literature Cited
Barbour, M.T., J. Gerritsen, B.D. Snyder, and J.B. Stribling. 1999. Rapid Bioassessment
Protocols for Use in Streams and Wadeable Rivers: Periphyton, Benthic
Macroinvertebrates and Fish, Second Edition. EPA 841-B-99-002. U.S. Environmental
Protection Agency; Office of Water, Washington D.C.
http://www.epa.gov/owow/monitoring/rbp/wp61 pdf/rbp. pdf
Plafkin, J.L., M.T. Barbour, K.D. Porter, S.K. Gross, and R.M. Hughes. 1989. Rapid
Bioassessment Protocols for Use in Streams and Rivers: Benthic Macroinvertebrates and
Fish. EPA 440-4-89-001. U.S. Environmental Protection Agency, Office of Water
Regulations and Standards, Washington, D.C.
(RI DEM) Rhode Island Department of Environmental Management. 2000. Water Quality
Regulations. Regulation EVM 112-88.97-1. Rhode Island Department of Environmental
Management, Office of Water Resources.
http://www.state.ri.us/dem/pubs/regs/REGSAVATER/h20qltv.pdf
RI DEM. 2002a. Quality Assurance Project Plan for Taxonomic Identification of Benthic
Macroinvertebrates, Rhode Island. Rhode Island Department of Environmental
Management, Office of Water Resources.
RI DEM. 2002b. State of Rhode Island and Providence Plantations 2002 Section 305(b) State of
the State's Waters Report. Rhode Island Department of Environmental Management,
Office of Water Resources, http ://www. state.ri .us/dem/pubs/3 05b/index.htm
RI DEM. 2003. Biomonitoring and Habitat Assessment Rhode Island Wadeable Streams 2002
Data Report. Rhode Island Department of Environmental Management, Office of Water
Resources.
6.8 Resources
Rhode Island Department of Environmental Management, Office of Water Resources. Web
Page: http://www.state.ri.us/dem/programs/benviron/water/index.htm
U.S. EPA. Biological Indicators of Watershed Health, Rhode Island Webpage:
http://www.epa.gov/bioindicators/html/state/ri-bio.html
U.S. EPA. 2002. Summary of Biological Assessment Programs and Biocriteria Development or
States, Tribes, Territories, and Interstate Commissions: Streams and Wadeable Rivers.
EPA-822-R-02-048. U.S. Environmental Protection Agency.
6-9
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7 VERMONT
This document was prepared using documents written by the State of
Vermont. Any questions concerning bioassessment methods should
be directed to:
Doug Burnham, Biomonitoring and Aquatic Studies Section
Chief
Vermont Department of Environmental Conservation (VT DEC)
103 South Main Street, ION
Waterbury, VT 05671
Phone: (802) 241-3784; Fax (802) 241-3008
Email: Doug.Burnham@anr.state.vt.us
7.1 Introduction
The Vermont Department of Environmental Conservation (VT DEC) has developed a
biocriteria program for wadeable streams that uses both fish and macroinvertebrate assemblages
for the calculation of indices indicative of stream conditions and biological integrity as it pertains
to ALU classes for 303 (d) listed streams. The foundation of this program was set in 1982 with
the creation of the Ambient Biomonitoring Network (ABN) in the Biomonitoring and Aquatic
Studies Section (BASS) comprised of five biologists focusing on river and stream biological
monitoring. The objectives of the ABN within BASS are to monitor long term trends over time,
evaluate potential impacts from non-point and point sources, establish a reference database
specific to Vermont's classification and attainment determinations, support VT DEC permitting
programs that require data, and conduct special studies to assess emerging water quality and
environmental management issues.
In 1985, VT DEC began using standardized methods to sample fish and
macroinvertebrate communities. These methods included guidelines on evaluating physical
habitat, processing samples, and analyzing and evaluating the data. Vermont now uses an IBI to
measure the condition of the fish community, and multiple metrics applied individually to assess
the integrity of the macroinvertebrate community. These data are then used together to
determine the level of ALUS and attainment for wadeable streams and rivers in Vermont.
Section 3-01 of the VT WQS (Vermont Water Resources Board 2000) states the
provisions to "establish and apply numeric biological indices to determine whether there is full
support of aquatic biota and aquatic habitat uses" and to "establish procedures that employ
standard sampling and analytical methods to characteristics of the biological integrity of the
appropriate reference conditions." VT WQS also lists the water quality management classes and
the biological standards that define the classes. Sections 3-02, 3-03, and 3-04 identify the
management strategies and narrative standards for these water quality management classes.
These classes range from the highest ALUS class Al (ecological or natural conditions) to Class
B Water Management Classes Type 1, 2, and 3, and Class A2 (public water supplies) (Table 7-
1).
7-1
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Table 7-1. Biological ALUS management classes and associated narrative biological
criteria for rivers and streams in Vermont.
Class
Management
Biological Standard
Al
Ecological. Managed to achieve
and maintain waters in a natural
condition. Highest quality
waters.
Reference condition, minimal impacts from
human activity. Highest quality water that
have significant ecological value.
A2
Managed for public water
supplies and to achieve and
maintain waters with uniformly
excellent character and level of
water quality.
High quality aquatic biota and habitat
necessary to support their life cycle and
reproductive requirements.
Moderate change from the reference condition
in the relative proportions of tolerant,
intolerant, taxonomic, and functional
components.
Class B, Water
Management Type 1
Managed so that no change from
the reference condition would
prevent the full support of
aquatic biota, wildlife or aquatic
habitat uses.
Minor change from reference condition, minor
changes in relative proportions of taxonomic
and functional components, relative
proportions of tolerant and intolerant
components within range of reference
conditions. Changes in the aquatic habitat
shall be limited to minimal differences from
the reference condition consistent with the full
support of all aquatic biota and wildlife uses.
Class B, Water
Managements Type 2
Managed so that no change from
the reference condition would
prevent the full support of
aquatic biota, wildlife or aquatic
habitat uses.
Moderate change in the relative proportions of
tolerant, intolerant, taxonomic, and functional
components. Changes in the aquatic habitats
shall be limited to minor differences from the
reference condition consistent with the full
support of all aquatic biota and wildlife uses.
Class B, Water
Managements Type 3
Managed so that no change from
the reference condition would
prevent the full support of
aquatic biota, wildlife or aquatic
habitat uses.
Moderate change in the relative proportions of
tolerant, intolerant, taxonomic, and functional
components. Changes in the aquatic habitats
shall be limited to moderate differences form
the reference condition consistent with the full
support of all aquatic biota and wildlife uses.
When such habitat changes are a result of
hydrological modification or water level
fluctuation, compliance may be determined on
the basis of aquatic habitat studies.
Other Class B Waters
Managed so that no change from
the reference condition would
prevent the full support of
aquatic biota, wildlife or aquatic
habitat uses.
No change from the reference condition that
would have an undue adverse effect on the
composition of the aquatic biota, the physical
or chemical nature of the substrate or the
species composition or propagation of fishes.
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7.2 Key Elements of the Biological Assessment Approach
7.2.1 Index Period and/or Temporal Conditions
The VT DEC has chosen late summer and fall (i.e., September to mid-October) for its
index period to conduct both fish and macroinvertebrate reference and test site sampling.
Samples must be collected during this index period to maintain consistency between the samples
taken from year to year and for accurate community comparisons. This index period was
selected because of the presence of stable, low-flow conditions and because the
macroinvertebrate larvae and fish YOY are larger and easier to identify. Each site is visited one
time during a sampling season.
7.2.2 Monitoring Program Survey Approach
The majority of the streams that are selected for sampling are targeted sites. These sites
encompass specific sampling stations that have been monitored for the purpose of discharge
permitting and TMDLs and can include sites that have been chosen for specific projects.
Additionally sites are selected based on human land use and interest of watershed groups or the
VT DEC planning section's need for data to determine the current biological condition of a
stream reach. Vermont recognizes 17 watershed planning units that are a combination of smaller
sub-basins. Seasonally monitored sites are sampled using a five-year rotating watershed
strategy. This strategy covers three to four basins each year and approximately 125 sites are
sampled per year within these basins (VT DEC 2004).
7.2.3 Natural Classification of Water Bodies
The 17 watershed planning units used by VT DEC were chosen nearly 30 years ago using
the three major watersheds in the state. The three watersheds were then separated into 17
identifiable units. These classifications into 17 units were made based on local habitat features
including elevation, drainage order, stream gradient and substrate composition.
7.2.4 Indicator Assemblages
VT DEC monitors both benthic macroinvertebrates and fish. Benthic macroinvertebrates
are identified to genus or species in the laboratory (Class Oligochaeta is identified to family). A
set of metrics to assess the macroinvertebrate assemblage has been developed using multivariate
statistics. The fish assemblage is identified to species in the field and a multimetric fish index
has been developed for both cold water and mixed water categories.
7.2.5 Reference condition
Reference conditions have been derived from macroinvertebrate and fish assemblage data
independently and take into consideration a range of physical and chemical parameters a
particular community would naturally encounter. The reference sites have been chosen and
defined using the best professional judgment of VT DEC biologists, based on human activity and
the potential that it may affect the stream. Specific reference sites and conditions can be found in
-------
VT DEC (2004). There are three reference condition stream types for macroinvertebrates and
two reference condition stream types for fish, totaling 150 reference sites. If a stream has unique
properties and cannot be compared to one of the reference condition stream types, then a
localized reference stream(s), historic data and best professional judgment are used to determine
the expected natural range of metrics.
Macroinvertebrates reference sites are categorized into one of the following stream types
(VT DEC 2004):
1. Small High Gradient Streams (SHG) are small, first-to-third order headwater streams with
drainage areas averaging 10 km2. These streams are located typically over 1500 ft in elevation
and are highly canopied (83% average canopy cover). These streams have a high gradient and
substrate dominated by gravel/cobble/boulder and approximately 3% fine sediments. Water is
soft and alkalinity would typically measure less than 20 mg/1.
2. Medium Sized High Gradient Streams (MHG) are medium, third to fourth order streams with
a drainage area averaging 88 km2. These streams are found at elevations averaging 814 ft, and
are covered by about 50% canopy. These streams have a high gradient and substrate dominated
by gravel/cobble/boulder and approximately 6% fine sediments. Water has a moderate
alkalinity, typically averaging 48 mg/1.
3. Warm Water Medium Gradient Streams and Rivers (WWMG) are larger streams, fourth to
sixth order, or small streams within the Champlain Valley. Because this category contains larger
streams than those in the Champlain Valley, the drainage areas vary, but the average size is 480
km2. All streams are found at lower elevations averaging 369 ft, are less shaded with an average
30% canopy, and are warmer. These streams have a moderate gradient and substrates dominated
by gravel/cobble/boulder and approximately 7% fine sediments. Water has a high alkalinity,
typically averaging 70 mg/1.
VT DEC categorizes a site by comparing chemical and physical data from a test stream to
those conditions found in a reference stream type. Factors that are taken into consideration when
categorizing a stream are elevation, drainage area, stream order/size, stream gradient, substrate
composition, pH, alkalinity, specific conductance, and other unique characteristics. If these
characteristics at a site are outside the range of those found in the reference sites, then the VT
DEC uses an alternative analysis to describe an appropriate reference condition for site
comparison. This prescription may include using historical monitoring data from the same or
adjacent or similar water body; using a regional reference site; using a site-specific reference site
(e.g., from upstream sites or adjacent sites); using paleo-ecological data collected from the
sediments; or using quantitative models developed from field, historical, and experimental
laboratory data. Any deviation from methods is fully documented by VT DEC and
determination is based on the establishment of a compelling weight-of-evidence argument
derived from monitoring data and best professional judgment.
Fish reference conditions fall into two stream types for which metrics are calculated
(VT DEC 2004):
1. Small cold water streams: A site will be categorized for assessment by the Coldwater Index of
Biotic Integrity (CWIBI) only if the stream can naturally support two to four native, naturally
reproducing species offish. Any wadeable stream site that is located at an elevation greater than
500 ft or within the Connecticut River drainage is classified as a coldwater stream for these
7-4
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purposes. For other streams that are below 500 ft in the Champlain Valley, fish composition will
determine which IBI will be used. Furthermore, the site is classified as cold water if one of the
following is present: one salmonid species, slimy sculpin, or longnose sucker. Those sites that
would naturally meet the cold water criteria but are populated with warm water species as a
result of human influence areas are considered cold water sites for attainment status purposes.
2. Warm water and cold water streams: A site will be placed in this category only if the stream
can naturally support five or more native, naturally reproducing species offish. Stream sites in
this group are evaluated using the Mixed Water Index of Biotic Integrity (MWIBI). Both warm
and coldwater streams fall into this category.
7.3 Field and Laboratory Protocols
7.3.1 Macroinvertebrate Protocols (taken from VT DEC 2004)
7.3.1.1 Field Methods
VT DEC uses a method to collect macroinvertebrates similar to that described in the
Single Habitat RBP III (Plafkin et al. 1989). A riffle habitat is chosen within the stream
sampling reach for macroinvertebrate sampling. Then, an 18-in wide x 12-in high D-frame net
(500 |im mesh) is placed in the riffle and an area immediately upstream of the net (approximately
1.5-ft x 1.5-ft) is thoroughly disturbed by hand, ensuring that all pieces of substrate are moved
and rubbed clean of attached organisms. Moving up-stream, this is repeated at four to five
different locations within the riffle, representing a range of velocity and substrate type
characteristics of that riffle. Each specific location is actively sampled until all the substrate in
approximately an 18-in x 18-in square in front of the net has been disturbed. This generally
takes about 30 seconds of active sampling per location, and active sampling is terminated at the
end of approximately two minutes. The contents of the net are allowed to drain of excess water,
placed into a quart mason jar and preserved with 75% ethanol. Then, to obtain a replicate
sample for the site, this entire process is repeated, being careful to avoid areas previously
disturbed. This "composite" sampling methodology effectively collects samples representative
of the macroinvertebrate assemblage of that riffle. The VT DEC then measures and records the
physical condition of the riffle (i.e., stream wetted and bankfull widths, depth, water velocity,
water temperature, weather conditions, substrate composition, substrate embeddedness, canopy
cover, stream bank condition, and immediate upstream land use) and takes a water sample for
specific conductance, pH and alkalinity determination. On a site-specific basis, other water
chemistry parameters are determined. The site and sampling event codes are recorded on the
field sheet.
7.3.1.2 Laboratory Methods (VT DEC 2004)
Sample processing takes place in the laboratory where macroinvertebrate samples are
washed of preservative through a #30 sieve. The rinsed sample is then spread evenly over a
white gridded tray (minimum 24 squares) by adding a small amount of water to allow the sample
to be evenly spread, but not so much as to cause the macroinvertebrates to float freely around the
tray. Six squares are randomly selected by first choosing one random number and then isolating
7-5
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five surrounding squares from the rest of the sample. This method ensures that one quarter of the
sample collected will be picked. After all macroinvertebrates from one quarter of the tray have
been sorted, additional squares are then sorted until a minimum of 300 organisms has been
reached. The total number of squares sorted is recorded so that sample density or relative
abundance can be calculated. Macroinvertebrates are then identified to genus or species except
for the Class Oligochaeta which is identified to the family level (VT DEC 2004). All
macroinvertebrates are separated into major groups and preserved with 75% ethanol. If the
entire sample is not sorted, it is qualitatively examined for all EPT organisms and other larger
organisms, such as crayfish or mussels, not detected in the subsample to determine species
distributions only. The additional EPT sample is preserved in a separate jar. A reference
collection of all identified taxa is kept to assure consistent identifications.
7.3.2 Fish Protocols (VT DEC 2004)
7.3.2.1 Field Methods
The VT DEC collects fish to determine the community condition of wadeable streams.
Wadeable is defined as a stream or river that at some time during the year can be sampled by an
individual wading in the thalweg of the channel. In streams less than 6 m wide, a single
backpack electrofishing unit is used. In wider streams, the VT DEC uses multiple backpack
units. Reach length is chosen by the overall width of the stream with a minimal reach length of
75 m for a 3-m wide stream, 100 m for a 4- to 5-m wide stream, 120 m for a 6- to 8-m wide
stream, 140 m for a 9- to 11-m wide stream, 160 m for a 12- to 14-m wide stream, and 180 m for
a stream with a width of 15 m. The reach should represent a subsample of the overall stream and
may also be dependent on the expected density of fishes. For example, if an unproductive cold
water stream is sampled, a longer reach is sampled to compensate for the low density offish
expected. Those streams that are productive cool and warm water sites may have smaller sample
reaches. Furthermore, the stream section that is sampled would represent the overall habitat of
the surrounding stream reach. All habitat types are sampled within the reach to maximize
species richness.
When electrofishing, the crew begins in the most downstream portion of the reach and
moves upstream. If a stream is being screened, only one pass is required. Two or three passes
are made in those streams where the density is being evaluated as a result of a specific impact. If
multiple passes are made, the VT DEC calculates a removal population estimate to ensure the
accurate calculation offish density. All fish that are stunned are collected using a net and placed
into buckets of water for on-site identification to species and then released. All fish are examined
for anomalies and salmonids are measured for length. Generally all identifications occur in the
field. If a positive identification cannot be made, the specimen is preserved and is positively
identified by a VT DEC biologist using keys (Smith 1985, Langdon et al. in preparation).
7.4 Data Management/Quality
Data are stored and managed in a Microsoft Access database, and metrics are calculated
in this program. Data stored in the Access database can be moved to other spreadsheet programs
such as Excel and to graphical and statistical analysis programs such as SigmaPlot, SigmaStat,
7-6
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and PC-ORD. VT DEC uses SigmaStat for the statistical analysis calculations of parametric
ANOVAs and nonparametric comparisons, and uses PC-ORD for multivariate analyses.
7.5
Analysis of Biological Data
7.5.1 Macroinvertebrate Data (VT DEC 2004)
VT DEC uses multivariate statistics and multiple metrics to evaluate the
macroinvertebrate assemblage. Eight metrics have been developed to measure the
macroinvertebrate community integrity (Table 7-2). These metrics measure specific ecological
attributes of the community and include: Density, Richness, EPT index, Percent Model Affinity
Orders, HBI, Percent Oligochaeta, EPT/(EPT + Chironomidae), and the Pinkham-Pearson
Coefficient of Similarity-Functional Groups (Table 7-2). Using macroinvertebrate data, each
metric is scored and an overall condition rating is determined for each site sampled.
After metrics are calculated, the values are compared to a table of threshold values for each of
the water quality classes (Table 7-3). These values represent a single point in a continuum of
values reflecting changes from the reference condition. Along with best professional judgment
and a weight-of-evidence decision process, VT DEC assigns a site to an attainment class. The
biologist assigns a pass, fail, or indeterminate value for each of the metrics depending on the
score. If five or more metrics score a pass and no metrics are below the threshold value, then the
ALU is supported. However, if one or more of the metrics fail the ALU is not supported. If
neither of these conditions is met for the Water Management Class of the stream being assessed
(i.e., Class Al or B), then the site is assigned an indeterminate finding and may require
additional data and/or sampling to make a support or non-support decision.
Table 7-2. VT DEC macroinvertebrate metrics and methods used to calculate each of the
metrics.
Variable
Number
Indices/ Measures of
Biological Integrity
Method
1
Density
The relative abundance of animals in a sample. (Number of animals
in subsample)/(Proportion of sample processed)
Richness
The number of species in a sample unit. Calculated as the total
number of distinct taxa identified in a sample and averaged across
replicate samples. Note: immature larva identified to family or
genus is not considered new taxon if genus or species identification
is determined within its group.
EPT Index
The number of distinct taxa identified in a sample from the orders
Ephemeroptera, Plecoptera, and Trichoptera. Calculated as for
Variable 2. Note: immature larva identified to family or genus is not
considered new taxa if genus or species identification is determined
within its group.
7-7
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Variable
Number
Indices/ Measures of
Biological Integrity
Method
Percent Model Affinity
Orders (PMA-O)
A measure of order-level similarity to a model based on the reference
streams. PMA-O is calculated by determining the percent
composition for each major group (Coleoptera, Diptera,
Ephemeroptera, Plecoptera, Trichoptera, and Oligochaeta) at the
assessment site. Then those values are compared to the mean
percent composition of each corresponding order from the reference
condition (model). The sum of the minimum of the two values for
each order is the PMA-O.
PMA-O = E min (Xa or Xr)
Where: Xa= the percent composition of order X from the assessment
site
Xr = the percent composition of order X from the appropriate
reference conditions (Novak and Body 1992).
Hilsenhoff Biotic Index
(HBI)
N
Where:
Hi = number of individuals in taxon /
at = tolerance value for taxon /' (as assigned by VT DEC 2004, after
Hilsenhoff 1987, Bode et al. 1996)
N = number of individuals in the sample with a tolerance value
Percent Oligochaeta
(Abundance of Oligochaeta)/(Total number of individuals) x 100
EPT/EPT +
Chironomidae
(Abundance of Ephemeroptera, Plecoptera and Trichoptera) /
(Abundance of Ephemeroptera, Plecoptera, Trichoptera +
Chironomidae)
Pinkham-Pearson
Coefficient of Similarity-
Functional Groups
(PPCS-F)
A measure of functional feeding groups calculated by determining
the percent composition of the six major functional groups
(collector-gatherer, collector-filterer, predator, shredder-detritus,
shredder-herbivore, scraper) at the site as assigned by VT DEC
(2004) (based onMerrit and Cummins 1996, Bode et al. 1996).
l=l max(xia,xlb)j
Where:
K = the number of comparisons between stations (6)
xia = the number of individuals in functional group /' in sample a
(reference site)
xib = number of individuals in functional group /' in sample b
The final decision is based on a number of considerations in accordance with the VT
WQS. These include the use of chemical and physical data from the sample site to determine
which of the three wadeable stream macroinvertebrate categories are used to determine
7-S
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attainment, as well as the evaluation of data quality. Macroinvertebrate data must be collected
using standard methods and trained qualified personnel. Furthermore, any diversions from
average conditions must be documented by VT DEC. These diversions could include any
hydrological, meteorological or other extreme events that occurred before sampling, and any
errors committed during sampling. Finally, variability of the samples from a site is analyzed and
evaluated on a case-by-case basis. If variability exceeds 40%, but is less than 75% the data will
be handled with caution. If the percent standard error of the mean of abundance is greater than
75%, the data are rejected. If this is the case, any other data collected from the site (e.g.,
physical, chemical, and/or fish data) can be used to submit a temporary ALUS classification.
VT DEC then sets threshold index values to define how the biological indices relate to
the narrative class standards established in the VT WQS. The threshold values were derived
from the distribution of metrics within both the reference and the impacted data sets. Thresholds
are also minimally adjusted at each site on a case-by-case basis by a VT DEC biologist using
BPJ.
Class A includes only those highest quality sites that exhibit minimal change from natural
conditions. For those metrics that decrease in value with impairment from stressors (i.e.,
Richness, EPT, PMA-O, EPT/ EPT +C, PPCS-FG, and Density), it is reasonable to expect that
the upper 75 percent of the reference sites best meet the true natural condition (threshold set at
25th percentile). For the metrics that increase in value with impairment from a stressor (i.e., HBI
and percent oligochaetes) the threshold was set at the 75l percentile.
Streams in Class B, Water Management Type 1 exhibit minor changes from "natural
condition". The reference streams are likely include a small percentage of lower quality
reference sites; therefore, the threshold values for Class B1 were set to the 5th and 95th percentiles
of the reference streams to ensure against the influence of outlier reference values. Since a
minor change was expected for these sites, the threshold was set at the 5th percentile (to include
the upper 95 percent of the reference sites) for the metrics that decrease in response to stress
(e.g., Richness). For metrics that increase in response to stress, the threshold was set at the 95th
percentile of reference sites, such that most reference sites fell below the threshold.
Class B Water Management Types 2 and 3 and Class A2 allow a moderate change from
"natural condition" as a management goal. Thresholds for these classes were set based on BPJ
using the range of reference values and the median, and I0ih/90ih percentile values of the
distribution from sites known to be impacted.
7.5.2 Fish Data (VT DEC 2004)
The fish assemblage at sampling sites is assessed using either the Cold Water Index of
Biotic Integrity (CWIBI) or the Mixed Water Index of Biotic Integrity (MWIBI). VT DEC
derived index values consistent with narrative biological criteria in VT WQS using data from
reference and impaired sites. The CWIBI consists of six metrics and is applied to small cold
water stream fish communities, while the MWIBI consists of nine metrics and is applied to cold
or warm water communities in Vermont. Tables 7-4 and 7-5 list the metrics, calculations, and
scoring criteria for the CWIBI and MWIBI, respectively. The scoring criteria were derived for
the CWIBI (7.5, 4.5, and 1.5) and the MWIBI (5, 3, and 1) to compare the cold water and mixed
water sites with each other. For example, there are six CWIBI metrics and the highest overall
score possible is 45 (7.5 x 6 = 45), whereas there are nine MWIBI metrics and the highest overall
score possible is also 45 (9 x 5 = 45). Each metric is scored based on definitions and habitat or
7-9
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basin conditions. A final summation of each of the scores is made to place a fish assemblage
(i.e., site) into a condition category (i.e., Excellent, Very Good, Good, Fair, Poor). "Excellent" is
assigned to those streams that achieve a score of at least 41, "Very Good" to streams with scores
of at least 36, "Good" for scores of at least 33, "Fair" for scores of at least 27, and anything
below 27 is assigned a condition of "Poor". The CWTBI or MWTBI score is then compared to
the range of scores that correspond to each of the VT WQS Classes to determine which class
could potentially best describe the fish assemblage (Table 7-6). For example, if a cold water fish
assemblage scores a 36 on the CWIBI, the stream could be classified B Water Management Type
1, defined as exhibiting a minor change from the reference condition.
As is the case with macroinvertebrate data, fish data must undergo a stringent evaluation.
Qualified personnel must adhere to the SOPs when collecting and analyzing the data. If a
sample is deemed unacceptable due to error or a unique event other than what occurs in an
average year, VT DEC can sample the site again. This event is to be scheduled at least three
weeks from the initial sampling date. Until new data are collected, the VT DEC can assign a
temporary attainment value based on the valid biological, chemical, and physical data already
collected.
7.6 Summary: Determining ALU Support
VT DEC calculates metrics for both macroinvertebrate and fish data collected from probabilistic
and monitoring stations for use in determining ALU attainment. Although each of the metrics
and indices were derived independently, they each provide a quantitative way to assess the
biological condition in the diverse types of streams in Vermont. VT WQS provide a
classification system describing the condition of these streams (Table 7-1) that include Class Al,
A2 and Class Bl, B2, and B3 streams. Once these classes were established with management
provisions in place, indices were developed that describe the stream condition in relation to
reference condition site values (e.g., CWIBI, MWIBI and macroinvertebrate metrics). The VT
DEC uses the CWIBI, MWIBI and individual macroinvertebrate metrics to determine the ALUS
based on the thresholds described in the tables above (Tables 7-3, 7-4, 7-5, and 7-6). The
resultant product is a narrative description for each monitored site for the ALU attainment part of
the 305(b) report on the condition of streams in Vermont. In addition, this information is used to
list non-supporting sites on the 303(d) list for Vermont.
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Table 7-3. Macroinvertebrate assemblage biocriteria thresholds for the macroinvertebrate community stream categories, and
associated WQ classes of Vermont (VT DEC 2004).
WQ Class
Metric
Richness
EPT
PMA-O
HBI
% Oligo
EPT/EPT
+Chiron.
PPCS-FG
Density
Direction of
metric as
water
quality
improves
Positive
Positive
Positive
Negative
Negative
Positive
Positive
Positive
Small-size High Gradient Streams
(SHG)
Al
Ecological
Reference
Condition
>35
>21
>65
<3.00
<2
>0.65
>0.50
>500
B-WMT1
Minimal
Change
from
Reference
Condition
>31
>19
>55
<3.50
<5
>0.55
>0.45
>400
B
B-WMT 2-3
A2 (water
supply)
Moderate
Change from
Reference
Condition
(undue adverse
effect)
>27
>16
>45
<4.50
<12
>0.45
>0.40
>300
Medium-size High Gradient Streams
(MHG)
Al
Ecological
Reference
Condition
>43
>24
>65
<3.50
<2
>0.65
>0.50
>500
B-WMT1
Minimal
Change
from
Reference
Condition
>39
>22
>55
<4.00
<5
>0.55
>0.45
>400
B
B-WMT 2-3
A2 (water
supply)
Moderate
Change from
Reference
Condition
(undue adverse
effect)
>30
>18
>45
<5.00
<12
>0.45
>0.40
>300
Warm Water Medium Gradient Streams
and Rivers (WWMG)
Al
Ecological
Reference
Condition
>40
>21
>65
<4.25
<2
>0.65
>0.50
>500
B-WMT1
Minimal
Change
from
Reference
Condition
>35
>19
>55
<4.75
<5
>0.55
>0.45
>400
B
B-WMT 2-3
A2 (water
supply)
Moderate
Change from
Reference
Condition (undue
adverse effect)
>30
>16
>45
<5.40
<12
>0.45
>0.40
>300
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Table 7-4. The six metrics used in scoring the fish assemblage for the CWIBI. These
streams must naturally support two to four native species (VT DEC 2004).
Metrics and qualifications/calculations
1. Number of intolerant species
(One exotic trout species may be substituted for brook trout)
2. Proportion of individuals as cold water stenotherms
3. Proportion of individuals as generalist feeders
4. Proportion of individuals as top carnivores
5. Brook trout density (number/100 m2 - Ipass)
6. Brook trout age class structure
Young-of-the-year (YOY). 100 mm, adult. 100mm)
Index Scores
Excellent 42-45
Very Good 36
Good 33
Fair 27
Poor < 27
Scoring Criteria
7.5
2
> 75%
<5%
> 35%
>4.0
YOY and
adults present
4.5
1
50-75%
5-9%
25-35%
2.0-4.0
YOY only
1.5
0
< 50%
>9%
< 25%
<2.0
YOY absent
Conditions for use
1. Only fishes over 25 mm in length should be considered.
2. Only naturally reproducing salmonids are to be considered; no stocked fish are to be
included. No Atlantic salmon are to be included.
3. Only species represented by more than a single individual are entered into metrics 1
and 6.
4. No non-resident species shall be entered into metric calculations.
Table 7-5. The nine metrics used in scoring cold and warm water sites for the MWIBI.
These streams must naturally support more than four native fish species (VT DEC 2004).
Metric and Qualification
Stream descriptor
Species Richness and Composition
1. Total number of native fish species
2. Number and identity of native,
intolerant species
(A non-native trout may be substituted for
brook trout when absent)
3. Number and identity of native benthic
insectivores
4. Proportion of individuals as white
suckers and creek chubs.
Site Elevation > 400 feet
Site Elevation < 400 feet
Site Elevation < 400 feet
with site drainage < 25
km2
All other sites
Trophic Composition
Scoring Criteria
5
3
1
Follows maximum species
richness lines
>1
>0
>0
>1
<11%
1
-
-
1
11-30%
0
0
0
0
> 30%
7-12
-------
Metric and Qualification
5. Proportion of individuals as generalist
feeders.
6. Proportion of individuals as water
column and benthic insectivores (score a
"1" if blacknose dace is >60% of total
assemblage or 100% of insectivores)
7. Proportion of individuals as top
carnivores
(Normative trout included)
Stream descriptor
Scoring Criteria
Site Elevation > 500 feet < 20% 20-45% >45%
Site Elevation < 500 feet
Site Elevation > 500 feet
Site Elevation < 500 feet
Cold water assemblage
Warm water assemblage
with site drainage > 25
km2
Warm water assemblage
with site drainage < 25
km2
Fish Abundance and Condition
8. Proportion of individuals with
deformities, fin erosion, lesions, or
tumors.
9. Abundance in sample
(One pass - # 100 m2)
(Nonnative species included)
* Site automatically scores Poor
Sum of Metric Scores
Excellent 41-45
Very Good 37
Good 33
Fair 27
Poor <27
Site Elevation < 500 feet
Site Elevation > 500 feet
Alkalinity > 9 mg/1
Site Elevation < 500 feet
Alkalinity < 9 mg/1
< 30%
> 65%
> 55%
> 15%
> 10%
>o
<1%
>20
>10
>6
30-60%
30-65%
20-55%
5-15%
3-10%
-
1-4%
10-20
7-10
3-6
> 60%
< 30%
< 20%
<5%
<3%
-
>4%
< 10*
<7*
<3*
Conditions for Use
1. For wadeable streams only.
2. Site should naturally support at least five native
species.
3 . Only individuals more than 25 mm TL are to be
entered in the determination.
4. Only species with more than one individual captured
are entered into metrics 2 and 3.
5. Stocked fish are not considered in determinations.
6. All sites within the Connecticut River drainage are to
be scored as > 500 m elevation.
7-13
-------
Table 7-6. All possible scores for the CWIBI and MWIBI that correspond to the VT WQS
classification scheme (VT DEC 2004).
WQS Classification
A-l
A-l orB-lbasedonBPJ
B-l
B-l, A-2, or B-2, B-3 based on BPJ
A-2, B-2, B-3
B-2, B-3 or Non-Support based on BPJ
Non-Support
Range
41-45
39
36-37
35
33
29-31
<29
Possible Scores
CWIBI
42,45
39
36
33
30
27,24,21, 18,
15, 12,9
MWIBI
41,43,45
39
37
35
33
31,29
27,25,23,21,
19, 17, 15, 13,
11,9
7.7
Literature Cited
Bode, R.W., M.A. Novak, and L.E. Abele. 1996. Quality Assurance Work Plan for Biological
Stream Monitoring in New York State. New York State Department of Environmental
Conservation, Albany, NY.
Hilsenhoff, W.L. 1987. An improved biotic index of organic stream pollution. Great Lakes
Entomologist 20:31-39.
Langdon, R.W., M. Ferguson, and K. Cox. In preparation. The Fishes of Vermont.
Merrit, R.W., and K.W. Cummins (editors). 1996. An Introduction to Aquatic Insects of North
America, 3rd ed. Kendall/Hunt Publishing Company, Dubuque, IA.
Novak, M.A., and R.W. Bode. 1992. Percent model affinity, a new measure of
macroinvertebrate community composition. Journal of the North American
Benthological Society 1 l(l):80-85.
Plafkin, J.L., M.T. Barbour, K.D. Porter, S.K. Gross, and R.M. Hughes. 1989. Rapid
Bioassessment Protocols for Use in Streams and Rivers: Benthic Macroinvertebrates and
Fish. EPA 440-4-89-001. U.S. Environmental Protection Agency, Office of Water
Regulations and Standards, Washington, D.C.
Smith, C.L. 1985. The Inland Fishes of New York State. New York State Department of
Environmental Conservation, Albany, NY.
7-14
-------
U.S. EPA. 2002. Summary of Biological Assessment Programs and Biocriteria Development
for States, Tribes, Territories, and Interstate Commissions: Streams and Wadeable Rivers.
EPA-822-R-02-048. U.S. Environmental Protection Agency.
State of Vermont. 2000. Vermont Water Quality Standards. State of Vermont, Water
Resources Board, Montpelier, VT. http://www.state.vt.us/wtrboard/july2000wqs.htm
(VT DEC) Vermont Department of Environmental Conservation. 2004. Biocriteria for Fish and
Macroinvertebrate Assemblages in Vermont Wadeable Streams and Rivers -
Implementation Phase. Vermont Department of Environmental Conservation,
Waterbury, VT. http://www.vtwaterquality.org/bass/htm/bs_biomon.htm
7.8 Resources
U.S. EPA. Biological Indicators of Watershed Health, Vermont Webpage:
http://www.epa.gov/bioindicators/html/state/vt-bio.html
U.S. EPA. (2002). Summary of Biological Assessment Programs and Biocriteria Development
for States, Tribes, Territories, and Interstate Commissions: Streams and Wadeable Rivers.
EPA-822-R-02-048. U.S. Environmental Protection Agency.
http://www.epa.gov/bioindicators/html/program_summary.html
VT DEC, Water Quality Division, Biomonitoring and Aquatic Studies Web page:
http://www.vtwaterqualitv.org/bass.htm
7-15
-------
8 SUMMARY
8.1 Comparison Across States
The U.S. EPA allows for each state to implement its own bioassessment program. This
document reviews the methods used by U.S. EPA Region 1 states to monitor and assess streams
for ALU attainment for 305(b) reporting and 303(d) listing as required by the CWA. Although
Region 1 states share many commonalities such as the Connecticut River, which traverses four
of the six states (New Hampshire, Vermont, Massachusetts, and Connecticut) and has a
watershed that covers 11,000 mi2, the entire region has diverse habitats ranging from northern
broadleaf forests to southern New England forests, coastal systems, northern conifer forests and
alpine regions (Alden and Cassie 2000). The diverse topography causes distinct differences in
biota and, therefore, requires states to make adjustments to their bioassessment programs to
account for these differences. For example, although 41 entities use the fish assemblage for
bioassessment, Maine does not assess fish (U.S. EPA 2002b). One reason Maine has chosen to
assess benthic macroinvertebrates and not the fish assemblage as well, is due to the great
diversity of this assemblage in state waters compared to the fish assemblage (Personal
Communication, Susan Davies). Meanwhile, Vermont has diverse benthic macroinvertebrate
and fish fauna and they have adapted their bioassessment program to include a benthic metrics
and indices to assess cold water and warm water fish assemblages. The rationale for program
development can stem from a biological point of view but may be influenced by historical
sampling methods, feasibility of sampling protocols, and by funding available to the program.
Due to regional similarities, some states share many of the same protocols and have borrowed
ideas for their own index development. Summary Table 8-1 offers a comparison and contrast of
the key components of the state bioassessment programs and Table 8-2 lists all of the metrics
used by the states.
States are responsible for setting narrative criteria that will protect waters and define
ALU. Bioassessment programs are then designed to aid in the determination of ALU using
biological, physical, chemical and habitat data. Biological data provide the most accurate
information about the resident aquatic organisms (U.S. EPA 2002a); therefore, it is essential that
appropriate indicator assemblages be chosen to properly assess the natural system. All U.S. EPA
Region 1 states minimally have chosen to assess the benthic macroinvertebrate assemblage, but
some attempt is being made by most of the states to at least include information about fish
assemblages (i.e., Connecticut, Massachusetts, New Hampshire, and Vermont) and/or algae (i.e.,
Connecticut and Massachusetts). It is then the responsibility of the states to decide the best way
to analyze the biological data in order to show biotic gradients as water quality changes.
As apparent in Table 8-1, these programs do share some commonalities such as using
modified methods from the U.S. EPA RBP document, although the details of the methods may
differ considerably. These differences are evident at all levels, including the strategy used for
site selection, the decision making process used to select reference sites, field collection
methods, laboratory sorting and taxonomic methods, and finally the selection of metrics and data
analysis. These differences could be due to the necessity to compare to historical data by using
similar methods, topographical or regional differences, available equipment, logistical
constraints, and funding provided to the states.
J-l
-------
Table 8-1. Comparisons of the key com
Component
Index Period
Indicator
assemblages used
Survey approach
Total stream miles
# sites sampled/year
Reference condition
CONNECTICUT
Late fall
Benthic
Fish
Periphyton
5-yr rotating basin;
Targeted. CTinthe
process of
developing a
probabilistic
component to
monitoring program
5,830 miles
Benthic-50 sites
Fish- 24 sites
Least disturbed sites
comparable to test
sites based on natural
features such as
gradient
ponents of state bioassessment programs.
MAINE
July-September
Benthic
5-yr rotating
basin; Targeted
31,672 miles
50-60 sites
BPJ for a priori
placement into
four pre-defined
classes for
linear
discriminant
analysis
MASSACHUSETTS
July-September
Benthic
Fish
Periphyton
5-yr rotating basin;
Targeted
8,229 miles
75 sites
Least impacted sites; no
potential to receive
point or non-point
source pollution and
lack land use patterns
that would degrade
water quality
NEW
HAMPSHIRE
mid summer-
early fall
Benthic
Fish
Targeted within
Northern and
Southern regions
10,881 miles
25-30 sites
Least impacted
sites based on
Human
Disturbance
Gradient,
reference sites
within each
bioregion used
to set attainment
thresholds
RHODE
ISLAND
Summer and
fall
Benthic
Fixed stations
within two
Level IV
Omernick
ecoregions
(Adamsville
Brook and
Wood River)
1,498 miles
45 sites
2 least
disturbed
reference sites,
one in each of
the ecoregions
VERMONT
Late summer to
fall
Benthic
Fish
5-yr rotating
basin; Targeted
7,099 miles
125 sites
BPJ used to
identify
reference sites
for 3 stream
types for
benthos, 2 types
for fish
Macroinvertebrates
Sampling approach
RBP III Single
Habitat
(Plafkin et al. 1989)
Classification
Attainment
Evaluation
(Davies et al.
1999, Davies
and Tsomides
RBP II and III Single
Habitat Approach
(Plafkin etal. 1989)
NHDES
methods
(NH DBS 2004)
RBP III Single
Habitat
(Plafkin et al.
1989)
modification of
RBP III Single
Habitat (Plafkin
etal. 1989)
8-2
-------
Component
Field method(s)
Lab sorting method
Organisms
subsample count
Taxonomic
resolution
Analysis method
# of metrics (See
Table 8-2 for
metrics)
CONNECTICUT
Kick net 9-in x 18-in
rectangle, 800 x 900
(im mesh
Entire sample over a
56-square grid
200
genus/species or
lowest practical
taxonomic level
RBPIII metrics and
index
(Plafkin et al. 1989)
7
MAINE
2002)
Rock baskets,
riffle bags, rock
filled cones.
Deployed for
28-56 days. 600
(im mesh sieve
Entire sample
in small
quantities
Entire sample,
unless >500
organisms in
sample
genus/species
or lowest
practical
taxonomic level
Linear
discriminant
analysis
25
MASSACHUSETTS
Kick net 0.46 m x 0.46
m, 500 (am mesh.
Rock baskets deployed
for 6 to 8 weeks.
RBP II and III (Plafkin
etal. 1989)25 squares
6-cm x 6-cm
100
genus/species or lowest
practical taxonomic
level
RBP II and III metrics
and index (Plafkin et al.
1989)
9
NEW
HAMPSHIRE
Rock baskets
deployed for 8
weeks, 600 (im
mesh
Caton method
(Barbour et al.
1999) using 16
square grids.
minimum of 100
genus/species or
lowest practical
taxonomic level
B-IBI for
northern and
southern regions
(Neils and
Blocksom 2004)
7
RHODE
ISLAND
0.3 m wide D-
frame net, 500
(im mesh
18-in x 13-inx
1-in gridded
tray with 8
equal squares
100
genus/species
or lowest
practical
taxonomic
level
RBPIII
metrics and
index (Plafkin
etal. 1989)
11
VERMONT
18-in x 12-inD-
frame net, 500
(im mesh
24-square grid
minimum of 300
Genus or
species,
oligochaetes to
family
Multiple metrics
(VT DEC 2004)
8
Periphyton
Current Use
Developing an algal
indicator using
probabilistic
monitoring; currently
used only for
Not Used
Used for the detection
of algal blooms and as
indicator of water
quality to identify
toxicity issues, nutrient
Not Used
Not Used
Not Used
8-3
-------
Component
Field method(s)
Measurements taken
Use for attainment
CONNECTICUT
supplementary
information.
Modified RBP
Single Habitat
method,
field-based Rapid
Periphyton Survey
(viewing bucket)
(Barbour et al. 1999)
Chlorophyll a,
biomass, species
composition and
abundance
Literature metrics to
derive conclusions
MAINE
MASSACHUSETTS
impacts, and habitat
alterations.
RBP Single Habitat
method, artificial
substrates (biomass and
chl a), and viewing
bucket (% coverage)
(Barbour et al. 1999)
Chlorophyll a, biomass,
and percent coverage
Use to evaluate if the
aesthetics or Aquatic
Life Use are affected
NEW
HAMPSHIRE
RHODE
ISLAND
VERMONT
Fish
Field method
Analysis method
RBP V protocols
(Plafkin et al. 1989):
electrofish 150 m
reach, fish identified
to species in field
Currently developing
an index modeled
after Vermont; data
collected currently
used as
supplementary
information for ALU
determination
Not Used
RBP V protocols
(Plafkin et al. 1989):
electrofish 100 m
reach, fish identified to
species in field
Modification of RBP V
metrics (Plafkin et al.
1989)
RBP V protocols
(Plafkin et al.
1989):
electrofish 150
m reach, fish
identified in
field
IBI under
development and
modeled after
Vermont. In the
process of
refining CWIBI
and MWIBI for
NH stream fish
communities
Not Used
Electrofish a
minimum reach
of 75 m, reach
length varies
with stream
width, usually
identified in
field
CWIBI for cold
water stream
fish
communities,
MWIBI applied
to cold or warm
water
communities
(VT DEC 2004)
8-4
-------
Component
Use offish data
ALUS
determination: data
used
CONNECTICUT
Fish Species
composition, trophic
structure and age
class distribution
along with BPJ used
to make assessment
Biological
(macroinvertebrate
quantitative index,
supplemental fish
data) physical,
chemical,
toxicological and
habitat data also used
MAINE
Heaviest weight
on biological
data (outcome
of the benthic
linear
discriminant
analysis),
physical,
chemical,
bacterial, and
habitat
MASSACHUSETTS
Structure and function
of the fish assemblage
and BPJ used to
determine support or
impairment of aquatic
life use
Weight of Evidence
approach using
biological, habitat,
chemical, and
toxicological data
NEW
HAMPSHIRE
Currently
supplements
macroinvertebrat
e and other data
for attainment
decisions
Heavy weight
placed on
biological (B-
IBI) data, with
fish assessments,
benthic deposits,
flow, habitat
(RBP),
macrophyte
composition,
sediment and
ambient water
toxicity tests, pH
and DO
RHODE
ISLAND
Biological
(macroinverte
brate
community
score),
physical,
chemical (DO,
pH,
temperature),
and habitat
(RBP) data
VERMONT
Used to make
attainment
decisions for
ALUS
Heaviest weight
on biological
(benthic metrics
and fish CWIBI
and MWIBI)
data, with
physical,
chemical, and
habitat data
-------
Table 8-2. Comparison of the macroinvertebrate metrics used by states in the New England Region. Color shading indicates
equivalent metrics across states.
CONNECTICUT
Community Loss
MAINE
Cheumatopsyche
Mean Abundance
Ratio of Class A
Indicator Taxa
MASSACHUSETTS
Community Loss
NEW
HAMPSHIRE
Percent
Chironomidae
RHODE
ISLAND
Community Loss
VERMONT
Density
EPT Index
Chironomini Mean
Abundance (Family
Functional Group)
Ratio of EP
Generic Richness
EPT Index
Percent
Clingers
EPT Abundance
EPT Index
EPT/Chironomidae
(abundance ratio)
Ephemeroptera
Mean Abundance
Relative
Chironomidae
Abundance
~T
(a
PT/Chironomidae
(abundance ratio)
Percent
Intolerant
EPT Taxa
Richness
EPT/EPT +
Chironomidae
Percent Model
Affinity Orders
(PMA-O)
EPT Generic
Richness
Relative Diptera
Richness
HBI/FBI
Percent Non-
insects
EPT/Chironomid
ae(abundance
ratio)
HBI
Percent
Contribution of
Dominant Taxon
EPT-Diptera
Richness Ratio
Relative
Ephemeroptera
Abundance
Percent Contribution
of Dominant Taxon
Percent
Tolerant taxa
HBI
Scraper/Filtering
Ratio
Taxa Richness
Generic Richness
HBI
Relative
Oligochaeta
Abundance
Relative Plecoptera
Richness
Percent Reference
Affinity
Plecoptera
Taxa
Percent
Contribution of
Dominant Taxon
Percent Similarity
Total Taxa
Percent
Hydropsychidae
of Total
Trichoptera
Pinkham-
Pearson
Coefficient of
Similarity-
Functional
Groups (PPCS-
I)
Hydropsyche Mean
Abundance
Shannon-Weiner
Generic Diversity
Scraper/Filtering
Ratio
Ratio of
Shredders to
Total Number of
Individuals
Richness
8-6
-------
CONNECTICUT
MAINE
Perlidae Mean
Abundance (Family
Functional Group)
Plecoptera Mean
Abundance
Probability (A + B
+ C) from First
State Model
Probability (A + B)
from First Stage
Model
Probability of Class
A from First Stage
Model
Sum of Mean
Abundances of:
Cheumatopsyche ,
Cricoptopus,
Tanytarsus and
Ablabesmyia
Sum of Mean
Abundances of:
Dicrotendipes,
Microspectra,
Parachironomus
and Helobdella
Sum of Mean
Abundances of:
Acroneuria and
Stenonema
Tanypodinae Mean
Abundance (Family
Functional Group)
Total Mean
Abundance
MASSACHUSETTS
Taxa Richness
NEW
HAMPSHIRE
RHODE
ISLAND
Scraper/Filtering
Ratio
Shannon Weaver
Diversity Index
Total Taxa
Richness
VERMONT
5-7
-------
Currently, U.S. EPA Region 1 is conducting the New England Wadeable Streams
(NEWS) Project. The primary purpose of this study is to apply a random probability-based site
selection strategy across New England states. As part of this study, fish, macroinvertebrate,
water chemistry, physical chemistry, and habitat data are being collected at 50 sites in each
participating state, either by the state itself or by Region 1. In some cases, individual states are
collecting samples using both the standardized method for this study and their own method,
allowing for a possible comparison of field sampling methods. Sampling was to be completed
by the end of 2003, and a final report presenting the findings of this study is anticipated by late
2004 (Personal Communication, Hillary Snook, U.S. EPA Region 1).
8.2 Literature Cited
Alden, P., and B. Cassie. 1998. The National Audubon Society Field Guide to New England.
Alfred A. Knopf, New York.
Barbour, M., J. Gerritsen, B.D. Snyder, and J.B. Stribling. 1999. Rapid Bioassessment
Protocols for Use in Streams and Wadeable Rivers: Periphyton, Benthic
Macroinvertebrates and Fish, Second Edition. EPA 841-B-99-002. U.S. Environmental
Protection Agency, Office of Water, Washington, D.C.
http://www.epa.gov/owow/monitoring/rbp/wp61 pdf/rbp. pdf
Davies, S.P., L. Tsomides, J.L. DiFranco, and D.L. Courtemanch. 1999. Biomonitoring
Retrospective: Fifteen Year Summary for Maine Rivers and Streams. Maine Department
of Environmental Protection.
http://www.state.me.us/dep/blwq/docmonitoring/biomonitoring/biorep2000.htm
Davies, S.P., and L. Tsomides. 2002. Methods for the Biological Sampling and Analysis of
Maine's Rivers and Streams. Maine Department of Environmental Protection, Augusta,
ME. http://www.state.me.us/dep/blwq/docmonitoring/finlmethl.pdf
Neils, D., and K. Blocksom. 2004. Development of the New Hampshire Benthic Index of Biotic
Integrity. New Hampshire Department of Environmental Services, Concord, NH.
New Hampshire Department of Environmental Services. 2004. Biomonitoring Program
Protocols. New Hampshire Department of Environmental Services, Concord, NH.
Plafkin, J.L., M.T. Barbour, K.D. Porter, S.K. Gross, and R.M. Hughes. 1989. Rapid
Bioassessment Protocols for Use in Streams and Rivers: Benthic Macroinvertebrates and
Fish. EPA 440-4-89-001. U.S. Environmental Protection Agency, Office of Water
Regulations and Standards, Washington, D.C.
(U.S. EPA) U.S. Environmental Protection Agency. 2002a. Consolidated Assessment and
Listing Methodology, Toward a Compendium of Best Practices, First Edition. U.S.
Environmental Protection Agency, Office of Water, Washington, D.C.
http://www.epa.gov/owow/monitoring/calm.html
-------
U.S. EPA. 2002b. Summary of Biological Assessment Programs and Biocriteria Development
for States, Tribes, Territories, and Interstate Commissions: Streams and Wadeable Rivers.
EPA 822-R-02-048. U.S. Environmental Protection Agency.
Vermont Department of Environmental Conservation. 2004. Biocriteria for Fish and
Macroinvertebrate Assemblages in Vermont Wadeable Streams and Rivers -
Implementation Phase. Vermont Department of Environmental Conservation,
Waterbury, VT. http://www.vtwaterquality.org/bass/htm/bs_biomon.htm
8-9
-------
APPENDIX A: PROCESS AND CRITERIA FOR THE ASSIGNMENT OF BIOLOGIST'S
CLASSIFICATION
DRAFT
From: Appendix H in Stream Biological Monitoring and Numeric Criteria Development in
Maine
by:
Susan P. Davies, M.S.1
Leonidas Tsomides, M.S.1
David L. Courtemanch, Ph.D.1
Francis Drummond, Ph.D.2
1 Maine Department of Environmental Protection, Augusta, Maine
2 Department of Biological Sciences, University of Maine, Orono, Maine
Raters
David Courtemanch
MS in aquatic entomology; PhD in environmental science; employed as a Biologist in the
Division of Environmental Evaluation and Lake Studies (DEELS) in the Water Bureau for 16
years; currently Director, Division of Environmental Assessment.
Susan Davies
MS aquatic entomology; employed as a Biologist in the River and Stream section of
DEELS for 9 years, coordinating the Instream Biological Monitoring Program.
Leon Tsomides
MS aquatic entomology; employed as a Biologist in the River and Stream Section of
DEELS for 3 years, working with the Instream Biological Monitoring Program.
Ranking Process
Each biologist independently reviewed biological information for each sampling event, as
listed below, including identities and abundances of taxa occurring in the biological sample and
computed index values for the biological data (e.g. diversity, richness, EPT, etc). Physical habitat
information was also reviewed including water depth, velocity, substrate composition, canopy
cover, etc., in order to evaluate the effects of various habitat conditions on the structure of the
macroinvertebrate assemblage. Sample information was reviewed for the values of the given
measures, relative to values for other samples in the data set. The actual classification
assignment was determined by how closely the biological information conformed to the aquatic
life classification standards, correcting for habitat effects. Numerical ranges, per se, were not
established, a priori, for each measure. Instead, the information was reviewed for it's
compatibility with the mosaic of findings expected for each Class, listed in the Relative Findings
Chart in this Appendix (H-l). The biologists did not have any knowledge of the actual location
of the sampled sites, nor did they have knowledge of any pollution influences. Following the
A-l
-------
independent assignment of classes the biologists established a consensus classification, following
an open exchange of justifications for each biologist's assignment.
Biologist's Classification Criteria
Each biologist reviewed the sample data for the values of a list of measures of
community structure and function. Criteria used by biologists to evaluate each measure are listed
in the Relative Findings Chart, Appendix A-l.
TOTAL ABUNDANCE OF INDIVIDUALS
TOTAL ABUNDANCE OF EPHEMEROPTERA
TOTAL ABUNDANCE OF PLECOPTERA
ABUNDANCE OF EPHEMEROPTERA/TOTAL ABUNDANCE
ABUNDANCE OF PLECOPTERA/TOTAL ABUNDANCE
ABUNDANCE OF HYDROPSYCHIDAE/TOTAL ABUNDANCE
ABUNDANCE OF EPHEMEROPTERA+PLECOPTERA/TOTAL ABUNDANCE
ABUNDANCE OF GLOSSOSOMATTQTAL ABUNDANCE
ABUNDANCE OF BRACHYCENTRUSIrTQrTP^L ABUNDANCE
ABUNDANCE OF OLIGOCHAETES/TOTAL ABUNDANCE
ABUNDANCE OF HIRUDINEA/TOTAL ABUNDANCE
ABUNDANCE OF GASTROPODA/TOTAL ABUNDANCE
ABUNDANCE OF CHIRONOMIDAE/TOTAL ABUNDANCE
ABUNDANCE CONCHAPELOPIA + THIENNEMANNYMIAIWTAL ABUNDANCE
ABUNDANCE OF TRIBELOSITQTkL ABUNDANCE
ABUNDANCE OF CHIRONOMUSIrTQrTP^L ABUNDANCE
GENERIC RICHNESS
EPHEMEROPTERA RICHNESS
PLECOPTERA RICHNESS
EPT RICHNESS
EPHEMEROPTERA RICHNESS/GENERIC RICHNESS
PLECOPTERA RICHNESS/GENERIC RICHNESS
DIPTERA RICHNESS/GENERIC RICHNESS
EPHEMEROPTERA+PLECOPTERA RICHNESS/GENERIC RICHNESS
EPT RICHNESS/DIPTERA RICHNESS
NON-EPT OR CHIRONOMIDAE RICHNESS/GENERIC RICHNESS
PERCENT PREDATORS
% COLLECTOR FILTERERS+GATHERERS/%PREDATORS+SHREDDERS
NUMBER OF FUNCTIONAL FEEDING GROUPS REPRESENTED
SHANNON-WEINER GENERIC DIVERSITY
HILSENHOFF BIOTIC INDEX
In addition, in cases where a valid, clean-water, upstream reference station existed, the
following comparative index data was also reviewed:
JACCARD TAXONOMIC SIMILARITY
TAXONOMIC SIMILARITY OF DOMINANT TAXA
COEFFICIENT OF COMMUNITY LOSS
A-2
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PERCENT SIMILARITY
Results
In 64% of the cases there was unanimous agreement among the independent raters, and in
an additional 34% of the samples two of the raters were in agreement and one had assigned a
different classification. In 3 of the rated samples there was disagreement among all three raters
Table A-l. RELATIVE FINDINGS CHART
Measure of
Community
Structure
Total Abundance
of Individuals
Abundance of
Ephemeroptera
Abundance of
Plecoptera
Proportion of
Ephemeroptera
Proportion of
Pleocoptera
Proportion of
Hydropsychidae
Proportion of
Ephemeroptera &
Plecoptera
Proportion of
Glossoma
Proportion of
Brachycentrus
Proportion of
Oligochaetes
Proportion of
Hirudinea
Proportion of
Gastropoda
Proportion of
Chironomidae
Relative Findings
A
often low
high
highest
highest
highest
intermediate
highest
highest
highest
low
low
low
lowest
B
often high
high
some present
variable,
depending on
dominance by
other groups
variable,
depending on
dominance by
other groups
highest
variable
low to
intermediate
low to
intermediate
low
variable
low
variable,
depending on the
dominance of
other groups
C
variable
low
low to absent
low
low
variable
Low
very low to
absent
very low to
absent
low to moderate
variable
variable
highest
NA
variable: often
very low or high
low to absent
absent
zero
zero
low to high
absent
absent
absent
highest
variable to
highest
variable to
highest
variable
A-3
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Measure of
Community
Structure
Proportion of
Conchapelopia &
Thienemannimyia
Proportion of
Tribelos
Proportion of
Chironomus
Generic Richness
Ephemeroptera
Richness
Plecoptera
Richness
EPT Richness
Proportion
Ephemeroptera
Richness
Proportion
Plecoptera
Richness
Proportion
Diptera Richness
Proportion
Ephemeroptera &
Plecoptera
Richness
EPT Richness
dvided by Diptera
Richness
Proportion Non-
EPTor
Chronomid
Richness
Percent Predators
Percent Collector,
Filterers &
Gatherers divided
by Percent
Predators &
Shredders
Number of
Functional
Feeding Groups
Relative Findings
A
lowest
low to absent
low to absent
variable
highest
highest
high
highest
highest
low to variable
highest
high
high
low
high
variable
B
low to variable
low to absent
low to absent
highest
high
variable
highest
variable
high
variable
high
highest
high
low
highest
highest
C
variable
low to variable
low to variable
variable
low
low to absent
variable
low
low
highest
low to variable
low to variable
low
high to variable
low
variable
NA
variable to
highest
variable to
highest
variable to
highest
lowest
very low to
absent
absent
low
zero
low to zero
variable to high
low to absent
lowest to zero
lowest
highest
lowest
lowest
A-4
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Measure of
Community
Structure
Represented
Shannon-Weiner
Generic Diversity
Hilsenhoff Biotic
Index
Relative Findings
A
low to
intermediate
lowest
B
highest
low
C
variable to
intermediate
intermediate
NA
lowest
highest
A-5
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vvEPA
United States
Environmental Protection
Agency
Office of Research and Development
National Exposure Research Laboratory
Cincinnati, OH 45268
Official Business
Penalty for Private Use
$300
EPA/600/R-04/168
October 2004
PRESORTED STANDARD
POSTAGE & FEES PAID
EPA PERMIT NO. G-35
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