United States Environmental Office of Water EPA-822-R-03-013
Protection Agency Washington, DC 20460 May 2003
METHODS FOR EVALUATING WETLAND CONDITION
#14 Wetland Biological Assessment
Case Studies
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United States Environmental Office of Water EPA-822-R-03-013
Protection Agency Washington, DC 20460 May 2003
METHODS FOR EVALUATING WETLAND CONDITION
#14 Wetland Biological Assessment
Case Studies
Edited By
Maine Department of Environmental Protection
Thomas J. Danielson
Connecticut Department of Environmental Protection
Douglas G. Hoskins
Prepared jointly by:
The U.S. Environmental Protection Agency
Health and Ecological Criteria Division (Office of Science and Technology)
and
Wetlands Division (Office of Wetlands, Oceans, and Watersheds)
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NOTICE
The material in this document has been subjected to U.S. Environmental Protect on Agency (EPA)
technical review and has been approved for publication as an EPA document. The information
contained herein is offered to the reader as a review of the "state of the science" concerning wetland
bioassessment and nutrient enrichment and is not intended to be prescriptive guidance or firm advice.
Mention of trade names, products or services does not convey, and should not be interpreted as
conveying official EPA approval, endorsement, or recommendation.
APPROPRIATE CITATION
U.S. EPA. 2003. Methods for Evaluating Wetland Condition: Wetland Biological Assessment
Case Studies. Office of Water, U.S. Environmental Protection Agency, Washington, DC.
EPA-822-R-03-013.
ACKNOWLEDGMENTS
EPA acknowledges the contributions of the following people in the writing of this module: The members
of B AWWGwithout their hard work, this report would not have possible. Leigh Dunkelberger
provided invaluable assistance during her internship with the U. S. EPA Wetlands Division.
This entire document can be downloaded from the following U.S. EPA websites:
http://www.epa.gov/ost/standards
http://www.epa.gov/owow/wetlands/bawwg
11
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CONTENTS
FOREWORD vn
LIST OF "METHODS FOR EVALUATING WETLAND CONDITION" MODULES IX
FLORIDA: MONITORING ACROSS A NUTRIENT GRADIENT IN THE EVERGLADES 1
FLORIDA: DEVELOPMENT OF A BIOLOGICAL APPROACH FOR
ASSESSING FLORIDA WETLAND INTEGRITY 7
MAINE: DEVELOPING A STATEWIDE BIOLOGICAL MONITORING
AND ASSESSMENT PROGRAM FOR FRESHWATER WETLANDS 14
MARYLAND: DEVELOPING AN IBI ASSESSMENT FOR RESTORED
WETLANDS IN THE MID-ATLANTIC STATES 18
MASSACHUSETTS: USE OF MULTIMETRIC INDICES TO EXAMINE ECOLOGICAL
INTEGRITY OF SALT MARSH WETLANDS IN CAPE COD 22
MASSACHUSETTS: INVOLVING VOLUNTEERS IN EXAMINING THE
ECOLOGICAL INTEGRITY OF COASTAL WETLANDS IN CAPE COD,
MASSACHUSETTS 3O
MICHIGAN: BIOASSESSMENT PROCEDURES AND BASELINE REFERENCE
DATA FOR GREAT LAKES COASTAL MARSHES AND INLAND FORESTED
WETLANDS IN MICHIGAN 32
MINNESOTA: DEVELOPING WETLAND BIOCRITERIA 37
MINNESOTA: DAKOTA COUNTY WETLAND HEALTH EVALUATION PROJECT 44
MINNESOTA: UNIVERSITY OF MINNESOTA'S IBI DEVELOPMENT 47
MONTANA: DEVELOPING WETLAND BIOASSESSMENT PROTOCOLS To
SUPPORT AQUATIC LIFE BENEFICIAL USE-SUPPORT DETERMINATIONS 49
NORTH DAKOTA: WETLAND BIOASSESSMENT PROTOCOLS FOR MAKING
AQUATIC LIFE BENEFICIAL USE-SUPPORT DETERMINATIONS 59
OHIO: DEVELOPING IBI ASSESSMENT METHODS FOR WATER QUALITY
STANDARDS AND REGULATORY DECISIONS 63
OREGON: SIMULTANEOUS DEVELOPMENT, CALIBRATION, AND TESTING
OF HYDROGEOMORPHIC-BASED (HGM) ASSESSMENT PROCEDURES AND
BIOLOGICAL ASSESSMENT PROCEDURES 72
iii
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PENNSYLVANIA: ASSESSING WETLAND CONDITION AT A WATERSHED
SCALE USING HYDROGEOMORPHIC MODELS AND MEASURES OF
BIOLOGICAL INTEGRITY 75
VERMONT: CLASSIFICATION, BIOLOGICAL CHARACTERIZATION, AND
BIOMETRIC DEVELOPMENT FOR NORTHERN WHITE CEDAR SWAMPS
AND VERNAL POOLS 77
WASHINGTON: KING COUNTY WETLAND-BREEDING AMPHIBIAN
MONITORING PROGRAM 8O
WISCONSIN: DEVELOPING BIOLOGICAL INDEXES FOR WISCONSIN'S
PALUSTRINE WETLANDS 82
WISCONSIN: REFINEMENT AND EXPANSION OF THE WISCONSIN WETLAND
BIOLOGICAL INDEX FOR ASSESSMENT OF DEPRESSIONAL PALUSTRINE
WETLANDS 86
REFERENCES 91
GLOSSARY 93
LIST OF FIGURES
FIGURE l: AREA OF INTEREST WITHIN EVERGLADES PROTECTION AREA l
FIGURE 2: WCA 2A MONITORING SITES ALONG THE PHOSPHORUS GRADIENT .. 2
FIGURE 3: CHANGE POINT ANALYSES OF ELEOCHARIS FREQUENCY OF
OCCURRENCE AND BIOMASS DATA ALONG THE SFWMD
TRANSECTS 5
FIGURE 4: RESULTS OF CHANGE POINT ANALYSES PERFORMED ON
MEDIAN TOTAL PERCENTAGE OF POLLUTION-SENSITIVE
(LITERATURE DETERMINED) PERIPHYTON TAXA 5
FIGURE 5: RESULTS OF CHANGE POINT ANALYSES ON
MEDIAN FLORIDA INDEX VALUES 6
FIGURE 6: FLORIDA'S WETLAND REGIONS AND FIELD SITE LOCATIONS 9
FIGURE 7: RELATIONSHIPS BETWEEN WETLAND ECOLOGICAL CONDITION
AND LAND USE 27
FIGURE 8: Two SETS OF MULTIMETRIC GOALS FOR EVALUATING
RESTORATION 28
IV
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FIGURE 9: LOCATIONS OF SMALLER DEPRESSIONAL WETLANDS
SAMPLED IN 1995
.39
FIGURE 1O: LOCATIONS OF THE 44 LARGE DEPRESSIONAL WETLANDS
SAMPLED IN 1999
.39
FIGURE 11: MONTANA ECOREGIONS AND PILOT WATERSHEDS 51
FIGURE 12: ECOREGIONS AND SAMPLING LOCATIONS BY WETLAND CLASS 52
FIGURE 13: DIATOM DATA RELATED TO ABIOTIC FACTORS USING
DETRENDED CANONICAL CORRESPONDENCE ANALYSIS.
,54
FIGURE 14: CLUSTERS OF WETLANDS BASED ON DIATOM DATA 54
FIGURE 15: MACROINVERTEBRATE INDEX SCORES FOR WETLANDS
IN SEVERAL CLASSES
57
FIGURE 16: LEVEL IV ECOREGIONS OF NORTH DAKOTA 61
FIGURE 17: ECOLOGICAL REGIONS OF OHIO AND INDIANA 64
LIST OF PLATES
PLATE l: CYPRESS WETLAND 12
PLATE 2: MACROINVERTEBRATE SAMPLING 13
PLATE 3: MAINE'S STOVEPIPE SAMPLER AND SIEVE BUCKET 16
PLATE 4: PICKING MACROINVERTEBRATES FROM MULTIHABITAT SAMPLE 16
PLATE 5: CAPE COD, MA, SALT MARSH PLANT SURVEY.
PLATE 6: CAPE COD, MA, SALT MARSH NEKTON: WATCH YOUR FINGERS!
(BLUE CRAB, CALLINECTES SAPIDUS)
PLATE 7: IMPAIRED WETLAND 66
PLATE 8: LEAST-DISTURBED WETLAND 66
v
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LIST OF TABLES
TABLE l. SCORING CRITERIA FOR 1O INVERTEBRATE METRICS FOR IBI
FOR LARGE DEPRESSIONAL WETLANDS 42
TABLE 2. SCORING CRITERIA FOR TO VEGETATION METRICS FOR LARGE
DEPRESSIONAL WETLANDS 43
TABLE 3: PROPOSED METRICS, PROPOSED METRIC CALCULATIONS,
AND SCORE CALCULATIONS USED FOR DEVELOPING WETLAND
MACROINVERTEBRATE INDICES 56
VI
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FOREWORD
In 1999, the U.S. Environmental Protection Agency (EPA) began work on this series of reports entitled
Methods for Evaluating Wetland Condition. The purpose of these reports is to help States and
Tribes develop methods to evaluate (1) the overall ecological condition of wetlands using biological
assessments and (2) nutrient enrichment of wetlands, which is one of the primary stressors damaging
wetlands in many parts of the country. This information is intended to serve as a starting point for States
and Tribes to eventually establish biological and nutrient water quality criteria specifically refined for
wetland waterbodies.
This purpose was to be accomplished by providing a series of "state of the science" modules concerning
wetland bioassessment as well as the nutrient enrichment of wetlands. The individual module format
was used instead of one large publication to facilitate the addition of other reports as wetland science
progresses and wetlands are further incorporated into water quality programs. Also, this modular
approach allows EPA to revise reports without having to reprint them all. Alist of the inaugural set of 20
modules can be found at the end of this section.
This series of reports is the product of a collaborative effort between EPAs Health and Ecological
Criteria Division of the Office of Science and Technology (OST) and the Wetlands Division of the
Office of Wetlands, Oceans and Watersheds (OWOW). The reports were initiated with the support
and oversight of Thomas J. Danielson (OWOW), Amanda K. Parker and Susan K. Jackson (OST),
and seen to completion by Douglas G. Hoskins (OWOW) and Ifeyinwa F. Davis (OST). EPA relied
heavily on the input, recommendations, and energy of three panels of experts, which unfortunately have
too many members to list individually:
Biological Assessment of Wetlands Workgroup
New England Biological Assessment of Wetlands Workgroup
Wetlands Nutrient Criteria Workgroup
More information about biological and nutrient criteria is available at the following EPA website:
http://www.epa.gov/ost/standards
More information about wetland biological assessments is available at the following EPA website:
http://www.epa.gov/owow/wetlands/bawwg
vn
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LIST OF "METHODS FOR EVALUATING WETLAND
CONDITION" MODULES
MODULE # MODULE TITLE
1 INTRODUCTION TO WETLAND BIOLOGICAL ASSESSMENT
2 INTRODUCTION TO WETLAND NUTRIENT ASSESSMENT
3 THE STATE OF WETLAND SCIENCE
4 STUDY DESIGN FOR MONITORING WETLANDS
5 ADMINISTRATIVE FRAMEWORK FOR THE IMPLEMENTATION OF A
WETLAND BIOASSESSMENT PROGRAM
6 DEVELOPING METRICS AND INDEXES OF BIOLOGICAL INTEGRITY
7 WETLANDS CLASSIFICATION
8 VOLUNTEERS AND WETLAND BIOMONITORING
9 DEVELOPING AN INVERTEBRATE INDEX OF BIOLOGICAL
INTEGRITY FOR WETLANDS
10 USING VEGETATION TO ASSESS ENVIRONMENTAL CONDITIONS
IN WETLANDS
11 USING ALGAE TO ASSESS ENVIRONMENTAL CONDITIONS IN
WETLANDS
12 USING AMPHIBIANS IN BlOASSESSMENTS OF WETLANDS
13 BIOLOGICAL ASSESSMENT METHODS FOR BIRDS
14 WETLAND BIOASSESSMENT CASE STUDIES
15 BIOASSESSMENT METHODS FOR FISH
16 VEGETATION-BASED INDICATORS OF WETLAND NUTRIENT
ENRICHMENT
17 LAND-USE CHARACTERIZATION FOR NUTRIENT AND SEDIMENT
RISK ASSESSMENT
18 BIOGEOCHEMICAL INDICATORS
19 NUTRIENT LOAD ESTIMATION
2O SUSTAINABLE NUTRIENT LOADING
IX
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FLORIDA: MONITORING ACROSS A NUTRIENT
GRADIENT IN THE EVERGLADES
Contact
Russel Frydenborg
Organization Florida Department of Environmental Protection
2600 Blair Stone Road, MS 6511
Tallahassee, FL 32399-2400
Phone (850)921-9821
E-mail oissel.frydenborg@dep.state.fl.us
PURPOSE
rhis proj ect was initiated to monitor biologi-
cal assemblages across a nutrient gradient in
the Florida Everglades in support of regulatory ef-
forts to define a numeric water quality criterion for
phosphorus. The goal is protection of natural popu-
lations of aquatic flora and fauna in the Everglades
Protect on Area.
WETLAND TYPE
Freshwater marshes
ASSEMBLAGES
Algae
Macroinvertebrates
Vascular plants
STATUS
Revising sampling methods and analyzing data
PROJECT DESCRIPTION
The historic Florida Everglades consisted of ap-
proximately 4 million acres of shallow sawgrass
marsh, with wet prairies and aquatic sloughs inter-
spersed with tree islands. Today, only 50% of the
original Everglades ecosystem remains, primarily as
a result of drainage and conversion of large por-
tions of the northern and eastern Everglades to ag-
ricultural or urban land use. The remaining portions
of the historic Everglades are located in the Water
Conservation Areas (WCAs) and Everglades Na-
tional Park (ENP) (see Figure 1).
Sftndta
-^ -
-
Area oflnlcrvsl within
\alludes Protection Area
. . ,
FIGURE l: AREA OF INTEREST WITHIN
EVERGLADES PROTECTION AREA.
The Everglades ecosystem evolved under ex-
tremely low phosphorus concentrations and is con-
sidered an oligotrophic ecosystem. Alargebody
of evidence indicates that phosphorus is the pri-
mary limiting nutrient throughout the remaining
Everglades. Introduction of excess phosphorus to
the Everglades has resulted in ecological changes
over large areas of the marsh. The Everglades For-
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ever Act (EFA; Section 373.4592, Florida Stat-
utes), passed by the Florida Legislature in 1994,
stated that waters flowing into the part of the rem-
nant Everglades known as the Everglades Protec-
tion Area (defined as Water Conservation Areas 1,
2A, 2B, 3 A, 3B, and ENP) contain excessive lev-
els of phosphorus and that a reduction in levels of
phosphorus will benefit the ecology of the Ever-
glades Protection Area. The EFA requires the
Florida Department of Environmental Protection
(FDEP) and the South Florida Water Management
District (SFWMD) to complete research necessary
to establish a numeric phosphorus criterion for the
Everglades Protection Area.
The SFWMD Everglades System Research Di-
vision (ESRD) initiated a succession of studies, be-
ginning in 1993 and continuing to the present, as
part of the research and monitoring being conducted
in the Everglades for phosphorus criterion devel-
opment. Biological monitoring for the ESRD stud-
ies was initiated in early 1994 in WCA 2A. Data
from this and other studies are being used by FDEP
in the development of a numeric phosphorus crite-
rion for the Everglades Protection Area.
STUDY DESIGN
SFWMD ESRD initially selected 13 sites along 2
transects located downstream of canals discharg-
ing into WC A 2A and extending down the phos-
phorus gradient into the least affected areas of the
marsh. Sampling sites ranged from the canal in-
flows (discharge structures on the northeastern
margin of WC A 2 A) to a site nearly 15 km down-
stream from the canal inflows. Three of the 13 main
sites (sites U1-U3) were specifically chosen to rep-
resent the least affected area of WC A 2A with re-
spect to anthropogenic disturbance. A series of 15
additional "intermediate" sites were added to the
study later to obtain better spatial coverage of the
lower portion of the transects. The sites have been
monitored for water, sediment, and biological qual-
ity. Figure 2 shows the WCA2Amonitoring sites
located along the phosphorus gradient.
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FIGURE 2: WCA 2A MONITORING
SITES ALONG THE PHOSPHORUS
GRADIENT.
SAMPLING METHODS: ALGAE
Water Bottles: Phytoplankton samples initially
were collected monthly and later collected
quarterly using water bottles. Samples were
preserved in the field and sent to the FDEP
Central Biology Laboratory for taxonomic iden-
tification.
Diatometers: Racks each containing six glass
diatometer slides were deployed quarterly at
each site. It was determined that an 8-week
deployment was necessary for sufficient per-
iphyton growth. Diatometers were collected
and preserved and sent to the FDEP Central
Biology Laboratory for processing and taxo-
nomic identification.
Natural Substrate (benthic): Samples of
benthic periphyton were collected from surficial
sediment cores at the main transect sites on sev-
eral occasions. Samples were retained by
SFWMD ESRD for processing and taxonomic
identification.
ANALYTICAL METHODS: ALGAE
Water Bottles: Samples were processed and
enumerated by FDEP Central Biology Labo-
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ratory staff according to FDEP standard oper-
ating procedures (SOPs) (e.g., AB-04 and AB-
05; available at http://www8.myflorida.com/
labs/sop/index.htm). Analysis from this study
and other studies has indicated that Everglades
phytoplankton are largely periphyton that has
sloughed off into the water column. Thus, algal
data analysis was focused on the periphyton
data.
Diatometers: Samples were processed and
enumerated by FDEP Central Biology Labo-
ratory staff according to FDEP SOPs (e.g.,
AB-02, AB-02.1, AB-02.2, andAB-03; avail-
able at http://www8.myflorida.com/labs/sop/
index.htm).
Natural Substrate (benthic): SFWMD pro-
cessed and enumerated natural substrate
samples.
SAMPLING METHODS:
MACROINVERTEBRATES
Dipnet: SFWMD staff conducted quarterly
macroinvertebrate sampling using a standard D-
frame dipnet with a 30-mesh bag from Sep-
tember 1994 through November 1995. The
sampling method consisted of the collection of
20 0.5 m (in length) discrete dipnet sweeps from
representative habitats in the area of each site
on a given sampling date. The 20 dipnet sweeps
for a given site were combined and sent to the
FDEP Central Biology Laboratory for process-
ing and taxonomic identification.
QuanNet: Beginning in May 1996, SFWMD
staff conducted quarterly macroinvertebrate
sampling using the Quan Net method. The sam-
pling method consisted of the deployment of a
1 -m2 frame at the site and the collection of net
samples and all vegetation within the area of
the frame. Frames were deployed in each of
several representative habitats, where present,
in the vicinity of each site. Samples from each
site/habitat were kept separate. Representa-
tive habitats were labeled as cattail, sawgrass,
or slough, depending on the predominant veg-
etation type. The collected material from each
site/habitat was subsampled, preserved, and
sent to the FDEP Central Biology Laboratory
for processing and taxonomic identification.
Hester-Dendy: SFWMD staff deployed
Hester-Dendy artificial substrate samplers at
each of the main transect sites on a quarterly
basis. The samplers were deployed for a 1-
month period, after which they were collected,
preserved, and sent to the FDEP Central Biol-
ogy Laboratory for processing and taxonomic
identification.
ANALYTICAL METHODS:
MACROINVERTEBRATES
Dipnet and Quan Net: FDEP Central Biol-
ogy Laboratory staff subsampled the dipnet
and Quan Net samples from each site and anal-
yzed them according to FDEP SOPs
(e.g., IZ-02 and IZ-06; available at
http://www8.myflorida.com/labs/sop/
index.htm).
Hester Dendy: FDEP Central Biology Labo-
ratory staff processed and analyzed the Hester-
Dendy samples from each site according to
FDEP SOPs (e.g., IZ-03 and IZ-06; available
at http://www8.myflorida.com/labs/sop/
index.htm).
SAMPLING METHODS: MACROPHYTES
Macrophyte Stem Density andFrequency: In
April 1997, SFWMD staff conducted a study
of macrophytes at the WC A 2A transect sites.
A 50-m tape was laid out at each transect site.
A1 -m square frame was used every 2 m along
the tape to delineate the sample area for calcu-
lation of macrophyte stem densities (stems/m2)
and frequencies (# plots where a species was
found/total # of plots) by species.
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Macrophyte Harvesting: On the other side
of the 50-m tape used for establishing stem den-
sities and frequencies, SFWMD staff harvested
macrophytes for biomass measurements, using
the 1 -m square frame at five predetermined lo-
cations to mark the sample area for harvesting.
ANALYTICAL METHODS:
MACROPHYTES
Macrophyte Stem Density and Frequency:
Stem densities (stems/m2) and frequencies (#
plots where a species was found/total # of plots)
by species were counted at each site.
Macrophyte Harvesting: SFWMD staff con-
ducted biomass analysis of the harvested mac-
rophytes for comparison of the relative biom-
ass of several species present at each of the
WCA 2A transect sites (e.g., Eleocharis,
Nymphaea, Typha).
LESSONS LEARNED
Periphyton, macroinvertebrate, and macrophyte
communities in WCA 2A change sub stantially from
reference conditions at approximately 7 to 8 km
downstream of canal discharges into WCA 2 A (see
Figures 3-5). Data analysis has shown that bio-
logical populations at the two stations (E5 and F5)
nearest to the three initial reference sites (Ul -U3)
are very similar in terms of biological community
structure. This suggests that these areas, despite
slight phosphorus enrichment, still support reference
condition biota. The somewhat higher phosphorus
regimes at the next stations (E4 and F4 and be-
yond) are associated with greater biological changes.
Experimental field dosing studies (microcosms) have
been conducted by SFWMD ESRD and show that
the addition of phosphorus causes changes in per-
iphyton assemblages consistent with those observed
in the transect study.
The WCA 2A transect periphyton data for each
site/date have been analyzed using the entire taxo-
nomic assemblage encountered and using lists of
pollution-sensitive and -tolerant species based on
available literature and experimental phosphorus
addition studies (the microcosms) in WCA 2A.
Macroinvertebrate data have been analyzed using
the Florida Index and the macroinvertebrate com-
ponent of the Lake Condition Index (LCI), mea-
sures of the numbers of pollution-sensitive taxa in a
sample that are routinely used by FDEP in
bioassessments of streams and lakes. The use of
these methods with the WCA 2A transect data has
demonstrated a clear signal of biological disturbance
along the nutrient gradient in WCA 2A. FDEP is
using this information as well as information from
other studies conducted in the Florida Everglades
to develop a numeric phosphorus criterion for the
Everglades Protection Area.
ADDITIONAL COMMENTS
The information provided here is based solely on
the transect study by SFWMD ESRD in WCA 2A.
Research and monitoring of Florida Everglades
water, sediment and biological quality is being con-
ducted by several research groups in WCA 2A,
WCA 1 (Arthur R. Marshall Loxahatchee National
Wildlife Refuge), Everglades National Park (ENP),
and WCA 3 A, including studies by SFWMD ESRD
similar to the WCA 2Atransect study.
4
-------
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6 8 10 12
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FIGURE 3: CHANGE POINT ANALYSES OF ELEOCHARIS FREQUENCY OF
OCCURRENCE AND BIOMASS DATA ALONG THE SFWMD TRANSECTS.
COLLECTED APRIL 1997.
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FIGURE 4: RESULTS OF CHANGE POINT ANALYSES PERFORMED ON
MEDIAN TOTAL PERCENTAGE OF POLLUTION-SENSITIVE
(LITERATURE DETERMINED) PERIPHYTON TAXA.
-------
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Contact
FLORIDA: DEVELOPMENT OF A BIOLOGICAL
APPROACH FOR ASSESSING FLORIDA
WETLAND INTEGRITY
Mark Brown
Organization University of Florida, Center for Wetlands
PhelpsLab-POBoxll6350
Gainesville,FL 32611-6350
Phone (352) 392-2309
E-mail mtb@ufl.edu
Website http://www.enveng.ufl.edu/homepp/brown/syseco
PURPOSE
r I To develop "bioindicators" of ecosystem
J. health for wetlands in Florida. To achieve this
goal, the project team has developed wetland
ecoregions using GIS technology, a classification
scheme for Florida wetlands, sampling protocols
for herbaceous and forested wetlands, and a quan-
titative index of the human disturbance gradient.
WETLAND TYPE
Freshwater marshes
Forested wetlands
ASSEMBLAGES
Algae
Macroinvertebrates
Vascular plants
STATUS
Developed field protocols, sampled more than 150
wetlands statewide, developed candidate plant,
macroinvertebrate, and algae metrics for marshes.
Currently sampling and developing metrics for for-
ested wetlands.
PROJECT DESCRIPTION
The University of Florida Center for Wetlands is
involved in a multiyear wetland research project
funded by the Florida Department of Environmen-
tal Protection (FDEP) to develop an integrated bio-
logical approach for evaluating Florida's wetlands.
The project goal is to develop an assessment ap-
proach that recognizes the utility of both biological
and functional assessments, and is rapid, reproduc-
ible, and meaningful.
The Center for Wetlands began development of
the assessment approach in 1997. Now in its fifth
year, the project, titled "Development of aBiologi-
7
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cal Approach for Assessing Wetland Function and
Integrity," has four main tasks:
Task 1. Review and synthesize all current and
relevant literature.
Task 2. Develop a wetland classification system
for wetland types in the State of Florida.
Task 3. Develop a GIS-driven methodology for
classifying bioregions within the State of Florida that
identify climatic, geologic, and geophysical prov-
inces that are sensitive to wetland classes.
Task 4. Develop a bioassessment methodology,
biocriteria, and metrics for wetlands in the State of
Florida.
To date, Tasks 1-3 have been completed, and
work continues on Task 4. Nearly complete is a
set of metrics for herbaceous depressional wetlands,
and data are currently being analyzed to develop
metrics for forested depressional wetlands.
The development of the approach has included a
review of technical and scientific literature, a wet-
land classification and crosswalk, wetland
regionalization of Florida, and two wetland biologi-
cal surveys in summer 1998 and summer 1999. The
biological surveys were designed to test the validity
(and necessity) of the proposed wetland regions,
to identify the appropriate biological indicator taxa,
and to quantify the gradient of human disturbance.
STUDY DESIGN
The approach has included a review of technical
and scientific literature, development of a wetland
classification system and crosswalk with other clas-
sifications, wetland regionalization of Florida, and
4 years of wetland biological surveys in the grow-
ing seasons of 1998,1999,2000, and 2001 (a fifth
season of field surveys is underway for 2002). The
biological surveys were designed to test the validity
(and necessity) of the proposed wetland regions,
to identify the appropriate biological indicator taxa,
and to quantify the gradient of human disturbance.
The wetland classification scheme was organized
with major classes defined from three variables: (1)
dominant vegetation (forested, shrub, herbaceous);
(2) geomorphic position (stream channel [flood-
plain], flat topography, sloped topography, lake
fringe, depressional); and (3) primary water source
(rainfall, surface water, ground water). Subclasses
are discriminated by modifiers (hydroperiod and
plant community associations). Eleven wetland
classes were identified: forested (river swamp,
slough/strand/seepage swamp, lake swamp, depres-
sion swamp, flatland swamp); shrub dominated
(shrub-scrub swamp); and herbaceous (river marsh,
wet prairie, lake marsh, depressional marsh, seep-
age marsh). An HTML electronic database has been
completed that crosswalks existing wetland classi-
fications to the new simplified classification scheme
developed for the bioassessment proj ect.
Regionalization of the State was necessary be-
cause there is significant variation in climatic and
physical features of the Florida peninsula and it was
believed that these regional differences would equate
to variations in bioindicator "signals." Map cover-
ages of physical and climatic variables of the Florida
landscape were used to develop regions that had
different characteristic driving energies and land-
scape structural characteristics.
The map coverages were combined with GIS map
algebra to create a spatial hydrologic budget equa-
tion for the State. The equation modeled the move-
ment of water on the landscape during the ecologi-
cally sensitive spring growing season. The output
of the model provided a value for a Potential Soil
Moisture Index (PSMI). The PSMI was separated
into four regions based on both the critical depth of
saturation and on a statistical clustering of the PSMI
values (see Figure 6). The classified regions were
tested for similarities and differences to determine if
between-region variation in wetland type and envi-
ronmental variables was greater than within-region
variations.
8
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1HI South Region
I ] Central Region
BH North Region
I I Panhandle
A Forested Wetlands (72 sites)
O Marshes (77 sites)
FIGURE 6: FLORIDA'S WETLAND REGIONS AND FIELD SITE LOCATIONS.
The importance of hydrology in determining wet-
land type and location (the premise behind the
PSMI) was then tested using a hierarchical classifi-
cation technique (TWINSPAN) and ordination
(DCA and CCA) with variables of seasonal and
annual rainfall, seasonal and annual potential evapo-
transpiration, slope, geology, drainage class, and
runoff. TWINSPAN, DCA, and CCA tested the
relationships between wetland type and climatic and
physiographic landscape characteristics. Based on
the geostatistical output, hydrology is indeed a ma-
jor determinant of wetland type and location, and
supports the use of the spatial hydrologic budget
equation in delineating wetland regions.
Four years of biological surveys of wetlands
throughout the State were designed to test the
regionalization techniques as well as the metric and
bioindicator development. The 1998 pilot field re-
search involved surveying 24 herbaceous and for-
ested depressional wetlands in north and central
Florida. Maj or taxonomic assemblages were char-
acterized and ranked along a gradient of distur-
bances. Sites were located within multiple land uses
such as parks, preserves, pastures, farm fields, well
fields, silviculture plots, and urban areas. Impacts
that were assessed included hydrological modifica-
tions, nutrient loading, and altered hydroperiod. The
first year of sampling resulted in development of
standardized sampling procedures, design and
implementation of a statewide sampling program,
and identification of community attributes and can-
didate metrics.
In the second and third field seasons, 77 herba-
ceous, depressional wetlands were surveyed in 3
regions (south, central, and north). Approximately
half of the wetlands were impacted (agricultural set-
ting) and half were reference locations. Many of
the sites were paired sites (impacted and reference
at close proximity).
9
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In the fourth season, 72 forested, depressional
wetlands were surveyed in four regions (18 sites in
each of four regions: north, central, and southern
peninsular as well as the panhandle; see Figure 6).
Approximately ? of the wetlands were impacted (?
in agricultural settings and ? in urban settings) and ?
were reference locations. Many of the sites were
paired sites (impacted and reference at close prox-
imity).
Quantifying gradients of human disturbance
A Land Development Intensity (LDI) index is be-
ing used to quantify gradients of human disturbance
for wetlands throughout the State. The LDI index
is calculated using land use/land cover characteris-
tics, from aerial photographs, of lands within a 100-
m buffer surrounding the wetland. Land uses in the
area surrounding a wetland are first characterized
and then an intensity factor is assigned to each land
use type. The LDI algorithm multiplies the percent
area of each land use/land cover in the surrounding
100-m buffer by intensity coefficients. The inten-
sity coefficients are scaled from 1 to 10 and repre-
sent intensity of environmental manipulation as mea-
sured by energy use per unit area per unit time. The
LDI index is such that lower LDI values are indica-
tive of a lower disturbance level.
SAMPLING METHODS: ALGAE
Materials
Three 100-mL collection j ars
1-L collection jar
Bottomless collection cup, knife, large pipette,
bag, brush/scraper
M3 preservative
1 L of deionized water (for dry sites)
Methods
1. Methods vary depending on whether there is
water present at the site. We are experimen-
tally collecting dry benthic algae samples when
wetlands are dry. At this time, there is no infor-
mation as to the usefulness of this sampling tech-
nique. Methods outlined below are for wet
sites.
2. Samples are taken depending on substrate and
separated as benthic, epiphyton, metaphyton,
and phytoplankton.
3. For epiphyton, the 10 aliquots are divided
equally among herbaceous and woody debris:
a. For herbaceous debris, plant stems are cut
from the soil to the water surface, and
placed in a zip-lock bag with some wet-
land water. The plant matter is shaken and
massaged thoroughly to dislodge the al-
gae. Using a large pipette, lOmL of sample
are extracted into collection jar.
b. For woody debris, a brush is used to dis-
lodge algae. If the debris is small enough,
it is placed in a bag, similar to the herba-
ceous methods above. If the debris is
larger, a bottomless collection cup is used
to confine the sample, and it is brushed
while under water.
4. For benthic algae, a bottomless collection cup
is used to isolate a spot on the sediment. Then
a large pipette is used to gently stir the surface
(top 1 cm) of the sediment, and extract a 10-
mL aliquot that is placed in a collection jar.
Sampling is repeated at different locations until
a total of 100 mL is collected.
5. For metaphyton, a thumbnail size portion of the
algal mat is collected from 10 different loca-
tions throughout the wetland.
6. For phytoplankton, a total 100 mL of surface
water is collected at 10 locations throughout
the wetland.
7. All samples are preserved with M3 using 5 mL
per 100 mL of sample.
8. Samples are sent for later laboratory identifica-
tion.
10
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SAMPLING METHODS:
MACROINVERTEBRATES
Materials
Field Physical/Chemical Characterization Data
Sheet
Habitat Assessment Sheet
D-Frame dipnet with no. 30 mesh
4-L sample jars
Buffered formalin
Methods
1. The wetland is visually examined by walking
throughout the wetland, paying close attention
to its physical and habitat characteristics, and a
Field Physical/Chemical Characterization Data
Sheet and Habitat Assessment Sheet is com-
pleted. The percent coverage of substrate type
and each habitat type is recorded.
2. A total of 20 sweeps with a D-frame dipnet are
divided between the habitat types based on their
percentage of total wetland area. Discrete 0.5-
m sweeps are performed with the dipnet. The
available substrates are sampled as determined
by the above procedures in the following
manner:
a. For areas without flow, an area of sub-
strate that is one dipnet width wide and
approximately 0.5-m long is disturbed by
sweeping the net over the area three times
to ensure the capture of organisms.
b. For heavily vegetated areas, the net is
jabbed into the base of the vegetation, dig-
ging down to (but not into) the substrate,
and dislodging organisms using a 0.5-m
sweeping motion with the net.
c. Sand, muck, mud, and silt (nonmaj or habi-
tats) are sampled by taking 0.5-m sweeps
with the net while digging into the bottom
approximately 1 cm.
3. The numb er of sweep s for each habitat i s re-
corded on the Field Physical/Chemical Char-
acterization Data Sheet
4. The collected samples are reduced in volume
after each discrete sample by dislodging organ-
isms from larger debris (but retaining inverte-
brates in the net or sieve) and discarding the
debris. The finer debris and organisms are saved
in sample jars.
5. Samples are preserved with buffered formalin
and shipped to the FDEP Laboratory for iden-
tification.
SAMPLING METHODS: PLANTS
Materials
Field data sheets
Compass
100m of tape
FDEP's Florida Wetland Plant Identification
Manual (Tobeetal. 1998)
Methods
1. Using a compass (or the GPS unit) four line-
transects are located from four cardinal point
directions (N, S, E, W) that run parallel to the
slope of the wetland, beginning at the upland
edge (0 m) and extending into the center of the
wetland.
2. At the beginning of each transect, the approxi-
mate wetland/upland edge is located using a
combination of wetland plants and hydric soils.
3. Preferably, all four transects are set at one time,
with N, S and E, W transects meeting perpen-
dicularly in the center, and dividing wetland into
four equal sections. Each transect is started
with the 0-m point at the wetland-upland edge
and increasing in distance toward the wetland
interior. Aminimum of four data sheets is needed
per site.
1 1
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4. Two team members walk along each transect
and record species presence of all plants within
0.5 m on either side of each 5-m interval (sam-
pling area = 5 m2).
5. Plant species names are recorded on the data
sheets with full genus and species if known.
6. If unknown, the species is given aunique code
identifying transect location and number of un-
known encountered (N-l, N-2, N-3, E-l, E-
2, etc.). Voucher specimens for all unknown
species are collected, making sure to include
plant inflorescence and roots. Each specimen
is tagged with properly labeled masking tape,
and put in collection bag then pressed for later
laboratory identification.
7. Plant nomenclature follows FDEP's Florida
Wetland Plant Identification Manual (Tobe et
al. 1998).
SAMPLING METHODS:
WATER QUALITY AND SOILS
Water quality samples are collected, preserved,
and immediately sent to the chemistry laboratory
for analysis. Acomposite soil sample is taken from
each vegetation zone and later analyzed for nutri-
ents, organic matter content, and physical proper-
ties.
ANALYTICAL METHODS: PLANTS
Field data are entered into an MS Access data-
base for analysis. After entry, each data sheet is
checked by a second technician.
PLATE l: CYPRESS WETLAND. PHOTO: ELIZABETH SPURRIER
12
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PLATE 2: MACROINVERTEBRATE SAMPLING. PHOTO: KELLY CHINNERS REISS
13
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MAINE: DEVELOPING A STATEWIDE BIOLOGICAL
MONITORING AND ASSESSMENT PROGRAM
FOR FRESHWATER WETLANDS
Contact
Jeanne DiFranco
Organization Maine Department of Environmental Protection
312CancoRoad
Portland, ME 04103
Phone (207) 822-6424
E-mail Jeanne.L.Difranco@state.me.us
Website http ://www. state.me.us/dep/blwq/monitoring.htm#programs
PURPOSE
Examine regional differences in wetland
macroinvertebrate and algal assemblages
Test and refine candidate biological metrics
Diagnose stressors and identify risks to wet-
lands from human activities
Develop impairment thresholds andbiocriteria
for use in State water quality standards
WETLAND TYPE
Freshwater marshes
ASSEMBLAGES
Macroinvertebrates
Algae
STATUS
Expanding to Statewide program on a 5-year ro-
tating basin schedule, testing and refining metrics,
analyzing data, and developing impairment thresh-
olds.
PROJECT DESCRIPTION
In 1998, the Maine Department of Environmental
Protection (DEP) began development of a biologi-
cal monitoring and assessment program for fresh-
water wetlands. Between 1998 and 2000, DEP
conducted a pilot study in the Casco Bay water-
shed to develop wetland biomonitoring protocols
and identify candidate metrics related to wetland
condition. During 2001 and 2002, DEP expanded
monitoring to the Saco, Piscataqua, and Kennebec
River watersheds using the methods developed in
the pilot study. DEP uses aquatic macro-inverte-
brates as the primary taxonomic assemblage for thi s
program. Algae and diatoms are also collected as
part of a collaborative pilot proj ect undertaken by
Dr. R. Jan Stevenson of Michigan State University
to develop algal indicators of wetland integrity.
Based on the results of Dr. Stevenson's work, Maine
DEP will evaluate the usefulness of algae to its wet-
land biomonitoring program. Assessment of algae
has not been formally integrated into the Maine pro-
gram at this time, however.
14
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STUDY DESIGN
The Maine wetland biomonitoring initiative has
been incorporated into DEP's existing Biological
Monitoring Program, and will be extended to as-
sess additional major watersheds Statewide. Wet-
land monitoring is currently coordinated with the
State's river and stream program using the follow-
ing 5-year rotating basin schedule:
Kennebec, midcoast watersheds 2002
Androscoggin watershed 2003
St. John, Presumpscot watersheds 2004
Saco, southern coastal watersheds 2005
Penobscot, downeast watersheds 2006
Considerations for site selection include hydro-
logic regime, geographic distribution of sites, land-
scape position, human disturbance gradient, man-
agement significance, and accessibility. All wetlands
sampled are semipermanently or permanently inun-
dated, and range from minimally disturbed poten-
tial reference sites to poor-quality wetlands. As of
2002, DEP has conducted wetland biomonitoring
at 88 different sites encompassing 115 sampling
events. Some sites have been sampled repeatedly
over multiple years.
SAMPLING METHODS:
MACROINVERTEBRATES
Macroinvertebrates are currently sampled during
June and early July. Three different approaches have
been tested to develop both qualitative and quanti-
tative methods. In addition, water samples are ana-
lyzed for a suite of physical and chemical param-
eters to help in wetland characterization, and to iden-
tify potential sources of human impact. Habitat in-
formation, dominant plant species, and a scoring of
human disturbances are recorded in the field, along
with measurements of water temperature, pH, dis-
solved oxygen, and conductivity.
Multihabitat sampling
A qualitative, multihabitat sampling approach was
tested, with the goal of developing a screening level
assessment tool. A standard D-frame net was used
to sample all inundated microhabitats at each site,
including emergent vegetation, aquatic macrophyte
beds, pools, and channels. Samples were "picked"
or sorted from detritus in the field. One to several
organisms representing each different taxon found
were placed into a vial of alcohol until no different
taxa were observed.
Stovepipe sampler
Maine DEP designed its own stovepipe sampler
for quantitative samples usingaS-gallon bucket with
the bottom removed. In this method, the sampler
was used to enclose three replicate plots to restrict
the movement of organisms. The stovepipe sam-
pler was pressed into the wetland substrate, and
the contents of the sampler were then agitated.
Vegetation and surface sediment were placed into
a sieve bucket. The sampler was then swept 10
times with a small hand net. Large pieces of veg-
etation were washed and discarded; however, finer
plant material and detritus were retained. Samples
were preserved for later sorting and taxonomic
analysis in the laboratory.
Dipnet measured sweep
A standard D-frame net is currently used to ob-
tain a semi quantitative sample. A sample is col-
lected by submersing the net and sweeping through
the water column for a distance of 1 meter. The net
is bumped against the bottom substrate three times
(at the beginning, middle, and end of the sweep) to
dislodge and collect organisms from the sediment.
All material collected is placed in a sieve bucket.
Large pieces of vegetation are washed and dis-
carded; however, finer plant material and detritus
are retained. Three replicate samples are collected
in areas of emergent vegetation. Samples are pre-
served for later sorting and taxonomic analysis in
the laboratory.
15
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PLATE 3: MAINE'S STOVEPIP
SAMPLER AND SIEVE BUCKET.
PHOTO: MAINE DEP
PLATE 4: PICKING
MACROINVERTEBRATES FROM
MULTIHABITAT SAMPLE.
PHOTO: MAINE DEP
ANALYTICAL METHODS:
MACROINVERTEBRATES
Analyses performed to date reveal significant re-
lationships between a number of candidate inverte-
brate metrics and watershed development. Many
invertebrate metrics tested also appear to respond
to changes in water quality typically associated with
urban nonpoint source pollution, including elevated
conductivity and concentrations of anions, cations,
and nutrients. As new data are collected, candi-
date metrics will be tested and refined, and regional
differences and ecological linkages among wetlands
and other waters will be examined. DEP is also
developing impairment thresholds for wetlands.
This is a necessary first step to enable the State to
use biological monitoring data in wetland manage-
ment decisions and development of wetland-
specific water quality standards (designated uses
and biological criteria), and to assess wetland con-
dition and attainment status.
SAMPLING METHODS: ALGAE
Quantitative and qualitative algae samples were
gathered from the same wetland sites as used for
macroinvertebrate sampling. Four algae sample
types were tested to determine which produced the
best indicators. Samples were collected from the
water column, plant stems, and sediments. Samples
from multiple sites within each wetland were
composited into one sample from each habitat. In
addition, a multihabitat sample was collected from
each site. Samples were examined microscopically
to determine species numbers and relative abun-
dances of different species in samples. Chlorophyll
a was quantified from a separate water column
sample as an indicator of algal biomass.
For sampling, garden shears were used to clip
plant stems below the water line. Aturkey baster
was used to collect qualitative sediment samples;
however, this method was revised in 2002 to ob-
tain a more quantitative sample. Sediment is cur-
rently collected over a known surface area using a
petri dish pressed into the substrate and retrieved
with a spatula. Three replicates are collected and
composited into a single sediment sample. Along
handled dipper is used to collect water samples.
For the multihabitat sample, a dose from each single-
habitat sample was combined into one container.
16
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ANALYTICAL METHODS: ALGAE
Dr. Jan Stevenson of Michigan State University is
using three disturbance indicators: the land use in-
dicator developed by Maine DEP, trophic status
indicators (total nitrogen, total phosphorus, and
chlorophyll a), and hydrologic and sewage chemi-
cals (Ca, Na, Cl). Dr. Stevenson is comparing a
suite of algal indicators to determine which types of
indicators respond to the three disturbance indica-
tors. The algae indicators include biological integ-
rity measures such as genus-species richness,
Shannon diversity, and number of taxa in genera.
Dr. Stevenson is also using European algal
autoecology information to determine environmen-
tal characteristics for the taxa. This information will
give an autoecological index that shows a relation-
ship to variables such as moisture, organic N, low
oxygen, pH, salt, and nutrients. Although algae was
collected from all sites sampled between 1998 and
2002, only samples from 1998 and 1999 have been
processed to date because of funding limitations.
LESSONS LEARNED
Incorporating a wetland monitoring component
into Maine's exi sting biomonitoring program has
been an efficient way to pool limited resources
and build on Maine's successful river and stream
biomonitoring experience.
To implement a comprehensive biomonitoring
program for wetlands, Maine DEP needs to
build the capacity to assess multiple biological
assemblages, including algae and vascular
plants. This capability will improve the State's
ability to evaluate wetland impacts from stres-
sors such as nutrient enrichment and hydrologic
changes, and will allow for the assessment of
less frequently inundated wetland types where
aquatic invertebrates are not naturally abundant.
Current funding and staff levels prohibit expan-
sion of the program at this time.
17
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MARYLAND: DEVELOPING AN IBI ASSESSMENT FOR
RESTORED WETLANDS IN THE MID-ATLANTIC STATES
Contact
Peter Lowe
Organization USGSBRD
Patuxent Wildlife Research Center
11510 American Holly Drive
Laurel, MD 20708-4017
Phone (301)497-5705
E-mail peter_lowe@usgs.gov
Website http://www.pwrc.usgs.gov/wli/
PURPOSE
Develop sampling methods for different assem-
blages
Develop a yardstick of biological metrics to
assess the progress and condition of recon-
structed wetlands in Maryland, Delaware, and
Virginia
Compare the suitability of different assemblages
(plants, macroinvertebrates, amphibians) for
assessing wetland condition
Evaluate the sources and magnitude of variance
in data collected for biological metrics
Evaluate seasonal and annual biological fluc-
tuations for the wetland sites
WETLAND TYPE
DelmarvaBays (depressional wetlands with
emergent, scrub/shrub, or forested vegetation)
ASSEMBLAGES
Amphibians
Macroinvertebrates
Vascular plants
PROJECT HISTORY
The project is a joint effort among the USGS,
Patuxent Wildlife Research Center, theUSDANatu-
ral Resources Conservation Service Wetland Sci-
ence Institute, and EPA. Project development
started in 1995 in response to a lack of information
about the success of wetland mitigation projects,
especially those associated with wetland restora-
tion on farmlands under set-aside programs. From
1996 to 1998, fieldwork focused on a set of re-
stored and existing wetlands. In 1999, a second
set of wetlands was studied to evaluate the robust-
ness of the metrics developed from the first set. All
fieldwork has now been completed and metric
development is under way.
18
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STUDY DESIGN
Because one of the objectives was to assess the
sources and magnitude of variance in the different
measurements, we intentionally oversampled our
sites compared with what would be done under typi-
cal regional or statewide assessments. For 3 years,
work was done on a single set of 30 wetlands, in-
cluding 22 restored and 8 natural wetlands or
Delmarva bays. All wetlands in the database were
depressional, semipermanent, or seasonal.
Macroinvertebrates were sampled one to four times
each year, depending on hydrology; macrophytes
during spring and late summer; and amphibians al-
most continuously during the breeding season.
SAMPLING METHODS AND ANALYSIS:
MACROPHYTES
Data on plant species composition, abundance,
and dominance were collected through the use of
line transects. Pairs of 50 m transects were placed
on opposite sides of the wetland at four different
hydrological levels as determined at "full pool" lev-
els. The hydrological levels were buffer, less than
25 cm, 25 to 45 cm, and greater than 45 cm. In
wetlands larger than 5 acres, the number of transect
pairs was doubled. Each transect was divided into
five runs of 5 m each, alternating with 5 m of
unsampled transect. Along each run, sampling oc-
curred at specific points (as in point intercept meth-
ods) at 1-m intervals. Thus, each run of 5 m con-
tained five sample points. Each transect consisted
of five runs of five sampling points each. Each hy-
drological depth was sampled by 2 transects, bring-
ing the total number of sample points per hydro-
logical zone to 50. There were:
(4 hydrological zones) x (2 transects each zone)
x (5 runs per transect) x (5 points per run)
= (200 points per wetland)
This was done in spring and fall for each wetland.
At each point, the species name and number of in-
dividuals were recorded.
In addition to point data, incidental species along
the line were recorded to include the species not
intercepted at any points along the line. These spe-
cies were included in species richness and attributes
derived from presence or absence data, but not in
dominance calculations.
SAMPLING METHODS AND ANALYSIS:
MACROINVERTEBRATES
Aquatic invertebrates were sampled at approxi-
mately 6-week intervals beginning in late May and
continuing until October and were conducted in
association with the sampling of water quality,
aquatic plants, and hydrological and wetland dimen-
sions. Invertebrate samples were collected along
transects following compass coordinates originat-
ing from markers placed in the deepest part of ev-
ery wetland before the sampling seasons began.
Transect coordinates were randomly selected for
each wetland and sampling time. The method of
compass points was adapted to each wetland's
morphology.
Samples were collected from three depth ranges
along the transects to determine invertebrate rela-
tive abundance, diversity, and relative biomass in
each wetland. As long as water depths were
adequate, samples were collected from along the
transects at the following water depths: less than
15.0 cm, 15.1 to 45.0 cm, and greater than
45.1 cm.
Samples were collected using a modified Gerking
box sampler, which is a sheet aluminum box with a
sliding screen door (1-mm mesh) at the bottom.
The sampler has the advantage of allowing simulta-
neous collection of benthic, pelagic, neutonic, and
plant-associated invertebrates. The sampler was
lowered to the floor of the sampling area with the
screen door open. The vegetation in the sampler
was then cut at the mud-water interface and put
into prelabeled plastic bags. Then the screen door
was slowly closed as the sediments just in front of
19
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the advancing screen were stirred into the water
column. After the screen was closed, soil materials
were sieved through the screen by shaking the box.
All invertebrates caught on the screen were then
placed in prelabeled plastic bags. The bags of veg-
etation and invertebrates were placed on wet ice as
soon as possible and stored in a refrigerator until
the samples could be picked. Each individual was
keyed to the lowest taxonomic level feasible, typi-
cally species or genus. Data on species composi-
tion and abundance were used to generate metrics
in a manner similar to that for plants. An approved
quality assurance/quality control process was fol-
lowed, which included independent validation of
20% of our samples.
SAMPLING METHODS AND ANALYSIS:
AMPHIBIANS
Each site was sampled for amphibian larvae once
every 4 weeks. Sampled areas consisted of the
perimeter of the open-water portion of the wetland
and lightly vegetated areas that allowed a seine to
pass. We used a 6x8-m nylon mesh (1/16") seine
to sample each wetland by wading out 3 to 5 m
away from the shoreline and then moving in towards
the shoreline in one continuous sweep. Sampling
was time-constrained to 2 hours. If new species
were caught during the last two sweeps, the sam-
pling period was extended until no additional new
species were found. Amphibian larvae were iden-
tified using published keys and by temporarily hous-
ing tadpoles until they metamorphosed and could
be identified.
Drift fences were also used to supplement seining
data and obtain information on adults and
metamorphs. We initially considered surrounding
each wetland with a drift fence but realized the im-
practicality of that idea. Therefore, each wetland
was provided with 50-cm-tall and 15-m-long drift
fences. If possible, the fences were placed along
drains, travel corridors, and other likely points of
amphibian use in order to maximize capture of indi-
viduals entering and exiting the wetlands. Five-gal-
lon plastic buckets were buried in pairs at the ends
and midpoint on the inside and outside of the fence
to capture adults entering the ponds and juveniles
leaving the ponds. Wet sponges, rocks, and veg-
etation were placed in the buckets to prevent des-
iccation and provide some cover and refugia to cap-
tured individuals. All amphibians were identified,
sexed, and returned to the inside of the fence at the
wetland from which they were captured. Juveniles
and metamorphs departing the wetlands were iden-
tified, counted, examined for malformations, and
released on the outside of the drift fence.
GENERAL ANALYTICAL METHODS
A fundamental component of this study is to de-
vise a gradient of physical factors (e.g., land use in
drainage area, management techniques, landscape
features, method of restoration) that affect wetland
health. From there, data on the frequency of occur-
rence and relative abundance of species, guilds, or
trophic classes will be used to develop attributes
for an index of biological integrity (IBI) for each of
the assemblages. An attribute will be considered a
valid metric if it relates either positively or nega-
tively to the physical gradient. We will then com-
pare the IBIs developed and determine if they are
consistent in ranking wetlands. Once acceptable
IBIs have been developed on the initial wetland
base, we will apply them to the second set of wet-
lands to validate the model. In addition, the vari-
ance within our sampling methodology will be as-
sessed.
LESSONS LEARNED
Overall
Many of the wetlands in our bases were only a
few years old when we started and may not
have had sufficient time for ecological and an-
thropogenic factors to separate them along a
physical gradient.
2O
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As a result, development of a reliable and eco-
logically meaningful gradient has been one of
the most difficult parts of this proj ect.
It would be very instructive to revisit these wet-
lands after 10 or 15 years and see how they
have changed.
Macrophytes
There can be considerable differences between
mid- and late summer in the ability to easily
record and identify plants. This is especially true
for graminoids, which are primarily identified
by fruiting body characteristics. In addition,
many legumes, composites, and warm-season
grasses are present in late summer but not ap-
parent in spring.
The inclusion of incidental species added ap-
preciably to the number of species identified in
a particular wetland. We are evaluating whether
this inclusion has an effect on the resulting
metrics.
Deep-water areas (greater than 45 cm) have
much lower species richness than shallower
zones and do not need to be sampled at the
same level of intensity at the same site.
Permanent transects are preferred if data col-
lection can continue over several years. This
will allow for the annual and seasonal changes
that occur over time through shifts in hydrol-
ogy.
Macroinvertebrates
Picking, sorting, and identifying aquatic insects
was one of the most laborious aspects of the
study. Investigators wishing to use invertebrates
in their bioassessments should allocate sufficient
resources to accomplish the task.
Invertebrate species presence, and especially
abundance, are seasonally quite variable. June
to early July before the drying of mid- to late
summer begins seems to be the best months for
finding the greatest diversity and abundance of
macroinvertebrates.
AMPHIBIANS
The reduced species richness of amphibians,
compared to macrophytes and macro-
invertebrates, may limit the number and types
of metrics that can be developed from this
assemblage.
Adequate sampling for amphibians requires
more trips and techniques than other assem-
blages. This is due to their mobility, multiple life
history strategies, and variable breeding peri-
ods among species. Sampling for one life stage
only is probably not as effective as sampling for
adults and tadpoles in determining amphibian
usage of a wetland.
21
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MASSACHUSETTS: USE OF MULTIMETRIC INDICES TO
EXAMINE ECOLOGICAL INTEGRITY OF SALT MARSH
WETLANDS IN CAPE COD
Contact
Bruce K.Carlisle
Organization Massachusetts Coastal Zone Management
251 Causeway Street
Boston, MA 02114
Phone (617) 626-1200
E-mail bruce.carlisle@state.ma.us
Website http ://www. state.ma.us/czm/wastart.htm
PURPOSE
r I The primary focus of the Cape Cod Salt
J. Marsh Assessment Proj ect is to advance and
improve the salt marsh assessment approach de-
veloped by the Massachusetts Coastal Zone Man-
agement (MA CZM) team through the application
and review of the existing protocol in two different
assessment investigations. The first investigation,
conducted in the 1999 field season (May to Octo-
ber index period), examined salt marsh indicators
from six sites on the Cape Cod Bay coast; these
sites had varying types and intensities of human land
use or disturbance. The second investigation is a
long-term comparison of indicators from three pairs
of salt marshes, each pair having a marsh area with
restricted tidal hydrology and a corresponding area
with normal tidal hydrology. The intent of this work
is to document differences in indicators before and
after tidal restoration actions. Through the imple-
mentation of these two investigations additional
obj ectives will be realized. The collection and com-
pilation of data on the condition of relatively undis-
turbed sites is of critical importance to the evalua-
tion and determination of impaired sites. This
proj ect will serve to expand the salt marsh refer-
ence site database. Another important aspect of
this proj ect will be to further examine the suite of
existing metrics and attributes used for biological
comparison and to explore new metrics, based on
the proj ect data and literature/information base. The
long-term tide restriction study will provide insight
as to the utility of this assessment approach as a
measure of determining salt marsh restoration
progress and trajectory.
WETLAND TYPE
Salt marshes
ASSEMBLAGES
Birds
Fish
Macroinvertebrates
Vascular Plants
STATUS
The Cape Cod Salt Marsh Assessment Proj ect is
approximately two-thirds complete, with the land-
use investigation finished and the longer-term tide
restriction study entering its third year (or index sea-
22
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son). The report for the 1999 land-use investiga-
tion is being finalized and should be available in early
fall 2002 from this site or MA CZM's site (http://
www.state.ma.us/czm/wastart.htm). EPA grant
funds will support two additional years of field data
collection, with project completion anticipated in
March 2004. Field investigations from 2000 through
2004 are focused on the assessment of the effects
of tidal hydrology alterations and the restoration of
normal hydrology on salt marsh ecological integrity.
The study involved three salt marsh sites with tidal
hydrological restrictions. Of the project sites, one
(EOBP) received restoration action in spring 2002,
one (EMC) is predicted for fall 2002, and one
(MSLP) is still uncertain for restoration action, with
unforeseen complexities. In addition, the project
team expects to sample one additional reference
site each year to expand the regional reference
dataset.
PROJECT DESCRIPTION
Massachusetts Coastal Zone Management
launched its Wetland Assessment Program with its
first effort in wetland bioassessment in 1996-1998
at the Waquoit Bay National Estuarine Research
Reserve. A subsequent proj ect was implemented
on Massachusetts' North Shore in 1997-1999.
The EPA Region I sponsored pilot, Cape Cod
Salt Marsh Assessment Proj ect, began in 1999 and
will continue through 2004. The investigative team
for this proj ect includes Bruce Carlisle, Tony Wilbur,
Jan Peter Smith, and Jay Baker from MA Coastal
Zone Management, and Anna Flicks, an indepen-
dent consultant specializing in aquatic
macroinvertebrates. Partners for thi s proj ect in-
clude the MassBays Program, UMass Extension,
the MA Department of Environmental Management
(DEM) and its Area of Critical Environmental Con-
cern Program, the Waquoit Bay National Estuarine
Research Reserve, the Cape Cod Commission, and
the towns of Barnstable, Eastham, Mashpee, Or-
leans, and Sandwich.
The Massachusetts CZM proj ect team will apply
standardized sampling and surveying protocol to salt
marsh study sites to gather biological, chemical, and
physical data for the two investigations referred to
earlier.
The proj ect team will analyze and express the bio-
logical data through a series of existing metrics or
attributes, or develop new metrics or attributes as
necessary, based on the project data and literature
base. Chemical and physical data will be utilized as
supporting information sources.
The team will make recommendations for revi-
sions, additions, or deletions to its current wetlands
assessment approach.
STUDY DESIGN
In the 1999 field season, six salt marsh sites with
varying types and degrees of intensity of surround-
ing land use were selected. Two sites with minimal
human land use (conservation land and no tidal hy-
drological alteration) were chosen as reference
sites, representing the best attainable conditions in
the immediate region.
The four other salt marsh study sites have varied
land uses including residential, commercial, and
transportation. The impacts associated with these
land uses include direct stormwater outfalls, large
impervious areas, septic systems, lawn fertilizer/
chemicals, pet waste, automobile emissions/
byproducts, and direct habitat alterations.
For the 2000 field season, three salt marshes with
tidal hydrological restrictions will be studied. Mea-
surements will be made both at the salt marsh af-
fected by the tidal restriction (the restricted study
site) and at the salt marsh below the restrictive fea-
ture (the reference site). The reference sites or
marshes below the tidal restriction receive normal
tidal influence and inundation. In addition, the 2000
field season will also include two additional long-
23
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term salt marsh reference sites. The continued de-
velopment of a robust reference site database is
critical to the continuing evolution and application
of wetland bioassessment.
SAMPLING METHODS:
MACROINVERTEBRATES
At each site, a habitat characterization form will
be completed that summarizes the ambient salt
marsh habitat conditions at the study site. The in-
formation collected includes water quality param-
eters (temperature, pH, dissolved oxygen, salinity,
and color) and habitat descriptors on hydrology,
vegetation, substrate, available food sources for in-
vertebrates, and degree of human impact.
The sampling protocol will utilize tree devices for
application in different habitats:
Intertidal zone
Plot sampling using wooden frame (18" x 18")
Subtidal (permanently flooded) zone
D-net, plot sampling, and auger
At each sampling site, three sampling stations will
be selected over a defined linear distance of the
creek channel, stations will be located at 1/3 inter-
vals. At each of the three stations a representative
composite sample of macroinvertebrates will be col-
lected as follows:
Intertidal bank zone at low tide: 1 plot sample
Subtidal estuarine zone at low tide: 1 plot
sample, 1 D-net sample, and 1 auger sample
Each sample will be placed in a zip lock bag la-
beled with site number, site name, date of sampling,
sample number, sampling method, and name of sam-
pler. The site field sheets also record the relevant
sample numbers. Samples will be preserved in 70%
isopropenol alcohol and placed in a cooler ready
to be transported to the laboratory for sorting and
identification.
Invertebrate samples are taken once in May and
again in August.
SAMPLING METHODS: VEGETATION
At each site, the salt marsh wetland vegetation
will be surveyed according to this protocol. Six
transects will be established based on a stratified
random sampling approach. A defined linear dis-
tance of the salt marsh creek channel is established.
The evaluation area will be segmented into three
subunits along equal sections of the creek channel.
The first third of this length is subunit #1, the sec-
ond third is subunit #2, and the final third is subunit
#3. In each of the subunits, two randomly selected
transects will be laid. The transect locations will be
determined by a computer random numbers
algorithm.
The transects will run roughly perpendicular from
the channel to the upland edge, and each transect
will be laid according to a consistent compass bear-
ing. Along each transect, 1-m2 quadrats will be
located every 60 feet, starting at the creek edge
and progressing along the entire length of the transect
until the upland edge. The last quadrat will be lo-
cated in the salt marsh fringe community, well within
the wetland and not on the upland.
Using a standard data sheet, in each quadrat along
each transect, every plant occurring within that quad-
rat will be identified by genus and species. For
each unique species within the quadrat, the abun-
dance of that species will be determined using stan-
dardized coverage charts. Investigators will also
define the community type that the quadrat falls in:
low marsh, high marsh, or fringe. To be as accu-
rate as possible, coverage estimates include duff,
leaves, bare ground, and open water, collectively
designated as "other." Coverage estimates will be
adjusted during the analysis stage to account for
the coverage of this "other" category.
24
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Vegetation surveys will be conducted once at each
site during the peak growing period from mid-July
to mid-September.
SAMPLING METHODS: BIRDS
Point counts will be utilized as the primary sam-
pling method, using visual and auditory cues. At
least two expert observers, including the principal
investigator, will sit quietly from a vantage point
where all of the evaluation area can be viewed.
Using a standard data sheet, all species and indi-
viduals will be counted and recorded by the ob-
servers, as they are heard or seen demonstrating
any activity in the evaluation area or in a 100-foot
buffer area surrounding the evaluation area. Counts
will be conducted for a period of 20 minutes, sepa-
rated into four 5-minute sample intervals. All indi-
viduals will be counted, with a concerted effort not
to duplicate individuals. An additional 10 minutes
will be allotted to allow observers to walk slowly
along the perimeter of the wetland in order to de-
tect any species not tallied in the 20-minute count.
Several sites may be visited on the same day, with
census beginning at approximately 6 am and ceas-
ing at approximately 8:30 am in order to capture
peak activity. Sites will be sampled in late August
to capture migrating shorebird usage, as saltmarsh
habitats are known to have comparatively fewer
breeding species.
SAMPLING METHODS: FISH
The sampling strategy will be to capture the chan-
nel habitat (sub- and intertidal). As detailed above,
the evaluation area will be divided into three sub-
units along a defined length of creek channel. Sta-
tions will be established for each habitat as follows.
In the creek channel, fixed stations will be deter-
mined by a computer random numbers algorithm
producing a random integer between 0 and 100.
The random integer will be the distance of the start-
ing point for the seine haul in feet from the start
point (0') of each subunit.
Seines will be utilized to sample the creek chan-
nel. At three stations, as defined above, the seine
will be dragged through the water column along the
creek bank and substrate for a length of 5 meters.
Seines will be carefully withdrawn from the creek
and the collected fish will be carefully extracted from
the seine into a processing bucket. The fish will
then be removed by dipnets and individuals will be
identified and measured. Abundance and total bio-
mass by species will be enumerated. All data are
recorded on a standardized field sheet.
One sample run will be conducted each month
from April to October.
ANALYTICAL METHODS
Biological data collected at wetland study sites
are compared to data collected at the wetland ref-
erence sites. Multimetric data analysis techniques
are employed to examine attributes and variables
of biological data, and these metrics are combined
into a quantitative final index. Ametric is a param-
eter or variable that represents some feature, sta-
tus, or attribute of biological assemblage, chemical
state, or physical condition. In a multimetric ap-
proach, several different metrics are chosen in or-
der to effectively capture and integrate information
from individual, population, guild, community, and
ecosystem levels and processes. Metrics are se-
lected on the basis of literature reviews, historical
data, and professional knowledge. The quantita-
tive output from each metric is then combined to
produce an index. An index is the aggregate of
weighted metric scores that serves to summarize
the biological condition.
LESSONS LEARNED
Through the three pilot proj ects, the MA CZM
project team has been able to learn from each ap-
plication and as a result has made several small,
incremental revisions to many components of the
protocols, including adjustments to sampling meth-
25
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ods and shifts in the attributes and metrics exam-
ined. Each application has also generated results
that indicate decreasing biological integrity with in-
creasing land-use stressors.
Results from 6 years of wetland assessment
experience have led the MA CZM team to con-
clude that this assessment approach has significant
potential for a number of management applications,
including:
Inventory efforts such as identifying and pro-
tecting unique and important habitat
Identification of degraded salt marsh sites and
presence of invasive (nonnative) species in or-
der to evaluate restoration potential and/or re-
port on wetland status (i.e., 305b reports)
Monitoring and evaluating the effectiveness of
restoration actions
Long-term tracking of salt marsh condition and
documenting the effects of disturbances (i.e.,
stormwater pollution, eutrophication,
hydromodifi cation)
Many of these applications have appeal to water-
shed-based organizations and agencies as well as
volunteer groups. The techniques of the MA CZM
salt marsh assessment approach have been success-
fully taught through a comprehensive volunteer train-
ing program on Massachusetts' North Shore. At
the time of writing, the Wetland Health Assessment
Toolbox program is entering its fourth year, guiding
and supporting volunteer groups as they monitor
salt marsh restoration sites in their region (http://
www.salemsound.org/wetlands.htm).
Results from this Cape Cod Salt Marsh Assess-
ment Proj ect are still being analyzed and written
up, but some initial details are available: The 1999
land-use investigation has confirmed the trend seen
in the previous two studies where increasing levels
of land use around a given salt marsh site (wetland)
correspond with decreased ecological condition, in
this case indicated by the average of the Plant Com-
munity Index and the Invertebrate Community
Index.
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FIGURE 7: RELATIONSHIPS BETWEEN WETLAND ECOLOGICAL CONDITION AND
LAND USE. THE LAND USE INDEX (LUI) SCORE is A MEASURE OF HUMAN
DISTURBANCE. WITH INCREASING LAND USE TYPES AND INTENSITIES, THE LUI
SCORE DECLINES. SIMILARLY, THE WETLAND ECOLOGICAL CONDITION
(WEC) IS A MEASURE OF BIOLOGICAL INTEGRITY. A LOWER WEC SCORE
INDICATES INCREASING BIOLOGICAL IMPAIRMENT.
27
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Abn. Habitat Affin.
EBAbn. Opportunistic
DAbn. PSL
DAbn. Salinity
DAbn. Nutrient Affin.
Abn. Invasive
DTaxa Richness
Community Similarity
FIGURE 8: Two SETS OF MULTIMETRIC GOALS FOR EVALUATING RESTORATION.
THE RESTORED SITE IS ON THE BOTTOM, ITS PAIRED REFERENCE (DIRECTLY
BELOW/SEAWARD TIDE RESTRICTION FEATURE) IS THE MIDDLE GRAPH, AND A
REGIONAL REFERENCE MARSH IS DISPLAYED AT TOP.
28
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PLATE 5: CAPE COD, MA, SALT MARSH PLANT SURVEY:
1 M2 ALONG TRANSECT. PHOTO! B.K. CARLISLE
PLATE 6: CAPE COD, MA, SALT MARSH NEKTON: WATCH YOUR FINGERS!
(BLUE CRAB, CALUNECTESSAPIDUS). PHOTO: B.K. CARLISLE
29
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MASSACHUSETTS: INVOLVING VOLUNTEERS IN
EXAMINING THE ECOLOGICAL INTEGRITY OF COASTAL
WETLANDS IN CAPE COD, MASSACHUSETTS
Contact
BrittaMagnuson
Organization Salem Sound 2000
201-203 Washington St., Suite 9
Salem, MA 01970
Phone (508) 741-7900
E-mail Brita.Magnuson@salemsound.org
Contact
Jan Smith
Organization Mass Bays National Estuary
Program
251 Causeway Street, Suite 900
Boston, MA 02114
Phone (617)626-1231
E-mail jan.smith@state.ma.us
Website http://www.state.ma.us/czm/
wastart.htm
PURPOSE
Training volunteers to do biological assessments
Comparing results of volunteers to trained sci-
entists
WETLAND TYPE
Salt marshes
ASSEMBLAGES
Birds
Fish
Macroinvertebrates
Vascular plants
STATUS
Completed 3 years of sampling. Analyzing data.
PROJECT DESCRIPTION
"Salem Sound 2000" and "8 Towns and the Bay,"
two regional subcommittees of the Massachusetts
Bays National Estuary Program, participated in a
salt marsh monitoring pilot proj ect involving citizen
volunteers in conjunction with UMass Cooperative
Extension Service and Massachusetts Coastal Zone
Management (MCZM).
During the summers of 1999 and 2000, more than
40 volunteers participated in training workshops and
field data collection for a variety of parameters:
water chemistry, land use index (a habitat assess-
ment), aquatic macroinvertebrates, birds, tidal in-
fluence, and vegetation. During 1999, professional
scientists did independent assessments, and the
volunteers conducted assessments (using the same
sampling protocols as the professionals) with the
guidance of trained staff members. Data compari-
sons, as well as feedback from volunteer partici-
pants, were used to modify and improve training
protocols for the 2000 field season. Field data were
collected in 2000 at the same sites, which included
3O
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salt marsh sites that were affected by tidal restric-
tions and sites impacted by stormwater discharges.
The project is expected to be a model in other ar-
eas of New England. A volunteer training manual is
currently being printed and should be available at
the time of this printing. Funding from a private
foundation has enabled the program to continue for
two additional years, 2001 and 2002, and it is
planned to add fish as an additional parameter for
the volunteers to measure. The sampling proto-
cols are summarized in the MCZM project sum-
mary.
LESSONS LEARNED
Volunteers may need ongoing field instruction in
order to ensure quality data collection. The initial
comparison between data collected by volunteers
and data collected by professionals indicated gaps,
which improved field assistance was able to reduce.
Also, asking the volunteers to review and comment
on training workshops was helpful in improving pro-
gram design. Although there is high turnover in vol-
unteers from year to year, we are finding, after 4
years into the program, that some volunteers do
come back after a year's break, and that the skills
of the returnees definitely improve after the first year.
Offering teacher training credits improves the inter-
est of educators in participating in the program, and
many get interesting ideas for future use in their class-
rooms.
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MICHIGAN: BIOASSESSMENT PROCEDURES AND
BASELINE REFERENCE DATA FOR GREAT LAKES
COASTAL MARSHES AND INLAND FORESTED
WETLANDS IN MICHIGAN
Contact
Thomas M. Burton
Contact
Donald G Uzarski
Organization Michigan State University
Department of Zoology
203 Natural Science
Last Lansing, MI 48824-1115
Phone (517)353-4475
E-mail burtont@msu.edu
PURPOSE
Subproject 1: Coastal wetlands
Collect baseline data on water quality and ad-
jacent land use, as well as plant, invertebrate,
and vertebrate communities from Great Lakes
coastal wetland sites experiencing a continuum
of disturbance
Continue development of invertebrate and fi sh-
based indices of biological integrity (ffil's) by
plant zone, for Great Lakes coastal wetlands
Continue testing and validation of our IBIs
Subproject 2: Inland, forested depressional
wetlands
Collect baseline data on plant, invertebrate, and
vertebrate communities with accompanying
chemical/physical parameters from reference
and impacted forested, depressional wetlands
of southern Michigan
Develop an IB I for forested depressional wet-
lands of southern Michigan based on inverte-
brates, plants, fish (if present), and birds
Organization Grand Valley State University
Annis Water Resources Institute
Lake Michigan Center
740 West Shoreline Dr.
Muskegon, MI 49441
Phone (616) 895-3989
E-mail uzarskid@gvsu.edu
WETLAND TYPE
Great Lakes coastal
Inland, forested depressional
ASSEMBLAGES
Birds
Fish
Macroinvertebrates
Vascular plants
STATUS
Completed 3 years of sampling. Analyzing data.
PROJECT DESCRIPTION
Subproject 1: Coastal wetlands
We developed a preliminary IBI for Lake Huron
based on invertebrate data collected from coastal
wetlands with funds provided by the Michigan De-
32
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partment of Environmental Quality, the Nature Con-
servancy, U.S. EPA, and the U.S. Geological Sur-
vey and published it in the j ournal Wetlands (Bur-
ton etal. 1999). Since publication, we have con-
tinued testing the IBI with data from additional
Lakes Huron and Michigan fringing wetland sites.
We also have continued monitoring a set of initial
sites used in IBI development as lake levels de-
clined in order to determine whether and how well
the IBI works as lake levels decline substantially
below levels that occurred during IBI development.
We have collected extensive data on invertebrates,
plant, fish, and bird communities on Lake Huron
wetlands since the early 1990's. Much of our work
has been in collaboration with the Great Lakes Sci-
ence Laboratory (BRD-USGS), Michigan Natural
Features Inventory (MNFI), and the Ohio Biologi-
cal Survey (OB S).
Our preliminary IBI for Lake Huron fringing wet-
lands was developed using macroinvertebrate data
collected in 1997 from six wetlands. It was tested
using data collected from 11 Lake Huron wetlands
(6 original and 5 additional) in 1998 at lake levels
substantially lower than in 1997. We continued to
test the IBI using data collected from 12 sites (7
original and 5 additional) at even lower lake levels
in 1999 and from 5 additional sites at extremely
low water levels in 2000. Even though some plant
community zones used in IBI development were
not flooded and could not be sampled in 1999 and
2000, the IBI functioned extremely well. In 2001,
we sampled similar fringing wetlands of northern
Lake Michigan along with northern Lake Huron sites.
The IBI appears to work for fringing wetlands of
northern Lake Michigan as well.
While testing and validating the Lake Huron IBI
in 1999,2000, and 2001, we developed modifica-
tions to simplify and improve it. A detailed expla-
nation for the modifications was presented at "Wet-
lands 2000" in Quebec City in August 2000, and
additional modifications to remove the "preliminary"
status were presented in Lake Placid, New York,
in June 2002. The modifications are as follows:
1. The Typha zone should be removed from the
mi.
2. The four diversity and richness metrics should
be calculated by plant zone instead of combin-
ing data from all plant zones before calculations
are made.
3. Two new metrics should be added to the Inner
Scirpus Zone:
- Relative abundance Isopoda (%)
Decreases with disturbance
- Relative abundance Amphipoda (%)
Increases with disturbance
4. Use !/2 person-hour count to determine num-
ber of individuals counted per replicate. Count
either 50, 100, or 150.
A manuscript noting all of the above improvements
is in preparation and will be submitted to the jour-
nal Aquatic Ecosystem Health and Management
by September 2002 as part of a special issue on
coastal wetlands.
We are optimistic that our Lake Huron IBI will
work for northern Lake Michigan fringing wetland
sites. These sites include most wetlands along the
southern shore of the Upper Peninsula of Michigan
from St. Ignace to the Wisconsin border. Most of
these wetlands appear to be comparable in plant
community composition and structure to the Lake
Huron wetlands. Preliminary testing took place in
2001 and we expect to be able to recommend the
ffil's use for these types of wetlands.
In 2000, we began development of new fish- and
macroinvertebrate-based ffils for the drowned river
mouth wetlands of Lake Michigan. We collected
data from eight sites representing a gradient of an-
thropogenic disturbance. Ten sites were sampled
in 2001 and several new sites will be added in 2002.
We expanded our work on drowned river mouth
wetlands in 2001 to Lake Superior wetlands.
33
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Subproject 2: Inland, forested depressional
wetlands
We started our forested depressional wetland
proj ect in 1999. Invertebrates were collected three
times per year from eight forested depressional
wetlands during 1999 and 2000 using dipnets, ac-
tivity traps, and black lights. Additional samples
were collected from a subset of these and some
additional wetlands in 2001. Up to 40 additional
sites will be added in 2002, with emphasis on ob-
taining a greater array of impacted sites and ex-
tending the sites to isolated depressional wetlands
in the coastal zone of the Great Lakes. Inverte-
brates are identified to the lowest taxonomic unit
possible, and preliminary ffil development has be-
gun. Accompanying chemical/physical samples
taken from surface water and mini-piezometers have
also been recorded and analyzed for potential
metrics. Comparisons of the plant and invertebrate
communities have been made using a subset of the
sites.
A list of potential metrics, a summary of our very
preliminary analyses, and conclusions that we drew
from the 2000 data set are as follows:
Significantly higher at reference sites (Mann-
Whitney tests):
Isopoda p< 0.001
p = 0.004
Amphipoda
Diptera
Culicidae
Ephemeroptera
Trichoptera
p = 0.004
p = 0.015
p = 0.078
Significantly higher at impacted sites (Mann-
Whitney tests):
Gastropoda
Lymnaeidae p = 0.004
Planorbidae p< 0.001
Diptera
Chironomidae
Chaoboridae
Odonata
Libellulidae
Hemiptera
Lleididae
Coleoptera
Halipidae
Annelida
Hirudinea
p = 0.074
Collected from only one
site
p = 0.039
p< 0.001
p< 0.001
Collected from only one
site
SUMMARY
Of the measured chemical/physical variables, only
depth separated impacted and reference sites.
Canopy cover was not measured in 1999 and 2000
but may have also separated impacted and refer-
ence sites. Estimates of canopy cover are currently
being obtained for all sites. Eleven (+2 ?) taxa
showed potential as metrics, but not during early
inundation (April). During full inundation (May),
the correspondence analysis grouped the wetlands
using invertebrate community composition into dis-
tinct categories.
Surface runoff-influenced impacted sites
Groundwater-influenced reference sites
Precipitation-influenced reference sites
CONCLUSIONS
Increased runoff may increase water depth and
subsequently kill trees, opening the canopy.
- Some taxa such as Libellulidae may be
responding to these changes in ambient
conditions.
34
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Most traditional chemical/physical variables will
not be useful as indicators of anthropogenic dis-
turbance in these wetlands. These variables in-
clude turbidity, DO, pH, SRP, NH4, conduc-
tivity, alkalinity, temperature.
An invertebrate-based IBI for (hardwood) for-
ested wetlands of Southern Michigan appears
to be feasible.
STUDY DESIGN
Subprojects 1 and 2
Sites that experience a wide range of anthropo-
genic disturbance, or stressors, are chosen from each
hydrogeomorphic class or subclass of wetland. The
extent of disturbance is determined using surround-
ing land use/land cover and limnological data. Ini-
tially, correspondence analyses of invertebrate com-
munity composition were used to determine if ref-
erence sites separated from impacted sites. When
they did, individual taxa containing the most inertia
responsible for the separation were deemed po-
tential metrics. Mann-Whitney U tests were then
used to determine if densities of these taxa at refer-
ence sites were significantly different from densities
at impacted sites.
We use medians in place of means in the IBI be-
cause medians are more resistant to the overwhelm-
ing effects of outliers. Our goal is to typify the wet-
land. If an area is sampled that is depleted or con-
centrated in the constituents of a metric, the area
may be isolated from anthropogenic disturbance,
receiving a dose of disturbance not typical of the
entire wetland or vegetation zone, or may contain
some "natural" chemical/physical component that
is unique. Regardless of the cause, the area is not
representative of the entire wetland. The influence
of these outliers can be dampened by using the
median in place of mean as a measure of central
tendency.
After potential metrics were developed, principal
components analysis (PCA) was used to establish
principal components (PCs) based on chemical/
physical parameters as well as surrounding (1-km
buffer) land use/cover data. Pearson correlations
were done between individual metrics and PCs to
establish stressor-ecological response relationships.
PCs were then decomposed to explore relative
contributions of individual stressors.
SAMPLING METHODS
Subprojectl: Coastal Wetlands
Macroinvertebrate samples. Macroinvertebrate
samples were collected with standard D-frame,
0.5-mm mesh dipnets. All maj or plant community
zones were sampled at each site, including a deep
emergent and a shallow, wet meadow zone. If cer-
tain depths contained more than one dominant plant
community along the shoreline, each plant commu-
nity type was sampled.
Dipnet sampling entailed sweeps through the sur-
face and middle of the water column and above the
sediment surface to ensure that an array of micro-
habitats were included in each sample. Dipnets were
emptied into white enamel pans, and 150 inverte-
brates were collected by removing all specimens
from small areas of the pan. Special consideration
was made to ensure that smaller organisms were
not missed, as there is a bias toward larger, more
mobile individuals with this technique. Plant detri-
tus was left in the pan and sorted through for a few
additional minutes to ensure that sessile species were
included in the sample. If 150 individuals were not
obtained after Va person-hour of field picking, we
collected to the next multiple of 50. The timed count
was used to semi quantify sampling effort so that it
could be used as a metric. Three replicate samples
were collected in each plant zone to obtain a mea-
sure of variance.
Dipnet samples were collected from late July
through August. Samples taken from ice-out through
mid-July generally contained less diversity and a
greater proportion of early instars of aquatic insects,
35
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making identification more difficult. The July-
August time period also corresponded to the time
when plant communities, characteristic of these
wetland systems, achieved maximum annual
biomass.
Fish sampling. Six fyke nets (0.5-in mesh) were
set in each wetland for 1 net-night. Nets were set
adj acent to specific plant zones with the leads bi-
secting the sampling area. Minnow traps were used
as a secondary method.
Subproject 2: Inland, forested depressional
wetlands
Invertebrate sampling. Macroinvertebrate
samples were collected with standard D-frame
dipnets containing a 0.5-mm mesh. Three replicate
samples were collected from three locations in each
wetland: (1) the deepest portion (usually the cen-
ter), (2) near the upland, and (3) between these
two areas. We attempted to incorporate habitat
heterogeneity by sampling as many plant zones as
possible at each location.
Dipnet sampling entailed sweeps through the wa-
ter column at the surface, middle of the water col-
umn, and above the sediment surface. Dipnets were
emptied into white enamel pans and 150 inverte-
brates were collected by focusing on small areas of
the pan and removing all specimens. If 150 indi-
viduals were not obtained after !/2 person-hour of
field picking, we collected to the next multiple of
50. The timed count was used to semiquantify sam-
pling effort so that it could potentially be used as a
metric. Invertebrate sampling was conducted dur-
ing early (April), full (May), and late inundation
(June) at each site.
Fish sampling. The temporary pools that domi-
nate most depressional forested wetlands are un-
likely to contain fish. Thus, fish sampling was only
conducted for permanent pools in depressional
wetlands when they occurred. Small fish traps were
placed in each of the permanent pools.
Birds surveys. Bird communities were surveyed
using 10-minute, 50-m radius point counts (Ralph
et al. 1995) by dual observers to sample the bird
communities at each count location. The 10-minute
counting period began when the observers reached
the perimeter of the 50-m radius so that any birds
flushed or silenced by the observer's approach
were detected (Riffell et al. 1996). The bird com-
munities of 30 forested wetlands were sampled in
2000 using a total of 6 count visits. A variety of
habitat measures from each site were obtained as
detailed under plant sampling below, and a manu-
script on results is currently being prepared. No
new sampling has occurred since 2000.
Plant sampling. The plant community and other
habitat variables were described for each 50-m ra-
dius bird count area along four habitat sampling ra-
dii radiating from the center of each point count sta-
tion following procedures detailed in Riffell et al.
(1996). The understory habitat was described us-
ing a Wien's pole (Rotenberry and Wiens 1985)
following procedures of Riffell et al. (2001), modi-
fied as appropriate to adapt them to forested
habitat.
More traditional plant sampling, of a subset of
forested wetlands, was done by Mike Kost and
Dennis Albert of the Michigan Natural Features In-
ventory during 1999 and 2000 using quadrat sam-
pling along transects running through each wetland.
They have submitted a report on their results to the
Michigan Department of Environmental Quality in-
cluding recommendations on potential metrics for
these wetlands.
36
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MINNESOTA: DEVELOPING WETLAND BIOCRITERIA
Contact
Mark Gernes
Contact
JudyHelgen
Organization Minnesota Pollution Control
Agency (MFC A)
Environmental Outcomes Division
520 Lafayette Road
St. Paul, MN 55155
Phone (651)297-3363
E-mail mark.gernes@pca.state.mn.us
Website http://www.pca.stat.mn.us
Organization Minnesota Pollution Control
Agency (MFC A)
Environmental Outcomes Division
520 Lafayette Road
St. Paul, MN 55155
Phone (651)296-7240
E-mail judy.helgen@pca.state.mn.us
Website http://www.pca.state.mn.us
PURPOSE
Develop the tools to assess wetland condition
by studying vegetation and invertebrates in wet-
lands across a range of human disturbance
Develop two separate Indexes of Biological
Integrity (IBI) for Minnesota depress!onal wet-
lands
WETLAND TYPE
Depressional wetlands, large and small; ripar-
ian wetlands
ASSEMBLAGES
Macroinvertebrates
Vascular plants
STATUS
Established indexes of biological integrity (IBIs) for
plants and macroinvertebrates. Report completed
on IBIs for 44 large depress!onal wetlands in May
2002. In 2002, evaluating data on statistical analy-
sis of sampling methods in 9 (invertebrates) and 12
(vegetation) large depressions. Beginning June
2002, testing IBIs in new ecoregion, comparing IBI
assessments with rapid assessment method and citi-
zen monitoring data. Training of many citizen com-
munity teams in biological assessment of local wet-
lands continued in 2002; a pictorial guide will be
produced in 2002.
PROJECT DESCRIPTIONS
Minnesota has been developing its wetland bio-
logical assessment program since 1992.
1992 - Small depressional reference wetland
study
The first proj ect was funded by the State legisla-
ture to develop biological reference conditions for
small depressional wetlands in central Minnesota.
Subsequent funding is primarily from EPA. The ini-
tial research studied the quantity and quality of
macroinvertebrates primarily in highest quality, least
disturbed reference sites and relatively few disturbed
depressional wetlands.
1995 - Expanded small depressional wetland
study
A second proj ect funded by EPA was undertaken
to develop multimetric IBIs for depressional wet-
lands in central Minnesota. During the 1995 sam-
pling season, MPCA collected data on macro-
invertebrates and vegetation for a larger set
of depressional wetlands than the 1992 study, rep-
37
-------
resenting a wide range of impairment. MPCA's
report on the 1995 ffil development was completed
in 1999 and will be available on MPCA's web site
(www.pca.state.mn.us).
1999 - Large depressional wetland study
MFC A sampled large depressional wetlands, in-
cluding duplicate sampling at six of the sites, to vali-
date metrics developed with smaller depressional
wetlands in 1995. MFC A also developed a scor-
ing system for human disturbance (HDS) that in-
corporates estimates of disturbances in the buffer
area and in the near-wetland landscape, plus ranges
of chemical pollution and alterations within the wet-
land. The HDS scores were used as the x-axis to
calibrate the metrics and the ffil scores. Both the
vegetation and the invertebrate ffil scores show sig-
nificant relations to the human disturbance scores
and to various water and sediment chemical fac-
tors. The invertebrate ffil was significantly related
to HDS, turbidity, phosphorus, chloride, and other
factors. The vegetation ffil was significantly re-
lated to HDS, phosphorus, and chloride in water;
and to copper; zinc; and nickel in sediments. Mi-
nor changes were made in metrics compared with
those used for the smaller depressions. The final
report for this proj ect, Indexes of Biological Integ-
rity (ffil) for Large Depressional Wetlands in Min-
nesota, by M.C. Gernes and J.C. Helgen, was com-
pleted in May 2002. In 1999, 27 wetlands in
riparian areas of small and medium-sized streams
in the St. Croix River basin were sampled for the
vegetation ffil. There data are currently under
analysis.
2001 - Statistical assessment of wetland
monitoring methods
MFC A sampled nine wetlands in three locations
to test the methods for the invertebrate and vegeta-
tion ffils with statistical procedures. These data
are currently under analysis.
2002 - Citizen training
In 2002, MFC A expanded efforts for training citi-
zens in biological assessment of wetlands, training
90 citizens in the invertebrate and vegetation meth-
ods (see Minnesota case study entitled "Dakota
County Wetlands Health Evaluation Proj ect"). A
final guide for biological monitoring of wetlands by
citizens will be produced by MFC A in 2002. This
guide includes pictorial keys to wetlands inverte-
brates and plants. MPCA also trained school teach-
ers in the assessment of ephemeral wetlands in the
spring of 2001 and 2002.
In the summer of 2002, MPCA is validating the
ffils for depressional wetlands in 40 wetlands in
southern Minnesota to test regional applicability of
the methods. In addition, about 10 wetlands moni-
tored by citizens will be assessed by MPCA using
the technical ffils. The data from the assessments
by citizens using the modified IBIs and MPCA's
technical ffils will be compared. The IBI results
will be compared with results from a Minnesota
rapid assessment method carried out by consult-
ants on the same 10 wetlands.
STUDY DESIGN
In the 1992 project on reference wetlands, 32
least disturbed and 3 known disturbed wetlands
were sampled for macroinvertebrates. In the sec-
ond research phase in 1995, 27 wetlands were
sampled for macroinvertebrates and vegetation.
The 27 wetlands represented the full range of hu-
man disturbance typical of wetlands in this part of
Minnesota, and were located in the North Central
Hardwood Forest ecoregion (Figure 9). The sites
included 6 least impaired reference sites and 21 sites
highly impacted from human disturbances such as
stormwater (12 sites) and agricultural influences (9
sites).
In the 1999 large depressional wetlands proj ect,
44 sites were sampled, 6 of which were sampled
twice on the same date. Included were 14 high-
quality reference sites, 14 agriculture-influenced
sites, and 16 urban wetlands (Figure 10). These
wetlands were selected to represent the widest range
of disturbance.
38
-------
1995 Wetland Site Locations
Reference Site
A Storm Water Site
Agricultural Site
^
FIGURE 9: LOCATIONS OF SMALLER DEPRESSIONAL WETLANDS
SAMPLED IN 1995.
Bco_
Lrgdepsites,shp
x Ag
« Ref
FIGURE TO: LOCATIONS OF THE 44
LARGE DEPRESSIONAL WETLANDS
SAMPLED IN 1999.
In the 2001 statistical assessment of wetlands
methods project, three sectors of nine wetlands
were sampled for invertebrates and vegetation.
In 2002, approximately 50 large depressional
wetlands representing a gradient of human influence
are being assessed. Approximately 30 wetlands in
the Western Cornbelt Plains and the Northern Gla-
ciated Plains ecoregions are being analyzed to de-
termine if the IBI developed in the North Central
Hardwood Forest (NCHF) ecoregion can be used
in other regions of Minnesota. Five of these sites
will have duplicated sampling. Ten wetlands from
previous proj ects in the NCHF ecoregion will be
analyzed. Another 10 wetlands in the NCHF
ecoregion are being sampled by citizens using the
IBIs and by consultants using a rapid assessment
method.
In the projects in 1995, 1999, 2001, and 2002,
both invertebrate and vegetation sampling were
done. Water chemistry samples were taken in June,
39
-------
and sediments were cored for metals and other fac-
tors in later summer.
In addition to invertebrate and vegetation sam-
pling, wetlands were sampled for water pH, con-
ductance, turbidity, dissolved oxygen, temperature,
calcium (hardness), chloride, total suspended sol-
ids, total phosphorus, and total nitrogen. Sediments
were analyzed for 15 heavy metals using ICP meth-
ods, as well as for total organic content, textural
classes, carbonates, chloride, total phosphorus, and
total nitrogen. Low-altitude aerial photographs were
taken to support the scoring for the human distur-
bance gradient.
SAMPLING METHODS: GENERAL
CONSIDERATIONS
Stratification of the habitat for each study site was
done so as to minimize the biological variability
among the different strata of the wetland. For the
depress!onal wetland projects, the following habi-
tats were identified: nearshore emergent zone, open
water submergent zone, and floating plant zone.
SAMPLING METHODS:
MACROINVERTEBRATES
Sampling was done in the nearshore emergent zone
during the seasonal index period of June to early
July. This time frame ensured that optimal species
maturity and richness were present. In previous
fieldwork, MPCA had determined that sampling in
May was too early because some invertebrates are
too immature for identification. Once collected, in-
vertebrate samples were preserved and analyzed
in the laboratory. Macroinvertebrates were sampled
using both the dipnet and the activity trap method.
Dipnetting captured the greatest richness of inver-
tebrates and the activity trap captured the active
swimmers and night-active predators.
Dipnet
The D-frame aquatic dipnet with 600-micron mesh
net was used. Two dipnet samples were taken within
the emergent vegetation zones. A %" wire screen
fixed to a wooden frame was used to keep the veg-
etation from the sample. After sweeping the net
strongly through the water column four to five times
and downwards to near the bottom, the contents of
the dipnet are emptied onto the framed wire screen.
The frame is set over a pan containing sieved water
to catch invertebrates as they drop down from the
vegetation. For approximately 10 minutes, the veg-
etation is gently spread and invertebrates are en-
couraged to drop or crawl down to the water in the
pan. After separation from the vegetation, the wa-
ter is then poured through a 200-micron sieve to
concentrate the sample before preservation. Pres-
ervation of the samples is done using 80% ethanol,
final concentration. Using a squirt bottle with the
alcohol solution, the sample is back-flushed into the
sample j ar and labeled for later picking and identi-
fication.
Activity trap
The bottle trap works as a passive funnel trap that
collects organisms as they swim into the funnel and
pass through the neck into the bottle. Made from a
clear 2-L round-bottomed plastic beverage bottle,
the traps are nearly invisible underwater. The traps
are supported on a 4.5' wooden dowel with a flex-
ible half section thin wall PVC pipe that allows rais-
ing and lowering the bottle trap on the dowel.
Ten bottle traps were placed in the emergent veg-
etation zone and left overnight for two consecutive
nights. Placement of the bottle traps was from the
nearest shallow shore edge to the inner side of the
deeper emergent vegetation zone in water no greater
than 1 meter. In the shallowest water, the traps
were piaced on the bottom just under the surface
of the water. Traps were placed horizontally about
15-20 cm under the surface. The bottle traps were
back-filled with water leaving no air bubbles inside
to reduce the amount of predation within the trap.
4O
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ANALYTICAL METHODS:
MACRO-INVERTEBRATES
After the invertebrates were sorted in the lab, the
entire sample was identified and counted. The data
were entered into an ACCESS database and
scored for 10 metrics, which represent different
measures of the invertebrate community. The metrics
used for small depressions are in the 1999 report
of the 1995 project. The metrics used for large
depressions are in the 2002 report on the 1999
project (Table 1). For the 1995 data, the metrics
were validated by plotting them against a ranking of
the site disturbance based on professional judgment
and against selected chemical variables. For 1999,
the metric data were plotted against the human dis-
turbance gradient scores (FIDS) and some chemi-
cal factors to evaluate metric responses. In addi-
tion, linear regressions were done on metric data
and ffils against the measures of human disturbance.
Chemical (e.g., phosphorus, nitrogen, chloride,
and heavy metals) and biological data were ana-
lyzed for statistically significant relationships to the
metrics and IBIs. Of the 10 invertebrate metrics,
intolerant taxa, chironomid taxa, and total taxa
showed the strongest responses to the estimated
disturbance gradient and water chemistry factors,
followed by the Odonata and ETSD metrics. HDS
scores, turbidity, and phosphorus and chloride in
water were most significantly related to the inverte-
brate IBI; copper in sediments was significant.
SAMPLING METHODS: VEGETATION
Vegetation sampling techniques vary greatly for dif-
ferent wetland habitats and study designs. MPCA
used a releve method for sampling vegetation. The
releve method was chosen for several reasons. The
primary reason is that it is easily adapted to widely
varying habitats and vegetation community struc-
ture, which is typical for depressional wetlands. This
adaptability in sampling methodology is needed for
wetlands that receive significant quantities of water
during storm events. A second reason that MPCA
selected releve sampling over line transect or quadrat
sampling was that the Minnesota Department of
Natural Resources' Natural Heritage Program and
County Biological Survey use releves, and
Minnesota's academic researchers also use a simi-
lar releve method for collecting vegetation commu-
nity data.
All vegetation sampling was done in July, which
represents the typical period of maturity and the
best time in Minnesota for determining community
structure. In each wetland, a 100-m2 plot was es-
tablished in representative sampling locations within
the emergent and open water submergent zones.
After establishing the releve plot, the vegetation
cover classes were determined for each plant spe-
cies occurring in the plot. Voucher specimens were
collected at least once during the proj ect for each
species or taxon encountered. All taxa that couldn't
be identified reliably to the species level in the field
were also collected.
ANALYTICAL METHODS: VEGETATION
Ten vegetation metrics were developed and vali-
dated, using methods similar to those for the inver-
tebrate IBI. Each promising attribute of the plant
community was plotted against a disturbance gra-
dient. In the 1995 small depressional wetlands
proj ect, the disturbance gradient index was devel-
oped from professional judgment ratings of several
disturbance factors including stormwater input, ag-
ricultural practices, quality of adjacent buffers, hy-
drologic alterations, and historical disturbances. For
the 1999 large depressional wetlands project, the
HDS scores and chemical factors were used. Metric
scoring criteria were then developed for the stron-
gest responding metrics (Table 2). Metrics were
also plotted against chemical variables to demon-
strate their response to traditional water chemistry
concerns.
41
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TABLE 1: SCORING CRITERIA FOR 1O INVERTEBRATE METRICS FOR IBI FOR
LARGE DEPRESSIONAL WETLANDS
METRIC DESCRIPTION
Number oi intolerant laxa: Leucorrliinia, Libellula, Tanylarsus. Procladius,
Triaenodes. Oecelis
MTSI) metric: £ genera mayflies, caddis flies: presence of fingernail clams
iSphaenidae) and dragonfly larvae
Tolerant laxa proportion of count of individuals to total sample count:
Tricliochorixa. hnallagma. hrpobdelk). Phvsa, C'rieotopus. Dicroiendipes.
Kndoehironomus. Glvpotendipes. and Paraumviarsus
Dominant three laxa: proportion of count of individuals in the dominant three laxa
to the total sample count
Corixidac proportion of beetles and hugs in acii\ itv trap samples: count of
individual Conxidac to total beetle and hug count
Chironomid uenera
Total number ol 'dragonfly and dumscllly genera (Odonata metric)
Leech laxa
Snail laxa
Tola! number of invertebrate laxa: larval ehironomids. caddisllies, mayflies.
dragonflies, damselflies, beetles, bugs, leeches, snails, niacrocrustaceans. dipk'ia
and fingernail clams
Foial possible scoring range for invertebrate IBI
Ranae for excellent condition
Raimc tor moderate condition
Range for poor condition
RANGE
5-1
3-4
0-2
7-10
4-6
0-3
<42%
>42-64IJ,,
69",,
- :54%
>54-74°u
>74 %
'o
33-670--,,
>67°,,
1 4 ->2 1
7-13
0-6
6-10
4-5
0-3
5-9
3-4
0-2
7-9
4-6
0-3
>52-77
37-52
01-36
SCORE
5
*
_>
i
s
3
1
^
3
1
5
t
_t
1
s
_>
1
_s
_i
1
S
5
i
5
j>
i
s
~t
_>
i
5
_i
1
1 0-50
35-50
23-34
1 0-22
The 10 vegetation metrics showed significant re-
sponses to water (chloride and phosphorus) and
sediment chemistry (zinc, copper, and nickel). Sen-
sitive species were found to be the strongest and
most reliable vegetation metrics.
SAMPLING METHODS:
WATER AND SEDIMENT
Sampling water and sediment chemistry was con-
ducted by MPCA staff. Water analysis was done
by the Minnesota Department of Health, and sedi-
ment analysis was done under contract with the
University of Minnesota Soils Analytical Labora-
tory. See April 1999 report for sampling methods
for sediment and water chemistry.
LESSONS LEARNED
Vegetation IBIs show great promise for future
applications in wetlands biological assessment.
The wetland plant community is biologically rich
and sensitive to a variety of human disturbances.
Data are acquired in a short time.
42
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TABLE 2: SCORING CRITERIA FOR 1O VEGETATION METRICS FOR
DEPRESSIONAL WETLANDS
LARGE
METRIC
Vas genera
Nonvas taxa
Carex co\ er
Sensitive species
Tolerant taxa
Grass-like laxa
Perennials
Aquatic guild
Proportion of
dominant 3 taxa
cover class
Persistent litter
METRIC DESCRIPTION
Number of vascular genera in the sample
Number of rtunvascular taxa
Sum of all Carex species combined cover
Taxa whose presence decreased with disturbance
Proportion of tolerant taxa to total taxa
Number of grass, rush, and sedge species
Count of all perennial species in the sample
Number of aquatic guild species
l-A|iiitabilit\ of perennial cover within the sample
Relative cover class (sum of individual cover class/total
sample cover class | taxa with persistent litter
RANGE
-- 1 4
8-14
<8
1
0
0.5-2
>4
2-4
<2
25.<>8-44.!S
-44.8
>6
3-6
-3
6-17
<6
5-8
<"5
(I So hS
>U.6S
17.3-34.6%
>34,6%
SCORE
5
1
5
1
s
Jl
1
s
1
S
\
i
5
1
S
1
5
1
5
1
5
1
The invertebrate community is sensitive to many
disturbances in wetlands. Responses differ from
stream invertebrates because wetlands inver-
tebrates are adapted to daily cycling of dissolved
oxygen in wetlands. What may be a "pollution
tolerant" invertebrate in streams may be a spe-
cialist in the wetland habitat.
Biological metrics respond to many stressors
and to other disturbance factors in the land-
scape and in physical aspects of the wetlands
that can be readily measured.
After applying appropriate stratifications, we
find that wetlands are not chaotic or highly vari-
able. They show clear patterns and predict-
able responses to human disturbances.
43
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MINNESOTA: DAKOTA COUNTY WETLAND HEALTH
EVALUATION PROJECT
Contact Daniel Huff
Environmental Education Coordinator
Organization Dakota County Environmental Education Program
4100 220th St. West, Suite 101
Farmington, MN 55024-9539
Phone (651)480-7734
E-mail daniel.huff@co.dakota.mn.us
Website http://www.extension.umn.edu/county/dakota/Environmentwetiands/wetid.html
PURPOSE
Evaluate wetland health using biological data
gathered by citizen volunteers using approved
techniques developed by the Minnesota Pollu-
tion Control Agency
Increase biodiversity in wetlands in urban ar-
eas by installing Best Management Practices
(BMPs)
Conduct a public wetland education effort
through seminars, workshops, field days, and
media
WETLAND TYPE
Depressional wetlands
ASSEMBLAGES
Macroinvertebrates
Vascular plants
STATUS
Since 1997, the project has operated every sum-
mer. In 2002, the program consisted of 15 moni-
toring teams representing 19 communities in Da-
kota and Hennepin Counties.
PROJECT DESCRIPTION
The Dakota County Wetland Health Evaluation
Proj ect (WHEP) uses sampling methods and evalu-
ation metrics developed by the Minnesota Pollu-
tion Control Agency (MFC A). The proj ect started
in 1997 and was conducted by MFC A and Minne-
sota Audubon. Atotal of 30 wetlands were moni-
tored by 5 citizen teams representing the Minne-
sota Zoo, Dakota County, and the cities of
Burnsville, Eagan, and Lakeville. MPCA staff
trained volunteers in sampling protocols, quality
assurance, and plant and macroinvertebrate identi-
fication. The sampling methods are a scaled-back
version of the Minnesota Pollution Control Agency
Wetland Bioassessment Program (see "Minnesota:
Developing Wetland Biocriteria" in this module).
Each citizen team worked under the direction of a
local teacher or nature center staff. The time com-
mitment for volunteers was approximately between
40-50 hours per year, including training, field work,
and analysis.
44
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In 1998, the project expanded to eight citizen
teams, and a technical consultant was hired to con-
duct MFC A's full sampling methods to facilitate
comparisons. Results of the comparison indicate
that the volunteer assessments, although not as rig-
orous as the professional assessments, provide re-
peatable results that are consistent with the more
detailed assessments. However, the volunteers tend
to score the high-quality wetlands too low because
they are unable to identify as many organisms (e.g.,
species of Carex) resulting in lower scores for the
richness metrics. A detailed description of this
proj ect is provided on the web site listed under the
contact information (http://www.extension.umn.edu/
county/dakota/Environment/wetlands/wetld.html).
In 1999, the funding source changed from EPA
grants to funding from the Minnesota State Legisla-
ture through the Minnesota Environment and Natu-
ral Resources Trust Fund. The Minnesota State
Legislature approved continued funding for this
project for the period of 1999 to June 2002. For
the 1999 season, the proj ect expanded to a total of
10 cities. An additional 11 wetlands were assessed
for the first time and 24 previously sampled wet-
lands were resampled for trend analysis. The in-
vertebrate and vegetation IB I scores were gener-
ally consistent for each wetland, although the inver-
tebrate score was slightly lower on average than
the vegetation scores. Each team performed a cross-
heck on a wetland monitored by another team. In
addition, the technical consultant field checked 4 of
the 3 5 wetlands sampled by the volunteer teams.
The volunteer results were compared to MFC A
standard sampling method results and were consis-
tent for most samples.
In 2000, 10 city teams monitored 38 wetlands.
Invertebrate scores were generally lower than veg-
etation scores, a more significant difference than the
previous year. Below-normal precipitation for the
previous fall, winter, and spring may account for
the poor showing of invertebrate populations. Vol-
unteers continued to use MPCA protocols and be
trained by the MPCA scientists who developed the
protocols. Eight of the 10 teams performed a cross-
check on a wetland monitored by another team.
Of these, six of the eight sites showed consistent
scores for the invertebrate index and seven of the
eight sites showed consistent scores for the vegeta-
tion index. The technical consultant field-checked
4 of the 3 8 wetlands sampled by the volunteer teams.
In general, the citizen data were consistent with
consultants' findings.
In 2001, 10 teams representing 11 cities moni-
tored 41 wetlands. A wet spring may have contrib-
uted to higher invertebrate scores than the 2000
monitoring season. Vegetation scores were con-
sistent with previous seasons' scores. Twenty-eight
of 39 wetlands sampled for both vegetation and
invertebrates were considered to have consistent
scores between the 2 indexes. Seven city cross-
checks were performed using the invertebrate
metrics. Of these seven, five resulted in similar
scores. Six city cross-checks were performed us-
ing the vegetation metrics. Of these six, four re-
sulted in similar scores. The technical consultant
field-checked 3 of the 41 wetlands sampled by the
volunteer teams. The consultant's check resulted
in identical scores for two of the three volunteer
teams and was very similar for the third for the in-
vertebrate metrics and the vegetation metrics. This
showed higher consistency between the citizen teams
and the professional cross-check than in previous
years. In addition to the 10 Dakota County teams,
3 teams within Hennepin County monitored 14
wetlands with funding from U.S. EPA and the
Minnehaha Creek Watershed District.
In addition to wetland monitoring, a wetland
remediation proj ect was begun in 2001 at one of
the wetlands previously monitored by the project.
Cedar Pond in Eagan, MN, had scored among the
lowest of all county wetlands for both vegetation
and invertebrates when sampled by volunteers in
2000. In cooperation with the City of Eagan, a
retaining wall surrounding the pond was removed,
the slope was regarded, and three zones of native
wetland vegetation, emergent, wet meadow, and
45
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upland buffer, were planted around the wetland.
Three rainwater gardens were constructed and
planted to receive and filter runoff from an adj acent
city street. WHEP volunteers participated in plant-
ing the rainwater gardens and trees along the up-
land buffer.
Complete reports of the 1999, 2000, and 2001
monitoring season results can be found on the web
site given above. Information on the 2001 moni-
toring season within Hennepin County can be found
at http://www.hcd.hennepin.mn.us/whep.html.
In 2002, communities sponsoring teams are pro-
viding funding for the proj ect. Nine teams repre-
senting 11 cities in Dakota County and 6 teams rep-
resenting 4 cities and 1 watershed district in
Hennepin County are participating in the Project.
Results from the 2002 monitoring season for both
counties will be available on the web in February
2003. The proj ect is expected to continue.
46
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Contact
MINNESOTA: UNIVERSITY OF MINNESOTA'S
IBI DEVELOPMENT
Sue Galatowitsch
Organization University of Minnesota
Department of Horticultural Science
3 05 Alderman Hall, 1970FolwellAve.
St. Paul, MN 55108
Phone (612) 624-3242
E-mail galat001@maroon.tc.umn.edu
Website http://www.hort.agri.umn.edu/mnwet/begin.htm
PURPOSE
Develop assessment methods to evaluate ecologi-
cal condition of wetlands
Variety
WETLAND TYPE
ASSEMBLAGES
Amphibians
Birds
Fish
Macroinvertebrates
Vascular plants
STATUS
Analyzing data and writing reports
PROJECT DESCRIPTION
Developing indices of biological integrity (ffils)
for Minnesota was pursued by researchers at the
University of Minnesota to enable quality assess-
ments of existing and restored wetlands. Eight
series of 15 wetlands (120 sites) were used to de-
velop wetland ffils. Each series covers a major
wetland type in the State and is composed of refer-
ence sites (unaltered wetlands in an unimpaired set-
ting), sites surrounded by land use typical of the
region, and sites that are highly altered. Plants, birds,
fish, invertebrates, and amphibians were surveyed
to select the best ffils for each series.
To identify possible patterns in biological com-
munities that may relate to land use differences, each
organismal data set (except amphibians-few organ-
isms encountered during surveys) was explored with
TWINSPAN. TWINSPAN organizes data so that
the most similar sites (as described by their spe-
cies) are grouped together as columns on a table,
and so that the species with similar habitat affinities
(as described by sites where they occur) are grouped
together as rows. This TWINSPAN table was used
to generate a list of potential indicators for analysis
with land-use data. Species or groups of species
that appeared preferential to sites with similar land
use characteristics were deemed to be potential in-
dicators. Other common ecological measures, such
as richness (number of species), were routinely in-
cluded in the list of potential indicators, as well.
Twenty-eight potential indicators identified for this
series are listed below. Proportional indicators for
animals are calculated as a total of all organisms
47
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observed (not a proportion of taxa) unless noted.
Absolute abundances for plant species (vegetation)
cannot be reliably estimated from cover class data.
Importance values, an approximate measure of
abundance, were calculated by summing cover class
scores (r=0.1,+=0.5, and classes 1-5).
Potential indicators
Amphibians: None
Birds (12): Species richness (BSR); total abun-
dance of all birds (B ABU); number of wetland taxa
(BWR); proportion of wetland birds (PWB); abun-
dance of brown-headed cowbirds (ABHC); pro-
portion of forest-nesting birds (PFB); number of
taxa with large territories (BLTR); proportion of
insectivorous birds (PBIN); abundance of marsh
and sedge wrens (CIS); abundance of yellow
throats, swamp sparrows, andLeConte's sparrows
(SSYT); number of open ground nesting species
(BOGR); proportion of open ground nesting spe-
cies (PBOG).
Fish: None
Invertebrates (4): Total abundance (IABU), taxa
richness (ISRI), number of snail species (GASR),
proportion of snails (PGAS).
Vegetation (12): Species richness (VSR), inva-
sive perennial species importance (TPI), number of
Carex species (CAR), importance of Carex spe-
cies (CAT), number of native herbaceous perenni-
als (HPNR), importance of native herbaceous pe-
rennials (HPNI), number of native perennial
graminoids (GPNR), importance of native peren-
nial graminoids (GPNI), number of introduced spe-
cies (INR), importance of introduced species (INI),
ratio of graminoids to herbaceous species (VGH),
ratio of annuals to perennials (VAP).
Relationship of potential indicators to land-
use measures
Values for each potential indicator (PI) were cal-
culated for each site in the series. PI values were
correlated with land use data: site alteration score,
land use cover at 500 m, 1,000 m, 2,500 m, and
5,000 m radii. For the radii data, six correlations
were calculated, one for each land cover category
(agriculture, urban, disturbed, forest, grassland, and
wetland). For this series a total of 700 relation-
ships were tested. Relationships with Pearson cor-
relation coefficients greater than 0.53 (p < 0.1) are
worthy of further consideration as indicators of
wetland quality. Each of these relationships were
plotted to detect if the high coefficients were based
on outliers. Those with outliers were not consid-
ered significant.
Eighteen of the original 28 potential indicators
(64%) were found to have a high correlation to land
use. Seven of the bird Pis show a land-use rela-
tionship. Birds with large territories (BLTR) are
more common to sites with less agriculture and dis-
turbance at the regional scale (2,500-5,000 m).
Likewise, regional patterns of urbanization are nega-
tively associated with wetland bird richness (2,500
m). Wetland bird richness is greater on sites with
more wetlands at this same scale. Overall bird rich-
ness is lower with more surrounding urbanization at
most scales. In contrast, most of the seven promis-
ing vegetation Pis show stronger land-use relation-
ships at local scales. Introduced species (INI, INR)
and annuals (VAP) are more common on sites that
are immediately impacted by agriculture and urban-
ization (site). Four invertebrate Pis will be further
considered for indicator development. The
richness of snail taxa (GASR) is positively associ-
ated with forest and wetland cover from 1,000 to
5,000m.
48
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MONTANA: DEVELOPING WETLAND BIOASSESSMENT
PROTOCOLS To SUPPORT AQUATIC LIFE BENEFICIAL
USE-SUPPORT DETERMINATIONS
Contact
Randall S. Apfelbeck
Organization Montana Department of Environmental Quality
2209 Phoenix Avenue
P.O. Box 200901
Helena, MT 59620-0901
Phone (406) 444-2709
E-mail rapfelbeck@state.mt.us
Website http://www.deq.state.mt. us/ppa/mdm/Wetlands/Lakes&Wetlands_Index.asp
PURPOSE
Montana's wetland research objectives are to:
Determine the status and trends in wetland wa-
ter quality.
Acquire an understanding of how climate, hy-
drologic controls, and geomorphic settings in-
fluence wetland biological communities for the
development of successful biocriteria.
Develop biological measurements that could be
used in developing biocriteria to define the ex-
tent and degree of anthropogenic impacts to
wetland water quality.
Develop an integrated assessment of wetland,
streams and lakes within a watershed.
Report on aquatic health at the watershed level
through the development of landscape assess-
ment tools.
Develop rapid field assessment protocols to
evaluate aquatic ecological conditions by using
indicators and best professional judgement.
WETLAND TYPE
Variety of wetland types
ASSEMBLAGES
Algae
Macroinvertebrates
Vegetation
Amphibians and aquatic reptiles
STATUS
Ongoing, revising analytical methods
PROJECT DESCRIPTIONS
1992
Montana Department of Environmental Quality
(DEQ) began developing wetland biological crite-
ria. At that time, there was little information con-
cerning the status or trends of the water quality of
Montana's wetlands. Furthermore, Montana's
water quality standards were developed to protect
the beneficial uses (e.g., aquatic life) of lakes; river
49
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and stream wetlands were not considered State
waters when Montana's water quality standards
were developed. Since 1992, Montana has had an
ongoing program to develop bioassessment proto-
cols and water quality standards that will more ad-
equately evaluate and protect the aquatic life that
live in wetlands.
In 1998, as a result of a TMDL lawsuit, Montana
DEQ shifted focus from development of biological
criteria to the development of guidelines for making
beneficial use decisions that apply to all Montana
waterbodies. Montana's guidelines for making ben-
eficial use decisions can be found at http://
nri s. state, mt.us/wis/tmdlapp/pdf 20 027
Appendix_A.pdf.
1998
In conjunction with Montana DEQ's research pro-
gram, Montana State University (MSU) designed
a study that focused on development of vegetation
biocriteria for western Montana depress!onal wet-
lands (Borth 1998). The focus on vegetation
biocriteria is key in Montana because wetland veg-
etation is easier to assess than macroinvertebrates
or diatoms for depressional wetlands that are sea-
sonally dry. The MSU study sampled vegetation
and also macroinvertebrates and diatoms for 24 de-
pressional wetlands with similar climate, hydrology,
and water chemistry. The research included sam-
pling across three levels of human disturbances
minimally impacted, slightly impacted, and moder-
ately impacted. The study also involved two an-
thropogenic impairmentsdryland agriculture and
grazing.
2000
Researchers from the University of Montana con-
ducted a study to determine the effects of natural
variability on the use of macroinvertebrates as
bioindicators of disturbance in intermontane depres-
sional wetlands in northwestern Montana (Ludden
2000). Their study design included collection of
macroinvertebrate samples andphysiochemical data
from 15 pristine and 6 disturbed intermontane prai-
rie potholes. Their study also included analysis of
macroinvertebrate samples that were previously
collected by Borth (1998). The researchers col-
lected macroinvertebrate samples from across three
seasons and from three wetland zones. They used
multivariate detrended correspondence analysis to
ordinate the raw macroinvertebrate data and
physiochemical variables as secondary matrices to
establish vectored biplots of correlation. Candi-
date metrics were analyzed using univariate analy-
sis. They determined that no environmental vari-
ables were strongly correlated with the
macroinvertebrate data, and many of the metrics
varied across wetland zones and across seasons.
Nevertheless, 35 candidate metrics were able to
discriminate between minimally and highly disturbed
sites. These wetlands were also sampled intensively
to develop and test a model for assessing depres-
sional wetlands using the hydrogeomorphic (HGM)
functional assessment approach.
2002
Montana DEQ initiated the development of a
comprehensive watershed monitoring and assess-
ment program. The comprehensive program will
be developed to determine the causes, effects, and
extent of pollution to aquatic resources (including
wetlands) and for developing pollution prevention,
reduction, and elimination strategies.
The comprehensive program has three compo-
nents: landscape, site-specific, and rapid assess-
ment. First, a landscape-level process to evaluate
and rank wetlands and watersheds for protection
and restoration is critical to maximize the use of lim-
ited financial and management resources. This pro-
cess will use existing digital data and evaluate a range
of landscape impacts using a Geographic Informa-
tion System. Second, Montana will continue to
develop site-specific assessment protocols such as
biocriteria (i.e., for assessing amphibians, vegeta-
tion, algae, and macroinvertebrate assemblages).
This information is important for managers of
Montana's water resources for making informed
watershed management decisions. Development
5O
-------
of site-specific wetland biological, physical, and
chemical assessment tools will be used to evaluate
aquatic life beneficial use support to determine im-
pairment as per section 303(d) of the Clean Water
Act. Third, like most States, Montana has very
limited resources for assessing wetland water qual-
ity and aquatic conditions for 305(b) purposes. For
this reason, Montana will develop an assessment
approach that is truly rapid and cost effective. A
rapid assessment component will be developed
based upon field data collected, tested and refined
during this study, best professional judgment and
information gathered from a literature search.
The comprehensive watershed monitoring and
assessment program will include a probabilistic study
design to assess the ecological condition of depres-
sional and riverine wetlands and streams. Three
pilot subbasins (fourth code HUCs) will be selected.
The pilot subbasins will represent Rocky Moun-
tain, Intermountain Valleys and Prairie Foothills, and
Plains ecoregions (Figure 11).
Within each pilot subbasin, three sixth-code FIUCs
(fifth-code HUCs for the Middle Milk subbasin)
will be randomly selected from each of three dis-
turbance strata (high, medium, and low) determined
from landscape-scale assessments. For vegetation,
soils, water chemistry, macroinvertebrates, diatoms,
and land use, a total of six wetlands (three riverine
and three depress!onal) will be monitored in each
watershed. A total of 27 watersheds (9 in each
subbasin) and 162 wetlands (3 riverine and three
depressional in each watershed) will be studied. For
the determination of the detection/nondetection of
amphibians and aquatic reptiles, all standing water
bodies identified in each watershed will be surveyed.
Note: the remainder of this case study will de-
scribe in more detail the study initiated in 1992.
STUDY DESIGN
The original Montana DEQ study was designed
in 1992 and involved sampling 80 wetlands
throughout Montana during 1993 and 1994. The
bioassessment project included collection of
macroinvertebrate and diatom samples from wet-
lands in all ecoregions of Montana (Apfelbeck
2001).
Montana DEQ's approach to developing
biocriteria involved several study designs aimed at
developing tools to help detect human influence on
wetland water quality. The original study was de-
signed in 1992 and involved sampling 80 wetlands
r 1 Pllol Wllmhnll
Ecorraions
Rts.li \Ioilnlaln\
MoMnu ViHty mi FMM1I Pr.lrlf*
Grrat Plains
\\ jotiiltiB BasLn
it V M
JMirt, Nnonbn 1», iOB!
FIGURE 11: MONTANA ECOREGIONS AND PILOT WATERSHEDS.
51
-------
throughout Montana during 1993 and 1994 (Fig-
ure 12). The study design included collection of
samples that represent the wetland's
macroinvertebrate (e.g., aquatic insects) and dia-
tom (algae) communities. Sampling methods were
designed so that 1 -2 hours was sufficient for the
data to be collected in the field for each wetland.
Samples of each wetland's water column, sediment,
and macroinvertebrate and diatom communities
were collected. Water-column and sediment
samples were collected to document impairments
and for classification purposes.
Ecoregions of Montana
50 100 Miles
I I Montana Valleys and Foothill Prairies
I I Montana Plains
Montana Rocky Mountains
A.L Gallant, U.S. EPA. 1987
FIGURE 12: ECOREGIONS AND SAMPLING LOCATIONS BY WETLAND CLASS.
Wetland Classes: (referred to by number in the above figure)
Class 1 Dilute Closed Basins and Headwater Wetlands of the Rocky Mountain Ecoregion
Class 2 Riparian Wetlands of the Rocky Mountain amd Intermountain Valley Ecoregions
Class 3 Groundwater Recharge Closed Basin Wetlands
Class 4 Riparian Wetlands of the Plains Ecoregion
Class 5 Alkaline Closed Basin Wetlands
Class 6 Saline Wetlands
Class 1 Surface Water Supported Closed Basin Wetlands
Class 8 Ephemeral Wetlands
Class 9 Open Lake Wetlands of the Plains Ecoregion
Class 10 Open Lake Wetlands of the Rocky Mountain and Intermountain Valley Ecoregions
52
-------
The sites were classified using ecoregions and
hydrogeomorphology, and several of the wetland
classes were further delineated using water-column
chemistry variables. A representative number of
wetlands from the following Omernik ecoregions
were sampled: Rocky Mountains, Intermountain
Valleys and Prairie Foothills, Glaciated Plains, and
Unglaciated Plains Ecoregions. To reduce season-
ality, all wetlands within the same ecoregion were
sampled during similar time periods. Wetlands in
the Glaciated Plains Ecoregion were sampled from
early April through mid-June, wetlands of the Inter-
mountain Valleys and Prairie Foothills Ecoregion from
mid-June until early August, and wetlands of the
Rocky Mountain Ecoregion from early July through
September.
The classification framework was developed by
sampling the full spectrum of wetland types in Mon-
tana. The study was designed such that approxi-
mately 75% of the sites were reference and 25%
were impaired. This approach was useful because
it allowed Montana DEQ to determine the refer-
ence condition of a wide variety of wetland types.
Also, the design provided the opportunity to test
the ability of the biological measurements to detect
water quality impairment.
Anthropogenic impacts such as irrigation or log-
ging were included in the study design. If anthro-
pogenic activities such as dryland agriculture, irri-
gation, feedlots, grazing, silviculture, road construc-
tion, hydrologic manipulation, urban runoff, waste-
water, mining, and oil and natural gas production
occurred in the wetland's watershed, the wetland
was considered impaired. Wetlands for sampling
were selected on the basis of many variables, in-
cluding availability of historical data, special inter-
ests by other entities, cooperation by landowners,
and accessibility.
In order to classify or document impairment, a
hydrogeologist for the Montana Natural Heritage
Program (MNHP) assisted Montana DEQ in de-
veloping a wetland classification system through
summarizing and interpreting the physical and chemi-
cal data. Using topographic maps, field observa-
tions, and information gathered from land manage-
ment agencies, geomorphic characteristics were in-
terpreted and a hydrogeomorphic database devel-
oped. Maps for each wetland were completed us-
ing a Geographic Information System (GIS). Map
features included hydrologic delineations, land man-
agement areas, counties, cities, major transporta-
tion corridors, wetland watershed boundaries, and
sampling locations. Visit Montana DEQ's website
to get more detail on the types of wetland classes
and for photographs of each type: http://
www.deq.state.mt.us/ppa/mdm/Wetlands/
classification.asp.
SAMPLING METHODS: DIATOMS
Montana DEQ collected diatoms as composite
grab samples. The algae were identified to the lowest
taxonomic level possible. Samples were collected
using a 250-mL plastic container and then preserved
with Lugol's solution. Samples were collected from
a location determined to best represent the wet-
land. These locations were restricted to areas that
were easily accessible when wearing hip boots.
Sampling was done from April through September.
Each site was sampled once.
ANALYTICAL METHODS: DIATOMS
The multivariate approach was used to analyze
wetland diatom communities. Multivariate analysis
is a statistical approach used by biologists to deter-
mine relationships among biota such as diatoms or
macroinvertebrates, and environmental variables
such as water-column chemistry. The multivariate
approach to investigate relationships between Mon-
tana wetland diatom assemblages and environmen-
tal variables (mostly water-column chemistry) was
detrended canonical correspondence analysis
(DCCA) and two-way indicator analysis
53
-------
(TWINSPAN). Clusters of diatoms with similar
composition were graphically displayed using
DCCA. Vectors were displayed and labeled to
illustrate the relationship between diatom assem-
blages and environmental variables. Longer vec-
tors show us a stronger correlation among diatom
assemblages and environmental variables (Figure
13). Envelopes were used to graphically enclose
all reference sites using the wetland class delinea-
tions (Figure 14). The Academy of Natural Sci-
ences of Philadelphia (ANSP) performed the
subsampling, digestion, and mounting of the dia-
toms (see Charles et al. 1996).
PHOSPHORUS.
LEAD
9
CONOUCTWrTY x *--^f ,^C«0 * +
»5JL^S£\I"X ' .
FIGURE 13: DIATOM DATA RELATED
TO ABIOTIC FACTORS USING
DETRENDED CANONICAL
CORRESPONDENCE ANALYSIS.
PHOSPHOROUS
V.JJ.
Aoata
0 '
WET 58
V t>
0 Cla&s I DArfp ClE-wil SQOHIS and
Welland* i>1 flw Rocky M
A Clftsa 6 Sdbrie Wetrtnda
4 Class 8 Epl-iempral WeBands
(a)
Phosphorous
\
Mel ah
*- WET 70
if
c
WEJ2
I
t.
m +
WET 78
t7^\*^Ei64
sf ^\ V *" DMule
*^r
A nlafec 7 QlMIWIhA UUAtlfl
(b)
Mountain and I hlerni'ou-'ilain Valley
9 Class 3 Qfiiuridwater RwdiafQi- Clustd Baa
Class 10 Op*fi Uk« W'iiiiiiH-|s, c,1 Qi« R.nckv
R«KitwK0* CwseH Bnain
X C-**.s 5 A».dW* CMiefl flnfcn Wetland*
Ohm 7 Swhn* Walw StwUtM Ch>w
-------
SAMPLING METHODS:
MACROINVERTEBRATES
Montana DEQ collected macroinvertebrates us-
ing a 1-mm mesh D-net in a sweeping motion.
Macroinvertebrates were collected from all microen-
vironments in a sampling location. These locations
were restricted to areas that were easily accessible
when wearing hip boots and that best represented
the wetland. Samples were composited with asso-
ciated materials such as vegetation and sediment
and then preserved with ethanol in a 1-L plastic
container. An effort was made to collect 300 or-
ganisms from each location using a consistent
method of collection. To ensure preservation,
sample bottles were refreshed with ethanol several
days after collection. Sampling was done once per
site from April through September.
ANALYTICAL METHODS:
MACROINVERTEBRATES
The multimetric approach was used to evaluate
wetland macroinvertebrate communities. The
multimetric approach incorporates many attributes
into the assessment process and has the ability to
integrate information from the biological communi-
ties to provide an overall indication of biological
condition or ecological health.
The proj ect contractor assisted Montana DEQ in
developing wetland macroinvertebrate multimetric
indices. The proposed metrics and associated en-
vironmental data were evaluated in an attempt to
develop an understanding of ecological relationships,
to test each proposed metric's ability to predict
various anthropogenic stressors, and to test redun-
dancy. Table 3 lists some of the proposed metrics
used for macroinvertebrates and Figure 15 shows
macroinvertebrate index values for several classes
ofwetlands.
The macroinvertebrate samples are subsampled
and sorted by contractors using a gridded sorting
pan. Asubsampling of 200 organisms is performed
for analysis. All individuals are counted when there
are less than 200 organisms in the sample. Organ-
isms are identified to the lowest taxonomic level (at
least genus if possible). The taxonomic level of iden-
tification is standardized using the Montana Stream
when possible. Amphipoda are identified by spe-
cies. Common, easily identified midge taxa are iden-
tified using a dissecting microscope equipped with
25x oculars. Midge taxa that require slide mount-
ing are cleaned with warm water solution of 10%
KOX, rinsed in distilled water followed by 95%
ethanol, and mounted on slides in Euperal. If fewer
than 30 individuals are to be mounted, all individu-
als are mounted. If there are more than 30 midge
larvae, at least 10% of each morphotype are
mounted. Mounted midges are identified using a
Zeiss Axiolab phase contrast compound microscope
or equivalent.
The contractor identified the organisms in the
wetland samples and standardized the taxonomic
level of identification based on Montana Stream
Protocols. Several taxa were eliminated from con-
sideration for metric development, as they were
determined to be nonbenthic taxa or semiaquatic
surface dwellers and considered uninformative for
reflecting water quality. These taxa included
Gerridae, Collembola, Dytiscidae, Hydrophilidae,
Ostracoda, Anostraca, Copepoda, Cladocera,
Notonectidae, and Corixidae.
OTHER PARAMETERS: WATER
CHEMISTRY AND SEDIMENT
Water-column and sediment samples were col-
lected to document impairments and for classifica-
tion purposes. Each sample was collected from a
location determined to best represent the wetland.
These locations were restricted to areas that were
easily accessible when wearing hip boots. Field
chemical measurements, observations, and photo-
graphs were recorded at each location.
-------
TABLE 3: PROPOSED METRICS, PROPOSED METRIC CALCULATIONS, AND SCORE
CALCULATIONS USED FOR DEVELOPING WETLAND MACROINVERTEBRATE INDICES
PROPOSED METRICS
Nu in her of tax a
Percent dominance
Percent 1 dominant tason
Percent 2 dominant ia.su
Percent 5 dominant lava
POET
Number of individuals
Chironomidae
Number ofchironomidae taxa
Percent ehironomidac uixa
Percent Ortliocladiinae/
Chironomidae
Crustacea/Mollusca
Number ol'enistaeea &
mollusea la\a
Percent crusiacea & mollusea
taxa
Leech/S pon gc/CIa m
THEORIZED
DIRECTION OF
CHANGE IN
PRESENCE OF
STRESSOR
[Jeer ease
Increase
In ere rise
Increase
Increase
Decrease
Decrease
Decrease
Decrease
Increase
Decrease
Decrease
Decrease
Increase
Decrease
PROPOSED
METRIC
CALCULATION
Count laxa
Average of percent 1, 2,
and 5 mosl dominanl taxa
Count Pleeoptera,
( kionata, liphemeropiera.
and Trieoplera taxa
Count individuals in total
sample (maximum count
of 300)
Number of Ohir taxa
( I0(l%ehir i- 50) -
(((% onhocladiinae to
total Chir) MM)) +0.5)
Number of crustaeea and
mollusca taxa ( 100%
cruslaeea/niollusea laxa <-
511)
Count i lirudinea.
Porilera, and Sphaeriidea
liixa
SCORE
CALCULATION
(number of taxa) 0.75
( lOll"-,, dominance) , 0.36
(POr.T) - 3
(number of individuals)
3 3
(Chironomidae) - 83
(erusUieea'mollusea) 33
( leech 'sponge 'clam i 3
LESSONS LEARNED
We found that diatoms and macroinvertebrates
were most useful for evaluating the biological
integrity of perennial wetlands with open-water
environments that had relatively stable water
levels and were not excessively alkaline or sa-
line.
We concluded that multivariate analysis was a
useful tool for developing a wetland classifica-
tion system and that hydrogeomorphology and
ecoregions were practical approaches to clas-
sifying wetlands for the development of
biocriteria.
We determined that both the multimetric and
multivariate techniques were valuable for de-
veloping wetland biocriteria.
In most cases, the multimetric and multivariate
approaches that we used to assess the
macroinvertebrate and diatom communities both
identified the same wetlands as impaired.
Two wetland types in the arid west (including
Montana) are difficult to classify. Wetlands such
as potholes are highly complex and difficult to
classify because of both spatial and temporal
variability. For these wetlands, the hydrology,
water chemistry, and biology can change dra-
56
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Class 1. Dilute closed basins and headwater wetlands of the Rocky Mountain Ecoregion
C] LEECH'SSONCEICLAU
IB CRUSTACEA J MOUU3CA
O CHIRQHCMIDAE (MlOGEj
Q I OF INDIVIDUALS
Q PLEC/OOON'EFH/TRIC
CZ] INVERSE K DOMINANCE
I OF TAKA
Class 8. Ephemeral wetlands
METRICS
Q LEECH'SPONGE/CLAM
03 CRUSTACEA!MOLLUSCA
Q CHIRONOMIDAE IMIDG6
O « Of INDIVIDUALS
O FLEC/ODOH/EPHITRIC
Q INVERSE '-. DOMINANCE
f Of TAXA
Class 10. Open lake wetlands of the Rocky Mountain and Intermountain Valley ecoregions
£3 LEECH/SPOUSE(CLAM
CRUSTACEA IMOLLUSCA
CD CHIRONOMiOAE lUIDGG
Q « OF INDIVIDUALS
O PL6C/ODON(EPM/TSIC
El INVERSES DOMINANCE
TAXA
flt'li »ETIH *£Tti
YIEHS ftETSI SETT! Vl£;7- »ET7j »t:v; '.V E"
WETLAND CODE NAVES
FIGURE 15: MACROINVERTEBRATE INDEX SCORES FOR WETLANDS
IN SEVERAL CLASSES.
57
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matically throughout a season or from year to
year as a result of climatic change. For example,
the biological community of a wetland often
changes through an increase in salinity or a de-
crease in water content caused by drought.
Aquatic macroinvertebrate and diatom
biocriteria did not appear to be very useful for
detecting impairment in wetlands that usually
lack surface water. Vegetation biocriteria are
likely to be the most appropriate for assessing
the biological conditions of these wetland types.
A more quantitative approach i s needed for the
measurement of wetland physical disturbances
if we are to link physical disturbances to
changes in biological communities.
Note: Montana DEQ has developed a set of pro-
posed metrics, proposed metrics calculations, and
score calculations used for developing wetland
macroinvertebrate indices. These are included in
Tables.
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NORTH DAKOTA: WETLAND BIOASSESSMENT
PROTOCOLS FOR MAKING AQUATIC LIFE BENEFICIAL
USE-SUPPORT DETERMINATIONS
Contact
Mike Ell
Organization North Dakota Department of Health
Division of Water Quality
1200 Missouri Avenue
P.O. Box 5520
Bismarck, ND 04333
Phone (701)328-5214
E-mail mell@state.nd.us
PURPOSE
The primary purpose of North Dakota's wetland
bioassessment program is to develop wetland wa-
ter quality standards for North Dakota. This cur-
rently involves developing biological community
metrics and an Index of Biological Integrity (IB I)
for temporary and seasonal depress!onal wetlands.
A secondary project goal is to compare the IBI
with hydrogeomorphic (HGM) functional assess-
ments.
WETLAND TYPE
Depressional wetlands
ASSEMBLAGES
Algae
Macroinvertebrates
Vascular plants
STATUS
Analyzing data, developing IBIs, and expand-
ing project
PROJECT DESCRIPTION
North Dakota Department of Health (NDDH)
initiated its wetland bioassessment program in 1993
as a component of the State's strategy to develop
wetland water quality standards. NDDH began field
sampling in 1995 and is currently analyzing data
collected from 1995 through 1999 to develop as-
semblage metrics, IBIs for the assemblages, and
ultimately biocriteria. The NDDH wetland sam-
pling plan includes vascular plants,
macroinvertebrates, and algal assemblages.
Development of a North Dakota sampling
protocol for the State's wetland types is ongoing
through cooperative work agreements with North
Dakota State University, Departments of Animal
and Range Sciences and Zoology.
STUDY DESIGN
North Dakota's study design has continued to
expand in scope since the proj ect was initiated. Ini-
tial field sampling began in 1995 and 1996 with 13
"least disturbed" temporary and seasonal wetlands.
In 1997, the number and range of wetlands was
expanded to 20 temporary and seasonal wetlands.
Four of the original 13 "reference" wetlands sampled
59
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in 1995 and 1996 were sampled in 1997 along with
16 new wetlands. These wetlands captured a larger
range of disturbance categories.
In 1998, the number of wetlands again expanded.
An additional 16 temporary and seasonal wetlands
were added, bringing the total number of wetlands
sampled to 3 6 in 1998 and the total number of ref-
erence wetlands to 6. The 16 wetlands added in
1998 were part of an HGM classification project.
These wetlands were added to facilitate a compari-
son of the two assessment methods. In addition to
the 16 HGM wetlands, NRCS personnel conducted
an HGM assessment for the 20 wetlands selected
by the Department.
During the 1999 sampling season, a second set of
30 temporary and seasonal depress!onal wetlands,
not sampled previously, were sampled for vascular
plants. Macroinvertebrates and algae were not
sampled in 1999.
Since the inception of the project, temporary and
seasonal wetlands have been the focus of IB I de-
velopment efforts. Further, all wetland study sites
have been within the Northern Glaciated Plains and
Northwestern Glaciated Plains ecoregions (Figure
16), a region commonly referred to as the "Prairie
Pothole Region."
SAMPLING METHODS:
MACROINVERTEBRATES
NDDH currently uses only the "sweep" or jab
net method for macroinvertebrate sampling; how-
ever, in the past, activity traps have also been used.
Sampling consists of two site visits in June. During
each site visit two samples are collected with the
emergent vegetative zone.
Sweep net
The method employs a "D" frame net (0.6-mm
mesh size). At each site within the wetland, the
substrate (i.e., wetland vegetation and benthos) is
"jabbed" for a distance of 1 m. The disturbed area
is then swept two more times to ensure a represen-
tative sample of macroinvertebrates is collected.
Each sample collected within the wetland will be
placed in a shallow pan where any excess debris or
water can be removed. The "cleaned" sample is
placed in a j ar a preserved with buffered formalin
to a concentration of 10% by volume.
ANALYTICAL METHODS:
MACROINVERTEBRATES
In the laboratory each sample is washed through
a 0.6-mm sieve to remove the preservative and
cleaned to remove excess debris. The cleaned
sample is placed in a shallow white pan divided in
quadrants of equal size. Because each sample typi-
cally contains a large number of organisms, it is nec-
essary to subsample. A subsample is obtained by
randomly selecting a sample quadrant and counting
and identifying the organisms in that quadrant. Ad-
ditional quadrants are selected at random and or-
ganisms counted and identified until a subsample of
300 organisms is counted. For samples with less
than 300 organisms, every quadrant is sampled. All
organisms are identified to the lowest taxonomic
level practical.
SAMPLING METHODS: ALGAE
(PHYTO PLANKTON)
Phytoplankton samples are collected just below
the water's surface in the middle or deepest area of
the wetland basin. Each wetland is sampled two
times in June at the same time macroinvertebrate
samples are collected. The sample is collected by
submerging a clean 200-mL sample bottle just be-
low the water's surface and filling it. The sample is
preserved in the field with M3 fixative to a concen-
tration of 2% by volume.
6O
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17 Middle Rockies
I I ra Black Hills 1-oothilN
I I Pb Black Mills Plateau
I I PC Black Hills Core Highland*
25 \\ i-Mri ii I h^h IM.IIMS
I I 25a Pine Ridge L'scarpnmil
42 Northwestern Glaciated Plains
I I 42a Missouri C'oieau
I I 421) C'ollapsrd Glacial Outwaah
I I 42c .Missouri t'oteau Slope
I | 42d Northern Missouri Cotemi
| 42r .Southern Missouri Coteau
| 42fSoutheni Missouri Cou-au Slope
I 42 g Pouca Plains
I I 42h Southern River Breaks
I I 42i Glaciated Dark Bnmn Prairie
43 Northwestern (treat Plains
I I 43a Missouri Plateau
I I 43b Little Missouri Badlands
I I 43c River Breaks
| 43d Forested Dulles
| 43e Sagebrush Steppe
I | Hi sit him in ui Pierre Shale Plains
| 43g Semiarid Pierre Shale Plains
I 43h While River Badlands
| 43i Keva I'.ih.i Tablelands
I | 43j Vtoreati Prairie
| 43k Dense Clay Prairie
44 Nebraska Sand Hills
I I 44a Nebraska Sand Hills
46 Northern Glaciated Plains
|^H lf';l- Pembina Kscai~pmrnl
^^gj 46b Turtle Mountains
I I 46c Glacial Lake Basins
4(ul Glacial Uke Deltas
46e Tewaukon Dead Ice Moraine
46f End Moraine Complex
46g Vorihern Black Prairie
46h Noilheiit Dark Brown Prairie
46i Drift Plains
46j (il.ui.il Oulwash
46k Prairie Coieau
461 Prairie Coteau Escarpment
46m Big Sioux Basin
| | 46u James River Lonland
I | 46o Minnesota Ri>eij Prairie
1" Western C'orn Bell Plains
| | 4^a Loess Prairies
| ] 4^d Missouri Alluvial Plain
48 Lake Agassi; Plain
I | 4«a Glacial Lake Agassiz Basin
| | 4Sb Sand Delias and Beach Ridges
I I 48c Saline Area
FIGURE 16: LEVEL IV ECOREGIONS OF NORTH DAKOTA.
61
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ANALYTICAL METHODS: ALGAE
(P H YTO P LAN KTO N)
Identification and enumeration of phytoplankton
is made using the phase-contrast inverted micro-
scope method (Utermohl 1958). Sample analysis
is conducted from a sample aliquot ranging from 2
to 15 mL, depending on turbidity. Phytoplankton
greater than 20 micrometers in diameter are enu-
merated at x 125 magnification. (Note: At x 125
the entire bottom of the chamber is in view.) Fol-
lowing enumeration at x!25 smaller algae are
counted at x 1,250. At least 250 cells of the most
numerous algae are counted using the strip count
method at x 1,250 magnification (APHA 1989).
Cell volumes are estimated for dominant taxa by
measuring cell dimensions of 50 to 100 individuals
using the closest geometric formulas of Wetzel and
Likens (1991) andTikkanen (1986). For rare taxa,
volume estimates are made from fewer than 50 cell
measurements. Diatoms will be identified after
clearing in 30% hydrogen peroxide and mounting
in Hyrax Mounting Medium.
SAMPLING METHODS:
VASCULAR PLANTS
Working in cooperation with NDSU, the Depart-
ment is evaluating three methods to sample the vas-
cular plant community. The first method is more
qualitative in nature. It involves a simple inventory
of plant species present within each wetland zone,
low prairie, wet meadow, and shallow marsh. The
other two methods, termed the point and quadrat
methods, are quantitative. The inventory is con-
ducted each time the wetland is visited. The point
and quadrat methods are done once each, usually
in July or August.
In the point method, 200 points are evenly strati-
fied in each wetland zone and the nearest plant spe-
cies recorded. The quadrat method involves plac-
ing 15 1 -m2 quadrats evenly throughout each wet-
land zone. Each species is recorded within the
quadrat and a Daubenmire cover class is recorded.
With both methods, a secondary species list is made
for species encountered, but not sampled.
Note: For specific details on abiotic sampling
methods for nutrients, trace elements, general wa-
ter parameters, and sediment chemistry, directly refer
to North Dakota Department of Health.
LESSONS LEARNED
NDDH has made a great deal of progress within
the past 5 years. Our experience, however, has
not been without problems and mistakes. As men-
tioned previously, the first 2 years of the program
focused solely on "least impaired" or reference con-
dition wetlands. Although beneficial in testing sam-
pling methods, the lack of a disturbance gradient in
the study design did allow for the testing of attributes
and the selection of metrics. Therefore, beginning
in 1997 the NDDH chose wetlands across a dis-
turbance gradient, including both reference wetlands
and degraded wetlands.
NDDH has also found it beneficial to stratify wet-
lands based on ecoregion and wetland class. This
minimizes the amount of variation in the biological
assemblage and allows more sensitivity in the re-
sponse of the metrics to the disturbance gradient.
Current IBI development efforts are focusing on
temporary and seasonal depressional wetlands
within the Northern Glaciated Plains and North-
western Glaciated Plains ecoregions. In 2000 the
NDDH will be cooperating in two proj ects to de-
velop wetland IBIs for semipermanent depressional
wetlands and for floodplain wetlands along the Mis-
souri River.
62
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OHIO: DEVELOPING IBI ASSESSMENT METHODS
FOR WATER QUALITY STANDARDS AND
REGULATORY DECISIONS
Contact
John J. Mack
Organization Ohio EPA
Division of Surface Water
122 South Front Street
P.O. Box 1049
Columbus, OH 43216-1049
Phone (614) 644-3076
E-mail john.mack@epa.state.oh.us
PURPOSE
Test and develop biological criteria for wetlands
using vascular plants, macroinvertebrates, and
amphibians as indices of biological integrity
(IBIs) for eventual adoption into the State's
water quality standards
Use results from IBIs to calibrate the Ohio Rapid
Assessment Method for Wetlands to support
regulatory decisionmaking under the State's
wetland antidegradation rule, which requires that
wetlands be assigned to one of three catego-
ries based on the wetland's quality and func-
tionality.
WETLAND TYPE
Depressional wetlands
ASSEMBLAGES
Algae
Macroinvertebrates
Vascular plants
STATUS
Refining metrics and developing IBIs
PROJECT DESCRIPTION
The initial objective of this study is to provide the
reference data needed to implement the wetland
water quality standards and wetland antidegradation
rule. The pilot metrics developed from this study
should enable Ohio wetlands to be assigned to one
of the three regulatory categories. Generally, the
study objectives are as follows:
1. Develop pilot biological metrics that may be
used to evaluate the function and ecological in-
tegrity of a wetland. These metrics will be based
on the vegetation, macroinvertebrate, and am-
phibian data, and will form the basis for wet-
land biocriteria.
2. Identify and describe reference wetlands in
Ohio's four main ecoregions; Eastern Cornbelt
Plains, Erie/Ontario Drift and Lake Plain, Hu-
ron-Erie Lake Plain, and Western Allegheny
Plateau. These reference wetlands will be used
to develop biocriteria and also as "goals" for
wetland mitigation proj ects.
63
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3. Continue to assess whether the Ohio Rapid
Assessment Method correlates well with the
more in-depth measures of wetland quality, and
to test and refine breakpoints between the wet-
land categories.
4. Begin to assess the sensitivity of different meth-
ods in evaluating the relationship between wet-
land quality and the degree of disturbance
The State of Ohio has well-developed biological
criteria (or biocriteria) for streams, such as the In-
vertebrate Community Index (macroinvertebrates),
the Index ofBiological Integrity (fish), and the Modi-
fied Index of Well Being (fish) (Ohio EPA 1988a,b
and 1989). These indices are codified in Ohio
Administrative Code Chapter 3745-1. Until re-
cently, however, surface waters of the State that
are jurisdictional wetlands were only genetically
protected under Ohio's water quality standards.
On May 1,1998, the State of Ohio adopted wet-
land water quality standards and a wetland
antidegradation rule. These wetland quality stan-
dards developed narrative criteria for wetlands and
created the "wetland designated use." All jurisdic-
tional wetlands are assigned the "wetland designated
Ecoregions** of Ohio and Indiana and Neighboring States
so.
51.
53.
54.
55
nn 56.
El 57.
'in
m
I
I
D 61.
Notthem Lakes and Forests
IJorth Central Ha/dwood Forests
Southeastern Wisconsin Till Plains
Genital Can Bell Plains
Eastern Com Bell Plains
S. Michigan!N. Indiana 1111 Plains
Huron/Erie Lake Plains
Northern Appalachian Plateau and Uplands CH
Erie/Ontario Lake Hills and Plain
62. North Genital Appalachians
65. Southeastern Rains
66. Blue Ridge Mountains
67. Central Appalachian Ridges and Valleys
69. Cenlral Appalachians
70. Western Allegheny Plateau
71. Interior Plateau
72. Interior River Lw/tand
"Inset shows ecoregion boundaries Item USEPA I9S5 (Map M-l
FIGURE 17: ECOLOGICAL REGIONS OF OHIO AND INDIANA
(FROM U.S. EPA 1995).
64
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use." The State of Ohio did not attempt at this
time to identify multiple wetiandfimctions as wet-
land uses., because of the lack of data to develop
quantitative water quality criteria for wetlands.
However, the development of such biocriteria is the
ultimate goal and the primary thrust of this project.
The key part of Ohio's current regulatory pro-
gram for wetlands is found in the wetland
antidegradation rule. The wetland antidegradation
rule categorizes wetlands based on their functions,
sensitivity to disturbance, rarity, and irreplaceabil-
ity, and scales the strictness of avoidance, minimi-
zation, and mitigation to a wetland's category.
Three categories were established: Category 1
wetlands with minimal wetland function and/or in-
tegrity; Category 2 wetlands with moderate wet-
land function and/or integrity; and Category 3 wet-
lands with superior wetland function and/or integ-
rity. In order to implement the wetland standards
and antidegradation policy, wetlands must be as-
sessed on their relative quality. Ohio EPA has de-
veloped a draft Ohio Rapid Assessment Method.
The Ohio Rapid Assessment Method has proved
to be a fast, easy-to-use procedure for distinguish-
ing between wetlands of differing quality. How-
ever, it does not and was not intended to substitute
for direct, quantitative measures of wetland func-
tion (i.e., biocriteria).
Ohio began development of sampling methodolo-
gies and began sampling reference wetlands for
biocriteria development in 1996. To date, Ohio
has sampled 56 wetlands located primarily in the
Eastern Cornbelt Plains Ecoregion located in cen-
tral and western Ohio. These wetlands have in-
cluded depressional emergent, forested, and scrub-
shrub wetlands; floodplain wetlands; fens; kettle
lakes; and seep wetlands. The wetlands being stud-
ied span the range of condition from "impacted"
(i.e., those that have sustained a relatively high level
of disturbance) to "least-impaired" (i.e., the best
quality sites available).
On the basis of results to date (see Fennessy et
al. 1998a,b; Mack et al. unpublished data), Ohio's
research supports the use of vascular plants,
macroinvertebrates and/or amphibians as biologi-
cal metrics in wetlands, and also the continued use
and development of the Ohio Rapid Assessment
Method as a rapid assessment tool.
This work has been funded since 1996 by several
EPA Region 5 Wetland Program Development
Grants.
STUDY DESIGN
Fifty-seven wetlands were sampled during the
1996, 1997, 1998, and 1999 field seasons. The
first 2 years of data laid the groundwork for stan-
dardizing sampling methodologies, classifying wet-
lands, identifying potential attributes, and develop-
ing metrics using vascular plants, amphibians, and
macroinvertebrates.
In 1996, Ohio EPA monitored a series of riparian
forested across a gradient of disturbance (i.e., least
impacted to impaired) (Fennessy et al. 1998b).
Estimates of the relative level of disturbance were
made on a scale of 1 (most disturbed) to 10 (least
disturbed) based on visual evidence of disturbances,
review of aerial photographs of the wetland and the
surrounding area, and interviews with staff from the
Natural Resource Conservation Service and/or the
landowner. In 1996 and 1997, Ohio EPA moni-
tored 21 forested and emergent depressional wet-
lands. Relative disturbance was evaluated using a
tiered flow chart to assign a relative disturbance
score and also with the score from the Ohio Rapid
Assessment Method (Fennessy et al. 1998a, Fig-
ure 2.2).
Ohio EPAfound a good correlation between the
scores of the Ohio Rapid Assessment Method score
and level of disturbance a wetland site has experi-
enced. Higher ORAM scores correlate well with
65
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PLATE 7: IMPAIRED WETLAND.
PHOTO: MICKMICACCHION
PLATE 8: LEAST-DISTURBED WETLAND.
PHOTO: MICKMICACCHION
lower levels of disturbance based on our model, as
do lower ORAM scores with disturbed sites. In
1999, the ORAM score of the site was used as a
measure of the level of disturbance. So far, this
appears to be a highly effective "x-axis" disturbance
gradient for the development of IBIs for wetland
plants.
Reference wetlands are sites or data sets from
sites that typify a class of wetlands within a rela-
tively homogeneous physiographic region. Refer-
ence sites should include wetlands that have been
degraded or disturbed. Site selection in this study
is made using an ecoregional approach and to re-
flect a gradient of disturbance (i.e., least-impacted
to impaired).
Reference sites are selected such that relatively
similar proportions of low-, medium-, and high-dis-
turbance sites are sampled. To date, almost all the
wetlands studied by Ohio EPAhave been located
in the Eastern Cornbelt Plains (ECBP) ecoregion.
For the years 2000 and 2001 field seasons, Ohio
EPA will be studying reference sites in the Erie
Ontario Lake Plain (EOLP) ecoregion of northeast-
ern Ohio.
SAMPLING METHODS AND ANALYSIS:
MACROINVERTEBRATES AND
AMPHIBIANS
Below is a detailed description of the sampling
methodology and analysis process used by Ohio
EPAfor macroinvertebrates and amphibians. Also
included are the lessons learned for sampling these
assemblages.
Amphibian and macroinvertebrate taxa were se-
lected as potential indicators of wetland condition.
On the recommendation of many field profession-
als, experienced in amphibian monitoring, funnel
traps were used. The funnel traps also proved to
be extremely effective at sampling wetland
macroinvertebrate communities. Therefore, the
same sampling protocols are used for both amphib-
ians and macroinvertebrates in Ohio EPA's study.
Funnel trap
The funnel traps used in this proj ect have cylin-
ders constructed of aluminum window screen and
funnel ends made from fiberglass window screen.
The funnel traps are similar in design to commer-
cially available minnow traps. The cylinders are 18
inches long and 8 inches in diameter. The two fun-
nel ends are attached to the cylinders and begin 8
inches in diameter and taper inward 5 inches to a
P/4-inch opening. A string handle that runs from
end to end is attached to the two seams where the
cylinder and funnels ends j oin.
66
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To date, Ohio sampling data have come from de-
pressional wetland systems in the Eastern Corn Belt
Plains (ECBP) ecoregion. The Eastern Corn Belt
Plains ecoregion spans most of the western half of
Ohio and accounts for about 40% of Ohio's land
mass. Within the ECBP, depressional study wet-
lands that demonstrated a gradient of disturbance
levels, from least impacted to greatly disturbed, were
selected.
Selected wetlands are sampled for amphibians and
macroinvertebrates three times during the year
(early, middle, and late spring). An early spring
sample is collected in the period from late February
to late March. This early spring sample run allows
sampling of adult Ambystomatid salamanders and
early spring macroinvertebrates (e.g., fairy shrimp)
that are only present in wetlands for a limited time.
Adult salamanders enter wetlands to breed in early
spring following the first few warm, rainy nights of
late winter and early spring. The actual timing of
their arrival in Ohio is highly weather dependent and
varies greatly by year and location. A second
sample is collected during the month of April in or-
der to collect adult frogs entering wetlands to breed,
as well as amphibian larvae already present, and to
sample for macroinvertebrates. Mosquito larvae,
an important prey item for many predators, are
abundant in April in Ohio wetlands. The final sam-
pling is conducted between May 15 and June 15.
Salamander larvae and frog tadpoles are collected,
in addition to the resident macroinvertebrates. This
last sampling run occurs late enough in the breeding
cycle to allow collection of larvae from all breeding
amphibians. However, it is still early enough that
even in drought years temporary wetlands will not
have dried up.
Generally, 10 funnel traps are installed at each
wetland. Prior to installing the first funnel trap, the
perimeter of the area where standing water is
present is measured using a hip chain or by pacing.
The total perimeter length is then divided by 10,
and 10 funnel traps are installed uniformly around
the perimeter of the wetland at intervals of 10% of
the total perimeter distance. Each funnel trap loca-
tion is permanently marked with flagging for use
throughout the sampling season. The funnel traps
are installed on the bottom at a location deep enough
to submerge the trap. The traps are left in the wet-
land 24 hours to ensure unbiased sampling for ani-
mals with diurnal and nocturnal activity patterns. The
design of the traps allows them to collect any am-
phibians and macroinvertebrates that swim or crawl
into the funnel openings.
Upon retrieval, the traps are emptied by inverting
one of the funnels and dumping and shaking the
contents into a white sorting pan. Organisms that
can be readily identified in the field are counted and
recorded in the field notebook and released. The
remaining organisms are transferred to a 1 -L plas-
tic bottle and preserved with 70% ethanol. The
contents of each trap are kept in separate, clearly
marked bottles for individual analysis in the labora-
tory. If large numbers of amphibians are kept for
identification in the lab, the samples are transferred
to formalin for long-term storage. All organisms
collected are identified in the lab using appropriate
keys and the results are recorded.
Dipnet
Qualitative collections are made concurrently with
funnel trapping at each wetland once during each of
the three sampling periods. Qualitative sampling
involves the collection of amphibians and
macroinvertebrates from all available natural wet-
land habitat features. This is achieved by using tri-
angular ring frame 30-mesh dipnets and manual
picking of substrates with field forceps. The goal is
to compile a comprehensive species/taxa list of
macroinvertebrates and amphibians at the site. A
minimum of 30 minutes is spent collecting the quali-
tative sample. Sampling continues until the field crew
determines that further sampling effort will not pro-
duce new taxa. At least one specimen of all taxa
collected during the qualitative sampling is preserved
in a j ar of ethanol for positive identification in the
laboratory.
67
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Hester-Dendy artificial substrate sampler
FiveHester-Dendy (HD) artificial substrate sam-
plers were tied to the top of a concrete block and
placed in wetlands where they remained submerged
for 6 weeks. The samplers were collected and pre-
served in formalin. All the macroinvertebrates colo-
nizing the samplers were counted and identified.
ANALYTICAL METHODS:
MACROINVERTEBRATES AND
AMPHIBIANS
Macroinvertebrates were identified to genus or
species. Amphibians were identified to species ex-
cept for some small salamander larvae identified to
genus. Each funnel trap collection was analyzed
individually so that location-specific information was
not lost by pooling all samples from a site.
The number of individuals collected in the traps
was divided by the number of hours trapped to give
a relative abundance consisting of number of indi-
viduals per trap hour. Results from the different
wetland study sites were examined for faunal dif-
ferences in distribution and abundance. Analysis of
the data for potential biological indicators of human
disturbance is under way.
LESSONS LEARNED FOR
MACROINVERTEBRATES AND
AMPHIBIANS
Funnel traps consistently collected an average
of 10 more macroinvertebrate taxa than quali-
tative sampling using dipnets. Funnel traps were
much more effective in sampling amphibians and
fish than sampling with dipnets.
Qualitative sampling collected somewhat more
Mollusca and Chironomidae taxa than did fun-
nel traps.
Funnel traps collected more leech taxa, Hemi-
pterataxa, Col eoptera taxa, Odonatataxa, and
Crustacea taxa than did qualitative sampling.
Hester-Dendy artificial substrate samplers were
ineffective for sampling most wetland
macroinvertebrates, except oligochaetes,
Chironomidae, and Mollusca.
A 24-hour sampling period for funnel traps is
preferred as it allows for the collection of noc-
turnal species that are infrequently collected by
daytime sampling methods.
A single wetland often has several vegetation
classes. Even if only three main classes are identi-
fied (forested, scrub-shrub, and emergent), the
wetlands included for study can exhibit multiple com-
binations. For example, of the 20 wetlands studied
inFennessy etal. (1998a), 6 combinations of veg-
etation classes were found: emergent, emergent/
scrub-shrub, forested, forested/emergent, forested/
emergent/scrub-shrub, and forested/scrub-shrub.
Thus, a sampling method should be flexible enough
to account for horizontal and vertical variation in
vegetation.
After testing a transect-quadrat method, Ohio EPA
has adapted the method used by the North Caro-
lina Vegetation Survey as its standard vegetation
sampling method (Peet et al. 1998). This is a flex-
ible, multipurpose sampling method that can be used
to sample such diverse communities as grass- and
forb-dominated savannahs, dense shrub thickets,
forest, and sparsely vegetated rock outcrops, and
has been used at more than 3,000 sites over 10
years as part of the North Carolina Vegetation Sur-
vey. This method is appropriate for most types of
vegetation, flexible in intensity and time commitment,
compatible with other data types from other meth-
ods, and provides information on species compo-
sition across spatial scales. It also addresses the
problem that processes affecting vegetation com-
position differ as spatial scales increase or decrease
and that vegetation typically exhibits strong
autocorrelation (Peet etal. 1998). Peet etal. (1998)
state, "Our solution to the problems of scale and
spatial auto-correlation is to adopt a modular ap-
proach to plot layout, wherein all measurements are
68
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made in plots comprised of one or more lOxlO-m
quadrats or "modules" (100 m2 = 1 are = 0.01 hect-
are). The module size and shape were chosen to
provide a convenient building block for larger pots,
and because a body of data already exists for plots
of some multiple of this size. The square shape is
efficient to lay out, ensures the observation is typi-
cal for species interactions at that scale of observa-
tion, and avoids biases built into methods with dis-
tributed quadrats or high perimeter-to-area ratios"
(Peet et al. 1998, p. 264). Basically, the method
employs a set of 10 modules in a 20x50-m layout.
Within the site to be surveyed, these 20x50-m grids
are located such that the long axis of the plot is
oriented to minimize the environmental heterogene-
ity within the plot.
Once the plot is laid out, all species within the plot
are identified, an aggregate wood stem count is made,
and cover is estimated at the 0.1 -hectare scale. In
addition, four lOx 10-m modules are intensively
sampled in a series of nested quadrats. Within these
"intensive" modules, species cover class values and
woody stem tallies are recorded for each module
separately and for each nested quadrat separately.
In effect then, the method proposed by Peet et al.
incorporates use of releves found in the Braun-
Blanquet methodology inasmuch as the length,
width, orientation, and location of the modules are
qualitatively selected by the investigator on the ba-
sis of site characteristics; however, within the mod-
ules, standard quantitative floristic and forestry in-
formation is recorded, such as density, basal area,
cover, etc.
Once the location of the plot or plots has been
selected, the primary purpose of the vegetation sur-
vey is to obtain a comprehensive list of all vascular
plant species growing at a particular wetland at the
time of sampling and to characterize the relative
dominance of these species at several levels of scale
(basically herbaceous, shrub, small tree, and large
tree scales, or at 1 m2,10 m2, 100 m2, and 0.1 ha
(1,000m2 or 1 are).
All vascular species within the modules are iden-
tified to species. Immature plants or plants missing
structures (e.g., fruiting bodies, etc.) that cannot be
identified to species are identified to genus or fam-
ily or noted as unknown. Within the intensively
sampled modules, percent cover is recorded for
each species within modules and nested quadrats.
Cover classes suggested by Peet etal. (1998) are
used as a faster and more repeatable method for
assigning cover values: Class 1 (solitary/few), Class
2 (0 to 1%), Class 3 (1% to 2%), Class 4 (2%-
5%), Class 5 (5%-10%), Class 6 (10%-25%),
Class 7 (25%-50%), Class 8 (50%-75%), Class 9
(75%-95%), Class 10 (95%-99%). The midpoints
of the cover classes are used to calculate percent
cover, relative cover, etc.
Woody stem data (trees, shrubs, and woody li-
anas reaching breast height or 1.4 m) are collected
as counts of individuals in diameter classes. Peet
etal. (1998) suggest the following diameter classes
(in cm): 0-1, 1-2.5, 2.5-5, 5-10, 10-15, 15-20,
20-25, 25-30, 30-35, and 35-40, with stems
greater than 40 cm counted individually and mea-
sured to the nearest centimeter. Multiple stems aris-
ing from a common root system are recorded sepa-
rately if they branch below 0.5m from the ground.
Peet et al. (1998) recommend that the area sur-
veyed by stem count be adjusted based on condi-
tions at the site, e.g., reduced to 20% of the mod-
ules for dense shrubland or increased by 200% for
oak savannahs. This is easily implemented by re-
ducing the width of the modules for woody species
only.
An important part of vegetation surveys is collec-
tion, preparation, and depositing of voucher speci-
mens in maj or herbariums in order to document a
permanent record of that plant at that location. Al-
though staff resources make collecting vouchers of
every vascular plant infeasible, a voucher specimen
of at least 10% of the vascular plant species at any
given site is collected; however, in every instance in
which the identity of any species cannot be con-
firmed in the field, or where field personnel disagree
69
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as to the identity of a species, a voucher specimen
is collected for identification in the office. In par-
ticular, difficult genuses and families (e.g.,
Cyperaceae, Poaceae, Ranunculaceae, Viola, As-
ter, Potamogeton), as well as endangered, threat-
ened, rare, or otherwise unusual plants, are almost
always collected for confirmation.
Finally, data on standing biomass for emergent
wetlands are collected. These data can be used in
several ways. Biomass production in emergent
wetlands dominated by herbaceous vegetation is
estimated by harvesting 900-cm2 quadrats in each
wetland. The quadrats are located within the inten-
sive modules of each plot. The plants within each
quadrat are cut at the soil surface and placed into
paper bags. In the lab, plants are oven dried at
105°C for at least 24 hours and then weighed.
LESSONS LEARNED FOR
VASCULAR PLANTS
Floristic Quality Assessment Indexes
Ohio EPA has found that the FQAI score and
subscores of the FQAI, e.g., percent coverage of
plants with coefficients of conservatism of 0, 1,
or 2, are very successful attributes and metrics
for detecting disturbance in wetlands. See the fol-
lowing references:
Andreas B, LichvarR. 1995. AFloristic Quality
Assessment System for Northern Ohio. Wetlands
Research Program Technical Report WRP-DE-8.
U. S. Army Corps of Engineers, Waterways Experi-
ment Station.
Herman KD, Masters LA, Penskar MR, Reznicek
AA,WilhelmGS,BrodowiczWW. 1996. Floris-
tic Quality Assessment With Wetland Categories
and Computer Application Programs for the State
of Michigan. Michigan Department of Natural Re-
sources, Wildlife Division, Natural Heritage Pro-
gram.
Wilhelm GS, Ladd D. 1988. Natural area as-
sessment in the Chicago region. Transactions 53rd
North American Wildlife and Natural Resources
Conference, pp. 361- 375.
Wilhelm GS, Masters LA. 1995. Floristic Qual-
ity Assessment in the Chicago Region and Applica-
tion Computer Programs. Lisle, IL: Morton Arbo-
retum.
Semiquantitative disturbance/integrity scales
Ohio EPA has had good success in developing a
semiquantitative disturbance/biological integrity scale
called the Ohio Rapid Assessment Method for
Wetlands v. 5.0. Until more quantitative variables
such as percent impervious surface are found, this
type of tool is a good candidate for the problematic
x-axis in wetland biocriteria development. See also
"Plants and Aquatic Invertebrates as Indicators of
Wetland Biological Integrity in Waquoit Bay Wa-
tershed, Cape Cod," Carlisle BK, Hicks AL, Smith
JP, Garcia SR, Largay BG Environ Cape Cod
1999; 2(2):30-60, where a similar system was used
to rank levels of disturbance.
Classification
Classification is definitely an iterative process.
Investigators should consider a hydrogeomorphic
(HGM) classification scheme if one has been de-
veloped for their region of interest, at least as a start-
ing point. However, the experience in Ohio sug-
gests that grosser classes based on dominant veg-
etation (emergent, scrub-shrub, forested, etc.) may
work also. A goal of a cost-effective biocriteria
program is to have the fewest classes that provide
the most cost-effective feedback. With vegetation,
data from Ohio are suggesting that somewhat di-
verse wetland types may be "clumpable," because
even though their flora are different at the species
level, the quality/responsiveness of their unique flora
to human disturbance is equivalent. This is also a
concern in States with high degrees of wetland loss
where too few wetlands of a particular HGM class
remain to analyze as a separate class.
7O
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Field and lab methods
After experimenting with both transect/quadrat and
releve-style plot methods, Ohio has adopted a plot-
based method that allows for a qualitative stratifi-
cation of wetland by dominant vegetation commu-
nities. This method is flexible and adaptable to
unique site conditions, provides dominance data for
all species in all strata, provides data that are
intercomparable with other common methods, is
relatively easy to learn, and is relatively fast and
cost-effective (up to two to three plots can be com-
pleted in a day).
Whatever sampling method is adopted, it is es-
sential that dominance and density information
(cover, basal area of trees, stems per unit area, rela-
tive cover, relative density, importance values, etc.)
be collected. Many of the most successful attributes
Ohio has found in developing a vegetation IB I are
based on cover data of the herb and shrub layers
and density data of the shrub and tree layers.
Definitely consider using cover classes in general
and a class scheme that works on a doubling prin-
ciple to aid in consistent inter-investigator usage (see
Peetetal. 1998). Then use the midpoints of the
class for your analysis. This seems to help with
consistent usage and smoothing out the roughness
in cover data.
Finally, it is recommended that initially the
sampling method should "overstratify" in both the
vertical and horizontal dimensions until it can be de-
termined which strata and communities are respond-
ing best to human disturbance. Ohio has found that
the herb and shrub (subcanopy layers) seem to
respond the best, although some intermediate tree
size classes (e.g. 10- to 25-cm dbh) also appear
responsive.
Overstratifying horizontally may also make sense
at the reference development stage; however, ulti-
mately the decision whether to split or clump com-
munities depends on whether this is necessary to
detect the disturbances. "Homogenizing" commu-
nity types by placing a releve or transect across
them (e.g., aquatic bed to emergent to shrub zone)
can be appropriate if splitting does not matter to
detect the disturbance. The caveat, of course, is
that you cannot separate the data set later if you
detect something of interest in one of the clumped
communities.
Vouchers and QAQC
On the basis of Ohio's experience, voucher as
much as you can for later confirmation in the lab
and deposit vouchers in local and regional
herbariums. Definitely collect all Cyperaceae,
Poaceae, and Juncaceae, and also consider col-
lecting shrubs genuses and families (Salix, Vibur-
num, Vaccinium, Rosa, Alder, etc.), Polygonum
spp., Aster spp., Viola spp., and Cryptograms.
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OREGON: SIMULTANEOUS DEVELOPMENT,
CALIBRATION, AND TESTING OF HYDROGEOMORPHIC-
BASED (HGM) ASSESSMENT PROCEDURES AND
BIOLOGICAL ASSESSMENT PROCEDURES
Contact
PaulAdamus,Ph.D
Organization Adamus Resource Assessment,
Inc.
6028 NW Burgundy Drive
Corvallis, OR 97330
Phone (541) 745-7092
E-mail adamus7@attbi.com
Contact
Dana Field
Organization Oregon Division of State Lands
775 Summer St. NE
Salem, OR 97310
Phone (541)378-3805, ext. 238
E-mail Dana.Fi el d@dsl. state, or.us
PURPOSE
Collect and analyze field data on wetland and
riparian plant communities and hydrogeo-
morphic (HGM) features to define reference
standard conditions and performance criteria
for four subclasses of wetlands in Oregon, two
in the Willamette Valley in western Oregon and
two on the Oregon coast
Use that information to develop quantitative but
rapid, HGM-based assessment models and
procedures for those subclasses, as well as to
identify new wetland plant community metrics
that have demonstrated relationships with land-
scape condition and with internal alterations of
wetlands in this region
Compare results of using HGM models for func-
tions with metrics that describe Holistic charac-
teristics, to verify the results on independent data
sets, and demonstrate implementation of the
methods for assessing progress of restored
wetlands and for helping recommend compen-
sation ratios in a wetland mitigation bank con-
text
WETLAND TYPE
Slope, flats, and estuarine fringe wetlands
ASSEMBLAGES
Vascular plants
STATUS
Completed reports for the two Willamette Valley
subclasses, and currently initiating development
of the estuarine fringe project. Information on
purchasing the reports can be found at:
http://statelands.dsl.state.or.us/hgm_
guidebook.htm.
PROJECT DESCRIPTION
These proj ects are being done cooperatively with
EPA Region 10 and the Oregon Division of State
Lands, which in 1997 identified a need, in the con-
text of Section 404 and 401 responsibilities, for a
more quantitative assessment method tailored spe-
cifically to these wetland subclasses. The proce-
72
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dures have been designed to be applicable to
nonjurisdictional riparian sites as well as to wet-
lands. The proj ect began in 1998 with formation of
an interagency oversight committee, intensive re-
view of regional literature, and reconnaissance and
preliminary selection of 124 reference sites through-
out the Willamette Valley ecoregion. In 1999, the
list of sites was narrowed, workshops of local ex-
perts were held to develop function models and field
procedures, 38+ volunteers were recruited and
trained to assist with field work, and 2 assessment
teams ("A-teams") of scientists with help from the
volunteers collected data from the selected sites. A
three-volume final report was published in early
2001 (see above). The report for the estuarine
fringe wetlands is scheduled for completion in 2004.
Under administrative rules recently reissued by the
Oregon Divi sion of State Lands, applicants for most
State-authorized wetland removal-fill permits are
now required to use the regionalized HGM classifi-
cation that was developed by this proj ect, and are
encouraged to use the reference-based function
assessment procedures when available for a par-
ticular subclass and region of the State.
STUDY DESIGN
For the Willamette Valley project, plant and
hydrogeomorphic data were collected from 109
wetland and riparian sites (54 riverine impounding
subclass, 55 slope/flats subclass). The "riverine
impounding" sites include all wetland and riparian
areas within a 2-year river floodplain, e.g., sloughs,
oxbows, cut-off channels, beaver impoundments,
stream-fed ponds with water control structures.
"Slope/Flats" sites include most ash swales, wet
prairies, springs, and foothill seeps. From visual
inspection alone, most sites in the "slope" HGM
class could not be reliably separated from most sites
in the "flats" HGM class, due to the flat topography
and varied geology of the region, so the two were
combined into a "slope/flats" subclass.
Within the subclasses, assessment sites were se-
lected nonrandomly, in a manner intended to span
gradients of human disturbance, size, and plant com-
munity succession. Restored/created sites of vari-
ous ages were also included. Nearly all sites were
on public lands, and ranged in size from 0.1 to 233
acres. Most sites were nominated by local wetland
experts. From the collected data, five least-dis-
turbed riverine sites were selected as reference stan-
dards, as were six least-disturbed slope/flat sites.
Use of cluster analysis and other statistical analysis
methods verified that sites belonging to the two main
subclasses were significantly different based on land-
scape position and mapped soil characteristics im-
portant to ecosystem function.
For defining the disturbance gradient (the "x axis"),
information specific to each candidate site was col-
lected during the reconnaissance phase. This infor-
mation pertained to past management practices,
surrounding land use, and recent physical alterations.
Physical alterations were noted visually during re-
connaissance visits. At each site each type of alter-
ation was categorized as (a) absent, (b) physically
affects <10% of site, (c) affects >10% of site but
not all, or (d) affects entire site. The categories that
were used for alterations were:
Flow-impounding - berms, dikes, dams
Flow-impounding - excavations, pits
Flow-redirecting
Water subsidy (e.g., stormwater pipes)
Drainage-inducing (ditches, tile)
Soil compacting (e.g., fill, machinery, cows)
Soil mixing (e.g., plowing)
Soil grading (e.g., flattening)
Vegetation removal (e.g., extreme grazing, log-
ging)
SAMPLING METHODS: PLANTS
At each site, the A-teams identified plants in each
of potentially three hydrologic zones: permanent
water zone, seasonally inundated zone, saturated-
73
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only zone. For woody plants, the team walked the
entire site and made an overall estimate of relative
percent of the area of each zone occupied by each
shrub species (understory and open) and tree spe-
cies. For herbaceous plants, the team assessed
relative cover of each species in 1x1-m quadrats.
No more than nine quadrats were used at any site
(and fewer at smaller sites and sites with fewer hy-
drologic zones). Larger numbers of quadrats were
not used because of time and resource constraints.
Within zones, quadrats were located so as to maxi-
mize the cumulative number of species found. This
less formal approach was used because our goal
was to develop metrics based mainly on commu-
nity composition rather than quantitative measures
of abundance or cover. Two teams assessed all
109 sites in about 50 workdays.
ANALYTICAL METHODS: PLANTS
From the collected data, the following plant metrics
were compiled for each site:
Number of native herb species, relative to the
intensity of sampling (# of plots at each site)
(based on analysis using species-area curves
and regression)
Percent of herb species that are native
Percent of the herb species that are "remnant-
dependent" (i.e., reputedly most characteristic
of unaltered sites)
Percent of the dominant herb species in any plot
that are natives
Percent of the dominant herb species in any plot
that are remnant-dependent
Percent of the true wetland species (facultative
or wetter) that are natives
Percent of shrub species that are natives (when
shrubs present)
Percent of native shrub species that are at least
moderately dominant
Percent cover of non-native shrubs
Preliminary analysis suggests many of the above
metrics had a statistically significant association with
categorical observations of partial physical degra-
dation of sites (see Study Design, above) and/or
with amount and proximity of surrounding land cover
categories (agriculture, urban, natural) that were
assessed visually during fieldwork as well as by a
GIS analysis of existing digital imagery. Analyses
of the HGM and plant data sets, each containing
4,000+ records, were performed on a PC using
Excel, PC-ORD, andNCSS.
LESSONS LEARNED
Random or systematic sampling, whether within
a region or within a site, is not always appro-
priate for use in identifying good biological in-
dicators or developing rapid models for assess-
ing wetland condition and function.
Systematic, repeatable, rapid procedures can
be developed for assessing some of the distur-
bance gradients. This is a necessary precursor
to selecting reference sites that will yield the most
useful data.
The biological metrics investigated or used
should be appropriate to the study design and
measurement protocols.
Data suitable for identifying biological indica-
tors of wetland condition sometimes can be col-
lected simultaneously with data collected for
calibration of HGM models. This does not nec-
essarily require a great deal of additional train-
ing or field time.
Shared field experiences are a good forum for
sharing wetland knowledge among agencies, and
among agencies and consultants and citizens.
Shared field experiences lead to participants feel-
ing more vested in the process of developing mod-
els and multimetric indexes. This informal "buy-in"
can lead to greater willingness of participants to use
the methods that ultimately result.
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PENNSYLVANIA: ASSESSING WETLAND CONDITION AT
A WATERSHED SCALE USING HYDROGEOMORPHIC
MODELS AND MEASURES OF BIOLOGICAL INTEGRITY
Contact
Rob Brooks
Contact
Denice Heller-Wardrop
Organization Penn State University
Penn State Cooperative Wetlands
Center
301 Forest Resources
Laboratory
University Park, PA 16802
Phone (814)863-1596
E-mail rpb2@psu.edu
Website http://www.wetlands.cas.psu.edu/
Organization Penn State University
Penn State Cooperative Wetlands
Center
301 Forest Resources
Laboratory
University Park, PA 16802
Phone (814)863-1005
E-mail dhwl 10@psu.edu
Website http://www.wetlands.cas.psu.edu/
PURPOSE
Develop biological and functional assessment
methodologies for many wetland types in Penn-
sylvania
WETLAND TYPE
Variety of wetland types
ASSEMBLAGES
Amphibians
Birds
Macroinvertebrates
Vascular plants
PROJECT DESCRIPTION
The Penn State Cooperative Wetlands Center
(CWC) has numerous ongoing proj ects that involve
bioassessment methodology for assessing the con-
dition of wetlands. These proj ects include:
Bird community index of biological integrity for
the Mid-Atlantic Flighlands
Verified suitability index for the Louisiana wa-
terthrush
Amphibian indicator landscape study
Macroinvertebrates indicator pilot study
Otter and beaver interactions in the Delaware
Water Gap
Using bioindicators to develop a calibrated in-
dex of regional ecological integrity for forested
headwater systems
In addition to these proj ects, CWC is conducting
a 2-year pilot study to assess the ecological condi-
tion of wetlands in the Juniata River watershed of
central Pennsylvania. The Juniata River is a major
tributary of the Susquehanna River and lies within
the headwaters region of the Chesapeake Bay eco-
system. The objective of the Juniata Wetland Moni-
toring Project is to define the ecological health of
wetland resources, thereby providing a scientific
context for resource managers to plan future pro-
tection and restoration activities.
75
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CWC is using HGM functional assessment mod-
els that incorporate a floristic quality assessment
index (FQAI) similar to the one used by Ohio EPA
(see Ohio EPA's case study). They use a three-
step process. The first step is to develop and cali-
brate HGM functional assessment models that are
sensitive to environmental disturbance. The sec-
ond step is to randomly sample wetlands within the
watershed that represent a range of HGM classes
and disturbance levels. The final step is to apply the
HGM models to the data collected in the field to
determine the condition or health of wetlands within
the Juniata River watershed.
Preliminary HGM functional assessment models
have been developed by the CWC. These models
are currently being calibrated using a set of 102
reference wetlands that span a variety of HGM sub-
classes (mainstem floodplain, headwater floodplain,
riparian depression, slope, etc.) and disturbance
levels (severe, moderate, or pristine). Disturbance
levels are assigned based on the surrounding land-
scape, the type of buffer present, and potential stres-
sors identified at the site. It is essential to charac-
terize wetlands across a disturbance gradient to
determine not only the level of function a given wet-
land type may achieve, but also the level of func-
tioning that is attainable in an impacted landscape.
The second step of the process, data collection,
is being undertaken by the CWC and trained in-
terns from Perm State University, the University of
Pittsburgh at Johnstown, and the Department of
Environmental Protection. Two types of data are
collected for each wetland sampled: landscape-
level data and site-level data. Landscape-level data
are obtained at the CWC by characterizing land
uses within a 1-km radius circle surrounding the
wetland using aerial photographs. This gives an in-
dication of the type and magnitude of potential wet-
land stressors. For example, activities such as land
clearing in the surrounding watershed may stress
wetland systems by increasing sediment loads.
In addition to land uses, the buffer type and width
is also determined for each site. It is important to
look at buffers when studying disturbance because
buffers can ameliorate or magnify the affects of nega-
tive landscape activities. A wetland surrounded by
a forested buffer may not be as stressed by activi-
ties in the landscape as one surrounded by an ur-
ban or agricultural buffer.
Site-specific data are collected at each wetland
site in the field. Site-level responses of wetlands to
stressors are many and may involve both abiotic
and biotic indicators. At each wetland sampled, data
are gathered on microtopography, soils, plants, and
animals. Although not all plant species are highly
sensitive to disturbance, the immobility of the plant
community, its amenity to remote sensing tech-
niques, and easily recognized signs of stress make
it a good indicator of wetland condition. Previous
studies by the CWC have investigated the potential
utility of plant community measures as indicators of
wetland health. Plant community measures that may
prove to be good indicators of disturbance include
the FQAI score, the number or dominance of in-
troduced or aggressive native species, and the num-
ber of different species of Carex present. The CWC
method contains a comprehensive plant sampling
methodology that has been used in a variety of
project types. Three sizes of plots are used to
record various measures of the plant community
including percent cover and richness of herbaceous
species, shrub volume, and tree dbh.
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VERMONT: CLASSIFICATION, BIOLOGICAL
CHARACTERIZATION, AND BIOMETRIC DEVELOPMENT
FOR NORTHERN WHITE CEDAR SWAMPS
AND VERNAL POOLS
Contact
DougBumham
Organization Vermont Department of Environmental Conservation
WQD
103 S. Main St.-ION
Waterbury, VT 05676
Phone (802) 241-3784 or 244-4520
E-mail DOUGB@dec.anr.state.vt.us
PURPOSE
The Vermont Wetlands Bioassessment Proj ect is
a collaboration between the Vermont Department
of Environmental Conservation and the Vermont
Nongame and Natural Heritage Program, with a 3 -
year time frame. The primary obj ectives of the first
phase of the Vermont Wetlands Bioassessment Pro-
gram are:
Gather chemical, physical, and biological data
from seasonal pools that will facilitate an eco-
logically based classification of minimally dis-
turbed (reference) seasonal pools in Vermont
Use both previously and newly collected Non-
game and Natural Heritage Program data to
try to identify specific biological attributes that
can serve as indicators of ecological integrity in
northern white cedar swamps.
WETLAND TYPE
Vernal pools
White cedar swamps
ASSEMBLAGES
Algae
Amphibians
Birds
Macroinvertebrates
Vascular plants
STATUS
Analyzing data and writing final report. A final
report is expected in the near future.
PROJECT DESCRIPTION
The Vermont Wetlands Bioassessment proj ect cur-
rently focuses on two types of wetlands: (1) vernal
(seasonal/ephemeral) pools and (2) Atlantic north-
ern white cedar swamps.
Vernal pools
We sampled a total of 28 vernal pools for breed-
ing amphibians, macroinvertebrates, algae (diatoms),
soils, vegetation, and water chemistry in April, May,
and June of 1999 and 2000. A variety of other
physical and riparian observations were also made.
77
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Pools were distributed throughout Vermont across
seven biophysical regions. Arange of condition,
from "reference" or least disturbed, to highly dis-
turbed, was represented in the selection of pools.
Upon completion of the field phase of the project
and assessment of relative hydroperiod, it is likely
that 5 of the 28 pools may be less "seasonal" than
originally thought. Data are being analyzed for bio-
logical signals that may identify pools with more-
or-less permanent standing water. Each pool was
visited twice, the two visits approximately 4 to 6
weeks apart, for aquatic macroinvertebrate and
water chemistry sampling. Algae samples were col-
lected on the second pool visit. Amphibian surveys
either preceded or coincided with the first
macroinvertebrate sampling visit. Five of the 28
pools were dry on the second sampling visit.
Aquatic macroinvertebrates. Macroinverte-
brates were sampled using three different methods:
funnel traps to sample the actively swimming inver-
tebrates (i.e., beetles, bugs, mosquitoes, crusta-
ceans), a D-net to sample benthic invertebrates in
the leaf litter and muck (i.e., snails, bivalves, chi-
ronomids, oligochaetes, caddisflies), and a qualita-
tive search for any taxa we might have missed with
the previous two methods. Funnel traps were made
of window screen and designed to function like min-
now traps. The traps were placed approximately
10m apart and were left in place for approximately
24 hours. When the traps were emptied, any am-
phibians were returned to the pool and the
macroinvertebrates were collected and preserved.
The contents of each trap were stored separately.
The D-net scoop and qualitative samples were pre-
served in the field, and later picked and sorted into
taxonomic orders according to standard protocol.
Except for zooplankton, all taxa were identified to
the lowest practicable level. Scoop sampling along
transects often created more of a disturbance in the
pool than was comfortable for those conducting the
sampling. The presence of egg masses and/or large
concentrations of larval amphibians greatly restricted
the efficiency of the scoop sampling at many pools.
Trap sampling was less disruptive, but care had to
be taken to not completely submerge the traps when
adult amphibians were present so that entrapped
adults would have air space to breathe. High con-
centrations of mosquito larvae tended to affect the
efficiency of the traps. Qualitative sampling was
less disruptive (although excessive wading in the
pools often created an uncomfortable level of dis-
turbance) but provided an inconsistent sampling ef-
fort. The maj ority (70%) of macroinvertebrate taxa
collected were represented in trap samples. Two
orders represented a large proportion of all taxa
collected: Diptera (35%) and Coleoptera (23%).
Water. Water temperature, pH, and apparent
color were recorded in the field. The remainder of
the water sample was preserved and later analyzed
for alkalinity, conductivity, anions, cations, and alu-
minum, and again for pH. Field pH values ranged
from a minimum of 4.41 to a maximum of 7.75,
alkalinity values ranged from 0 mg/L to 173.0 mg/
L, conductivity readings ranged from 14.9 umhos/
cm to 335 umhos/cm, and apparent color ranged
from 4.5 to 489 over the sampling season. Appar-
ent color, alkalinity, and cation concentrations show
a general increasing trend within each site over the
sampling season, whereas pH and aluminum and
anion concentrations exhibited no consistenttrend.
Algae. Algae were collected and analyzed for
diatoms from 18 pools sampled in 1999. Algae sam-
pling primarily targeted diatoms; however, filamen-
tous algae were collected when present. We at-
tempted to collect both benthic samples (scraping
algae from leaves, sticks, and rocks) and plank-
tonic diatom samples from each pool. Unfortu-
nately, five of the pools were already dry in May,
so we were only able to collect planktonic samples
in the remaining 13 pools. Benthic samples were
collected from 17 of the 18 pools. We froze all
samples upon return to the lab, and samples were
sent to Dr. Jan Stephenson for identification.
Amphibians. We observed amphibians initially
during the amphibian survey, and continued to ob-
serve and collect specimens during both rounds of
78
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macroinvertebrate sampling. At the beginning of
the field season, we visually surveyed each pool for
egg masses and spermatophores, identified each egg
mass, recorded an approximate number of eggs per
mass, counted and identified breeding adults, and
described physical parameters of the habitat. The
timing of this first visit did not necessarily coincide
with amphibian emergence; however, the funnel traps
used during invertebrate sampling effectively caught
active amphibians. Breeding adults were typically
captured during the first round of sampling, whereas
tadpoles and larvae were present in the traps on
the second visit. All amphibians caught by the fun-
nel traps, including adults, larvae, and egg masses,
were identified in the field, counted, and returned
to the pool. We continued to survey egg masses
and record physical parameters throughout the field
season. All 28 pools sampled showed signs of use
by breeding amphibians. Amphibian species com-
monly observed included wood frogs (27/28 sites),
yellow-spotted salamanders (27/28 sites), Jefferson
(hybrid) salamanders (7/28 sites), red-spotted newts
(18/28 sites), and green frogs 15/28 sites).
Ongoing and upcoming work. Data are cur-
rently being evaluated using a variety of procedures
in order to identify and describe efficient and least
disruptive sampling methods, and evaluate biologi-
cal, chemical, and physical indicators of natural vari-
ability and disturbance. Methods of data analysis
include two-way indicator species analysis
(TWINSPAN) and detrended correspondence
analysis (DCA) as well as standard descriptive and
comparative statistical methods. Adetailed site re-
port is being developed for each vernal pool site.
Northern white cedar swamps
Seven northern white cedar swamps were sur-
veyed in June for breeding birds and vegetation.
Two of these cedar swamps were considered to be
of reference quality, whereas the other five had some
degree of impairment associated with them. Dis-
turbances at the impaired sites included logging,
roads, and storm water and agricultural discharge.
We visited three of the sites early in summer to
assess the feasibility of sampling aquatic
macroinvertebrates for bioassessment and monitor-
ing purposes.
Breeding birds/vegetation. Vegetation and bio-
physical data were collected at each site and spe-
cies lists are being constructed. Listening stations
were established and a bird census was taken twice
during the breeding season. The data collected from
the vegetative, biophysical, and bird sampling will
be compiled and compared to the existing data from
the statewide inventory of northern white cedar
swamps.
Aquatic macroinvertebrates. Sampling for
aquatic macroinvertebrates had been scheduled for
May, but because of a busy vernal pool sampling
season, we were unable to visit the cedar swamps
until June. Unfortunately, a very dry spring may
have caused the cedar swamps to be drier than usual
this year. We visited two impaired sites and one
reference site during the month of June. We were
unable to find any standing or flowing water at ei-
ther the reference site or at one of the impaired sites,
so we could not effectively sample. However, we
found evidence that suggested the swamps had con-
tained water earlier in the season. The second im-
paired site contained many braided, slow-flowing
channels and some standing water. We qualitatively
sampled three of the channels and a small hollow at
the base of the boulder. The samples were pre-
served in the field, picked, and sorted, and will be
identified according to standard protocol. It is pos-
sible that aquatic microhabitats in cedar swamps
are not available consistently enough to sample for
aquatic macroinvertebrates; however, our limited
sampling efforts did not conclusively elucidate the
feasibility of using aquatic macroinvertebrates as
biological indicators in cedar swamps.
Ongoing and upcoming work. Using the data
we have collected, we will work to identify attributes
associated with vegetation and bird assemblages
that can serve as indicators of ecological integrity
and anthropogenic disturbance.
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WASHINGTON: KING COUNTY WETLAND-BREEDING
AMPHIBIAN MONITORING PROGRAM
Contact Klaus O. Richter
Organization King County Department of Natural
Resources & Parks
Water and Land Resources Division
201 S Jackson Street, Suite 600
Seattle, WA 98104-3855
Phone (206) 205-5622
E-mail klaus.richter@metrokc.gov
Organization Viewpoint Services
11040104th Ave.NE
Kirkland,WA 98033-4423
Phone 425-822-4016
E-mail korichter@msn.com
PURPOSE
Provide yearly data regarding wetland condition
and the health of breeding amphibians for environ-
mental review, to identify threats to wetlands and
amphibians, to evaluate current wetland protection
measures, and to improve local-land use decisions.
Specifically, to stem amphibian extinctions in urban
landscapes by first identifying cause-and-effect re-
lationships among amphibian distribution, abun-
dance, and health and anthropogenic impacts to
wetlands and uplands, and second, using this infor-
mation to provide amphibian conservation recom-
mendations.
WETLAND TYPE
Depressional wetlands
ASSEMBLAGES
Amphibians
STATUS
PROJECT DESCRIPTION
The King County Wetland-Breeding Amphibian
Monitoring Program was originally designed to pro-
vide the county with long-term wetland, amphibian,
and landscape information for planning and regula-
tory purposes. From 1993 to the present, a mini-
mum of 150 volunteers were trained to census am-
phibian eggs, juveniles, and adults in 90 freshwater
wetlands of 26 watersheds in King County. Analy-
sis of amphibian distribution and health for all wet-
lands was completed. Atargeted subsample of 21
wetlands in 3 rapidly urbanizing watersheds in which
amphibians were declining was additionally moni-
tored for wetland hydrology, predators, and wa-
tershed condition to determine the causes of popu-
lation declines, in hope of stemming declines through
better wetland conservation practices.
Today, a skeleton program totally run by volun-
teers continues to monitor a few of the initially sur-
veyed wetlands without county support. A more
intensive monitoring program targeting amphibians
as bioindicators of wetland condition has been initi-
ated with the cooperation and partnership of the
University of Washington's Certificate Program in
Implementing methods
8O
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Wetland Science and Management, in which sev-
eral students research and monitor an important
wetland and amphibian issue.
The project's monitoring goals include the
following:
Identify the occurrence of the State-endangered
Oregon spotted frog
Determine land uses compatible with wetland
and amphibian conservation objectives
Provide data to help develop and implement
regulations for the protection of amphibians and
their habitats
Identify population distribution status of other
county declining species
Obtain standardized baseline inventory data on
the distribution, abundance, and health of am-
phibians in King County wetlands
Provide information to King County, Washing-
ton State Department of Fish and Wildlife,
Washington State Department of Ecology, and
Federal resource agencies for developing re-
gional wetland and wildlife management pro-
grams
Develop an effective public outreach and edu-
cation program to train citizens to monitor am-
phibians and wetland conditions to foster wet-
land stewardship
Educate potential wetland scientists to the di-
versity of wetland issues and train them in de-
veloping monitoring programs, assessing wet-
land condition, and crafting management plans
for better wetland conservation
Investigate local urban amphibian decline and
extinction issues
Develop methods for utilizing amphibians as
bioindicators of wetland condition by establish-
ing cause-and-effect relationships between
amphibian declines and wetland hydrology,
water quality, predators and pathogens, and
watershed land use.
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WISCONSIN: DEVELOPING BIOLOGICAL INDEXES FOR
WISCONSIN'S PALUSTRINE WETLANDS
Contact
Jennifer Hauxwell
Organization Wi sconsin Department of Natural Resources
Bureau of Integrated Science Services
1350 Femrite Drive
Monona,WI53716
Phone (608)221-6338
E-mail hauxw@dnr.state.wi.us
PURPOSE
Develop a biological index for Wisconsin's
palustrine wetlands
Compare performance of one plant and two
macroinvertebrate multimetric indices
Develop biological integrity rating system for
classifying wetlands
WETLAND TYPE
Palustrine wetlands
ASSEMBLAGES
Macroinvertebrates
Vascular plants
STATUS
Findings representing preliminary investigations are
presented in Final Report to EPA Region V, Wet-
land Grant #CD985491-01-0 prepared by Wis-
consin DNR (January 2000).
PROJECT DESCRIPTION
Field studies for this project were conducted dur-
ing the spring and summer of 1998, with laboratory
analysis and data synthesis completed the following
year. Funding was provided by a grant from EPA
Region 5. The findings formed the basis for a sec-
ond EPA-funded grant to refine and further evalu-
ate the preliminary indices and expand communi-
ties covered to include amphibians, small mammals,
diatoms, and zooplankton.
STUDY DESIGN
We sampled 104 palustrine depression wetlands
distributed across the maj or ecoregions ofWisconsin
during early spring and summer of 1998. Study
sites included a mixture of least-disturbed reference
basins and degraded or restored wetlands, repre-
senting a range in vegetative cover types, water
chemistries, and water duration.
SAMPLING METHODS:
MACROINVERTEBRATES
We sampled macroinvertebrates early in the spring
to minimize influences of immigration-emigration.
Three different field collection procedures were
-------
evaluated and two laboratory approaches were
used. On all 104 wetlands, we collected two sets
of three standard D-frame net sweeps of approxi-
mately 1-m length each. Sampling stations (< 60
cm deep) were established at equally spaced points
around the wetland perimeter that approximately
trisected the basin (and ensured coverage of the
maj or plant communities present). The first set of
net sweeps was concentrated (large coarse materi-
als were rinsed, examined, and removed) into a 1-
quart container; the second set of net sweeps was
not field-concentrated, but rather was placed di-
rectly into a 1-gallon plastic bag. Both sets were
preserved in ethanol and returned to the laboratory
for processing. The Athird set (used on a subset of
17 wetlands) of D-frame net sweep samples was
placed on a coarse wire screen over a collection
basin for a period of approximately 10 minutes.
Organisms falling (or crawling) through the screen
and entering the collection container were collected
and preserved as above. This last set represented
a "clean" sample that was much easier to sort and
process than the standard samples.
LABORATORY METHODS:
MACROINVERTEBRATES
Macroinvertebrates were processed using a two-
tiered approach. The first stage consisted of a fixed
100-count (sensu Hilsenhoff Biotic Index proce-
dures) using a grid-marked tray with 24 cells. Or-
ganisms were picked and sorted at a coarse taxo-
nomic level, usually order or family only. Following
completion of the 100-count sample, we processed
the balance of the sample in its entirety (except for
sub sampling dominant taxa). The unconcentrated
"bag" samples proved to be too large to process in
an economical fashion, so only the complete set of
104 "field-concentrated" samples were processed.
The 17 "screened" samples generally contained less
than 100 total organisms and were processed com-
pletely.
ANALYTICAL METHODS:
MACROINVERTEBRATES
We used SYSTAT to perform all statistical and
graphical analyses. Percentage data were trans-
formed using the arc-sine square-root transforma-
tion, and abundance data were either log-trans-
formed or power-transformed as applicable. Met-
ric development was based on a series of visual
comparisons of community attribute responses to
suspected measures of disturbance using box plots
and jittered dot density plots. Those attributes that
exhibited evidence of separation between reference
wetland conditions and wetlands suspected to be
impacted by human disturbance were selected as
potential metrics. Attributes that exhibited incon-
sistent or overlapping responses between impacted
and reference systems were eliminated from further
consideration.
SAMPLING METHODS: PLANTS
We conducted simplified plant surveys during July
1998 using a combination of techniques. This in-
cluded a subjective estimate of cover and an ob-
jective survey of percent cover and frequency of
occurrence within six equidistantly spaced 20 by
50 cm rectangular quadrats positioned along each
of three transects that trisected the wetland basin
(total of 18 quadrats per wetland).
LABORATORY METHODS: PLANTS
No biomass or stem counts were performed.
Voucher specimens were pressed and identified to
species when possible, but most plant metrics were
based on a coarser taxonomic level.
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ANALYTICAL METHODS: PLANTS
We developed the plant biological index using the
same procedures described for the two
macroinvertebrate-based indices. Because we
wanted to develop a practical tool for managers
with limited botanical expertise, we lumped taxa at
various taxonomic levels (e.g., family, genus) or
structural groups (e.g., grass-like, emergent) for
analysis. Importance values (average of percent
cover and frequency of occurrence) were used as
the attribute of concern for family-genus-species
levels, and percent cover was used for emergent,
submergent, floating-leafed, and open-water
attributes.
OTHER DATA COLLECTED
We also collected associated physical and chemi-
cal data on each wetland, including pH, alkalinity,
conductivity, color, temperature, clarity, and depth.
Riparian cover type within a 100-foot buffer area
surrounding each wetland was subjectively estimated
and recorded, as was shade canopy cover.
PRELIMINARY FINDINGS
Three multimetric indices (two macroinvertebrate
and one plant index) were developed. The Wis-
consin Wetland Macroinvertebrate Index (WWMI)
is a multimetric index based on 15 metrics derived
from a total count of organisms in three composited
net sweeps. Atotal of 69 community attributes were
evaluated. The WWMI is composed of 12 abun-
dance metrics, two richness metrics, and one per-
centage metric. Abundance metrics include mol-
lusks, annelids, fairy shrimp, damselflies, pigmy
backswimmers, water boatmen, limnephild
caddisflies, total caddisflies, phantom midges, mos-
quitoes, soldier flies, and total invertebrates. Rich-
ness metrics are noninsects and total taxa. The
percent caddisflies is the only percentage metric.
Apparent redundancies (e.g., caddisflies) in the
metric may or may not be an issue; differences in
taxonomic rate of development in wetlands due to
thermal dynamics may require a certain amount of
redundancy to ensure that important taxonomic
groups are accounted for. The WWMI is used to
rate, rank, or compare wetland biological condi-
tion.
The second macroinvertebrate index, termed the
100-count macroinvertebrate biotic index (100-
countMBI), is based on 10 metrics derived from a
random pick of 100 organisms found in the three
composited net sweeps. The 100-count MBI is
composed of 9 percentage metrics and 1 richness
metric (noninsecttaxa). Percentage metrics include
pigmy backswimmers, water boatmen, total "bugs,"
limnephilid caddisflies, total caddisflies, chironomids,
soldier flies, and the sum of Ephemeroptera-
Odonata-Trichoptera (EOT) taxa. Noninsects rep-
resent the only richness metric in the 100-count MBI.
The ninth percentage metric, mollusks, may be only
useful in prairie-type wetlands. The 100-count MBI
is intended to be applied in the field by experienced
staff as a means of rapid bioassessment.
The third index is the Wisconsin Wetland Plant
Biotic Index (WWPBI) is based on eight (or nine)
plant metrics derived from transect data (represent-
ing 18 quadrats) and is intended to serve as a supple-
mentary index to the WWMBI to rate, rank, and
compare wetland biological condition. Of 24 plant
community attributes tested, only one richness
(count) metric, one percent metric, and seven im-
portance value-based metrics demonstrated con-
sistent and predictable response. The single rich-
ness metric, total taxa, may require further modifi-
cation after reaching some consistency regarding
taxonomic resolution (currently mixed family-genus-
species). Importance value-based metrics included
Carex, reed canary grass, cattail, duckweed,
bluejoint grass, and "good" plants (the sum of a
group of plants including all Carex, Utricularia,
Potamogeton, Calamogrostis, Sagittaria,
Polygonum, andEquisetum). An additional impor-
tance value-based metric, "pondweeds," would only
be applied to wetlands with water duration exceed-
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ing 7 months per year. The only percentage-based
metric, floating-leafed plants, would likewise only
be applied to wetlands with water durations ex-
ceeding 7 months.
LESSONS LEARNED
Macroinvertebrates
Water duration is an important factor shaping
macroinvertebrate community composition and
derived metrics that must be accounted for in
metric scoring.
A coarse level of taxonomic resolution (order
and family) appears to be satisfactory in devel-
oping wetland macroinvertebrate metrics.
Issues relating to redundancy among metrics,
influences of water chemistry, differences among
ecoregions, and seasonal variations need to be
addressed in more detail. Undoubtedly, these
factors need to be accounted for in establishing
rating scores and/or in refining metrics for use
in different areas or habitats.
Basic differences exist in macroinvertebrate
communities between wetlands representing
wooded kettle depressions and open-prairie
type depressions in Wisconsin.
Our greatest difficulty was in selecting and as-
signing some measure of "human impact" to the
study site wetlands. Further research will be
required to quantify the degree of human im-
pact in order to refine biological response
metrics and indices.
The WWMBI was not stable across dates (and
not designed to be); consequently, its use is re-
stricted to early spring.
Each macroinvertebrate index has its own set
of advantages and disadvantages; further re-
finement is required to enable their successful
application in the field.
Plants
Some biases were apparent in the WWPBI, as
reference kettles scored consistently higher than
prairie wetlands.
WWPBI scores in restored prairie wetlands
were better than in many natural wetlands, sug-
gesting that wetland restorations in Wisconsin
may be adequate in terms of "restoring" the
vegetative community (not true for
macroinvertebrate response).
The WWPBI shows some promise in its per-
formance and, because of its taxonomic sim-
plicity, it could be applied by nonbotanists.
FOLLOWUP: CONTINUING WORK
With the assistance of a second EPA wetland grant,
we have refined the macroinvertebrate and plant
multimetric indices and explored expanding the com-
munity components to include zooplankton (in co-
operation with Dr. Stanley Dodson, University of
Wisconsin-Madison), diatoms (Paul Garrison, Wis-
consin DNR), amphibians, and small mammals.
Findings associated with the second grant are re-
ported in the second Wisconsin case study below.
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WISCONSIN: REFINEMENT AND EXPANSION OF THE
WISCONSIN WETLAND BIOLOGICAL INDEX
FOR ASSESSMENT OF DEPRESSIONAL
PALUSTRINE WETLANDS
Contact
Jennifer Hauxwell
Organization Wi sconsin Department of Natural Resources
Bureau of Integrated Science Services
1350 Femrite Drive
Monona,WI 53716
Phone (608)-221-6373
E-mail jennifer.hauxwell@dnr.state.wi.us
PURPOSE
Test and refine a Biotic Index for Wisconsin's
palustrine wetlands
Expand list of assemblages to include
macroinvertebrates, zooplankton, diatoms,
amphibians, plants, and small mammals
Establish a biological integrity rating system for
classifying wetlands based on the response of
selected biological attributes (metrics) of the
above communities to surrogate measures of
human disturbance
WETLAND TYPE
Palustrine wetlands
ASSEMBLAGES
Macroinvertebrates
Aquatic plants
Zooplankton
Diatoms
Amphibians
Small mammals
STATUS
Findings presented in Final Report to EPARe-
gion V prepared by Wisconsin DNR dated April
2002.
PROJECT DESCRIPTION
This proj ect represents the evaluation and expan-
sion phase of an earlier study that resulted in the
preliminary development of a Wisconsin Wetland
Biological Index based on plant and
macroinvertebrate metrics (please see final report
to EPA -Wetland Grant #CD985491 -01 -0). Data
from this second study were used to refine and fur-
ther evaluate the preliminary indices and expand
communities covered to include zooplankton, dia-
toms, amphibians, and small mammals. Field stud-
ies for thi s proj ect were conducted during the spring
and summer of 2000, with laboratory analysis and
data synthesis completed in 2001. Funding was
provided by a grant from EPA Region 5.
We sampled 75 palustrine depression wetlands in
southeast Wisconsin during early spring and sum-
mer of 2000. Study sites included a mixture of least-
disturbed reference basins (17 prairie and 19
86
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wooded kettles) and impacted wetlands (18 urban
and 20 agriculture), representing a range in vegeta-
tive cover types and water chemistries. A severe
drought, which began the previous year, caused most
of the smaller wetlands with short hydroperiods
(seasonal and temporary) to dry out, and conse-
quently only long-duration wetlands (semiperma-
nent and permanent) were sampled.
We sampled macroinvertebrates early in the spring
to minimize influences of immigration-emigration.
On each wetland, we collected three standard D-
frame net sweeps of approximately 1-m length each.
Sampling stations (less than 60 cm deep) were es-
tablished at equally spaced points around the wet-
land perimeter that approximately trisected the ba-
sin and assured coverage of the maj or plant com-
munities present. The contents of the three net
sweeps were concentrated (large coarse materials
were rinsed, examined, and removed) into a 1 -quart
container, preserved in ethanol, and returned to the
laboratory for processing. Seven field replicates
were collected.
LABORATORY METHODS:
MACROINVERTEBRATES
Each macroinvertebrate sample was processed
entirely, using a two-phase approach with
subsampling employed only for extremely abundant
organisms. The first stage involved using a grid-
marked tray with 24 cells. Cells were selected ran-
domly, and organisms were picked and sorted at a
coarse taxonomic level, usually to order or family
level. The abundance of organisms present in the
first two or three randomly selected cells was used
to proj ect the abundance of the most common taxa
in the sample. Taxa whose abundance was esti-
mated to exceed 3 00 in the total sample were then
overlooked while the balance of the sample was
processed (the second phase of processing). All
specimens were vouchered (preserved in 70% etha-
nol) for possible further evaluation.
SAMPLING METHODS: PLANTS
We conducted simplified plant surveys during July
2000 using a combination of techniques. This in-
cluded a subjective estimate of cover and an ob-
jective survey of percent cover and frequency of
occurrence within six equidistantly spaced 20x50-
cm rectangular quadrats positioned along each of
three transects that trisected the wetland basin (to-
tal of 18 quadrats per wetland).
LABORATORY METHODS: PLANTS
No biomass or stem counts were performed.
Voucher specimens were pressed and identified to
species when possible, but most plant metrics are
based on a coarser taxonomic level.
SAMPLING METHODS: ZOOPLANKTON
Note: zooplankton studies were conducted by Dr.
Stanley Dodson, University of Wisconsin-Madison,
Zoology Department, Madison, Wisconsin. E-mail:
sidodson@facstaff.wis.edu.
We collected one zooplankton sample from a cen-
tral basin location in each wetland during June 2000
using a 5-L plastic bucket. We filtered a known
volume of water through a No. 10 (60-micron mesh)
net to capture zooplankton within. Seven field rep-
licates were collected. Samples were preserved in
70% ethanol until processed.
LABORATORY METHODS:
ZOOPLANKTON
Each sample was scanned at moderate magnifi-
cation for species of cladocera, copepods, ostra-
cods, and aquatic insects. Slides and dissections
were made where necessary; for example, to aid in
the identification of copepod species. Total num-
ber of male and female Daphnia was counted in
87
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each sample. More than 200 slides were prepared,
and identification of organisms is in progress. Or-
ganisms will be identified to species where possible.
SAMPLING METHODS: DIATOMS
Diatom studies were conducted under the direc-
tion of Paul Garrison, WDNR, Bureau of Integrated
Science Services, Environmental Contaminants
Section, 1350FemriteDrive,Monona,WI 53716.
E-mail: garrip@dnr. state, wi.us
Using a 1-dram vial as a sample collection de-
vice, we collected and composited surficial (upper
0-1 cm) sediments from five sites in each wetland.
Seven field replicates were collected. Samples were
kept on ice or refrigerated until processed.
LABORATORY METHODS: DIATOMS
Each diatom sample will be thoroughly mixed, and
a small amount will be placed into a tall beaker.
Hydrogen peroxide will be added and the sample
will be allowed to steep for about 5 minutes. Po-
tassium dichromate will be added (under a venti-
lated hood and handled with safety gloves) to fa-
cilitate reduction of organic matter. The sample will
be washed at least four times with deionized water
by centrifuging for 10 minutes. Two portions of the
cleaned sample will be dried on separate No. 1
cover-slips and mounted with Naphraxa, and la-
beled accordingly. Specimens from both cover slips
will be identified and counted under oil immersion
objective (1,400X) until a total of 250 frustules are
counted. Identification of difficult taxa will be made
using a scanning electron microscope. Undeter-
mined specimens representing a significant portion
of a sample will be sent to Dr. Rex Lowe at Bowl-
ing Green University, Dr. Jan Stevenson at Louis-
ville University, and/or Dr. Gene Stormer at the
University of Michigan.
SAMPLING METHODS: AMPHIBIANS
Because amphibians are extremely sensitive to
weather and temperature, we assessed amphibian
communities during two separate sampling periods.
We conducted standardized frog-toad calling sur-
veys (using WDNR protocols) during the first two
phenologies (early spring and late spring) between
the hours of 8 and 10:30 pm for 10-15 minutes
when water temperatures were above 50°F during
the first phenology or above 60°F during the sec-
ond phenology. We recorded all calling to permit
verification of questionable identifications. In addi-
tion to calling, we added to the database any per-
sonal observations of amphibians made during any
of the daylight visits and any specimens captured
during the macroinvertebrate surveys.
LABORATORY METHODS: AMPHIBIANS
No laboratory methods were used.
SAMPLING METHODS:
SMALL MAMMALS
Small mammal studies were conducted by R.
Bautz, WDNR, Bureau of Integrated Science Ser-
vices, Ecological Inventory & Monitoring Section,
1350 Femrite Drive, Monona, WI 53716. E-mail:
bautzr@dnr. state, wi .us.
We assessed small mammal communities by trap-
ping during August-September 2000. On each
wetland, we set 46 baited traps (mixture of 40 mu-
seum special grade and 6 larger tomahawk traps)
along transects (zigzag scattered routes) in the ri-
parian zone (variable dimensions, depending on
setting) for one night. A one-day/night trapping
period was used to minimize disturbance by rac-
coons and other predators. Bait consisted of a mix-
ture of peanut butter and rolled oats. Traps were
cleaned and rebaited each morning. Specimens
were placed in labeled freezer bags and returned to
the laboratory for identification.
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LABORATORY METHODS:
SMALL MAMMALS
Identifications of rare taxa and species of special
concern were verified by local experts from the
University of Wisconsin.
OTHER DATA COLLECTED
We also collected associated physical and chemi-
cal data on each wetland. The water chemistry
measures included pH, alkalinity, conductivity, color,
chloride, calcium, silica, nitrate-nitrite, organic-ni-
trogen, and total phosphorus. Chemical analyses
were performed at the Wisconsin State Laboratory
of Hygiene following standard EPA-approved pro-
cedures. Physical data collected included water
depth, apparent color, temperature, and size. Ri-
parian cover type within a 100-foot buffer area sur-
rounding each wetland was subjectively estimated
and recorded, as well as shade canopy cover.
ANALYTICAL METHODS: DATA
ANALYSIS AND DEVELOPMENT OF
BIOLOGICAL INDICES
Data were entered into Excel spreadsheet and
Access databases and examined for entry errors
(univariate checks) and anomalies (e.g., bivariate
plots) prior to analysis. Various community or spe-
cies attributes (e.g., taxa or species richness, diver-
sity, presence or absence of selected functional feed-
ing guilds, trophic structure, percentages) were
evaluated and scored as potential metrics based on
their responsiveness to measures of human distur-
bance. Community attributes were examined using
a combination of procedures (outlined below) to
select promising metrics for index development.
Attributes that exhibited strong positive or negative
correlation with selected human response variables
were selected as metrics. We compared the sensi-
tivity and correspondence among the selected com-
munity metrics, and developed a single multimetric
index of ecosystem integrity that related to overall
wetland condition.
Because different forms of human disturbance
elicit different responses among the various wet-
land biological communities, it is difficult if not im-
possible to choose one measure that represents "the"
single best measure of human disturbance. For ex-
ample, the amphibian community may respond more
directly to woodland impacts (distance to nearest
woodland, patch size, corridor dimensions, etc.)
than to nutrient or pesticide inputs, whereas zoop-
lankton and diatom communities may be more re-
sponsive to percent row crops in the watershed.
Consequently, we evaluated biotic responses to a
combination of a priori classes of human disturbance
and a composite chemical index that incorporated
nitrogen, phosphorus, and chloride concentrations.
The a priori classes represented agricultural and
urban impacts as well as subclasses of each, in-
cluding three intensity levels of agricultural impact
and two forms of urban impact. Biotic responses
occurring within these classes were measured
against the responses in least disturbed kettles and
prairie depression wetlands. Statistical procedures
and approaches used in the analysis varied among
the communities assessed because of inherent dif-
ferences in responses and the type of data collected.
Canonical correspondence analysis was used to
explore unimodal distributions of the diatoms along
the various environmental gradients. We used
SYSTAT and a combination of other available sta-
tistical software programs to perform statistical and
graphical analyses. Metric development was based
on a series of visual comparisons of community at-
tribute responses to measures of disturbance (com-
bination of selected water chemistry and land use
characteristics as discussed above) using box plots
and jittered dot density plots. Attributes that ex-
hibited evidence of separation between reference
and impacted wetlands were selected as metrics
(or incorporated into the existing list of metrics).
Attributes that exhibited inconsistent or overlapping
responses between impacted and reference systems
were eliminated from further consideration.
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LESSONS LEARNED
Finding willing and cooperative private prop-
erty landowners for access to potentially im-
pacted wetlands is difficult!
Determining what attribute or attributes (e.g.,
land use, chemical contaminant concentration,
distance to nearest road) to use as surrogate
measures of human disturbance is critical to the
successful development of reliable multimetric
biological indices.
Drought (or floods) can seriously hamper sam-
pling plans and interfere with the best of plans!
The list of suitable metrics among macroinver-
tebrate communities of permanent wetlands is
different and much shorter than the list of metrics
suitable for wetlands with shorter water dura-
tion periods.
9O
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REFERENCES
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BorthC. 1997. Development of vegetation indices
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Burton TM, Uzarski DQ Gathman JP, Genet JA,
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GLOSSARY
Aquatic life use A type of designated use pertain-
ing to the support and maintenance of healthy bio-
logical communities.
Assemblage An association of interacting popu-
lations of organisms that belong to the same maj or
taxonomic groups. Examples of assemblages used
forbioassessments include: algae, amphibians, birds,
fish, amphibians, macroinvertebrates (insects, cray-
fish, clams, snails, etc.), and vascular plants.
Attribute Ameasurable component of a biologi-
cal system. In the context of bioassessments, at-
tributes include the ecological processes or char-
acteristics of an individual or assemblage of species
that are expected, but not empirically shown, to
respond to a gradient of human disturbance.
Benthos The bottom fauna of waterbodies.
Biological assessment (bioassessment) Using
biomonitoring data of samples of living organisms
to evaluate the condition or health of a place (e.g.,
a stream, wetland, or woodlot).
Biological integrity "the ability of an aquatic eco-
system to support and maintain a balanced, adap-
tive community of organisms having a species com-
position, diversity, and functional organization com-
parable to that of natural habitats within a region"
(Karr and Dudley 1981).
Biological monitoring Sampling the biota of a
place (e.g., a stream, a woodlot, or a wetland).
Biota All the plants and animals inhabiting an area.
Composition (structure) The composition of the
taxonomic grouping such as fish, algae, or
macroinvertebrates relating primarily to the kinds
and number of organisms in the group.
Community All the groups of organisms living to-
gether in the same area, usually interacting or de-
pending on each other for existence.
Competition Utilization by different species of lim-
ited resources of food or nutrients, refugia, space,
ovipositioning sites, or other resources necessary
for reproduction, growth, and survival.
Criteria A part of water quality standards. Crite-
ria are the narrative and numeric definitions condi-
tions that must be protected and maintained to sup-
port a designated use.
Continuum A gradient of change.
Designated use Apart of water quality standards.
A designated use is the ecological goal that
policymakers set for a waterbody, such as aquatic
life use support, fishing, swimming, or drinking wa-
ter.
Disturbance "Any discrete event in time that dis-
rupts ecosystems, communities, or population struc-
ture and changes resources, substrate availability
or the physical environment" (Picket and White
1985). Examples of natural disturbances are fire,
drought, and floods. Human-caused disturbances
are referred to as "human disturbance" and tend to
be more persistent over time, e.g., plowing,
clearcutting of forests, conducting urban stormwater
into wetlands.
Diversity A combination of the number of taxa
(see taxa richness) and the relative abundance of
those taxa. A variety of diversity indexes have been
developed to calculate diversity.
Dominance The relative increase in the abundance
of one or more species in relation to the abundance
of other species in samples from a habitat.
Ecological risk assessment An evaluation of the
potential adverse effects that human activities have
on the plants and animals that make up ecosystems.
Ecosystem Any unit that includes all the organ-
isms that function together in a given area interact-
ing with the physical environment so that a flow of
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energy leads to clearly defined biotic structure and
cycling of materials between living and nonliving
parts (Odum 1983).
Ecoregion Aregion defined by similarity of cli-
mate, landform, soil, potential natural vegetation,
hydrology, and other ecologically relevant variables.
Gradient of human disturbance The relative
ranking of sample sites within a regional wetland
class based on degrees of human disturbance (e.g.,
pollution, physical alteration of habitats, etc.)
Habitat The sum of the physical, chemical, and
biological environment occupied by individuals of a
particular species, population, or community.
Hydrology The science of dealing with the prop-
erties, distribution, and circulation of water both on
the surface and under the earth.
Impact Achange in the chemical, physical (includ-
ing habitat), or biological quality or condition of a
waterbody caused by external forces.
Impairment Adverse changes occurring to an eco-
system or habitat. An impaired wetland has some
degree of human disturbance affecting it.
Index of biologic integrity (IBI) An integrative
expression of the biological condition that is com-
posed of multiple metrics. Similar to economic in-
dexes used for expressing the condition of the
economy.
Intolerant taxa Taxa that tend to decrease in wet-
lands or other habitats that have higher levels of
human disturbances, such as chemical pollution or
siltation.
Macroinvertebrates Animals without backbones
(insects, crayfish, clams, snails, etc.) that are caught
with a 500-800 micron mesh net.
Macroinvertebrates do not include zooplankton or
ostracods, which are generally smaller than 200 mi-
crons in size.
Metric An attribute with empirical change in value
along a gradient of human disturbance.
Minimally impaired site Sample sites within a
regional wetland class that exhibit the least degree
of detrimental effect. Such sites help anchor gradi-
ents of human disturbance and are commonly re-
ferred to as reference sites.
Most-impaired site Sample sites within a regional
wetland class that exhibit the greatest degree of
detrimental effect. Such sites help anchor gradients
of human disturbance and serve as important refer-
ences, although they are not typically referred to as
reference sites.
Population A set of organisms belonging to the
same species and occupying a particular area at the
same time.
Reference site (as used with an index of bio-
logical integrity) A minimally impaired site that is
representative of the expected ecological conditions
and integrity of other sites of the same type and
region.
Stressor Any physical, chemical, or biological en-
tity that can induce an adverse response.
Taxa A grouping of organisms given a formal taxo-
nomic name such as species, genus, family, etc. The
singular form is taxon.
Taxa richness The number of distinct species or
taxa that are found in an assemblage, community,
or sample.
Tolerance The biological ability of different spe-
cies or populations to survive successfully within a
certain range of environmental conditions.
Trophic Feeding, thus pertaining to energy
transfers.
Wetland(s) (1) Those areas that are inundated or
saturated by surface or groundwater at a frequency
and duration sufficient to support, and that under
normal circumstances do support, a prevalence of
vegetation typically adapted for life in saturated soil
conditions [EPA, 40 C.RR.§ 230.3 (t)/USAGE,
33 C.RR. § 328.3 (b)]. (2) Wetlands are lands
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transitional between terrestrial and aquatic systems
where the water table is usually at or near the sur-
face or the land is covered by shallow water. For
the purposes of this classification, wetlands must
have one or more of the following three attributes:
(a) at least periodically, the land supports predomi-
nantly hydrophytes, (b) the substrate is predomi-
nantly undrained hydric soil, and (c) the substrate is
nonsoil and is saturated with water or covered by
shallow water at some time during the growing sea-
son of each year (Cowardinetal. 1979). (3) The
term "wetland" except when such term is part of
the term "converted wetland," means land that (a)
has a predominance of hydric soils, (b) is inundated
or saturated by surface or ground water at a fre-
quency and duration sufficient to support a preva-
lence of hydrophyte vegetation typically adapted
for life in saturated soil conditions, and (c) under
normal circumstances does support a prevalence
of such vegetation. For purposes of this Act and
any other Act, this term shall not include lands in
Alaska identified as having a high potential for agri-
cultural development which have a predominance
of permafrost soils [Food Security Act, 16 U.S.C.
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