United States           Office of Water
Environmental Protection Agency   4304T
June 2008
Nutrient Criteria
Technical Guidance Manual


U.S. Environmental Protection Agency

          Nutrient Criteria
     Technical Guidance Manual

            June 2008


This manual provides technical guidance to States, Authorized Tribes, Territories and other authorized
jurisdictions to establish water quality criteria and standards under the Clean Water Act (CWA), in order
to protect aquatic life from acute and chronic effects of nutrient over-enrichment. Under the CWA, States
and Authorized Tribes are directed to establish water quality criteria to protect designated uses. States
and Authorized Tribes may use approaches for establishing water quality criteria that differ from those
recommended in this guidance. This manual constitutes EPA's scientific recommendations regarding the
development of numeric criteria reflecting ambient concentrations of nutrients that protect aquatic life.
However, it does not substitute for the CWA or EPA's regulations; nor is it a regulation itself. Thus, it cannot
impose legally binding requirements on EPA, States, Authorized Tribes, or the regulated community, and
might not apply to a particular situation or circumstance. Further, States and Authorized Tribes may choose to
develop different types of criteria for wetlands protection, including narrative criteria. EPA may change this
guidance in the future.
                                         Nutrient Criteria-Wetlands


Disclaimer     	i
List of Figures  	iv
List of Tables   	iv
List of Internet Link References	v
Contributors    	vii
Acknowledgments     	vii
Executive Summary    	ix

CHAPTER 1.     Introduction    	1-1

1.1      Introduction    	1-1
1.2      Water Quality Standards and Criteria    	1-2
1.3      Nutrient Enrichment Problems  	1-3
1.4      Overview of the Criteria Development Process   	1-6
1.5      Roadmap to the Document     	1-6

CHAPTER 2.     Overview of Wetland Science    	2-1

2.1      Introduction    	2-1
2.2      Components of Wetlands       	2-2
2.3      Wetland Nutrient Components  	2-5

CHAPTER 3.     Classification of Wetlands       	3-1

3.1      Introduction    	3-1
3.2      Existing Wetland Classification Schemes        	3-2
3.3      Sources of Information for Mapping Wetland Classes    	3-10
3.4      Differences in Nutrient References Condition or Sensitivity to
        Nutrients Among Wetland Classes	3-10
3.5      Recommendations      	3-11

CHAPTER 4.     Sampling Design for Wetland Monitoring	4-1

4.1      Introduction    	4-1
4.2      Considerations for Sampling Design    	4-2
4.3      Sampling Protocol      	4-5
4.4      Summary      	4-9

CHAPTER 5.     Candidate Variables for Establishing Nutrient Criteria   	5-1

5.1      Overview of Candidate Variables       	5-1
5.2      Supporting Variables   	5-3
5.3      Casual Variables        	5-5
5.4      Response Variables     	5-9
5.5      Summary      	5-12
                                 Nutrient Criteria-Wetlands

                                  CONTENTS (continued)

CHAPTER 6.     Database Development and New Data Collection  	6-1

6.1      Introduction    	6-1
6.2     Databases and Database Management   	6-1
6.3     Quality of Historical and Collected Data	6-4
6.4     Collecting New Data    	6-6
6.5     Quality Assurance / Quality Control    	6-7

CHAPTER?.     Data Analysis   	7-1

7.1      Introduction   	7-1
7.2     Factors Affecting Analysis Approach   	7-1
7.3      Distribution-based Approaches  	7-2
7.4      Response-based Approaches     	7-3
7.5      Partitioning Effects Among Multiple Stressors  	7-5
7.6      Statistical Techniques   	7-5
7.7      Linking Nutrient Availability to Primary Producer Response    	7-8

CHAPTERS.     Criteria Development    	8-1

8.1      Introduction    	8-1
8.2     Methods for Developing Nutrient Criteria       	8-1
8.3     Evaluation of Proposed Criteria 	8-6

References     	R-l
Supplementary References      	R-21
Appendix A. Acronym List and Glossary       	A-l
Acronyms     	A-l
Glossary       	A-2
Appendix B. Case Study: Deriving a Phosphorus Criterion for the Florida Everglades    	B-l
Literature Cited	L-l
                                     Nutrient Criteria-Wetlands                                    iii

                                           LIST OF FIGURES

FIGURE 1.1      Flowchart providing the steps of the process to develop wetland nutrient criteria          	1-8
FIGURE 2.1      Schematic of nutrient transfer among potential system sources and sinks                 	2-1
FIGURE 2.2      Relationship between water source and wetland vegetation. Modified from Brinson (1993)       	2-2
FIGURE 2.3      Schematic showing basic nutrient cycles in soil-water column of a wetland     	2-7
FIGURE 2.4      Range of redox potentials in wetland soils   	2-8
FIGURE 2.5      Schematic of the nitrogen cycle in wetlands 	2-9
FIGURE 2.6      Schematic of the phosphorus cycle in wetlands       	2-10
FIGURE 3.1      Map of Omernik aquatic ecoregions        	3-3
FIGURE 3.2A     Map of Bailey ecoregions with coastal and estuarine provinces       	3-3
FIGURE 3.2B     Legend (of Map of Bailey ecoregions with coastal and estuarine provinces)    	3-4
FIGURE 3.3      Examples of first four hierarchical levels of Ecological Units   	3-5
FIGURE 3.4A     Cowardin hierarchy of habitat types for estuarine systems     	3-6
FIGURE 3.4B     Cowardin hierarchy of habitat types for Palustrine systems    	3-7
FIGURE 3.5      Dominant water sources to wetlands, from Brinson  1993      	3-8
FIGURE 3.6      Dominant hydrodynamic regimes for wetlands based on flow pattern   	3-8
FIGURE 3.7      Interaction with break in slope with groundwater inputs to slope wetlands (Bottom)
                Palustrine systems, from Cowardin et al. 1979       	3-8
FIGURE 5.1      Conceptual model of causal pathway between human activities and ecological attributes 	5-2
FIGURE 7.1      Biological condition gradient model describing biotic community condition as levels of
                stressors increase	7-1
FIGURE 8.1      Use of undisturbed wetlands as a reference for establishing criteria versus an effects-based
                approach         	8-3
FIGURE 8.2      Tiered aquatic life use model used in Maine 	8-4
FIGURE 8.3      Percent calcareous algal mat cover in relation to distance from the P source showing the loss
                of the calcareous algal mat in those sites closer to the source   	8-5

                                            LIST OF TABLES

TABLE 1         Observed consequences of cultural eutrophication in freshwater wetlands      	1-5

TABLE 2         Comparison of landscape and wetland classification schemes  	3-12

TABLE 3         Features of the major hydrogeomorphic classes of wetlands that may influence background nutrient
                concentrations, sensitivity to nutrient loading, nutrient storage forms and assimilative capacity,
                designated use and choice of endpoints     	3-13
Comparison of Stratified Probabilistic, Targeted, and BACI Sampling Designs   	4-10
                                           Nutrient Criteria-Wetlands



EXECUTIVE SUMMARY    http://www.epa.gov/waterscience/criteria/nutrient/guidance/index.html
http ://www. epa. gov/owow/wetlands/bawwg/case/or.html
http ://www. epa. gov/waterscience/criteria/wetlands/1 TLandUse .pdf
http ://www. epa. gov/owow/wetlands/bawwg/case/mtdev.html
http ://www. epa. gov/owow/wetlands/bawwg/case/fl 1 .html
http ://www. epa. gov/owow/wetlands/bawwg/case/fl2 .html
http ://www. epa. gov/owow/wetlands/bawwg/case/oh 1 .html
http ://www. epa. gov/waterscience/criteria/wetlands/
http ://www. epa. gov/waterscience/criteria/wetlands/16Indicators
                                         Nutrient Criteria-Wetlands

http ://www. epa.gov/emap
http ://water.usgs .gov/nrp/webb/about.html
http ://www.usbr.gov
http: //el. erdc .usac e. army.mil/wrap/wrap .html
http ://www. stat.uiowa. edu

http://data.lca.gov/Ivan6/app/app _c_ch9.pdf
http ://www. epa.gov/owow

http ://www. dep. state. fl .us/lab s/library/index .htm
http ://www. wwwalker.net/dmsta


http ://www.www. sfwmd.gov
                                          Nutrient Criteria-Wetlands

Nancy Andrews (U.S. Environmental Protection Agency)
Mark Clark (University of Florida)
Christopher Craft (University of Indiana)
William Crumpton (Iowa State University)
Ifeyinwa Davis (U.S. Environmental Protection Agency)
Naomi Detenbeck (U.S. Environmental Protection Agency)
Paul McCormick (U.S. Geological Survey)
Amanda Parker (U.S. Environmental Protection Agency)
Kristine Pintado (Louisiana Department of Environmental Quality)
Todd Rasmussen (University of Georgia)
Ramesh Reddy (University of Florida)
R. Jan Stevenson (Michigan State University)
Arnold van der Valk (Iowa State University)
The authors wish to acknowledge the efforts and input of several individuals. These include members of our
EPA National Nutrient Team: Jim Carleton, Lisa Larimer, Steve Potts, and Robert Cantilli; members of the
EPA Wetlands Division: Kathy Hurld, Chris Faulkner and Donna Downing; members of the Office of Research
and Development and their Office of Science Policy: Valerie Chan, Joseph Schubauer-Berigan, Charles Lane,
John Morrice, Christine Weilhoefer, Tony Olsen, and Walt Nelson; the Office of General Counsel: Leslie
Darman. We also want to thank Kristine Pintado (DEQ, Louisiana) for her contributions and her careful
review and comments.

This document was peer reviewed by a panel of expert scientists. The peer review charge focused on
evaluating the scientific  validity of the processes and techniques for developing nutrient criteria described in
the guidance. The peer review panel comprised Dr. Robert H. Kadlec, Dr. Lawrence Richards Pomeroy, Dr.
Eliska Rejmankova and  Dr. Li Zhang. Edits and suggestions made by the peer review panel were incorporated
into the final version of the guidance.
                                       Nutrient Criteria-Wetlands                                     vii

viii                                       Nutrient Criteria-Wetlands

                                    EXECUTIVE SUMMARY

The purpose of this document is to provide scientifically defensible guidance to assist States, Authorized
Tribes, Territories, and other authorized jurisdictions—hereafter referred to as States—in assessing the
nutrient status of their wetlands, and to provide technical assistance for developing numeric nutrient criteria
for wetland systems in an eco-region.  The development of nutrient criteria is part of an initiative by the U.S.
Environmental Protection Agency (USEPA) to address the problem of cultural eutrophication, i.e., nutrient
pollution caused by human activities (USEPA 1998a).  Cultural eutrophication is not new; however, traditional
efforts at nutrient control have been only moderately successful. Specifically, efforts to control nutrients
in water bodies that have multiple nutrient sources (point and nonpoint sources) have been less effective
in providing satisfactory, timely remedies for enrichment-related problems. Development and adoption
of numeric criteria into water quality standards aids nitrogen and phosphorus pollution control efforts by
providing clear numeric goals for water quality protection. Furthermore, numeric nutrient criteria provide
specific water quality goals that will assist researchers in designing improved best management practices.

In 1998, the USEPA published a report entitled, National Strategy for the Development of Regional Nutrient
Criteria (USEPA 1998a). This report outlines a framework for development of waterbody-specific technical
guidance that can be used to assess nutrient status and develop region-specific numeric nutrient criteria. The
document presented here is the wetland-specific technical guidance  for developing numeric nutrient criteria.
The Nutrient Criteria Technical Guidance Manuals for Rivers and Streams (USEPA, 2000b), Lakes and
Reservoirs (USEPA, 2000a) and Estuarine and Coastal Marine Waters (USEPA, 2001) have been completed
and are available at: http://www.epa.gov/waterscience/criteria/nutrient/guidance/index.html.

Section 303(c) of the Clean Water Act directs States to adopt water quality standards for waters that are
"waters of the United States," including wetlands that are waters of the United States . A water quality
standard consists of three main elements: (1) one or more designated uses of each of the State's waters, such as
recreation or propagation offish; (2) criteria expressed as pollutant concentration levels or narrative statements
representing a quality of water that supports a designated use; and, (3) an anti-degradation policy to protect
existing uses and high quality waters.

The information used in developing the technical approaches in this document came from references  about
studies of wetlands  in a wide range of conditions, but not wetlands with a high degree of modification (e.g.,
wetlands that are considered "prior converted cropland" or artificial wetlands specifically engineered to protect
or improve downstream water quality). This guidance is to assist States in developing numeric nutrient criteria
for wetlands, should they choose to do so. States  may choose to develop different types of criteria for wetlands
protection, including site-specific or narrative criteria for wetlands protection, as long as they are scientifically
defensible and protective of the designated uses (40 CFR § 131.11). This Guidance Manual includes chapters
dealing with the following topics:
1For further information regarding the scope of 'waters of the U.S.' in light of the U.S. Supreme Court's 2006 decision in Rapanos v.
United States, see "Clean Water Act Jurisdiction Following the U.S. Supreme Court's Decision in Rapanos v. United States & Carabell v.
United States," which was jointly issued by the U.S. Environmental Protection Agency and the Army Corps of Engineers and is available
at: http://www.epa.gov/owow/wetlands/.
                                         Nutrient Criteria-Wetlands                                        ix


Classification strategies for nutrient criteria development include:

        • physiographic regions
        • hydrogeomorphic class
        • water depth and duration
        • vegetation type or zone

Choosing a specific classification scheme will likely depend on practical considerations, such as: whether a
classification scheme is available in mapped digital form or can be readily derived from existing map layers;
whether a hydrogeomorphic or other classification scheme has been refined for a particular region and wetland
type; and, whether classification schemes are already in use for monitoring and assessment of other waterbody
types in a State or region.


Three sampling designs for new wetland monitoring programs are described:

        • probabilistic sampling
        • targeted/tiered approach
        • BACI (Before/After, Control/Impact)

These approaches are designed to obtain a significant amount of information for statistical analyses
with relatively minimal effort. Sampling efforts should be designed to collect information that will
answer management questions in a way that permits robust statistical analysis. In addition, site selection,
characterization of reference sites or systems, and identification of appropriate index periods are all of
particular concern when selecting an appropriate  sampling design.  Careful selection of sampling design will
allow the best use of financial resources and  result in the collection of high quality data for evaluation of the
wetland resources of a  State.


Candidate variables to use in determining nutrient condition of wetlands and to help identify appropriate
nutrient criteria for wetlands consist of supporting variables, causal variables,  and response variables.
Supporting variables provide information useful in normalizing causal and response variables and categorizing
wetlands. Causal variables are intended to characterize nutrient availability (or assimilation) in wetlands
and could include nutrient loading rates and soil nutrient concentrations. Response variables are intended to
characterize biotic response and could include community structure and composition of macrophytes and
algae. Recommended variables for wetland nutrient criteria development described in this chapter are:

        1.  Causal variables - nutrient loading rates, land use, extractable and total soil nitrogen (N) and
           phosphorus (P), water column N and P;
        2.  Response variables - nutrient content of wetland vegetation (algal  and/or higher plants), above
           ground biomass and stem height, macrophyte, algal, and macroinvertebrate community structure
           and composition; and,
        3.  Supporting variables - hydrologic condition balance, conductivity, soil pH, soil bulk density, soil
           organic matter content.
                                     Nutrient Criteria-Wetlands


A database of relevant water quality information can be an invaluable tool to States as they develop nutrient
criteria. In some cases, existing data are available and can provide additional information that is specific to
the region where criteria are to be set. However, little or no data are available for most regions or parameters,
and creating a database of newly gathered data is strongly recommended. In the case of existing data, the data
should be geolocated, and their suitability (type and quality and sufficient associated meta data) ascertained.


Data analysis is critical to nutrient criteria development. Proper analysis and interpretation of data determine
the scientific defensibility and effectiveness of the criteria. Therefore, it is important to evaluate short and
long-term goals for wetlands of a given class within the region of concern. The purpose of this chapter is to
explore methods for analyzing data that can be used to develop nutrient criteria consistent with these goals.
Techniques discussed in this chapter include:

        • Distribution-based approaches that examine distributions of primary and supporting
          variables (i.e., the percentile approach);
        • Response-based approaches that develop relationships between measurements of nutrient
          exposure and ecological responses (i.e., tiered aquatic life uses);
        • Partitioning effects of multiple stressors;
        • Statistical techniques;
        • Multi-metric indices; and,
        • Linking nutrient availability to primary  producer response.


Several methods can be used to develop numeric nutrient criteria for wetlands. They include, but are not
limited to, criteria development methods that are detailed in this document:

        • Comparing conditions in known reference systems for each established wetland type and class
          based on best professional judgment or identifying reference conditions using frequency
          distributions of empirical data and identifying criteria using percentile selections of data plotted
          as frequency distributions;
        • Refining classification systems using models, and/or examining system biological attributes in
          comparison to known reference conditions to assess the relationships among nutrients, vegetation
          or algae, soil, and other variables and identifying criteria based on thresholds where those
          response relationships change; and,
        • Using or modifying published nutrient and vegetation, algal, and soil relationships and values to
          identify appropriate criteria.

A weight of evidence approach with multiple attributes that combine one or more of the development
approaches will generally produce criteria of greater scientific validity.

The purpose of this document is to provide guidance on developing numeric nutrient criteria in a scientifically
valid manner, and is not intended to address the multiple, complex issues surrounding implementation of
water quality criteria and standards. Implementation will be addressed in a different process and additional
implementation assistance will also be provided through other technical assistance projects provided by
EPA. For issues specific to constructed wetlands, States should refer to http://www.epa.gov/owow/wetlands/
watersheds/c wetlands. html.
                                         Nutrient Criteria-Wetlands                                        xi

xii                                       Nutrient Criteria-Wetlands



The purpose of this document is to provide technical guidance to assist States in assessing the nutrient status
of their wetlands by considering water, vegetation, and soil conditions, and to provide technical assistance for
developing regionally-based, scientifically defensible, numeric nutrient criteria for wetlands. In this document,
the term "wetlands" or "wetland systems" refers to wetlands that are considered as "waters of the United
States." However, States may, at their discretion, use this document to develop water quality criteria and
standards for wetlands that are considered waters of the State.

EPA's development of recommended nutrient criteria is part of an initiative by the U.S. Environmental
Protection Agency (USEPA) to address the problem  of cultural eutrophication. In 1998, the EPA published
a report entitled, National Strategy for the Development of Regional Nutrient Criteria (USEPA 1998a). The
report outlines a framework for EPA's development of waterbody-specific technical guidance that can be used
to assess nutrient status and develop region-specific  numeric nutrient criteria. This document is the technical
guidance for developing numeric nutrient criteria for wetlands. Approaches to nutrient criteria development are
similar for freshwater and tidal wetlands, however, this document has a freshwater emphasis. EPA recognizes
that wetlands are different from the other types of waters of the U.S. in that they frequently do not have
standing or flowing water, and the soils and vegetation components are more dominant in these systems than
in the other waterbody types (lakes, streams, estuaries).  Additional, more specific information on sampling
wetlands is available at: http://www.epa.gov/waterscience/criteria/nutrient/guidance/index.html.


Cultural eutrophication (human-caused inputs of excess nutrients in waterbodies) is one of the primary causal
factors that impair surface waters in the U.S. (USEPA  1998a). Both point and nonpoint sources of nutrients
contribute to impairment of water quality. Point source discharges of nutrients are relatively constant and are
controlled by the National Pollutant Discharge Elimination System (NPDES) permitting program. Nonpoint
source pollutant inputs have increased in recent decades, resulting in degraded water quality in many
aquatic systems. Nonpoint sources of nutrients are most commonly intermittent and are usually linked to
runoff, atmospheric deposition, seasonal agricultural activity, and other irregularly occurring events such as
silvicultural activities. Control of nonpoint source pollutants typically focuses on land management activities
and regulation of pollutants released to the atmosphere (Kronvang et al. 2005; Howarth et al. 2002; Carpenter
et al. 1998).

The term eutrophication was coined in reference to lake systems. The use of the term for wetlands can be
problematic due to the confounding nature of hydrodynamics, light, and the differences in the responses of
algae and vegetation. Eutrophication in this document  refers to human-caused inputs of excess nutrients and
is not intended to indicate the same scale or responses  to eutrophication found in lake systems and codified in
the trophic state index for lakes (Carlson  1977). This manual is intended to provide guidance for identifying
deviance from natural conditions with respect to cultural eutrophication in wetland systems. Hydrologic
alteration and pollutants other than excess nutrients may amplify or reduce the effects of nutrient pollution,
making specific responses to nutrient pollution difficult to quantify. EPA recognizes these issues, and presents
recommendations for analyzing wetland systems with  respect to nutrient condition for development of nutrient
criteria in spite of these confounding factors.

Cultural eutrophication is not new; however, traditional efforts at nutrient control have been only moderately
successful. Specifically,  efforts to control nutrients in waterbodies that have multiple nutrient sources (point
and nonpoint sources) have been less effective  in providing satisfactory, timely remedies for enrichment-
                                        Nutrient Criteria-Wetlands                                      1-1

    related problems (Azzellino et al. 2006; Merseburger et al. 2005; Carpenter et al. 1998). Development and
    adoption of numeric nutrient criteria into water quality standards aid State nutrient pollution control efforts
    by providing clear numeric goals for nutrient concentrations. Furthermore, numeric nutrient criteria provide
    specific water quality goals that will assist researchers in designing improved best management practices.


    States are responsible for setting water quality standards to protect the physical, biological, and chemical
    integrity of their waters.  "Water quality standards (WQS) are provisions of State or Federal law which consist
    of a designated use or uses and water quality criteria for such waters to protect such uses.2 Water quality
    standards are to protect public health or welfare,  enhance the quality of the water, and serve the purposes of
    the Act (40 CFR 131.2 and 131.3(i))" (USEPA 1994). A water quality standard defines the goals for a wetland
    by: designating its specific uses, setting criteria to protect those uses, and, establishing an antidegradation
    policy to protect existing water quality.

    Designated uses are a State's concise statements  of its goals and expectations for each of the individual surface
    waters under its jurisdiction. With designated uses, States can work with their publics to identify a collective
    goal for their waters that they intend to strive for as they manage water quality. EPA encourages States to
    evaluate the attainability of these goals and expectations to ensure they have designated the appropriate uses.
    Generally, the effectiveness of designated uses in guiding water quality management programs is greater if

            • Identify specific expectations based on as much data as possible to reduce ambiguity;
            • Recognize and accommodate inherent natural differences among surface water types; and
            • Acknowledge certain human caused conditions that limit the potential to support uses.

    Designated uses may involve a spectrum of expectations depending on the type of wetland and associated
    hydropatterns, where the wetland is situated with respect to natural landscape features and human activity,
    and the historical and anticipated future functions that the  wetland provides.   Criteria to protect specific uses,
    in turn, should reflect these differing expectations where appropriate. The information used in developing the
    technical approaches in this document was drawn from references about studies of wetlands in a wide range
    of conditions, but not wetlands with a high degree of modification (e.g., wetlands that are considered "prior
    converted cropland" or artificial wetlands specifically engineered to protect or improve downstream water

    Water quality criteria may be expressed as numeric values or narrative statements. As of this writing, most
    of the Nation's waterbodies do not have numeric  nutrient criteria, but instead rely on narrative criteria that
    describe the desired condition. Narrative criteria are descriptions of conditions necessary for a water to attain
    the designated uses. An example of a narrative criterion for nutrients is shown below:
       In no case shall nutrient concentrations of a body of water be altered so as to cause an imbalance in natural
       populations of aquatic flora or fauna.
    2EPA published guidance on water quality standards for wetlands in 1990 (USEPA 1990c). Examples of different state approaches for
    standards can be found at: http://www.epa.gov/owow/wetlands/initiatives/.
1-2                                      Nutrient Criteria-Wetlands

Numeric criteria, on the other hand, are values assigned to measurable components of water quality to protect
designated uses, such as the concentration of a specific constituent that is present in the water column. An
example of a numeric criterion for specific waters is shown below:
    (4) Phosphorus Criterion.
    (a) The numeric phosphorus criterion for Class III waters shall be a long-term geometric mean of 10
    ppb, but shall not be lower than the natural conditions of the Class II waters, and shall take into account
    spatial and temporal variability. Achievement of the criterion shall be determined by the methods in this
    subsection. Exceedences of the provisions of the subsection shall not be considered deviations from the
    criterion if they are attributable to the full range of natural spatial and temporal variability, statistical
    variability inherent in sampling and testing procedures, or higher natural background conditions.
In addition to narrative and numeric criteria, some States use numeric translator mechanisms—mechanisms
that translate narrative (qualitative) standards into numeric (quantitative) values for use in evaluating water
quality data. Translator mechanisms may be useful internally by the State agency for water assessment and
management and serve as an intermediate step between numeric and narrative criteria.

Numeric criteria provide distinct interpretations of acceptable and unacceptable conditions, form the
foundation for measurement of environmental quality, and reduce ambiguity for management and enforcement
decisions. The lack of numeric nutrient criteria in State water quality standards for most of the Nation's
waterbodies makes it difficult to assess the condition of waters of the U.S. with respect to nutrients, and
thus hampers the water quality manager's ability to protect designated uses and improve water quality. EPA
encourages States to adopt numeric nutrient criteria into their water quality standards (USEPA 2007b).

Many States have adopted some form of nutrient criteria for surface waters related to maintaining natural
conditions and avoiding nutrient enrichment. Most States with nutrient criteria in their water quality standards
have broad narrative criteria for most waterbodies and may also have site-specific numeric criteria for certain
waters of the State. Established criteria most commonly pertain to phosphorus (P) concentrations in lakes.
Nitrogen (N) criteria, where they have been established,  are usually protective of human health effects or relate
to toxic  effects of ammonia and nitrates. In general, levels of nitrate (10 ppm [mg/L] for drinking water) and
ammonia high enough to be problematic for human health or toxic to aquatic life (1.24 mg N/L at pH = 8 and
25°C) will also cause problems of enhanced algal growth (USEPA 1986).

Numeric nutrient criteria can provide a variety of benefits and may be used in conjunction with State
and Federal biological assessments, Nonpoint Source Programs, Watershed Implementation Plans, and
in development of Total Maximum Daily Loads (TMDLs) to improve resource management and support
watershed protection activities at local, State, and national levels. Information obtained from compiling
existing data and conducting new surveys can provide water quality managers and the public a better
perspective on the condition of State waters. The compiled information can be used to most effectively budget
personnel and financial resources for the protection and restoration of State waters. In a similar manner,
data collected in the criteria development and implementation process can be compared before, during, and
after specific management actions. Analyses of these data can determine the response of the wetland and the
effectiveness of management endeavors.


Water quality can be affected when watersheds are modified by alterations in vegetation, sediment transport,
fertilizer use, industrialization, urbanization, or conversion of native forests and grasslands to agriculture and
silviculture (Turner and Rabalais 1991; Vitousek et al.  1997; Carpenter et al. 1998). Cultural eutrophication,
one of the primary factors causing impairment of U.S. surface waters (USEPA 1998a), results from point and
nonpoint sources of nutrient pollution. Nonpoint source pollutant inputs have increased in recent decades
                                        Nutrient Criteria-Wetlands                                      1-3

    and have degraded water quality in many aquatic systems (Carpenter et al. 1998). Control of nonpoint source
    pollutants focuses on land management activities and regulation of pollutants released to the atmosphere
    (Carpenter et al. 1998).

    Nutrient enrichment frequently ranks as one of the top causes of water resource impairment. EPA reported to
    Congress that of the waterbodies surveyed and reported impaired 20 percent of rivers and 50 percent of lakes
    were listed with nutrients as the primary cause of impairment (USEPA 2000c). Few States currently include
    wetland monitoring in their routine water quality monitoring programs (only eleven States reported attainment
    of designated uses for wetlands in the National Water Quality Inventory 1998 Report to Congress (USEPA
    1998b) and only three States used monitoring data as a basis for determining attainment of water quality
    standards for wetlands); thus, the extent of nutrient enrichment and impairment of wetland systems is largely
    undocumented. Increased wetland monitoring by States will help define the extent of nutrient enrichment
    problems in wetland systems.

    The best documented case of cultural eutrophication in wetlands is the Everglades ecosystem. The Everglades
    ecosystem is a wetland mosaic that is composed primarily of oligotrophic freshwater marsh. Historically,
    the greater Everglades ecosystem included vast acreage of freshwater marsh, small stands of custard apple
    and some cattail south of Lake Okeechobee, and Big Cypress Swamp, which eventually drains into Florida
    Bay. Lake Okeechobee was diked to reduce flooding. The area directly south of Lake Okeechobee was then
    converted into agricultural lands for cattle grazing and row crop production. The cultivation and use of
    commercial fertilizers in the area now known as the "Everglades Agricultural Area" have resulted in release
    of nutrient-rich waters into the Everglades for more than thirty years. The effects of the nutrient-rich water,
    combined with coastal development and channeling to supply water to communities on the southern Florida
    coast, have significantly increased soil and water column phosphorus levels in naturally oligotrophic areas. In
    particular, nutrient enrichment of the freshwater marsh has resulted in an imbalance in the native vegetation.
    Cattail is now encroaching in areas that were historically primarily sawgrass; calcareous algal mats are
    being replaced by non-calcareous algae,  changing the balance of native flora that is needed to support vast
    quantities of wildlife. Nutrient enriched water is also reaching Florida Bay, suffocating the native turtle grass
    as periphyton covers the blades (Davis and Ogden 1994; Everglades Interim Report 1999 2003; Everglades
    Consolidated Report 2003). Current efforts to restore the Everglades are focusing on nutrient reduction and
    better hydrologic management (Everglades Consolidated Report 2003).

    Monitoring to establish trends in nutrient levels  and associated changes in biology has been infrequent for
    most wetland types as compared to studies in the Everglades or examination of other surface waters such as
    lakes. Noe et al., (2001) have argued that phosphorus biogeochemistry and the extreme oligotrophy observed
    in the Everglades in the absence of anthropogenic inputs represents a unique case. Effects of cultural
    eutrophication, however, have been documented in a range of different wetland types. Existing studies are
    available to document potential impacts of anthropogenic nutrient additions to a wide variety of wetland types,
    including bogs, fens,  Great Lakes coastal emergent marshes, and cypress swamps. The evidence of nutrient
    effects in wetlands ranges from controlled experimental manipulations, to trend  or empirical gradient analysis,
    to anecdotal observations. Consequences of cultural eutrophication have been observed at both community
    and ecosystem-level scales (Table 1). Changes in wetland vegetation composition resulting from cultural
    eutrophication of these systems have been demonstrated in bogs (Kadlec and Bevis 1990),  fens (Guesewell et
    al. 1998; Bollens and Ramseier 2001, Pauli et al. 2002), meadows (Finlayson et al. 1986), marshes (Bedford et
    al. 1999) and cypress domes (Ewel 1976). Specific effects on higher trophic levels in marshes seem to depend
    on trophic structure (e.g., presence/absence of minnows, benthivores, and/or piscivores, Jude and Pappas 1992;
    Angeler et al. 2003) and timing/frequency of nutrient additions (pulse vs. press; Gabor et al. 1994; Murkin et
    al. 1994; Hann and Goldsborough 1997; Sandilands et al. 2000; Hann et al. 2001; Zrum and Hann 2002).

    The cycling of nitrogen (N) and phosphorus (P)  in aquatic systems should be considered when managing
    nutrient enrichment. The hydroperiod of wetland systems significantly affects  nutrient transformations,
1-4                                     Nutrient Criteria-Wetlands

availability, transport, and loss of gaseous forms to the atmosphere (Mitsch and Gosselink 2000). Nutrients
can be re-introduced into a wetland from the sediment, or by microbial transformation, potentially resulting in
a long recovery period even after pollutant sources have been reduced. In open wetland systems, nutrients may
also be rapidly transported downstream, uncoupling the effects of nutrient inputs from the nutrient source,
and further complicating nutrient source control (Mitsch and Gosselink 2000; Wetzel 2001). Recognizing
relationships between nutrient input and wetland response is the first step in mitigating the effects of cultural
eutrophication. When relationships are established, nutrient criteria can be developed to manage nutrient
pollution and protect wetlands from eutrophication.
Loss of submerged aquatic plants that have high light
compensation points
Shifts in vascular plant species composition due to shifts in
competitive advantage
Increases in above-ground production
Decreases in local or regional biodiversity
Increased competitive advantage of aggressive/invasive species
(e.g., Typha glauca, T. latifolia and Phalaris arundinacea)
Loss of nutrient retention capacity (e.g., carbon and nitrogen
storage, changes in plant litter decomposition)
Major structural shifts between "clear water" macrophyte
dominated systems to turbid phytoplankton dominated systems
or metaphyton-dominated systems with reduced macrophyte
Shifts in macroinvertebrate composition along a cultural
eutrophication gradient
Phillips et al. 1978
Stephenson et al. 1980
Galatowitsch and van der Valk 1996
Wentz 1976
Verhoeven et al. 1988
Ehrenfeld and Schneider 1 993
Gaudet and Keddy 1995
Koerselman and Verhoeven 1995
Barko 1983
Bayleyetal. 1985
Barko and Smart 1986
Vermeer 1986
Mudroch and Capobianco 1979
Guntenspergen et al. 1980
Lougheed et al. 2001
Balla and Davis 1995
VanGroenendael et al. 1993
Bedford etal. 1999
Woo and Zedler 2002
Svengsouk and Mitsch 200 1
Green and Galatowitsch 2002
Maurer and Zedler 2002
Nichols 1983
Davis and van der Valk 1983
Rybczyketal. 1996
McDougal et al. 1997
Angeler et al. 2003
Chessman et al. 2002
        Table 1: Observed consequences of cultural eutrophication in freshwater wetlands.3
3Similar impacts in tidal and estuarine wetlands have been documented, but are not included in this table.
                                          Nutrient Criteria-Wetlands


    The National Strategy for the Development of Regional Nutrient Criteria (USEPA 1998a) describes the
    principal elements of numeric nutrient criteria development. This document can be downloaded in PDF format
    at the Web site: http://www.epa.gov/waterscience/criteria/nutrient/strategy. The Strategy recognizes that a
    prescriptive, one-size-fits-all approach is not appropriate due to regional differences that exist and the scientific
    community's current technical understanding of the relationship between nutrients, algal and macrophyte
    growth, and other factors (e.g., flow, light, substrata). The approach chosen for criteria development therefore
    may be tailored to meet the specific needs of each State. The EPA Strategy envisions a process by which State
    waters are initially monitored, reference conditions are established, individual waterbodies are compared to
    known reference waterbodies, and appropriate management measures are implemented. These measurements
    can be used to document change and monitor the progress of nutrient reduction activities and protection of
    water quality.

    The National Nutrient Program represents an effort and approach to criteria development that, in conjunction
    with efforts made by State water quality managers, will ultimately result in a heightened understanding of
    nutrient-response relationships. As the proposed process is put into use to set criteria, program success will
    be gauged over time through evaluation of management and monitoring efforts. A more comprehensive
    knowledge-base pertaining to nutrient, and vegetation and/or algal relationships will be expanded as new
    information is gained and obstacles overcome, justifying potential refinements to the criteria development

    The overarching goal of developing and adopting nutrient criteria is to protect and maintain the quality of
    our national waters. Protecting and maintaining water quality may include restoration of impaired systems,
    conservation of high quality waters, and protection of systems at high risk for future  impairment. The specific
    goals of a State water quality program may be defined differently based on the needs of each State,  but should,
    at a minimum, be established to protect the designated uses for the waterbodies within State lands.  In addition,
    as numeric nutrient criteria are developed for the nation's waters, States should revisit their goals for water
    quality and revise their water quality standards  as needed.


    As set out in Figure 1.1, the process of developing numeric nutrient criteria begins with defining the goals of
    criteria development and water quality standards adoption. Those goals are pertinent to the classification of
    systems, the development of a monitoring program, and the application of numeric nutrient criteria to permit
    limits and water quality protection. These goals therefore should be determined with the intent of revising and
    adapting them as new information is obtained and the paths to achieving those goals  are clarified. Defining
    the goals for criteria development is the first step in the process. The summaries below describe  each chapter
    in this document. The document is written to provide recommendations for a stepwise procedure for criteria
    development. Some chapters contain information that is not needed by some readers; the descriptions below
    should serve as a guide to the most relevant information for each reader.

    Chapter Two describes many of the functions of wetland systems and their role in the landscape with respect
    to nutrients. This chapter is intended to familiarize the reader with some basic scientific information about
    wetlands that will provide a better understanding of how nutrients move within a wetland and the importance
    of wetland systems in the landscape.

    Chapter Three discusses wetland classification and presents the reader with options for classifying wetlands
    based on system characteristics. This chapter introduces the scientific rationale for classifying wetlands,
    reviews some common classification schemes, and discusses their role in establishing nutrient criteria for
1-6                                      Nutrient Criteria-Wetlands

 wetlands. The classification of these systems is important to identifying their nutrient status and their
condition in relation to similar wetlands.

Chapter Four provides technical guidance on designing effective sampling programs for State wetland water
quality monitoring programs. Most States should begin wetland monitoring programs to collect water quality
and biological data in order to develop nutrient criteria protective of wetland systems. The best monitoring
programs are designed to assess wetland condition with statistical rigor and maximize effective use of available
resources. The sampling protocol selected, therefore, should be determined based on the goals of the monitoring
program, and the resources available.

Chapter Five gives an overview of candidate variables that could be used to establish nutrient criteria for
wetlands. Primary variables are expected to be most broadly useful in characterizing wetland conditions with
respect to nutrients, and include nutrient loading rates, soil nutrient concentrations, and nutrient content of
wetland vegetation. Supporting variables provide information useful for normalizing causal and response
variables. The candidate variables suggested here are not the only parameters that can be used to determine
wetland nutrient condition, but rather identify those variables that are thought to be most likely to identify the
current nutrient condition and of the greatest utility in determining a change in nutrient status.

A database of relevant water quality information can be an invaluable tool to States as they develop nutrient
criteria. If little or no data are available for most regions or parameters, it may be necessary for States to create
a database of newly gathered data. Chapter Six provides the basic information on how to develop a database of
nutrient information for wetlands, and supplies links to ongoing database development efforts at the State and
national levels.

The purpose of Chapter Seven is to  explore methods for  analyzing data that can be used to develop nutrient
criteria. The quality of the analysis and interpretation of data generally determines whether the criteria will
be scientifically defensible and effective. This chapter describes recommended approaches to data analysis for
developing numeric nutrient criteria for wetlands. Included are techniques to evaluate metrics, to examine or
compare distributions of nutrient exposure or response variables, and to examine nutrient exposure-response

Chapter Eight describes the details of establishing scientifically  defensible criteria in wetlands. Several
approaches are presented that water quality managers can use to derive numeric criteria for wetland systems
in their State waters. They include: (1) the use of the reference conditions concept to characterize natural or
minimally impaired wetland systems with respect to causal and response variables; (2) applying predictive
relationships to select nutrient concentrations that will protect wetland function; and, (3) developing criteria
from established nutrient exposure-response relationships (as in the peer-reviewed, published literature). This
chapter provides recommendations regarding how to determine  the appropriate numeric criterion based on the
data collected and analyzed.

The appendices include a glossary of terms and acronyms and case study examples of wetland nutrient
enrichment and management.
                                         Nutrient Criteria-Wetlands                                       1-7

         Figure 1.: Flowchart identifying the steps of the recommended process to develop wetland nutrient
Nutrient Criteria-Wetlands



Wetlands exist at the interface between terrestrial and aquatic environments. They serve as sources, sinks,
and transformers of materials. (Figure 2.1) Wetlands serve as sites for transformation of nutrients such as
nitrogen (N) and phosphorus (P). Dissolved inorganic forms of N and P are assimilated by microorganisms
and vegetation and incorporated into organic compounds. Nitrate in surface- and ground-water is reduced
to gaseous forms of N (NO, N2O, N2) by microorganisms, a process known as denitrification, and returned
to the atmosphere. Phosphorus undergoes a variety of chemical reactions with iron (Fe), aluminum (Al), and
calcium (Ca) that depend on the pH of the soil, availability of sorption sites, redox potential, and other factors.
These biogeochemical reactions are important in evaluating the nutrient condition (oligotrophic, mesotrophic,
eutrophic) of the wetland and  its susceptibility to nutrient enrichment.

Wetlands also generally are sinks for sediment, and wetlands that are connected to adjacent aquatic ecosystems
(e.g., rivers, estuaries) may trap more sediment as compared to wetlands that lack such connectivity (Fryirs
et al. 2007; Mitsch and Gosselink 2000; Dunne and Leopold 1978). Wetlands also may be sources of organic
carbon (C) (Bouchard 2007; Raymond and Bauer 2001) and nitrogen (N) (Mitsch and Gosselink 2000;
Mulholland and Kuenzler 1979) to aquatic ecosystems. Production of plant biomass (leaves, wood, and roots)
from riparian, alluvial, and floodplain forests and from fringe wetlands such as tidal marshes and mangroves
provide organic matter to support heterotrophic foodwebs of streams, rivers, estuaries, and nearshore  waters
(Mitsch and Gosselink 2000; Day et al. 1989).
                                               Atmospheric Inputs
                                                   Atmospheric Outputs I
                                                                   .,  •     . «•
             Surface Water Inflows
       NI Ground Water Inflows
                                                               Ground Water Outflows
                                            Surface Water Outflows
        Figure 2.1: Schematic of nutrient transfer among potential system sources and sinks.
                                      Nutrient Criteria-Wetlands


    Wetlands are distinguished by three primary components: hydrology, soils, and vegetation. Wetland hydrology
    is the driving force that determines soil development, the assemblage of plants and animals that inhabit the
    site, and the type and intensity of biochemical processes. Wetland soils may be either organic or mineral, but
    share the characteristic that they are saturated or flooded at least some of the time during the growing season.
    Wetland vegetation consists of many species of algae, rooted plants that may be herbaceous and emergent, such
    as cattail (Typha sp.) and arrowhead (Saggitaria sp.), or submergent, such as pondweeds (Potamogeton sp.),
    or may be woody such as bald cypress (Taxodium distichum) and tupelo (Nyssa aquatica). Depending on the
    duration, depth,  and frequency of inundation or saturation, wetland plants may be either obligate (i.e., species
    found almost exclusively in wetlands) or facultative (i.e., species found in wetlands but which also may be
    found in upland habitats). The discussion that follows provides an overview of wetland hydrology, soils, and
    vegetation, as well as aspects of biogeochemical cycling in these systems.

    Hydrology is characterized by water source, hydroperiod (depth, duration, and frequency of inundation or
    soil saturation), and hydrodynamics (direction and velocity of water movement). The hydrology of wetlands
    differs from that of terrestrial ecosystems in that wetlands are inundated or saturated long enough during
    the growing season to produce soils that are at least periodically deficient in oxygen. Wetlands differ from
    other aquatic ecosystems by their shallow depth of inundation that enables rooted vegetation to become
    established, in contrast to deep water aquatic ecosystems, where the depth and duration of inundation can be
    too great to support emergent vegetation. Anaerobic soils promote colonization by vegetation adapted to low
    concentrations of oxygen in the soil.

    Wetlands primarily receive water from three sources: precipitation, surface flow, and groundwater (Figure 2.2).
    The relative proportion of these hydrologic inputs influences the plant communities that develop, the types
    of soils that form, and the predominant biogeochemical processes. Wetlands that receive mostly precipitation
    tend to be "closed" systems with little exchange of materials with adjacent terrestrial or aquatic ecosystems.
    Examples of precipitation-driven wetlands include "ombrotrophic" bogs and depressional wetlands  such as
    cypress domes and vernal pools. Wetlands that receive water mostly from surface flow tend to be "open"
    systems with large exchanges of water and materials between the wetland and adjacent non-wetland
    ecosystems. Examples include floodplain forests and fringe wetlands such as lakeshore marshes, tidal marshes,
    and mangroves.  Wetlands that receive primarily groundwater inputs tend to have more stable hydroperiods
    than precipitation- and surface water-driven wetlands, and, depending on the underlying bedrock or parent
    material, high concentrations of dissolved inorganic constituents such as calcium (Ca) and magnesium (Mg).
    Fen wetlands and seeps are examples of groundwater-fed wetlands.
                               GROUND WATER  ^^^^^^^^^^^   PRECIPITATION

                                             33%        67%        100%
                                            	 SURFACE FLOW 	te-
            Figure 2.2: Relationship between water source and wetland vegetation. Modified from Brinson (1993)
2-2                                      Nutrient Criteria-Wetlands

Hydroperiod is highly variable depending on the type of wetland. Some wetlands that receive most of their
water from precipitation (e.g., vernal pools) have very short duration hydroperiods. Wetlands that receive
most of their water from surface flooding (e.g., floodplain swamps) often are flooded longer and to a greater
depth than precipitation-driven wetlands. Fringe wetlands such as tidal marshes and mangroves are frequently
flooded (up to twice daily) by astronomical tides but the duration of inundation is relatively short. In
groundwater-fed wetlands, hydroperiod is more stable and water levels are relatively constant as compared to
precipitation- and surface water-driven wetlands, because groundwater provides a less variable input of water
throughout the year.

Hydrodynamics is especially important in the exchange of materials between wetlands and adjacent
terrestrial and aquatic ecosystems. In fact, the role of wetlands as sources, sinks, and transformers of material
depends, in large part, on hydrodynamics. For example, many wetlands are characterized by lateral flow of
surface- or ground-water. Flow of water can be unidirectional or bidirectional. An example of a wetland with
unidirectional flow is a floodplain forest where surface water spills over the river bank, travels through the
floodplain, and re-enters the river channel some distance downstream. In fringe wetlands such as lakeshore
marshes, tidal marshes, and mangroves, flow is bidirectional as wind-driven or astronomical tides transport
water into, then out of the wetland. These wetlands have the ability to intercept sediment and dissolved
inorganic and organic materials from adjacent systems as water passes through them. In precipitation-driven
wetlands, flow may occur more in the vertical direction as rainfall percolates through the wetland soils to
underlying aquifers or nearby streams. Wetlands with lateral surface flow may be important in maintaining
water quality of adjacent aquatic  systems by trapping sediment and other pollutants. Surface flow wetlands
also may be an important source  of organic C to  aquatic ecosystems as detritus, particulate C, and dissolved
organic C are transported out of the wetland into rivers and streams down gradient or to adjacent lakes,
estuaries, and nearshore waters.

Wetland soils, also known as hydric soils, are defined as "soils that formed under conditions of saturation,
flooding, or ponding long enough during the growing season to develop anaerobic conditions in the upper part"
(NRCS 1998). Anaerobic conditions result because the rate of oxygen diffusion through water is approximately
10,000 times less than in air. Wetland soils may be composed mostly of mineral constituents (sand, silt, clay) or
they may contain large amounts of organic matter. Because anaerobic conditions slow or inhibit decomposition
of organic matter, wetland soils typically contain more organic matter than terrestrial soils of the same
region or climatic conditions. Under conditions of near continuous inundation or saturation, organic soils
(histosols) may develop. Histosols are characterized by high organic matter content, 20-30% (12-18% organic
C depending on clay content) with a thickness of at least 40 cm (USDA  1999). Because of their high organic
matter content, histosols possess  physical and chemical properties that are very different from mineral wetland
soils. For example, organic soils generally have lower bulk densities, higher porosity, greater water holding
capacity, lower nutrient availability, and greater cation exchange capacity than many mineral soils.

Mineral wetland soils, in addition to containing greater amounts of sand, silt, and clay than histosols, are
distinguished by changes in soil color that occur when elements such as Fe and manganese (Mn) are reduced
by microorganisms under anaerobic conditions. Reduction of Fe leads to the development of grey or "gleyed"
soil color as oxidized forms of Fe (ferric Fe, Fe3+) are converted to reduced forms (ferrous Fe, Fe2+). In sandy
soils, development of a dark-colored, organic-rich surface layer is used to distinguish hydric  soil from non-
hydric (terrestrial) soil. An organic-rich surface layer, indicative of periodic inundation or saturation, is not
sufficiently thick (<40 cm) to qualify as a histosol, which forms under near-continuous inundation.

Wetland soils serve as sites for many biogeochemical transformations. They also provide long and short
term storage of nutrients for wetland plants. Wetland soils are typically anaerobic within a few millimeters
of the soil-water  interface. Water column oxygen concentrations are often depressed due to the slow rate of
oxygen diffusion through water. However, even when water column oxygen concentrations are supported by
                                        Nutrient Criteria-Wetlands                                      2-3

    advective currents, high rates of oxygen consumption lead to the formation of a very thin oxidized layer at
    the soil-water interface.  Similar oxidized layers can also be found surrounding roots of wetland plants. Many
    wetland plants are known to transport oxygen into the root zone, thus creating aerobic zones in predominantly
    anaerobic soil. The presence of these aerobic (oxidizing) zones within the reducing environment in saturated
    soils allows for the occurrence of oxidative and reductive transformations to occur in close proximity to each
    other. For example, ammonia is oxidized to nitrate within the aerobic zone surrounding plant roots in a process
    called nitrification. Nitrate then readily diffuses into adjacent anaerobic soil, where  it is reduced to molecular
    nitrogen via denitrification or may be reduced to ammonium in certain conditions through dissimilatory
    nitrate reduction (Mitsch and  Gosselink 2000; Ruckauf et al. 2004; Reddy and Delaune 2005). The anaerobic
    environment hosts the transformations of N, P, sulfur (S), Fe, Mn, and C. Most of these transformations are
    microbially mediated. The oxidized soil surface layer also is important to the transport and translocation
    of transformed constituents, providing a barrier to translocation of some reduced constituents. These
    transformations will be discussed in more detail below in Biogeochemical Cycling.

    Wetland plants consist of macrophytes and microphytes. Macrophytes include free-floating, submersed,
    floating-leaved, and rooted emergent plants. Microphytes are algae that may be free floating or attached
    to macrophyte stems and other surfaces. Plants require oxygen to meet respiration demands for growth,
    metabolism, and reproduction. In macrophytes, much (about 50%) of the respiration occurs below ground in
    the roots. Wetland macrophytes, however, live in periodically to continuously-inundated and saturated soils
    and, therefore, use specialized adaptations to grow in anaerobic soils (Cronk and Fennessy 2001). Adaptations
    consist of morphological/anatomical adaptations that result in anoxia avoidance, and metabolic adaptations
    that result in true tolerance to anoxia. Morphological/anatomic adaptations include shallow roots systems,
    aerenchyma, buttressed  trunks, pneumatophores (e.g., black mangrove (Avicennia germinans)), and lenticles
    on the stem. These adaptations facilitate oxygen transport from the shoots to the roots where most respiration
    occurs. Many wetland plants also possess metabolic adaptations, such as anaerobic pathways of respiration,
    that produce non-toxic metabolites  such as malate to mitigate the adverse effects of oxygen deprivation,
    instead of toxic compounds like ethanol (Mendelssohn and Burdick 1988).

    Species best adapted to anaerobic conditions are typically found in areas inundated  for long periods, whereas
    species less tolerant of anaerobic conditions are found in areas where hydroperiod is shorter. For example,
    in southern forested wetlands, areas such as abandoned river channels (oxbows) are  dominated by obligate
    species such as bald cypress (Taxodium distichum) and tupelo gum (Nyssa aquatica) (Wharton et al. 1982).
    Areas inundated less frequently are dominated by hardwoods such as black gum (Nyssa sylvatica), green ash
    (Fraxinus pennsylvanicus), and red maple (Acer rubrum), and the highest, driest wetland areas are dominated
    by facultative species such as  sweet gum (Liquidambar styraciflua) and sycamore (Platanus occidentalis)
    (Wharton et al. 1982). Herbaceous-dominated wetlands also exhibit patterns of zonation controlled by
    hydroperiod (Mitsch and Gosselink 2000).

    In estuarine wetlands such as salt- and brackish-water marshes and mangroves, salinity and sulfides also
    adversely affect growth  and reproduction of vegetation (Webb and Mendelssohn 1996; Mitsch and Gosselink
    2000). Inundation with seawater brings dissolved salts (NaCl) and sulfate. Salt creates an osmotic imbalance in
    vegetation, leading to desiccation of plant tissues. However, many plant species that live in estuarine wetlands
    possess adaptations to deal with salinity (Winchester et al.  1985; Whipple et al. 1981; Zheng et al. 2004). These
    adaptations include salt exclusion at the root surface, salt secreting glands on leaves, schlerophyllous (thick,
    waxy) leaves, low transpiration rates, and other adaptations to reduce uptake of water and associated salt.
    Sulfate carried in by the tides undergoes sulfate reduction in anaerobic soils to produce hydrogen sulfide (H2S)
    that, at high concentrations, is toxic to vegetation. At sub-lethal concentrations, H2S inhibits nutrient uptake
    and impairs plant growth.
2-4                                      Nutrient Criteria-Wetlands


Point Sources
Point source discharges of nutrients to wetlands may come from municipal or industrial discharges, including
stormwater runoff from municipalities or industries, or in some cases from large animal feeding operations.
Nutrients from point source discharges may be controlled through the National Pollutant Discharge
Elimination System (NPDES) permits, most of which are administered by States authorized to issue them. In
general, point source discharges that are not stormwater related are fairly constant with respect to loadings.

Nonpoint Sources
Nonpoint sources of nutrients are commonly discontinuous and can be linked to seasonal agricultural
activity or other irregularly occurring events such as silviculture, non-regulated construction, and storm
events. Nonpoint nutrient pollution from agriculture is most commonly associated with row crop agriculture
and livestock production that tend to be highly associated with rain events and seasonal land use activities.
Nonpoint nutrient pollution from urban and suburban areas is most often associated with climatological events
(rain, snow, and snowmelt), when pollutants are most likely to be transported to aquatic resources.

Urban and agricultural runoff is generally thought to be the largest source of nonpoint source pollution;
however, growing evidence suggests that atmospheric deposition may have a significant influence on nutrient
enrichment, particularly from nitrogen (Jaworski et al. 1997). Gases released through fossil fuel combustion
and agricultural practices are two major sources of atmospheric N that may be deposited in waterbodies
(Carpenter et al. 1998). Nitrogen and nitrogen compounds formed in the atmosphere return to the earth as acid
rain or snow, gas, or dry particles. Atmospheric deposition, like other forms  of pollution, may be determined
at different scales of resolution. More information on national atmospheric deposition can be  found at: http://
www.arl.noaa.gov/research/programs/airmon.html and http://nadp.sws.uiuc.edu/. These national maps may
provide the user with information about regional areas where atmospheric deposition, particularly of nitrogen,
may be of concern. However, these maps are generally low resolution when considered at the local and site-
specific scale and may not reflect areas of high local atmospheric deposition, such as local areas in a downwind
plume from an animal feedlot operation.

Other nonpoint sources of nutrient pollution may include certain silviculture and mining operations; these
activities generally constitute a smaller fraction of the national problem, but may be locally significant nutrient
sources. Control of nonpoint source pollutants focuses on land management activities and regulation of
pollutants released to the atmosphere (Carpenter et al.  1998).

Wetland nutrient inputs mirror wetland hydrologic inputs (e.g., precipitation, surface water, and ground water),
with additional loading associated with atmospheric dry deposition and nitrogen transformation (Figures 2.5
and 2.6). Total atmospheric deposition (wet and dry) may be the dominant input for precipitation-dominated
wetlands, while surface- or ground-water inputs may dominate other wetland systems.

The total annual nutrient load (mg-nutrients/yr) into a wetland is the sum of the dissolved and particulate loads.
The dissolved load (mg-nutrients/s) can be estimated by multiplying the instantaneous inflow (L/s) by the
nutrient concentration (mg-nutrients/L). EPA recommends calculating the annual load by the summation of this
function over the year—greater loads may be found during periods of increased flow and EPA recommends
monitoring during these intervals. Where continuous data are unavailable, average flows and concentrations
may be used if a bias factor (Cohn et al. 1989) is included to account for unmeasured loads during high flows.
                                        Nutrient Criteria-Wetlands                                      2-5

    Particulate loads (mg-nutrients/yr) can be estimated using the product of suspended and bedload inputs (kg-
    sediments/yr) and the mass concentrations (mg-nutrients/kg-sediment).

    Surface-water nutrient inputs are associated with flows from influent streams, as well as diffuse sources from
    overland flow through the littoral zone. Ground-water inputs can also be concentrated at points (e.g., springs),
    or diffuse (such as seeps). The influence of allochthonous sources is likely to be greatest in those zones closest
    to the source.

    Because wetlands generally tend to be low-velocity, depositional environments, they  often sequester sediments
    and their associated nutrients. These sediment inputs generally accumulate at or near the point of entry into the
    wetland, forming deltas or levees near tributaries, or along the shoreline for littoral inputs. Coarser fractions
    (e.g., gravels and sands) tend to settle first; the finer fractions (silts, clays, and organic matter) tend to settle
    further from the inlet point. Particulate input from ground-water sources can usually be neglected, while
    particulate inputs from atmospheric sources may be important if local or regional sources are present.

    Wetland nutrient outputs again mirror hydrologic outputs (e.g., surface- and ground-water), and loads are
    again estimated as the product of the flow and the concentration of nutrients in the flow. While evaporation
    losses from wetlands may be significant, there are no nutrient losses associated with this loss. Instead,  loss
    of nutrients to the atmosphere may occur as  a result of ammonia volatilization, as well as N2O losses from
    incomplete denitrification. Because sediment outputs from wetlands may be minor, nutrient exports by this
    mechanism may not be important.

    Nutrient accumulation in wetlands occurs when nutrient inputs exceed outputs. Net nutrient loads can be
    estimated as the difference between these inputs and outputs. It is important, therefore, to have some estimate
    of net accumulation by  taking the difference between upstream and downstream loads. Sampling ground-water
    nutrient concentrations in wells located upstream and downstream of the wetland can provide some sense of
    net nutrient sequestration, while sampling wetland nutrient inflows and outflows is needed for determining the
    additional sequestration for this pathway.

    Biogeochemical cycling of nutrients in wetlands is governed by physical, chemical, and biological processes in
    the soil and water column. Biogeochemical cycling of nutrients is not unique to wetlands, but the aerobic and
    anaerobic interface generally found in saturated soils of wetlands creates unique conditions that allow both
    aerobic and anaerobic processes to operate simultaneously. The hydrology and geomorphology of wetlands
    (Johnston et al. 2001) influences biogeochemical processes and constituent transport  and transformation within
    the systems (e.g., water-sediment exchange,  plant uptake, and export of organic matter). Interrelationships
    among hydrology, biogeochemistry, and the  response of wetland biota vary among wetland types (Mitsch and
    Gosselink 2000; Reddy and Delaune 2005).

    Biogeochemical processes in the soil and water column are key drivers of several ecosystem functions
    associated with wetland values (e.g., water quality improvement through denitrification, long-term nutrient
    storage in the organic matter) (Figure 2.3). The hub for biogeochemistry is organic matter and its cycling in
    the soil and water column. Nutrients such as N, P, and S are primary components of soil organic matter, and
    cycling of these nutrients is always coupled to C cycling. Many processes occur within the carbon, nitrogen,
    phosphorus, and sulfur (C, N, P, or S) cycles; microbial communities mediate the rate and extent of these
    reactions in soil and the water column.

    Aerobic-anaerobic interfaces are more common in wetlands than in upland landscapes and may occur at the
    soil water interface, in the root zones of aquatic macrophytes, and at surfaces of detrital tissue and benthic
    periphyton mats. The juxtaposition of aerobic and anaerobic zones in wetlands supports a wide range of
    microbial populations and associated metabolic activities, with oxygen reduction occurring in the aerobic
2-6                                      Nutrient Criteria-Wetlands


          Figure 2.3: Schematic showing basic nutrient cycles in soil-water column of a wetland.

interface of the substrate, and reduction of alternate electron acceptors occurring in the anaerobic zone
(D'Angelo and Reddy 1994a or b). Under continuously saturated soil conditions, vertical layering of different
metabolic activities can be present, with oxygen reduction occurring at and just below the soil floodwater
interface. Substantial aerobic decomposition of plant detritus occurs in the water column; however, the supply
of oxygen may be insufficient to meet demands and drive certain microbial groups to utilize alternate electron
acceptors (e.g., nitrate, oxidized forms of iron (Fe) and manganese (Mn), sulfate, and bicarbonate (HCO3)).

Soil drainage adds oxygen to the soil, while other inorganic electron acceptors may be added through hydraulic
loading to the system. Draining wetland soil accelerates organic matter decomposition due to the introduction
of oxygen deeper into the profile. In many wetlands, the influence of NO3 and oxidized forms of Mn and Fe
on organic matter decomposition is minimal. This is because the concentrations of these electron acceptors
are usually low as a result of the fact that they have greater reduction potential than other alternate electron
acceptors, so they generally are depleted rapidly from systems. Long-term sustainable microbial activity is
then supported by electron acceptors of lower reduction potentials (sulfate and HCO3). Methanogenesis is often
viewed as the terminal step in anaerobic decomposition in freshwater wetlands, whereas sulfate reduction is
viewed as the dominant process in coastal wetlands. However, both processes can function simultaneously in
the same ecosystem and compete for available  substrates (Capone and Kiene 1988).

A simple way to characterize wetlands for aerobic and anaerobic zones is to determine the oxidation-reduction
potential or redox potential (Eh) of the soil-water column (Figure 2.4). Redox potential is expressed in units of
millivolts (mV) and is measured using a voltmeter coupled to a platinum electrode and a reference electrode.
Typically, wetland soils with Eh values >300 mV are considered aerobic and typical of drained soil conditions.
while soils with Eh values <300 mV are considered anaerobic and are devoid of molecular oxygen (Figure 2.4).
                                       Nutrient Criteria-Wetlands

              Flooded Soil
                                              Drained Soil
-100      0       100             300              500
       Oxidation-Reduction Potential (mV)
           Figure 2.4: Range of redox potentials in wetland soils (Reddy and Delaune 2007).
    Wetlands, as low-lying areas in the landscape, receive inputs from all hydrologically connected uplands.
    Many wetlands are open systems receiving inputs of carbon (C) and nutrients from upstream portions of the
    watershed that can include agricultural and urban areas.

    Prolonged nutrient loading to wetlands can result in distinct gradients in water and soil. Mass loading and
    hydraulic retention time determine the degree and extent of nutrient enrichment. Continual nutrient loading
    to an oligotrophic wetland can result in a zone of high nutrient availability near the input, and low nutrient
    availability and possibly nutrient limiting conditions further from the input point. This enrichment effect can
    be seen in many freshwater wetlands, most notably in the sub-tropical Everglades where light is abundant and
    temperatures are high (Davis 1991; Reddy et al. 1993; Craft and Richardson 1993 a, b; DeBusk et al. 1994), and
    in some estuarine marshes (Morris and Bradley 1999). Between these two extremes, there can exist a gradient
    in quality and quantity of organic matter, nutrient accumulation, microbial and macrobiotic communities,
    composition, and biogeochemical cycles.

    Compared to terrestrial ecosystems, most wetlands show an accumulation of organic matter, and therefore
    wetlands function as global sinks for carbon. Accumulation of organic C in wetlands is primarily a result of
    the balance of C fixation through photosynthesis and losses through decomposition. Rates of photosynthesis in
    wetlands are typically higher than in other ecosystems, and rates of decomposition are typically lower due to
    anaerobic conditions, hence organic matter tends to accumulate. In addition to maintaining proper functioning
    of wetlands, organic matter storage also plays an important role in regulating other ecosystems and the
    biosphere. For example, organic matter contains substantial quantities of N,  P, and S; therefore, accumulation
    of organic matter in wetlands decreases transport of these nutrients to downstream aquatic systems.
                 Nutrient Criteria-Wetlands

                                                    Plant biomass
 N2 N2O
                                                             Mineralization  .....  +   ....   =—,	
                                                Organic    -^—•—^^-  NH4   Water Column

                                             Organic N •**• Biomass N   Adsorbed NH4+
   ^— N2, N20 (g)
        Figure 2.5: Schematic of the nitrogen cycle in wetlands.

Nitrogen enters wetlands in organic and inorganic forms, with the relative proportion of each depending on
the input source. Organic forms are present in dissolved and particulate fractions, while inorganic N (NH4-N,
NO3-N and NO2-N) is present in dissolved fractions (Figure 2.5) or bound to suspended sediments (NH4-N).
Particulate fractions are removed through settling and burial, while the removal of dissolved forms is regulated
by various biogeochemical reactions functioning in the soil and water column. Relative rates of these processes
are affected by physico-chemical and biological characteristics of plants, algae, and microorganisms.

Nitrogen reactions in wetlands effectively process inorganic N through nitrification and denitrification.
ammonia volatilization, and plant uptake. These processes aid in lowering levels of inorganic N in the water
column. A significant portion of dissolved organic N assimilated by plants is returned to the water column
during breakdown of detrital tissue or soil organic matter, and the majority of this dissolved organic N is
resistant to decomposition. Under these conditions, water leaving wetlands may contain elevated levels of
N in organic form. Exchange of dissolved nitrogen species between soil and water column support several
nitrogen reactions. For example, nitrification in the aerobic soil layer is supported by ammonium flux from
the anaerobic soil layer. Similarly,  denitrification in the anaerobic soil layer is supported by nitrate flux
from the  aerobic soil layer and water column. Relative rates of these reactions will, however, depend on the
environmental conditions present in the soil and water column (Reddy and Delaune 2007).

Phosphorus retention by wetlands is regulated by physical (sedimentation and entrainment), chemical
(precipitation and flocculation), and biological mechanisms (uptake and release by vegetation, periphyton.
and microorganisms). Phosphorus in the influent water is found in soluble and particulate fractions, with both
fractions containing a certain proportion of inorganic and organic forms.  Relative proportions of these pools
depend on the input source. For example, municipal wastewater may contain a large proportion (>75%) as
inorganic P in soluble forms, as compared to effluents from agricultural watersheds where a greater percentage
of P  loading may be in the particulate fraction.
                                        Nutrient Criteria-Wetlands

        Plant biomass P
                                                                                       Water Column
                                                                                          [Fe, Al or
                                                                                        Soil - ANAEROBIC
            Figure 2.6: Schematic of the phosphorous cycle in wetlands.

    Phosphorus forms that enter a wetland are grouped into: (i) dissolved inorganic P (DIP); (ii) dissolved organic
    P (DOP); (iii) particulate inorganic P (PIP); and, (iv) particulate organic P (POP) (Figure 2.6). The particulate
    and soluble organic fractions may be further separated into labile and refractory components. Dissolved
    inorganic P is generally bioavailable, whereas organic and particulate P forms generally must be transformed
    into inorganic forms before becoming bioavailable. Both biotic and abiotic mechanisms regulate relative pool
    sizes and transformations of P compounds within the water column and soil. Alterations in these fractions can
    occur during flow through wetlands and depend on the physical, chemical, and biological characteristics of the
    systems. Thus, both biotic and abiotic processes should be considered when evaluating P retention capacities
    of wetlands. Biotic processes include assimilation by vegetation, plankton, periphyton, and microorganisms.
    Abiotic processes include sedimentation, adsorption by soils, precipitation, and exchange processes between
    soil and the overlying water column (Reddy et al. 1999, 2005; Reddy and Delaune 2007). The processes
    affecting phosphorus exchange at the soil/sediment water interface include:  (i) diffusion and advection due
    to wind-driven currents; (ii) diffusion and advection due to flow and bioturbation; (iii) processes within the
    water column (mineralization, sorption by particulate matter, and biotic uptake and release); (iv) diagenetic
    processes (mineralization, sorption, and precipitation dissolution) in bottom sediments; (v) redox conditions
    (O2 content) at the soil/sediment-water interface; and, (vi) phosphorus flux from water column to soil mediated
    by evapotranspiration by vegetation.

    The key biogeochemical services provided by wetlands include nutrient transformation and removal by
    decreasing concentrations of nutrients and other contaminants, and sequestration of carbon and nutrients into
    stable pools (Kadlec and Knight 1996). The biogeochemical processes regulating water quality improvement
    are well established, and are made use of in treatment wetlands. Increased nutrient loading to oligotrophic
    wetlands results in increased primary productivity and nutrient enrichment. This resulting eutrophication can
    have both positive and negative impacts to the environment. Higher rates of primary productivity increase
    rates of organic matter accumulation, thus increasing carbon sequestration. However, eutrophication may lead
    to increased periodic and episodic export of DIP (Kadlec and Knight 1996; Reddy et al. 1995, 1996, 2005;
    Reddy and Delaune 2007).
      Nutrient Criteria-Wetlands



Developing individual, site-specific nutrient criteria is not practical for every wetland. Instead, criteria for
groups of similar wetlands in a region are needed. To this end, a means of grouping or classifying wetlands
is required. This chapter introduces the scientific rationale for classifying wetlands, reviews some common
classification schemes, and discusses their implications for establishing nutrient criteria for wetlands. Use
of a common scheme across State boundaries should facilitate collaborative efforts in describing reference
condition for biota or water quality and in developing assessment methods, indices of biotic integrity (IBI)
(USEPA 1993b, http://www.epa.gov/emap/remap/index.html), nutrient-response relationships, and nutrient
criteria for wetlands. This chapter describes a series of national classification systems that could be used to
provide a common framework for development of nutrient criteria for wetlands, and suggests ways in which
these classification schemes could be combined in a hierarchical fashion. Many existing classification schemes
may be relevant and should be considered for use or modification, even if they were not originally derived
for wetland nutrient criteria because: (1) they incorporate key factors that control nutrient inputs and cycling;
(2) they have already been mapped; and,  (3) they have been incorporated into sampling, assessment, and
management strategies for wetland biology or for other surface water types, thus facilitating integration of
monitoring strategies. Adoption of any classification scheme should be an iterative process, whereby initial
results of biological  or water quality sampling are used to test for actual differences in reference condition
for nutrients or nutrient-response relationships among proposed wetland classes. Wetland classes that behave
similarly can be combined, and apparent outliers in distributions of nutrient concentrations from reference
sites or in nutrient-response relationships can be examined for additional sources of variability that may need
to  be considered. In addition, new classification schemes can be derived empirically through many multivariate
statistical methods designed to determine factors that can discriminate among wetlands based on nutrient
levels or nutrient-response relationships.

The overall goal of classification is to reduce variability within classes due to differences in natural condition
related to factors such as geology, hydrology, and climate. This will minimize the number of classes for
which reference conditions must be defined. For example, we would expect different conditions for water
quality or biological community composition for wetland classes in organic soils (histosols), compared to
wetlands in mineral soils. In assessing impacts to wetlands, comparing a wetland from within the same class
would increase the precision of assessments, enable more sensitive detection of change, and reduce errors in
characterizing the status of wetland condition.

Reference conditions "describe the characteristics of waterbody segments least impaired by human activities
and are used to define attainable biological  or habitat conditions" (USEPA 1990; Stoddard et al. 2006). At least
two general approaches have been defined to establish reference condition—the site-specific and the regional
(U.S. EPA 1990b, http://www.epa.gov/bioindicators/). The current approach to developing water quality
criteria for nutrients also emphasizes the identification of expected ranges of nutrients by waterbody type
and ecoregion for the least-impaired reference conditions (U.S. EPA 1998; http://www.epa.gov/waterscience/

Although different concepts of reference condition have been used in other programs (e.g., for evaluation of
wetland mitigation projects (Smith et al.  1995; http://el.erdc.usace.army.mil/wetlands/pdfs/wrpde9.pdf)),  for
the purposes of this  document, the term "reference condition" refers to wetlands that are minimally or least
impacted by human activities. Most, if not all, wetlands in the U.S. are affected to some extent by human
activities such as acid precipitation, global climate change, or other atmospheric deposition of nitrogen and
mercury, and changes in historic fire regime. "Minimally impacted" is therefore operationally defined by
                                         Nutrient Criteria-Wetlands                                       3-1

    choosing sites with fewer stressors or fewer overall impacts as described by indicators of stressors, such as
    land-use or human activities within the watershed or buffer area surrounding a wetland and source inputs.
    Identifying reference wetlands in areas of high local or regional atmospheric deposition of nitrogen should also
    be carefully considered because indicators such as local land use activities may not be sufficient to indicate
    nutrient enrichment from dry or wet air deposition.


    There are two different approaches for classification of aquatic resources. One is geographically-based,
    and the other is independent of geography but relies on environmental characteristics that determine
    aquatic ecosystem status and vulnerability at the region-, watershed-, or ecosystem-scale (Detenbeck et al.
    2000). Ecoregions (including "nutrient ecoregions") and Ecological Units represent geographically-based
    classification schemes that have been developed and applied nation-wide (Omernik 1987; Keys et al. 1995).
    The goal of geographically-based classification schemes is to reduce variability in reference condition based
    on spatial co-variance in climate and geology, along with topography, vegetation, hydrology, and soils.
    Geographically-independent or environmentally-based classification schemes include those derived using
    watershed characteristics such as land-use and/or land-cover (Detenbeck et al. 2000), hydro geomorphology
    (Brinson 1993), vegetation type (Grossman et al. 1998), or some combination of these (Cowardin et al. 1979).
    Both geographically- and environmentally-based schemes have been developed for wetland classification.
    These approaches can be applied individually or combined within a hierarchical framework (Detenbeck et al.

    Regional classification systems were first developed specifically for the United States by land management
    agencies. The U.S. Department of Agriculture (USDA) has described a hierarchical system of Land Resource
    Regions and Major Land Resource Areas based mainly on soil characteristics for agricultural management
    (USDA SCS 1981). Ecoregions were then refined for USDA and the U.S. Forest Service based on a hierarchical
    system in which each of several environmental variables such as climate, landform, and potential natural
    vegetation were applied to define different levels of classification (Bailey 1976). Subsequently, Omernik and
    colleagues developed a hierarchical, nationwide ecoregion system to classify streams using environmental
    features they expected would influence aquatic resources, as opposed to terrestrial resources (Hughes and
    Omernik 1981; Omernik et al. 1982). The latter was based on an overlay of "component maps" for land use,
    potential natural vegetation, land-surface form,  and soils, along with a subjective evaluation of the spatial
    congruence of these factors as compared to the hierarchical approach used by Bailey, which relied only on
    natural features (not land-use). Omernik has produced a national map of 84 ecoregions defined at a scale
    of 1:7,500,000 (Figure 3.1; Omernik 1987, http://water.usgs.gov/GIS/metadata/usgswrd/XML/ecoregion.
    xml). More detailed, regional maps have been prepared at a scale of 1:2,500,000 in which the most "typical"
    areas within each ecoregion are defined. Cowardin et al., (1979) have suggested an amendment to  Bailey's
    ecoregions to include coastal and estuarine waters (Figure 3.2a). In practice,  Omernik's scheme has been more
    widely used for geographic classification of aquatic resources such as streams, but few examples to verify the
    appropriateness of this grouping to wetland nutrients are available.

    Finally, an attempt has been made to integrate approaches across Federal agencies to produce regional
    boundaries termed Ecological Units (Keys et al.  1995). Information has been combined on climate, landform,
    geomorphology, geology, soils, hydrology, and potential vegetation to produce a nested series of boundaries for
    the eastern U.S. Different combinations of environmental parameters are emphasized at each hierarchical level
    of classification. This scheme was developed to  explain variation in both terrestrial and aquatic  systems, and
    is consistent with a more comprehensive strategy to classify lotic systems down to the level of stream reaches
    (Maxwell et al.  1995). The mapped system for the eastern U.S. includes classification at the following levels:

            domain (n=2) > divisions (n=5) > provinces (n=14) > sections (n=78) > subsections (n=xxx),
3-2                                      Nutrient Criteria-Wetlands

where Sections are roughly half the size of Omernik ecoregions (Figure 3.3). For lotic systems, additional
spatial detail can be added by defining watersheds (at the level of land type associations), subwatersheds (at the
level of land types), valley segments, stream reaches, and, finally, channel units (Maxwell et al. 1995). In reality,
not all watersheds nest neatly within subsections, and may cross-subsection boundaries.
         Figure 3.1: Map of Omernik aquatic ecoregions.
                                                                               5. WEST
                                                                                  BOUNDARIES OF
                                                                                  10 MARINE AND ESTUARINE
         Figure 3.2a:  Map of Bailey ecoregions with coastal and estuarine provinces.  (Cowardin et al. 1979).
                                          Nutrient Criteria-Wetlands

     a Domains, Divisions, Provinces, and Sections used on Bailey's (1976) map and described in detail in Bailey
     (1978). Highland ecoregions are designated M mountain, P plateau, and A altiplano.
     1000 Polar
          1200 Tundra
          1210 Arctic Tundra
          1220 Bering Tundra
               M1210 Brooks Range
          1300 Subarctic
          1310 Yukon Parkland
          1320 Yukon Forest
               M1310 Alaska Range
     2000 Humid Temperate
          2100 Warm Continental
          2110 Laurentian Mixed Forest
          2111 Spruce-Fir Forest
          2112 Northern Hardwoods-Fir Forest
          2113 Northern Hardwoods Forest
          2114 Northern Harwoods-Spruce Forest
               M2110 Columbia Forest
               M2 111 Douglas-fir Forest
               M2112 Cedar-Hemlock-Douglas-fir Forest
     2200 Hot Continental
          2210 Eastern Deciduous  Forest
          2211 Mixed Mesophytic  Forest
          2212 Beech-Maple Forest
          2213 Maple-Basswood Forest + Oak Savanna
          2214 Appalachian Oak Forest
          2215 Oak-Hickory Forest
     2300 Subtropical
          2310 Outer Coastal Plain Forest
          2311 Beech-Sweetgum-Magnolia-Pine-Oak
          2312 Southern Floodplain Forest
          2320 Southeastern Mixed Forest
     2400 Marine
          2410 Willamette-Puget Forest
               M2410 Pacific Forest (in conterminous U.S.)
               M2411 Sitka Spruce-Cedar-Hemlock Forest
               M2412 Redwood Forest
               M2413 Cedar-Hemlock-Douglas-fir Forest
               M2414 California  Mixed Evergreen Forest
               M2415 Silver fir-Douglas-fir Forest
               M2410 Pacific Forest (in Alaska)
     2500 Prairie
          2510 Prairie Parkland
          2511 Oak-Hickory-Bluestem Parkland
          2512 Oak+Bluestem Parkland
          2520 Prairie Brushland
          2521 Mesquite-Buffalo Grass
          2522 Juniper-Oak-Mesquite
          2523 Mesquite-Acacia
          2530 Tall-Grass Prairie
          2531 Bluestem Prairie
          2532 Whestgrass-Bluestem-Needlegrass
          2533 Bluestem-Gamma Prairie
     2600 Mediterranean (Dry-summer Subtropical)
          2610 California Grassland
               M2610 Sierran Forest
               M2620 California  Chaparral
                 3000 Dry
                       3100 Steppe
                       3110 Great Planins-Shortgrass Prairie
                       3111 Gramma-Needlegrass-Wheatgrass
                       3112 Wheatgrass-Needlegrass
                       3113 Grama-Buffalo Grass
                            M3110 Rocky Mountain Forest
                            M3 111 Grand-fir-Douglas-fir Forest
                            M3112 Douglas-fir Forest
                            M3113 Ponderosa Pine-Douglas-fir Forest
                       3120 Palouse Grassland
                            M3120 Upper Gila Mountains Forest
                       3130 Intermountain Sagebrush
                       3131 Sagebrush-Wheatgrass
                       3132 Lahontan Saltbush-Greasewood
                       3133 Great Basin Sagebrush
                       3134 Bonneville Saltbush-Greasewood
                       3135 Ponderosa Shrub Forest
                            P3130 Colorado Plateau
                            P3131 Juniper-Pinyon Woodland + Sagebrush Saltbush
                            P3132 Grama-Galleta Steppe + Juniper-Pinyon
                                  Woodland Mosaic
                       3140 Mexican Highland Shrub Steppe
                           A3140 Wyoming Basin
                           A3141 Wheatgrass-Needlegrass-Sagebrush
                           A3142 Sagebrush-Wheatgrass
                       3200 Desert 3210 Chihuahuan Desert
                       3211 Grama-Tobosa
                       3212 Tarbush-Creosote Bush
                       3220 American Desert
                       3221 Creosote Bush
                       3222 Creosote Bush-Bur Sage
                 4000 Humid Tropical
                       4100 Savanna
                       4110 Everglades
                       4200 Rainforest
                            M4210 Hawaiian Islands
              Figure 3.2b: Legend for Bailey ecoregion map shown in Figure 3.2a.
Nutrient Criteria-Wetlands

        Figure 3.3: Examples of first four hierarchical levels of Ecological Units: domain, division, province,
        and section, from USEPA Environmental Atlas.

Some States have chosen to refine the spatial resolution of Omernik's ecoregional boundaries for management
of aquatic resources (e.g., Region 3  and Florida). For example, the State of Florida has defined subecoregions
for streams based on analysis of macroinvertebrate data from 100 minimally-impacted sites. Efforts are currently
underway to define ecoregions for Florida wetlands based on variables influencing the water budget and plant
community composition (Dougherty et al. 2000; Lane 2000).
                                         Nutrient Criteria-Wetlands

    Wetland habitat types are described very simply but coarsely by Shaw and Fredine (1956, Circular 39), ranging
    from temporarily-flooded systems to ponds. A more refined hierarchical  classification system is available
    based on vegetation associations; for example, the system developed by the Nature Conservancy for terrestrial
    vegetation includes some wetland types (Grossman et al. 1998). Vegetation associations have also been used to
    classify Great Lakes coastal wetlands within coastal geomorphic type (Michigan Natural Features Inventory

    The Cowardin classification system (Cowardin et al. 1979) was developed for the U.S. Fish and Wildlife Service
    (F WS) as a basis for identifying, classifying, and mapping wetlands, special aquatic sites, and deepwater aquatic
    habitats. The Cowardin system combines a number of approaches incorporating landscape position, hydrologic
    regime, and habitat (vegetative) type (http://www.nwi.fws.gov) (Figure 3.4). Wetlands are categorized first by
    landscape position (tidal, riverine, lacustrine, and palustrine), then by cover  type (e.g., open water,  submerged
    aquatic bed, persistent emergent vegetation, shrub wetlands,  and forested wetlands), and then by hydrologic
    regime  (ranging from saturated or temporarily-flooded to permanently flooded). Modifiers  can  be added for
    different salinity or acidity classes, soil type (organic vs. mineral), or disturbance activities  (impoundment,
    beaver activity). Thus, the Cowardin system includes a mixture of geographically-based  factors, proximal
    forcing functions (hydrologic regime, acidity), anthropogenic disturbance regimes, and vegetative outcomes.
    In practice,  the Cowardin  system can be aggregated by combination of hydrogeomorphic  (HGM) type and
    predominant vegetation cover if digital coverages are available (Ernst et al. 1995).
                  C	Iff	*f*	*

^-  LJ   -
3  g   <
5  oo   £
                                                 4 IRREGULARLY FLOODED


                                                 <: IRREGULARLY EXPOSED

                                                 J SUBTIDAL
            Figure 3.4a: Cowardin hierarchy of habitat types for estuarine systems; from Cowardin et al. 1979.
             Nutrient Criteria-Wetlands

              SEEPAGE ZONE
                                            a TEMPORARILY FLOODED
                                            b SEASONALLY FLOODED
                                            c SEMIPERMANENTLY FLOODED
                                            d INTERMrTTTENTLY FLOODED
                                            c PERMANENTLY FLOODED
                                            f SATURATED
                                                                                      HIGH WATER

                                                                                   AVERAGE WATER
                                                                                   LOW WATER
        Figure 3.4b: Cowardin hierarchy of habitat types for Palustrine systems; from Cowardin et al. 1979.

Brinson (1993) has defined a hydrogeomorphic classification system for wetlands based on geomorphic setting,
dominant water source (Figure 3.5), and dominant hydrodynamics (Figure 3.6; http://www.wes.army.mil/el/
wetlands). Seven classes have been described: depressional, lacustrine fringe, tidal fringe, slope, riverine,
mineral soil flats, and organic soil flats (Smith et al. 1995). Also see Hydrogeomorphic Classification in http://

Depressional systems, as the name implies, are located in topographic depressions where surface water can
accumulate. Depression wetlands can be further classified based on presence of inlets or outlets and primary
water source as closed, open/groundwater, or open/surface water subclasses.

Lacustrine fringe wetlands are located along lake shores where the water elevation of the lake determines the
water table of the adjacent wetland. Great Lakes coastal wetlands represent one important region of lacustrine
fringe wetlands. These coastal systems are strongly influenced by coastal forming processes, and, as such,
have been further classified by geomorphic type through various schemes (Jaworski and Raphael 1979, and
others summarized in Michigan Natural Features Inventory 1997). These geomorphic coastal positions
will further influence the predominant source of water and the degree and type of energy regime (riverine
vs. seiche and wave activity). Tidal fringe wetlands occupy a similar position relative to marine coasts and
estuaries, where water level is influenced by sea level. Tidal fringe wetlands can be broken down further
based on salinity  into euhaline vs. mixohaline subclasses. Slope wetlands occur on slopes where groundwater
discharges to the land surface, but typically do not have the capacity for surface water storage (Figure 3.7).
Riverine wetlands are found in floodplains and riparian zones associated with stream channels. Riverine
systems can be broken down based on watershed position (and, thus, hydrologic regime) into tidal, lower
perennial, upper perennial, and nonperennial subclasses. Mineral soil flats are in areas of low topographic
relief (e.g., interfluves, relic lake bottoms, and large floodplain terraces) with precipitation as the main source of
water. The topography of organic soil flats (e.g., peatlands),  in contrast, is controlled by the vertical accretion of
organic matter.
                                         Nutrient Criteria-Wetlands

            Figure 3.5: Dominant water sources to
            wetlands, fromBrinson 1993. Cowardin
            hierarchy of habitat types for estuarine
                        Figure 3.6: Dominant hydrodynamic
                        regimes for wetlands based on flow
                        pattern (Brinson 1993).
                                        «(N» »*»• » «»
                             Figure 3.7: Interaction with break in slope with
                             groundwater inputs to slope wetlands (Brinson 1993).

    The HGM classification system is being further refined to the subclass level for different regions or States
    and classes (Cole et al. 1997, http://www.wes.army.mil/el/wetlands). In addition to the classification factors
    described above, Clairain (2002) suggests using parameters such as the degree of connection between the
    wetland and other surface waters (depressional wetlands), salinity gradients (tidal), degree of slope or channel
    gradient (slope and riverine wetlands), position in the landscape (riverine, slope), and a scaling factor (stream
Nutrient Criteria-Wetlands

order, watershed size or floodplain width for riverine subclasses). In some cases, existing regional schemes
have been used as the basis for subclass definition (e.g., Stewart and Kantrud 1971; Golet and Larson 1974;
Wharton et al. 1982; Weakley and Schafale 1991; Keough et al. 1999).

The HGM classification system has been applied primarily to assess wetland functions related to hydrology,
biological productivity, biogeochemical cycling, and habitat (Smith et al. 1995, http://www.wes.army.mil/
el/wetlands/pdfs/wrpde9.pdf). The same environmental parameters that influence wetland functions also
determine hydrologic characteristics and background water quality, which in turn drive wetland habitat
structure and community composition and the timing of biotic events. Thus, the HGM classification system
can serve as a basis for partitioning variability in reference trophic status and biological condition, as well as
defining temporal strategies for sampling.

If an integrated assessment of aquatic resources within a watershed or region is desired, it may be useful to
consider intercomparability of classification schemes for wetlands, lakes, and riverine systems to promote
cost-effective sampling and ease of interpretation. The HGM approach could integrate readily with a finer
level of classification for lake type because lentic systems are separated out as lacustrine fringe or depressional
wetlands based on lake or pond size and influence of water level on the adjacent wetland. Lacustrine
classification systems for water quality have included geography (climate + bedrock characteristics, Gorham
et al. 1983) or hydrologic setting (Winter 1977; Eilers et al. 1983) as factors for categorization. McKee et al.,
(1992) suggest a modification of Cowardin's system for Great Lakes coastal wetlands incorporating landscape
position (system), depth zone (littoral vs. limnetic subsystems), vegetative or substrate cover (class and
subclass), and modifiers of ecoregions, water level regimes, fish community structure, geomorphic structure,
and human modification. In contrast, the Michigan Natural Features Inventory (1997) categorizes Great Lakes
coastal wetlands by Great Lake, then nine unique geomorphic types within lakes, then vegetative association.

For lotic systems, Brinson et al., (1995) describes an approach to further classify riverine classes into
subclasses based on watershed position and stream size/permanence. This strategy is consistent with current
monitoring efforts to develop stream IBIs (Indices of Biotic Integrity), which typically use stream order as
a surrogate for watershed size in explaining additional background variation in IBI scores (USEPA 1996).
A more detailed classification of stream reach types, based on hydrogeomorphic character, is described by
Rosgen (1996). This classification scheme has been predominantly applied to assessments of channel stability
and restoration options, and not to development of criteria. Gephardt et al., (1990) described a cross-walk
between riparian and wetland classification and description procedures.

It is possible to combine geographically-based classification with hydrogeomorphic and/or habitat-based
approaches.  For example, a scheme could be defined that nests Cowardin (Cowardin et al. 1979) vegetative
cover class within HGM class within ecoregion. Maxwell et al., (1995) have defined a scheme for linking
geographically-based units based on geoclimatic setting (domains => divisions => provinces => sections =>
subsections) to watersheds and subwatersheds, and thus to riverine systems composed of valley segments,
stream reaches, and channel units, or to lacustrine systems composed of lakes, lake depth zones, and lake sites/
habitat types.

Maxwell et al., (1995) also define a series of fundamental hydrogeomorphic criteria for classifying wetlands
based on Brinson (1993) and Winter (1992), including physiography (landscape position), water source,
hydrodynamics, and climate. The first three of these are similar to the HGM classification system (see
summary tables in Keys et al.  1995). Finer scale variation in landforms is also discussed and may be of use in
determining the dominance of different hydrogeomorphic classes of wetlands and associated surface waters
(lakes and rivers). Characteristics and relative advantages and disadvantages of different classification systems
are summarized in Table 2.
                                        Nutrient Criteria-Wetlands                                      3-9


    In order to select wetlands for sampling in a random- or random-stratified design (described in Chapter 4), it
    is important to have a record of wetland locations to choose from, preferably categorized by the classification
    system of interest. For some, but not all portions of the country, wetlands have been mapped from aerial
    photography through the National Wetlands Inventory (NWI) maintained by the U.S. Fish and Wildlife
    Service (http://www.fws.gov/nwi/; Dahl 2005). In other cases, individual States have developed inventories, or
    researchers have developed lists for specific types of wetlands within a given region, e.g., Great Lakes coastal
    wetlands (Herdendorf et al. 1981). In order to sample these mapped wetland areas in a random fashion, it is
    important to have a list of each wetland that occurs within each class and its associated area. A geographic
    information system (GIS) allows one to automatically produce a list of all wetland polygons by type within a
    specified geographic region. Sources of digital information for mapping and/or classifying wetlands in a GIS
    are presented in the Land-Use Characterization for Nutrient and Sediment Risk Assessment Module (http://

    In areas for which digital NWI maps do not yet exist, potential wetland areas can be mapped using GIS tools
    to predict relative wetness (e.g., Phillips 1990) or soil survey maps with hydric soil series can be used. It
    should be noted that in areas in which hydrology has been significantly altered (e.g., through ditching, tiling,
    or construction of urban stormwater systems), areas of potential wetlands could have been removed already.
    Similarly, although there are no current maps of wetlands by hydrogeomorphic class, these could be derived
    through GIS techniques using a combination of wetland coverages, hydrography (adjacency to large lakes
    and rivers), and digital elevation models to derive landforms (mineral and organic soil flats) and/or landscape
    position (slope and depressional wetlands).


    Very few studies to verify classification systems for wetland nutrient monitoring have been completed,
    although a number of monitoring strategies have been implemented based on pre-selected strata. Monitoring
    efforts to develop or assess biological criteria generally have used a combination of geographic region and
    hydrogeomorphic class or subclass (e.g., Cole et al. 1997; Bennett 1999; Apfelbeck 1999; Michigan Natural
    Features Inventory 1997). Analysis of plant associations has been used to derive empirical classifications based
    on factors such as landscape position, water source, climate, bedrock, and sediment hydraulic conductivity
    (Weakley and Schafale 1991; Nicholson 1995; Halsey et al. 1997; Michigan Natural Features Inventory 1997).
    Only one case of classification based on wetland macroinvertebrate composition was found. For Australian
    wetlands, wetland classes grouped by macroinvertebrate communities were distinguished by water chemistry
    extremes (low pH, high salinity), degree of nutrient enrichment, and water color (Growns et al. 1992).

    In some cases (e.g., northern peatlands) classification criteria derived on the basis of plant associations are
    less powerful in discriminating among nutrient regimes (e.g., Nicholson 1995); this may be particularly
    true where variation in vegetation type is related to differences in major ion chemistry and pH, rather than
    nutrients.  The same is true in southern pocosins, where short and tall pocosins differ in seasonal hydrology
    but not soil chemistry. However, when contrasting pocosins and swamp forests, soil nutrients differed strongly
    (Bridgham and Richardson 1993). For some potential indicators of nutrient status such as vegetation nitrogen
    to phosphorus ratios, indicator thresholds will be consistent across species (Koerselman and Meuleman 1996),
    while response thresholds for other indicators of plant nutrient status vary across functional plant groupings
    with different life history strategies. These differences may indicate potential differences in sensitivity to
    excess nutrient loading (McJannet et al. 1995). Thus, vegetation community types are not always a good
    predictor of background nutrient concentrations (reference condition) or sensitivity to nutrient loading.

    Sensitivity to nutrient loading (as evidenced by differences in nutrient cycling and availability) may also be
3-10                                     Nutrient Criteria-Wetlands

related to differences in hydroperiod among wetlands. Wetland mesocosms exposed to pulse discharges had
higher nutrient loss from the water column than those exposed to continuous flow regimes (Busnardo et al.
1992). Depending on the predominant mechanism for nutrient loss (e.g., plant uptake versus denitrification),
nutrient-controlled primary production could be either stimulated or reduced. Mineralization rates of carbon,
nitrogen, and phosphorus differ significantly among soils from northern Minnesota wetlands, related to an
ombrotrophic to minerotrophic gradient (i.e., degree of groundwater influence), and aeration status (Bridgham
et al. 1998).

In general, very few definitive tests of alternative classification schemes for wetlands are available with
respect to describing reference condition for either nutrient criteria or biocriteria. However, evidence from the
literature suggests that in many cases both geographic factors (e.g., climate, geologic setting) and landscape
setting (hydrogeomorphic type) are expected to affect water quality and biotic communities.


Classification strategies for nutrient criteria development should incorporate  factors affecting background
nutrient levels and wetland sensitivity to nutrient loading at several spatial scales.

        • Classification of physiographic regions eliminates background variation in lithology and soil
          texture (affecting background nutrient levels and sorption capacity), in climate (affecting
          seasonality, productivity,  decomposition, and peat formation), and in landforms, which
          determines the predominance of different hydrogeomorphic classes.

        • Classification by hydrogeomorphic class reduces background variation in predominant water
          and nutrient sources, water depth and dynamics, hydraulic retention time, assimilative
          capacity, and interactions with other surface water types (Table 3).

        • Classification by water depth and duration (which may or may not be incorporated
          into hydrogeomorphic classes) helps to explain variation in internal nutrient cycling,
          dissolved oxygen level and variation, and the ability of wetlands to support some higher
          trophic levels such as fish and amphibians.

        • Classification by vegetation type or zone, whether to inform site selection or to etermine
          sampling strata within a site, helps to explain background variation in predominant primary
          producer form (which will affect endpoint selection), as well as turnover and growth rates
          (which will affect rapidity of response to nutrient loadings).

In general, the choice of specific alternatives among the classification schemes listed above depends on
their intrinsic value as well as practical considerations, e.g., whether a classification scheme is available in
mapped digital form or can be readily derived from existing map layers, whether a hydrogeomorphic or other
classification scheme has been refined for a particular region and wetland type, and whether classification
schemes are already in use for monitoring and assessment of other waterbody types in a state or region.
Revisiting classification decisions once data from a sufficient number of sites have been sampled may be useful
to ensure the original classification was correct.
                                         Nutrient Criteria-Wetlands                                     3-11

Bailey's ecoregions

Omernik ecoregions

Ecological units
(Maxwell et al. 1995)


Rosgen channel types

Anderson land-cover

Circular 39 classes

National Wetland





at class level;
at subclass








Yes - limited










Level I
Level II

Level I
Level II
Level III

Other modifiers
Formation class
Only natural
attributes included

Digital maps

Digital maps

Digital maps

Specific for

differences in
hydrologic regime
for riverine

Common basis for
Popular recognition

Digital maps
available for much
of nation (but
smallest wetlands

Consistency across
terrestrial and
aquatic systems

Terrestrial basis

Untested for wetlands

No hydrology
Combines land-use
with natural attributes

Untested for most
No hydrology
Greater number of
strata and units than
for ecoregions

Untested for wetlands

Subclasses not
comparable across
different regions

More focused on
instream channel
form than riparian

Riverine only
Not mapped
Not functionally based

Mixture of criteria
used to distinguish

Not mapped
Inconsistencies in
mapping water quality

Limited consideration
of hydrogeomorphic
Not functionally based

No digital maps

Taxa specific

Could form first strata for any
of the schemes below ecological

Could form first strata for any
of the schemes below ecological

Could form first strata for any
of the schemes below ecological

Ties to classification schemes
already defined within
hydrogeomorphic types
Intermediate strata between
geographic and habitat-scale

Intermediate strata between
hydro-geomorphic type and

Cross-walk w NWI system

Strata below geographic
but contains mixture of
hydrogeomorphic type and habitat

Strata below geographic
Hydrogeomorphic class could be
improved by link w HGM system

Could be used as lowest level
within other schemes

             Table 2: Comparison of landscape and wetland classification schemes.
Nutrient Criteria-Wetlands

Predominant Nutrient

Landscape Position
Hydrologic Regime

Hydraulic Retention

Nutrient Assimilation

Vegetation Growth

Top Trophic Level

Important Fish/
Recreational Use
Drinking Water Source







Saturated, Little
Standing Water


High Sorption



(P articulate
and Dissolved),
Surface and

Depth and
Duration Vary
from Saturated
to Temporary
to Seasonal to
to Permanent
Varies With
High Sorption,
Plant Uptake,
Sediment Storage
Varies With Zone
And Duration of



(P articulate),
Overbank Flooding
(P articulate,
Adjacent to Rivers
Depth, Duration
Vary With River
Flooding Regime




This chapter provides technical guidance on designing effective sampling programs for State wetland water
quality monitoring programs. EPA recommends that States begin wetland monitoring programs to collect
water quality and biological data in order to characterize the condition of existing wetlands as they develop
nutrient criteria that will protect their wetlands. The best monitoring programs are designed to assess wetland
conditions with statistical rigor while maximizing available resources.

At the broadest level, monitoring data should:

        1.  Detect and characterize the condition of existing wetlands.
        2.  Describe whether wetland conditions are improving, degrading, or staying the same.
        3.  Define seasonal patterns, impairments, and deviations in status of wetland conditions.

Water quality monitoring programs should collect a sufficient number of samples over time and space to
identify changes in system condition or estimate average conditions with statistical rigor. Three approaches to
study design for assessing water quality and biological and ecological condition, and identifying degradation
in wetlands are described in this chapter.  Specific issues to consider in designing monitoring programs for
wetland systems are also discussed in this chapter. The study designs presented here can be tailored to fit the
goals of specific monitoring programs.

The three approaches described below (Section 4.3) (probabilistic sampling, targeted/tiered, and Before/After-
Control/Impact [BACI]), present study designs that allow one to obtain a significant amount of information
with relatively minimal effort. Probabilistic sampling begins with a large-scale, random monitoring design
that is reduced as the wetland system conditions are characterized.  This approach is used to find the average
condition of each wetland class in a specific region. Probabilistic sampling design is frequently used for
new large-scale monitoring programs at the State and Federal level (e.g., Environmental Monitoring and
Assessment Program (EMAP), Regional Environmental Monitoring and Assessment Program (REMAP),
State programs [e.g., Maine, Montana, Wisconsin]). The tiered or targeted approach to monitoring begins
with coarse screening and proceeds to more detailed monitoring protocols as impaired and high-risk systems
are identified and targeted for further investigation. Targeted sampling design provides a triage approach to
more thoroughly assess condition and diagnose stressors in wetland systems in need of restoration, protection,
and intensive management. Several State  pilot projects use this method or a modification of this method for
wetland assessment (e.g., Florida, Ohio, Oregon, and Minnesota). The synoptic approach described in Kentula
et al., (1993) uses a modified targeted sampling design. The BACI design and its modifications are frequently
used to assess the success of restoration efforts or other management experiments. BACI design allows for
comparisons in similar systems over  time to determine the rate of change in relation to the management
activity, e.g., to assess the success of a wetland hydrologic restoration. The BACI design, in particular, is
included to assist States in evaluating ongoing management actions, and may provide less statistical rigor if
adopted as a general monitoring program design. This design, however, is of considerable value in assessing
restoration success and has been included at the request of States with ongoing wetland restoration. Detenbeck
et al., (1996) used BACI design for monitoring water quality of wetlands in the Minneapolis/St. Paul,
Minnesota metro area.

Monitoring programs should be designed to describe what the current conditions are and to answer under what
conditions impairment may occur. A well-designed monitoring program can contribute to determining those

Sampling design is dependent on the management question being asked. Sampling efforts should be designed
                                        Nutrient Criteria-Wetlands                                     4-1

    to collect information that will answer the management question. For example, probabilistic sampling might
    be good for ambient (synoptic) monitoring programs, BACI for evaluating management actions such as
    restoration, and targeted sampling/stratified and random sampling for developing Index of Biotic Integrity
    (IBIs) or nutrient criteria thresholds. In practice, some State programs likely will need to use a combination of


    Clearly defining the question being asked (identifying the hypothesis) encourages the use of appropriate
    statistical analyses, reduces the occurrence of Type I (false positive) errors, and increases the efficient
    use of management resources (Suter 1993; Leibowitz et al. 1992; Kentula et al. 1993). Beginning a study
    or monitoring program with carefully defined questions and objectives helps to identify the statistical
    analyses most appropriate for the study and reduces the chance that statistical assumptions will be violated.
    Management resources are optimized because resources are directed at monitoring that which is most likely
    to answer management questions. In addition, defining the specific hypotheses to be tested, carefully selecting
    reference sites,  and identifying the most useful sampling interval can help reduce the uncertainty associated
    with the results of any sampling design and further conserve management resources (Kentula et al. 1993).
    Protecting  or improving the quality of a wetland system often depends on the  ability of the monitoring
    program to identify cause-response relationships, for example, the relationship of nutrient concentration
    (causal variable) to nutrient content of vegetation or vegetation biomass (response variable). Cause-response
    relationships can be identified using large sample sizes and systems that span the gradient (low to high) of
    wetland quality. All ranges of response should be observed along the causal gradient from minimally disturbed
    to high levels of human disturbance.

    Monitoring efforts often are prioritized to best utilize limited resources. For example, the Oregon case study
    chose not to monitor depressional wetlands due to funding constraints. They further tested the degree of
    independence of selected sites (and thus the need to monitor all of those sites)  using cluster analysis and other
    statistical tests (http://www.epa.gov/owow/wetlands/bawwg/case/or.html). Frequency of monitoring should
    be determined by the management question being asked and the intensity of monitoring necessary to collect
    enough information to answer the question. In addition, monitoring should identify the watershed level
    activities that are likely to result in ecological degradation of wetland systems (Suter et al. 1993).

    Site selection is one of many important tasks in developing a monitoring program (Kentula et al. 1993). Site
    selection for a monitoring program is based on the need to sample a sufficiently large number of wetlands to
    establish the range of wetland quality in a specific regional setting. Wetland monitoring frequently includes an
    analysis of both watershed/landscape characteristics and wetland specific characteristics (Kentula et al. 1993;
    Leibowitz  et al. 1992). Therefore, wetland sampling sites should be selected based on land use in the region
    so that watersheds range from minimally impaired with few expected stressors to high levels of development
    (e.g., agriculture, forestry, or urban) with multiple expected stressors (see the Land-Use Characterization
    for Nutrient and Sediment Risk Assessment). There is often a lag in time between the causal stress and the
    response in the wetland system. This time lag between stress and response and the duration of this lag depends
    on many factors, including the type of stressor, climate, and system hydrology; these factors should be
    considered when selecting sites to establish the range of wetland quality within a region.

    The synoptic approach described in Liebowitz et al., (1992) provides a method of rapid assessment of
    wetlands at the regional and watershed levels that can help identify the range of wetland quality within  a
    region. Liebowitz et al., (1992) recommend an initial assessment for site selection based on current knowledge
    of watershed and landscape level features; modification of such an assessment can be made as more data
4-2                                      Nutrient Criteria-Wetlands

are collected. Assessing watershed characteristics through aerial photography and the use of geographical
information systems (GIS) linked to natural resource and land-use databases can aid in identifying reference
and degraded systems (see the Land-Use Characterization for Nutrient and Sediment Risk Assessment);
Johnston et al. 1988; 1990; Gwin et al. 1999; Palik et al. 2000; Brown and Vivas 2004).  Some examples of
watershed characteristics which can be evaluated using GIS and aerial photography include land use, land
cover (including riparian vegetation),  soils, bedrock, hydrography, and infrastructure (e.g., roads or railroads).
Changes in point sources can be monitored through the NPDES permit program (USEPA 2000). Changes in
nonpoint sources can be evaluated through the identification and tracking of wetland loss and/or degradation,
increased residential development, urbanization, increased tree harvesting, shifts to more intensive
agriculture with greater fertilizer use  or increases in livestock numbers, and other land  use changes.  Local
planning agencies should be informed of the risk of increased anthropogenic stress and encouraged to guide
development accordingly.

The term "reference" in this document refers to those systems that are least impaired by anthropogenic
effects. The use of the term reference  is confusing because of the different meanings that are currently in use
in different classification methods, particularly its use in hydrogeomorphic (HGM) wetland classification. A
discussion of the term reference and its multiple meanings is provided in Chapter 3.

Watersheds with little or no development that receive minimal anthropogenic inputs could potentially
contain wetlands that may serve as minimally impaired reference sites. Watersheds with a high percentage
of the drainage basin occupied by urban areas, agricultural land, and altered hydrology are likely to contain
wetlands that are impaired or could potentially be considered "at risk" for developing problems. Wetland
loss in the landscape also should be considered when assessing watershed characteristics for reference
wetland identification. Biodiversity can become impoverished due to wetland fragmentation or decreases
in regional wetland density even in the absence of site-specific land-use activities. Reference wetlands may
be more difficult to locate if fragmentation of wetland habitats is significant and may no longer represent
the biodiversity of minimally disturbed wetlands in the region. The continued high rate of wetland loss in
most States dictates that multiple reference sites be selected to ensure some consistency in reference  sites for
multiple year sampling programs (Liebowitz et al. 1992; Kentula et al. 1993). Once the watershed level has
been considered, a more site-specific investigation can be initiated to better assess wetland condition.

The ideal reference site will have similar soils, vegetation, hydrologic regime, and landscape setting to other
wetlands in the region (Adamus 1992; Liebowitz et al. 1992; Kentula et al. 1993; Detenbeck et al. 1996).
Classification of wetlands, as discussed in Chapter 3, may aid in identifying appropriate reference wetlands for
specific regions and wetland types. Wetland classification should be supplemented with information on wetland
hydroperiod to assure that the selected reference wetlands are truly representative of wetlands in the  region,
class, or subclass of interest. Reference wetlands may not be available for all wetland classes. In that case,
data from systems that are as close as possible to the assumed unimpaired state of wetlands in the wetland
class of interest should be sought from States within the same geologic province. Development of a conceptual
reference may be important if appropriate reference sites cannot be found in the local region or geologic
province. Techniques for defining a conceptual reference are discussed at some length in Harris et al., (1995),
Trexler (1995), and Toth et al., (1995).

Reference wetlands  should be selected based on low levels of human alteration in their watersheds (Liebowitz
et al. 1992; Kentula  et al. 1993; USEPA 2000). Selecting reference wetlands usually involves assessment of
land-use  within watersheds and visits to individual wetland systems to ground-truth expected land-use and
check for unsuspected impacts. Ground-truthing visits to reference wetlands are crucial for identification of
ecological impairment that may not be apparent from land-use and local habitat conditions.  Again, sufficient
sample size is important to characterize the range of conditions that can be expected in the least impacted
systems of the region (Detenbeck et al. 1996). Reference wetlands should be identified for each ecoregion or
                                        Nutrient Criteria-Wetlands                                      4-3

    geological province in the State lands and then characterized with respect to ecological integrity. A minimum
    of three low impact reference systems is recommended for each wetland class for statistical analyses. However,
    power analysis can be performed to determine the degree of replication necessary to detect an impact to the
    systems being investigated (Detenbeck et al. 1996; Urquhart et al. 1998). Highest priority should be given to
    identifying reference systems for those wetland types considered to be at the greatest risk from anthropogenic

    Sampling may be targeted to the periods when effects are most likely to be detected - the index period. The
    appropriate index period should be defined by what the investigator is trying to investigate and what taxonomic
    assemblage or parameters are being used for that investigation (Barbour et al.  1999). For example, increased
    nutrient concentrations and sedimentation from non-point sources may occur following periods of high runoff
    during spring and fall, while point sources of nutrient pollutants may cause plankton blooms and/or increased
    water and soil nutrient concentrations in wetland pools during times of low rainfall. Hence, different index
    periods may be needed to detect effects from point source and nonpoint source nutrients, respectively. Each
    taxonomic assemblage studied also should have an appropriate index period—usually in the growing season
    (see assemblage methods in the Maine case study: http://www.epa.gov/waterscience/criteria/wetlands/).

    The index period window may be early in the growing season for amphibians and algae. Other assemblages,
    such as vegetation and birds, may benefit from a different sampling window for the index period; see the
    assemblage specific modules for recommendations. Once wetland condition has been characterized, one-
    time annual sampling during the appropriate index period may be adequate for multiple year monitoring of
    indicators of nutrient status, designated use, and biotic integrity. However, criteria and ecological indicator
    development may benefit from more frequent sampling to define conditions that relate to the stressor or
    perturbor of interest (Karr and Chu 1999; Stevenson 1996; Stevenson 1997). Regardless of the frequency of
    sampling, selection of index periods and critical review of the data gathered and analyzed should be done  to
    scientifically validate the site characterization and index periods for data collection.

    Ideally, water quality monitoring programs produce long-term data sets compiled over multiple years
    to capture the natural, seasonal, and year-to-year variations in biological communities and constituent
    concentrations (e.g., Tate 1990; Dodds et al. 1997; McCormick et al. 1999; Craft 2001; Craft et al. 2003; Zheng
    et al. 2004). Multiple-year data sets can be analyzed with statistical rigor to identify the effects of seasonality
    and variable hydrology. Once the pattern of natural variation has been described, the data can be analyzed to
    determine the ecological state of the wetland. Long-term data sets have also been important in influencing
    management decisions about wetlands, most notably in the Everglades, where long-term data sets have induced
    Federal, State, and Authorized Tribal actions for conservation and restoration of the largest wetland system in
    the U.S. (see Davis and Ogden 1994; Everglades Interim Report, South Florida Water Management District
    [SFWMD 1999]; Everglades Consolidated Report [SFWMD 2000, 2001]; 1994 Everglades Forever Act,
    Florida Statute § 373.4592).

    In spite of the documented value of long-term data sets, there is a tendency to intensively study a wetland
    for one year before and one year after treatment. A more cost-effective approach may be to measure only the
    indices most directly related to the stressor of interest (i.e.,  those parameters or indicators that provide the
    best information to answer the specific management question), but to double or triple the monitoring period.
    Multiple years (two or more) of data are often needed to identify the effects of years with extreme climatic
    or hydrologic conditions. Comparisons over time between reference and at risk or degraded systems can help
    describe biological response and annual patterns in the presence of changing climatic conditions. Multi-year
    data sets also can help describe regional trends. Flooding or drought may significantly affect wetland biological
    communities and the concentrations of water column and soil constituents. Effects of uncommon climatic
    events can be characterized to discern the overall effect of management actions (e.g., nutrient reduction, water
    diversion) if several years  of data are available to identify the long-term trends.
4-4                                      Nutrient Criteria-Wetlands

At the very minimum, two years of data before and after specific management actions, but preferably three
or more each, are recommended to evaluate the cost-effectiveness of management actions with some degree
of certainty (USEPA 2000). If funds are limited, restricting sampling frequency and/or numbers of indices
analyzed should be considered to preserve a longer-term data set. Reducing sampling frequency or numbers of
parameters measured will allow for effectiveness of management approaches to be assessed against the high
annual variability that is common in most wetland systems. Wetlands with high hydrological variation from
year to year may benefit from more years of sampling both before and after specific management activities to
identify the effects of the natural hydrologic variability (Kadlec and Knight 1996).

Estimates of cause-response relationships, nutrient and biological conditions in reference systems, and wetland
conditions in a region are based on sampling; hence, precision should be assessed. Precision is defined as
the "measure of the degree of agreement among the replicate analyses of a sample, usually expressed as the
standard deviation" (APHA 1999). Determining precision of measurements for one-time assessments from
single samples in a wetland is often important. The variation associated with one-time assessments from single
samples can be determined by re-sampling a specific number of wetlands during the survey. Measurement
variation among replicate samples then can be used to establish the expected variation for one-time assessment
of single samples. Re-sampling does not establish the precision of the assessment process, but rather identifies
the precision of an individual measurement  (Kentula et al. 1993).

Re-sampling frequency is often conducted for one wetland site in every block of 10 sites. However,
investigators should adhere to the objectives of re-sampling (often considered an essential element of Quality
Assurance/Quality Control (QA/QC)) to establish an assessment of the variation in a one-time/sample
assessment. Often,  more than one in 10 samples should be replicated in monitoring programs to  provide a
reliable estimate of measurement precision (Barbour et al. 1999). The reader should understand that this is a
very brief description of the concerns about precision, and that any monitoring program or study involving
monitoring should include  consultation with a professional statistician before the program begins and regularly
during the course of the monitoring program to assure statistical rigor.


The following sections discuss three different approaches to sampling design, probabilistic, targeted, and
BACI.  These approaches have advantages and disadvantages that under different circumstances warrant the
choice  of one approach over the other (Table 4). The decision as to the best approach for sample design in a
new monitoring program should be made by the water quality resource manager or management team after
careful consideration of the different approaches. For example, justification of a dose-response relationship is
confounded by lack of randomization and replication and should be considered in choosing a sampling design
for a monitoring program.

Probabilistic sampling - a sampling process wherein randomness is requisite (Hayek 1994) - can be used to
characterize the status of water quality conditions and biotic integrity in a region's wetland system. This type
of sampling design is used  to describe the average conditions of a wetland population, identify the variability
among sampled wetlands, and help determine the range of wetland system conditions in a region. Data
collected from a probabilistic random sample design generally will be characteristic of the dominant class or
type of wetland in the region, but rare wetlands may be under-represented or absent from the probabilistically
sampled wetlands. Additional sampling sites may need to be added to precisely characterize the  complete
range of wetland conditions and types in the region.
                                        Nutrient Criteria-Wetlands                                      4-5

    Probabilistic designs are often modified by stratification (such as classification). Stratified random sampling
    is a type of probabilistic sampling where a target population is divided into relatively homogenous groups or
    classes (strata) prior to sampling based on factors that influence variability in that population (Hayek 1994).
    Stratification by wetland size and class or types ensures more complete information about different types of
    wetlands within a region. Sample statistics from random selection alone would be most characteristic of the
    dominant wetland type in a region if the population of wetlands is not stratified.

    Many State 305(b) and watershed monitoring programs utilize stratified random sampling designs, and we
    will further discuss this type of probabilistic sampling. Pilot projects in Maine, Montana, and Wisconsin all
    use stratified random sampling design. Details of these monitoring designs can be found in the Case Studies
    Module #14 and on the Web at http://www.epa.gov/waterscience/criteria/wetlands/index.html.

    Stratification is based on identifying wetland systems in a region (or watershed) and then selecting an
    appropriate sample of systems from the defined population. The determination of an appropriate sample
    population usually is dependent on the management questions being asked. A sample population of isolated
    depressional wetlands could be identified as a single stratum, but investigations of these wetlands would not
    provide any information on riparian wetlands in the same region. If the goal of the monitoring program is to
    identify wetland condition for all wetland classes within a region, then a sample population of wetlands should
    be randomly selected from all wetlands within each class. In practice, most State programs stratify random
    populations by size, wetland class  (see Chapter 3), and landscape characteristics or location (see http://www.
    epa.gov/waterscience/criteria/wetlands/, http://www.epa.gov/waterscience/criteria/wetlands/17LandUse.pdf).

    Once the wetlands for each stratum have been identified, the list of wetlands can be used to select a spatially-
    balanced stratified random sample. Spatial-balance will ensure spatial coverage over the assessment region,
    usually increase the types of wetlands sampled (assuming classes of wetlands vary spatially), and reduce
    spatial autocorrelation among the sampled wetlands. For example, EMAP implements spatially-balanced
    samples using Generalized Random Tessellation Stratified (GRTS) designs applied to GIS coverages of
    wetlands within the assessment region. GRTS using a hierarchical grid randomization process to ensure the
    sites are spatially distributed (Paulsen et al.  1991; Stevens and Olsen 2004). Estimates of ecological conditions
    from these kinds of modified probabilistic sampling designs can be used to characterize the water quality
    conditions  and biological integrity of wetland systems in a region, and over time, to  distinguish trends  in
    ecological condition within a region. (See http://www.epa.gov/owow/wetlands/bawwg/case/mtdev.html and

    A targeted approach to sampling design may be more appropriate when resources are limited (Stern 2004). The
    example of targeted sampling described here involves defining a gradient of impairment. Once the gradient has
    been defined and systems have been placed in categories of impairment, investigators focus the greatest efforts
    on identifying and characterizing wetland systems or sites likely to be impacted by anthropogenic stressors,
    and on relatively undisturbed wetland systems or sites (see Identifying and Characterizing Reference Systems,
    Chapter 3), that can serve as regional, sub-regional,  or watershed examples of natural biological integrity.
    Florida Department of Environmental Protection (FDEP) uses a targeted sampling design for developing
    thresholds of impairment with macroinvertebrates (http://www.epa.gov/owow/wetlands/bawwg/case/fl2.html).
    Choosing sampling stations that best allow comparison of ecological integrity at reference wetland sites of
    known condition can conserve financial resources. A sampling design that tests specific hypotheses (e.g., the
    FDEP study tested the effect of elevated water column phosphorus on macroinvertebrate species richness)
    generally can be analyzed with statistical rigor and can conserve resources by answering specific questions.
    Furthermore, identification of systems with problems and reference conditions eliminates the need for
    selecting a random sample of the population for monitoring.
4-6                                      Nutrient Criteria-Wetlands

Targeted sampling assumes some knowledge of the systems sampled (Stern 2004; Kentula et al. 1993). Systems
based on independent variables with evidence of degradation are compared to reference systems that are
similar in their physical structure (i.e., in the same class of wetlands). Wetland systems should be viewed along
a continuum from reference to degraded. An impaired or degraded wetland is a system in which anthropogenic
impacts exceed acceptable levels or interfere with beneficial uses. Comparison of the monitoring data to that
collected from reference wetlands will allow characterization of the sampled systems. Wetlands identified as
"at risk" should be evaluated through a sampling program to characterize the degree of degradation. Once
characterized, the wetlands should be placed in one of the following categories:

        1.  Degraded wetlands—wetlands in which the level of anthropogenic perturbance interferes
           with designated uses.

        2.  High-risk wetlands—wetlands where anthropogenic stress is high but does not significantly
           impair designated uses. In high risk systems, impairment is prevented by one or a few
           factors that could be changed by human actions, though characteristics of ecological integrity are
           already marginal.

        3.  Low-risk wetlands—wetlands where many factors prevent impairment, stressors are maintained
           below problem levels, and/or no development is contemplated that would change these conditions.

        4.  Reference wetlands—wetlands where the ecological characteristics most closely represent the
           pristine or minimally impaired  condition.

Once wetland systems have been classified based on their physical structure (see Chapter 3) and placed into the
above categories, specific wetlands need to be selected for monitoring. At this point, randomness is introduced;
wetlands should be randomly selected within each class and risk category for monitoring. An excellent
example of categorizing wetlands in this manner is given in the Ohio Environmental Protection Agency's (OH
EPA) case study, available at: http://www.epa.gov/owow/wetlands/bawwg/case/ohl.html. They used the Ohio
Rapid Assessment Method to categorize wetlands by degree of impairment. The Minnesota Pollution Control
Agency (MPCA) also used a targeted design for monitoring wetlands (http://www.epa.gov/owow/wetlands/
bawwg/case/mnl.html). They used the best professional judgment of local resource managers to identify
reference sites and those with known impairment from identified stressors (agriculture and stormwater runoff).

Targeted sampling design involves monitoring identified degraded systems and comparable reference systems
most intensively. Low risk systems are monitored less frequently (after initial identification) unless changes in
the watershed indicate an increased risk of degradation.

Activities surrounding impaired wetland systems may be used to help identify which actions negatively affect
wetlands, and therefore may initiate more intensive monitoring of at-risk wetlands. Monitoring should focus on
factors likely to identify ecological degradation and anthropogenic stress and on any actions that might alter
those factors. State water quality agencies should encourage adoption of local watershed protection plans to
minimize ecological degradation of natural wetland systems.  Development plans in the watershed should be
evaluated to identify potential future stressors. Ecological degradation often gradually increases due to many
growing sources of anthropogenic stress. Hence, frequent monitoring may be warranted for high-risk wetlands
if sufficient resources remain after meeting the needs of degraded wetlands. Whenever development plans
appear likely to alter factors that maintain ecological integrity in a high-risk wetland (e.g., vegetated buffer
zones), monitoring should be initiated at a higher sampling frequency in order to enhance the  understanding of
baseline conditions (USEPA 2000).

An ideal before/after impact survey has  several features: (1) the type of impact, time of impact, and place of
occurrence should be known in advance; (2) the impact should not have occurred yet; and, (3) control areas
                                        Nutrient Criteria-Wetlands                                     4-7

    should be available (Green 1979). The first feature allows the surveys to be efficiently planned to account for
    the probable change in the environment. The second feature allows a baseline study to be established and
    extended as needed. The last feature allows the surveyor to distinguish between temporal effects unrelated
    to the impact and changes related to the impact. In practice however, advance knowledge of specific impacts
    is rare, and the ideal impact survey is rarely conducted. BACI designs modified to monitor impacts during or
    after their occurrence still can provide information, but there is an increase in the uncertainty associated with
    the results and the likelihood of finding a statistically significant change due to the impact is less probable.
    In addition, other aspects of survey design are dependent on the study objectives, e.g., the sampling interval,
    the length of time the survey is conducted (i.e., sampling for acute versus chronic effects), and the statistical
    analyses appropriate  for analyzing the data (Suter 1993).

    The best interval for  sampling is determined by the  objectives of the study (Kentula et al. 1993). If the
    objective is to detect  changes in trends (e.g., regular monitoring for detection of changes in water quality
    or biotic integrity), regularly spaced intervals are preferred because the analysis is easier. On the other
    hand, if the objective is to assess differences before  and after impact, then samples at random time points
    are advantageous.  Random sample intervals reduce  the likelihood that cyclic differences unforeseen by the
    sampler will influence the size of the difference before and after the impact. For example, surveys taken
    every summer for a number of years before and after a clear-cut may show little difference in system quality;
    however, differences may exist that can only be detected in the winter and therefore may go undetected if
    sampling occurs only during summer.

    The simplest impact survey design involves taking a single survey before and after the impact event (Green
    1979). This type of design has the obvious pitfall that there may be no relationship between the observed event
    and the changes in the response variable—the change may be entirely coincidental. This pitfall is addressed
    in BACI design by comparing before and after impact data to data collected from a similar control system
    nearby. Data are collected before and after a potential disturbance in two areas (treatment and a control), with
    measurements on biological and environmental variables in all combinations of time and area (Green 1979).
    We will use a  clear-cut adjacent to a wetland as an example to illustrate the BACI design. The sampling design
    is developed to identify the effects of clear-cutting on adjacent wetland systems. In the simplest BACI design,
    two wetlands would be sampled. One wetland would be adjacent to the clear-cut (the treatment wetland);
    the second wetland would be adjacent to a control site that is not clear-cut. The control site should have
    characteristics (soil, vegetation, structure, functions) similar to the treatment wetland and is exposed to climate
    and weather similar to the first wetland. Both wetlands are sampled at the same time points before the clear-cut
    occurs and at the same time point after the clear-cut takes place. This design is technically known as an area-
    by-time factorial design. Evidence of an impact is found by comparing the control site samples (before and
    after) with the treatment site before and after samples. Area-by-time factorial design allows for both natural
    wetland-to-wetland variation and coincidental time  effects. If there is no effect of the clear-cut, then change in
    system quality between the two time points  should be the same. If there is an effect of the clear-cut, the change
    in system quality between the two time points should be different.

    There are some potential problems with BACI design. First, because the control and impact sites are not
    randomly assigned, observed differences between sites may be related solely to some other factor that differs
    between the two sites. One could argue that it  is unfair to ascribe the effect to the impact (Hurlbert 1984;
    Underwood 1991). However, as pointed out by Stewart-Oaten et al., (1986), the survey is concerned about
    a particular impact in a particular place, not in the average of the  impact when replicated in many different
    locations. Consequently, it may be possible to detect a difference between these two specific sites. Even so, if
    there are no randomized replicate treatments, the results of the study cannot be generalized to similar events
    at different wetlands. In any case, the likelihood that the differences between sites are due to factors other than
    the impact can be reduced by monitoring several control sites (Underwood 1991) because multiple control sites
    provide some information about potential effects of other factors.
4-8                                      Nutrient Criteria-Wetlands

The second and more serious concern with the simple Before-After design with a single sampling point before
and after the impact is that it fails to recognize that there may be natural fluctuations in the characteristic
of interest that are unrelated to any impact (Hurlbert 1984; Stewart-Oaten 1986). Single samples before and
after impact would be sufficient to detect the effects of the impact if there were no natural fluctuations over
time. However, if the population also has natural fluctuations over and above the long-term average, then
it is impossible to distinguish between cases where there is no effect from cases where there is an impact.
Consequently, measured differences in system quality may be artifacts of the sampling dates and natural
fluctuations may obscure differences or lead one to believe differences are present when they are not.

The simple BACI design was extended by Stewart-Oaten et al., (1986) by pairing surveys at several selected
time points before and after the impact to help resolve the issue of psuedoreplication (Hulbert 1984). This
modification of the BACI design is referred to as BACI-PS (Before-After, Control-Impact Paired Series
design). The selected sites are measured at the same time points.  The rationale behind this paired design is
that repeated sampling before the impact gives an indication of the pattern of differences of potential change
between the two sites. BACI-PS study design provides information both on the mean difference in the wetland
system quality before and after impact and on the natural variability of the system quality measurements.
The resource manager has detected an effect if the changes in the mean difference are large relative to natural
variability. Considerations for sampling at either random or regularly spaced intervals also apply here.
Replication of samples should also be included if resources allow in order to improve certainty of analytical

Violation of the BACI assumptions may invalidate conclusions drawn from the data. Enough data should
be collected before the impact to identify the trends in the communities of each sampling site if the BACI
assumptions are to be met. Clearly defining the objectives of the  study and identifying a statistically testable
model of the relationships the  investigator is studying can help resolve these issues (Suter 1993).

The designs described above are suitable for detecting longer-term chronic effects in the mean level of the
variable of interest. However, the impact may have an acute effect (i.e., effects only last for a short while)
or may change the variability in response (e.g., seasonal changes become more pronounced) in some cases.
The sampling schedule can be modified so that it occurs at two temporal scales (enhanced BACI-PS design)
that encompass both acute and chronic effects (Underwood 1991). The modified temporal design introduces
randomization by randomly choosing sampling occasions in two periods (Before and After) in the control or
impacted sites. The two temporal scales (sampling periods vs. sampling occasions) allow the detection of a
change in mean and of a change in variability after impact. For example, groups of surveys could be conducted
every year with five surveys one week apart randomly located within each group. The analysis of such a design
is presented in Underwood (1991). Again, multiple control sites should be used to counter the argument that
detected differences are specific to the sampled site. The September 2000 issue of the Journal of Agricultural,
Biological, and Environmental Statistics discusses many of the advantages and disadvantages of the BACI
design and provides several examples of appropriate statistical analyses for evaluation of BACI studies.
                                        Nutrient Criteria-Wetlands                                     4-9

    4.4  SUMMARY

    State monitoring programs should be designed to assess wetland condition with statistical rigor while
    maximizing available management resources. The three approaches described in this module—probabilistic
    sampling, targeted/tiered approach, and BACI (Before/After, Control/Impact)—present study designs that
    allow one to obtain a significant amount of information for statistical analyses. The sampling design selected
    for a monitoring program should depend on the management question being asked. Sampling efforts should
    be designed to collect information that will answer management questions in a way that will allow robust
    statistical analysis. In addition, site selection, characterization of reference sites or systems, and identification
    of appropriate index periods are all of particular concern when selecting an appropriate  sampling design.
    Careful selection of sampling design will allow the best use of financial resources and will result in the
    collection of high quality data for evaluation of the wetland resources of a State. Examples of different sampling
    designs currently in use for State wetland monitoring are described in the Case Study Module #14 on the Web
    site http://www.epa.gov/waterscience/criteria/wetlands/. Well-designed monitoring programs tend to produce
    data that managers can use in nutrient criteria development, such as in developing reference networks or
    utilizing distribution-based approaches.
Random selection of wetland systems from entire
population within a region.
This design requires minimal prior knowledge of
wetlands within the sample population for stratification.
This design may use more resources (time and money) to
randomly sample wetland classes because more wetlands
may need to be sampled.
System characterization for a class of wetlands is more
statistically robust.
Rare wetlands may be under- represented or absent from
the sampled wetlands.
This design is potentially best for regional
characterization of wetland classes, especially if water
quality conditions are not known.
Targeted selection of wetlands based on problematic
(wetland systems known to have problems) and
reference wetlands.
This design requires prior knowledge of wetlands
within the sample population.
This design utilizes fewer resources because only
targeted systems are sampled.
System characterization for a class of wetlands is less
statistically robust, although characterization of a
targeted wetland may be statistically robust.
This design may miss important wetland systems if
they are not selected for the targeted investigation.
This design is potentially best for site-specific and
watershed-specific criteria development when water
quality conditions for the wetland of interest are
Selection of wetlands based on a known impact.
This design requires knowledge of a specific
impact to be analyzed.
This design may use fewer resources because
only wetlands with known impacts and
associated control systems are sampled.
Characterization of the investigated systems is
statistically robust.
The information gained in this type of
investigation is not transferable to wetland
systems not included in the study.
This design is potentially best for monitoring
restoration or creation of wetlands and systems
that have specific known stressors.
             Table 4: Comparison of Probabilistic, Targeted, and BACI Sampling Designs.
Nutrient Criteria-Wetlands



This chapter provides an overview of candidate variables that could be used to establish nutrient criteria for
wetlands. A more detailed discussion of sampling methods and laboratory analysis with useful references can
be found in the Methods for Evaluating Wetland Condition module series for sampling wetlands4'5 at: http://

A good place to start with selecting candidate variables is by developing a conceptual model of how human
activities affect nutrients and wetlands. These conceptual models may vary from complex to very simple
models, such as relating nitrogen concentrations in sediments and plant biomass or species composition.
Conceptual models establish the detail and scope of the project and the most important variables to select. In
addition, they define the cause-effect relationships that should be documented to determine whether a problem
occurs and what is causing the problem.

In general, for the purposes of numeric nutrient criteria development, it is helpful to develop an understanding
of the relationships among human activities, nutrients and habitat alterations, and attributes of ecosystem
structure and function to establish a simple causal pathway among three basic elements in a conceptual
model. These three basic groups of variables are important to distinguish because we use them differently
in environmental management (Stevenson et al. 2004a). A fourth group of variables is important in order to
account for variation in expected condition of wetlands due to natural variation in landscape setting.

The overview of candidate variables in this chapter follows the outline provided in the conceptual model
in Figure 5.1. Historically, variables in conceptual models have been grouped many ways with a variety of
group names (Paulsen et al. 1991; Stevenson 1998; Stevenson 2004a, b). In this document, three groups and
group names are used to emphasize cause-effect relationships, simplify their presentation and discussion for a
diversity of audiences, and maintain some continuity between their use in the past and their use here. The three
groups are: supporting variables, causal variables, and response variables.

Supporting variables provide information useful in normalizing causal and response variables and categorizing
wetlands. (These are in addition to characteristics used to define wetland classes as described in Chapter 3.)
Causal variables characterize pollution or habitat alterations.

Causal variables are intended to characterize nutrient availability in wetlands and could include nutrient
loading rates and soil nutrient concentrations. Response variables are direct measures or indicators of
ecological properties. Response variables are intended to characterize biotic response and could include
community structure and composition of vegetation and algae. The actual grouping of variables is much less
important than understanding relationships among variables.

It is important to recognize the complex temporal and spatial  structure of wetlands when measuring or
interpreting causal and response variables with respect to nutrient condition. The complex interaction of
climate, geomorphology, soils, and internal interactions has led to a diverse  array of wetland types ranging
from infrequently flooded, isolated depressional wetlands such as seasonal prairie potholes and playa lakes, to
very large, complex systems such as the Everglades and the Okefenokee Swamp. In addition, most wetlands
are complex temporal and spatial mosaics of habitats with distinct structural and functional characteristics
illustrated most visibly by patterns in vegetation structure.
4EPA is developing and revising additional modules as a part of the Methods for Evaluating Wetland Conditions Module Series-Biogeo-
chemical Indicators, Wetland Hydrology, and Nutrient Loading Estimation.
5The references for these modules can be found in the Supplementary References following the References section.
                                        Nutrient Criteria-Wetlands                                       5-1

                                     STRESSORS: CONTAMINANTS a HABITAT ALTERATIONS

                                          RESPONSES; VALUED ECOLOGICAL ATTRIBUTES
                                        Ecosystem Structure
                                         "' ra& Fauna
                 Ecosystem Function
                  Nutrient Retention
                  Hydrologic Regulatio
        This conceptual model illustrates the casual pathway between human activities and valued ecological attributes. It includes
        the role of nutrients in a broader context that includes natural variation among wetlands. The relationship between different
        approaches of grouping variables is illustrated to emphasize the importance of cause/effect relationships.  Here, natural factors
        and human activities regulate the physical, chemical and biological attributes of wetlands.  Some wetland attributes are more
        valued than others and provide the endpoints of assessment and management.  Some physical, chemical, and biological attributes
        are stressors, (i.e., contaminants and habitat alterations) caused by human activities that negatively affect valued ecological
        attributes. The overview of variables in Chapter 5 is organized in three sections: supporting, causal, and response variables.
        Supporting variables are natural landscape level factors that classify expected conditions of wetlands. Causal factors "cause"
        effects in response variables.
         Figure 5.1: Conceptual model of causal pathway between human activities and ecological attributes

     Horizontal zonation is a common feature of wetland ecosystems, and in most wetlands, relatively distinct
     bands of vegetation develop in relation to water depth. Bottomland hardwood forests and prairie pothole
     wetlands provide excellent illustrations of zonation in two very divergent wetland types. However, vegetation
     zones are not static. Seasonal and long-term changes in vegetation structure are a common characteristic of
     most wetland ecosystems. Wetlands may exhibit dramatic shifts in vegetation patterns in response to changes
     in hydrology, with entire wetlands shifting between predominantly emergent vegetation to completely open
     water within only a year or two. Such temporal patterns in fact are important features of many wetlands
     and should be considered in interpreting any causal or response variable. For example, seasonal cycles are
     an essential feature of floodplain forests, which are typically flooded during high spring flows but dry by
     mid to late summer. Longer-term cycles are similarly essential features of prairie pothole wetlands, which
     exhibit striking shifts in vegetation in  response to water level fluctuations over periods of a few years in
     smaller wetlands to decades in larger, more permanent wetlands (van der Valk 2000). Vegetation patterns can
     significantly affect the physical and chemical characteristics of sediments and overlying waters and are likely
     to control major aspects of wetland biogeochemistry and trophic dynamics (Rose and Crumpton 1996).
Nutrient Criteria-Wetlands

The complex temporal and spatial structure of wetlands should influence the selection of variables to measure
and methods for measuring them. Most wetlands are characterized by extremely variable hydrologic and
nutrient loading rates and close coupling of soil and water column processes. As a result, estimates of
nutrient loading may prove more useful than direct measurements of water column nutrient concentrations as
causal variables for establishing the nutrient condition of wetlands. In addition, soil nutrients that integrate a
wetland's variable nutrient history over a period of years may provide the most useful metric against which to
evaluate wetland response.


Supporting variables are not intended to characterize nutrient availability or biotic response but, rather, to
provide information that can be useful in normalizing causal and response variables. Below is a brief overview
of supporting variables that might be useful for categorizing wetlands and for normalizing and interpreting
causal and response variables.

Conductivity (also  called electrical conductance or specific conductance) is an indirect measure of total
dissolved solids. This is due to the ability of water to conduct an electrical current when there are dissolved
ions in solution—water with higher concentrations of dissolved inorganic compounds have higher
conductivity. Conductivity is commonly measured in situ using a handheld probe and conductivity meter
(APHA 1999) or using automated conductivity loggers. Because the conductivity changes with temperature,
the raw measurement should be adjusted to a reference temperature of 25°C. A multiplier of 0.7 is commonly
applied to estimate the total dissolved solids concentration (mg/L) in fresh water when the  conductivity is
measured in units of microsiemens per centimeter (ja,S/cm), although this multiplier varies  with the types of
dissolved ions and  should be adjusted for local chemical conditions.

Conductivity is a useful tool for characterizing wetland inputs and interpreting nutrient condition because of
its sensitivity to changes in these inputs. Rainfall tends to have lower conductivity than surface water, with
ground water often having higher values due to the longer residence time of water in the subsurface. Coastal
and marine waters—as well as water in terminal lakes and wetlands—have even higher conductivity due to
the influence of salinity. Municipal and industrial discharges often have higher conductivity than their intake
waters due to the addition of soluble wastes. Wetland hydrologic inputs can be identified by comparing the
measured input conductivity with the conductivity of potential local  sources.

Soil pH can be important for categorizing wetland soils and interpreting soil nutrient variables. The pH
of wetland soils and water varies over a wide range of values. Many ombrotrophic organic  wetland  soils
(histosols) such as bogs and non-limestone based wetlands are often acidic, and mineral wetland soils are
frequently neutral or alkaline. Flooding a soil results in consumption of electrons and protons. In general,
flooding acidic  soils results in an increase in pH, and flooding alkaline soils decreases pH (Mitsch and
Gosselink 2000). The increase in pH of low pH (acidic) wetland soils is largely due to the reduction of iron and
manganese oxides. However, the initial decrease in pH of alkaline wetland soils is due to rapid decomposition
of soil organic matter and accumulation of CO2. The decrease in pH that generally occurs when alkaline soils
are flooded results  from the buildup of CO2 and carbonic acid. In addition, the pH of alkaline soils is highly
sensitive to changes in the partial pressure of CO2. Carbonates of iron and manganese also can buffer the pH
of soil to neutrality. Soil pH determinations should be made on wet soil samples. Once the  soils are air-dried,
oxidation of various reduced compounds results in a decrease in pH and the values may not represent ambient
                                        Nutrient Criteria-Wetlands                                      5-3

    Soil pH is measured using commercially available combination electrodes on soil slurries. If air dry or moist
    soil is used, a 1:1 soil to water ratio should be used. For details on methodology, the reader is referred to
    Thomas (1996).

    Soil pH can explain the availability and retention capacity of phosphorus. For example, phosphorus
    bioavailability is highest at soil pH near neutral conditions. For mineral soils, phosphorus adsorption capacity
    has been directly linked to extractable iron and aluminum. For details, the reader is referred to Supplementary

    Soil bulk density is the mass of dry solids per unit volume of soil, which includes the volume of solids plus
    air- and water-filled pore space. Bulk density is a useful parameter for expressing the concentration of nutrients
    on a volume basis, rather than mass basis. For example, concentration of nutrients in organic wetland soils can
    be high when expressed on a mass basis (mg/kg or ug/g of dry soil), as compared to mineral wetland soils.
    However, the difference in concentration may not be as high when expressed on a volume (cm3) basis, which is
    calculated as the product of bulk density and nutrient concentration per gram of soil. Expressing soil nutrient
    concentrations on a volume basis is especially relevant to uptake by vegetation since plant roots explore
    a specific volume, not mass, of soil. Expressing nutrients on a volume basis also helps in calculating total
    nutrient storage in a defined soil layer.

    Bulk density is measured by collecting an intact soil core of known volume at specific depths in the soil (Blake
    and Hartge 1986). Cores are oven-dried at 70°C and weighed. Bulk density is calculated as follows:

    Bulk density (dry) (g/cm3) = mass dry weight (grams)/volume (cm3)

    Bulk densities of wetland organic  soils range from 0.1 to 0.5 g/cm3, whereas bulk densities of mineral wetland
    soils range from 0.5 to 1.5 g/cm3. Soil bulk densities are directly related to soil organic matter content, as bulk
    densities decrease with increases in soil organic matter content.

    Soil organic matter can be important for categorizing wetland soils and interpreting soil nutrient variables.
    Wetland soils often are characterized by the accumulation of organic  matter because rates of primary
    production often exceed rates of decomposition. Some wetlands accumulate thick layers of organic matter that,
    over time, form peat soil. Organic matter provides nutrient storage and supply, increases the cation exchange
    capacity of soils, enhances adsorption or deactivation of organic chemicals and trace metals, and improves
    overall soil structure, which results in improved air and water movement. A number of methods are now
    routinely used to estimate soil organic matter content expressed as total organic carbon or loss on ignition
    (APHA 1999; Nelson and Sommers 1996).

    Soil organic matter content represents the soil organic carbon content of soils. Typically, soil organic matter
    content is approximately 1.7 to 1.8 times that of total organic carbon.  The carbon to nitrogen and carbon to
    phosphorus ratios of soils can provide an indication of nutrient availability in soils.

    Wetland hydrologic condition is important for characterizing wetlands and for normalizing many causal and
    response variables. Hydrologic conditions can directly affect the chemical and physical processes governing
    nutrient and suspended solids dynamics within wetlands (Mitsch and Gosselink 2000). Detailed, site-specific
    hydrologic information available is best, but at a minimum, some estimate of water level fluctuation should
    be made. A defining characteristic of wetlands is oxygen deficiency in the soil caused by flooding or soil
    saturation. These conditions influence vegetation dynamics through differential growth and survival of plant
    species and also exert significant control over biogeochemical processes involved in carbon flow and nutrient
5-4                                      Nutrient Criteria-Wetlands

cycling within wetlands. Spatial and temporal patterns in hydrology can create complex patterns in soil and
water column oxygen availability, including alternating aerobic and anaerobic conditions in wetland soils, with
obvious implications for plant response and biogeochemical process dynamics. Water levels in wetlands can
be determined using a staff gauge when surface water is present. A staff gauge measures the depth of surface
flooding relative to a reference point such as the soil surface. Other methods to assess past water levels when
standing water is not present include moss collars, staining, and cypress knee heights. While surface flooding
may be rare or absent in a wetland, high water tables may still cause soil saturation in the rooting zone. In
wetlands where soils are saturated, water level can be measured with a small diameter perforated tube installed
in the soil to a specified depth (Amoozegar and Warrick 1986). Automated water level recorders using floats,
capacitance probes, or pressure transducers are suitable for measuring water levels both above- and below-
ground. The reader is referred to the Supplementary References for details.


Causal variables are intended to characterize  nutrient availability in wetlands. Most wetlands are  characterized
by extremely variable nutrient loading rates and close coupling of soil and water column processes. As a
result, estimates of nutrient loading and measurements of soil nutrients may prove more useful than direct
measurements of water column nutrient concentrations as causal variables for establishing the nutrient
condition of wetlands. Nutrient loading history and soil nutrient measures can integrate a wetland's variable
nutrient history over a period of years and may provide especially useful metrics against which to evaluate
nutrient condition. Wetlands  exhibit a high degree of spatial heterogeneity in chemical composition of soil
layers, and areas impacted by nutrients may exhibit more variability than unimpacted areas of the same
wetland. Thus, sampling protocols should capture this spatial variability. Developing nutrient criteria and
monitoring the success of nutrient management programs involves important considerations for sampling
designed to capture spatial and temporal patterns.

Below is a brief overview of the use of nutrient loading and soil and water column nutrient measures for
estimating nutrient condition of wetlands. Please refer to Supplementary References for a list of references on
both nutrient load estimation and biogeochemical indicators, with a focus on soil and water column nutrient

External nutrient loads to wetlands are determined primarily by surface and subsurface transport from the
contributing landscape, and vary significantly as a function of weather and landscape characteristics such as
soils, topography, and land use. Most wetlands are characterized by extremely variable hydrologic and nutrient
loading rates, which present considerable obstacles to obtaining adequate direct measurement of nutrient
inputs. Adequate measurement of loads may require automated samplers capable of providing flow-weighted
samples when  loading rates are highly variable. In many cases, nonpoint source loads simply may not be
adequately sampled. The more detailed the loading measurements the better, but it is not reasonable to expect
adequate direct measurement of loads for most wetlands. In the absence of sufficient, direct measurements, it
may be possible to estimate nutrient loading using an appropriate loading model or at least to provide a relative
ranking of wetlands based on expected nutrient load. One advantage of loading models is that nutrient loading
can be integrated over the appropriate time scale for characterizing wetland nutrient condition and, in some
cases, historical loading patterns can be reconstructed. Loading models also can provide hydrologic loading
rates to calculate critical supporting variables such as hydroperiod and residence times.

Loading function models are based on empirical or semi-empirical relationships that provide estimates of
pollutant loads on the basis of long-term measurements of flow and contaminant concentration. Generally,
loading function models contain procedures for estimating pollutant load based on empirical relationships
between landscape physiographic characteristics and phenomena that control pollutant export. McElroy et
al., (1976) and Mills (1985) described loading functions employed in screening models developed by the
                                        Nutrient Criteria-Wetlands                                      5-5

    USEPA to facilitate estimation of nutrient loads from point and nonpoint sources. The models contain simple
    empirical expressions that relate the magnitude of nonpoint pollutant load to readily available or measurable
    input parameters such as soils, land use and cover, land management practices, and topography. Preston
    and Brakebill (1999) described a spatial regression model that relates the water quality conditions within a
    watershed to sources of nutrients and to those factors that influence transport of the nutrients. The regression
    model, Spatially-Referenced Regressions on Watersheds (SPARROW), involves a statistical technique that
    utilizes spatially referenced information and data to provide estimates of nutrient load (Smith et al. 1997; Smith
    et al. 2003; http://water.usgs.gov/nawqa/sparrow/).

    In general, the SPARROW methodology was designed to provide statistically based relationships between
    stream water quality and anthropogenic factors such as contaminant sources within the contributing
    watersheds, land surface characteristics that influence the delivery of pollutants to the stream, and in-stream
    contaminant losses via chemical and biological process pathways. The Generalized Watershed Loading
    Functions (GWLF) model (Haith and Shoemaker 1987; Haith et al. 1992) uses daily time steps, and to some
    extent, both can be used to examine seasonal variability and the response to landscape characteristics of
    specific watersheds. The GWLF model was developed to evaluate the point and nonpoint loading of nitrogen
    and phosphorus in urban and rural watersheds. The model enhances assessment of effectiveness of certain
    land use management practices and makes extensive use of readily available watershed data. The GWLF also
    provides an analytical tool to identify and rank critical areas of a watershed and evaluate alternative land
    management programs.

    Process-oriented simulation models attempt to explicitly represent biological, chemical, and physical  processes
    controlling hydrology and pollutant transport. These models are at least partly mechanistic in nature and
    are built from equations that contain directly definable, observable parameters. Examples of process-
    oriented simulation models that have been used to predict watershed hydrology and water quality include the
    Agricultural Nonpoint Source model (AGNPS), the Hydrologic Simulation Program-Fortran (HSPF), and
    the Soil and Water Assessment Tool (SWAT). AGNPS (Young et. al. 1987) is a distributed parameter, event-
    based and continuous simulation model that predicts the behavior of runoff, sediment, nutrients, and pesticide
    transport from watersheds that have agriculture as the primary land use. Because of its simplicity and ease
    of use, AGNPS is probably one of the most widely used hydrologic and water quality models of watershed
    assessment. HSPF (Johansen et al.  1984; Bicknell et al. 1993; Donigian et al. 1995a) is a lumped parameter,
    continuous simulation model developed during the mid-1970s to predict watershed hydrology and water
    quality for both conventional and toxic organic pollutants. HSPF is one of the most comprehensive models
    available for simulating nonpoint source nutrient loading. The capability, strengths, and weaknesses of HSPF
    have been demonstrated by its application to many urban and rural watersheds and basins (e.g., Donigian et al.
    1990; Moore et al. 1992; and Ball et al. 1993). SWAT  (Arnold et al. 1995) is a lumped parameter, continuous
    simulation model  developed by USDA-Agricultural Research Services that provides long-term simulation of
    impact of land management practices on water, sediment, and agricultural chemical yields in large complex
    watersheds. Because of its lumped parameter nature,  coupled with its extensive climatic, soil, and management
    databases, the SWAT model is one of the most widely used hydrologic and water quality models for large
    watersheds and basins, and the model has found widespread application in many modeling studies that involve
    systemic evaluation of impact of agricultural management on water quality.

    These loading models address only gross, external nutrient inputs. It is important to consider the overall
    mass balance for the receiving wetland in developing measures of nutrient loading against which to evaluate
    wetland nutrient condition. This requires some estimate of nutrient export, storage, and transformation. In the
    absence of sufficient, direct measurements from which to calculate nutrient mass balance, it may be possible
    to estimate nutrient mass  balances using an appropriate wetland model.  Strictly empirical, regression models
    can be used to estimate nutrient retention and export in wetlands but these regressions are of little value
    outside the data domain in which they are developed. When developed for a diverse set of systems, the scatter
    in these regressions can be quite large. In contrast to strictly empirical regressions, mass balance models
5-6                                     Nutrient Criteria-Wetlands

incorporate principles of mass conservation. These models integrate external loading to the wetland, nutrient
transformation and retention within the wetland, and nutrient export from the wetland. Mass balance models
allow time varying hydrologic and nutrient inputs and can provide estimates of spatial nutrient distribution
within the wetland. The most difficult problem is developing removal rate equations which adequately
represent nutrient transformation and retention across the range of conditions for which estimates are needed.

Identifying land uses in regions surrounding wetlands is important for characterizing reference condition,
identifying reference wetlands, and providing indicators of nutrient loading rates for criteria development.
Most simply, the percentage of natural area or the percentage of agricultural and urban lands can be used to
characterize  land uses around wetlands.  More detailed quantitative data can be gathered from GIS analysis,
which provides higher resolution identification of land use types such as pastures, row crops, and confined
animal feeding operations for agriculture. Ideally these characterizations should be done for the entire source
shed, including both air and water, in the regions around wetlands. Air-sheds should incorporate potential
atmospheric sources of nutrients, and watersheds should incorporate potential aquatic sources. However, in
practice, land use around wetlands is typically used for defining reference wetlands and also in most nutrient
loading models to characterize groundwater and surface water sources. Land use in buffer zones, one kilometer
zones around wetlands and wetland watersheds (delineated by elevation), has been used to characterize human
activities that could be affecting wetlands (Brooks et al. 2004).

Ammonium is the dominant form of inorganic N in wetland soils, and unlike total soil N (Craft et al. 1995;
Chiang  et al. 2000), soil extractable NH4-N increases in response to N loadings. Enrichment leads to enhanced
cycling  of N between wetland biota (Valiela and Teal 1974; Broome et al. 1975; Chalmers 1979; Shaver et al.
1998), greater activity of denitrifying bacteria (Johnston 1991; Groffman 1994; White and Reddy 1999), and
accelerated organic matter and N accumulation in soil (Reddy et al. 1993; Craft and Richardson 1998). In
most cases, extractable soil N should be measured in the surface soil where roots and biological activity are

Extractable N is  measured by extraction of inorganic (NH4-N) N with 2 M KC1 (Mulvaney 1996). Ten to
20 grams of field moist soil is equilibrated with 100 ml of 2 M KC1 for one hour on a reciprocating shaker,
followed by filtration through Whatman No. 42 filter paper. Ammonium-N in soil extracts is determined
colorimetrically  using the phenate or salicylate method (APHA 1999, Method 350.2; USEPA 1993a).

Extractable P is often a reliable indicator of the P enrichment of soils, and in wetlands,  extractable P is strongly
correlated with surface water P concentration and P enrichment from external sources (Reddy et al. 1995;
1998). Selected methods used to extract P are described below (Kuo 1996). Many soil testing laboratories
perform these analyses on a routine basis. Historically,  these methods have been used to determine nutrient
needs of agronomic crops, but the methods have been used more recently to estimate P impacts in upland and
wetland soils (Sharpley et al. 1992; Nair et al. 1995; Reddy et al. 1995, 1998).

The Mehlich I method is typically used in the Southeast and Mid-Atlantic regions on mineral soils with pH of
< 7.0 (Kuo 1996). The extractant consists of dilute concentrations of strong acids. Many plant nutrients such as
P, K, Ca, Mg, Fe, Zn, and Cu extracted with Mehlich I methods have been calibrated for production of crops
in agricultural ecosystems. This solvent extracts some Fe and Al- bound P, and some Ca-bound P. Soil (dry)
to extractant ratio is set at 1:4 for mineral soils, while wider ratios are used for organic soils.  Soil solutions are
equilibrated for a period of five minutes on a mechanical shaker and then filtered through a Whatman No. 42
filter. Filtered solutions are analyzed for P and other nutrients using standard methods (Method 365.1, USEPA
                                         Nutrient Criteria-Wetlands                                      5-7

    The Bray P-l method has been widely used as an index of available P in soils (Kuo 1996). The combination of
    dilute concentration of strong acid (HC1 at 0.025 M) and ammonium fluoride (NH4F at 0.03 M) is designed to
    easily remove acid extractable soluble P forms such as Ca-bound P, and some Fe and Al-bound P. Soil (dry) to
    extractant ratio is set at 1:7 for mineral soils with wider ratios used for highly organic soils, then shaken for five
    minutes and filtered through a Whatman No.  42 filter. Filtered solutions are analyzed for P and other nutrients
    using the same methods used for the Mehlich I extraction (Method 365.1, USEPA 1993a).

    Bicarbonate Extractable P is a suitable method for calcareous soils. Soil P is extracted from the soil with 0.5
    M NaHCO3 at a nearly constant pH of 8.5 (Kuo 1996). In calcareous, alkaline, or neutral soils containing Ca-
    bound P, this extractant decreases the concentration of Ca in solution by causing precipitation of Ca as CaCO3.
    As a result, P concentration in soil solution increases. Soil (dry) to extraction ratio is set at 1:20 for mineral
    soils and 1:100 for highly organic soils. Soil solutions are equilibrated for a period of 30 minutes on a shaker,
    filtered through a Whatman No. 42 filter paper, and analyzed for P using standard methods (Method 365.1,
    USEPA 1993a).

    Nutrient enrichment leads to enrichment of total soil  P (Craft and Richardson 1993; Reddy et al. 1993;
    Bridgham et al. 2001). In  contrast, soil total N usually does not increase in response to nutrient enrichment
    (Craft et al. 1995; Chiang et al. 2000). Rather, enrichment leads to enhanced cycling of N between wetland
    biota that is reflected in greater N uptake and net primary production (NPP) of wetland vegetation (Valiela
    and Teal 1974; Broome et al. 1975; Chalmers  1979; Shaver et al. 1998),  greater activity of denitrifying bacteria
    (Johnston 1991, Groffman 1994; White and Reddy 1999), and accelerated organic matter and N accumulation
    in soil (Reddy et al. 1993; Craft and Richardson 1998). In most cases, total N and P should be measured in at
    least the surface soil where most roots and biological activity are concentrated.

    Since ammonium N is the dominant form of inorganic nitrogen in saturated wetland soils with very little
    nitrate (NO3) present, total Kjeldahl nitrogen (TKN) can generally be taken as a measure of total N in such
    soils. The difference between TKN and ammonium N provides information on soil organic N. The soil organic
    carbon to soil organic nitrogen ratio can provide an indication of the soil's capacity to mineralize organic N
    and provide ammonium N to vegetation. TKN in soils is determined by converting organic forms of N to
    NH4-N by digestion with  concentrated H2SO4 at temperatures of 300-350°C (Bremner 1996). The NH4-N in
    digested samples is analyzed using colorimetric (e.g., phenate, salicylate) methods (APHA 1999; Mulvaney

    Total P in soils is determined by oxidation of organic forms of P and acid (nitric-perchloric acid) dissolution
    of minerals at temperatures of <300°C (Kuo 1996). Digested solutions are analyzed for P using colorimetric
    methods (e.g., ascorbic acid-molybdate) (APHA 1999; Kuo 1996). Many laboratories may not have access
    to perchloric acid fume-hoods. Alternatively, soil total phosphorus can be determined using the ashing
    method (Anderson 1976). Results obtained from this method are reliable and comparable to total phosphorus
    measurements made using perchloric acid digestion method.

    Nutrient inputs to wetlands are highly variable across space and time, hence, single measurements  of water
    column N and P represent only a "snap-shot" of nutrient condition and may or may not reflect the long-term
    pattern of nutrient inputs  that alter biogeochemical cycles and affect wetland biota. The best use of water
    column N and P concentrations for nutrient criteria development will be based on frequent monitoring of
    nutrient concentrations over time (e.g., weekly or monthly measurements). Of course, in wetlands that are
    seldom flooded, measurements of water column N and P may not be practical or even relevant for assessing
    impacts. Whenever water samples are obtained, it is important that the water depth is recorded because
    nutrient concentration is related to water depth. In the case of tidal estuarine or freshwater wetlands, it is also
    important to record flow and the point in the tidal cycle that the samples were collected.
5-8                                      Nutrient Criteria-Wetlands

Methodologies to monitor N in surface waters are well developed for other ecosystems and can be readily adopted
for wetlands. The most commonly monitored N species are total Kjeldahl nitrogen (TKN), ammonium N, and
nitrate plus nitrite N (APHA 1999). The TKN analysis includes both organic and ammonium N, but does not
include nitrate plus nitrite N. Organic N is determined as the difference between TKN and NH4-N. Forms of N
in surface water are measured by standard methods, including phenol-hypochlorite for ammonium N, cadmium
reduction of nitrate to nitrite for nitrate N, and Kjeldahl digestion of total N to ammonium for analysis of total
N (APHA 1999). Dissolved organic N is primarily used by heterotrophic microbes, whereas plants and various
microorganisms take up inorganic forms of N (ammonium N and nitrate N) to support metabolism and new

Methodologies to monitor P in surface waters are well developed for aquatic ecosystems and can be readily
adopted for wetlands (APHA 1999). The most commonly measured forms of P in surface water are total P,
dissolved inorganic P (i.e., PO4-P), and total dissolved P. To trace the transport and transformations of P  in
wetlands, it might be useful to distinguish four forms of P: (1) dissolved inorganic P (DIP, also referred to as
dissolved reactive P (DRP) or soluble reactive phosphorous (SRP)); (2) dissolved organic P (DOP); (3) particulate
inorganic P (PIP); and, (4) particulate organic P (POP). Dissolved inorganic P (PO4-P) is considered bioavailable
(e.g., available for uptake  and use by microorganisms, algae, and vegetation), whereas organic and particulate
P forms generally must be transformed into inorganic forms before being considered bioavailable. In P limited
wetlands, a significant fraction of DOP can be hydrolyzed by phosphatases and utilized by bacteria, algae, and


Biotic measures that can integrate a wetland's variable nutrient history over a period of months to years may
provide the most useful measures of wetland response to nutrient enrichment. Microorganisms, algae, and
macrophytes respond to nutrient enrichment by: (1) increasing the concentration of nutrients (P, N) in their tissues;
(2) increasing growth and biomass production; and, (3) shifts in species composition. The biotic response to
nutrient enrichment generally occurs in a sequential manner as nutrient uptake occurs first,  followed by  increased
biomass production, followed by a shift in species composition as some species disappear and other species
replace them. Macroinvertebrates respond to nutrient enrichment indirectly as a result of changes in food sources,
habitat structure, and dissolved oxygen. Because of their short life cycle, microorganisms and algae respond more
quickly to nutrient enrichment than macrophytes. However, biotic measures that can integrate a wetland's variable
nutrient history over a period of months to years may provide the most useful measures of wetland response.

Below is a brief overview of the use of macrophytes, algae, and macroinvertebrates to assess nutrient condition of
wetlands. Please refer to the relevant modules in the EPA series "Methods for Evaluating Wetland Condition" for
details on using vegetation (http://www.epa.gov/waterscience/criteria/wetlands/15Indicators.pdf;
http://www.epa.gov/waterscience/criteria/wetlands/16Vegetation.pdf), algae (http://www.epa.gov/waterscience/
criteria/wetlands/11 Algae.pdf); and, macroinvertebrates (http://www.epa.gov/waterscience/criteria/
wetlands/9Invertebrate.pdf) to assess wetland condition, including nutrients.

Wetland macrophytes respond to nutrient enrichment by increasing uptake and storage of N and P (Verhoeven and
Schmitz  1991, Shaver et al. 1998; Chiang et al. 2000). In wetlands where P is the primary limiting nutrient, the
P content of vegetation increases almost immediately (within a few months) in response to nutrient enrichment
(Craft et al. 1995). Increased P uptake by plants is known as "luxury uptake" because P is stored in vacuoles
and used later (Davis 1991). Like P, leaf tissue N may increase in response to N enrichment  (Brinson et al. 1984;
Shaver et al. 1998). However, most N is directly used to support new plant growth so that luxury uptake  of N is not
usually observed (Verhoeven and Schmitz 1991). Tidal marsh grasses, however, do appear to store nitrogen in both
living and dead tissues that can be accessed by living plant tissue. A discussion of conservation and translocation
of N in saltwater tidal marshes can be found in Hopkinson and Schubauer (1980) and Thomas and Christian
                                        Nutrient Criteria-Wetlands                                      5-9

    Nutrient content of macrophyte tissue holds promise as a means to assess nutrient enrichment of wetlands.
    However, several caveats should be kept in mind when using this diagnostic tool (Gerloff 1969; Gerloff and
    Krombholz 1966; EPA 2002c).

            1.  The most appropriate plant parts to sample and analyze should be determined. It is
               generally recognized that the plant or plant parts should be of the same physiological age.

            2.  Samples from the same species should be collected and analyzed. Different species assimilate
               and concentrate nutrients to different levels.

            3.  Tissue nutrient concentrations vary with (leaf) position, plant part, and age. It is important
               to sample and analyze leaves from the same position and age (e.g., third leaf from the terminal
               bud on the plant) to ensure comparability of results from sampling of different wetlands.

            4.  Tissue P may be a more reliable indicator of nutrient condition than N. This is because N is
               used to increase production of aboveground biomass, whereas excess P is stored via luxury

    Another promising macrophyte-based tool is the measurement of nutrient resorption of N and P prior to
    leaf senescence and dieback. Nutrient resorption is an important strategy used by macrophytes to conserve
    nutrients (Hopkinson and Schubauer 1984; Shaver and Melillo 1984). In nutrient-poor environments,
    macrophytes resorb N and P from green leaves prior to senescence, leading to low concentrations of N and P
    in senesced leaves. In nutrient-rich environments, resorption becomes less important so that senesced leaves
    retain much of the N and P that was present when the leaves were green.

    Nitrogen and phosphorus should be measured in green leaves of the same approximate age collected from
    the dominant wetland plant species. Samples also should be collected throughout the wetland to account for
    spatial variability. If an environmental gradient is known or suspected to exist within the wetland, then sites
    along this gradient should be sampled separately. At each sampling location, approximately five green leaves
    are collected from each of the dominant plant species. Leaves are collected from the middle portion of the
    stem, avoiding very young leaves at the top of the stem and very old leaves at the bottom of the stem. At each
    location, leaf samples by species are combined for analysis,  oven-dried at 70°C, and ground.

    Nitrogen is measured by dry combustion using a CHN analyzer. Phosphorus is measured colorimetrically
    after digestion in strong acid (H2SO4-H2O2) (Allen et al. 1986). Many land-grant universities, State agricultural
    testing laboratories, and environmental consulting laboratories perform these analyses. Contact your local U.S.
    Department of Agriculture office or land-grant agricultural extension office for information on laboratories
    that perform plant tissue nutrient analyses.

    Please  see the EPA Module, Vegetation-based Indicators of Wetland Nutrient Enrichment (http://www.epa.gov/
    waterscience/criteria/wetlands/16Indicators.pdf) for a detailed description of indicators derived from N and P
    content of macrophytes.

    Wetland macrophytes also respond to nutrient enrichment by increased net primary production (NPP) and
    growth if other factors such as light are not limiting growth (Chiang et al. 2000). Net primary production
    is the amount of carbon fixed during photosynthesis that is incorporated into new leaves, stems, and roots.
    Most techniques to measure NPP focus on aboveground biomass and discount root production because it is
    difficult to measure, even though root production may account for 50% of NPP. The simplest way to measure
    aboveground biomass is by harvesting all of the standing material (biomass) at the end of the growing season
    (Broome et al. 1986). The harvest method is  useful for measuring NPP of herbaceous emergent vegetation,
    especially in temperate climates where there is a distinct growing season. If root production desired, it can be
5-10                                    Nutrient Criteria-Wetlands

determined by sequentially harvesting roots at monthly intervals during the year (Valiela et. al. 1976).
Enhanced NPP often is reflected by increased height and, sometimes, stem density of herbaceous emergent
vegetation (Broome et. al. 1983). Because increased stem density may reflect other factors like vigorous clonal
growth, it is not recommended as an indicator of nutrient enrichment.

Aboveground biomass of herbaceous vegetation may be determined by end-of-season harvest of aboveground
plant material in small 0.25 m2 quadrats stratified by macrophyte species or inundation zone (Broome et al.
1986). Stem height of individuals of dominant species is measured in each plot. Height of the five to ten tallest
stems in each plot has been shown to be a reliable indicator of NPP (Broome et al. 1986) that saves time as
compared to height measurements of all stems in the plot. Aboveground biomass is clipped at the end of the
growing season, in late summer or fall. Clipped material is separated into live (biomass) versus dead material,
then dried at 70°C to a constant weight. For stem height and biomass sampling, five to ten plots per vegetation
zone are collected. In forested sites, biomass production is defined as the sum of the leaf and fruit fall and
aboveground wood production (Newbould 1967). Please see the EPA module Vegetation-based Indicators
of Wetland Nutrient Enrichment (http://www.epa.gov/waterscience/criteria/wetlands/16Indicators.pdf) for a
detailed description of sampling aboveground biomass in wetlands.

In some cases, measurements of algal N and P can provide a useful complement to vegetation and soil nutrient
analyses that integrate nutrient history over a period of months in the case of vegetation (Craft et al. 1995), to
years in the case of soils (Craft and Richardson 1998; Chiang et al. 2000). Nutrient concentrations in algae can
integrate variation in water column N and P bioavailability over a time scale of weeks, potentially providing
an indication of the recent nutrient status of a wetland (Fong et al. 1990; Stevenson et al. 2001). Caution is
warranted for this method because it is not useful in all wetlands;  for example, in wetlands where surface
inundation occurs intermittently or for short periods of time, where the water surface is severely shaded as in
some forested wetlands, or under other circumstances where unrelated environmental factors exert primary
control over algal growth.

Algae should be sampled by collecting grab samples from different locations in the wetland to account for
spatial variability in the wetland. If an environmental gradient is known or suspected (i.e., decreasing canopy
or impacted land uses) or exists within the wetland as a result of specific source discharges, then sites along
this gradient should be sampled separately. Comparisons among wetlands or locations within a wetland
should be done on a habitat-specific basis (e.g., phytoplankton vs.  periphyton). Samples are processed in the
same manner as wetland plants to determine N and P content. Nitrogen is determined using a CHN analyzer,
whereas P is measured colorimetrically after acid digestion.

Please see the EPA module Using Algae to Assess Environmental Conditions in Wetlands (http://www.epa.
gov/waterscience/criteria/wetlands/1 lAlgae.pdf) for a detailed description of indicators derived from to N and
P content of algae.

The composition of the plant community and the changes that result from human activities  can be used
as sensitive indicators of the biological integrity of wetland ecosystems. In particular, aggressive, fast-
growing species such as cattail (Typha spp.), giant reed (Phragmites communis), reed canarygrass (Phalaris
arundincea), and other clonal species invade and may eventually come to dominate the macrophyte
community. Data collection methods and analyses for using macrophyte community structure and composition
as an indicator of nutrient enrichment and ecosystem integrity for wetlands are described in Vegetation-based
Indicators of Wetland Nutrient Enrichment (http://www.epa.gov/waterscience/criteria/wetlands/16Indicators.
pdf) and Using Vegetation to Assess Environmental Conditions in Wetlands (http://www.epa.gov/waterscience/
criteria/wetlands/lOVegetation.pdf), respectively.
                                        Nutrient Criteria-Wetlands                                    5-11

    Algae can be used as a valuable indicator of biological and ecological condition of wetlands. Structural
    and functional attributes of algae can be measured including diversity, biomass, chemical composition,
    productivity, and other metabolic functions. Species composition of algae, particularly of the diatoms, is
    commonly used as an indicator of biological integrity and physical and chemical conditions of wetlands.
    Discussions of sampling, data analyses, and interpretation are included in Using Algae to Assess
    Environmental  Conditions in Wetlands (http://www.epa.gov/waterscience/criteria/wetlands/HAlgae.pdf).

    Aquatic invertebrates can be used to assess the biological and ecological condition of wetlands. The approach
    for developing an Index of Biological Integrity for wetlands based on aquatic invertebrates is described in
    Developing an Invertebrate Index of Biological Integrity for Wetlands  (http://www.epa.gov/waterscience/

    5.5 SUMMARY

    Candidate variables to use in determining nutrient condition of wetlands and to help identify appropriate
    nutrient criteria for wetlands consist of supporting variables, causal variables, and response variables.
    Supporting variables provide information useful in normalizing causal and response variables and categorizing
    wetlands. Causal variables are  intended to characterize nutrient availability (or assimilation) in wetlands
    and could include nutrient loading rates and soil nutrient concentrations. Response variables are intended to
    characterize biotic response and could include community structure and composition of macrophytes and

    The complex temporal and spatial structure of wetlands will influence the selection of variables to measure
    and methods for measuring them. The information contained in this chapter is a brief summary of suggested
    analyses that can be used to determine wetland condition with respect to nutrient status. The authors  recognize
    that the candidate variables and analytical methods described here will generally be the most useful for
    identifying wetland nutrient condition, while other methods and analyses may be more appropriate in certain
5-12                                     Nutrient Criteria-Wetlands



A database of relevant water quality information can be an invaluable tool to States as they develop nutrient
criteria. In some cases, existing data are available and can provide additional information that is specific to
the region where criteria are to be set. However, little or no data are available for most regions or parameters
and creating a database of newly gathered data is strongly recommended.  In the case of existing data, the
data should be located and their suitability (type and quality and sufficient associated metadata) ascertained.
It is also important to determine how the data were collected to ensure that future monitoring efforts are
compatible with earlier approaches.

Databases operate much like spreadsheet applications but have greater capabilities. Databases store and
manage large quantities of data and allow viewing and exporting of data sorted in a variety of ways, while
spreadsheets analyze and graphically display small quantities of data. Databases can be used to organize
existing information, store newly gathered monitoring data, and manipulate data for water quality criteria
development. Databases can sort data for export into statistical analyses programs, spreadsheets, and graphics
programs. This chapter will discuss the role of databases  in nutrient criteria development and provide a brief
review of existing sources of nutrient-related water quality information for wetlands.


A database is a collection of information related to a particular subject or purpose. Databases are arranged so
that individual values are kept separate, yet can be linked to other values based on some common denominator
(such as association of time or location). Geographic Information Systems (GIS) are geo-referenced relational
databases that have a geographical component (i.e., spatial platform) in the user interface. Spatial platforms
associated with a database allow geographical display of sets of sorted data. GIS platforms such as ArcView™,
Arclnfo™, and Maplnfo™ are  frequently used to integrate spatial data with monitoring data for watershed
analysis. Data stored in simple  tables, relational databases, or geo-reference databases can also be  located,
retrieved, and manipulated using queries. A query allows the user to find and retrieve only the data that meets
user-specified conditions. Queries can also be used to update or delete multiple records simultaneously and to
perform built-in or custom calculations of data. Data in tables can be analyzed and printed in specific layouts
using reports. Data can be analyzed or presented in a specific way in print by creating a report. The most
effective use of these tools requires a certain  amount of training, expertise, and software support,  especially
when using geo-referenced data.

To facilitate data storage, manipulation, and calculations, it is highly recommended that historical and present-
day data be transferred to a relational database (i.e., Access™). Relational databases store data in tables
as sets of rows and columns and are powerful tools for data manipulation  and initial data reduction. They
allow selection of data by specific, multiple criteria and definition and redefinition of linkages among data
components. Data queries can also be exported to GIS, provided that the data is related to some geo-referenced
coordinate system.
                                        Nutrient Criteria-Wetlands                                      6-1

    Potential Data Sources

    EPA Water Quality Data

    EPA has many programs of national scope that focus on collection and analysis of water quality data. The
    following presents information on several of the databases and national programs that may be useful to water
    quality managers as they compile data for criteria development.  STORET STOrage and RETrieval system
    (STORET) is EPA's national database for water quality and biological data.

    The Environmental Monitoring and Assessment Program is an EPA research program designed to develop
    the tools  necessary to monitor and assess the status and trends of national ecological resources (see EMAP
    Research Strategy on the EMAP Web site: http://www.epa.gov/emap). EMAP's goal is to develop the scientific
    understanding for translating environmental monitoring data from multiple spatial and temporal scales into
    assessments of ecological condition and forecasts of future risks to the sustainability of the Nation's natural
    resources. Data from the EMAP program can be downloaded directly from the EMAP Web site (http://www.
    epa.gov/emap/html/data/index.html). The EMAP Data Directory contains information on available data sets,
    including data and metadata (language that describes the nature and content of data).  Current status of the data
    directory, as well as composite data and metadata files, are available on this Web site.

    U.S. Geological Survey (USGS) Water Data
    The USGS has national and distributed databases on water quantity and quality for waterbodies across the
    nation. Much of the data for rivers and streams are available through the National Water Information System
    (NWIS).  These data are organized by State, Hydrologic Unit Codes (HUCs), latitude and longitude, and other
    descriptive attributes. Most water quality chemical analyses are associated with an instantaneous streamflow
    at the time of sampling and can be  linked to continuous streamflow to compute constituent loads or yields. The
    most convenient method of accessing the local databases is through the USGS State representative. Every State
    office can be reached through the USGS home page at: http://www.usgs.gov.

    USGS data from several national water quality programs covering large regions offer highly controlled and
    consistently collected data that may be particularly useful for nutrient criteria analysis. Two programs, the
    Hydrologic Benchmark Network (HBN) and the National Stream Quality Accounting Network (NASQAN),
    include routine monitoring of rivers and streams over the past 30 years. The HBN consists of 63 relatively
    small, minimally disturbed watersheds. HBN data were collected to investigate naturally-induced changes in
    streamflow and water quality and the effects of airborne substances on water quality. The NASQAN program
    consists of 618 larger, more culturally influenced watersheds. NASQAN data provides information for tracking
    water-quality conditions in major U.S. rivers and streams. The watersheds in both networks include a diverse
    set of climatic, physiographic, and cultural characteristics. Data from the networks have been used to describe
    geographic variations in water-quality concentrations, quantify water-quality trends, estimate rates of chemical
    flux from watersheds, and investigate relations of water quality to the natural environment and anthropogenic
    contaminant sources.

    The Water, Energy, and Biogeochemical Budgets (WEBB) program was developed by USGS to study water,
    energy, and biogeochemical processes in a variety of climatic/regional scenarios. Five ecologically diverse
    watersheds, each with an established data history, were chosen.  This program may prove to be a rich data
    source for ecoregions in which the  five watersheds are located. Many publications on the WEBB project are
    available. See the USGS Web site for more details (http://water.usgs.gov/nrp/webb/about.html).
6-2                                     Nutrient Criteria-Wetlands

US Department of Agriculture (USDA) Agricultural Research Service (ARS)
The USDA ARS houses the Natural Resources and Sustainable Agricultural Systems Scientific Directory
(http://hydrolab.arsusda.gov/arssci.html), which has seven national programs to examine the effect of
agriculture on the environment. The program on Water Quality and Management addresses the role of
agriculture in nonpoint source pollution through research on Agricultural Watershed Management and
Landscape Features, Irrigation and Drainage Management Systems, and Water Quality Protection and
Management Systems. Research is conducted across the country and several models and databases have been
developed. Information on research and program contacts is listed on the Web site (http://www.nps.ars.usda.
gov/programs/nr sas. htm).

Forest Service
The Forest Service has designated research sites across the country, many of which are Long Term Ecological
Research (LTER) sites. Many of the data from these experiments are available in the USFS databases located
on the Web site  (http://www.fs.fed.us/research/). Most of the data are forest-related but may be of use for
determining land uses and questions on silviculture runoff.

National Science Foundation (NSF)
The National Science Foundation (NSF) funds projects for the Long Term Ecological Research (LTER)
Network. The Network is a collaboration of over 1,100 researchers investigating a wide range of ecological
topics at 24 different sites nationwide. The LTER research programs are not only an extremely rich data
source, but also a source of data available to anyone through the Network Information System (NIS), the NSF
data source for LTER sites. Data sets from sites are highly comparable due to standardization of methods and

U.S. Army Corps of Engineers (COE)
The U.S. Army  Corps of Engineers (COE) is responsible for many federal wetland jurisdiction issues.
Although a specific network of water quality monitoring data does not exist, specific studies on wetlands by
the COE may provide suitable data. The COE focuses more on water quantity issues than on water quality
issues. As a result, much of the wetland system data collected by the COE does not include nutrient data.
Nonetheless, the COE does have a large water sampling network and supports USGS and EPA monitoring
efforts in many  programs. A list of the water quality programs that the COE actively participates in can be
found at http://www.usace.army.mil/public.html.

U.S. Department of the Interior, Bureau of Reclamation (BuRec)
The Bureau of Reclamation of the U.S.  Department of the Interior manages many irrigation and water supply
reservoirs in the West, some of which may have wetland applicable data available.  These data focus on water
supply information and limited water quality data. However, real time flow data are collected for rivers
supplying water to BuRec, which may be useful if a flow component of criteria development is chosen. These
data can be gathered on a site-specific basis from the BuRec Web site: http://www.usbr.gov.

State Monitoring Programs
Some States may have wetland water quality data as part of a research study, use attainability analysis (UAA),
or to assess mitigation or nutrient related impacts. Most of this data is collected by State natural resources or
environmental protection agencies, or by regional water management authorities. Data collected by State water
quality monitoring programs can be used for nutrient criteria development and may provide pertinent data
sources, although they may be regionally limited. These data should be available from the agencies responsible
for monitoring.
                                        Nutrient Criteria-Wetlands                                     6-3

    Volunteer Monitoring Programs
    State and local agencies may use volunteer data to screen for water quality problems, establish trends in
    waters that would otherwise be unmonitored, and make planning decisions. Volunteers benefit from learning
    more about their local water resources and identifying what conditions or activities might contribute to
    pollution problems. As a result, volunteers frequently work with clubs, environmental groups, and State or
    local governments to address problem areas. The EPA supports volunteer monitoring and local involvement in
    protecting our water resources.

    Academic and Literature Sources
    Most of the data available on water and soil quality in wetlands is the result of research studies conducted
    by academic institutions. Much of the research conducted by the academic community, however, was not
    conducted for the purpose of spatial or long-term biogeochemical characterization of the nation's wetlands;
    instead, water quality information was often collected to characterize the environmental conditions under
    which a particular study or experiment was conducted. Infrequently, spatial studies of limited extent or
    duration were conducted. Data collected from these sources, therefore, may not be sufficiently representative
    of the population of wetlands within an ecoregion. However, this limited data may be the only information
    available and therefore could be useful for identifying reference conditions or determining where to begin a
    more comprehensive survey to support development of nutrient criteria. Academic research data is available
    from researchers and the scientific literature.


    The value of older historical data is a recurrent problem because data quality is often unknown. Knowledge
    of data quality is also problematic for long-term data repositories such as STORET and long-term State
    databases, where objectives, methods, and investigators may have changed many times over the years.
    The most reliable data tend to be those collected by a single agency using the same protocol. Supporting
    documentation should be examined to determine the consistency of sampling and analytical protocols. The
    suitability of data in large, heterogeneous data repositories for establishing nutrient criteria are described
    below. These same factors need to be taken into account when developing a new database such that future
    investigators will have sufficient information necessary to evaluate the quality of the database.

    Geo-referenced data is extremely valuable in that it allows for aggregating and summarizing data according
    to any GIS coverage desired, whether the data was historically related to a particular coverage theme or not.
    However, many studies conducted prior to the availability and accuracy of hand held Global Positioning
    System (GPS) units relied on narrative and less definitive descriptions of location such as proximity to
    transportation corridor, county, or nearest municipal center. This can make comparison of data, depending
    upon desired spatial resolution, difficult. Knowledge of the rationale and methods of site  selection from
    the original investigators may supply valuable information for determining whether inclusion of the site or
    study in the database is appropriate based on potential bias relative to overall wetland data sources. STORET
    and USGS data associated with the National Hydrography Dataset (NHD) are geo-referenced with latitude,
    longitude, and Reach File 3  (RF3) codes (http://nhd.usgs.gov/). In addition, STORET often contains a site
    description to supplement location information. Metadata of this type, when known, is frequently stored within
    large long-term databases.

    Each separate analytical method yields a unique variable. For example, five ways of measuring TP result
    in five unique variables. Data generated using different analytical methods should not be combined in data
    analyses because methods differ in accuracy, precision, and detection limits. Data generated from one
    method may be too limited, making it important to select the most frequently used analytical methods in the
    database. Data that were generated using the same analytical methods may not always be obvious because of
6-4                                      Nutrient Criteria-Wetlands

synonymous names or analytical methods. Consistency in taxonomic conventions and indicator measurements
is likewise important for biological variables and multimetric indices comparisons. Review of recorded data
and analytical methods by knowledgeable personnel is important to ensure that there are no problems with
data sets developed from a particular database.

Data generated by agencies or laboratories with known quality control/quality assurance protocols are most
reliable. Laboratory Quality Control (QC) data (blanks, spikes, replicates, known standards) are infrequently
reported in larger data repositories. Records of general laboratory quality control protocols and specific
quality control procedures associated with specific data sets are valuable in evaluating data quality. However,
premature elimination of lower quality data can be counterproductive because the increase in variance caused
by analytical laboratory error may be negligible compared to natural variability or sampling error, especially
for nutrients and related water quality parameters. However, data of uncertain and undocumented quality
should not be accepted.

Water column nutrient data can be reported in different units (e.g., ppm, mg/L, mmoles). Reporting of nutrient
data from other strata such as soils, litter, and vegetation can further expand the list of reporting units  (e.g.,
mg/kg, g/kg, %, mg/cm3). In many instances, conversion of units is possible; however, in other instances unit
conversion is not possible or is lacking support information for conversion. Consistency in reporting units and
the need to provide conversion tables cannot be overemphasized.

Selecting data from particular agencies with known, consistent sampling and analytical methods and known
quality will reduce variability due to unknown quality problems. Requesting data review for quality assurance
from the collecting agency will reduce uncertainty about data quality.

Long-term records are critically important for establishing trends. Determining if trends exist in the time
series database is also important for characterizing reference conditions for nutrient criteria. Length of time
series data needed for analyzing nutrient data trends is discussed in Chapter 7.

An index period—the time period most appropriate for sampling—for estimating average concentrations
can be established if nutrient and water quality variables were measured through seasonal cycles. The
index period may be the entire year or the summer growing season. The best index period is determined by
considering wetland characteristics for the region, the quality and quantity of data available, and estimates of
temporal variability (if available). Consideration of the data available relative to longer-term oscillations in
environmental conditions (e.g., dry years, wet years) should also be taken into account such that the data is
representative and appropriate. Additional information and considerations for establishing an index period are
discussed in Chapter 7.

Data may have been collected for specific purposes. Data collected for toxicity analyses, effluent limit
determinations, or other pollution problems may not be useful for developing nutrient criteria. Further, data
collected for specific purposes may not be representative of the region or wetland classes of interest. The
investigator should determine if all wetlands or a subset of the wetlands in the database are representative
of the population of wetlands to be characterized. If a sufficient sample of representative wetlands cannot be
found, then a new survey is strongly recommended.
                                         Nutrient Criteria-Wetlands                                      6-5


    New data should be collected when no data presently exist or the data available are not suitable, and should
    be gathered following the sampling design protocols discussed in Chapter 4. New data collection activities
    for developing nutrient criteria should focus on filling in gaps in the database and collecting spatially
    representative regional monitoring data. In many cases, this may mean starting from scratch because no
    data presently exists or the data available are not suitable. Data gathered under new monitoring programs
    should be imported into databases or spreadsheets and, if comparable, merged with existing data for criteria
    development. It is best to archive the data with as much data-unique information (meta-data) as possible. It
    is always possible to aggregate at a later time, but impossible to separate lumped data without having the
    parameter needed to partition the data set. Redundancy may also be a problem but can more easily be avoided
    when common variables or parameters are kept in each database (i.e., dates may be very important). The
    limitations and qualifications of each data set should be known and data 'tagged', if possible, before combining
    them. The following five factors should be considered when collecting new data and before combining new
    data with existing data sets: representativeness, completeness, comparability, accuracy, and precision.

    Sampling program design (when, where, and how you sample) should produce  samples that are representative
    or typical of the regional area being described and the classes of wetlands present. Sampling designs for
    developing nutrient criteria are addressed in Chapter 4. Databases populated by data from the literature or
    historical studies will not likely provide sufficient spatial or class representation of a region. Data interpretation
    should recognize these gaps and be limited until gaps are filled using additional survey information.

    A QA/QC plan should describe how to complete the data set in order to answer questions posed (with a
    statistical test of given power and confidence) and the precautions being taken to ensure that completeness.
    Data collection procedures should document the extent to which these conditions have been met. Incomplete
    data sets may not invalidate the collected data but may reduce the rigor of statistical analyses. Precautions to
    ensure completeness may include collecting extra samples, having back-up equipment in the field, copying
    field notebooks after each trip, and/or maintaining duplicate sets of data in two locations.

    In order to compare data collected under different sampling programs or by different agencies, sampling
    protocols and analytical methods should demonstrate comparable data. The most efficient way to produce
    comparable data is to use sampling designs and analytical methods that are widely used and accepted and
    examined for compatibility with other monitoring programs prior to initiation of a survey. Comparability
    should be assessed for field sample collection, sample preservation, sample preparation and analysis, and
    among laboratories used for sample analyses.

    To assess the accuracy of field instruments and analytical equipment, a standard  (a sample with a known value)
    should be analyzed and the measurement error or bias determined. Internal standards should periodically be
    checked with external standards provided by acknowledged sources. At Federal,  State, and local government
    levels, the National Institute of Standards and Technology (NIST) provides advisory and research services to
    all agencies by developing, producing, and distributing standard reference materials for vegetation, soils,  and
    sediments.  Standards and methods of calibration are typically included with turbidity meters, pH meters DO
    meters, and DO testing kits. The U.S. EPA, USGS, and some private companies provide reference standards or
    QC samples for nutrients.
6-6                                      Nutrient Criteria-Wetlands

The variability in field measurements and analytical methods should be demonstrated and documented to
identify the source and magnitude of variability when possible. EPA QA/QC guidance provides an explanation
and protocols for measuring sampling variability (USEPA 1998c).

For data reduction, it is important to have a clear idea of the analysis that will be performed and a clear
definition of the sample unit for analysis. For example, a sample unit might be defined as "a wetland during
July-August." For each variable measured, a mean value would then be estimated for each wetland during the
July-August index period on record. Analyses are then conducted on the observations (estimated means) for
each sample unit, not with the raw data. Steps recommended for reducing the data include:

        1.  Selecting the long-term time period for analysis;
        2.  Selecting an index period;
        3.  Selecting relevant variables of interest;
        4.  Identifying the quality of analytical methods;
        5.  Identifying the quality of the data recorded; and,
        6.  Estimating values for analysis (mean, median, minimum, maximum) based on the reduction


The validity and usefulness of data depend on the care with which they were collected, analyzed, and
documented. EPA provides guidance on data quality assurance and quality control (USEPA 1998c) to assure
the quality of data. Factors that should be addressed in a QA/QC plan are elaborated below. The QA/QC plan
should state specific goals for each factor and should describe the methods and protocols used to achieve the

        1.  Who will use the data?
        2.  What the project's goals/objectives/questions or issues are?
        3.  What decision(s) will be made from the information obtained?
        4.  How, when, and where project information will be acquired or generated?
        5.  What possible problems may arise and what actions can be taken to mitigate their impact on the
        6.  What type, quantity, and quality of data are specified?
        7.  How "good" those data have to be to support the decision to be made?
        8.  How the data will be analyzed, assessed, and reported?
                                        Nutrient Criteria-Wetlands                                      6-7




Data analysis is critical to nutrient criteria development. Proper analysis and interpretation of data determine
the scientific defensibility and effectiveness of the criteria. Therefore, it is important to evaluate short- and
long-term goals for wetlands of a given class within the region of concern. These goals should be addressed
when analyzing and interpreting nutrient and response data. Specific objectives to be accomplished through
use of nutrient criteria should be identified and revisited regularly to ensure that goals are being met. The
purpose of this chapter is to explore methods for analyzing data that can be used to develop nutrient criteria
consistent with these goals. Included are techniques to evaluate metrics, to examine or compare distributions
of nutrient exposure or response variables, and to examine nutrient exposure-response relationships.

Statistical analyses are used to interpret monitoring data for criteria development. Statistical methods are data-
driven and range from very simple descriptive statistics to more complex statistical analyses. Generally, the
type of statistical analysis used for criteria development is determined by the source, quality, and quantity of
data available.
                            BCG  MODEL:  SNAP  SHOT
Natural structure & function of biotic community maintained

                 Minimal changes in structure & function
                          Evident changes in structure and
                          minimal changes in function
                                                 Moderate changes in structure &
                                                   minimal changes in function
   Major changes in structure &
   moderate changes in function
                       Severe changes in structure & function
                           INCREASING LEVELS OF STRESSORS
        Figure 7.1: Biological condition gradient model describing biotic community condition as levels of
        stressors increase.
                                     Nutrient Criteria-Wetlands


    Wetland systems should be appropriately classified a priori for nutrient criteria development to minimize
    natural background variation (see Chapter 3). This section discusses some of the factors that should be
    considered when classifying wetland systems and in determining the choice of predictor (causal) and response
    variables to include in the analysis.

    Wetland hydrogeomorphic type http://el.erdc.usace.army.mil/wrap/wrap.html may determine the sensitivity
    of wetlands to nutrient inputs, as well as the interaction of nutrients with other driving factors in producing
    an ecological response. Hydrogeomorphic types differ in landscape position, predominant water source,
    and hydrologic exchanges with adjacent water bodies (Brinson 1993). These factors, in turn, influence water
    residence time, hydrologic regime, and disturbance regime. In general, isolated depressional wetlands will
    have greater residence times than fringe wetlands, which, in turn, will have greater residence times than
    riverine wetlands. Systems with long residence times are likely to behave more like lakes than flow-through
    systems and may show a greater response to cumulative loadings. Thus, nutrient loading rates or indicators
    thereof are likely to be a more sensitive predictor of ecological effects for depressional wetlands, while nutrient
    water column or sediment concentrations are likely to be a more sensitive predictor of responses for riverine
    wetlands. Water column concentrations will influence the response of algal communities, while macrophytes
    derive nutrients from both the water column and sediments. Fringe wetlands are likely to be influenced both
    by concentration of nutrients in the adjacent lake or estuary as well as the accumulation of nutrients within
    these systems from groundwater inflow and, in some cases, riverine inputs. The relative influence of these
    two sources will depend on the exchange rate with the adjacent lake, e.g., through seiche activity (Keough et
    al. 1999; Trebitz et al. 2002). In practice, it is difficult to measure loadings from multiple sources including
    groundwater and exchange with adjacent water bodies. If sediment concentrations are  shown to be a good
    indicator of recent loading rates, then sediment concentrations might be the best predictor to use across

    It may be important to control for ancillary factors when teasing out the relationship between nutrients and
    vegetation community response, particularly if those factors interact with nutrients in  eliciting responses.
    For example, riverine and fringe wetlands differ from basin wetlands in the frequency and intensity of
    disturbance from flooding events or ice. Day et. al. (1988) describe a fertility-disturbance gradient model for
    riverine wetlands describing how the relative dominance of plant guilds with different growth forms and life
    history strategies depends on the interactive effects of productivity, fertility, disturbance, and water level.
    In depressional wetlands, the model could be simplified to include only the interaction of fertility with the
    hydrologic regime. Disturbance regimes and water level could be incorporated into analysis of cause-effect
    relationships either as categorical factors or as covariates.

    The selection of assessment and measurement of response attributes for determining ecological response
    to nutrient loadings should depend, in part, on designated uses assigned to wetlands as part of standards
    development. Designated uses  such as recreation (aesthetics and contact) or drinking water are not typically
    assigned to wetlands; thus,  defining nuisance algal blooms in terms of taste or odor problems or aesthetic
    considerations may not be appropriate for wetlands. Guidance  for the definition of aquatic life use is currently
    being refined to describe six stages of impact along a human disturbance gradient, from pristine reference
    condition to heavily degraded sites (Figure 7.1, Stevenson and Hauer 2002; Davies and Jackson 2006). The
    relative abundance of sensitive native taxa is expected to shift with relatively minor impacts, while organism
    condition or functional attributes are relatively robust to altered loadings. However, if maintenance  of
    ecological integrity of sensitive downstream systems is of concern, then it may be important to measure
    some functional attributes related to nutrient retention. Stevenson and Hauer (2002) have suggested a series
    of "resource condition tiers" analogous to those defined for biological condition but related to ecosystem
    functions. Tier 1 requirements are proposed as "Native structure and function of the hydrologic and
7-2                                      Nutrient Criteria-Wetlands

geomorphic regimes and processes are in the natural range of variation in time and space." Thus maintenance
of structure and function of upstream processes should be protective of downstream biological conditions.


Frequency distributions can aid in the setting of criteria by describing central tendency and variability among
wetlands. Approaches to numeric nutrient criteria development based on frequency distributions do not require
specific knowledge of individual wetland condition prior to setting criteria. Criteria are based on and, in a sense,
developed relative to the conditions of the population of wetlands of a given class.

The simplest statistic describing the shape of distributions refers to quartiles, or the 25th and the 75th percentile.
These can be defined as the observation which has 25% of the observations on one side and 75% on the
other side in the case of the first quartile (25th percentile), or vice versa in the case of the third quartile (75th
percentile). In the same manner, the median is the second quartile or the 50th percentile. Graphically, this is
depicted in boxplots as the box length, the lower extreme represents the first quartile, and the upper extreme
represents the third quartile, the area inside the box encompassing 50% of the data.

Distributions of nutrient exposure metrics or response variables can be developed to represent either an
entire population of wetlands or only a subset of those considered to be minimally impacted. In either case, a
population of wetlands should be defined narrowly enough through classification so that the range in attributes
due to natural variability does not equal or exceed the range in attributes related to anthropogenic effects.
The effects of natural variability can be minimized by classifying wetlands by type and/or region. Nutrient
ecoregions define one potential regional classification system (USEPA2000). Alternatively, thresholds in
landscape or watershed attributes defining natural breakpoints in nutrient concentrations can be determined
objectively through procedures such as classification and regression tree (CART) analysis (Robertson et. al.
2001). If a distribution-based approach is used, periodic reviews using empirical data that relate a measured
value to an ecological attribute or  ecosystem function can validate the assumptions of the chosen percentiles.


Indicators characterized as "response" or "condition" metrics should be distinguished from "stressor" or
"causal" indicators, such as nutrient concentrations (Paulsen et al. 1991; USEPA 1998a; Stevenson 2004a).
While both "response" and "causal" indicators could be used in a single multimetric index, it is recommended
that separate multimetric indices be used for "response" and "causal" assessment. Distinguishing between
"response" and "causal" indices can be accomplished utilizing a risk assessment approach with separate
hazard and exposure assessments that are linked to response-stressor relationships (USEPA 1996, 1998a;
Stevenson 1998; Stevenson et al. 2004a, b). A multimetric index that specifically characterizes "responses"
can be used to clarify goals of management (maintenance or restoration of ecological attributes) and to
measure whether goals have been attained with nutrient management strategies. Response-based multimetric
indices can also be used more directly for natural resource damage assessments than multimetric indices with
response and causal variables.

Factors that should be considered in selecting indicators include conceptual relevance (relevance to the
assessment and ecological function), feasibility of implementation (data collection logistics, information
management, quality assurance, cost), response variability (measurement error, seasonal variability,
interannual variability, spatial variability, discriminatory ability), and interpretation and utility (data quality
objectives, assessment thresholds, link to management actions) (Jackson et  al. 2000). Of these factors, cost,
response variability, and ability to meet data quality objectives can be assessed through  quantitative methods.
An  analytical understanding of the factors that affect wetlands the most will also help States develop the most
effective monitoring and assessment strategies.
                                         Nutrient Criteria-Wetlands                                       7-3

    Designated uses such as contact recreation and drinking water may not be applicable to wetlands, hence, it
    may not be readily apparent what the relative significance of changes in different primary producers is for
    organisms at higher trophic levels. Wetland food webs have traditionally been considered to be detritus-based
    (Odum and de la Cruz 1967; Mann 1972; 1988). However, more recent research on wetland food webs utilizing
    stable isotope analysis have identified the importance of phytoplankton, periphyton, or benthic algae as the
    base of the food chain for higher trophic levels (Fry 1984; Kitting et al. 1984; Sullivan and Moncreiff 1990;
    Hamilton et al. 1992; Newell et al. 1995; Keough et al. 1996); in these cases, it would be particularly important
    to monitor shifts in algal producers.

    Empirical relationships can be derived directly between water quality parameters such as total P or
    transparency and wetland biological responses. Unlike lakes or streams, the level of algal biomass
    corresponding to aesthetic problems or ecological degradation in wetlands is not readily defined, so that
    defining a TP-chlorophyll a relationship based on water column measurements is not likely to be useful.
    However, in some wetlands such as coastal Great Lakes, the loss of submerged aquatic vegetation biomass
    and/or diversity with increased eutrophication provides an ecologically significant endpoint (Lougheed et al.
    2001).  Reductions in submerged plant species diversity was associated with increases in turbidity, total P, total
    N, and chlorophyll a, suggesting that a trophic state index incorporating multiple parameters might be a better
    predictor than a single variable such as total P (Carlson 1977).

    Models describing empirical relationships can include linear or nonlinear univariate forms with a single
    response metric, multivariate with multiple response metrics, a series of linked relationships,  and simulation
    models. The simplest forms of linear univariate approaches are correlation and regression analyses; these
    approaches have the advantage that they are simple to perform and transparent to the general  public. When
    assessment thresholds can be determined based on severity of effect or difference from reference conditions
    such that associated exposure criteria can be derived, linear forms should be adequate. In the  case of nonlinear
    relationships, data can generally be transformed to linearize the relationship. However, if it is desired to
    identify the inflection point in a curvilinear relationship as an indicator of rapid ecological change, alternative
    data analysis methods are available, including changepoint analysis (Richardson and Qian 1999) and piecewise
    iterative regression techniques (Wilkinson 1999).

    Multivariate models are useful for relating nutrient exposure metrics to community-level responses. Both
    parametric and nonparametric (nonmetric dimensional scaling or NMDS) ordination procedures can be used to
    define  axes or gradients of variation in community composition based on relative density, relative abundance,
    or simple presence-absence measures (Gauch 1982; Beals 1984; Heikkila 1987; Growns et al.  1992). Ordination
    scores  then can be regressed against nutrient exposure metrics as an indicator of a composite response
    (McCormick et al. 1996). Direct gradient analysis techniques such as canonical correspondence analysis can be
    used to determine which combination of nutrient exposure variables predict a combination of nutrient response
    variables as a first step in deriving multimetric exposure and response variables (Cooper et al. 1999). Indicator
    analysis can be used to determine which subset of species best discriminate between reference sites with low
    nutrient loadings versus potentially impacted sites with high loadings, or weighted averaging  techniques can
    be used to infer nutrient levels from species composition (McCormick et al. 1996; Cooper et al. 1999; Jensen
    et al. 1999). In the latter case, paleoecological records can be examined to infer historic changes in total P
    levels from macrophyte pollen or diatom frustrates, which will be particularly valuable in the absence of sites
    representing reference condition  (Cooper et al.  1999; Jensen et.  al. 1999).

    Some ecohydrological models have been derived that incorporate the effect of multiple stressors (hydrology,
    eutrophication, acidity) on wetland vegetation, thus providing a link between process-based models and
    community level response (see Olde Venterink and Wassen 1997 for review). These models are based on: (1) a
    combination of expert opinion to estimate species sensitivities,  supplemented by multivariate classification of
    vegetation and environmental data to determine boundaries of species guilds; or, (2) field measurements used
    to derive logistic models to quantify dose-response. These approaches could be used to derive wetland nutrient
7-4                                      Nutrient Criteria-Wetlands

criteria for the U.S. provided that models could be calibrated using species and response curves developed
using data for the U.S. Most multiple-stressor models for wetland vegetation have been calibrated using data
from Western Europe (Olde Venterink and Wassen 1997). Latour and colleagues (Latour and Reiling 1993;
Latour et al. 1994) have suggested a mechanism for setting nutrient standards using the occurrence probability
of species along a trophic gradient to extrapolate maximum tolerable concentrations that protect 95% of

A series of linked empirical relationships for wetlands may be most effective for developing nutrient criteria.
Linked empirical relationships may be most useful in cases where integrative exposure measurements such as
sediment nutrient concentrations are more sensitive predictors of shifts in community composition, or algal
P limitation, or other ecological responses (phosphatase enzyme assays; Qian et al. 2003) than are spatially
and temporally heterogeneous water column nutrient concentrations. In these cases, it may be important
to develop one set of relationships between nutrient loading and exposure indicators for a subset of sites at
which intensive monitoring is done, and another set of relationships between nutrient exposure and ecological
response indicators for a larger sample population (Qian et al. 2003).


Changes in nutrient concentrations within or loadings to wetlands often co-occur with other potential stressors
such as changes in hydrologic regime  and sediment loading. In a few cases, researchers have been able to
separate the simple effects of nutrient addition through manipulations of mesocosms (Busnardo et al. 1992;
Gabor et al. 1994; Murkin et al. 1994;  McDougal et al. 1997; Hann and Goldsborough 1997), segments of
natural systems (Richardson and Qian 1999; Thormann and Bayley 1997), or whole wetlands (Spieles and
Mitsch 2000). In other cases, both simple and interactive effects have been examined experimentally, e.g., to
separate effects of hydrologic regime from nutrient loading (Neill 1990a, b; Neill 1992; Bayley et al. 1985).
If nutrient effects are examined by comparing condition of natural wetlands along a loading or concentration
gradient, effects of other driving factors can be minimized by making comparisons among wetlands of similar
hydrogeomorphic type and climatic regime within a well-defined sampling window. In addition, multivariate
techniques for partitioning effects among multiple factors can be used,  such as partial CCA or partial
redundancy analysis (Cooper et al. 1999; Jensen et al. 1999).


Quantitative methods can be used to assess metric cost, evaluation, response variability, and ability to
meet data quality objectives. The most appropriate method varies with respect to the indicator or variable
being considered. In general, statistical techniques are aimed at making conjectures or inferences about
a population's values or relationships between variables in a sample randomly taken from the population
of interest. In these terms, population is defined as all possible values that a certain parameter may take.
For example, in the case of total phosphorus levels present in marsh sediments in nutrient ecoregion VII,
the total population would be determined if all the marshes in that ecoregion were sampled, which would
negate the need for data analysis. Practically, a sample is taken from the population and the characteristics
associated with that sample (mean, standard deviation) are "transferred" to the  entire population. Many of
the basic statistical techniques are designed to quantify the reliability of this transferred estimate by placing
a confidence interval over the sample-derived parameter. More complex forms  of data analysis involve
comparisons of these parameters from different populations (for example, comparison between sites)  or the
establishment of complex data models that are thought to better describe the original population structure
(for example, regression). They are still basic inference techniques that utilize sample characteristics to make
conjectures about the original population.

A basic and typical issue facing any type of sampling design is the number of samples that should be taken
to be confident in the translation from samples to population. The degree of confidence required should
                                        Nutrient Criteria-Wetlands                                      7-5

    be defined as data quality objectives by the end-user and identify the expected statistical rigor for those
    objectives to be met. There are extensive texts on types and manners of sampling schemes; these will not
    be discussed here. This section is geared to determining the minimum data set recommended to work with
    subsequent sections of the data analysis chapter. In interpreting the results of various forms of data analysis,
    an acceptable level  of statistical error is formulated; this is called Type I error, or alpha (a). Type I error can
    be defined as the probability of rejecting the null hypothesis (HO) when this is actually true. In setting the
    Type I error rate, the Type II error rate is also specified.  The Type II error rate, or beta (P), is  defined as failing
    to reject the null hypothesis when it is actually false, i.e., declaring that no significant effect exists when in
    reality this is the case.  In setting the Type I error rate, an acceptable level of risk is recommended; the risk of
    concluding that a significance exists when this is not the case in reality, i.e., the risk of a "false positive"(Type
    I error) or "false negative" (Type  II error). The concepts of Type I and Type II errors are introduced in Chapter
    4 with reference to  sampling design and monitoring, and more fully discussed in Chapter 8 with reference to
    criteria development.

    In experimental or  sampling design, of greater interest is a statistic associated with beta (P), specifically 1 - P,
    which is the power  of a statistical test. Power is the ability of the statistical test to indicate significance based
    on the probability that  it will reject a false null hypothesis. Statistical power depends on the level of acceptable
    statistical significance  (usually expressed as a probability 0.05  - 0.001 (5% -1%) and termed the a level); the
    level of power dictates the probability of "success," or identifying the effect.  Statistical power is a function of
    three factors: effect size, alpha (a), and sample  size, the relationship between  the three factors being relatively

            1. Effect size is defined  as the actual magnitude of the effect of interest. This could  be the difference
               between two means or the actual correlation between the variables. The relationship between the
               effect size  and power is intuitive; if the effect size is large (for example, a large difference between
               means this results in a concomitantly large power.
            2. Alpha is related to power; to achieve a higher level of significance, power decreases if other factors
               are kept constant.
            3. Sample size. Generally, this is the easiest factor to  control. If the  two preceding factors are set,
               increased sample sizes will always result in a greater power.

    As indicated before, the relationship between these three factors is complex and depends on the nature of the
    intended statistical  analysis. An online guide for selecting appropriate statistical procedures is available at:
    http://www.socialresearchmethods.net/. Software packages for performing power analysis have been reviewed
    by Thomas and Krebs  (1997). Online power calculations have been made available by several statistical faculty
    and are available at these Web sites: http://www.yorku.ca/isr/scs/, http://www.surveysystem.com/sscalc.htm,
    http://www.stat.ohio-state.edu/~jch/ssinput.html, and  http://www.stat.uiowa.edu. Additional  Web sites are
    listed in Chapter 4 that emphasize designs for monitoring with statistical rigor.

    Metric response variability can be evaluated by examining the signal to noise ratio along a gradient of nutrient
    concentrations or loading  rates (Reddy et al. 1999). The power of regression analyses can be determined
    using the  power function for a t-test. Optimization of the design, such as the  spacing, number of levels of
    observations, and replication at each level, depend on the purpose of the regression analysis (Neter et al. 1983).

    Multiple correlation analysis can  compound uncertainty and in some instances misidentify correlations due
    to chance as relevant. Appropriate corrections (e.g.,  Bonferroni) should be applied to avoid these errors (Rice

    Multimetric indices are valuable for summarizing and communicating results of environmental  assessments.
    Use  of multimetric  indices is one approach in developing criteria. Furthermore, preservation  of the biotic
7-6                                       Nutrient Criteria-Wetlands

integrity of algal assemblages, as well as fish and macroinvertebrate assemblages, may be an objective
for establishing nutrient criteria. Multimetric indices for stream macroinvertebrates and fish are common
(e.g., Kerans and Karr 1994; Barbour et al. 1999), and multimetric indices with benthic algae have recently
been developed and tested on a relatively limited basis (Kentucky Division of Water 1993; Hill et al. 2000).
Efforts are underway to develop multi-metric indices of biotic integrity for wetlands, and methods modules
are available for characterizing wetland algal, plant, macroinvertebrate, amphibian, and bird communities
(http://www.epa.gov/waterscience/criteria/wetlands/). Methods for multi-metric indices are well developed for
streams and are readily transferable to wetlands. However, higher trophic levels do not often directly respond
to nutrients and therefore may not be as sensitive to relatively small changes in nutrient concentrations as algal
assemblages. It is recommended that relations between biotic integrity of algal or vegetation assemblages
and nutrients be defined and then related to biotic integrity of macroinvertebrate and fish assemblages in a
stepwise, mechanistic fashion. The practitioner should realize, however, that wetlands with a history of high
nutrient loadings have often lost the most sensitive species and in these cases higher trophic level species may
prove to be the best indicators of current nutrient loadings and wetland nutrient condition.

This section provides an overview for developing a multimetric index that will indicate shifts in primary
producers that are associated with trophic status in wetlands. The first step in developing a multimetric
index of trophic status is to select a set of ecological attributes that respond to human changes in nutrient
concentrations or loading. Attributes that respond to an increase in human disturbance are referred to as
metrics. Six to 10 metrics should be selected for the index based on their sensitivity to human activities that
increase nutrient availability (loading and concentrations), their precision, and their transferability among
regions and habitat types. Selected metrics also should respond to the breadth of biological responses to
nutrient conditions (see discussion of metric properties in McCormick and Cairns 1994).

Effects of nutrients on primary producers and effects of primary producers on the biotic integrity of
macroinvertebrates and fish should be characterized to aid in developing nutrient criteria that will protect
designated uses related to aquatic life (e.g., Miltner and Rankin 1998; King and Richardson 2002).

Another approach for characterizing biotic integrity of assemblages as a function of trophic status is to
calculate the deviation in species composition or growth forms at assessed sites from composition in the
reference condition. Similarity or dissimilarity indices can be used for the determining the differences in biotic
integrity of a wetland in comparison to the reference condition. Multivariate similarity or dissimilarity indices
need to be calculated for multivariate attributes such as taxonomic composition (Stevenson 1984; Raschke
1993) as defined by relative abundance of different growth forms or species,  or species presence/absence. One
standard form of these indices is percent community similarity (PSc)  (Whittaker 1952):

        PSc= 2j=i,smin(aj,bi)

Here aj is the percentage of the ith species in sample a, and b| is the percentage of the same ith species in a
subsequent sample,  sample b.

A second common community similarity measurement is based on a distance measurement (which is actually
a dissimilarity measurement, rather than similarity measurement, because the index increases with greater
dissimilarity, Stevenson 1984; Pielou 1984). Euclidean distance (ED) is a standard distance dissimilarity index,
Log-transformation of species relative abundances in these calculations can increase precision of metrics
by reducing variability in the most abundant taxa. However, the practitioner should also be aware that
transformation, while reducing variability, often decreases sensitivity and the ability to distinguish true fine
                                         Nutrient Criteria-Wetlands                                       7-7

    scale changes in community and species composition. Theoretically and empirically, we expect to find that
    multivariate attributes based on taxonomic composition more precisely and sensitively respond to nutrient
    conditions than do univariate attributes, for instance multimetric algal assemblages (see discussions in
    Stevenson and Pan 1999).

    To develop the multimetric index, metrics should be selected and their values normalized to a standard range
    such that they all increase with trophic status. Criteria for selecting metrics can be found in McCormick and
    Cairns (1994) or many other references. Basically, sensitive and precise metrics should be selected for the
    multimetric index and selected metrics should represent a broad range of impacts and, perhaps, designated
    uses. Values can be normalized to a standard range using many techniques. For example, if 10 metrics are used
    and the maximum value of the multimetric index is defined as 100, all 10 metrics should be normalized to the
    range of 10 so that the sum of all metrics would range between 0 and 100. The multimetric index is calculated
    as the sum of all metrics measured in a system.  A high value of this multimetric index of trophic status
    would indicate high impacts of nutrients and should be a robust (certain and transferable) and moderately
    sensitive indicator of nutrient impacts in a stream. A  1-3-5  scaling  technique is commonly used with aquatic
    invertebrates (Barbour et al. 1999; Karr and Chu 1999) and could be used with a  multimetric index of trophic
    status as well. Using the 95th percentile when developing metrics is an approach  that may decrease the
    influence of outliers (Mack 2004).


    When evaluating the relationships between nutrients  and primary producer response within wetland systems,
    it is important to first understand which nutrient is limiting. Once the limiting nutrient is defined, critical
    nutrient concentrations can be specified and nutrient-response relationships developed.

    Defining the Limiting Nutrient
    The  first step in identifying nutrient-producer relationships should be to define the limiting nutrient. Limiting
    nutrients will control biomass and productivity within a system. However, non-limiting nutrients may
    have other impacts, e.g., toxicological effects related to ammonia concentrations  in sediments or effects on
    competitive interactions that determine vegetation community composition (Guesewell et al. 2003). A review
    of fertilization studies indicated that vegetation N:P mass ratios are a good predictor of the nature of nutrient
    limitation in wetlands, with N:P ratios > 16 indicating P limitation at a community level, and N:P ratios < 14
    indicative of N limitation (Koerselman and Meuleman 1996). Guesewell et al.  (2003) found that vegetation
    N:P ratios were a good predictor of community-level  biomass response to fertilization by N or P, but for
    individual species were only predictive of P-limitation and could not distinguish  between N-limitation, co-
    limitation, or no limitation.  Likewise, N, P, and K levels in wet meadow and fen vegetation were found to be
    correlated with estimated supply rates or extractable  fractions in soils (Odle Venterink et al. 2002). A  survey of
    literature values of vegetation and soil total N:P ratios by Bedford  et al., (1999) indicated that many temperate
    North American wetlands are either P-limited or co-limited by N and P, especially those with organic  soils.
    Only marshes have N:P ratios in both soils and plants indicative of N limitation, while soils data suggest that
    most swamps are also N-limited.

    Many experimental procedures are used to determine which nutrient (N, P, or carbon) limits algal growth.
    Algal growth potential (AGP) bioassays are very useful for determining the limiting nutrient (USEPA 1971).
    Yet,  results from such assays usually agree with what would have been predicted from N:P biomass ratios, and
    in some cases N:P ratios in the water. Limiting nutrient-potential biomass relationships  from AGP bottle tests
    are useful in projecting maximum potential biomass in standing or slow-moving  water bodies. However, they
    are not as useful in fast-flowing, and/or gravel or cobble bed environments. Also, the AGP bioassay utilizes a
    single species, which may not be representative of the response of the natural species assemblage.
7-8                                      Nutrient Criteria-Wetlands

Limitation may be detected by other means, such as alkaline-phosphatase activity, to determine if phosphorus
is limiting. Alkaline phosphatase is an extracellular enzyme excreted by some algal species and from roots
in some macrophytes in response to P limitation. This enzyme hydrolyzes phosphate ester bonds, releasing
orthophosphate (PO4) from organic phosphorus compounds (Mullholland et al. 1991). Therefore, the
concentration of alkaline phosphatase in the water can be used to assess the degree of P limitation. Alkaline
phosphatase activity, monitored over time in a wetland, can be used to assess the influence of P loads on the
growth limitation of algae (Richardson and Qian 1999).

There have been no empirical relationships published relating nutrient concentrations or inputs to wetland
chlorophyll a or productivity levels as there have been for streams and lakes. This is likely due to the large
number of factors interacting with nutrients that determine net ecological effects in wetlands. For example,
eutrophication of Great Lakes coastal wetlands and increases in agricultural area in upstream watersheds
have been correlated with decreases  in diversity of submerged aquatic vegetation, yet researchers were unable
to uncouple the effects of nutrients from those of turbidity (Lougheed et al. 2001). Even in experimentally
controlled settings, where it is possible to separate increased suspended solids loadings from nutrient loadings,
effects of nutrients depend heavily on other factors such as periodicity of nutrient additions (pulse vs. press
loadings; Gabor et al. 1994; Murkin et al. 1994; Hann and Goldsborough 1997; McDougal et al. 1997), water
regime (Neill 1990a, b; Thormann and Bayley 1997), food web structure (Goldsborough and Robinson 1996),
and time lags (Neill 1990a, b). It is important in experimental settings to utilize adequate controls for water
additions that may accompany nutrients (Bayley et al. 1985); in empirical comparisons from field data, it
may be difficult if not impossible to separate out these effects.  Day et al.,  (1988) propose a general conceptual
model describing responses of different wetland plant guilds in riverine wetlands based on a combination of
disturbance regime, hydrologic regime, and nutrients. In the latter case, proper classification of sites based
on disturbance and hydrologic regime prior to describing reference condition help to adequately separate out
nutrient-related effects and explain differences in response.

The significance of food web structure in determining nutrient effects does not preclude deriving predictive
nutrient-primary producer relationships or minimize the importance of describing significant impacts.
However, it does highlight the importance of adequately characterizing the trophic structure of wetlands prior
to comparison, especially the number of trophic levels (e.g., presence or absence of planktivorous fish), and
examining interactive effects on multiple classes of primary producers: phytoplankton, epipelon, epiphytic
algae, metaphyton, and macrophytes (Goldsborough and Robinson 1996;  McDougal et al. 1997). In some cases,
addition of nutrients may have little or no effect on some components  such as benthic algae, but can create
significant shifts in primary productivity among others, such as a loss of macrophytes and associated epiphytes
with an increase in inedible filamentous metaphyton and shading of the water column (McDougal et al. 1997).
                                        Nutrient Criteria-Wetlands                                       7-9




This chapter describes recommendations for setting scientifically defensible criteria for nutrients in wetlands
by using data that address causal and biotic response variables. Causal variables (external nutrient loading,
soil extractable P, soil extractable N, total soil N and P, and water column N and P), and biotic response
variables (vegetation N and P, biomass, species composition, and algal N and P) and the supporting variables
(hydrologic condition, conductivity, soil pH, soil bulk density, particle size distribution, and soil organic
matter), as described in Chapter 5 provide an overview of environmental conditions and nutrient status of the
wetland; these parameters are considered critical to nutrient assessment in wetlands. Several recommended
approaches that water quality managers can use to derive numeric criteria in combination with other biological
response variables are presented. These recommended approaches can be used alone, in combination, or
may be modified for use by State water quality managers to derive criteria for wetlands that are scientifically
defensible and protective of the designated use. Criteria developed from multiple lines of evidence using
combined approaches  will provide the greatest scientific defensibility. Recommended approaches for numeric
nutrient criteria development presented here include:

        • the use of reference conditions to characterize natural or minimally impaired wetland systems
          with respect to causal and exposure indicator variables;
        • applying predictive relationships to select nutrient concentrations that will protect wetland
          structure and/or function; and,
        • developing criteria from established nutrient exposure-response relationships (as in the peer-
          reviewed published literature).

The first approach is based on the assumption that maintaining nutrient levels within the range of values
measured for reference systems will maintain the biological integrity of wetlands.  This presumes that a
sufficient number of reference  systems can be identified. The second two approaches are response-based;
hence, the level of nutrients associated with biological impairment should be used to identify criteria. Ideally,
both kinds of information (background variability and exposure-response relationships) will be available for
criteria development. Recommendations are also presented for deriving criteria based on the potential for
effects to downstream receiving waters (i.e., the lake, reservoir, stream, or estuary influenced by wetlands).
States should consider relating these measures to metrics of ecological integrity and periodically assessing
measures to verify assumptions made in criteria development. The chapter concludes with a recommended
process for evaluating proposed criteria, suggestions of how to interpret and apply criteria, considerations for
sampling for comparison to criteria, potential modifications to established criteria, and adoption of criteria into
water quality standards.

The Regional Technical Assistance Group (RTAG) is composed of State and Regional specialists who will
help the Agency and States  establish nutrient criteria for adoption into their water  quality standards. Expert
evaluations are important throughout the criteria development process. The data upon which criteria are based
and the analyses performed to  arrive at criteria should be assessed for veracity and applicability.


The following discussions focus on three general methods that can be used in developing nutrient criteria.
First, identification of reference or control systems for each established wetland type and class should be based
on either best professional judgment or percentile selections of data plotted as frequency distributions. The
second method uses refinement of classification systems, models, and/or examination of system biological
attributes to assess the relationships among nutrients, vegetation or algae, soil, and other variables. Finally, the
third method identifies published nutrient and vegetation, algal, and soil relationships and values that may be
                                         Nutrient Criteria-Wetlands                                      8-1

    used (or modified for use) as criteria. A weight of evidence approach with multiple attributes that combines one
    or more of these three approaches should produce criteria of greater scientific validity.

    One approach to consider in setting criteria is the concept of reference condition. This approach involves using
    relatively undisturbed wetlands as reference systems to serve as examples for the natural or least disturbed
    ecological conditions of a region. These approaches are most useful for estimating reference conditions
    appropriate to the specific designated use for a class of wetlands. Three recommended ways of using reference
    condition to establish criteria are:

             1. Characterize reference systems for each class within a region using best professional judgment
               and use these reference conditions to define criteria.
             2. Identify the 75th to 95th percentile of the frequency distribution for a class of reference wetlands
               as defined in Chapter 3  and use this percentile to define the criteria.
             3. Calculate a 5th to 25th percentile of the frequency distribution of the general population of a class
               of wetlands and use the selected percentile to define the criteria.

    Defining the nutrient condition of wetlands within  classes will allow the manager to identify protective criteria
    and determine which systems may benefit from management action. Criteria that are identified using reference
    condition approaches may require comparisons to similar systems in other States that share the ecoregion
    so that reference condition and developed criteria can be validated. Furthermore, the 95th percentile of the
    reference population and the 5th percentile of the general population are best used to define the  criteria when
    there is great confidence that the group of reference waters truly reflects reference conditions as opposed, for
    example, to best available condition.

    Reference wetlands should be identified for each class of wetland within a State or ecoregion and then
    characterized with respect to external nutrient loading, water column N and P, biotic response variables
    (macrophytes, algae, soils) and supporting environmental conditions. Wetlands classified as reference quality
    should be verified by comparing the data from the reference systems to general population data for each
    wetland class. Reference systems should be minimally disturbed and should have biotic response values that
    reflect this condition.

    Conditions at reference sites may be characterized  using either of two frequency distribution approaches (see
    2 and 3 above). In both approaches, an optimal reference condition value is selected from the distribution of
    an available set of wetland data for a given wetland class. This approach may be of limited value at this time
    because few States currently collect wetland monitoring data. However, as more wetlands are monitored and
    more data become available, this approach may become more viable.

    In the first frequency distribution approach, a percentile (75th to 95th is recommended) is selected from the
    distribution of causal and biotic response variables of reference systems selected a priori based on very specific
    criteria (i.e., highest quality or least impacted wetlands for that wetland class within a region). The values
    for variables at the selected quartile may be used as the basis for nutrient criteria. The selection of a specific
    percentile as the basis for the criterion should be determined by the uses designated for that water.

    If reference wetlands of a given class are rare within a given region or if inadequate information is available to
    assign wetlands with historic nutrient data as "reference" versus "impacted" wetlands,  another approach may
    be appropriate. The second frequency distribution approach involves selecting a percentile of: (1) all wetland
    data in the class (reference and non-reference); or, (2) a random sample distribution of all wetland data within a
    particular class. Due to the random selection process, a lower percentile should be selected because the sample
    distribution is expected to contain some degraded systems. This option is most useful in regions where the
    number of legitimate "natural" reference wetlands  is usually very small, such as in highly developed land use
8-2                                      Nutrient Criteria-Wetlands

areas (e.g., the agricultural lands of the Midwest and the urbanized east or west coasts). EPA's recommendation
in this case is the 5th to 25th percentile depending upon the number of "natural" reference systems available.
If almost all systems are impaired to some extent, then a lower percentile, generally the 5th percentile, is
recommended for selection of reference wetlands.

Both the 75th percentile for the subset of reference systems and the 5th to 25th percentile from a representative
random sample distribution are only recommendations. The actual distribution of the observations should
be the major determinant of the threshold point chosen. For example, a bi-modal distribution of sediment
or water-column nutrients might indicate a natural breakpoint between reference and enriched systems. To
illustrate, Figure 8.1 shows both options and illustrates the presumption that these two alternative methods
should approach a common reference condition along a continuum of data points. In this illustration, the 75th
percentile of the reference data distribution produces an extractable soil P reference condition that corresponds
to the 25th percentile of the random sample distribution.

The choice of a distribution cut-off to define the upper range of reference wetland nutrient levels is analogous
to defining an acceptable level of Type I error, the frequency for rejecting wetlands as members of the
"unimpacted" class when in fact they are part of the reference wetland population (a false designation of
impairment). If a distribution cut-off of 25% is chosen, the rate of falsely designating wetlands as impaired will
be higher than if a distribution cutoff of 5% is chosen; however, the frequency of committing Type II errors
(failing to identify anthropogenically-enriched wetlands) will be lower. As described in Chapter 7, there is a
trade-off between Type I and Type II errors. When additional information is available, it may be possible to
justify a range of values that are representative of least-impaired wetlands that would reduce Type I errors on a
system by system basis.
                                 75th percentile of
                                 reference population is
                                 the starting point for
   Effects thresholds
   can help justify
   criterion value.
             Nutrient data from reference waters (blue) or
             from all waters (gold) similar physical
Paired nutrient and effects data from waters
with similar physical characteristics.
         Figure 8.1: Use of frequency distributions of nutrient concentration for establishing criteria (left
         graphic), and use of effects thresholds with nutrient concentration for establishing criteria (right
                                          Nutrient Criteria-Wetlands

    State water quality managers also may consider analyzing wetlands data based on designated use
    classifications. Using this approach, frequency distributions for specific designated uses, as opposed to
    frequency distributions of reference or general populations, could be examined and criteria proposed based
    on maintenance of high quality systems that are representative of each designated use. For example, one
    criterion could be derived that protects superior quality wetland habitat (SWLH), and a second criterion could
    be identified that maintains good quality wetland habitat (function maintained but some loss of sensitive
    species (Figure 8.2); see Office of Water tiered aquatic life use training module (PDF): (http://www.epa.gov/
    waterscience/biocriteria/modules/wetlOl-05-alus-monitoring.pdf). This recommended approach is designated
    as the Tiered Aquatic Life Use (TALU) and is being developed by the EPA Office of Water in a more detailed
    publication. Using this approach, a criterion range is created and a greater number of wetland systems will
    likely be considered protective of the designated use. In this case, emphasis may be shifted from managing
    wetland systems based on a central tendency toward more pristine systems associated with Tiers I and II. This
    approach also will aid in prioritizing systems for protection and restoration. Subsequent management efforts
    using this approach should focus on improving wetland conditions so that, over time, plots of wetland data
    shift to the left (i.e., improved nutrient condition) of their initial position.

    Two fundamental reasons are commonly considered for using biological attributes in developing nutrient
    criteria. The concepts basically promote the use of biotic responses or biocriteria to nutrient enrichment.
    i.e., both rationales support evaluation of physical and chemical conditions in conjunction with biological
    parameters when establishing water quality criteria. The first reason is that the primary goal of environmental
    assessment and management is to protect and restore ecosystem services and ecological attributes, which are
    often closely related to biological features and functions in ecosystems. Therefore, it is the effects  of nutrients
    on the living components of ecosystems that should become the critical determinant of nutrient criteria, rather
    than the actual nutrient concentrations. The second reason for using biocriteria is that attributes of biological
                                                MAINE TALU
                                Native or natural condition
                                                Minimal loss of species; some
                                                density changes may occur
Some replacement of
sensitive-rare species;
functions fully
Some sensitive species
maintained; altered
distributions; functions
largely maintained
        Tolerant species show
        increasing dominance;
        sensitive species are rare;
        functions altered   Severe alteration of
                            structure and function
                               LOW     Increasing Effects of Stressors    HIGH
            Figure 8.2: Tiered Aquatic Life Use model used in Maine.
           Nutrient Criteria-Wetlands

assemblages usually vary less in space and time than most physical and chemical characteristics measured
in environmental assessments. Thus, fewer mistakes in assessment may occur if biocriteria are employed in
addition to physical and chemical criteria. In those environments where biological attributes change fairly
rapidly, such as in Louisiana's coastal wetland environment where salinity can vary dramatically in response
to wet versus drought years, other techniques will need to be developed. Information on some other techniques
can be found at The Louisiana State University School of the Coast and Environment (http://www.wbi.lsu.edu/
wbi/web-content/index.html) and also in interagency efforts through the Los Angeles Department of Natural
Resources) to assess coastal area ecology. (http://data.lca.gov/Ivan6/app/app_c_ch9.pdf).

Multimetric indices are a special form of indicators of biological condition in which several metrics are used to
summarize and communicate in a single number the state of a complex ecological system. Multimetric indices
for macroinvertebrates and fish are  used successfully to establish biocriteria for aquatic systems in many
States, and several States are developing multimetric indices for wetlands (see http://www.epa.gov/owow Web

Another recommended approach is to identify threshold or non-linear biotic responses to nutrient enrichment.
Some biological attributes respond  linearly with increasing nutrient concentrations, whereas some attributes
change in a non-linear manner. Non-linear changes in metrics indicate thresholds along environmental
gradients where small changes in environmental conditions cause relatively great changes in a biological
attribute. In an example from the Everglades, a specific level of P concentration and loadings was associated
with a dramatic shift in algal composition and loss of the calcareous algal mats typical of this system (Figure
8.3). Overall, metrics or indices that change linearly (typically higher-level community attributes such as
diversity or a multimetric index) provide better variables for establishing biocriteria because they respond
to environmental change along the  entire gradient of human disturbance. However,  metrics that change in a
non-linear manner along environmental gradients are valuable for determining where along the environmental
gradient the physical and chemical  criteria should be set and, correspondingly, how to interpret other biotic
response variables of interest (Stevenson et al. 2004a).

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i i ••*
i 8 10 12
distance Irom Point Source



        Figure 8.3: Percent calcareous algal mat cover in relation to distance from the P source showing the
        loss of the calcareous algal mat in those sites closer to the source (Stevenson et al. 2002).
                                         Nutrient Criteria-Wetlands

    Using Data Published in the Literature
    Values from the published literature may be used to develop nutrient criteria if a strong rationale is presented
    that demonstrates the suitability of these data to the wetland of interest (i.e., the system of interest should
    share the same characteristics with the systems used to derive the published values). Published data, if there
    is enough of it, could be used to develop criteria for: (1) reference condition; (2) predictive (cause and effect)
    relationships between nutrients and biotic response variables; (3) tiered criteria; or, (4) criteria that exhibit
    a threshold response to nutrients. However, published data from similar wetlands should not substitute for
    collection and analysis of data from the wetland or wetlands of interest.

    Considerations for Downstream Receiving Waters
    More stringent nutrient criteria may be appropriate for wetlands that drain into lentic or standing waters.
    For example, it is proposed that 35 ug/L TP concentration and a mean concentration of 8 ug/L chlorophyll
    a constitute the dividing line between eutrophic and mesotrophic lakes (OECD  1982). Natural nutrient
    concentrations in some wetlands may be higher than downstream lakes. In addition, assimilative capacity
    for nutrients without changes in valued attributes may also be higher in wetlands than lakes. Nutrient criteria
    for wetlands draining into lakes may need to be lower than typically would be set if only effects on wetlands
    were considered. This is because EPA's regulations require States to take into consideration the water quality
    standards of downstream waters when designating uses of a water body and adopting appropriate criteria to
    protect those uses.  (See 40 CFR 131.10(b).) Therefore, when adopting nutrient criteria for wetlands draining
    into lakes, States should take into account the protection of the downstream waters of receiving lakes in
    addition to wetlands.

    Following criteria derivation, an expert assessment of the proposed criteria and their applicability to all
    wetlands within the class of interest is encouraged. Criteria should be verified in many cases by comparing
    criteria values for a wetland class within an ecoregion across State boundaries. In fact, development of
    interstate criteria should be an integral part of a State's water quality standards program. In addition, prior
    to recommending any proposed criterion, it is recommended that States take into consideration the water
    quality standards of downstream waters to ensure that their water quality standards provide for attainment and
    maintenance of the water quality standards of downstream waters, (see 40 CFR 131.10(b)). Load estimating
    models, such as those recommended by EPA (USEPA 1999), can assist in this determination (see External
    Nutrient Loading in Chapter 5.3). Water quality managers responsible for downstream receiving waters also
    should be consulted.
8-6                                      Nutrient Criteria-Wetlands


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R-32                                    Nutrient Criteria-Wetlands


ACOE/ACE/COE - Army Corps of Engineers
AGNPS - Agricultural Nonpoint Source Pollution
AGP - Algal Growth Potential
ARS - Agricultural Research Service
BACI - Before/After, Control/Impact
BMP - Best Management Practice
BPJ - Best Professional Judgement
BuRec - Bureau of Reclamation
CART -  Classification and Regression Tree
CCC - Commodity Credit Corporation
CENR - Committee for the Environment and Natural
CGP - Construction General Permit
CHN - Carbon-Hydrogen-Nitrogen
COE - U.S. Army Corps of Engineers
CPGL -  Conservation of Private Grazing Land
CPP - Continuing Planning Process
CREP - Conservation Reserve Enhancement Program
CRP - Conservation Reserve Program
CSFFCP - Central Florida Flood Control Project
CSO - Combined Sewer Overflow
CWA-Clean Water Act
CZARA - Coastal Zone Act Reauthorization
DIP - Dissolved inorganic phosphorus
DO - Dissolved oxygen
DOP - Dissolved organic phosphorus
DRP - Dissolved reactive phosphorus
ECARP - Environmental Conservation Acreage
Reserve Program
ED - Euclidean Distance
EDAS -  Ecological Data Application System
Eh - Redox potential
EMAP - Environmental Monitoring and Assessment
EPA - U.S. Environmental Protection Agency
EQIP - Environmental Quality Incentive Program
FDEP - Florida Department of Environmental
FIP - Forestry Incentive Program
GIS - Geographic Information System
GPS - Geospatial Positioning System
GRTS - Generalized Random Tessellation Stratified
GWLF - Generalized Watershed Loading Function
HBN - Hydrologic Benchmark Network
HEL - Highly erodible land
HGM - Hydrogeomorphic approach
HSPF - Hydrologic Simulation Program - Fortran
HUC - Hydrologic Unit Codes
IBI - Index of Biotic Integrity
ICI - Invertebrate Community Index
LNWR - Loxahachee National Wildlife Refuge
LTER - Long Term Ecological Research
MPCA - Minnesota Pollution Control Agency
N - Nitrogen
NAAQS - National Ambient Air Quality Standard
NASQAN - National Stream Quality Assessment
NAWQA - National Water Quality Assessment
NHD - National Hydrology Dataset
NIS - Network Information System
NIST - National Institute of Standards and
NOAA - National Oceanic  and Atmospheric
NPDES - National Pollution Discharge Elimination
NPP - Net primary production
NRCS - Natural Resources Conservation Service
NSF - National Science Foundation
NWI - National Wetlands Inventory
NWIS - National Water Information Systems
ONRW - Outstanding Natural Resource Waters
P - Phosphorus
PCB - Polychlorinated biphenyls
PCS - Permit Compliance System
PIP - Particulate inorganic phosphorus
POP - Particulate organic phosphorus
PSA - Particle size analysis
PSc - Percent Community Similarity
QA - Quality Assurance
QA/QC - Quality Assurance/Quality Control
QC - Quality Control
REMAP - Regional Environmental Monitoring and
Assessment Program
RF3 - Reach File 3
RTAG - Regional Technical Assistance Group
SCS - Soil Conservation Service
                                      Nutrient Criteria-Wetlands

    SPARROW - Spatially Referenced Regressions on Watersheds
    SRP - Soluble reactive phosphorus
    STORET - Storage and Retrieval System
    SWAT - Soil and Water Assessment Tool
    SWLH - Superior Quality Wetland Habitat
    TALU - Tiered Aquatic Life Use
    TKN - Total Kjeldahl Nitrogen
    TMDL - Total Maximum Daily Load
    TP - Total Phosphorus
    UAA - Use Attainability Analysis
    USDA - United States Department of Agriculture
    USEPA - United States Environmental Protection Agency
    USFWS - United States Fish and Wildlife Service
    USGS - United States Geological Survey
    WCA - Water Conservation Area
    WEBB - Water, Energy, and Biogeochemical Budgets
    WHIP - Wildlife Habitat Incentive Program
    WLA - Wasteload Allocation
    WQBEL - Water Quality Based Effluent Limit
    WQS  - Water Quality Standard
    WRP - Wetlands Reserve Program
A-2                                    Nutrient Criteria-Wetlands


aquatic ecoregion
Level II ecoregions defined by Omernik according to expected similarity in attributes affecting nutrient supply.

(biological criteria) Narrative or numeric expressions that describe the desired biological condition of aquatic
communities inhabiting particular types of waterbodies and serve as an index of aquatic community health.

cluster analysis
An exploratory multivariate statistical technique that groups similar entities in an hierarchical structure.

Elements of State water quality standards, expressed as constituent concentrations, levels, or narrative statements,
representing a quality of water that supports a particular use. When criteria are met, water quality will generally
protect the designated use (40 CFR 131.3(b)).

designated use
Uses defined in water quality standards for each waterbody or segment whether or not the use is being attained

Unconsolidated sediments comprised of both inorganic and dead and decaying particulate organic matter inhabited
by decomposer microorganisms (Wetzel 1983).

ecological unit
Mapped units that are delineated based on similarity in climate, landform, geomorphology,  geology,  soils,
hydrology, potential vegetation, and water.

A region defined by similarity of climate, landform,  soil, potential  natural  vegetation,  hydrology, and  other
ecologically relevant variables.

emergent vegetation
"Erect, rooted herbaceous  angiosperms that may be temporarily to permanently flooded at the base but do not
tolerate prolonged inundation of the  entire plant; e.g., bulrushes (Scirpus spp.), saltmarsh cordgrass" (Cowardin
etal.  1979).

Abundant in nutrients and having high rates of productivity frequently resulting in oxygen depletion below the
surface layer (Wetzel 1983).

The increase of nutrients  in [waterbodies] either naturally or artificially by pollution (Goldman and Home

GIS (Geographical Information Systems)
A computerized information system that  can input, store, manipulate, analyze, and display  geographically
referenced data to support decision-making processes. (NDWP Water Words Dictionary)
                                         Nutrient Criteria-Wetlands                                      A-3

    HGM, hydrogeomorphic
    Land form characterized by a specific origin, geomorphic setting, water source, and hydrodynamic (NDWP Water
    Words Dictionary)

    index of biotic integrity (IBI)
    An integrative expression of the biological condition that is composed of multiple metrics. Similar to economic
    indexes used for expressing the condition of the economy.

    An area of relatively unchannelized upland  between adjacent streams flowing in approximately the  same

    "Includes  wetlands and deepwater habitats with all of the following characteristics: (1)
    situated in a topographic depression or a dammed river channel; (2) lacking trees, persistent emergents, emergent
    mosses  or lichens with greater than 30% areal coverage; and, (3) total area exceeds 8 ha (20 acres). Similar
    wetland and deepwater habitats totaling less than 8 ha are also included in the Lacustrine System if an active
    wave-formed or bedrock shoreline feature makes up all or part of the boundary, or if the water depth in the deepest
    part of the basin exceeds 2 m (6.6 feet) at low water.. .may be tidal or nontidal, but ocean-derived salinity is always
    less than 0.5%" (Cowardin et al.  1979).

    Relatively still- water environment (Goldman and Home 1983).

    The open water of a body of fresh water.

    Region along the shore of a non-flowing body of water.

    Running-water environment (Goldman and Home 1983).

    (Also known as SAV-Submerged Aquatic Vegetation) Larger aquatic plants, as distinct from the microscopic
    plants, including aquatic mosses, liverworts, angiosperms, ferns, and larger algae as well as vascular plants; no
    precise taxonomic meaning (Goldman and Home 1983).
    micrograms per liter, 10-6 grams per liter

    milligrams per liter, 10-3 grams per liter

    mineral soil flats
    Level wetland landform with predominantly mineral soils

    Receiving water inputs from groundwater, and thus higher in salt content (major ions) and pH than ombrotrophic
A-4                                     Nutrient Criteria-Wetlands

Water with salinity of 0.5 to 30%, due to ocean salts.

Molarity, moles of an element as concentration

Type of statistics that relates one or more independent (explanatory) variables with multiple dependent (response)

Trophic status of a waterbody characterized by a small supply of nutrients (low nutrient release from sediments),
low production of organic matter,  low rates  of decomposition, oxidizing hypolimnetic  condition (high DO)
(Wetzel 1983).

"Nontidal wetlands dominated by trees, shrubs, persistent emergents, emergent mosses orlichens, and all such
wetlands that occur in tidal areas where salinity due to ocean-derived salts is below 0.5%. It also includes wetlands
lacking such vegetation, but with all of the following four characteristics: (1) area less than 8 ha (20 acres); (2)
active wave-formed or bedrock shoreline features lacking; (3) water depth in the deepest part of basin less than 2
m at low water; and, (4) salinity due to ocean-derived salts less than 0.5%"  (Cowardin et al. 1979).

"A type of wetland in which organic matter is produced faster than it is decomposed, resulting in the accumulation
of partially decomposed vegetative material called Peat. In some mires peat  never accumulates to the point where
plants lose contact with water moving through mineral soil. Such mires, dominated by grasslike sedges, are called
Fens. In other mires peat becomes so thick that the surface vegetation is insulated from mineral soil. These plants
depend on precipitation for both water and nutrients. Such mires, dominated by acid forming sphagnum moss, are
called Bogs." (NDWP Water Words Dictionary)

Associated aquatic organisms attached or clinging to stems and leaves of rooted plants or other surfaces projecting
above the  bottom of a waterbody (USEPA 1994).

Evergreen shrub bog, found on Atlantic coastal plain.

riverine wetland
A hydrogeomorphic class of wetlands found in floodplains and riparian zones associated with stream or river

slope wetland
A wetland typically formed at a break in slope  where groundwater discharges to the surface. Typically there is no
standing water.

trophic status
Degree of nutrient enrichment of a waterbody.
                                         Nutrient Criteria-Wetlands                                     A-5

    waters of the U.S.
    Waters of the United States is defined at 40 CFR § 230.3(s)) as including:

             a. All waters that are currently used, were used in the past, or may be susceptible to use in interstate or
               foreign commerce, including all waters that are subject to the ebb and flow of the tide;

             b. All interstate waters, including interstate wet lands; and,

             c. All other waters such as interstate lakes, rivers, streams (including intermittent streams), mudflats,
               sandflats, wetlands, sloughs, prairie potholes, wet meadows, playa lakes, or natural ponds the use,
               degradation, or destruction of which would affect or could affect interstate or foreign commerce
               including any such waters:

             1. That are or could be used by interstate or foreign travelers for recreational or other purposes;

             2. From which fish or  shellfish are or could be taken and sold in interstate or foreign commerce; or,

             3. That are used or could be used for industrial purposes by industries in interstate commerce;

             d. All impoundments of waters otherwise defined as waters of the United States under this definition;

             e. Tributaries of waters identified in paragraphs (a) through (d) of this definition;

             f.  The territorial sea; and,

             g. Wetlands adjacent to waters (other than waters that are themselves wetlands) identified in
               paragraphs (a) through (f) of this definition.

    For further information regarding the scope of 'waters of the U.S.'  in light of the U.S. Supreme Court's 2006
    decision in Rapanos v.  United  States,  see "Clean Water Act Jurisdiction Following the U.S. Supreme Court's
    Decision in Rapanos v. United States & Carabell  v.  United States,"  which was jointly  issued by the  U.S.
    Environmental Protection Agency and the Army Corps of Engineers and is available at: http://www.epa.gov/
    owow/wetlands/pdf/RapanosGuidance6507.pdf, http://www.epa.gov/owow/wetlands/.

    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 CFR§ 230.3 (t)/USACE,33 CFR § 328.3 (b)].
A-6                                      Nutrient Criteria-Wetlands



The Everglades (Figure Bl.l) is the largest subtropical wetland in North America and is widely recognized
for its unique ecological character.  It has been affected for more than a century by rapid population growth
in south Florida. Roughly half of the ecosystem has been drained and converted to agricultural and urban
uses. Among other changes, the conversion of 500,000 acres of the northern Everglades to agriculture (the
Everglades Agricultural Area or EAA) and the subsequent diking of the southern rim of Lake Okeechobee
eliminated the normal seasonal flow of water southward from Lake Okeechobee. Furthermore, the construction
of a complex network of internal canals and levees disrupted the natural sheetflow of water through the system
and created a series of impounded wetlands known as "Water Conservation Areas" or WCAs. This conversion
from a hydrologically open to a highly managed wetland occurred gradually, beginning with the excavation of
four major canals during the 1900-1910 period and culminating with the construction of the Central and South
Florida Flood Control Project (CSFFCP) during the 1950s and 60s (Light and Dineen 1994).

The remnant Everglades is managed for multiple and often conflicting uses including water supply, flood
control,  and the hydrologic needs of the natural ecosystem. Water management operations have altered the
quantity, quality, timing, and delivery of flows to the Everglades relative to the pre-disturbance system;
some parts of the system have been damaged by overdrainage, excessive flooding in other areas has stressed
native vegetation communities.  Changes to the seasonal pattern of flooding and drying have influenced
many ecological processes, including changes in the dominant micro- and macro-phytic vegetation, declines
in critical species, and the nesting success of wading bird populations that rely on drawdowns during a
narrow window of time to concentrate fish prey. Canal inputs containing runoff from agricultural and urban
lands contribute roughly 50% of flows to the managed system and have increased loads of nutrients and
contaminants. In particular, phosphorus (P) has been identified as a key limiting nutrient in the Everglades and
increased inputs of this nutrient have been identified as  a significant factor affecting ecological processes and

The primary source of P to  the pre-disturbance Everglades was rainfall, although seasonal flows from Lake
Okeechobee likely contributed significant P to the northern fringe of the wetland. Prior to the implementation
of P control efforts in the late 1990s, canal flows were estimated to contribute more than half of the P load
to the managed Everglades  (SFWMD 1992). Discharge  from the EAA is the main source of water to the
Everglades, with approximately 500,000 acres of farmland draining southward via SFWMD canals, and is the
major source of anthropogenic P. Significant inputs also come from Lake Okeechobee, a naturally mesotrophic
lake that has also been enriched by agricultural runoff.  Several other agricultural and urban catchments
contribute smaller amounts of P via canal  discharges into various parts of the Everglades. However, in general,
canal P concentrations and loads (and associated wetland concentrations) decline from north to south.

The history of P enrichment and associated ecological impacts is not well documented but probably occurred
at a limited scale for much of the last century. Early reports by the South Florida Water Management District
(e.g., Gleason et al. 1975; Swift and Nicholas 1987) showed an expansion of cattail and changes in the
periphyton community in portions of the northern Everglades receiving EAA runoff. The severity and extent
of P impacts were  more fully recognized by 1988 when the Federal Government sued the State of Florida for
allowing P-enriched discharges and associated impacts  to occur in the Everglades. Settlement of this lawsuit
eventually resulted in the enactment of the Everglades Forever Act by the Florida Legislature in 1994, which
required the Florida Department of Environmental Protection (FDEP) to derive a numeric water quality
criterion for P that would "prevent ecological imbalances in natural populations of flora or fauna" in the
                                       Nutrient Criteria-Wetlands                                     B-1

             Appendix Bl. Figure Bl.l: Major hydrologic units of the remnant Florida Everglades (shaded region)
             including (from north to south) the A.R.M. Loxahacthee National Wildlife Refuge (LNWR), Water
             Conservation Area (WCA) 2A, WCA 3A, and Everglades National Park. Shaded lines represent
             the regional canal and levee system that conveys water southward from Lake Okeechobee and the
             Everglades Agricultural Area to the Everglades and urban areas along the coast.
Nutrient Criteria-Wetlands

Everglades. These legal and legislative events provided the basis for numerous research and monitoring efforts
designed to better understand the effects of P enrichment and to determine levels of enrichment that produced
undesirable ecosystem changes.

Research and monitoring were  initiated by the State of Florida (the Florida Department of Environmental
Protection and the South Florida Water Management District) and other university research groups (e.g., Duke
University, Florida International University, University of Florida) to better understand ecological responses to
anthropogenic P inputs and to identify a P concentration or range of concentrations that result in unacceptable
degradation of the Everglades ecosystem.  This case study reviews research and monitoring conducted by the
State to derive a P criterion for the Everglades. This criterion was proposed by the FDEP in 2001  and approved
in 2003. This process is divided into three parts:

        1.  Define the reference (i.e., historical) conditions for P and the oligotrophic ecology of the
        2.  Determine the types of ecological impacts caused by P enrichment; and,
        3.  Identify wetland P concentrations that produce these impacts, and determine a criterion that
           will protect the resource from those impacts.


Several sources of information were used to characterize reference conditions across the Everglades. Sampling
in minimally impacted locations (i.e., reference sites) believed to best reflect historical conditions provided
the quantitative basis for establishing reference conditions with respect to P concentrations and associated
ecological conditions. Where possible, this characterization was augmented by historical evidence. Written
accounts of surveys conducted  during the  1800s and early 1900s provided useful qualitative data  on past
ecological conditions. Early scientific literature contained substantial information on large-scale  vegetation
patterns (e.g., Davis 1943; Loveless 1959).  Paleoecological assessments, including the dating and analysis of
soil cores with respect to nutrient content and preserved materials  such as pollen provided, further information
(e.g., Cooper and Goman 2001, Willard et  al. 2001).

Predisturbance Everglades exhibited significant spatial and temporal variation, and, while its conversion to a
smaller, more managed wetland resulted in the loss of some of this heterogeneity, the legacy of past variations
in hydrology, chemistry, and biology remain in many areas.  Legislation mandating the development of a P
criterion stipulated that natural variation in P concentrations and ecological conditions within the remnant
ecosystem be considered. This  required that sampling efforts encompass the expected range of background
variability in the remnant ecosystem. To ensure that spatial variation in P conditions were considered,
sampling was conducted in all four major hydrologic units: The Loxahatchee National Wildlife Refuge
(LNWR), WCA-2A, WCA-3A,  and Everglades National Park (see Figure Bl.l).

Nutrient inputs to the Everglades were historically derived primarily from atmospheric deposition (rainfall and
dry fallout), which is typically low in P. Historical loading rates have been estimated from annual atmospheric
P inputs in south Florida and reconstructions of P accumulation in Everglades soils and probably  averaged
less than 0.1 g P m2 yl (SFWMD 1992). Atmospheric inputs of P were augmented by inflows from Lake
Okeechobee, which was connected by surface-water flows to the northern Everglades during periods of high
water (Parker et al.  1955). While inflows from this historically eutrophic lake were undoubtedly enriched in
P compared with the Everglades, the influence of these inputs were likely limited to wetlands along the lake's
southern fringe (Snyder and Davidson 1994) as is demonstrated by the limited extent of pond apple and other
vegetation that require more nutrients for growth than the sawgrass (Cladium jamaicense) that dominates most
of the Everglades.
                                        Nutrient Criteria-Wetlands                                      B-3

    Interior areas of the Everglades generally retain the oligotrophic characteristics of the predrainage ecosystem
    and, thus, provide the best contemporary information on historical P concentrations. Water chemistry data
    were available for several interior locations that had been sampled by the State for many years. Median
    water-column TP concentrations at these stations ranged between 4 and 10 ug I/1, with lowest concentrations
    occurring in southern areas that have been least affected by anthropogenic P loads (Figure B1.2). Phosphorus
    concentrations >10 ug L ' were measured periodically at many of these sites. Isolated high P concentrations
    at reference stations were attributed to P released as a result of oxidation of exposed soils, increased fire
    frequency during droughts, and difficulties in collecting water samples that are not contaminated by flocculent
    wetland sediments when water depths are low. Data from reference sites may represent an upper estimate of
    historical TP concentrations in the Everglades since several stations are located in areas that either have been
    overdrained, a condition which promotes soil oxidation and P release, or so heavily exposed to canal inflows
    (e.g., WCA 2A) that some P inputs have likely intruded even into interior areas. However, in the absence of
    reliable historical data these values were deemed as best available for defining reference condition.

    Extensive soil mapping projects across interior portions of the central and northern Everglades indicate a
    reference range for soil TP in the surface 0-10 cm of soil of between 200 and 500 mg kg"1 on a mass  basis
    (DeBusk et al. 1994; Reddy et al. 1994a, Newman et al. 1997; Richardson et al.  1997a, Newman et al. 1998).
    Fewer data are available from ENP, but available evidence indicates background concentrations of < 400 mg
    kg-1 (Doren et al.  1997).  Soil P content also varies volumetrically as a function of changing soil bulk density.
    The typical bulk density of flooded Everglades peat soils is approximately 0.08 g cm3, whereas soils that have
    been subjected to extended dry out and oxidation can have bulk densities greater than 0.2 g cm3 (Newman
    et al. 1998). Increases in volumetric nutrient concentrations resulting from increased bulk density can have
    a stimulatory effect on plant growth even in the absence of external P inputs (see Chapter 2).  Following
    correction for the varying bulk densities in the peat soils of the Everglades, a historical TP concentration of
    <40 ug cm3 may be applicable for most regions (DeBusk et al. 1994; Reddy et al. 1994a, Newman et al. 1997;
    Newman et al. 1998; Reddy et al. 1998). In the LNWR, most of the interior area has soil TP < 20 ug TP cm3
    (Newman etal.  1997).

Water-column TP (ug/L)
0 I
O O t



Hydrologlc Unit


            Appendix Bl. figure B1.2: Box plots showing surface-water P concentrations at long-term monitoring
            stations in each major hydrologic unit that illustrate the minimally impacted (i.e., reference) condition
            of the Everglades with respect to P. The top, mid-line, and bottom of each box represents the 75th,
            50th (median), and 25th percentile of data, respectively; the error bars represent the 90th and 10th
            percentiles; open circles are data outside the 90th percentile; the dashed line is the analytical limit for
            TP (4 ug I/1).
Nutrient Criteria-Wetlands

The Everglades is perhaps the most intensively studied wetland in the world and, therefore, the ecological
attributes that defined the predisturbance structure and function of this ecosystem are well understood
compared with most wetlands. Clearly, not all of the valued ecological attributes of this or any other wetland
are affected directly by P enrichment. Thus, in order to define the reference condition of the ecosystem with
respect to the role of P, this assessment focused on those processes and communities that are most sensitive
to P enrichment.  Based on available information and preliminary scoping studies, five ecological features
were selected as biotic response variables. These features included three indicators of ecosystem structure,
one indicator of ecosystem function, and one indicator of landscape change. Structural indicators included the
periphyton community, dominant macrophyte populations, and the benthic macroinvertebrate community.
Diel fluctuations  in water column DO provided an important indicator of shifts in aquatic metabolism. The
landscape indicator of change was the loss of open-water slough-wet prairie habitats—areas of high natural
diversity and productivity.

Aquatic vegetation and other submerged surfaces in the oligotrophic Everglades interior are covered with
periphyton, a community of algae, bacteria and other microorganisms. Periphyton accounts for a significant
portion of primary productivity in sloughs and wet prairies (Wood and Maynard 1974; Browder et al. 1982;
McCormick et al. 1998),  and floating and attached periphyton mats provide an important habitat and food
source for invertebrates and small fish (Browder et al.  1994; Rader 1994). These mats store large amounts
of P (approaching 1 kg TP m-2 in some locations) and, thus, may play a critical role in maintaining low P
concentrations in reference areas (McCormick et al. 1998; McCormick and Scinto 1999). Periphyton biomass
and productivity  peak towards the end of the wet season (August through October) and reach a minimum
during the colder months of the dry season (January through March). Periphyton biomass in open-water
habitats can exceed 1 kg  m-2 during the wet season (Wood and Maynard 1974; Browder et al.1982; McCormick
et al. 1998) when floating mats can become so dense as to cover the entire water surface. Aerobic conditions in
slough-wet prairie habitats is maintained by the high productivity of this community and the capacity of dense
algal mats to trap oxygen released during photosynthesis (McCormick and Laing 2003).

Two types of periphyton communities occur in reference areas of the Everglades. Mineral-rich waters, such
as those found WCA 2A  and Taylor Slough (ENP), support a periphyton assemblage dominated by a few
species of calcium-precipitating cyanobacteria and diatoms, while the soft-water interior of LNWR contain a
characteristic assemblage of desmid green algae and diatoms. Waters across much of the southern Everglades
(WCA-3A and portions of ENP) tend to be intermediate with respect to mineral content and contain some taxa
from both assemblages.

The chemical composition of periphyton in the oligotrophic Everglades is indicative of severe P limitation.
Periphyton samples from reference areas of major hydrologic units within the Everglades are characterized
by an extremely low P content (generally <0.05%) and extremely high N:P ratios (generally >60:1 w:w). This
observational evidence for P limitation is supported by experimental fertilization studies that have shown
that: 1) periphyton responds more strongly to P enrichment than to enrichment with other commonly limiting
nutrients such as nitrogen (Scheldt et al.  1989; Vymazal et al.  1994); and, 2) periphyton changes in response
to experimental P enrichment mimic those that occur along field nutrient gradients (McCormick and O'Dell
1996). Thus, it is well-established that periphyton is strongly P-limited in reference areas of the Everglades.

Interior Everglades habitats exhibit characteristic diel fluctuations in water-column dissolved oxygen (DO),
although aerobic  conditions are generally maintained throughout much or all of the diel cycle (Belanger et
al. 1989; McCormick et  al. 1997; McCormick and Laing 2003). High daytime concentrations in open-water
habitats (i.e., sloughs, wet prairies) are a product of photosynthesis by periphyton and other submerged
vegetation. These habitats may serve as oxygen sources for adjacent sawgrass stands, where submerged
                                        Nutrient Criteria-Wetlands                                     B-5

    productivity is low (Belanger et al.  1989). Oxygen concentrations decline rapidly during the night due to
    periphyton and sediment microbial respiration and generally fall below the 5 mg L ' standard for Class III
    Florida waters (Criterion 17-302.560(21), F.A.C.). However, these diurnal excursions are characteristic of
    reference areas throughout the Everglades (McCormick et al. 1997) and are not considered a violation of the
    Class III standard (Nearhoof 1992). In fact, a site specific criterion for DO has been adopted by the State; a
    PDF copy of the technical support document (Weaver 2004) can be found at: http://www.dep.state.fl.us/water/

    The vegetation communities characteristic of the pristine Everglades are dominated by species adapted to
    low P, seasonal patterns of wetting and drying, and periodic natural disturbances such as fire, drought, and
    occasional freezes (Duever et al. 1994; Davis 1943; Steward and Ornes 1983; Parker 1974). Major aquatic
    vegetation habitats in oligotrophic areas include sawgrass wetlands, wet prairies, and sloughs (Loveless 1959;
    Gunderson 1994). The spatial arrangement of these habitats is dynamic and controlled by environmental
    factors such as fire, water depth, nutrient availability, and local topography (Loveless  1959).

    Sawgrass  (Cladium jamaicense) is the dominant macrophyte in the Everglades, and stands of this species
    compromise approximately 65 to 70% of the total vegetation cover of the Everglades (Loveless 1959). Wet
    prairies include a collection of low-stature, graminoid communities occurring on both peat and marl soils
    (Gunderson 1994). Dominant macrophyte  taxa in these habitats include Rhynchospora, Panicum, and
    Eleocharis (Loveless 1959; Craighead 1971). Sloughs are deeper water habitats that remain wet most or all of
    the year and are characterized by floating macrophytes such as fragrant white water lily (Nymphaea odorata),
    floating hearts (Nymphoides Aquaticum),  and spatterdock (Nuphar advena) (Loveless 1959; Gunderson 1994).
    Submerged aquatic plants,  primarily bladderworts (Utricularia foliosa and U. purpurea in particular), also can
    be abundant in these habitats and, in the case of U. purpurea, provide a substrate for the formation of dense
    periphyton mats.

    Several studies have concluded that macrophyte communities in the Everglades are P-limited.  Sawgrass is
    adapted to the low-P conditions indicative of the pristine Everglades (Steward and Ornes 1975b, Steward
    and Ornes 1983). During field and greenhouse manipulations, sawgrass responded to P enrichment either by
    increasing the rate of growth or P uptake (Steward and Ornes 1975a, Steward and Ornes 1983; Craft et al. 1995;
    Miao et al. 1997; Daoust and Childers 1999). Furthermore, additions of N alone had no effect on  sawgrass
    or cattail growth under low-P conditions (Steward and Ornes 1983; Craft et al. 1995). Recent experimental
    evidence in the Everglades National Park (Daoust and Childers 1999) has shown that other native vegetation
    associations such as wet prairie communities are also limited by P.

    Historically, cattail (Typha spp.) was one of several minor macrophyte species native to the Everglades (Davis
    1943; Loveless 1959). In particular, cattail is believed to have been associated largely with areas of disturbance
    such as alligator holes and recent burns (Davis 1994). Analyses of Everglades peat deposits reveal no evidence
    of cattail peat, although the presence of cattail pollen indicates its presence historically in some areas (Gleason
    and Stone 1994; Davis et al. 1994; Bartow et al.  1996). Findings such as these confirm the historical presence
    of cattail in the Everglades but provide no  evidence for the existence of dense cattail stands covering large
    areas (Wood and Tanner 1990)  as now occurs in the northern Everglades. In contrast,  sawgrass and water lily
    peats have been major freshwater Everglades soils for approximately 4,000 years (McDowell et al.  1969).

    Aquatic invertebrates (e.g., insects, snails, and crayfish) represent a key intermediate position in energy flow
    through the Everglades food web as these taxa are the direct consumers of primary production and, in turn,
    are consumed by vertebrate predators. Invertebrates occupy several functional niches within the  Everglades
    food web;  however, most taxa are direct consumers of periphyton and/or plant detritus (e.g., Rader and
    Richardson 1994; McCormick et al.  2004).  Rader (1994) sampled both periphyton and macrophyte habitats in
B-6                                     Nutrient Criteria-Wetlands

this same area and, based on the proportional abundance of different functional groups, suggested that grazer
(periphyton) and detrital (plant) pathways contributed equally to energy flow in low-nutrient areas of the

The macroinvertebrate fauna of the Everglades is fairly diverse (approximately 200 taxa identified) and is
dominated by Diptera (49 taxa), Coleoptera (48 taxa), Gastropoda (17 taxa), Odonata (14 taxa), and Oligochaeta
(11 taxa) (Rader 1999). Most studies have focused on a few conspicuous species (e.g., crayfish and apple
snails) considered to be of special importance to vertebrate predators, and relatively little is known about
the distribution and environmental tolerances of most taxa. An assemblage of benthic microinvertebrates
(meiofauna) dominated by Copepoda and Cladocera is also present in the Everglades (Loftus et al.  1986), but
even less is known about the distribution and ecology of these organisms.

Invertebrates are not distributed evenly among Everglades habitats but, instead, tend to be concentrated in
periphyton-rich habitats such as sloughs. In an early study, Reark (1961) noted that invertebrate densities
in ENP were higher in periphyton habitats compared with sawgrass stands. Rader (1994) reported similar
findings in the northern Everglades and  found mean annual invertebrate densities to  be more than six-fold
higher in sloughs than in sawgrass stands. Invertebrate assemblages in sloughs were  more species-rich
and contained  considerably higher densities of most dominant invertebrate groups. Functionally, slough
invertebrate assemblages contained similar densities of periphyton grazers and detritivores compared with a
detritivore-dominated assemblage in sawgrass stands. Higher invertebrate  densities in  sloughs were attributed
primarily to abundant growths of periphyton and submerged vegetation, which provide oxygen and a source of
high-quality food.

A targeted design (see Chapter 4) was used to quantify changes in key ecological attributes in response to
P enrichment.  Discharges of canal waters through fixed water-control structures are  the primary source of
anthropogenic P for the Everglades and produce P gradients that extend several kilometers into the wetland
in several locations. These gradients have existed for several decades and provided the  clearest example of
the long-term ecological impacts associated with P  enrichment. Monitoring was conducted along gradients in
different parts of the Everglades to assess ecological responses to P enrichment. Fixed  sampling stations were
located along the full extent of each gradient to document ecological conditions associated with increasing
levels of P enrichment. Intensive monitoring was performed along gradients in two northern Everglades
wetlands,  WCA 2A and the LNWR. WCA 2A is a mineral-rich, slightly basic peatland  and contains the most
pronounced and well studied P gradient  in the Everglades, whereas LNWR is a soft-water, slightly acidic
peatland. These two wetlands represent the most extreme natural water chemistry conditions in the Everglades
and support distinct periphyton assemblages and macrophyte populations while sharing dominant species such
as sawgrass and water lily. Less intensive sampling along gradients in other parts of  the Everglades (WCA 3A
and ENP) to confirm that P relationships were consistent across the wetland.

Chemical  and biological conditions were measured at each sampling station along the two intensively sampled
gradients. Repeated sampling, sometimes over several years, was performed to ensure  that temporal variation
in each metric was considered in the final data analysis. Monthly surface-water sampling and less frequent
soil sampling were performed to quantify P gradients in each area. Diel DO regimes, periphyton, and benthic
macroinvertebrates were sampled quarterly when surface water was present.  Macrophyte sampling included
ground-based methods to document shifts in species composition and remote sensing to determine changes
in landscape patterns.  The hydrology of each site was characterized to determine whether P gradients were
confounded with hydrologic gradients, which can also exert a strong influence on ecological patterns.

Numerous field experiments have been conducted to quantify ecological responses to P enrichment and to
better understand how interactions between P enrichment and other factors such as hydrology may affect
these responses. The design of these experiments varied in complexity with respect to size and dosing regimen
                                        Nutrient Criteria-Wetlands                                     B-7

    depending on the specific objective of each study and has included enclosed fertilizer plots (e.g., Craft et al.
    1995), semi-permeable mesocosms receiving periodic P additions to achieve fixed loading rates in the form of
    periodic additions (e.g., McCormick and O'Dell 1996), flumes receiving semi-continuous enrichment at a fixed
    rate (Pan et al. 2000), and flumes receiving flow-adjusted dosing to achieve constant inflow concentrations
    (Childers et al. 2002). These experiments were useful in establishing the causal nature of responses to P
    enrichment documented along the P gradients described above.

    Strong gradients in P concentrations were documented downstream of canal discharges into most Everglades
    wetlands (Figure B1.3). Inflow TP concentrations in from 1996-1999 have averaged as high as 100 jo,g L ' as
    compared with reference and pre-disturbance concentrations < 10 jo,g L ' . The degree and spatial extent of
    P enrichment varies among areas depending on the source and magnitude of inflows. The most extensive
    enrichment has occurred in the northern Everglades near EAA inflows, while southern areas (e.g., ENP) have
    been relatively less affected. The most extensive enrichment has occurred in WCA-2A, which, unlike other
    areas, receives most of its water from canal discharges.  Soil TP was strongly correlated with surface-water
    concentrations and exceeded 1500 mg kg' at the most enriched locations as compared with concentrations   <
    500 mg kg"1 in reference areas. In general, this enrichment effect is limited to the surface 30 cm of soil depth
    (Reddyetal.  1998).

    Ecological Responses to Phosphorous Enrichment

    Periphyton responses to P enrichment include changes in productivity, biomass, and species composition.
    Periphyton rapidly accumulates P from the water (McCormick et al. 2001, Noe et al. 2003), and,  thus, a
    strong relationship between P concentrations in the water and periphyton is maintained along the P gradients
                         120 -
                                     2      4      6      8      10      12
                                           Distance from canal (km)
            Appendix Bl. Figure B1.3: Mean water-column TP concentrations (1996-1999) at long-term
            monitoring stations downstream of canal discharges in two northern Everglades wetlands, WCA 2A
            (circles connected by solid line) and LNWR (squares connected by dashed line). Error bars are + 1 SE.
Nutrient Criteria-Wetlands

(Grimshaw et al. 1993; McCormick et al. 1996). In fact, increases in periphyton P may provide one of the
earliest signals of P enrichment (e.g., Gaiser et al. 2004). Rapid increases in periphyton photo synthetic
activity and growth rates occur in response to P enrichment (e.g., Swift and Nicholas 1987; McCormick et al.
1996; McCormick et al. 2001). All of these responses are consistent with the P-limited nature of Everglades

Paradoxically, these physiological responses are associated with sharply lower periphyton biomass in
P-enriched areas due to the loss of the abundant community of calcareous cyanobacteria and diatoms that
is indicative of mineral-rich reference areas. This community is replaced by a eutrophic community of
filamentous cyanobacteria, filamentous green algae, and diatoms in areas having even slightly elevated
P concentrations. For example, McCormick and O'Dell (1996) found that the calcareous assemblage that
existed at low water-column P concentrations (TP = 5 to 7 (o,g I/1) was replaced by a filamentous green algal
assemblage at moderately elevated concentrations (TP = 10 to 28 jo,g I/1) and by eutrophic cyanobacteria and
diatoms species at even higher concentrations (TP = 42 to  134 jo,g I/1). These results are representative of those
documented by other investigators (e.g., Swift and Nicholas 1987; Pan et al. 2000). Taxonomic changes in
response to controlled P enrichment in field experiments have been shown to be similar to those documented
along field enrichment gradients (Figure B1.4), thereby providing causal evidence that changes in the
periphyton assemblage were largely a product of P enrichment (McCormick and O'Dell 1996; Pan et al. 2000).

Phosphorus enrichment causes a shift in the balance between autotrophy and heterotrophy in the water
column as a result of contrasting effects on periphyton productivity and microbial respiration. Rates of aquatic
primary productivity (P) and respiration (R) are approximately balanced (P:R ratio = 1) across the diel cycle
in minimally impacted sloughs throughout the Everglades (Belanger et al.  1989; McCormick et al.  1997). In
contrast, respiration rates exceed productivity by  a considerable margin (P:R ratio « 1) at enriched locations.
This change is related primarily to a large reduction in areal periphyton productivity as a result of shading
by dense stands of cattail (Typha domingensis) that form a nearly continuous cover in the most enriched
areas (McCormick and Laing, 2003). Increased cattail production also stimulates microbial respiration (e.g.,
sediment oxygen demand) (e.g., Belanger et al. 1989) due to an increase in the quantity and decomposability
of macrophyte litter.

The shift towards dominance of heterotrophic processes with P enrichment, in turn, affects dissolved oxygen
(DO) concentrations in enriched areas. For example, DO concentrations at an enriched site in WCA 2A rarely
exceeded 2 mg L ' compared with concentrations as high as 12 mg L ' at reference locations (McCormick et
al. 1997). Depressed water-column DO concentrations have subsequently been documented in enriched areas
of WCA 2A and the LNWR (Figure B1.5) and confirmed in experimental P-enrichment studies (McCormick
and Laing 2003). Declines in DO along field P gradients were steepest within a range of water-column TP
concentrations roughly between 10 and 30 jo,g I/1. Lower DO in enriched areas are associated with other
changes including an increase in anaerobic microbial processes and a shift in invertebrate species composition
toward species tolerant of low DO, described later in this study.

Nutrient  enrichment initially stimulates the growth of existing vegetation as evidenced by increased plant P
content, photosynthesis, and biomass production, as it does for periphyton. Persistent enrichment eventually
produces a shift in vegetation composition toward species better adapted to rapid growth and expansion under
conditions of high P availability. Two major shifts in Everglades plant communities have been documented
along P gradients, including: (1) the replacement of sawgrass stands by cattail; and, (2) the replacement of
slough-wet prairie habitat by cattail.

Sawgrass populations in the Everglades have life-history characteristics indicative of plants adapted to low-
nutrient environments (Davis 1989; Davis 1994; Miao and Sklar 1998). Sawgrass responses to P enrichment
                                        Nutrient Criteria-Wetlands                                      B-9

             Appendix Bl. figure B1.4: Changes in percent biomass (as biovolume) of major algal groups in
             field enclosures dosed weekly with different P loads (left panel) and along a P enrichment gradient
             downstream of canal discharges (right panel) in WCA 2A. From McCormick and O'Dell (1996).
inimum DO (mg/L) Frequency <1 mg/L
:> ro -£t oioooro ^01 o
Mean DO (mg/L) M
o s s a i e
8 r2 = 0.712
p < 0.001
°/ °
°i$^ ;„ - °o

r^ 0.615
p < 0.001
^g%Jw* rfb

r2 = 0.692
0 p < 0.001
o°8° * o
*» 0"
0 o
o° o °
o o

o r2 = 0.314
00 P = °-OM
0° °

include an increase in tissue P, plant biomass, P storage, annual leaf production and turnover rates, and seed
production (e.g., Davis 1989; Craft and Richardson 1997; Miao and Sklar 1998). Cattail is characterized by
a high growth rate, a short life cycle, high reproductive output, and other traits that confer a competitive
advantage under enriched conditions (Davis  1989; Davis 1994; Goslee and Richardson 1997; Miao and Sklar

Measurements and controlled enrichment experiments have shown that cattail growth rates exceed those
of sawgrass under enriched conditions (Davis 1989; Newman et al. 1996; Miao and DeBusk 1999). The
replacement of sawgrass by cattail in P enriched areas may be facilitated by disturbances such as flooding or
severe fires that weaken or kill sawgrass plants and create openings. Consequently, sawgrass distributional
patterns were not as clearly related to P gradients as were other ecological indicators of enrichment.

Sloughs and wet prairies appear to be particularly sensitive to replacement by cattail under P-enriched
conditions, possibly due to the sparser vegetation cover in these habitats. The process of slough enrichment
and replacement by cattail as shown in satellite imagery is supported by ground-based sampling methods
(McCormick et al.  1999) that documented changes in slough vegetation and encroachment of these habitats
by cattail in areas where soil TP concentrations averaged between 400 and 600 mg kg"1 and water-column TP
in recent years averaged > 10 jo,g I/1. Eleocharis declined in response to  increased soil P, and Nymphaea was
stimulated by enrichment and was dominant in slightly enriched sloughs. Increased occurrence of cattail in
sloughs was associated with a decline in Nymphaea, probably as a result of increased shading of the water
surface. These findings are consistent with those of Vaithiyanathan et al., (1995) who documented a decline in
slough habitats along this same nutrient gradient and the loss of sensitive taxa such as Eleocharis at locations
where soil TP exceeded 700 mg kg~l. As discussed by McCormick et al., (2002), loss of these open-water areas
is a sensitive landscape indicator of P enrichment (Figure B1.6).

Macroinvertebrates are the most widely used biological indicator of water quality impacts, and several changes
that occur in this community along P enrichment gradients in the Everglades are similar to those documented
in response to eutrophication in other aquatic ecosystems. Several studies have documented an overall increase
in macroinvertebrate abundance with increasing P enrichment (Rader and Richardson 1994; Trexler and
Turner et al. 1999; McCormick et al. 2004). However, differences in sampling methodology have apparently
produced conflicting results with respect to changes in species richness  and diversity. For example, Rader and
Richardson (1994) documented an increase in both macroinvertebrate species richness and diversity with P
enrichment in open-water (i.e., low emergent macrophyte cover) habitats and concluded that enrichment had
not impacted this community. McCormick et al., (2004), however, using a landscape approach that involved
habitat-weighted sampling, found little change in either species richness or diversity in response to enrichment.
This latter study accounted for the decline in the cover of habitats such as sloughs and wet prairies, which
contain the most diverse and abundant macroinvertebrate communities  (Rader 1994). McCormick et al.,
(2004) also documented a pronounced shift in community composition with increasing P enrichment as taxa
characteristic of the oligotrophic interior of the wetland are replaced by  common pollution-tolerant taxa of
oligochaetes and chironomids. These changes were indicative of habitat degradation as determined using biotic
indices derived by the Florida DEP to assess stream condition based on  macroinvertebrate composition (results
available at http://www.epa.gov/owow/wetlands/bawwg/case/fl2.html).

As for many other P-induced biological changes, the greatest change in the macroinvertebrate community
occurred in response to relatively small increases in P concentration. Along field enrichment gradients,
community shifts were associated with increases in water-column TP above approximately 10 jo,g I/1
(McCormick et al.  2004). Similarly, Qian et al., (2004) documented several shifts in community structure and
function in response to long-term experimental dosing at average concentrations of approximately 10-15 jo,g
                                        Nutrient Criteria-Wetlands                                     B-11

    The FDEP was charged with reviewing and analyzing available P and ecological data collected throughout
    the Everglades to establish a numeric P criterion. A brief summary of this process is provided here, and more
    detailed information can be found in Payne et al., (2000, 2001a,b; available at http://www.dep.state.fl.us/water/
    wqssp/everglades/pctsd.htm; and, 2002, 2003, available at http://www.sfwmd.gov/sfer/previous_ecr.html).

    The narrative nutrient standard for Class III Florida waters such as the Everglades states that "in no case shall
    nutrient concentrations of a body of water be altered so as to cause an imbalance in natural populations of
    aquatic flora or fauna." The FDEP approach to detecting violations of this standard with respect to surface-
    water P concentrations in the Everglades was to test for statistically significant departures in ecological
    conditions from those at reference sites (i.e., interior sampling locations with background P concentrations).
    Biological and chemical data collected along anthropogenic P gradients throughout the Everglades were
    analyzed to determine P concentrations associated with such departures.  Results showed that sampling sites
    with average  (geometric mean) surface-water TP concentrations significantly greater than 10 ppb consistently
    exhibited significant departures in ecological condition from that of reference sites. A key finding supporting
    this concentration as the standard was the fact that multiple changes in each of the major indicator groups—
    periphyton, dissolved oxygen, macrophytes, and macroinvertebrates—all occurred at or near this same
    concentration (e.g., Payne et al. 2001).

    Data from field and laboratory experiments conducted by various research groups provided valuable
    supporting information for understanding responses to P enrichment. While such experiments were not
    used directly to derive the P criterion, they established cause-effect relationships between P enrichment
    and ecological change that supported correlative relationships documented along field P gradients. For
    example, McCormick and O'Dell (1996) and Pan et al., (2000) showed that major shifts in periphyton species
    composition documented along field P  gradients matched those elicited by controlled P dosing in field
    enrichment experiments.  McCormick and Laing (2003)  confirmed that controlled P enrichment produced
    declines in water-column DO similar to those measured  along the gradients. Macroinvertebrate community
    changes were documented experimentally, Qian et al., (2004).

    While the criterion established a  surface-water concentration of 10 jo,g L ' TP as protective of native flora
    and fauna, the methodology used to measure compliance with the criterion needed to normalize background
    fluctuations in concentration. Additional analyses of P data collected over several years at reference sites was
    used to set both a longer-term average concentration and a shorter-term maximum concentration for each
    site. Based on these analyses, the FDEP concluded that annual maximum concentrations at a given sampling
    location should not exceed 15 jo,g  L ' TP over the long-term, while five-year average concentrations should not
    exceed 10 jo,g L ' TP. These limits would be applied to reference areas to  ensure no further degradation and
    to areas already impacted by P enrichment to gauge the rate and extent of recovery in response to a suite of P
    control measures, including agricultural BMPs and the construction of treatment wetlands to remove P from
    surface runoff prior to being discharged into the Everglades. Additional information on Florida's progress
    in assessing and implementing the adopted standard can be found on the  South Florida Water Management
    District Web  site: www.sfwmd.gov.
B-12                                     Nutrient Criteria-Wetlands




 4       6       8      10      12

Distance from canal (km)

    Appendix Bl. figure B1.6: Changes in the percentage of open-water (i.e., sloughs, wet prairies, or

    other opening caused by natural disturbance or airboats) cover at 94 locations along a P enrichment

    gradient in WCA 2A as determined using aerial
                                   Nutrient Criteria-Wetlands



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L-4                                     Nutrient Criteria-Wetlands

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                                        Nutrient Criteria-Wetlands                                      L-5