Biological Monitoring

In 1995, the West Virginia Department of Environmental Protection (DEP) established the Watershed Assessment Program to assess and report on the water quality of the State's streams and lakes. Following a period of restructuring within the agency, the program's name was changed to the Watershed Assessment Branch (WAB). The WAB is comprised of a staff of biologists and environmental scientists who measure and assess the chemical and physical properties of water, assess habitat conditions, and collect biological samples in the form of benthic macroinvertebrates and fish from streams and lakes throughout the state. As of 2013, the WAB has collected about 8,100 benthic macroinvertebrate samples and 414 fish community samples from many streams in the state. Since 1998, the WAB has used the benthic macroinvertebrate data to measure the biological health of West Virginia streams. Fish community sampling in streams has been part of the WAB assessment procedure since 2006.

Water Sampling Habitat Assessment

Benthic Macroinvertebrates
Benthic macroinvertebrates (also known as "benthos") are small animals living among stones, logs, sediments and aquatic plants on the bottom of streams, rivers and lakes. They are large enough to see with the naked eye (macro) and have no backbone (invertebrate). Insects comprise the largest diversity of the these organisms and include mayflies, stoneflies, caddisflies, beetles, midges, crane flies, dragonflies and others. Non-insect members of the benthic macroinvertebrate community are snails, clams, aquatic worms and crayfish. To date, the WAB has collected and identified 538 different kinds (mostly Genus level identifications) of benthos from 6,202 stations on 5,530 different streams throughout West Virginia. The WAB has found the diversity of some benthos to be especially high, including mayflies (49 genera) and stoneflies (43 genera). Some benthic genera are fairly common throughout the state, while others are limited in their distribution to just a few areas. The most commonly encountered benthos is the midge Polypedilum (Family Chironomidae), found at 81% of collection stations. The caddisfly Cheumatopsyche (Family Hydropsychidae) and mayfly Baetis (Family Baetidae) have been found at 77% and 72% of the stations, respectively. The net-winged midge Blepharicera (Family Blephariceridae) is not common, found at only 1.2% of all collection stations. This sensitive organism is restricted to the steeper streams in the higher mountains of the state, where oxygen rich water cascades over waterfalls and tumbles through small rapids and riffles.

For more information about benthic macroinvertebrates, click on a link below.

WAB Benthic Macroinvertebrate Data Quality Reporting Tool
Guide to Aquatic Invertebrates (West Virginia Save Our Streams)
Benthic Macroinvertebrate Images (West Virginia Save Our Streams)

Benthic Macroinvertebrate and Fish Topics
Habitats Adaptations Importance in Food Webs Biological Indicators Collecting Bethic Macroinvertebrates
Using to Assess Biological Conditions WVSCI Scoring Process WVSCI Example Fish

Caddisfly Family Hydropsychidae Mayfly Family Baetidae Midge Family Blephariceridae

Benthic macroinvertebrates live in a wide variety of habitats and can be found from the smallest headwater streams down to the largest rivers. In West Virginia, benthic macroinvertebrates are especially diverse, having adapted to the complex array of streams that dissect the mountains and tumble through the narrow valleys. In general, benthic organisms are most diverse in the fast flowing riffle and run areas of streams. Compared to pools and glides, riffles and runs are shallower and have higher stream gradients and faster water velocities. They are composed of rough materials, such as large gravel, cobbles and small boulders that create turbulence and oxygenate the water, while providing a stable habitat for benthos to live. Many insect benthos spend the majority of their lives (anywhere from 1 month to 4 years, depending on the species) in the water and only emerge as adults for a few hours (or up to several days) to reproduce and complete their life cycle. Movement of benthos larvae includes swimming, crawling around on the stream bottom and drifting with stream currents. After emerging, adult aquatic insects can fly to new stream locations during their winged terrestrial stage.
Bee Run - Small headwater stream - benthos habitat Gandy Creek - Mountain trout stream - benthos habitat Adult Stonefly

Benthic macroinvertebrates have special adaptations that allow them to live in the stream environment. These adaptations may be unique physical features or even specialized behaviors. For example, many species have flattened bodies that allow them to hide between boulders and cobbles, thus limiting the stress of fast-moving water and allowing them to avoid some predators. Others have sharp claws, suction cups, or other grasping mechanisms on their bodies to prevent them from being swept away in the swift-flowing current. Black fly larvae (Family Simuliidae) have a silk gland and posterior hooks that are used to anchor them in place while they face upstream actively capturing food with their fan-like mouth parts. Net-building caddisfly larvae (Family Hydropsychidae) construct funnel-shaped nets, attach them to stable substrate materials, and then periodically harvest the food caught in the mesh. Some stonefly species will avoid high water temperatures by burrowing deep into the stream bottom substrate where they will diapause near the water table. One of the most important adaptations for benthos is the ability to extract oxygen from the water in which they live. Many aquatic insects accomplish this with external gills that remove oxygen from the surrounding water, while others rely upon cutaneous respiration to obtain oxygen.
Mayfly Family Heptageniidae - flattened body Black Fly Family Simuliidae - silk glands and hooks

Benthos are often classified based on their feeding strategy. The stonefly Pteronarcys (Family Pteronarcyidae) is in the shredder functional feeding group. These large stoneflies primarily consume decaying leaves that have fallen into the stream from trees along the forested riparian corridor. The water penny beetle (Family Psephenidae) is a scraper and can be observed grazing on algae that covers rocks and logs. The mayfly Family Baetidae is classified as a collector-gatherer, primarily feeding on fine pieces of organic material on the stream bottom. The most recognized predator in the aquatic insect world is the larval form of the dobsonfly, more commonly known as the hellgrammite (Family Corydalidae). Hellgrammites are active predators, crawling around on the large cobble and boulders in the swifter habitats of streams where they search for other benthic macroinvertebrates to eat.
Beetle Family Psephenidae - grazing on algae Hellgrammite searching for prey

Importance in Food Webs
Benthic macroinvertebrates are extremely important in aquatic food webs. In most streams, the energy stored by plants is available for consumption by benthos either in the form of leaves that fall into the water or in the form of alagae that grows on the stream bottom. The energy derived by eating leaves and algae is then transferred from benthos to other life forms in and around the stream such as fish, frogs, salamanders, snakes, birds, and even fishermen. In another sense, benthos are important for recreational purposes, as thousands of trout anglers strive to "match the hatch" of many species, particularly those of mayflies.
Brook Trout - preys on benthic macroinvertebrates Mud Salamander - preys on benthic macroinvertebrates

Biological Indicators
Benthic macroinvertebrates provide reliable and comprehensive information on water and habitat quality and have been used as biological indicators in many parts of the world for nearly 100 years. In some cases, it may be difficult to identify pollutants and stressors in streams with chemistry data alone, which only
provides information pertinent to the precise time of sampling. Even the presence of fish may not be indicative of the status of a stream because fish can swim away to avoid polluted water or unfavorable habitat conditions and then return when conditions improve. Many benthic macroinvertebrates are not as mobile as fish and cannot move to avoid pollution. Therefore, the community of benthos living in a stream may indicate the water quality conditions of the past. Additionally, benthos are excellent tools for assessing water quality because they are extremely diverse, allowing for a wide range of sensitivity and responses to stressors such as metals, nutrients and sediments. Finally, they are ubiquitous and relatively easy to collect and identify which makes them attractive to agencies and organizations searching for a practical means of assessing water quality in streams.
Nutrient enrichment from livestock waste Sediment depostion from construction activity

Collecting Benthic Macroinvertebrates
Streams in West Virginia are primarily high gradient with coarse substrate materials such as boulder, cobble and gravel. These physical conditions are responsible for the typical riffle/run habitats commonly found in most streams across the state. The Watershed Assessment Branch samples these riffle/runs for benthos by using a hand-held net, often referred to as a kick-net. Generally speaking, the net is secured to the stream bottom in a riffle/run while the cobbles and gravels upstream are cleaned and disturbed ("kicked") causing the benthos to float downstream into the net. This procedure is used statewide by the WAB in all wadeable streams with riffle/run habitats and is by far the most common type of benthic sample collected by the agency. All benthos samples are taken to the laboratory where the organisms are sorted, identified and enumerated in preparation for data analysis and interpretation.

Entities wishing to collect benthic macroinvertebrates from West Virginia streams for basic environmental research studies or permitting projects will need to obtain a scientific collection permit from the West Virginia DNR, which requires that the investigator(s) follow the methods established and used by the WAB.
The WAB Standard Operating Procedures may be found following this link.
Scientific Collection Permit - Division of Natural Resources (DNR) Page
Collecting benthos Processing benthos Sorting benthos

Using Benthic Macroinvertebrates to Assess Biological Conditions
In order to extract and understand the information presented by a benthic macroinvertebrate sample, the WAB uses a tool commonly called an Index of Biotic Integrity (IBI). An IBI is a summary score that is comprised of several biological indicators called metrics. A metric is a characteristic of the biological community that changes in some predictable way with increases in human disturbance. For example, species richness (sometimes called diversity) generally exhibits a marked decrease as human disturbance increases. Therefore, it is one of the most commonly utilized component metrics of IBI's.

In 2000, the WAB collaborated with the United States Environmental Protection Agency (EPA) Region III to develop its own IBI, a multi-metric index for biological assessments called the West Virginia Stream Condition Index (WVSCI). The WVSCI summarizes family level identifications of benthic macroinvertebrate assemblages to assess the biological condition of wadeable streams with riffle/run habitats. This index includes six biological metrics that represent elements of the structure and composition of benthic macroinvertebrate communities. The WVSCI metrics were selected to maximize discrimination between streams with known stressors and reference streams. Reference streams have little or no human disturbances, and therefore exhibit good water quality and habitat conditions. Many of the streams that the WAB recognizes as reference sites are located on public lands, such as Monongahela National Forest and the West Virginia State Park system. The WAB has used the WVSCI since 2002 to assess the biological condition of wadeable streams in the state.

WVSCI Document
WVSCI Addendum

Although the family-level WVSCI was designed using sound ecological and statistical principles and has typically met the needs of the WAB, the availability of genus-level benthic macroinvertebrate data led to the development of a more accurate IBI for measuring the biological condition in wadeable streams. This IBI, known as GLIMPSS (Genus Level Index of Most Probable Stream Status), was completed in a final draft document in April 2011 and peer reviewed by a technical committee. GLIMPSS, similar to the WVSCI and other indices of biotic integrity, summarizes scores of various metrics into a single index value. The metrics were selected to maximize discrimination between streams with known stressors and reference streams. The GLIMPSS is considered an improvement over the WVSCI because it incorporates more detailed taxonomic information of benthic macroinvertebrates (genus versus family level identification), and unlike the WVSCI, is stratified and calibrated by ecoregion and by season. During its development, the GLIMPSS clearly demonstrated an improvement in detecting stressor specific impacts over the family-level WVSCI.

However, the GLIMPSS is not yet ready for use in determining attainment of a stream's Aquatic Life Use for regulatory purposes. During West Virginia's 2012 legislative session, Senate Bill 562 was passed. This legislation requires the DEP to develop new assessment methodology that will be subject to legislative approval. The process to develop and evaluate options for assessing stream health more "holistically" is ongoing, and specifically considers the use of fish community information, along with benthic macroinvertebrate index scores, as part of the assessment methodology.

WVSCI Scoring Process
The WVSCI summarizes family level identifications of benthic macroinvertebrate samples to bioassess the condition of wadeable streams. This index includes six biological metrics that represent elements of the structure and composition of benthic macroinvertebrate communities:

1. Total Taxa - Measures the total number of macroinvertebrate taxa (richness and different kinds) collected in the sample. In general, this metric exhibits a negative response to pollutants and stressors; therefore, will generally decrease as stream conditions degrade. The WVSCI uses a Total Taxa number of 22 as a best standard value. Therefore, a benthos sample with 22 or more different types of organisms would be considered very good using the WAB protocol.

2. EPT Index - Measures the total number of taxa (richness and different kinds) within the generally pollution sensitive groups: Ephemeroptera (mayflies), Plecoptera (stoneflies) and Trichoptera (caddisflies). In general, this metric exhibits a negative response to pollutants and stressors and will generally decrease as stream conditions degrade. This index is widely used because it is very sensitive to changes in water quality. The WVSCI uses an EPT Taxa number of 13 as a best standard value. Therefore, a benthos sample with 13 or more different kinds of EPT organisms would be considered very good using the WAB protocol.

3. Percent Contribution of 2 Dominant Taxa - Measures the relative abundance of the 2 numerically dominant taxa to the total number of organisms in the sample. In healthy streams, benthos are generally diverse, with the individuals being relatively evenly distributed numerically among the different taxa. As stream water quality decreases, more individuals are concentrated in fewer, more tolerant taxa, and this metric increases. The WVSCI uses 37.3% as a best standard value for this metric. Therefore, a benthos sample with 37.3% or less for the 2 Dominant Taxa metric would be considered very good using the WAB protocol.

4. Percent EPT - Measures the relative abundance of mayfly, stonefly, and caddisfly individuals to the total number of organisms in the sample. In general, this metric exhibits a negative response to pollutants and stressors and will generally decrease as stream conditions degrade. The WVSCI uses 89.3% as a best standard value for this metric. Therefore, a benthos sample equal to or greater than 89.3% would be considered very good using the WAB protocol.

5. Percent Chironomidae - Measures the relative abundance of chironomid (midges) individuals to the total number of individuals in the sample. Chironomids as a group are considered to be relatively tolerant to many pollutants and stressors. Therefore, this metric generally increases in value as stream conditions degrade. The WVSCI uses 1.7% as a best standard value for this metric.

6. HBI (Hilsenhoff's Biotic Index - modified) - Although this metric was originally developed to summarize tolerances of the benthic community to organic pollution, it has been successfully adapted for use in streams with a variety of pollutants and stressors. Tolerance values are assigned to each taxon on a scale of 0 to 10, with 0 identifying the organisms that are least tolerant (most sensitive), and 10 identifying the most tolerant (least sensitive) organisms. The HBI metric score can be thought of as an average pollution tolerance value for a sample, weighted by the abundance of organisms. As water quality of stream decreases, the HBI increases. This is especially true where organic enrichment is present. Since many of the organic pollution tolerant organisms are also tolerant to other stressors, the HBI is often used as a general indicator of stress. The WVSCI uses 2.61 as a best standard value for the HBI metric.

WVSCI- All six component metric values are converted to a standard 0 (worst) to 100 (best) point scale. The six standardized metric scores are then averaged for each benthic sample to calculate a final index score that ranges from 0.0 to 100.0. A benthic macroinvertebrate sample with a WVSCI score greater than or equal to 68.0 is considered to be good biologically. The threshold of 68.0 was derived by calculating the 5th percentile of WVSCI scores from a series of reference sites. If the score is less than 68.0, the biological condition of the stream is considered impaired and it may be placed on the West Virginia 303(d) list of impaired streams. The 303(d) list is used for Total Maximum Daily Load selection and development in the state. The examples below provide a simplification of the WVSCI analysis procedure for a reference stream that scores well above the impairment theshold, and an impaired stream that was placed on the 303(d) list.

Examples of WVSCI Calculation Procedure
Family Count     Family Count      
Cambaridae 1     Turbellaria  1      
Ameletidae 4 Analysis Procedure   Heptageniidae  2      
Heptageniidae 48 Metric Score   Ephemerellidae 5   Analysis Procedure
Leptophlebiidae 10 Taxa Richness 100   Caenidae 2   Metric Score
Ephemerellidae 3 EPT Richness 100   Perlidae 1   Taxa Richness 63.6
Nemouridae 20 %EPT 97.8   Perlodidae 11   EPT Richness 53.9
Leuctridae 21 %Chironomidae 100   Dryopidae 1   %EPT 15.9
Pteronarcyidae 13 %2 Dominant 100   Elmidae 31   %Chironomidae 34.8
Peltoperlidae 1 HBI 94.9   Psephenidae 6   %2 Dominant 32.0
Perlidae 4 Average 98.8   Rhyacophilidae 1   HBI 64.3
Perlodidae 4     Hydropsychidae 9   Average 44.1
Chloroperlidae 12     Tipulidae 3      
Elmidae 7     Simuliidae 2   WVSCI 44.1
Psephenidae 5     Chironomidae 144   Impaired
Rhyacophilidae 4              
Philopotamidae 2 WVSCI 98.8         303(d) list
Polycentropodidae 3 Not Impaired            
Hydropsychidae 11           TMDL
Lepidostomatidae 3              
Limnephilidae 4            
Uenoidae 12 Crane Creek (BST-60-F)
Panther State Forest
        Little Hurricane Creek (WVK-24)  
Tipulidae 9              
Ceratopogonidae 1              
Chironomidae 3              

Community Assessments
Fish community assessments are an important component of many water quality management programs. These assessments can be useful for making decisions in regard to biological integrity and overall stream health. Fish community surveys may prove to be a useful tool to assess Aquatic Life Use attainment in West Virginia. Currently, 2014, the WAB does not have an Index of Biotic Integrity (IBI) developed for use with the fish community data.

Consumption Advisories
Human activities have allowed the discharge of chemicals, trace metals and other contaminants into the waters of the state. Fish and other aquatic organisms may bioaccumulate these contaminants in their tissues to levels much higher than those in the water and sediments. The WAB collects fish of various species from West Virginia's waterbodies in order to determine if contaminants are present in their tissues, and to what level. This information is used by the West Virginia Department of Health and Human Resources, through an interagency agreement with West Virginia's DEP and Division of Natural Resources, to develop consumption advisories for fish caught in the state. These fish consumption advisories are reviewed annually and are geared towards helping West Virginia anglers make educated choices about eating the fish they catch.
Fish Consumption Advisory

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