Testing Bioassessment Metrics: Macroinvertebrate, Sculpin, and Salmonid Responses to Stream Habitat, Sediment, and Metals

The purpose of this article is to report on the testing of responses of multimetric macroinvertebrate and habitat indices to common disturbances to streams: stream habitat alteration, excessive sediment, and elevated metals concentrations. Seven macroinvertebrate community metrics were combined into a macroinvertebrate biotic index (MBI), and 11 channel morphology, riparian, and substrate features were combined into a habitat index. Indices were evaluated by comparing the habitat results to fish population surveys and comparing the macroinvertebrate results to habitat ratings, percent fine sediments measured by Wolman pebble counts, and copper concentrations. Macroinvertebrate scores decreased with increasing percentages of fine sediments measured either across the bankfull or instream channel widths. Macroinvertebrate scores decreased with increasing copper. One metric, richness of Ephemeroptera, Plecoptera, and Trichoptera (EPT) taxa, was more responsive to both copper and sediment than was the multimetric MBI. Habitat scores corresponded well with the age class structure of salmonids, but not with that of benthic sculpins. Both salmonid and sculpin age classes declined with increasing percentages of fine sediments. The decline was graded with the sculpin age classes, whether fine sediments were measured across the instream or bankfull channel, whereas salmonids consistently responded only to the instream fine sediments.     

The influence of suburban land use on habitat and biotic integrity of coastal Rhode Island streams

Watershed land use in suburban areas can affect stream biota through degradation of instream habitat, water quality, and riparian vegetation. By monitoring stream biotic communities in various geographic regions, we can better understand and conserve our watershed ecosystems. The objective of this study was to examine the relationship between watershed land use and the integrity of benthic invertebrate communities in eight streams that were assessed over a 3-year period (2001-2003). Sites were selected from coastal Rhode Island watersheds along a residential land-use gradient (4-59%). Using the rapid bioassessment protocol, we collected biological, physicochemical, habitat, and nutrient data from wadeable stream reaches and compared metrics of structure and integrity. Principal component analyses showed significant negative correlation of indicators for stream physicochemical, habitat, and instream biodiversity with increasing residential land use (RLU) in the watershed. The physicochemical variables that were most responsive to percent RLU were conductivity, instream habitat, nitrate, and dissolved inorganic nitrogen (DIN). The positive correlation of DIN with percent RLU indicated an anthropogenic source of pollution affecting the streams. The biotic composition of the streams shifted from sensitive to insensitive taxa as percent RLU increased; the most responsive biological variables were percent Ephemeroptera, percent Scrapers, percent Insects, and the Hilsenhoff biotic index. These data show the importance of land management and conservation at the watershed scale to sustaining the biotic integrity of coastal stream ecosystems.     

A macroinvertebrate assessment of Ozark streams located in lead–zinc mining areas of the Viburnum Trend in southeastern Missouri, USA

The Viburnum Trend lead–zinc mining subdistrict is located in the southeast Missouri portion of the Ozark Plateau. In 2003 and 2004, we assessed the ecological effects of mining in several watersheds in the region. We included macroinvertebrate surveys, habitat assessments, and analysis of metals in sediment, pore water, and aquatic biota. Macroinvertebrates were sampled at 21 sites to determine aquatic life impairment status (full, partial, or nonsupport) and relative biotic condition scores. Macroinvertebrate biotic condition scores were significantly correlated with cadmium, nickel, lead, zinc, and specific conductance in 2003 (r?=??0.61 to ?0.68) and with cadmium, lead, and pore water toxic units in 2004 (r?=??0.55 to ?0.57). Reference sites were fully supporting of aquatic life and had the lowest metals concentrations and among the highest biotic condition scores in both years. Sites directly downstream from mining and related activities were partially supporting, with biotic condition scores 10% to 58% lower than reference sites. Sites located greater distances downstream from mining activities had intermediate scores and concentrations of metals. Results indicate that elevated concentrations of metals originating from mining activities were the underlying cause of aquatic life impairment in several of the streams studied. There was general concurrence among the adversely affected sites in how the various indicators responded to mining activities during the overall study.     

Assessing the ecological condition of streams in a southeastern Brazilian basin using a probabilistic monitoring design

Prompt assessment and management actions are required if we are to reduce the current rapid loss of habitat and biodiversity worldwide. Statistically valid quantification of the biota and habitat condition in water bodies are prerequisites for rigorous assessment of aquatic biodiversity and habitat. We assessed the ecological condition of streams in a southeastern Brazilian basin. We quantified the percentage of stream length in good, fair, and poor ecological condition according to benthic macroinvertebrate assemblage. We assessed the risk of finding degraded ecological condition associated with degraded aquatic riparian physical habitat condition, watershed condition, and water quality. We describe field sampling and implementation issues encountered in our survey and discuss design options to remedy them. Survey sample sites were selected using a spatially balanced, stratified random design, which enabled us to put confidence bounds on the ecological condition estimates derived from the stream survey. The benthic condition index indicated that 62 % of stream length in the basin was in poor ecological condition, and 13 % of stream length was in fair condition. The risk of finding degraded biological condition when the riparian vegetation and forests in upstream catchments were degraded was 2.5 and 4 times higher, compared to streams rated as good for the same stressors. We demonstrated that the GRTS statistical sampling method can be used routinely in Brazilian rain forests and other South American regions with similar conditions. This survey establishes an initial baseline for monitoring the condition and trends of streams in the region.     

长江流域水生态评价指标初步研究

通过调研20世纪90年代以来国外主流水生态评价项目中采用的生境指标,统计了14个类型生境指标的使用频率,并针对不同的河流类型,筛选出物理形态特征、河岸带状况、生境组成、生境复杂性、人类干扰和水质状况等6个类型的11项评价指标,推荐作为长江流域河流生境评价指标.随后,利用层次分析法对国外水生生物评价指标进行了统计分析,并...     

Influence of Riffle and Snag Habitat Specific Sampling on Stream Macroinvertebrate Assemblage Measures in Bioassessment

Stream macroinvertebrate communities vary naturally among types of habitats where they are sampled, which affects the results of environmental assessment. We analyzed macroinvertebrates collected from riffle and snag habitats to evaluate influences of habitat-specific sampling on taxon occurrence, assemblage measures, and biotic indices. We found considerably more macroinvertebrate taxa unique to snags (143 taxa) than to riffles (75 taxa), and the numbers of taxa found in both riffles and snags (149 taxa) were similar to that found in snags. About 64% of the 47 macroinvertebrate measures we tested differed significantly between riffles and snags. Eighty percent intercepts of regressions between biotic indices and urban or agricultural land uses differed significantly between riffles and snags. The Hilsenhoff biotic index calculated from snag samples explained 69% of the variance of riffle samples and classified 66% of the sites into the same stream health group as the riffle samples. However, four multimetric indices for snag samples explained less than 50% of the variance of riffle samples and classified less than 50% of the sites into the same health group as the riffle samples. We concluded that macroinvertebrate indices developed for riffle/run habitat should not be used for snag samples to assess stream impairment. We recommend developing an index of biotic integrity specifically for snags and using snags as an alternate sampling substrate for streams that naturally lack riffles.     

Calibration and validation of a regionally and seasonally stratified macroinvertebrate index for West Virginia wadeable streams

Multimetric indices (MMIs) are routinely used by federal, state, and tribal entities to assess the quality of aquatic resources. Because of their diversity, abundance, ubiquity, and sensitivity to environmental stress, benthic macroinvertebrates are well suited for MMIs. West Virginia has used a statewide family-level stream condition index (WVSCI) since 2002. We describe the development, validation, and application of a geographically- and seasonally partitioned genus-level index of most probable stream status (GLIMPSS) for West Virginia wadeable streams. Natural classification strata were evaluated with reference site communities using mean similarity analysis and non-metric multidimensional scaling ordination. Forty-one metrics spanning six ecological categories (richness, composition, tolerance, dominance, trophic groups, and habits) were evaluated for sensitivity, responsiveness, redundancy, range and variability across seasonal (spring and summer) and regional (mountains and plateau) strata. Through a step-wise metric selection process, 8–10 metrics were aggregated to comprise four stratum-specific GLIMPSS models (mountain/plateau and spring/summer). A comparison of GLIMPSS with WVSCI exhibited marked improvements where GLIMPSS detecting greater stream impacts. A variation of the GLIMPSS, which differs only in the family-level taxonomic identification of Chironomidae (GLIMPSS (CF)), was comparable to the full GLIMPSS. These MMIs are robust yet practical tools for evaluating impacts to water quality, instream and riparian habitat, and aquatic wildlife in wadeable riffle-run streams based on sensitivity, responsiveness, precision, and independent validation. These models may be used effectively to detect degradation of the naturally occurring benthic community, assess causes of biological degradation, and plan and evaluate remediation of damaged stream ecosystems.     

Development of a Provisional Physical Habitat Index for Maryland Freshwater Streams

Provisional physical habitat indices were developed and validatedfor Maryland Coastal and Non-Coastal Plain streams using variables (commonly called metrics) that best discriminated reference and degraded conditions based on biological, chemicaland land use data from the 1994–97 Maryland Biological Stream Survey (MBSS). These habitat indices contained variables that described structural, hydrological, vegetative and aesthetic components of stream habitat. Variables with the best discriminatory power for Coastal Plain streams were: instream habitat, velocity/depth diversity, pool/glide/eddy quality, embeddedness, maximum depth and aesthetic rating. Physical habitat variables with the best discriminatory power for Non-Coastal Plain sites were: instream habitat, velocity/depth diversity, riffle/run quality, embeddedness, number of rootwads and aesthetic rating. The overall classification efficiency forindex validation was 76% for both indices pooled over both strata. Scaled physical habitat index values (0–100) for bothstrata identified nearly twice as many good sites (31%) as very poor sites (16%). More than half the Maryland sites werein the poor to fair range (53%).     

Maryland Biological Stream Survey: A State Agency Program to Assess the Impact of Anthropogenic Stresses on Stream Habitat Quality and Biota

The Maryland Department of Natural Resources is conducting the Maryland Biological Stream Survey, a probability-based sampling program, stratified by river basin and stream order, to assess water quality, physical habitat, and biological conditions in first through third order, non-tidal streams. These streams comprise about 90% of all lotic water miles in the state. About 300 sites (75 m segments) are being sampled during spring and summer each year. All basins in the state will be sampled over a three-year period, 1995-97. MBSS developments in 1995-96 included (1) an electrofishing capture efficiency correction method to improve the accuracy of fish population estimates, (2) two indices of biotic integrity (IBI) for fish assemblages to identify degraded streams, and (3) land use information for catchments upstream of sampled sites to investigate associations between stream condition and anthropogenic stresses. Based on fish IBI scores at 270 stream sites in six basins sampled in 1995, 11% of non-tidal stream miles in Maryland were classified as very poor, 15% as poor, 24% as fair, and 27% as good. IBIs have not yet been developed for stream sites with catchment areas less than 120 hectares (23% of non-tidal stream miles). IBI scores declined with stream acid neutralizing capacity (ANC) and pH, an association that was also evident for fish species richness, biomass, and density. Low IBI scores were associated with several measures of degraded stream habitat, but not with local riparian buffer width. There was a significant negative association between IBI scores and urban land use upstream of sampled sites in the only extensively urbanized basin assessed in 1995. Future plans for the MBSS include (1) identifying all benthic macroinvertebrate samples to genus, (2) developing benthic macroinvertebrate, herpetofaunal, and physical habitat indicators, and (3) enhancing the analysis of stream condition-stressor associations by refining landscape metrics and using multi-variate techniques.     

Hydrologic disturbance reduces biological integrity in urban streams

The impact of urbanization on stream ecosystems is linked by land cover changes to the alteration of the natural hydrology and subsequent physical disruption of stream biota and habitat. Seasonal floods are part of the natural disturbance regime of many streams, but urbanization increases their frequency and magnitude. This study evaluated the impact of hydrologic disturbance on fish and aquatic macroinvertebrates in 81 (56 urban/25 reference) Ohio streams. Hydrologic variables included annual and monthly 24-h rainfall maxima and computed annual peak discharge, with computation supported by GIS-based drainage area delineation and land cover characterization. Ohio biological criteria for fish and macroinvertebrates measured during the late spring and summer were negatively impacted by annual peak discharge in urban streams as compared to reference streams. Results support the application of stormwater best management practices as part of stream restoration efforts to mitigate urbanization impacts to fish and macroinvertebrates.     

Spatial heterogeneity of stream environmental conditions and macroinvertebrates community in an agriculture dominated watershed and management implications for a large river (the Liao River,China) basin

Understanding the effects of watershed land uses (e.g., agriculture, urban industry) on stream ecological conditions is important for the management of large river basins. A total of 41 and 56 stream sites (from first to fourth order) that were under a gradient of watershed land uses were monitored in 2009 and 2010, respectively, in the Liao River Basin, Northeast China. The monitoring results showed that a total of 192 taxa belonging to four phyla, seven classes, 21 orders and 91 families were identified. The composition of macroinvertebrate community in the Liao River Basin was dominated by aquatic insect taxa (Ephemeroptera and Diptera), Oligochaeta and Molluscs. The functional feeding group GC (Gatherer/Collector) was dominant in the whole basin. Statistical results showed that sites with less watershed impacts (lower order sites) were characterized by higher current velocity and habitat score, more sensitive taxa (e.g., Ephemeroptera), and the substrate was dominated by high percentage of cobble and pebble. The sites with more impacts from agriculture and urban industry (higher order sites) were characterized by higher biochemical (BOD₅) and chemical oxygen demand (COD), more tolerant taxa (e.g., Chironominae), and the substrate was dominated by silt and sand. Agriculture and urban-industry activities have reduced habitat condition, increased organic pollutants, reduced macroinvertebrate abundance, diversity, and sensitive taxa in streams of the lower Liao River Basin. Restoration of degraded habitat condition and control of watershed organic pollutants could be potential management priorities for the Basin.     

Evaluation of an alternate method for sampling benthic macroinvertebrates in low-gradient streams sampled as part of the National Rivers and Streams Assessment

Benthic macroinvertebrates are sampled in streams and rivers as one of the assessment elements of the US Environmental Protection Agency’s National Rivers and Streams Assessment. In a 2006 report, the recommendation was made that different yet comparable methods be evaluated for different types of streams (e.g., low gradient vs. high gradient). Consequently, a research element was added to the 2008–2009 National Rivers and Streams Assessment to conduct a side-by-side comparison of the standard macroinvertebrate sampling method with an alternate method specifically designed for low-gradient wadeable streams and rivers that focused more on stream edge habitat. Samples were collected using each method at 525 sites in five of nine aggregate ecoregions located in the conterminous USA. Methods were compared using the benthic macroinvertebrate multimetric index developed for the 2006 Wadeable Streams Assessment. Statistical analysis did not reveal any trends that would suggest the overall assessment of low-gradient streams on a regional or national scale would change if the alternate method was used rather than the standard sampling method, regardless of the gradient cutoff used to define low-gradient streams. Based on these results, the National Rivers and Streams Survey should continue to use the standard field method for sampling all streams.     

Bioassessment tools in novel habitats: an evaluation of indices and sampling methods in low-gradient streams in California

Biomonitoring programs are often required to assess streams for which assessment tools have not been developed. For example, low-gradient streams (slope ≤1%) comprise 20–30% of stream miles in California and are of particular interest to watershed managers, yet most sampling methods and bioassessment indices in the state were developed in high-gradient systems. This study evaluated the performance of three sampling methods [targeted riffle composite (TRC), reach-wide benthos (RWB), and the margin–center–margin modification of RWB (MCM)] and two indices [the Southern California Index of Biotic Integrity (SCIBI) and the ratio of observed to expected taxa (O/E)] in low-gradient streams in California for application in this habitat type. Performance was evaluated in terms of efficacy (i.e., ability to collect enough individuals for index calculation), comparability (i.e., similarity of assemblages and index scores), sensitivity (i.e., responsiveness to disturbance), and precision (i.e., ability to detect small differences in index scores). The sampling methods varied in the degree to which they targeted macroinvertebrate-rich microhabitats, such as riffles and vegetated margins, which may be naturally scarce in low-gradient streams. The RWB method failed to collect sufficient numbers of individuals (i.e., ≥450) to calculate the SCIBI in 28 of 45 samples and often collected fewer than 100 individuals, suggesting it is inappropriate for low-gradient streams in California; failures for the other methods were less common (TRC, 16 samples; MCM, 11 samples). Within-site precision, measured as the minimum detectable difference (MDD) was poor but similar across methods for the SCIBI (ranging from 19 to 22). However, RWB had the lowest MDD for O/E scores (0.20 versus 0.24 and 0.28 for MCM and TRC, respectively). Mantel correlations showed that assemblages were more similar within sites among methods than within methods among sites, suggesting that the sampling methods were collecting similar assemblages of organisms. Statistically significant disagreements among methods were not detected, although O/E scores were higher for RWB samples than TRC. Index scores suggested impairment at all sites in the study. Although index scores did not respond strongly to several measurements of disturbance in the watershed, percent agriculture showed a significant, negative relationship with O/E scores.     

Montane meadows in the Sierra Nevada: comparing terrestrial and aquatic assessment methods

We surveyed montane meadows in the northern Sierra Nevada and southern Cascades for two field seasons to compare commonly used aquatic and terrestrial-based assessments of meadow condition. We surveyed (1) fish, (2) reptiles, (3) amphibians, (4) aquatic macroinvertebrates, (5) stream geomorphology, (6) physical habitat, and (7) terrestrial vegetation in 79 meadows between the elevations of 1,000 and 3,000?m. From the results of those surveys, we calculated five multi-metric indices based on methods commonly used by researchers and land management agencies. The five indices consisted of (1) fish only, (2) native fish and amphibians, (3) macroinvertebrates, (4) physical habitat, and (5) vegetation. We compared the results of the five indices and found that there were significant differences in the outcomes of the five indices. We found positive correlations between the vegetation index and the physical habitat index, the invertebrate index and the physical habitat index, and the two fish-based indices, but there were significant differences between indices in both range and means. We concluded that the five indices provided very different interpretations of the condition in a given meadow. While our assessment of meadow condition changed based on which index was used, each provided an assessment of different components important to the overall condition of a meadow system. Utilizing a multimetric approach that accounts for both terrestrial and aquatic habitats provides the best means to accurately assess meadow condition, particularly given the disproportionate importance of these systems in the Sierra Nevada landscape.     

The effects of landscape-level disturbance on the composition of Minnesota caddisfly (Insecta: Trichoptera) trophic functional groups: evidence for ecosystem homogenization

Over 300,000 caddisfly specimens representing 249 species were collected from nearly 250 sites throughout Minnesota during 2000 and 2001 to determine the effects of human disturbance on the composition of caddisfly trophic functional groups at the landscape level. Canonical correspondence analysis determined that stream width was the most important variable influencing functional group composition in regions of the state with relatively low disturbance, and that differences in the caddisfly fauna between sizes of streams generally followed trends predicted by the river continuum concept. In regions of the state with moderate disturbance, both stream width and the percentage of disturbed habitat upstream of a site were important variables influencing functional group composition. In highly disturbed regions, no variables corresponded to changes in the composition of caddisfly functional groups. Instead, ecosystems were homogeneous: fine-particle filtering collectors dominated in all sizes of streams. The observed aquatic ecosystem homogenization is attributed mostly to input of fine-particle organic and inorganic sediment from extensive agriculture.     

A Fish-Based Index of Biotic Integrity to Assess Intermittent Headwater Streams in Wisconsin, USA

I developed a fish-based index of biotic integrity (IBI) to assess environmental quality in intermittent headwater streams in Wisconsin, USA. Backpack electrofishing and habitat surveys were conducted four times on 102 small (watershed area 1.7–41.5 km²), cool or warmwater (maximum daily mean water temperature ≥22 C), headwater streams in spring and late summer/fall 2000 and 2001. Despite seasonal and annual changes in stream flow and habitat volume, there were few significant temporal trends in fish attributes. Analysis of 36 least-impacted streams indicated that fish were too scarce to calculate an IBI at stations with watershed areas less than 4 km² or at stations with watershed areas from 4–10 km² if stream gradient exceeded 10 m/km (1% slope). For streams with sufficient fish, potential fish attributes (metrics) were not related to watershed size or gradient. Seven metrics distinguished among streams with low, agricultural, and urban human impacts: numbers of native, minnow (Cyprinidae), headwater-specialist, and intolerant (to environmental degradation) species; catches of all fish excluding species tolerant of environmental degradation and of brook stickleback (Culaea inconstans) per 100 m stream length; and percentage of total individuals with deformities, eroded fins, lesions, or tumors. These metrics were used in the final IBI, which ranged from 0 (worst) to 100 (best). The IBI accurately assessed the environmental quality of 16 randomly chosen streams not used in index development. Temporal variation in IBI scores in the absence of changes in environmental quality was not related to season, year, or type of human impact and was similar in magnitude to variation reported for other IBI's.     

Integrative Evaluation of the Ecological Quality of Metalliferous Streams (Chalkidiki, Macedonia, Hellas)

The present study reports on a year-long (November 1997-November 1998) study of water quality in streams of the Olympias and Skouries areas of Chalkidiki (Hellas), subject to past and present mining activities. Benthic macroinvertebrates were collected from 18 sampling sites at five sampling intervals, while several physicochemical parameters of the water (D.O., pH, temperature, nutrients etc.) were also measured. Five European biotic indices and scores and two multivariate statistical methods (CANOCO, FUZZY) were applied to the macroinvertebrate data. The concentration of various trace metals in the water, sediment, fish and crustaceans were also measured. Most sampling stations exhibited a highly diverse macroinvertebrate fauna during the whole year. However, a slight amelioration of water quality was observed during the high-flow period (winter and early spring), regarding diversity of several invertebrate groups, especially pollution-sensitive families, which decreased. The European biotic indices and scores proved to be relatively ineffective in describing water quality and it is suggested that an Hellenic biotic indice or score should be created. The bioaccumulation studies indicated variable effects of trace metals and organic load on macroinvertebrates according to the degree of load.     

Relationships among nekton assemblage structure and abiotic conditions in three Texas tidal streams

The objective of this study was to assess the effects of spatially and temporally varying abiotic conditions on the nekton residing in tidally influenced streams and determine the comparability of the assemblage response across the various gears needed to fully characterize the biota. Three tidal streams were sampled seasonally for 2 years for chemical (physiochemical profiles), physical (in-stream and riparian habitat classification), and biological (nekton sampled with bag seines, trawls, and gill nets) components of ecosystem integrity. Multiple sampling stations on each stream encompassed the transitional character of the tidally influenced ecosystem, from the freshwater of the river to the saltwater of the bay. Instead of characterizing the biota with traditional fish-based indexes of biotic integrity, analysis methodologies relying heavily on multivariate ordination techniques were used. This study showed that the temporal and spatial relationships among nekton assemblages and abiotic environmental conditions were quite gear dependent. The greatest degree of difference in indicators of ecosystem health all involved upstream–downstream gradients that appear to be driven by salinity structure. Based on the results of this study, dissolved oxygen concentration does not appear to be a major structuring factor in the physical, chemical, or biological component of ecosystem integrity.     

Comparison of Streambed Sediment and Aquatic Biota as Media for Characterizing Trace Elements and Organochlorine Compounds in the Willamette Basin, Oregon

During 1992–93, 27 organochlorine compounds (pesticides plus total PCB) and 17 trace elements were analyzed in bed sediment and aquatic biota from 20 stream sites in the Willamette Basin as part of the U.S. Geological Survey's National Water-Quality Assessment Program. Data from each medium were compared to evaluate their relative effectiveness for assessing occurrence (broadly defined as documentation of important concentrations) of these constituents. Except for Cd, Hg, Se, and Ag, trace element concentrations generally were higher in bed sediment than in biota. Conversely, although frequencies of detection for organochlorine compounds in biota were only slightly greater than in bed sediment, actual concentrations in biota (normalized to lipid) were as much as 19 times those in sediment (normalized to organic carbon). Sculpin (Cottus spp.) and Asiatic clams (Corbicula fluminea), found at 14 and 7 sites, respectively, were the most widespread taxa collected during the study. Concentrations of trace elements, particularly As and Cu, were typically greater in Asiatic clams than in sculpin. In contrast, almost half of the organochlorine compounds analyzed were found in sculpin, but only DDT and its degradation products were detected in Asiatic clams; this may be related to the lipid content of sculpin, which was about three times higher than for clams. Thus, the medium of choice for assessing occurrence depends largely on the constituent(s) of interest.     

Biomonitoring of Lead, Zinc, and Cadmium in Streams Draining Lead-Mining and Non-Mining Areas, Southeast Missouri, USA

We evaluated exposure of aquatic biota to lead (Pb), zinc (Zn), and cadmium (Cd) in streams draining a Pb-mining district in southeast Missouri. Samples of plant biomass (detritus, periphyton, and filamentous algae), invertebrates (snails, crayfish, and riffle benthos), and two taxa of fish were collected from seven sites closest to mining areas (mining sites), four sites further downstream from mining (downstream sites), and eight reference sites in fall 2001. Samples of plant biomass from mining sites had highest metal concentrations, with means 10- to 60-times greater than those for reference sites. Mean metal concentrations in over 90% of samples of plant biomass from mining sites were significantly greater than those from reference sites. Mean concentrations of Pb, Zn, and Cd in most invertebrate samples from mining sites, and mean Pb concentrations in most fish samples from mining sites, were also significantly greater than those from reference sites. Concentrations of all three metals were lower in samples from downstream sites, but several samples of plant biomass from downstream sites had metal concentrations significantly greater than those from reference sites. Analysis of supplemental samples collected in the fall of 2002, a year of above-average stream discharge, had lower Pb concentrations and higher Cd concentrations than samples collected in 2001, near the end of a multi-year drought. Concentrations of Pb measured in fish and invertebrates collected from mining sites during 2001 and 2002 were similar to those measured at nearby sites in the 1970s, during the early years of mining in the Viburnum Trend. Results of this study demonstrate that long-term Pb mining activity in southeast Missouri has resulted in significantly elevated concentrations of Pb, Cd, and Zn in biota of receiving streams, compared to biota of similar streams without direct influence of mining. Our results also demonstrate that metal exposure in the study area differed significantly among sample types, habitats, and years, and that these factors should be carefully considered in the design of biomonitoring studies.