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Ecological Exposure Research Division Publications: 2000

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This page lists publication titles, citations and abstracts produced by NERL's Ecological Exposure Research Division for the year 2000, organized by Publication Type. Your search has returned 41 Matching Entries.

See also Ecological Exposure Research Division citations with abstracts: 1999,  2000,  2001,  2002,  2003,  2004,  2005,  2006,  2007,  2008,  2009

Technical Information Manager: Linda Ransick - (513) 569-7395 or ransick.linda@epa.gov

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Presented/Published
JOURNAL Microsatellite Characterization in Central Stoneroller Campostoma Anomalum (Pisces: Cyprinidae) 12/01/2000
Dimsoski, P., G P. Toth, AND M J. Bagley. Microsatellite Characterization in Central Stoneroller Campostoma Anomalum (Pisces: Cyprinidae). MOLECULAR ECOLOGY 9(12):2187-2189, (2000).
Abstract: The central stoneroller (Campostoma anomalum) is a small cyprinid fish that is native to streams and rivers of central and eastern North America. It can be found in a range of anthropo- genically modified habitats, ranging from nearly pristine to highly polluted waters (Zimmerman et al. 1980), and has intermediate sensitivity to habitat degradation relative to other fishes in the region (Zimmerman et al. 1980, Gillespie and Guttman 1989). The species is the focus of intensive study by the United States Environmental Protection Agency due to its biological and distributional characteristics. An important aspect of this research is to understand the fine-scale genetic structure of the species across its native range and to determine how this "genetic landscape" relates to underlying enviornmental processes. To date, genetic analyses have focused on multi-locus fingerprints generated by the RAPD method to delineate levels of similarity among and within populations (Silbiger et al. 1998). Because allelic counts are highly sensitive to recent changes in population size, highly polymorphic microsatellite DNA markers should provide genetic information that is highly complementary to the RAPD data and may reveal finer levels of populations structuring. Here, we report a suite of highly polymorphic microsatellite markers developed for the central stoneroller.

JOURNAL Method-Specific Precision and Bias Relationships Developed from Data Submitted During USEPA Wastewater Laboratory Performance Evaluation Studies 11/01/2000
Britton, P W. Method-Specific Precision and Bias Relationships Developed from Data Submitted During USEPA Wastewater Laboratory Performance Evaluation Studies. JOURNAL OF TESTING AND EVALUATION (ASTM) 28(6):526-546, (2000).
Abstract: This paper documents the process used by the United States Environmental Protection Agency (USEPA) to estimate the mean and standard deviation of data reported by in-control wastewater laboratories during Water Pollution (WP) studies. This process is then applied to the data reported for each specific sample, analyte, and method in ten WP studies and linear relationships between the mean response and true analyte concentration, and between the interlaboratory standard deviation of responses and true analyte concentration, are presented. Such relationships can be invaluable as background information for anyone who must conduct an interlaboratory study using the same methods to analyze similar interference-free samples, or who must evaluate the analytical quality of such data.

JOURNAL Comparison of Geographic Classification Schemes for Mid-Atlantic Stream Fish Assemblages 09/01/2000
McCormick, F H., D V. Peck, AND D P. Larsen. Comparison of Geographic Classification Schemes for Mid-Atlantic Stream Fish Assemblages. JOURNAL OF THE NORTH AMERICAN BENTHOLOGICAL SOCIETY 19(3):385-404, (2000).
Abstract: Understanding the influence of geographic factors in structuring fish assemblages is crucial to developing a comprehensive assessment of stream conditions. We compared the classification strengths (CS) of geographic groups (ecoregions and catchments), stream order, and groups based on cluster analysis of fish assemblage data from 200 wadeable streams in the Mid-Atlantic Highlands. We first calculated intersite similarity indices (Bray-Curtis on relative abundance; Dice-Sorensen on presence/absence), then compared overall mean within-group similarities (W) with among-group similarities (B). We used subsets of the data to test CS from(1) 31 reference sites defined on chemistry and habitat criteria, and (2) 21 samples from 8 sites that had been resampled within and between years to estimate the maximum similarity expected for any of the classifications. We assessed the strength of each classification by determining the degree to which W was greater than B. Sites classified by taxonomic clusters had higher CS than did sites grouped by stream order, U.S. Geological Survey 4-digit Hydrologic Unit Code (HUC) catchments, and ecoregions. Except for taxonomic clusters, the CS values were greater when all sites were used in the analysis than when only reference sites were used. The mean similarities for the revisits were 2-3 times greater than for all other classifications. We used nonmetric multidimensional scaling as an alternative approach to detecting geographic structure in the data. We found little separation of ecoregion or catchment groups except at very broad spatial scales. The relatively weak CS of any of the geographic groups suggests that the interaction of complex zoogeographic patterns and a long history of human disturbance has masked any fine-scale structure of regional fish assemblages.

JOURNAL Comparing the Strengths of Geographic and Non-Geographic Classifications of Stream Benthic Macroinvertebrates in the Mid-Atlantic Highlands, USA 08/01/2000
Waite, I. R., A. T. Herlihy, D P. Larsen, AND D J. Klemm. Comparing the Strengths of Geographic and Non-Geographic Classifications of Stream Benthic Macroinvertebrates in the Mid-Atlantic Highlands, USA. JOURNAL OF THE NORTH AMERICAN BENTHOLOGICAL SOCIETY 19(3):429-441, (2000).
Abstract: The US Environmental Protection Agency's (USEPA) Environmental Monitoring and Assessment Program (EMAP) sampled approximately 500 wadeable streams in the Mid-Atlantic Highlands region of the US during the late spring of 1993 to 1995 for a variety of physical, chemical and biological indicators of environmental condition. Eighty-eight sites that were minimally affected by human activities were chosen to determine the extent to which geographic and stream-classifications accounted for variation in the composition of riffle macroinvertebrate assemblages. Bray-Curtis similarities among sites were calculated from the relative abundance of macroinvertebrates to assess the strength of classifications based on geography (ecoregions and catchments), habitat (slope and stream order), and water chemistry (conductivity). For comparison, a taxonomic classification (two-way indicator species analysis, TWINSPAN) and a gradient analysis (correspondence analysis, CA) were performed on the macroinvertebrate data. To assess the effect of taxonomic resolution, all analyses were completed at the family level and to lowest practical taxon. The large overall variation within and among ecoregions resulted in a low average classification strength (CS) of ecoregions, although some ecoregions had high CS. Stream order had the highest CS of the habitat and water chemistry classifications. Ecoregion CS increased, however, when stream sites were first stratified by stream order (ecoregions nested within stream order). Nested ecoregion CS did not increase within first order streams, yet increased within second and third order streams. CA ordinations and TWINSPAN classification showed a clear gradient of streams along stream size (order), with a clear separation of first and third order streams based on macroinvertebrate composition. The ordinations did not, however, show a distinct clustering of sites on the basis of ecoregions. Overall, the lowest practical taxon level of identification resulted in a clearer pattern of sites in ordination space than did family-level identification, yet only a slight improvement in the different classifications (geographic, habitat, and water chemistry) based on average similarity.

JOURNAL Interregional Comparisons of Sediment Microbial Respiration in Streams 06/01/2000
Hill, B H., R. K. Hall, P. Husby, A. T. Herlihy, M. Dunne, AND M. Dunne. Interregional Comparisons of Sediment Microbial Respiration in Streams. FRESHWATER BIOLOGY 44(2):213-222, (2000).
Abstract: The rate of microbial respiration on fine-grained stream sediments was measured at 369 first to fourth-order streams in the Central Appalachians, Colorado's Southern Rockies, and California's Central Valley in 1994 and 1995. Study streams were randomly selected from the United States Environmental Protection Agency's (USEPA) River Reach File (RF3) using the sample design developed by USEPA's Environmental Monitoring and Assessment Program (EMAP). Respiration rate ranged from 0 to 0.621 g 02 g-1 AFDM h-1 in Central Appalachian streams, 0-254 g 02 g-1 AFDM h-1 in Southern Rockies streams, and 0-0.436 g 02 g-1 AFDM h-1 in Central Valley streams. Respiration was significantly lower in Southern Rocky Mountain streams and in cold water streams of the Central Appalachians. Within a defined index period, respiration was not significantly different between years, and was significantly correlated with stream temperature and chemistry (DOC, total N, total P, K, Cl, and alkalinity). The uniformity of respiration estimates among the three study regions suggests that sediment microbial respiration may be collected at any number of scales above the site-level for reliable prediction of respiration patterns at larger spatial scales.

JOURNAL Ecoregions and Benthic Diatom Assemblages in Mid-Atlantic Highlands Streams 06/01/2000
Pan, Y., R. J. Stevenson, B H. Hill, A. T. Herlihy, AND A. T. Herlihy. Ecoregions and Benthic Diatom Assemblages in Mid-Atlantic Highlands Streams. JOURNAL OF THE NORTH AMERICAN BENTHOLOGICAL SOCIETY 19(3):518-540, (2000).
Abstract: Ecoregional differences in geology and hydrology may affect physical and chemical conditions in streams and, consequently, the species composition of algal assemblages. Stresses resulting from human disturbance, however, may constrain species membership in algal assemblages and reduce regional diversity. We expected that ecoregional differences in diatom assemblages, if they were present, would be more evident in relatively undisturbed sites than in randomly selected sites. Benthic diatom and water chemistry samples were collected from streams in 7 ecoregions of the Mid-Atlantic Highlands to evaluate correspondence between ecoregional classification and diatom assemblages. Ecoregional differences were assessed using 196 randomly selected stream sites (probability sites) and 60 sites with less disturbance by humans (reference sites). Multivariate analyses showed that significant ecoregional differences in diatom assemblages were observed only in probability sites and not in reference sites. Water chemistry was significantly different among ecoregions, both for probability sites and for reference sites. Significant differences in diatom assemblages and water chemistry were, however, evident only among ecoregions grouped by topography (i.e., montane, high plateau, and low plateau/valley). Ecoregional differences between montane regions or low plateau/valley regions were subtle. Stream sites grouped by catchments were also significantly different in water chemistry but not in diatom assemblages, both for probability sites and for reference sites. Our data suggest that diatom assemblages respond to land use, especially agricultural activities, and thus may correspond to the ecoregional classification when land use differs significantly among these ecoregions (e.g., montane vs valley ecoregions). Diatom assemblages that lack a region-specific feature may be ideal as unbiased indicators of stream water quality.

JOURNAL Erpobdella Lahontana (Annelida: Hirudinea: Arhynchobdellida: Erpobdellidae), a New Species of Freshwater Leech from North America 04/11/2000
Hovingh, P., D. Klemm, AND D J. Klemm. Erpobdella Lahontana (Annelida: Hirudinea: Arhynchobdellida: Erpobdellidae), a New Species of Freshwater Leech from North America. PROCEEDINGS OF THE BIOLOGICAL SOCIETY OF WASHINGTON 113(1):155-161, (2000).
Abstract: New species of a leech, Erpobdella lahontana, is described from the Lahontan Basin in California and Nevada of the western United States. This species has four pairs of eyes, the preatrial loops of male paired ducts extend to ganglion XI, and the male and female gonopores are located in furrows of the auli, separated by five annuli.

JOURNAL Using Historical Biological Data to Evaluate Status and Trends in the Big Darby Creek (Ohio) Watershed 04/01/2000
SchubauerBerigan, M, M. Smith, J Hopkins, AND S M. Cormier. Using Historical Biological Data to Evaluate Status and Trends in the Big Darby Creek (Ohio) Watershed. ENVIRONMENTAL TOXICOLOGY AND CHEMISTRY 19(4):1097 - 1105, (2000).
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JOURNAL Quality Assurance Considerations for Use of the Fluorimager Si and Fragment Analysis Software 03/01/2000
Christ, S A., R N. Silbiger, M. Garg, S E. Franson, AND G P. Toth. Quality Assurance Considerations for Use of the Fluorimager Si and Fragment Analysis Software. ELECTROPHORESIS 21(5):874-888, (2000).
Abstract: The Fluorimager SI (FSI) from Molecular Dynamics is one of several scanning instruments available for the detection of fluorescent emissions associated with DNA samples in a variety of matrices (agarose and polyacrylamide gels, membranes and microplates). In our laboratory, we measured the electrophoretic moblity of randomly amplified polymorphic DNA (RAPD) fragments stained with ethidium bromide in agarose using the FSI to scan gels and the associated Molecular Dynamics software (ImageQuaNT and FragmeNT Analysis) for analysis. Initial scans and analyses resulted in inconsistent band detection across the same gel and across several scans of the same gel. To determine the best types of calibration for the instrument, several factors were considered and then evaluated. Tests of calibration acceptability were also evaluated. Band detection by FragmeNT Analysis was improved following optimization of matrices and parameters used in calibration and experimental scans. In addition, use of software templates for analysis and modifications in the staining procedure, which have resulted in decreased instrument associated variance, are discussed.

JOURNAL Temporal Trends in Ethoxyresorufin-O-Deethylase Acitivity of Brook Trout (Salvelinus Fontinalis) Fed 2,3,7,8-Tetrachlorodibenzo-P-Dioxin 02/01/2000
Cormier, S M., M. R. Millward, C. Mueller, B Subramanian, R D. Johnson, AND J E. Tietge. Temporal Trends in Ethoxyresorufin-O-Deethylase Acitivity of Brook Trout (Salvelinus Fontinalis) Fed 2,3,7,8-Tetrachlorodibenzo-P-Dioxin. C.H. Ward (ed.), ENVIRONMENTAL TOXICOLOGY AND CHEMISTRY. SETAC Press, Pensacola, FL, 19(2):462-471, (2000).
Abstract: Changes in ethoxyresorufin-0-deethylase (EROD) activity were monitored through an extended 6-month dietary exposure to determine the relationship between EROD activity and uptake of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in brook trout, Salvelinus fontinalis. Brook trout were fed labeled TCDD during a 4-week loading phase and an 11-week maintenance phase to achieve whole-body concentrations of 0,75, 150, 300, 600, and 1,200 pg TCDD/g fish. A spawning phase followed during which no TCDD was introduced. The TCDD had an extended half-life, with maximal levels detected in the late loading-early maintenance phases and 81 d after TCDD had been removed from the diet. Accumulation in liver increased as whole-body target concentration increased but was generally less than half of anticipated whole-body target concentrations. The EROD activity demonstrated a dose-dependent increase. Positive correlations were observed between EROD activity and TCDD body burdens for both males and females. For males, maximal induction was attained early in the maintenance phase and maintained during latter phases. For females, induction was characterized by a biphasic pattern. Maximal induction was attained during late loading-early maintenance, with an attenuated ressponse observed just before spawning. In addition, the induction response was modulated by sex, as induction was lower in females when compared with males. If sexual biases are considered, increased EROD acitvity may serve as an indicator of level of TCDD exposure and a sublethal predictor of effects of exposure.

JOURNAL Heavy Metals Structure Benthic Comunities in Colorado Mountain Streams 02/01/2000
Clements, W. H., D. M. Carlisle, J M. Lazorchak, AND P. C. Johnson. Heavy Metals Structure Benthic Comunities in Colorado Mountain Streams. ECOLOGICAL APPLICATIONS 10(2):626-638, (2000).
Abstract: The development of field sampling designs that employ multiple reference and polluted sites has been proposed as an alternative to the traditional upstream vs. downstream approach used in most biomonitoring studies. Spatially extensive monitoring programs can characterize ecological conditions within an ecoregion and provide the necessary background information to evaluate future changes in water quality. We measured physicochemical characteristics, heavy-metal concentrations, and benthic macroinvertebrate community structure at 95 sites in the Southern Rocky Mountain ecoregion in Colorado, USA. Most sites (82%) were selected using a systematic, randomized sampling design. Each site was placed into one of four metal categories (background, low, medium, and high metals), based on the cumulative criterion unit (CCU), which we defined as the ratio of the instream metal concentration to the U.S. Environmental Protection Agency criterion concentration, summed for all metals measured. A CCU of 1.0 represents a conservative estimate of the total metal concentration that, when exceeded, is likely to cause harm to aquatic organisms. Although the CCU was less than 2.0 at most (66.3%) of the sites, values exceeded 10.0 at 13 highly polluted stations. Differences among metal categories were highly significant for most measures of macroinvertebrate abundance and all measures of species richness. We observed the greatest effects on several species of heptageniid mayflies (Ephemeroptera: Heptageniidae), which were highly sensitive to heavy metals and were reduced by >75% at moderately polluted stations. The influence of taxonomic aggregation on responses to metals was also greatest for mayflies. In general, total abundance of mayflies and abundance of heptageniids were better indicators of metal pollution than abundance of dominant mayfly taxa. We used stepwise multiple-regression analyses to investigate the relationship between benthic community measures and physiochemical characteristics at the 78 randomly selected sites. Heavy-metal concentration was the most important predictor of benthic community structure at these sites. Because of the ubiquitous distribution of heavy-metal pollution in the Southern Rocky Mountain ecoregion, we conclude that potential effects of heavy metals should be considered when investigating large-scale spatial patterns of benthic macroinvertebrate communities in Colorado's mountain streams.

JOURNAL Bioequivalence Approach for Whole Effluent Toxicity Testing 01/01/2000
Shukla, R., Q. Wang, F A. Fulk, C. Deng, AND D. Denton. Bioequivalence Approach for Whole Effluent Toxicity Testing. ENVIRONMENTAL TOXICOLOGY AND CHEMISTRY 19(1):169-174, (2000).
Abstract: Increased use of whole effluent toxicity (WET) tests in the regulatory arena has brought increased concern over the statistical analysis of WET test data and the determination of toxicity. One concern is the issue of statistical power. A number of WET tests may pass the current hypothesis test approach because they lack statistical power to detect relevant toxic effects because of large within-test variability. Additionally, a number of WET tests may fail the current approach because they possess excessive statistical power, as a result of small within-test variability, and detect small differences that may not be biologically relevant. The strengths and limitations of both the traditional hypothesis test approach and the bioequivalence approach for use in the National Pollutant Discharge Elimination System program were evaluated. Data from 5,213 single-concentration, short-term chronic WET tests with Ceriodaphnia dubia provided the database for analysis. Comparison of results between the current approach and the bioequivalence approach indicates that the current approach to WET testing is generally sound but that adopting the proposed bioequivalence approach resolves concerns of statistical power. Specifically, within this data set, applying the bioequivalence approach resulted in failure for tests with relatively large test variability and a pass for tests with relatively small within-test variablity.

JOURNAL Using Regional Exposure Criteria and Upstream Reference Data to Characterize Spatial and Temporal Exposures to Chemical Contaminants 01/01/2000
Cormier, S M., E.L C. Lin, M. R. Millward, M SchubauerBerigan, D. E. Williams, B Subramanian, R. Sanders, B. Counts, AND D. Altfater. Using Regional Exposure Criteria and Upstream Reference Data to Characterize Spatial and Temporal Exposures to Chemical Contaminants. ENVIRONMENTAL TOXICOLOGY AND CHEMISTRY 19(4):1127-1135, (2000).
Abstract: Analyses of biomarkers in fish were used to evaluate exposures among locations and across time. Two types of references were used for comparison, an upstream reference sample remote from known point sources and regional exposure criteria derived from a baseline of fish from reference sites throughout Ohio, USA. Liver, bile and blood were sampled from white suckers (Catostomus commersoni) and common carp (Cyprinus carpio) collected during 1993 and 1996 in the Ottawa River near Lima, Ohio. Levels of exposure were measured for petroleum by naphthalene-type metabolites, combustion by-products by benzo(a)pyrene-type metabolites, coplanar organic compounds by ethoxyresorufin-0-deethylase (EROD) activity, and urea by blood urea nitrogen (BUN) levels. The four biomarkers analyzed proved effective in determining differences between reference and polluted sampling sites, between geographically close (<0.5 km) sites, and between sampling years at sites common in both years. Calculated exposure criteria levels of the polycyclic aromatic hydrocarbon bile metabolites were found to be a conservative approximation of levels from a designated reference site and could thereby permit comparison of biomarker levels of fish from the Ottawa River to a regional reference level. Polycyclic aromatic hydrocarbon bile metabolite and EROD activity levels were more reflective of spatial patterns of contamination than BUN, although all biomarkers indicated differences over time. Biomarkers from white suckers seemed to be more responsive in detecting changes in contaminanat levels than the same biomarkers from common carp. Lower levels in 1996 of all biomarkers at many sites suggested lower exposures than in 1993 and could be indicative of some improvement over the period.

JOURNAL Estimation of Exposure Criteria Values for Biliary Polycyclic Aromatic Hydrocarbon Metabolite Concentration in White Suckers (Catastomus Commersoni) 01/01/2000
Cormier, S M., E.L C. Lin, F A. Fulk, AND B Subramanian. Estimation of Exposure Criteria Values for Biliary Polycyclic Aromatic Hydrocarbon Metabolite Concentration in White Suckers (Catastomus Commersoni). ENVIRONMENTAL TOXICOLOGY AND CHEMISTRY. SETAC Press, Pensacola, FL, 19(4(2)):1120-1126, (2000).
Abstract: A methodology was developed for deriving quantitative exposure criteria useful for comparing a site or a watershed to a reference condition and for defining the occurrence of extreme exposures. The prototype method used indicators of exposures to oil contamination and combustion by-products, naphthalene (NAPH)-type and benzo(a)pyrene (BaP)-type biliary metabolites from white suckers (Catostomus commersoni). The fish were collected from first- to third-order streams in the Regional Environmental Monitoring Assessment Program (REMAP) study of the Eastern Corn Belt Plains ecoregion at randomly selected sites and from third- or higher-order streams in conjunction with the Ohio Environmental Protection Agency (OEPA) Biomonitoring Program, which sampled sites of concern and reference sites selected by ecologists. The polycyclic aromatic hydrocarbon metabolites were measured using fixed-wavelenth fluorescence with excitation/emission pairs at 290/335 nm for NAPH-type and 380/430 nm for BaP-type metabolites. Exposure criteria values were selected for each type of metabolite for both the REMAP study and the OEPA reference sites following the recommendations of the International Federation of Clinical Chemistry. Exposure criteria derived for each metabolite were not significantly different for REMAP and OEPA reference sites. More than one half of the OEPA nonreference sites were found to be contaminated, exceeding criteria values for both types of metabolites. This method for finding meaningful exposure criteria can be used to develop criteria of exposure to other contaminants for other wildlife and other ecosystems.

PRESENTATION A Sediment Toxicity Method Using Lemna Minor, Duckweed 11/14/2000
Lazorchak, J M., E. M. SuszcynskyMeister, AND M E. Smith. A Sediment Toxicity Method Using Lemna Minor, Duckweed. Presented at Society of Environmental Toxicology and Chemistry, Nashville, TN, November 12-16, 2000.
Abstract: We developed a Lemna minor sediment toxicity test method to assess sediment contaminants which may affect plants. This 96-hour test used 15 ml of sediment and 2 ml of overlying water which was renewed after 48 hours. Sand was used as the control sediment and also to dilute test sediments. This method was evaluated with sediments from the East River (ERS), NY; the NY/NJ harbor, the Calumet River, IL; and the Little Miami River, OH. In addition to the duckweek method, sediments were also tested using a miniaturized freshwater amphipod method and a fathead minnow embryo-larval (FHM) survival test. Of the three endpoints used in duckweed tests, results indicated that chlorophyll a was a more sensitive measure than either frond number or wet weight. ERS and NY/NJ sediments were toxic to H. azteca and FHM and inhibited L. minor growth. Tests were conducted using diluted NY/NJ harbor sediments, which were known to be uncontaminated by PAHs. Varying the overlying water volumes did not affect plant response to the control or test sediments. Nutrient enrichment of 1% NY/NJ sediment resulted in significantly higher levels of chlorophyll a as compared to unenriched sediments. Chlorophyll a measurements were as sensitive as survival of H. azteca to the Calumet sediments and more sensitive than the FHM survival. Sediments tested from the Little Miami River, which were shown to be inhibitory to H. azteca growth in earlier sediment tests, affected frond number and chlorophyll a content when tested with L. minor. A sediment reference toxicant method has been developed for KCl and Atrazine. The duckweed method is convenient to use and provides an additional sediment toxicity indicator.

PRESENTATION Hyperspectral Technique as An Indicator of Eutrophication and Sediment Load for Deep Rivers 11/14/2000
Cormier, S M., F A. Fulk, AND G. Senay. Hyperspectral Technique as An Indicator of Eutrophication and Sediment Load for Deep Rivers. Presented at Society of Environmental Toxicology and Chemistry, Nashville, TN, November 12-16, 2000.
Abstract: Remote sensing techniques were used to characterize and quantify spatial and temporal variation in water quality of the Great Miami River in Ohio. An initial feasibility study was conducted in the summer of 1999 using a non-imaging hand-held spectroradiometer to ascertain the presence of remotely detectable relationships between water quality parameters and spectral reflectance data. Ratios of narrow spectral bands in the red and infrared were significantly correlated with chlorophyll, turbidity and Secchi-disk depth readings. Spectral bands selected during the feasibility study were later used to program imaging sensors. Two sensors were flown aboard a small aircraft to acquire imagery in the Great Miami River basin. On 9/8-9/99, a CASI (Compact Airborne Spectrographic Imager) sensor acquired imagery in 19 different spectral bands at a spatial resolution of 2 m, and on 9/30/99, a HyMap sensor was flown with 126 spectral bands at a spatial resolution of 5 m. A total of about 80 km of the river was imaged duirng the first mission while a 20 km reach was covered on the second flyover to repeat coverage of the urban/industrial influences around the city of Dayton, Ohio. Instream measurements of water quality data such as turbidity, chlorophyll, nutrient concentrations and Secchi-disk depth were acquired within the same day of the overflights. Spectral indices were developed that showed promising correlation with groundtruthed water quality parameters such as algal chlorophyll, turbidity and Secchi disk depth. Maps of the relative distributions of chlorophyll and turbidity were created from the hyperspectral images of the river. In addition, spectral indices were developed that distinguish emergent vegetation from submerged vegetation.

PRESENTATION Miniaturized Sediment Procedures for Asessing Toxicity Using Marine and Freshwater Amphipods and Embryo/Larval Fish 11/14/2000
Lazorchak, J M. AND M E. Smith. Miniaturized Sediment Procedures for Asessing Toxicity Using Marine and Freshwater Amphipods and Embryo/Larval Fish. Presented at Society of Environmental Toxicology and Chemistry, Nashville, TN, November 12-16, 2000.
Abstract: Sediment toxicity tests are needed that can be conducted with less sediment volume and fewer organisms. Bench scale remediation techniques often produce less sediment than is required to perform the standardized sediment methods and the excess sediments that are generated present a potential hazardous waste disposal issue. Minimizing sample volumes reduces the time required to collect samples, prepare samples (sieving) and enumerate organisms at the end of the test. To address these issues we have modified existing USEPA methods that use significantly less sediment and fewer organisms than the standard toxicity tests and developed two alternative methods. Freshwater methods include a 7-day amphipod, Hyalella azteca method and 7-day fathead minnow (Pimephales promelas) embryo-larval hatching method and two marine methods, a 10-day amphipod, Ampelisca abdita, and a 7-day sheepshead minnow (Cyprinodon variegatus) embryo/larval method. We have modified the existing USEPA methods for the two amphipod tests and adapted the USEPA embryo/larval aqueous methods for sediment testing. In the freshwater amphipod test we have reduced the volume of sediment from 100 ml to 17 ml and reduced the number of organisms from 80 to 20. In the marine amphipod method we reduced the sediment volume from 175 ml to 40 ml and reduced the number of organisms from 100 to 50. Results on contaminated and uncontaminated sediments indicate that the miniaturized methods are comparable to the standardized test methods and the amount of time to prepare the samples and conduct the tests is significantly reduced. The miniaturized test methods can be used when regulatory program requirements are not required.

PRESENTATION Evaluation of Sampling Frequencies Required to Estimate Nutrient and Suspended Sediment Loads in Large Rivers 11/13/2000
Rowe, G. L. AND T Flum. Evaluation of Sampling Frequencies Required to Estimate Nutrient and Suspended Sediment Loads in Large Rivers. Presented at Society of Environmental Toxicology and Chemistry, Nashville, TN, November 12-16, 2000.
Abstract: Nutrients and suspended sediments in streams and large rivers are two major issues facing state and federal agencies. Accurate estimates of nutrient and sediment loads are needed to assess a variety of important water-quality issues including total maximum daily loads, aquatic ecosystem response and hypoxia in the Gulf of Mexico. Despite work by U.S. Geological Survey's National Water-Quality Assessment (NAWQA) program, questions remain concerning sampling frequency required to accurately estimate loads of nutrients and suspended sediments. A preliminary effort to determine appropriate sampling frequencies for estimating loads of nutrients and suspended sediments in a large river was made jointly by NERL and the Great and Little Miami River Basins NAWQA unit. Water samples were collected hourly with an automatic sampler at a single station on the Great Miami River just upstream from the confluence with the Ohio River. The GreAt Miami River was selected because available data indicate it has one of the highest yields of nutrients per unit area among watersheds contributing to the Gulf of Mexico. Hourly samples were composited into daily samples and analyzed for nitrate, ammonia, total phosphorus (TP), total Kjeldahl nitrogen (TKN) and suspended sediments (SS). For the period 9/99 to 4/00 ammonia concentrations ranged from 0.01 to 0.39 mg/L-1, nitrate from 0.10 to 8.35 mg/L-1, TP from 0.07 to 0.94 mg/L-1, TKN from 0.22 to 1.32 mg/L-1 and SS from 2.25 to 487 gm/L-1. Spring planting and feritlizer applications have begun and increased concentrations of nutrients and suspended sediment are expected. Analysis of availlable data indicates that loads estimated on the basis of monthly or bimonthly samples were as much as 30% less than loads estimated from daily composite samples.

PRESENTATION Macroinvertebrate Metrics and Water Chemistry Relationships from Non-Wadeable Streams of the Mid-Atlantic Integrity Assessment 11/13/2000
Klemm, D J., K A. Blocksom, W T. Thoeny, AND A. T. Herlihy. Macroinvertebrate Metrics and Water Chemistry Relationships from Non-Wadeable Streams of the Mid-Atlantic Integrity Assessment. Presented at Society of Environmental Toxicology and Chemistry, Nashville, TN, November 12-16, 2000.
Abstract: Macroinvertebrates were collected from non-wadeable streams during the summer of 1997 and 1998 for the USEPA Environmental Monitoring and Assessent Program (EMAP) Mid-Atlantic Integrated Assessment (MAIA). In this study we examined macroinvertebrate metrics to identify those which would help us assess biological condition in these streams. Because there were no designated reference or impaired sites we used water chemistry and minimum count criteria established for selecting reference sites among wadeable streams in the Mid-Atlantic Highlands region. We found distinct patterns in the data by examining metric values up to and beyond these criteria values. In addition we separated the 45 sites with water chemistry data into those fitting all criteria ("best" sites) , those fitting none ("worst" sites) and those in between. Using chemical-macroinvertebrate metric relationships and the ability of the metrics to discriminate the best and worst sites, we identified 5 out of 43 metrics evaluated which might be useful in assessing the biological integrity and health of the biological community in non-wadeable streams. These metrics included number of taxa, total number of individuals in the sample, % collector-gatherer individuals, number of Ephemeroptera, Trichoptera and Odonata taxa, and % tolerant taxa.

PRESENTATION Comparison of Mid-Western and Mid-Atlantic Regions Exposure Criteria for Petroleum and Combustion By-Products 11/13/2000
Cormier, S M., E.L C. Lin, B Subramanian, AND F A. Fulk. Comparison of Mid-Western and Mid-Atlantic Regions Exposure Criteria for Petroleum and Combustion By-Products. Presented at Society of Environmental Toxicology and Chemistry, Nashville, TN, November 12-16, 2000.
Abstract: Regional background levels of exposure to fish from petroleum and combustion by-products were determined for the state of Ohio (mid-Western) and the mid-Atlantic region. Exposures were measured using bile metabolites that fluoresce at 290/335 nm for naphthalene (NAPH)-type compounds and at 380/440 nm for benzo(a)pyrene (BAP)-type compounds. Fish were sampled using a probabalistic sampling design developed for the Environmental Monitoring Assessment Program (EMAP) in the mid-Atlantic and Eastern Cornbelt Plains Ecoregion (ECBP) and least impacted (LI) sites selected by Ohio EPA. Criteria for exposure values above background were determined using International Federation of Clinical Chemistry recommendations for reference values. Exposure criteria for NAPH for white suckers were: EMAP mid-Atlantic region 60 ug/mL/mg; ECBP 60 ug/mL/mg; and Ohio LI sites 80 ug/mL/mg. For BAP, exposure criteria for white suckers were: EMAP Mid-Atlantic region 0.4 ug/mL/mg; ECBP 0.4 ug/mL/mg; and Ohio LI sites 0.5 ug/mL/mg. These values were remarkably similar regardless of reigon or site selection. This suggests that a single exposure criteria value could be used to determine if fish have been exposed to petroleum or combustion by-products.

PRESENTATION Reference Condition Approach to the Assessment of Biological Integrity in Streams of the Southern Rocky Mountains and Its Use in Measuring the Effectiveness of Mile-Remediation Efforts 11/13/2000
Leland, H. V., A R. Selle, AND M. B. Griffith. Reference Condition Approach to the Assessment of Biological Integrity in Streams of the Southern Rocky Mountains and Its Use in Measuring the Effectiveness of Mile-Remediation Efforts. Presented at Society of Environmental Toxicology and Chemistry, Nashville, TN, November 12-16, 2000.
Abstract: A recent development in water quality assessment is the comparison of assemblage data from impacted stream segments with that for groups of segments representing reference (or minimally-impacted) conditions. The degree of impairment of a stream segment is expressed as metrics, such as the ratio of observed to expected (O/E) resident fauna. We examined the distribution of benthic invertebrates in streams of the southern Rocky Mountains physiographic province, Colorado, in relation to lithology, land cover, proximity to mines, water and sediment quality, and in-stream and riparian habitat. Our objective was to examine the potential for use of this method in monitoring the effectiveness of mine-remediation efforts based on data from approximately 50 reference segments and 85 moderately-impacted to impacted segments in the province. We used datasets from Regional EMAP and NAWQA studies conducted during the years 1994-1998. The results of this investigation are used to define requirements of a biomonitoring program to measure the effectiveness of mine-remediaton efforts in restoring biological resources of impacted streams and rivers of the Rocky Mountains.

PRESENTATION Bootstrapping and Monte Carlo Methods of Power Analysis Used to Establish Condition Categories for Biotic Indices 11/13/2000
Blocksom, K A., F A. Fulk, D J. Klemm, AND S M. Cormier. Bootstrapping and Monte Carlo Methods of Power Analysis Used to Establish Condition Categories for Biotic Indices. Presented at Society of Environmental Toxicology and Chemistry, Nashville, TN, November 12-16, 2000.
Abstract: Biotic indices have been used ot assess biological condition by dividing index scores into condition categories. Historically the number of categories has been based on professional judgement. Alternatively, statistical methods such as power analysis can be used to determine the number of categories distinguishable by a biotic index. Lacking the replication necessary to conduct traditional power analysis we used bootstrapping and Monte Carlo methods to conduct power analysis on two different biotic indices to establish condition categories. For the Stream Benthos Integrity Index (SBII) developed for wadeable streams in the Mid-Atlantic highlands we used several pairs of within year revisits to generate sets of bootstrap replicates. Using these samples we tested for differences between each possible pair of sites using t-tests. Repeating this process 500 times we determined the effect size (mean difference in scores) at which the power (proportion of tests for which the null hypothesis was rejected) was equal to 0.80. For the Lake Macroinvertebrate Integrity Index (LMII) developed for New Jersey lakes and reservoirs, we had replicate data for only 2 of 58 lakes. Thus we estimated sample variance from these replicates and used the maximum of the two variance values in our analysis. To generate samples for t-tests we used Monte Carlo methods to generate normal random variates based on a mean equal to the actual LMII score and variance equal to the maximum variance mentioned above. In each case the effect size was used to determine statistically the number of distinct condition categories for the index by dividing the maximum index score by the effect size.

PRESENTATION Region-Wide Genetic Structure of the Central Stoneroller (Campostoma Anomalum) and the Relationship of Genetic Diversity to Environmental Quality 11/13/2000
Franson, S E., M J. Bagley, S A. Christ, T K. Wessendarp, G P. Toth, AND M K. Smith. Region-Wide Genetic Structure of the Central Stoneroller (Campostoma Anomalum) and the Relationship of Genetic Diversity to Environmental Quality. Presented at Society of Enviornmental Toxicology and Chemistry, Nashville, TN, November 12-16, 2000.
Abstract: Anthropogenic stressors that reduce population size, alter migration corridors or modify mutational and selective forces on populations are expected to leave a lasting genetic footprint on the distribution of intraspecific genetic variation. Thus, the pattern of intraspecific genetic variation within and among populaitons should be a natural indicator of the condition of populations and the environments in which they are found. To the extent that extinction risk is determined by genetic variability, the pattern of genetic variation is also a useful indicator of population vulnerability. As a first step in gauging the usefulness of genetic diversity as an ecological indicator, we assessed the regional genetic structure of the cyprinid fish Campostoma anomatum (central stoneroller) in streams throughout the Eastern Cornbelt Plains Ecoregion. Patterns of genetic variation within and among populations were examined for samples collected from 110 sites using the random amplified polymorphic DNA (RAPD) technique. Sampling was done in conjunction with USEPA's Regional Environmental Monitoring and Assessment Program (REMAP), which provided simultaneous data for environmental variables that could be correlated to the genetic data. Analyses using Lynch's Similarity Index revealed that populations within the ecoregion were highly differentiated, indicating that gene flow between sites was generally low. Preliminary assessments of correlative environmental data suggest that genetic diversity of individuals within populations is positively correlated to both habitat quality (Qualitative Habitat Evaluation Index) and community structure (Index of Biotic Integrity). These preliminary resutls suggest that differences in genetic diversity among populations, when viewed in a regional context, are highly informative indicators of ecological condition and relative vulnerability of populations.

PRESENTATION Development of a Dna Archive for Genetic Monitoring of Fish Populations 11/13/2000
Bagley, M J., S E. Franson, T K. Wessendarp, S A. Christ, J M. Lazorchak, G P. Toth, AND M K. Smith. Development of a Dna Archive for Genetic Monitoring of Fish Populations. Presented at Society of Environmental Toxicology and Chemistry, Nashville, TN, November 12-16, 2000.
Abstract: Analysis of intraspecific genetic diversity provides a potentially powerful tool to estimate the impacts of environmental stressors on populations. Genetic responses of populations to novel stressors include dramatic shifts in genotype frequencies at loci under selection (i.e. adaptation) as well as large reductions in genetic diversity within populations at both neutral and selected loci (due to reduced effective population sizes). Typically inferences of genetic change are made through comparison of contemporaneous samples taken from spatially separated populations, some of which are treated as stressor-exposed "test" populations while others are treated as unexposed "reference" populations. In effect, these "genetic snapshots" are surrogates for more costly and logistically difficult temporal or spatio-temporal studies. The difficulty with the genetic snapshot approach is that the degree of genetic and ecological independence between test and reference populations, including the levels and types of past exposures, is rarely known with any accuracy. We believe that studies that directly measure genetic change within populations over time are critical to effective utilization of genetic markers and that the associated cost and logistical difficulties can be overcome if the proper infrastructure is provided. To aid in providing the necessary baseline, the USEPA created and maintains a DNA and fin-tissue archive for freshwater and estuarine fish. Access to the archive is open, and deposits by non-EPA investigators and institutions are encouraged. Current plans include creation of a web-accessible database of DNA collections cross-referenced to available phenotypic, enviornmental, and genetic data. Anticipated benefits of the archive include facilitation of some of the first studies designed to assess the evolutionary responses of populations to local, regional and global environmental change.

PRESENTATION Molecular Diagnostics Another Piece in the Environmental Puzzle 11/13/2000
Lattier, D L. AND G P. Toth. Molecular Diagnostics Another Piece in the Environmental Puzzle. Presented at Society of Environmental Toxicology and Chemistry, Nashville, TN, November 12-16, 2000.
Abstract: Molecular biology offers sensitive and expedient tools for the detection of exposure to environmental stressors. Molecular approaches provide the means for detection of the "first cellular event(s)" in response to environmental changes-specifically, immediate changes in gene expression. Environmental exposure monitoring using gene activity as an indicator is supported by the hypothesis that subcellular events resulting from an organism's contact with chemical milieus are manifested far in advance of those effects observed at higher levels of biological organization. Specifically this approach involves detection of changes in gene transcription and relative levels of tissue-specific messenger RNA (mRNA) which occur as a result of direct contact with xenobiotic chemicals present in the environment. Protein products that are synthesized in response to environmental change represent the terminal aspect in a multi-step biochemical pathway that is replete with diverse cellular control mechanisms. Most of these studies have centered on the single chemical/single gene response. This scheme largely ignores the reality of enviornmental complexity such as chemical fate and transport, synerigsm of chemical mixtures, multple genes competing for limited intracellular pools of transcription co-factors, and gene induction profiles resulting from chronically exposed organisms. Emerging technologies such as differential display and microarray DNA chips, provide a means to detect differences in inestimable gene products induced by the above scenarios. When applied judiciously and in concert with higher order ecological analyses, molecular biology can provide the important link between immediate environmental exposure and long term biological, community and population effects.

PRESENTATION Vitellogenin Gene Transcription as An Indicator of Exposure to 17-Alpha-Ethynylestradiol in Fathead Minnows 11/13/2000
Reddy, T V., D L. Lattier, J M. Lazorchak, S E. Franson, M E. Smith, E. M. SuszcynskyMeister, AND G P. Toth. Vitellogenin Gene Transcription as An Indicator of Exposure to 17-Alpha-Ethynylestradiol in Fathead Minnows. Presented at Society of Environmental Toxicology and Chemistry, Nashville, TN, November 12-16, 2000.
Abstract: Environmentally persistent chemicals that functionally mimic estrogen are ubiquitous in surface waters and have been shown to effect reproductive health of species living in these habitats. Toxicant induced transcription of specific genes is a sensitive indicator of exposure and directly correlates with an organism's biological condition in real time. Expression of the vitellogenin (Vg) genes, coding for the major egg yolk protein precursor in oviparous vertebrates, is hormonally regulated by estrogens and estrogen-like compounds. Using available DNA sequences for fathead minnow vitellogenin, we have designed synthetic oligonucleotide primers which detect VgmRNA in embryo/larvae and in the livers of adult fathead minnows in quantitative RT-PCR. Fathead minnow males, or larvae, were maintained in waterborne exposure conditions at various concentrations of 17-aplphpEthynylestradiol for 24 or 48 hrs. Vitellogenin mRNA, which is normally quiescent in males, was detected in both developmental stages, at concentrations as low as 0.0-25 mg/L. Agarose gel electrophoresis of RT-PCR products indicated cDNA, corresponding to Vg message, at the predicted molecular weight. This technology is being applied as an in situ exposure monitoring, using caged male fathead minnows, and in-laboratory testing of environmental water sources using adults and embryo-larvae. The use of the described monitoring scheme will enhance our knowledge of ecological exposures to estrogenic compounds across a range of surface waters.

PRESENTATION Productivity and Species Richness in Small Highland Streams: the Interaction of Habitat and Land Use History 06/15/2000
McCormick, F H. Productivity and Species Richness in Small Highland Streams: the Interaction of Habitat and Land Use History. Presented at American Society of Ichthyologists and Herpetologists, La Paz, B.C.S., Mexico, June 14-20, 2000.
Abstract: Small, upland, coldwater streams are an important resource for watershed management. In the Mid-Atlantic region, these streams are affected by acid deposition, mountaintop removal and valley fill for mineral extraction, and the effects of historical timber harvests. Small streams may be naturally unproductive, making the assessment of their condition based on fish species richness and biomass difficult to interpret. Fish assemblages in upland, coldwater systems were more depauperate, with fewer individuals, and represented by "pioneering" species such as blacknose dace (Rhinichthys atratulus) than valley and plateau streams of comparable size (50km2). Mean species richness, mean water column species richness, and mean number of trophic and reproductive guilds were significantly greater in lowland streams. However, mean number of sensitive species were greater in upland systems while the proportion of tolerant individuals were lower. Species richness variables (number of native species, families, species representing habitat preferences, or specific taxonomic groups) were higher at sites with abiotic characteristics (chemistry, land use, and habitat) indicative of human disturbance than for "reference" sites that had passed screens for those characteristics. While land use and its accompanying habitat degradation may account for low species richness in streams that would otherwise be expected to support greater native species diversity, differentiating impaired streams from naturally depauperate ones required evaluation on case by case, and often variable by variable, bases.

PRESENTATION Analysis of Lotic Macroinvertebrate Assemblages in California's Central Valley 05/30/2000
Griffith, M. B., P. Husby, R. K. Hall, P R. Kaufmann, AND B H. Hill. Analysis of Lotic Macroinvertebrate Assemblages in California's Central Valley. Presented at North American Benthological Society, Keystone, CO, May 29-June 2, 2000.
Abstract: Using multivariate and cluster analyses, we examined the relaitonships between chemical and physical characteristics and macroinvertebrate assemblages at sites sampled by R-EMAP in California's Central Valley. By contrasting results where community structure was summarized as metrics or genera abundances, we identified metrics or taxa diagnostic of lotic stressors and compared the sensitivities of these approaches to the stressors. Redundancy analysis (RDA) of metrics extracted three significant axes, accounting for 88% of the relation. RDA axes were correlated with channel morphology and substrates. Canonical correspondence analysis (CCA) of genera abudances extracted two significant axes, accounting for 23% of the relation. CCA axes were correlated with salts associated with irrigation and with channel morphology and substrates, but did not separate these gradients. Cluster analysis identified eight RDA groups and ten CCA groups. The differences among groups with respect to selected environmental variables were similar for metrics or genera abundances, but the agreement in group assignments based on the two approaches was not greater than expected if sites were assigned randomly. CCA measures variation primarily as the changes in genera abundances relative to each other. Metrics measure other characteristics, such as richness and taxa relative abudances. These approaches can be used together to design diagnostic metrics.

PRESENTATION Evaluation of Two Macroinvertebrate Sampling Methods for Non-Wadeable Streams 05/30/2000
Klemm, D J., K A. Blocksom, AND W T. Thoeny. Evaluation of Two Macroinvertebrate Sampling Methods for Non-Wadeable Streams. Presented at North American Benthological Society, Keystone, CO, May 29-June 2, 2000.
Abstract: Two macroinvertebrate sampling protocols for non-wadeable streams were evaluated during the summer of 1997 and 1998 as part of the USEPA's Environmental Monitoring and Assessment Program (EMAP), Mid-Atlantic Integrated Assessment (MAIA). Macroinvertebrates were collected by kick net and drift net methods to determine if these methods would adequately collect 300 organisms (or within 10% of 300) that could be used for bioassessment of a non-wadeable stream segment. Eleven transects were marked off equidistant in proportion to stream size, and the benthic macroinvertebrate assemblages collected on one side of the stream. In 1997, one kick net sample was collected from each of 11 transects and all samples composited, but in 1998 two kick net samples were collected from each of 11 transects. For both years, two drift net samples were collected and composited per site. In 1997 and 1998, kick net samples and drift net samples were collected from 13 and 34 non-wadeable stream sites, respectively. Using EMAP laboratory protocols for processing of benthic invertebrate samples, the kick net samples were split into thirds, and the random split sub-samples were randomly sorted using a tray containing 40, 5 cm by 5 cm squares until 300 organisms were picked, and all sub-samples sorted if necessary. The drift net samples were not split, and the entire sample was randomly picked. The number of individuals collected with the drift net and kick net were compared across years and within each year. In 1997, neither sampling device collected 300 or more organisms (the performance criteria), while in 1998, 27% of kick net samples (9 of 34) and 3% of drift net samples (1 of 34) collected 300 or more organisms. As expected, the median number of taxa and individuals for kick net samples in 1998 (47 and 279, respectively) were much higher than for 1997 (23 and 62, respectively). In addition, the drift nets collected consistently much fewer taxa and individuals than kick nets in both years (medians 18 and 44, respectively). It is recommended that three kick net samples be collected at each transect and that split sub-samples be continually picked until the number of organisms picked from the splits are within 10% of 300. It is also reocmmended that the drift net sampling method be dropped from the EMAP non-wadeable stream protocols because the results indicate poor sampling power to collect benthic macroinvertebrates during the time of day the drift nets are employed.

PRESENTATION Diatom Species Richness in Streams of the Eastern US: Stream Size and Habitat Effects 05/29/2000
Hill, B H., S DeCelles, AND M. Vander Borgh. Diatom Species Richness in Streams of the Eastern US: Stream Size and Habitat Effects. Presented at North American Benthological Society, Keystone, CO, May 29-June 2, 2000.
Abstract: We analyzed the relationship between benthic diatom assemblages, stream size, and habitat characteristics in 445 first through seventh order streams in the Mid-Atlantic (n=230), South Atlantic (n=61), Ohio (n=140), and Tennessee (n=14) hydrologic regions. Diatom samples were collected from 11 transect locations in each stream and combined to yield a single sample per site. Sampling was done as a part of the USEPA's Environmental Monitoring and Assessment Program (EMAP) surveys in 1997 and 1998. Nearly 600 species of diatoms were collected, though species richness at any one site never exceeded 70. Diatom richness per site was greatest, on average, in streams of the South Atlantic region (40 species), followed by those of the Mid-Atlantic (32 spp.), Ohio (32 spp.), and Tennessee (28 spp.) regions. Stream order was a weak predictor of species richness, accounting for only 3-13% of the variance in the data by hydrologic region. The addition of in situ stream size and habitat measurements improved model r2 to 0.27-0.66. We used canonical correlation analysis to investigate the relationship between species richness and environmental gradients. Richness was significantly correlated with canonical axes representing stream size and in-stream substrate size.

PRESENTATION Development and Evaluation of a Macroinvertebrate Benthos Integrity Index (Sbii) for Bioassessment of Streams in the Mid-Atlantic Highlands Region 05/29/2000
Klemm, D J., F A. Fulk, S M. Cormier, K A. Blocksom, AND W T. Thoeny. Development and Evaluation of a Macroinvertebrate Benthos Integrity Index (Sbii) for Bioassessment of Streams in the Mid-Atlantic Highlands Region. Presented at North American Benthological Society, Keystone, CO, May 29-June 2, 2000.
Abstract: A multimetric macroinvertebrate index called the Stream Benthos Integrity Index (SBII) was developed for the Mid-Atlantic Highlands Region of the United States. The SBII was developed for assessing biological conditions of wadeable streams and was based on benthic macroinvertebrate assemblage data that were collected during 1993-1995 as part of the USEPA's Environmental Monitoring and Assessment Program (EMAP) and the Mid-Atlantic Highlands Assessment (MAHA). Macroinvertebrates were collected from riffle/run habitats of first, second, and third order wadeable streams and identified to the lowest taxon possible in the laboratory. The responses of macroinvertebrate metrics to physical habitat and chemical indicators of catchment stream disturbance for 407 stream sites were used and analyzed for this study. Forty-two candidate macroinvertebrate metrics were evaluated statistically using a stepwise process that eliminated metrics not meeting specific criteria. Each metric was evaluated for 1) discriminatory power, 2) scope of impairment, 3) redundancy, 4) relationships to stressors, and 5) repeatability. The relationships among macroinvertebrate metrics, habitat integrity, anthropogenic disturbance and knowledge of reference and impacted sites based on chemical and physical habitat criteria were used to develop the SBII for the assessment of stream condition in the Mid-Atlantic Highlands Region. The SBII consists of 8 metrics which include population, community and pollution tolerance metrics (Number of taxa, HBI, % intolerant taxa, % non-insect individuals, % Chironomid taxa, % Plecoptera taxa, EPA Taxa Index, and % Oligochaete/Leech Individuals), chosen using statistical criteria and best professional judgment for assessing anthropogenic stressors. The stream condition based on the macroinvertebrate population and community data of riffle/run habitats was assessed using the SBII. Results indicate that the SBII will improve the ability of USEPA regions, states, municipalities, and industry to determine the biological integrity and conditions of freshwater wadeable streams and to improve their understanding, protection, preservation, and restoration of stream ecosystems.

PRESENTATION Development of An Index of Benthos Integrity Using Macroinvertebrates for New Jersey Lakes and Reservoirs 05/29/2000
Blocksom, K A., J. P. Kurtenbach, D J. Klemm, F A. Fulk, AND S M. Cormier. Development of An Index of Benthos Integrity Using Macroinvertebrates for New Jersey Lakes and Reservoirs. Presented at North American Benthological Society, Keystone, CO, May 29-June 2, 2000.
Abstract: Recently there has been a greater focus by the USEPA on bioassessment and biocriteria development for lakes and reservoirs. In this study a multimetric index was developed to evaluate the condition of New Jersey lakes and reservoirs using benthic macroinvertebrates. Samples were collected from 58 New Jersey lakes during summer 1997. Thirty-three candidate metrics were evaluated using a step-wise process to evaluate discriminatory power, relative scope of impairment, redundancy, and stressor relationships with metrics, eliminating those failing to meet specific criteria. Five metrics were selected for the Lakes Macroinvertebrate Integrity Index (LMII): Hilsenhoff Biotic Index, percent chronomid individuals, percent collector-gatherer taxa, percent oligochaetes and leeches, and number of Diptera taxa. Scores were based on the cumulative distribution function of each metric, and metric scores represent the fraction of the best attainable value (95th or 5th percentile) achieved at a site. The five metric scores were summed into the LMII. The LMII discriminates well between reference and impaired lakes and is strongly related to several stressors. Biocriteria were created based on the distribution of LMII scores among reference lakes. The results suggest that lakes in more heavily populated areas are more likely to be in poor condition.

PRESENTATION How to Distinguish Chemical and Physical Causes of Biological Impairment: Example from a Stream in the United States 05/21/2000
Cormier, S M., E.L C. Lin, AND B Subramanian. How to Distinguish Chemical and Physical Causes of Biological Impairment: Example from a Stream in the United States. Presented at Society for Environmental Toxicology and Chemistry, London, England, May 21-25, 2000.
Abstract: Attributes of fish and invertebrate assemblages in rivers and streams can be altered by either physical, biological or chemical causes. In order to determine the active cause in specific situations, certain types of ecological data can be used to eliminate potential causes. By deduction, the probable causes can be determined and experimentally confirmed. Multiple lines of evidence were used in a logical sequence to identify causes of impairment in a stream in Ohio that was potentially impacted by toxic chemicals, low dissolved oxygen, channel modification, nutrient enrichment, or a combination of these factors. In some segments, channelization was a critical factor, and in others, toxic chemicals. Overall, the process was useful for presenting a highly defensible case for attributing causes of biological impairment that resulted in a plan to remediate the stream.

PRESENTATION USEPA Stressor Identification Guidance 04/25/2000
ReedJudkins, D., S M. Cormier, G Suter, S B. Norton, W J. Mitchell, AND B. Swietlik. USEPA Stressor Identification Guidance. Presented at National Water Quality Monitoring Council, Austin, TX, April 25-27, 2000.
Abstract: Biological assessments integrate the effects of multiple stressors (chemical, biological and physical) on biological communities. For a variety of reasons, difficulties often arise in identifying and prioritizing stressors that are likely causative agents. New guidance is in preparation by USEPA to assist resource managers in identifying stressors to aquatic biota. Future needs are being evaluated and research plans developed to assist in accurate stressor identification. The strategy will be presented.

PRESENTATION Evaluation of Genetic Damage in Fish Exposed to Pesticides in Field Aquatic Microcosms 04/09/2000
Chang, L W., J R. Meier, G P. Toth, AND D. W. Graham. Evaluation of Genetic Damage in Fish Exposed to Pesticides in Field Aquatic Microcosms. Presented at Environmental Mutagen Society, New Orleans, LA, April 8-13, 2000.
Abstract: Single cell gel electrophoresis (SCG) and micronucleus (MN) assays were used to measure DNA strand breaks and chromosomal damage in fish blood erythrocytes as biological indicators of exposure to alachlor and atrazine in a surrogate aquatic ecosystem. Caged common carp (Cyprinus carpio) were exposed to either 0, 10, 25, 50 and 100 ug/L alachlor for 60 days or 0, 3, 10, 30 and 100 ug/L atrazine for 30 days in field microcosms. Benzo(a)pyrene (BaP), 0.76 ug/L) was used as a positive control. Blood samples from 6 fish of each treatment group were taken by caudal puncture at 0, 1, 4, 30 and 60 days after exposure. For the SCG assay, the overall mean values of tail length, tail moment and % tail DNA were analyzed by computerized image analysis for the extent of DNA damage. For the MN assay, blood smears were stained with acridine orange. The ratio of polychromatic erythrocytes (PCE) to normochromatic erythrocyte (NCE) and MN frequencies were scored manually by fluorescence microscopy. The results of the SCG assay indicate alachlor induced statistically significant dose-related DNA damage in carp blood erythrocytes as shown by three parameters during the entire exposure period. The level of damage reached a peak at 50-100 ug/L. For atrazine, a maximal response was seen at a concentration of 10 ug/L, followed by a plateau of the dose-response curve at 30 and 100 ug/L. BaP induced significant DNA damage at 0.76 ug/L. For all three chemicals, significant increases in damage were seen by one day after exposure, with the peak response being at day 4, followed by a slight decline in response at day 30 or 50. As for the MN assay, the results show the PCE frequencies were very low on most of the carp blood smears of both control and exposed fish. The PCE level decreased from day 4 to day 30 of the study duration. No MN were found in any PCE or NCE in any of the fish including those exposed to BaP. These surrogate ecosystem studies demonstrate the usefulness of the SCG assay for monitoring chemical stressors with genotoxic activity in aquatic ecosystems. The negative MN results are inconclusive because of the low erythrocyte turnover in the fish.

PRESENTATION Indicators of Genetic Damage in Model Stream Fish Using Controlled Laboratory Exposures 04/08/2000
Meier, J R., L W. Chang, P A. Wernsing, AND S. Gibbs. Indicators of Genetic Damage in Model Stream Fish Using Controlled Laboratory Exposures. Presented at Environmental Mutagen Society, New Orleans, LA, April 8-13, 2000.
Abstract: The micronucleus (MN) and single cell gel electrophoresis (SCG) assays are being applied to peripheral blood cells from several fish species using model genotoxicants and envirornmentally relevant pollutants. In initial studies, bluegill sunfish (Lepomis macrochirus, 4 to 5 per treatment group) were exposed by i.p. injection to 4-nitroquinoline n-oxide (NQO) at doses of 0, 7.5, 15 and 30 mg/kg or to mitomycin C (MMC) at doses of 0, 1 and 2 mg/kg. Blood samples were taken by caudal puncture at 1 and 3 days after exposure for NQO and at 1, 2, 4, and 8 days for MMC. For the SCG assay, the tail length, tail moment and 5 tail DNA parameters were analyzed by computerized image analysis. For MN assay, acridine orange-stained blood smears were scored manually for micronuclei in polychromatic erythrocytes (PCE) and normochromatic erythorcytes (NCE). Control MN frequencies at all time points were very low (mean = 0.08 MN/1,000 PCEs). No MN were induced at 1 day after exposure to either chemical. By 3 days, a significant elevation in MNPCEs was seen at all thee NQO doses. Significant increases in MN were also observed with MMC, with a peak response of 15 MN/1,000 PCEs at 4-8 days after treatment. For both chemicals, time- and dose-dependent toxicity to the erythropoietic system was observed, based on a decreased PCE:NCE ratio. Evaluation of DNA damage using the SCG assay indicated a significant, dose-related response at 1 day for both NQO and MMC. By 3 days after NQO treatment, the level of DNA damage declined slightly, and for MMC no significant responses were observed at 2-8 days after treatment. These results indicate that monitoring SCG and MN endpoints in fish erythrocytes may provide complementary indicators of exposure of differing duration to genotoxic environmental contaminants.

PRESENTATION Toxicity Evaluation of 1,3,5-Trinitrobenzene (Tnb) in Shrews (Cryptotis Parva) 03/17/2000
Reddy, T V., F B. Daniel, G. R. Olson, B E. Wiechman, AND G. Reddy. Toxicity Evaluation of 1,3,5-Trinitrobenzene (Tnb) in Shrews (Cryptotis Parva). Presented at Society of Toxicology, Philadelphia, PA, March 17-24, 2000.
Abstract: TNB has been detected as an environmental contaminant of soil and water at certain Army installations and production waste disposal sites. At present there are no toxicity data on TNB in small wild mammals that might be used for ecological risk assessment. Therefore, a 14-day toxicity study was conducted to determine the No Observed Adverse Effect Level (NOAEL) of TNB in the least shrew (Cryptotis parva). Male and female shrews (10/group/sex) were fed a mixture of wet-dry Purina Cat Chow containing 0, 5, 10, 20 and 40 pm TNB for 14 days. The calculated average consumed doses were 0, 10.68, 22.24, 37l79 and 98.27 mg/kg/day for males and 0, 10.75, 21.60, 45.26 and 98.72 mg/kg/day for females. There were no significant differences in food and water consumption among the groups. However, there was a decrease in body weight in males (10 ot 40 ppm) and a significant increase in body weight in females in the 20 ppm dose group. The relative spleen weights were increased in females (10 to 40 ppm). There were no significant changes in hematology parameters in females, while in males there was a significant decrease in RBC and an increase in WBC in the 40 ppm dose group. One histopathological change was erythroid cell hyperplasia in the spleen of males receivig 20 and 40 ppm TNB diet. The NOAEL for male and female shrews was determined as 10.75 mg TNB kg/day for the 14-day study.

PRESENTATION Determining Background Exposure to Petroleum and Combustion By-Products: Comparison of Mid-Western and Mid-Atlantic Regions 01/18/2000
Cormier, S M., E.L C. Lin, B Subramanian, AND F A. Fulk. Determining Background Exposure to Petroleum and Combustion By-Products: Comparison of Mid-Western and Mid-Atlantic Regions. Presented at National Science Meeting, Toronto, Canada, January 18-20, 2000.
Abstract: Regional background levels of exposure to fish from petroleum and combustion by-products were determined for the state of Ohio and the mid-Atlantic region. Exposures were measured using bile metabolites that fluoresce at 290/335 nm for naphthalene(NAPH)-type compounds and at 380/440 nm for benzo(a)pyrene(BAP)-type compounds. Fish were sampled using a probabalistic sampling design developed for the Environmental Monitoring Assessment Program in the mid-Atlantic and Eastern Cornbelt Plains Ecoregion and least impacted (LI) sites selected by Ohio EPA. Criteria for exposure values above background were determined using International Federation of Clinical Chemistry (IFCC) recommendations for reference values. Exposure criteria for NAPH for white suckers were: EMAP MAIA 60 ug/mL/mg; Ohio REMAP 60 ug/mL/mg; and Ohio LI sites 80 ug/mL/mg. For BAP, exposure criteria for white suckers were: EMAP MAIA 0.4 ug/mL/mg/ Ohio REMAP 0.4 ug/mL/mg; and Ohio LI sites 0.5 ug/mL/mg. These values were remarkably similar regardless of region or site selection. This suggests that a single exposure criteria value could be used to determine if fish have been exposed to petroleum or combustion by-products.

PRESENTATION Development of Benthic Macroinvertebrate Index for Measuring the Condition of Streams at a Regional Scale 01/18/2000
Cormier, S M., D J. Klemm, AND F A. Fulk. Development of Benthic Macroinvertebrate Index for Measuring the Condition of Streams at a Regional Scale. Presented at National Science Meeting, Toronto, Canada, January 18-20, 2000.
Abstract: A multimetric macroinvertebrate index of stream condition was developed for the Mid-Atlantic Highlands Region of the United States. Benthic macroinvertebrate samples were collected from 562 first through third order streams between 1993 and 1995. Macroinvertebrates were collected from riffles or runs and identified to the lowest possible taxon (genus and species). Measurements of physical habitat and water chemistry were collected at 407 stream sites. Forty-two candidate metrics were evaluated in a logical, statistical process that eliminated metrics using three criteria: 1) each metric's ability to discriminate sites with better chemical and physical habitat indicators from impaired sites; 2) each metric's independence or lack of redundancy with other metrics; and 3) each metric's relationship with stressors. Final selection of eight metrics was based on best performance for redundant (autocorrelated) metrics, ecological relevance, acceptance by scientific community, and added value to the performance of the complete index. The final Stream Benthos Invertebrate Index (SBII) consisted of eight metrics: total number of taxa, number of EPT taxa, % intolerant taxa, % non-insect individuals, % chironomid taxa, % plecoptera taxa, % oligochaete/leech individuals, and the Hilsenhoff Biotic Index (HBI). When the index was compared with the first axis from a principal component analysis using the original forty-two metrics, the correlation was highly significant. The process that was used to develop the index provides confidence because it is based on a jargon, logical process that is consistent with ecological theory using metrics that are accepted by state biologists. Furthermore, there is independent evidence of ability to detect impairment and independent confirmation from multivariate analysis. Thus, comparisons of biological conditions in streams can be confidently reported across regional scales.

PRESENTATION Indicator of Eutrophication and Sediment Load: Hyperspectral Technique for Continuous Coverage of Deep Rivers 01/18/2000
Cormier, S M., F A. Fulk, AND G. Senay. Indicator of Eutrophication and Sediment Load: Hyperspectral Technique for Continuous Coverage of Deep Rivers. Presented at National Science Meeting, Toronto, Canada, January 18-20, 2000.
Abstract: Remote sensing techniques were used to characterize and quantify spatial and temporal variation in water quality of the Great Miami River in Ohio. An initial feasibility study was conducted in the summer of 1999 using a non-imaging hand-held spectroradiometer to ascertain the presence of remotely detectable relationships between water quality parameters and spectral reflectance data. Ratios of narrow spectral bands in the red and infrared were significantly correlated with chlorophyll, turbidity and secchi-diskdepth readings. Spectral bands selected during the feasiblity study were later used to program the imaging sensors. Two sensors were flown aboard a small aircraft to acquire imagery in the Great Miami River basin. On September 8 and 9, 1999, a CASI (Compact Airborne Spectrographic Imager) sensor acquired imagery in 19 different spectral bands at a spatial resolution of 2 m, and on September 30, 1999, a HyMap sensor was flown with 126 spectral bands at a spatial resolution of 5 m. A total of about 80km of the river was imaged during the first mission while a 20 km reach was covered on the second flyover to repeat coverage of the urban/industrial influences around the city of Dayton, Ohio. Instream measurements were acquired within the same day of the overflights. Water quality data such as turbidity, chlorophyll, nutrient concentrations (nitrogen and phosphorous) were measured to calibrate the image. Currently, spectral processing of the imagery is underway to differentiate, quantify and map the distribution of algal chlorophyll, sediment loads and aquatic vegetation.

PUBLISHED REPORT Field Operations and Methods for Measuring the Ecological Condition of Non-Wadeable Rivers and Streams 09/30/2000
Lazorchak, J M., B H. Hill, D. K. Averill, D V. Peck, AND D J. Klemm. Field Operations and Methods for Measuring the Ecological Condition of Non-Wadeable Rivers and Streams. U.S. Environmental Protection Agency, Washington, DC, EPA/620/R-00/007, 2000.
Abstract: The methods and instructions for field operations presented in this manual for surveys of non-wadeable streams and rivers were developed and tested based on 55 sample sites in the Mid-Atlantic region and 53 sites in an Oregon study during two years of pilot and demonstration projects (1997 and 1998). These projects were conducted under the sponsorship of the U.S. Environmental Protection Agency and its collaborators through the Environmental Monitoring and Assessment Program (EMAP). The program focuses on evaluating ecological conditions on regional and national scales. This document describes procedures for collecting data, samples and information about biotic assemblages, environmental measures or attributes of indicators of non-wadeable stream ecosystem condition. The procedures presented in this manual were developed based on standard or accepted methods, modified as necessary to adapt them to EMAP sampling requirements. They are intended for use in field studies sponsored by EMAP. In addition to methodology, additional information on data management, safety and health and logistical aspects is integrated into the procedures and overall operational scenario. Procedures are described for collecting field measurement data and/or acceptable index samples for several response and stressor indicators, including water chemistry, physical habitat, benthic macroinvertebrate assemblages, aquatic vertebrate assemblages, fish tissue contaminants, periphyton assemblages, and sediment community metabolism. The manual described field implementation of these methods and the logistical foundation constructed during field projects. Flowcharts and other graphic aids provide overall summaries of specific field activities required to visit a river site and collect data for these indicators. Tables give step-by-step protocol instructions. These figures and tables can be extracted and bound separately to make a convenient quick field reference for field teams. The manual also includes example field data forms for recording measurements and observations made in the field and sample tracking information. Checklists of all supplies and equipment needed for each field task are included to help ensure that these materials are available when required.

 

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