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Ecological Exposure Research Division Publications: 2004

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This page lists publication titles, citations and abstracts produced by NERL's Ecological Exposure Research Division for the year 2004, organized by Publication Type. Your search has returned 66 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
BOOK CHAPTER A Toxicity Assessment Approach for Evaluation of in-Situ Bioremediation of PAH Contaminated Sediments 12/01/2004
Tabak, H H., J M. Lazorchak, M E. Smith, AND J. A. Ferretti. A Toxicity Assessment Approach for Evaluation of in-Situ Bioremediation of PAH Contaminated Sediments. Chapter 27, Dr. Gary Ostrander (ed.), Techniques in Aquatic Toxicology. CRC Press LLC, Boca Raton, FL, 2:479-509, (2004).
Abstract: Polycyclic aromatic hydrocarbons (PAHs) represent a group of organic contaminants known for their prevalence and persistence in petroleum-impacted environment such as groundwater, soils and sediments. Many high molecular weight PAHs are suspected carcinogens and the existence of these PAHs in dredged sediments can result in the classification of the sediment as hazardous waste, for which appropriate treatments may be necessry prior to disposal. Unfortunately ex-situ PAH treatment is not cost effective and natural attenuation/recovery has uncertainties including possible resuspension, transport and distribution over wide areas, that can produce substantial risk to human health and to the ecosystems. The widespread contamination of sediments by PAHs has thus created a need for a cost-effective remediation process for restoration of these sediments and bioremediation is one method that may reduce the risk of sediment associated PAHs.

INTERNAL REPORT A Compendium of Chemical, Physical and Biological Methods for Assessing and Monitoring the Remediation of Contaminated Sediment Sites 05/21/2004
Lazorchak, J M. AND J. Josephs. A Compendium of Chemical, Physical and Biological Methods for Assessing and Monitoring the Remediation of Contaminated Sediment Sites. U.S. Environmental Protection Agency, Washington, DC, EPA/600/R-03/108.
Abstract: Considering the many organizations which have published methods for monitoring contaminated sediments and the large number of documents on this subject, it can be a formidable task for a superfund project manager to find methods appropriate for his or her contaminated sediment site. This compendium of chemical, physical and biological methods for assessing and monitoring the remediation of contaminated sediment sites has been prepared to inform superfund project managers and others about appropriate methods for monitoring and assessing the remediation of contaminated sediments. Although the document can be printed as a text document, it is also intended to be viewed on a computer screen in order to take advantage of its hypertext links to navigate the document and to access reference documents available on the internet. Search engines can also be utilized to locate information contained in the document. The methods included focus primarily on published or otherwise citeable chemical, physical and biological testing methodologies used by EPA at superfund sites. The document summarized the methods, including references to the methods and hypertext links to access those methods which are available on the internet. Without exception, it is intended that all of the methods presented will be suitable for investigations at superfund sites containing contaminated sediments. However, not all methods will be suitable for all sites. The selection of methods for a particular site will depend on the site conditions, remediation plans, budgetary constraints and other factors.

JOURNAL Evaluation of the Model Anti-Androgen Flutamide for Assessing the Mechanistic Basis of Responses to An Androgen in the Fathead Minnow (Journal Article) 12/03/2004
ANKLEY, G. T., D. L. DEFOE, M. D. KAHL, K. M. JENSEN, E. A. MAKYNEN, A. L. MIRACLE, P. C. HARTIG, V. S. WILSON, L. E. GRAY, AND M. C. CARDON. Evaluation of the Model Anti-Androgen Flutamide for Assessing the Mechanistic Basis of Responses to An Androgen in the Fathead Minnow (Journal Article). ENVIRONMENTAL SCIENCE AND TECHNOLOGY. John Wiley & Sons, Ltd., Indianapolis, IN, 38(23):6322-6327, (2004).
Abstract: In this study we characterized the effects of flutamide, a model mammalian androgen receptor (AR) antagonist, on endocrine function in the fathead minnow (Pimephales promelas), a small fish species which is widely used for testing endocrine-disrupting chemicals (EDCs). Binding assays with whole cells transiently-transfected with cloned fathead minnow AR indicated that flutamide binds competitively to the receptor. However, as is true in mammalian systems, a 2-hydroxylated metabolite of flutamide binds to the AR with a much higher affinity than the parent chemical. Mixture experiments with flutamide and the androgen 172-trenbolone demonstrated that the anti-androgen effectively blocked trenbolone-induced masculinization (nuptial tubercle production) of female fathead minnows, indicating antagonism of an AR receptor-mediated response in vivo. Conversely, reductions in vitellogenin in trenbolone-exposed females were not blocked by flutamide, suggesting that the vitellogenin response is not directly mediated through the AR. The results of these studies provide data demonstrating the validity of using the fathead minnow as a model species for detecting EDCs that exert toxicity through interactions with the AR.

JOURNAL Dispersal and Mortality of Red-Shouldered Hawks Banded in Ohio 11/01/2004
Dykstra, C. R., J. L. Hays, M. M. Simon, J. B. Holt, G. R. Austing, AND F B. Daniel. Dispersal and Mortality of Red-Shouldered Hawks Banded in Ohio. JOURNAL OF RAPTOR RESEARCH 38:304-311, (2004).
Abstract: We banded nestling red-shouldered hawks (Buteo lineatus) in southwestern Ohio and northern Kentucky (SW OHIO, hereafter) to examine movements and determine causes of mortality in this suburban population. For comparison, we examined band recovery records for nestling red-shouldered hawks banded in rural northern Ohio. Of 899 nestlings banded in SW OHIO between 1995 and 2002, 43 (4.8%) were encountered (dead or alive) sometime after fledging. Mean distance from natal nest at time of encounter was 38.5 +/- 13.6 km and was not correlated with hawk age (P>0.58). Distance from natal nest did not differ for hawks of three age classes or between those encountered in the breeding or non-breeding seasons (P>0.13). Cumulative exponential distribution (CED) analysis of distance from natal nest at time of encounter indicated that 50% of SW OHIO red-shouldered hawks were found <15 km from their natal nest, 75% <29 km away and 95% <62 km away. Mean age of hawks recovered dead was 1.9 +/- 0.4 years (N=31). CED analysis of age at recovery indicated that 50% of red-shouldered hawks were dead by age 1.2 years, 75% by 2.4 years, and 95% by 5.2 years. SW OHIO hawks did not differ from hawks banded in northern Ohio in either distance from natal nest or age at recovery.

JOURNAL Characterization of Seven Polymorphic Microsatellite Loci in the Common Loon (Gavia Immer) 06/01/2004
McMillan, A., M J. Bagley, AND D. Evers. Characterization of Seven Polymorphic Microsatellite Loci in the Common Loon (Gavia Immer). MOLECULAR ECOLOGY NOTES. Blackwell Publishing Ltd, 4(2):297-299, (2004).
Abstract: We describe polymerase chain reaction (PCR) primers and conditions to amplify seven microsatellite DNA loci isolated from the Common Loon (Gavia immer). The PCR primers were tested on 83 individuals from ten locations in North America, including breeding, migration stopover, and wintering areas. Between two and seven alleles were observed to segregate at the seven microsatellite loci, with observed heterozygosities ranging from 0.048 to 0.695.

JOURNAL Temporal and Spatial Variability in the Estrogenicity of a Municipal Wastewater Effluent 03/01/2004
Hemming, J. M., H. J. Allen, K. A. Thuesen, P. K. Turner, W. T. Waller, J M. Lazorchak, D L. Lattier, M. Chow, N Denslow, AND B. Venables. Temporal and Spatial Variability in the Estrogenicity of a Municipal Wastewater Effluent. ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY 57(3):303-310, (2004).
Abstract: Estrogenicity of a municipal wastewater effluent was monitored using the vitellogenin biomarker in adult male fathead minnows (Pimephales promelas). Variability in expression of the vitellogenin biomarker was evident among monitoring periods. Significant increases in plasma vitellogenin concentrations were detected in the months of March and December, but not in August or June. Additionally, the magnitude of expression was variable. Variability in the spatial scale was also evident during the March and June exposure months. Concurrent exposures in both the creek receiving the effluent from a wastewater treatment plant and an experiemntal wetland showed estrogenicity to be different with distance from the respective effluent inflow sites. March exposures showed estrogenicity to be somewhat persistent in the receiving creek, but to decrease rapidly within the experimental wetland. Results will be discussed relative to monitoring season, spatial distribution of response in both receiving systems, and relative to possible causative factors contributing to the effluent estrogenicity.

JOURNAL Journal Article: Analysis of in Vivo and in Vitro Dna Strand Breaks from Trihalomethane Exposure 02/17/2004
Geter, D. R., L W. Chang, N. M. Hanley, M. K. Ross, R. A. Pegram, AND A B. DeAngelo. Journal Article: Analysis of in Vivo and in Vitro Dna Strand Breaks from Trihalomethane Exposure. CARCINOGENESIS 3(1):2, (2004).
Abstract: Epidemiological studies have linked the consumption of chlorinated surface waters to an increased risk of two major causes of human mortality, colorectal and bladder cancer. Trihalomethanes (THMs) are by-products formed when chlorine is used to disinfect drinking water. The purpose of this study was to examine the ability of the THMs, trichloromethane (TCM), bromodichloromethane (BDCM), dibromochloromethane (DBCM), and tribromomethane (TBM) to induce DNA strand breaks (SB) in (1) CCRF-CEM human lymphoblastic leukemia cells, (2) primary rat hepatocytes (PRH) exposed in vitro, and (3) rats exposed by gavage or drinking water.

PRESENTATION 2002/2003 National Screening Survey of Edcs in Municipal Wastewater Treatment Facilities 12/02/2004
LAZORCHAK, J. M., M. MILLS, G. D. SAYLES, R. C. BRENNER, A. D. BIALES, AND G. P. TOTH. 2002/2003 National Screening Survey of Edcs in Municipal Wastewater Treatment Facilities. Presented at ORSANCO Research Committee Meeting, Cincinnati, OH, December 02, 2004.
Abstract: Oral presentation at the meeting of the ORSANCO Research Committee, December 2, 2004.

PRESENTATION Assessing the Use of Oligonucleotide Microarrays for Fathead Minnow (Pimephales Promelas) to Examine Exposure Variables 11/15/2004
Miracle, A L., I Knoebl, D Villeneuve, G Ankley, N Denslow, B. Carter, AND P. Larkin. Assessing the Use of Oligonucleotide Microarrays for Fathead Minnow (Pimephales Promelas) to Examine Exposure Variables. Presented at Society of Environmental Toxicology & Chemistry, Portland, OR, November 14-18, 2004.
Abstract: Microarray technology has proven to be a useful tool for analyzing the transcriptome of various organisms representing conditions such as disease states, developmental stages, and responses to chemical exposure. Although most commercially available arrays are limited to organisms that have complete, or near complete, genome information, efforts directed at developing arrays for other model organisms have been gaining attention in an effort to fulfill experrimental and programmatic needs of academia, industry, and regulatory agencies. We describe some of the first experiments examining the use of custom made oligonucleotide arrays developed to assess transcriptome changes in a popular aquatic toxicity model, the fathead minnow (Pimephales promelas). Microarray results for adult tissues from several chemical exposures are presented. Use of guidelines following Microarray Gene Expression Data (MGED) Society standards such as MIAMI (minimal information about microarray experiments) and MAGE (microarry gene expression) for experimental design and validation are described with the perspective of producing data that are useful for environmental studies.

PRESENTATION Comparison of Qpcr Methods for the Detection of Vitellogenin Expression in Fathead Minnows 11/15/2004
Biales, A, C. Leutenegger, D. Riordan, V. deVlaming, J M. Lazorchak, AND D L. Lattier. Comparison of Qpcr Methods for the Detection of Vitellogenin Expression in Fathead Minnows. Presented at Society of Environmental Toxicology & Chemistry, Portland, OR, November 14-18, 2004.
Abstract: Male fathead minnows (FHM) normally express little if any of the egg yolk precursor protein vitellogenin (Vg). However, when exposed to estrogenic compounds such as 17a-ethynylestradiol (EE2), transcriptional levels of Vg rise dramatically and result in decreased fecundity and increased mortality rates. For this reason, measuring the expression of Vg in male FHM is a biologically relevant and useful tool for assessing exposure to estrogenic compounds. Recent advances in gene expressiion analysis have led to the establishment of two PCR-based methods, Sybrgreen and Taqman, which allow real-time monitoring of gene expression levels. These real-time platforms are both rapid and sensitive; however, their relative efficacy and applicability in the context of monitoring environmental quality has not been adequately evaluated. To this end, we compare the sensitivity, reproducibility and practical application of these two methods in gene expression monitoring. The Vg gene was cloned and used as a standard curve to determine the limit of sensitivity of Vg between the two experimental platforms. We also determined the reproducibility of quantification across a range of concentrations through comparison of standard deviations of replicate reactions. The ability to monitor Vg expression in multiple fish species with little manipulation of established protocols is desirable. Therefore, we determined the applicability of each QPCR protocol (Taqman or Sybrgreen) to monitor Vg expression in additional cyprinid fish species. Finally we have compared the limit of detection in male FHMs exposed to 2.5 ng/L, 5 ng/L or 10 ng/L EE2 using two different established protocols for both the Sybrgreen and Taqman methods to illustrate the importance of method optimization and to assess cross lab variability. Our results will have important implications in the establishment of gene expression based assays for monitoring water quality.

PRESENTATION Aromatase-B (Cyp 19b) Expression in Fathead Minnows (Pimephales Promelas) Exposed to Perfluorooctane (PFOS) and the Aromatase Inhibitor Fadrozole 11/15/2004
Knoebl, I, D Villeneuve, M. Kahl, K M. Jensen, D Kuehl, AND G Ankley. Aromatase-B (Cyp 19b) Expression in Fathead Minnows (Pimephales Promelas) Exposed to Perfluorooctane (PFOS) and the Aromatase Inhibitor Fadrozole. Presented at Society of Environmental Toxicology & Chemistry, Portland, OR, November 14-18, 2004.
Abstract: Perfluorooctane sulfonate (PFOS) is a fluorinated organic contaminant that is globally distributed in both humans and wildlife. PFOS belongs to a family of perfluorinated sulfonates that are highly persistent in the environment and have been commercially produced for over 40 years. These compounds are used in paint, polishes, food packaging, fire-fighting foams and fabric treatment. Although PFOS has been measured in a variety of wildlife, information as to its toxic mode of action (MOA) is scarce. PFOS may be an inhibitor of CYP19 aromatase, the P450 enzyme that converts testosterone to estradiol in vertebrates. To evaluate this possible MOA of PFOS in fish, we have developed a quantitative real-time PCR (TaqMan) assay to measure induction or inhibition of the aromatase gene in the brains and ovaries of fathead minnows in response to exposure to test chemicals. An aromatase cDNA sequence was isolated and cloned from the brains of fathead minnows and was used to construct a standard curve and to design sequence-specific primers and a probe. The assay was optimized and used to quantitate CYP19 gene expression in fish exposed to PFOS and to a known aromatase inhibitor, the pharmaceutical fadrozole. Fish were exposed to fadrozole (50, 17.7, 5.55, 1.85 ug/L) for 7 days, and to PFOS (.03, 0.3, 0.1, 1.0 mg/L) for 21 days. We present data comparing aromatase gene expression in brains and ovaries of the exposed fish, and make comparisons to aromatase enzyme activity in those tissues.

PRESENTATION Iterative Application of the EPA's Stressor Identification Guidance Discovers An Illicit Discharge and Residual Dam-Associated Impairment 11/15/2004
Cormier, S M., S B. Norton, G Suter, B Subramanian, C. Bellucci, AND G. Hoffman. Iterative Application of the EPA's Stressor Identification Guidance Discovers An Illicit Discharge and Residual Dam-Associated Impairment. Presented at Society of Environmental Toxicology & Chemistry, Portland, OR, November 14-18, 2004.
Abstract: EPA guidance for identifying causes of biological impairments is becoming more commonly applied by states to analyze evidence, logically determine causes, and document the process. One example is an investigation of biological impairment in the Willimantic River in Connecticut that occurred upstream from an NPDES regulated source. Initial analysis of six possible candidate causes indicated that none of the causes were sufficient to result in the level of effects at the site. Additional data, collected to identify the spatial extent of the impairment, revealed an episodic, illicit discharge of industrial wastewater. Biological condition improved following rerouting of the discharge to the waste treatment facility, providing a form of experimental evidence supporting a toxic cause of the biological impairment. Although improved, the biological diversity at the site remained at levels that precluded attainment of state biocriteria. The biological impairment was re-examined based on similarities of the site and four additional impaired sites and dissimilarities with four reference sites in the same waterhsed. All five impaired sites were located below old mill dams where water temperatures were elevated to levels reported to cause biological effects. The impoundments may also reduce summertime base flow and alter food resources towards more fine filterable particles. Hence, reservoirs may be reducing diversity at the impaired sites by more than one mechanism.

PRESENTATION Associations Between Measures of Bedded Sediments and Stream Life 11/15/2004
Cormier, S M., S. Niemela, AND J. Chirhart. Associations Between Measures of Bedded Sediments and Stream Life. Presented at Society of Environmental Toxicology & Chemistry, Portland, OR, November 14-18, 2004.
Abstract: Associations between causal agents and biological effects are necessary for the development of water quality criteria and, when combined with information from a site, can provide evidence for causal analysis. These associations can be obtained from controlled laboratory studies or from observational studies of natural systems. Quantile regression, regression using various binning procedures, and maximally observed biological performance conditional on stressor levels are among some of the approaches to quantify and illustrate effects observed in natural systems. These three methods were compared using data obtained by the Minnesota Pollution Control Agency in the St. Croix River Basin during 1996-2003. Fish and invertebrate metrics were analyzed along with measures of percent embeddedness, percent fines and depth of fines. Invertebrate metrics and percent fines were more sensitive measures of effects and exposure, than fish and percent embeddedness or depth of fines. This initial study did not evaluate organic matter, resolution of particle size below 1mm, stream gradient or other factors than may contribute to overall effects, and yet useful associations were identified that states and tribes might use to set criteria for regulation of excess sediment in streams and for TMDL cases where sediment is a candidate cause of biological impairment.

PRESENTATION Molecular, Protein and Histological Indicators in Fathead Minnows Exposedf to Ee2 in a Whole-Lake Dosing Experiment 11/15/2004
Lazorchak, J M., V. Palace, R. E. Evans, K. Wautier, R Flick, B E. Wiechman, K. Kidd, AND D L. Lattier. Molecular, Protein and Histological Indicators in Fathead Minnows Exposedf to Ee2 in a Whole-Lake Dosing Experiment. Presented at Society of Environmental Toxicology & Chemistry, Portland, OR, November 14-18, 2004.
Abstract: A whole-lake endocrine disruption experiment was conducted by Fisheries and Oceans Canada at the Experimental Lakes Area (ELA), northwestern Ontario from 2001 to 2003. This experiment examined population, organism, biochemical and cellular-level effects in lake trout, white sucker, fathead minnow and pearl dace exposed to environmentally-relevant concentrations of the synthetic estrogen, 17a-ethynylestradiol. The USEPA collaborated in this study by evaluating vitellogenin (Vtg) gene expression in male fathead minnow and male pearl dace livers from indigenous fish collected from the dosed and reference lakes. In addition, for the same collections of fathead minnows and pearl dace, concentrations of vitellogenin protein were determined in whole body homogenates using an indirect competitive ELISA assay and histological analyses were done on liver and gonads. Gene expression results indicate that male fathead minnows and pearl dace show induced vitellogenin gene expression throughout the dosing season and females of both fish show levels of gene expression beyond the normal breeding season. Vitellogenin protein levels were significantly elevated over background levels and over those in fish collected from control lakes. Histological sections of fathead minnow testes from Lake 260 revealed widespread fibrosis and inhibited development. In addition, EE2-exposed fish had kidney anomalies including edema within and between the kidney tubules, and hyaline deposits in the tubule cells. Liver tissues from fish from Lake 260 also had reduced glycogen stores and increased liver cell size when compared to baseline data from the same lake and reference lake fish.

PRESENTATION The Instability of Estrogenic Chemicals During Laboratory Static Exposure Conditions With Male Fathead Minnows 11/15/2004
Reddy, T V., J M. Lazorchak, M E. Smith, P E. Grimmett, D F. Bender, B E. Wiechman, AND D L. Lattier. The Instability of Estrogenic Chemicals During Laboratory Static Exposure Conditions With Male Fathead Minnows. Presented at Society of Environmental Toxicology & Chemistry, Portland, OR, November 14-18, 2004.
Abstract: Endocrine disrupting chemicals (EDCs) such as Para-nonylphenol (NP), estradiol (E2), estrone (E1), estriol (E3) and ethynylestradiol (EE2) are shown to be ubiquitous in surface waters, sediments and sludge. These EDCs are known to induce vitellogenin gene (Vg) expression in male fathead minnows. We have observed that the aforementioned EDCs are not stable during static laboratory conditions under high biomass to volume exposures at 25oC, with aeration at the rate of 100 bubbles minute-1 and a loading rate of 3 g fish Liter-1 instead of the standard rate of 0.5 g fish Liter-1. Male fat head minnows were exposed to NP at nominal concentrations for 24 hours and no detectable levels of vitellogenin gene (Vg) transcripts were seen. We checked the analytical values of NP in exposure water and found that measured values were only 2% of the nominal values. Additional experiments revealed that within 4 hours of exposure 98% of the NP was lost, of which 30% of nominal dose was bound to external body mucous of the fish and the remaining 68% was unaccounted. Subsequent analysis of E1, E2, E3 and EE2 under similar test conditions revealed that E1 and E3 showed losses similar to NP while, E2 and EE2 exhibited less concentration decline (20-30%). In order to maintain analytical concentrations close to the nominal concentrations, NP was supplemented continuously at appropriate concentrations by means of a peristaltic pump. Under this protocol, the final measured concentrations were close to 80% of the nominal dose and, in addition, Vg expression was observed in a dose dependant manner. These results show the importance of the exposure regimen in determining/interpreting the effects of EDCs in fish.

PRESENTATION The Use of Gene Arrays to Determine Temporal Gene Induction in Sheepshead Minnows Exposed to E2 11/15/2004
Knoebl, I, J. L. Blum, AND N Denslow. The Use of Gene Arrays to Determine Temporal Gene Induction in Sheepshead Minnows Exposed to E2. Presented at Society of Environmental Toxicology & Chemistry, Portland, OR, November 14-18, 2004.
Abstract: Gene arrays provide a means to study differential gene expression in fish exposed to environmental estrogens by providing a "snapshot" of the genes expressed at a given time. Such array data may also uncover previously unknown biochemical pathways affected by estrogenic compounds. To study the temporal induction of certain estrogen responsive genes, we have expanded an existing cDNA gene array {Larkin, 2002 #1342; Larkin, 2003 #1340} developed for the sheepshead minnow (SHM) (Cyprinodon variegates) to include over 200 additional genes that may be differentially regulated by EDCs. These genes were obtained from SHM liver cDNA libraries, suppressive subtractive hybridization libraries of liver mRNA from SHM exposed to known EDCs (methoxychlor, nonylphenol) or from differential display analyses. The gene arrays were used to examine temporal gene induction in livers of adult male SHM exposed to 100 ng/L and 500 ng/L 17 -estradiol for 6, 12, 24 or 48 hr. The estrogen receptor alpha gene was induced beginning at 6 hr, but the level of induction did not increase with longer exposure times. The estrogen responsive genes (vitellogenins [VTG 1, VTG2] and zona radiata [ZRP2, ZRP3]) were upregulated beginning 12 hr after exposure with the induction level increasing as exposure time increased. VTG1 was expressed at a 10-fold higher level than VTG2 after 48 hr. Other genes that appear to be upregulated over 48 hr include ATP synthase 6, serum amyloid protein, tryptophan 2,3 dioxygenase, a secretory phospholipase precursor and several unidentified genes. Several genes appear to be downregulated including fibrinogen, cytochrome b and cytochrome c oxidase. A number of genes increase after 6 and 12 hr of exposure, but are down-regulated to levels below those of unexposed fish after 48 hr. Those genes include chitinase, a scavenger receptor, C type lectin S and several unidentified genes. Quantitative real-time PCR (TaqMan) was used to validate the array results for VTG1, VTG2, ZRP2 and ZRP3. Through the use of gene arrays, temporal patterns of altered gene expression can be determined for key genes involved in metabolic pathways, and these changes can be quantified by real-time PCR.

PRESENTATION Xenobiotic Induced Organ-Specific Gene Expression and Macro/Microarray Development in Medaka (Oryzias Latipes) 11/09/2004
Bencic, D C., D. C. Volz, P. Chen, D. E. Hinton, AND S. W. Kullman. Xenobiotic Induced Organ-Specific Gene Expression and Macro/Microarray Development in Medaka (Oryzias Latipes). Presented at Society of Environmental Toxicology & Chemistry, Portland, OR, November 14 - 18, 2004.
Abstract: As part of an ongoing effort to understand and address the short and long-term consequences of increasing levels of environmental contaminants, we used suppressive subtractive hydridization (SSH) to develop gene expression profiles from Japanese medaka (Oryzias latipes) exposed to various prototypical xenobiotics. SSH analysis provided an unbiased, open-ended tool for screening gene expression change sin male medaka tissues following 48 h exposure to the endocrine disrupting compound 17B-estradiol (E2), the hypolipidemic pharmaceutical ciprofibrate (CF) or the persistent organic pollutant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The actions of these xenobiotics are typically mediated via very specific and different nuclear receptors: the estrogen receptor for E2, the peroxisome proliferator-activated receptor a for CF, and the aryl hydrocarbon receptor for TCDD. Furthermore, while each is known to strongly alter the expression of specific genes (E2, CF and TCDD strongly upregulate vitellogenin, peroxisome membrane protein 70, and cytochrome P450 1A, respectively), much less is known about organ-specific gene expression responses. We sequenced 1800 clones and subsequently identified approximately 600 differentially expressed, non-redundant genes from male medaka brain, liver, and testis following E2, CF or TCDD exposure. Of these total identified genes, less than 5% were shared among all three organs or xenobiotics, suggesting xenobiotic- and organ-specific responses at the level of gene expression. Many of the highly responsive genes and their corresponding proteins are being investigated as potentially novel markers of toxicity using approaches such as real-time RT-PCR, in situ hybridization, and immunohistochemistry. Furthermore, the results of these xenobiotic-iduced gene expression studies were used to develop a DNA macro-microarray to assess aquatic exposure and effect, with an emphasis on the linkage between gene expression and altered phenotype. The array technology will help us link molecular events and biochemical alterations following exposure and provide a mechanism to evaluate exposure and effects at both the individual and population levels.

PRESENTATION Discussions on Water Quality Assessment and Management: Ord Perspective 11/04/2004
Cormier, S M. Discussions on Water Quality Assessment and Management: Ord Perspective. Presented at Water Quality Assessment and Management Workshop, Columbus, OH, November 4, 2004.
Abstract: There is no abstract available for this product. If further information is requested, please refer to the bibliographic citation and contact the person listed under Contact field.

PRESENTATION Development of Standardized Large River Bioassessment Protocols (Lr-Bp) for Fish Assemblages 11/02/2004
Flotemersch, J E. Development of Standardized Large River Bioassessment Protocols (Lr-Bp) for Fish Assemblages. Presented at Southeastern Water Pollution Biologists Association, Opelika, AL, November 01 - 04, 2004.
Abstract: We conducted research comparing several methods currently in use for the bioassessment and monitoring of fish and benthic macroinvertebrate assemblages for large rivers. Fish data demonstrate that electrofishing 1000 m of shoreline is sufficient for bioassessments on boatable rivers similar to those in this study, regardless of whether the shoreline is along a single bank or distributed equally among paired banks. However, at sites with depths > 4 m, it may be advisable to employ nighttime electrofishing or increase day electrofishing distance to 2000 m. For benthic macroinvertebrates, we conducted a study using an experimental sampling method that is designed to overcome sampling limitations identified by previous research comparing several methods currently in use. Our objectives were to determine the appropriate number of sampling points needed, determine an appropriate laboratory subsample size, and to examine how varying reach lengths affect assemblage characteristics. Results indicated that, using the sampling method, a representative sample of the benthic macroinvertebrate fauna was collected by sampling both banks at 6 transects spaced at 100-m intervals over a 500-m distance. It is recommended that the field method be coupled with a fixed laboratory subsample size of 300 organisms for bioassessment purposes, with the recognition that a subsample size of 500 organisms may be needed to meet the objectives of some studies. It is likely this approach will over-sample sites of uniform composition, but the goal was to develop a standardized sampling protocol that would perform well across sites of differing habitat composition. However, resulting data should be interpreted with consideration of appropriate classification systems (e.g., ecoregion, free-flowing vs. impounded, tidal vs. non-tidal).

PRESENTATION Development of Large River Bioassessment Protocols (Lr-Bps) for Benthic Macroinvertebrates in EPA Region 5 11/02/2004
Flotemersch, J E. Development of Large River Bioassessment Protocols (Lr-Bps) for Benthic Macroinvertebrates in EPA Region 5. Presented at Southeastern Water Pollution Biologists Association, Opelika, AL, November 01 - 04, 2004.
Abstract: Non-wadeable rivers have been largely overlooked by bioassessment programs because of sampling difficulties and a lack of appropriate methods and biological indicators. We are in the process of developing a Large River Bioassessment Protocol (LR-BP) for sampling macroinvertebrate assemblages. The LR-BP is a semi-quantitative, multi-habitat method where samples are collected from 10-m sample zones on each bank at 6 transects spaced 100-m apart over a distance of 500-m. In summer 2004 and 2005, the LR-BP will be used to sample 25 probabilistic sites on each of six rivers of differing landuse type in USEPA Region 5. Our objectives are to refine the LR-BP and to compare it with other large river macroinvertebrate sampling methods at a subset of sites on each river. We are also collecting snag samples on each river in an attempt to develop a rapid bioreconaissance tool for monitoring agencies. Expected outcomes of this research include probability-based assessments of the target rivers, standardized large river macroinvertebrate sampling methods, indicator development, and a comparison of existing sampling methods. This work will also be used to produce a summary of large river sampling methods that documents associated performance characteristics for each. This research was funded by a Regional Methods Initiative (RMI) grant to NERL/EERD from EPA Region 5.

PRESENTATION Development of Standardized Large River Bioassessment Protocols (Lr-Bp) for Benthic Macroinvertebrate Assemblages 11/02/2004
Flotemersch, J E. Development of Standardized Large River Bioassessment Protocols (Lr-Bp) for Benthic Macroinvertebrate Assemblages. Presented at Southeastern Water Pollution Biologists Association, Opelika, AL, November 01 - 04, 2004.
Abstract: We conducted research comparing several methods currently in use for the bioassessment and monitoring of fish and benthic macroinvertebrate assemblages of large rivers. Fish data demonstrate that electrofishing 1000 m of shoreline is sufficient for bioassessments on boatable rivers similar to those in this study, regardless of whether the shoreline is along a single bank or distributed equally among paired banks. However, at sites with depths > 4 m, it may be advisable to employ nighttime electrofishing or increase day electrofishing distance to 2000 m. For benthic macroinvertebrates, we conducted a study using an experimental sampling method that is designed to overcome sampling limitations identified by previous research comparing several methods currently in use. Our objectives were to determine the appropriate number of sampling points needed, determine an appropriate laboratory subsample size, and to examine how varying reach lengths affect assemblage characteristics. Results indicated that, using the sampling method, a representative sample of the benthic macroinvertebrate fauna was collected by sampling both banks at transects spaced at 100-m intervals over a 500-m distance. It is recommended that the field method be coupled with a fixed laboratory subsample size of 300 organisms for bioassessment purposes, with the recognition that a subsample size of 500 organisms may be needed to meet the objectives of some studies. It is likely this approach will over-sample sites of uniform composition, but the goal was to develop a standardized sampling protocol that would perform well across sites of differing habitat composition. However, resulting data should be interpreted with consideration of appropriate classification systems (e.g., ecoregion, free-flowing vs. impounded, tidal vs. non-tidal).

PRESENTATION Large and Great Rivers: New Assessment Tools 11/02/2004
Flotemersch, J E. Large and Great Rivers: New Assessment Tools. Presented at Southeastern Water Pollution Biologists Association, Opelika, AL, November 01 - 04, 2004.
Abstract: The Ecological Exposure Research Division has been conducting research to support the development of the next generation of bioassessment and monitoring tools for large and great rivers. Focus has largely been on the development of standardized protocols for the traditional indicators: algae, benthic macroinvertebrates, fish assemblages, and physical habitat. Several projects are currently underway using and refining the protocols across the nation. Research is also in progress for several novel indicators. Tissue samples are being collected in numerous studies to develop indicators for endocrine disruptors. Multiple projects have also been conducted using remotely sensed data collected via airplane and satellite-based spectrographic imagers. Efforts have been focused on correlating water quality spectral signatures for chlorophyll-a, turbidity, total suspended solids and phosphorus with land use and ecological condition of large rivers. Goals of this work include development of spectral tools for a suite of water quality parameters that can be used to assess entire rivers in near real time for pennies on the dollar and more frequently and at less cost then currently possible.

PRESENTATION Ecological Epidemiology: A Means to Safeguard Services of Nature That Sustain Human Welfare 10/13/2004
Cormier, S M., G Suter, AND S B. Norton. Ecological Epidemiology: A Means to Safeguard Services of Nature That Sustain Human Welfare. Presented at NATO Workshop of Ecotoxicology, Ecological Risk Assessment and Multiple Stressors, Poros, Greece, October 12-15, 2004.
Abstract: The services provided by nature are required to sustain human life and enhance its quality. Hence, environmental security must come from protecting those services. Ecological risk assessment can predict and estimate effects of proposed actions, but it is insufficient alone for two reasons. First, it can fail because of inadequate application, unforeseen stressors, or unpredictable effects. Second, in many cases the ability of nature to provide services that sustain life is already impaired, resulting in reduced human welfare. For these reasons, environmental security requires the development of ecological epidemiology, a science that will identify impaired services of nature and determine the causes of impairment so that remediation and restoration can occur. A method for causal analysis, developed to identify causes of impairment in aquatic ecosystems, may provide a template that can be adapted to identify the causes of diminished services of nature and the resulting reductions in human welfare. Some of the challenges for adapting the existing method include explicitly defining natural services required to sustain human life, appropriately matching the scale of the analysis to the ecological processes that deliver those services, and possibly customizing the logical considerations used in causal analysis. Advancing the science of ecological epidemiology holds the promise of helping scientists frame and guide rational debate, providing a sound basis from which to launch risk assessment and risk management scenarios, and ultimately informing environmental decision-making that affects human welfare, development and environmental security within acceptable risks.

PRESENTATION Studies in Southeastern Aquatic Ecosystems 09/29/2004
WALTERS, DAVID M. Studies in Southeastern Aquatic Ecosystems. Presented at 2004 Region 4 Biology Meeting, Atlanta, GA, September 28 - 29, 2004.
Abstract: This presentation provides an overview of selected ORD studies within Region 4. We are intiating a study of red shiners, an invasive minnow species of streams in the Atlanta metropolitan area. This noxious species has negatively impacted native fishes in the region, and our research is designed to understand the genetic, behavioral, and environmental mechanisms contributing to its spread. A second project, which is ongoing, addresses polychlorinated biphenyl (PCB) contamination in a stream near Clemson, SC. Our goals for this project are to determine the extent and magnitude of contamination, to identify pathways of PCBs through stream food webs, and to develop tools for monitoring the success of mitigation efforts.

PRESENTATION Landscape Influences on Non-Point Sourced Nutrients for Agricultural Watersheds in Ohio 09/27/2004
Daniel, F B. Landscape Influences on Non-Point Sourced Nutrients for Agricultural Watersheds in Ohio. Presented at 12th National Nonpoint Source Monitoring Workshop, Ocean City, MD, September 26-30, 2004.
Abstract: In stream nutrient concentrations for a set of 35 small agricultural subwatersheds in southwestern Ohio have been monitored with respect to both water quality and biological integrity measures over a five year period. The dominant land cover in all of these subwatersheds is row crop agriculture. However, approximately one half (19) of the subwatersheds are located in relatively low clay-contented soils derived from glacial tills whereas the remainder (16) are located in a landscape dominated by older soils with a significantly higher clay content, greater erodibility, and lower permeability. The observed in-stream nutrient concentrations in these streams can be explained in terms of a set of landscape measures, of both anthropogenic and geophysical, in the surrounding catchments. These studies indicate the various classes of landscape measures interact to control the export of nutrients from agricultural land.

PRESENTATION Ecotoxicogenomics: the Use of 'omics Technologies in Aquatic Toxicology 09/22/2004
Miracle, A L. Ecotoxicogenomics: the Use of 'omics Technologies in Aquatic Toxicology. Presented at Eugene Applebaum College of Pharmacy and Health Sciences, Detroit, MI, September 22, 2004.
Abstract: There is no abstract available for this product. If further information is requested, please refer to the bibliographic citation and contact the person listed under Contact field.

PRESENTATION Changes in Genetic Diversity of a White Sucker Population Following Experimental Whole-Lake Acidification 08/22/2004
Bagley, M J., S A. Christ, K. H. Mills, AND S. M. Chalanchuk. Changes in Genetic Diversity of a White Sucker Population Following Experimental Whole-Lake Acidification. Presented at American Fisheries Society, Madison, WI, August 21-26, 2004.
Abstract: Despite great strides to reduce sulfur dioxide emissions over the last decade, acid precipitation remains a persistent threat to North American fish communities. A demographic analysis of white suckers in an experimentally acidified oligotrophic lake in northwest Ontario demonstrated complete recruitment failure in three years when the pH was between 5.0 and 5.2 and highly variable recruitment during subsequent recovery. We exploited archived fin rays that had been used for age analysis to evaluate genetic changes during the acidification (1976-1983) and recovery (1984-1996) phases of the experiment. We hypothesized that serial recruitment failures and high recruitment variability would create a population bottleneck, reducing genetic diversity relative to white suckers in a nearby reference lake. Allelic distributions at twelve microsatellite loci were evaluated for 1150 samples collected from the acidified lake and 745 samples from the control lake, representing year classes between 1957 and 1994. Surprisingly, genetic diversity greatly increased in the first year of acidificatiion and remained relatively stable thereafter. One locus which was intially monomorphic segregated for ten alleles following acidification. The observed changes in genetic diversity suggest that the most important effect of acidification may have been to reduce barriers to effective immigration into the lake.

PRESENTATION Molecular Genetic Approaches to Pest and Nontarget Population Monitoring 08/04/2004
Stolz, U. Molecular Genetic Approaches to Pest and Nontarget Population Monitoring. Presented at Conference on Development of Strategic Monitoring Programs for Ecological Impact from Plant-Incorporated Protectants, Arlington, VA, August 3-5, 2004.
Abstract: The U.S. Environmental Protection Agency has interest in a number of applications of genetic monitoring methodologies. Genetic monitoring in agroecosystems can provide valuable environmental information regarding both traditional and novel pesticides. One group of pesticides of particular interest is plant incorporated protectants (PIPs), which are genetically modified (GM) plants that express insecticidal proteins (e.g., Bt corn and Bt cotton). It has been argued that PIP crops have the potential to cause population or ecosystem-level changes through a number of mechanisms (Ervin et al. 2003). These changes include rapid resistance evolution in pest populations, ecological harm caused by transgenes from GM crops escaping into close relatives via crop-relative interbreeding, and negative effects on non-target organisms. Regulatory agencies such as the EPA and USDA have evaluated the environmental risks of PIP crops and judged them acceptable. However, additional data would help to support both regulatory confidence as well as public confidence in the long term environmental safety of PIPs. Because empirical data to support assessments of long term ecological risks of PIPs is an immediate need of the EPA, I believe applications of genetic monitoring for examining the long term ecological effects of PIPs should be developed as soon as possible.

PRESENTATION Preliminary Analysis of Common Loon Genetic Structure in North America Based on Five Microsatellite Loci 07/31/2004
McMillan, A., M J. Bagley, AND D. Evers. Preliminary Analysis of Common Loon Genetic Structure in North America Based on Five Microsatellite Loci. Presented at Society for Conservation Biology, New York, NY, July 30-August 2, 2004.
Abstract: This study seeks to determine fine-scale genetic structure of Common Loon breeding populations in order to link wintering birds with their breeding regions. Common Loons are large piscivorous birds that breed in lakes of northern North America and Iceland. Loons are highly philopatric and territorial in breeding areas and are susceptible to mercury poisoning, lake acidification and other threats across much of this region. Wintering loon populations originate from a mix of breeding regions. In North America, wintering populations are found primarily in nearshore coastal environments and these birds are susceptible to oil spills. Loons also are threatened by the current botulism poisoning outbreak, which has killed thousands of loons in the Great Lakes. Despite significant demographic data, little is known about the population genetic structure of Common Loons. Preliminary analysis using five polymorphic microsatellite loci demonstrated strong differentiation between loons in eastern and western North America. Differentiation among five putative eastern loon breeding populations was also identified. Differences were found in four of ten pairwise comparisons. The information developed on loon population structure will be crucial for understanding year-round impacts on these birds.

PRESENTATION Development of Aquatic Models for Testing the Relationship Between Genetic Diversity and Population Extinction Risk 06/27/2004
Markert, J, M J. Bagley, AND D Nacci. Development of Aquatic Models for Testing the Relationship Between Genetic Diversity and Population Extinction Risk. Presented at Evolution 2004 Conference, Fort Collins, CO, June 26-30, 2004.
Abstract: The relationship between population adaptive potential and extinction risk in a changing environment is not well understood. Although the expectation is that genetic diversity is directly related to the capacity of populations to adapt, the statistical and predictive aspects of this relationship in real populations are not well known. From a conservation perspective, it is useful to understand this relationship so that genetic data may be incorporated into population viability models which traditionally rely on demographic and ecological data. Here we present an experimental design using the freshwater amphipod species Hyalella azteca and the marine mysid Americamysis bahia. These organisms differ with respect to demographic and biogeographic constraints, and present a useful contrast with Drosophila and Tribolium laboratory models. The design involves creating inbred lines from diverse native populations. Individuals from each inbred line will be combined to form "synthetic" populations from either three or six randomly selected inbred lines in order to generate experimental populations with controlled levels of genetic diversity, and the entire set will be replicated ten times. The adaptive capacity of these "synthetic" and inbred populations will be compared to that of native populations and an "admixed" population containing genes from all sampled populations. Differences in population extinction times will be interpreted as differences in population adaptive capacity, and provide data for formal population viability analyses (PVA's). By experimentally manipulating levels of population genetic diversity, we will be able to formally assess both the relationship between genetic diversity and population viability in an increasingly hostile environment and also determine the statistical efficiency of various classes of genetic markers.

PRESENTATION Abscissa Assessment With Algae: A Comparison of Local and Landscape Impairment Measures for Biological Assessment Using Benthic Diatoms 06/07/2004
Lane, C R. Abscissa Assessment With Algae: A Comparison of Local and Landscape Impairment Measures for Biological Assessment Using Benthic Diatoms. Presented at North American Benthological Society, Vancouver, Canada, June 6-10, 2004.
Abstract: The development of rigorous biological assessments is dependent upon well-constructed abscissa, and various methods, both subjective and objective, exist to measure expected impairment at both the landscape and local scale. A new, landscape-scale method has recently been offered that allows for the combination of disparate factors into a single value (e.g., impervious surfaces, low-density residential, improved pasture, natural area): the Landscape Development Intensity index, or LDI. The LDI is calculated using GIS and published development coefficients, which are based on "emergy" calculations.
In this study, benthic diatoms from 70 isolated depressional herbaceous wetlands located throughout peninsular Florida were sampled along a gradient of human disturbance. Analyses were conducted at both the local and landscape scale to determine the efficacy of the LDI vis-a-vis locally measured physical and chemical variables of the water column and sediment (e.g., soil TP, water pH, specific conductivity). Multivariate analyses suggested that the LDI is reflective of benthic diatom community composition within isolated depressional wetlands, although individual variables measured on-site, such as specific conductivity or water total phosphorus, may be more strongly correlated.

PRESENTATION Natural and Human Factors Structuring Fish Assemblages in West Virginia Wadeable Streams 06/07/2004
WALTERS, DAVID M. AND F H. McCormick. Natural and Human Factors Structuring Fish Assemblages in West Virginia Wadeable Streams. Presented at North American Benthological Society, Vancouver, BC, CANADA, June 06 - 10, 2004.
Abstract: We surveyed fishes and environmental variables in 119 stream basins to identify natural and anthropogenic factors structuring fish assemblages. We collected fishes and physico-chemical variables using standardized EPA methods and compiled basin characteristics (e.g., land cover) from digital coverages. Nonmetric multidimensional scaling, NMS, analysis identified two axes that explained 70% of among-site variation in fishes. Axis 1 (31%) positively correlated with pH and basin area and corresponded with a shift from tolerant to sensitive species as pH increased. Axis 2 (39%) strongly correlated with temperature, elevation, and slope. High elevation, cold, steep streams were dominated by salmonids, cottids, and Rhinichthys spp. that gradually changed to diverse assemblages of cyprinids, percids, catostomids, ictalurids, and centrarchids at lower elevations. Land cover variables were weakly correlated with NMS axes but were significantly correlated with changes in water quality (e.g., conductivity, SO4, and NO2/NO3 increased with mining and urbanization). Our findings suggest that land use effects may be difficult to detect in systems characterized by strong abiotic controls on assemblage structure and that changes in stream chemistry precede gross changes in fish assemblages.

PRESENTATION Temporal Variation in Ohio River Macroinvertebrates: A Historical Rock Basket Comparison, 1960's to Present 06/07/2004
Wooten, M. S., E. B. Emery, AND B R. Johnson. Temporal Variation in Ohio River Macroinvertebrates: A Historical Rock Basket Comparison, 1960's to Present. Presented at North American Benthological Society, Vancouver, Canada, June 6-10, 2004.
Abstract: Collection of representative macroinvertebrate samples has historically been a problem for researchers working on the Ohio River. The USEPA utilized rock basket artificial substrates to sample benthic assemblages from 1964-1971. By this method, a steel basket (7" diameter, 11" long) was filled with 2 - 3 inch limestone aggregate and suspended by steel cable in 2 - 3 feet of water. The baskets were generally suspended from lock chamber walls, although other permanent structures, such as concrete water intakes, were also used. The objective of this study was to repeat these rock basket surveys in order to accurately observe temporal trends within the macroinvertebrate community. Rock baskets were deployed in late summer 2002 in the same general areas of the historical studies and were allowed to colonize for the same six week time period. Macroinvertebrates collected from baskets were examined using a community similarity index, nonmetric multidimensional scaling, and a draft macroinvertebrate index developed for the Ohio River. Results indicated the 2002 data were distinct from historic data and showed increases in taxa richness, sensitive species, and index scores. Our findings indicate marked improvement in the macroinvertebrate community since passage of the 1972 Clean Water Act.

PRESENTATION The Effects of Riparian Management on Detritus Processing and Invertebrate Assemblages in Coastal Plain Intermittent Streams 06/07/2004
Fritz, K M. AND J. W. Feminella. The Effects of Riparian Management on Detritus Processing and Invertebrate Assemblages in Coastal Plain Intermittent Streams. Presented at North American Benthological Society, Vancouver, Canada, June 6-10, 2004.
Abstract: Silviculture is the primary land use within many Coastal Plain watersheds of the southeastern United States, where most forested wetlands are found along headwater intermittent streams. Our study compared invertebrate assemblages and breakdown of buried detritus (leaves, wood, and roots) among three riparian management treatments (clear cut, thinned, and intact). Decay rates did not differ among riparian treatments or detritus types over the one year incubation. Slow decay rates were associated initially with anaerobic conditions within sandy sediments and later with dry conditions. Abundance, diversity and biomass of invertebrates varied among riparian treatments and substrate types, but not consistently so across sampling periods. When differences occurred, ivnertebrate variables from the clear cut treatment were significantly lower than those from intact and thinned treatments, possibly from extremely high and variable temperature regimes in this treatment. Assemblages from the clear cut treatment became more dissimilar from other treatments over time. Patterns of assemblage structure were driven by riparian treatment and seasonal succession rather than substrate type. Riparian clear cutting appears to affect invertebrate assemblages inhabiting both aquatic and terrestrial phases of the intermittence cycle.

PRESENTATION Toxicogenomics as a Tool to Assess Exposure of Fish to Environmental Pollutants 06/02/2004
Knoebl, I. Toxicogenomics as a Tool to Assess Exposure of Fish to Environmental Pollutants. Presented at Science Forum 2004, Washington, DC, June 1-3, 2004.
Abstract: Molecular biological techniques such as gene arrays and quantitative real-time PCR are becoming important tools to study alterations in normal gene expression in fish and other wildlife exposed to such pollutants as endocrine disrupting chemicals (EDCs). An important function for these tools is the ability to translate from the laboratory to field conditions to assess exposure to EDCs and other pollutants in wild fish. Detection of anomalies at the genomic level will enable screening methods to identify toxic effects soon after exposure, at the molecular level, before they are manifested at the tissues, organs, individuals or population level. One area of concern in which in which these methods may be useful is in the assessment of concentrated animal feeding operation (CAFO) effluents on gene expression in fish. The results of this work will not only provide information as to the effects of CAFO effluents on gene expression in fish, but will also be an initial field validation of the fathead minnow gene array currently in development at EERD. It is hoped that the gene array can and will be used by EPA Regions, states, tribes and other monitoring agencies in field applications to screen for EDCs and other pollutants that alter normal gene expression in fish.

PRESENTATION Assessing Headwater Streams: Linking Landscapes to Stream Networks 06/02/2004
Fritz, K M., B R. Johnson, D M. Walters, AND J E. Flotemersch. Assessing Headwater Streams: Linking Landscapes to Stream Networks. Presented at Science Forum 2004, Washington, DC, June 1-3, 2004.
Abstract: Headwater streams represent a significant land-water boundary and drain 70-80% of the landscape. Headwater streams are vital components to drainage systems and are directly linked to our downstream rivers and lakes. However, alteration and loss of headwater streams have occurred without an understanding of the consequences to larger downstream waterbodies. Furthermore, headwater streams provide a more direct measure of land use effects because they drain smaller areas with less land use complexity than their larger counterparts, and therefore headwater may also be useful for identifying specific causes of water quality impairment. Drying is common in these systems and this may confound our ability to use traditional assessment methods. Understanding associations between the distribution of aquatic organisms and flow permanence is the first step in developing biocriteria for our nation's most abundant running waters. The Ecological Exposure Research Division is developing field protocols and identifying physical and biological indicators of flow permanence for headwater streams. The protocols we are developing will allow states, regions, and tribes to better classify and assess water quality in headwater streams. Through this research project the USEPA will have the tools to begin assessing condition appropriately for the majority of streams and stream miles in the United States. Protecting and improving the condition of headwater streams will result in better water quality in downstream water bodies.

PRESENTATION Red Shiner Invasion of Southeastern Streams: Dynamics and Ecological Consequences 06/02/2004
WALTERS, DAVID M., M Blum, AND B Rashleigh. Red Shiner Invasion of Southeastern Streams: Dynamics and Ecological Consequences. Presented at Science Forum 2004, Washington, DC, June 01 - 03, 2004.
Abstract: Red shiners, a minnow species native to streams of the central U.S., are spreading to other regions due to their widespread use as a bait-fish. Their expansion into new habitats comes at the expense of their native relatives. Red shiners are aggressive competitors for food and spawning habitat and interbreed with other species, swamping native populations with their own genes. These combined impacts have resulted in local extinction of native species. Red shiners were recently introduced into several Southeastern rivers and now pose a serious threat to the region's unique and diverse community of native fishes. We have initiated a study to determine how human disturbance and other environmental factors affect the spread of red shiners. This research will also identify the behavioral and genetic components of native species displacement. Our research supports the Agency's mission to protect biological integrity by providing the tools to forecast the spread of this invasive species, by identifying native, at-risk species, and by offering management solutions to curb expansion of red shiners into uninvaded watersheds.

PRESENTATION Targeted Screening for Invasive Species in Ballast: Genomic Approaches 06/02/2004
Blum, M. Targeted Screening for Invasive Species in Ballast: Genomic Approaches. Presented at Science Forum 2004, Washington, DC, June 1-3, 2004.
Abstract: There is no abstract available for this product. If further information is requested, please refer to the bibliographic citation and contact the person listed under Contact field.

PRESENTATION Molecular Indicators of Genetic Diversity: Utility at Local and Regional Scales 05/19/2004
Bagley, M J. Molecular Indicators of Genetic Diversity: Utility at Local and Regional Scales. Presented at Workshop on Environmental Indicators, Kansas City, MO, May 19, 2004.
Abstract: There is no abstract available for this product. If further information is requested, please refer to the bibliographic citation and contact the person listed under Contact field.

PRESENTATION A Comparison of Single and Multiple Habitat Rapid Bioassessment Sampling Methods for Macroinvertebrates in Piedmont and Northern Piedmont Streams 05/17/2004
Blocksom, K A., J E. Flotemersch, AND M. Passmore. A Comparison of Single and Multiple Habitat Rapid Bioassessment Sampling Methods for Macroinvertebrates in Piedmont and Northern Piedmont Streams. Presented at 4th National Monitoring Conference, Chattanooga, TN, May 17-20, 2004.
Abstract: Stream macroinvertebrate collection methods described in the Rapid Bioassessment Protocols (RBPs) have been used widely throughout the U.S. The first edition of the RBP manual in 1989 described a single habitat approach that focused on riffles and runs, where macroinvertebrate diversity and abundance is high. This approach was adopted by many states, tribes and regions. Many scientists interpreted the revised RBP protocol published in 1999 as a recommendation for multiple habitat sampling. However, no direct comparison of the two RBP protocols was presented in the second edition, and there were no recommendations for reconciling baseline data collected using the single habitat method with data collected using the multiple habitat method. As a result, scientists have been reluctant to switch from the single habitat approach, regardless of the merits that may exist in adopting the multiple habitat approach. In this study, both the single and multiple habitat methods were performed at each of 41 sites in the Piedmont and Northern Piedmont ecoregions. Differences between methods in collected macroinvertebrate assemblages will be examined using a newly developed multimetric index for Virginia and its component metrics, as well as taxonomic similarity indices. The influence of abiotic factors and site condition on these differences will be examined to determine conditions under which the two methods collect similar samples.

PRESENTATION Aquatic Impacts Study of Mountaintop Mining and Valley Fill Operations in West Virginia 05/14/2004
Fulk, F A. Aquatic Impacts Study of Mountaintop Mining and Valley Fill Operations in West Virginia. Presented at Ohio Valley Regional Chapter of SETAC, Columbus, OH, May 13-14, 2004.
Abstract: The practice of mountaintop mining and valley fill operations in West Virginia is fraught with controversy. In 1999, EPA, along with several state and federal agencies, initiated an environmental impact study (EIS) to investigate the economic, social and ecological impacts of the mountaintop mining and valley fill operations in the southwest corner of West Virginia. This presentation will focus on the aquatic impacts study that was included in the overall EIS. There were two primary objectives of the aquatic impacts study: (1) is the biological condition of streams in areas with MTM/VF operations degraded compared to the condition of streams in unmined areas; and (2) are there "additive" biological impacts in streams where multiple fills are located?
To address the primary objectives of the study, fish, macroinvertebrates, water samples and habitat information were collected from sites in five watersheds within the MTM/VF region of West Virginia. The water chemistry and habitat data were also used to address a third objective: do chemical and physical habitat factors contribute to any potential differences in stream condition detected for fish and invertebrates? Insights gained from the third objective may provide information to develop guidance to "minimize, to the maximum extent practicable, the adverse environmental effects to the waters of the United States and to fish and wildlife resources from mountaintop mining operations, and to environmental resources that could be affected by the size and location of fill material in valley fill sites."

PRESENTATION Using Landscape Indicators to Predict Pesticide and Nutrient Concentrations and Aquatic Biological Conditions in Small Streams in the Midwest 05/11/2004
Stark, J. R., D. L. Lorenz, S. P. Wente, T. A. Winterstein, A M. Pitchford, AND M D. Moeykens. Using Landscape Indicators to Predict Pesticide and Nutrient Concentrations and Aquatic Biological Conditions in Small Streams in the Midwest. Presented at Interagency Nutrient Issues and Science Strategy Meeting, Bloomington, MN, May 10-11, 2004.
Abstract: There is no abstract available for this product. If further information is requested, please refer to the bibliographic citation and contact the person listed under Contact field.

PRESENTATION 2003 Gene Expression Findings, Canadian Whole Lake Dosing Study 04/13/2004
Lazorchak, J M., R Flick, D L. Lattier, B E. Wiechman, M E. Smith, AND D A. Gordon. 2003 Gene Expression Findings, Canadian Whole Lake Dosing Study. Presented at Whole Lake Dosing Study Collaborators' Meeting, Winnepeg, Canada, April 13-14, 2004.
Abstract: There is no abstract available for this product. If further information is requested, please refer to the bibliographic citation and contact the person listed under Contact field.

PRESENTATION A Comparison of Multiple and Single Habitat Rapid Bioassessment Protocols (Rbps) for Collecting Benthic Macroinvertebrates in Wadeable Streams 03/25/2004
Autrey, B C., J E. Flotemersch, AND K A. Blocksom. A Comparison of Multiple and Single Habitat Rapid Bioassessment Protocols (Rbps) for Collecting Benthic Macroinvertebrates in Wadeable Streams. Presented at Mid-Atlantic Water Pollution Biology Workshop, Berkeley Springs, WV, March 24-26, 2004.
Abstract: There is no abstract available for this product. If further information is requested, please refer to the bibliographic citation and contact the person listed under Contact field.

PRESENTATION Multivariate Statistics Workshop 03/25/2004
Blocksom, K A. AND M. J. Paul. Multivariate Statistics Workshop. Presented at Mid-Atlantic Water Pollution Biology Workshop, Berkeley Springs, WV, March 24-26, 2004.
Abstract: There is no abstract available for this product. If further information is requested, please refer to the bibliographic citation and contact the person listed under Contact field.

PRESENTATION Development of a Fish Index of Biotic Integrity for Assessing Wadeable Streams in West Virginia 03/25/2004
WALTERS, DAVID M., F. H. MCCORMICK, AND D. CINCOTTA. Development of a Fish Index of Biotic Integrity for Assessing Wadeable Streams in West Virginia. Presented at Mid-Atlantic Water Pollution Biology Workshop, Berkeley Springs, WV, March 24-26, 2004.
Abstract: We used Environmental Monitoring Assessment Program (EMAP) data from the Mid-Atlantic Highlands Assessment (MAHA) to develop an Index of Biotic Integrity (IBI) for West Virginia (WV). Standard techniques were used to evaluate a candidate pool of 58 fish assemblage metrics. Responses of metrics to anthropogenic disturbance were analyzed at the regional and the state scale. Our results show that the existing IBI for the Mid-Atlantic region is applicable to WV for stream assessment. As part of a Regional Applied Research Effort (RARE) and Regional-EMAP project, we surveyed an additional 119 streams in WV to test the IBI and determine if it is applicable across a range of stream temperatures. Stream temperature data collected at the time of fish collection was unrelated to IBI scores. In addition, hourly temperature data from 84 sites were used to classify streams as cold and warm water streams. We found no differences in IBI scores between cool and warm streams, but sample size was too low to draw conclusions for cold streams. Variability in IBI scores for warm streams was low and cumulative distribution functions of the IBIs showed that RARE sites scored higher than those from previous EMAP studies. Higher scores for rare sites may be related to differences in the site selection process, differences in fish sampling protocols between EMAP and the WV Division of Natural Resources, or differences in disturbance gradients among EMAP and RARE sites.

PRESENTATION Headwater Intermittent Stream Workshop (Mid-Atlantic Wpbw) 03/24/2004
FRITZ, K. M. AND DAVID M. WALTERS. Headwater Intermittent Stream Workshop (Mid-Atlantic Wpbw). Presented at Mid-Atlantic Pollution Biology Workshop, Berkeley Springs, WV, March 24 - 26, 2004.
Abstract: There is no abstract available for this product. If further information is requested, please refer to the bibliographic citation and contact the person listed under Contact field.

PRESENTATION Biological Indicators in Stream Monitoring: What Are the Fish, Bugs and Algae Telling Us? 03/23/2004
Hill, B H., F H. McCormick, AND K A. Blocksom. Biological Indicators in Stream Monitoring: What Are the Fish, Bugs and Algae Telling Us? Presented at Minnesota Water 2004 Conference, Minneapolis, MN, March 23-24, 2004.
Abstract: There is no abstract available for this product. If further information is requested, please refer to the bibliographic citation and contact the person listed under Contact field.

PRESENTATION An EPA Sponsored Literature Review Database to Support Stressor Identification 03/18/2004
Jessup, B., C. Cresswell, P ShawAllen, B Subramanian, AND S M. Cormier. An EPA Sponsored Literature Review Database to Support Stressor Identification. Presented at New England Association of Environmental Biologists, Hancock, MA, March 17-19, 2004.
Abstract: The Causal Analysis/Diagnosis Decision Information System (CADDIS) is an EPA decision-support system currently under development for evaluating the biological impact of stressors on water bodies. In support of CADDIS, EPA is developing CADLIT, a searchable database of the scientific literature that will allow interactive searching of published relationships between sources, stressors, and responses. CADLIT is currently being populated with reviews of literature that include metals, clean sediment, and inorganic contaminants as stressors. When it is complete, investigators will be able to access and query the database through a CADDIS website. Evidence of consistent associations of exposures and responses, analogous experiments, stressor/response thresholds, and plausible mechanisms of stressor effects will be accessible for the investigator's source, stressor, response, and location of interest. Retrievable data and meta-data include study location, pollutant source, study design, sample collection methods, exposure characteristics, response type, response measurement, and response organism or taxa group. Reviewers summarize the contribution of each paper to stressor identification in a short paragraph and listing of applicable lines of evidence. This presentation will demonstrate the data input and querying capabilities of the CADLIT prototype.

PRESENTATION Research Supporting the Bioassessment of Large Rivers 03/18/2004
Flotemersch, J E. Research Supporting the Bioassessment of Large Rivers. Presented at New England Association of Environmental Biologists, Hancock, MA, March 17 - 19, 2004.
Abstract: There is no abstract available for this product. If further information is requested, please refer to the bibliographic citation and contact the person listed under Contact field.

PRESENTATION Causal Analysis and Adaptive Management in the Willimantic River, Connecticut That Led to Improved Benthic Community Condition 03/18/2004
Cormier, S M., C. Bellucci, G. Hoffman, G W. Suter, S B. Norton, B Subramanian, AND B C. Autrey. Causal Analysis and Adaptive Management in the Willimantic River, Connecticut That Led to Improved Benthic Community Condition. Presented at New England Association of Environmental Biologists, Hancock, MA, March 17-19, 2004.
Abstract: Determining the cause of a biological impairment can be a daunting task, one that is complicated by multiple possible causes, spotty or irregularly collected data, natural variations due to season, weather, land formation and other factors, and a lack of basic scientific information needed to diagnose an environmental problem. Nevertheless, probable causes of biological impairments can be determined and findings can be used to reduce or remove detrimental causes and improve condition of biological communities. For example, a 2.4 km segment of the Willimantic River in northeastern Connecticut was listed as impaired in 1998 based on a review of aquatic toxicity monitoring reports submitted to the Connecticut Department of Environmental Protection from the Stafford publicly owned treatment works (POTW). Calculations showed a high probability of exceeding Connecticut's water quality criteria for copper, lead and zinc. Subsequent monitoring in the fall of 1999 confirmed biological impairment downstream of the Stafford POTW; however, biological impairment was also observed upstream contrary to expectations based on the direction of water flow. A causal analysis using the U.S. Environmental Protection Agency's Stressor Identification Guidance Document helped to identify an illicit, complex episodic toxic discharge. After removal of the illicit discharge, the number of macroinvertebrate taxa increased; however, overall biological condition did not meet state biocriteria. A second causal analysis identified probable causes associated with similar sources scattered throughout the watershed that elevated temperature and altered food resources.

PRESENTATION Headwater Intermittent Stream Workshop (Neaeb) 03/18/2004
Fritz, K M., B R. Johnson, AND D M. Walters. Headwater Intermittent Stream Workshop (Neaeb). Presented at New England Association of Environmental Biologists, Hancock, MA, March 17 - 19, 2004.
Abstract: There is no abstract available for this product. If further information is requested, please refer to the bibliographic citation and contact the person listed under Contact field.

PRESENTATION Large River Biological Assessment Methods Workshop 03/18/2004
Flotemersch, J E. Large River Biological Assessment Methods Workshop. Presented at New England Association of Environmental Biologists, Hancock, MA, March 17-19, 2004.
Abstract: There is no abstract available for this product. If further information is requested, please refer to the bibliographic citation and contact the person listed under Contact field.

PRESENTATION Development of Biological Indicators, Methods and Assessment Techniques for Use in Headwater Intermittent Streams 03/17/2004
FRITZ, K. M. Development of Biological Indicators, Methods and Assessment Techniques for Use in Headwater Intermittent Streams. Presented at New England Association of Environmental Biologists, Hancock, MA, March 17 - 19, 2004.
Abstract: There is no abstract available for this product. If further information is requested, please refer to the bibliographic citation and contact the person listed under Contact field.

PRESENTATION Regional Estimates of the Extent of Non-Indigenous Fish Species in the Surface Waters of the United States 02/05/2004
McCormick, F H., D V. Peck, R M. Hughes, T R. Whittier, L. Kleinsasser, M. P. Dunn, T. P. Simon, AND J M. Lazorchak. Regional Estimates of the Extent of Non-Indigenous Fish Species in the Surface Waters of the United States. Presented at Nonindigenous Species Workshop, Research Triangle Park, NC, February 4-6, 2004.
Abstract: There is no abstract available for this product. If further information is requested, please refer to the bibliographic citation and contact the person listed under Contact field.

PRESENTATION Invasion Dynamics of Red Shiner (Cyprinella Lutrensis) in Southeastern Rivers 02/04/2004
WALTERS, DAVID M., M Blum, AND B Rashleigh. Invasion Dynamics of Red Shiner (Cyprinella Lutrensis) in Southeastern Rivers. Presented at 2nd National EPA Nonindigenous Species Workshop, Research Triangle Park, NC, February 03 - 06, 2004.
Abstract: There is no abstract available for this product. If further information is requested, please refer to the bibliographic citation and contact the person listed under Contact field.

PRESENTATION A Comparison of General Pollution Tolerance Value Lists and Their Use in a Biotic Index 02/04/2004
Winters, L A., K A. Blocksom, F A. Fulk, AND J E. Flotemersch. A Comparison of General Pollution Tolerance Value Lists and Their Use in a Biotic Index. Presented at Western Tolerance Values Workshop, Corvallis, OR, February 2-6, 2004.
Abstract: There is no abstract available for this product. If further information is requested, please refer to the bibliographic citation and contact the person listed under Contact field.

PRESENTATION Development of Biological Indicators, Methods and Assessment Techniques for Use in Headwater Intermittent Streams 02/04/2004
Fritz, K M. Development of Biological Indicators, Methods and Assessment Techniques for Use in Headwater Intermittent Streams. Presented at Surface Water Monitoring and Standards Meeting, Chicago, IL, February 04 - 05, 2004.
Abstract: Despite representing the most abundant and widespread of our nation's surface waters, regions, states and tribes have received little guidance specific to headwater intermittent streams from the U.S. EPA. Headwater streams lie at the terrestrial-aquatic interface both spatially, because of their narrow channel widths and landscape position, and temporally, because of their relatively young geological age and recent transition from terrestrial to aquatic habitats. Perhaps as important, many have physical characteristics of both aquatic and terrestrial habitats because of seasonal flowing and drying phases. These characteristics make the headwater ecosystem a challenging environment for regions, states, and tribes to address regulatory issues.
The Ecological Exposure Research Division (EERD) has received funding through the Regional Methods (RM) Program (sponsored by EPA Regions 1, 2, 3, 4, 5, 9 and 10) to 1) develop standard field protocols for assessing headwater streams (including those that are intermittent); 2) identify appropriate sampling index periods in various regions to maximize biomonitoring potential; 3) identify indicators of hydrologic permanence; and 4) identify potential stressors/impairments to headwater streams.

During the first year (2003) of the study, physical and biological data were collected from 61 sites across a flow permanence gradient and four geographic locations. These locations are Robinson Forest Wildlife Management Area in east central Kentucky, Hoosier National Forest in south central Indiana, Wayne National Forest in south east Ohio, and Edge of Appalachia Nature Preserve in south central Ohio. Preliminary results indicate that channel measures (e.g., bankfull width, channel slope) may be useful for determining the hydrologic permanence of a reach; however, these measures were regionally specific. The slopes of invertebrate species-area relationships appear to vary to a larger extent by region than by hydrologic permanence or habitat type. In addition, total abundance of salamanders did not differ between perennial and intermittent reaches.

The RM study will provide the necessary field protocols for regions, states and tribes to determine the biological integrity and designated uses for the "capillaries" of the river network. The information and tools generated from this study should enable natural resource managers to take advantage of headwater streams as potentially useful study units for investigating the possible effects of land use on water quality. For example, assessment of condition within headwater reaches provides better resolution of land use effects on streams. There is less heterogeneity of land use within smaller basins than larger basins, and therefore, cause of impairment should be more apparent. Additionally, because first order streams are more numerous than higher order streams, replicated studies investigating land use effects should be more feasible. Finally, identifying multiple streams in reference condition is more likely for headwater streams than large water bodies. By building a better understanding of the hydrology associated with headwater streams, these systems can then be useful in our understanding of land use effects on water quality both within small watersheds and across landscapes.

PRESENTATION Applying the Clean Water Act So That We Fix the Right Things 01/23/2004
Cormier, S M., C. Bellucci, AND G. Hoffman. Applying the Clean Water Act So That We Fix the Right Things. Presented at Ohio State University Seminar Series, Columbus, OH, January 23, 2004.
Abstract: Determining the cause of a biological impairment can be a daunting task, one that is complicated by multiple possible causes, spotty or irregularly collected data, natural variations due to season, weather, land formation and other factors, and a lack of basic scientific information needed to diagnose an environmental problem. Nevertheless, probable causes of biological impairments can be determined and findings can be used to reduce or remove detrimental causes and improve condition of biological communities. For example, a 2.4 km segment of the Willimantic River in northeastern Connecticut was listed as impaired in 1998 based on a review of aquatic toxicity monitoring reports submitted to the Connecticut Department of Environmental Protection from the Stafford publicly owned treatment works (POTW). Calculations showed a high probability of exceeding Connecticut's water quality criteria for copper, lead and zinc. Subsequent monitorig in the fall of 1999 confirmed biological impairment downstream of the Stafford POTW, however, biological impairment was also observed upstream contrary to expectations based on the direction of water flow. A causal analysis using the U.S. Environmental Protection Agency's Stressor Identification Document helped to identify an illicit, complex episodic toxic discharge. After removal of the illicit discharge, the number of macroinvertebrate taxa increased; however, overall biological condition did not meet state biocriteria. A second causal analysis identified probable causes associated with similar sources scattered throughout the watershed that elevated temperature and altered food resources.

PUBLISHED REPORT The Presence of Estrogenic and Androgenic Substances in Effluents from Concentrated Animal Feeding Operations 11/01/2004
Lazorchak, J M. AND M E. Smith. The Presence of Estrogenic and Androgenic Substances in Effluents from Concentrated Animal Feeding Operations. U.S. Environmental Protection Agency, Washington, DC, EPA/600/R-04/172, 2004.
Abstract: In February 2003 the U.S.EPA published a final rule on National Polllutant Discharge Elimination System Permit Regulation and Effluent Limitation Guidelines and Standards for Concentrated Animal Feeding Operations (CAFOs). Manure and wastewater from CAFOs have the potential to contribute pollutants such as nitrogen and phosphorus, organic matter, sediments, pathogens, heavy metals, hormones, antibiotics and ammonia to the environment. Excess nutrients in water (e.g., nitrogen and phosphorus) can result in or contribute to low levels of dissolved oxygen (anoxia), eutrophication and toxic algal blooms. The CAFO rule mainly covers the control of nutrients and bacterial contamination. This report investigated the potential ecological effects of hormonally-active substances associated with discharges from cattle feeding operations. Revalor S7 implants, which contain both trenbolone acetate and 17B-estradiol were used on the 6000+ cattle that were housed at the study CAFO facility. Trenbolone acetate is a synthetic anabolic steroid that is widely used in beef production in the U.S. Metabolites of trenbolone acetate include the stereoisomers 17a- and 17B-trenbolone, both of which are stable in animal wastes and are relatively potent androgens in reproduction assays with the fathead minnow, laboratory rat, and other species. The purpose of this study was to evaluate the potential of a fathead minnow vitellogenin gene expression assay as an indicator of androgenic substances like 17a- and 17B-trenbolone in a beef feedlot discharge, and in river water upstream and downstream from the discharge. Our objective was to see if sexually mature female fathead minnows would show reduced levels of vitellogenin gene expression when exposed to androgens. Samples were collected on three different occasions during 2002 and 2003. No consistent changes in vitellogenin gene expression were detected in female fathead minnows exposed to samples collected from a CAFO drain or downstream stream samples that had detectable concentrations of trenbolone shown in laboratory studies to cause effects on reproduction.

PUBLISHED REPORT The Use of Hyperspectral Remote Sensing for the Development of Optical Water Quality Indicators in the Ohio River Basin 11/01/2004
Autrey, B C. The Use of Hyperspectral Remote Sensing for the Development of Optical Water Quality Indicators in the Ohio River Basin. U.S. Environmental Protection Agency, Washington, DC, EPA/600/R-04/170 (NTIS PB2007-106631), 2004.
Abstract: Hyperspectral remote sensing for the assessment of inland water quality can be used in enhancing the capabilities of resource managers to monitor water bodies in a timely and cost-effective manner. The key factor in assessing the accuracy of water quality assessments based on remote sensing is determining the relationships between optical indicators of water quality and remotely sensed data. The usefulness of the optical indicators may depend in large part to their applicability to interpreting data derived from multiple water bodies. The inland water quality parameters have been evaluated using both airborne and satellite-bound sensors. In 1999, a Compact Airborne Spectrographic Imager (CASI) was flown by airplane over the relatively shallow Great Miami River (GMR), Ohio, collecting hyperspectral bands of data. Corresponding water quality samples and field spectrometer data were collected directly from the river during the time of the flight. A similar study was executed in 2001 during which a CASI sensor was flown over a portion of the relatively deep Ohio River while the same types of ground-truth data were collected. Using the remotely sensed, field spectrometer and laboratory analyses data from each of these projects, spectral indices for the analysis of chlorophyll a, turbidity, phosphorus, and nitrogen were developed. The spectral indices developed during the 1999 GMR study were tested against the spectral indices developed for the Ohio River in 2001. While the GMR's spectral index for chlorophyll a was found to be applicable to the Ohio River data, the spectral indices for turbidity, phosphorus and nitrogen were required to be slightly refined in order to be applicable to this new environmental setting.
In 2002 and 2003, hyperspectral data were acquired from the Ohio River using the Hyperion satellite. Near the time that the satellite collected these data, a field crew collected ground-truth data types similar to those collected during the 1999 and 2001 studies. The same spectral index that worked well in predicting chlorophyll a concentrations in the 1999 and 2001 studies were applied to the data collected by the satellite. The results of that application helped validate the chlorophyll-a spectral index.

This study demonstrates the ubiquitous application of the chlorophyll a spectral index while revealing the limited reliability of the turbidity, phosphorous and nitrogen spectral indices. Although differences between the dynamics of the two rivers may have made these spectral indices incompatible, with further refinement they may yet prove to be useful tools that can be modified for use in other rivers to detect potential water quality problems.

PUBLISHED REPORT Vitellogenin Gene Expression in Fathead Minnows and Pearl Dace from Control (Non-Dosed) and Lakes Dosed With Ee2 in the Canadian Experimental Lakes Area 10/01/2004
Flick, R, J M. Lazorchak, AND M E. Smith. Vitellogenin Gene Expression in Fathead Minnows and Pearl Dace from Control (Non-Dosed) and Lakes Dosed With Ee2 in the Canadian Experimental Lakes Area. U.S. Environmental Protection Agency, Washington, DC, EPA/600/R-04/173 (NTIS PB2006-114611), 2004.
Abstract: A whole-lake endocrine disruption experiment was conducted by Fisheries and Oceans Canada at the Experimental Lakes Area (ELA) in northwestern Ontario for three years beginning in 2001. This experiment examined population, organismal, biochemical and cellular-level effects in lake trout, white sucker, fathead minnow and pearl dace exposed to environmentally- relevant (4-6 ng/L) concentrations of the synthetic estrogen, 17a-ethynylestradiol. The USEPA collaborated in this study by evaluating vitellogenin (vtg) gene expression in: 1) indigenous fathead minnows and pearl dace collected from the dosed and control lakes in 2001 through 2003, before and after dosing; 2) indigenous minnows collected in 2001 from the reference lake and deployed for 1, 3, 7 and 13 days in the dosed and reference lakes; and 3) Cincinnati cultured minnows exposed to water collected in 2001 through 2004 from ELA lakes and shipped to Cincinnati. RT-PCR methods were used to measure vtg expression. In addition to water exposures, embryo-larval fish and adult male fathead minnows were exposed to reference and dosed lake sediment elutriates. Indigenous male fathead minnows and pearl dace collected at all time intervals from the dosed lake showed a constant level of elevation in vitellogenin gene expression. Gene expression in the 2001 fathead minnow deployment study was detected within 24 hours after deployment of control fish into the treated lake and stayed elevated for the entire 13-day study. Highly variable gene expression was found in fathead minnow fry exposed to dosed lake sediments elutriates, but no significant gene expression was found in fry exposed to reference lake sediment elutriates. Male adult fathead minnows exposed to elutriates from sediments collected in 2004 in the previously dosed lake showed significant vitellogenin gene expression. Results indicate that RT-PCR analyses of total RNA can be used to provide a rapid and timely estimate of exposure to estrogenic substances.

PUBLISHED REPORT Association Among Invertebrates and Habitat Indicators for Large Rivers in the Midwest 10/01/2004
Flotemersch, J E., K A. Blocksom, J J. Hutchens, AND B C. Autrey. Association Among Invertebrates and Habitat Indicators for Large Rivers in the Midwest. U.S. Environmental Protection Agency, Washington, DC, EPA/600/R-04/177 (NTIS PB2006-114610), 2004.
Abstract: Six reaches in each of two large rivers (one each in Kentucky and Ohio) were sampled using a prototype benthic macroinvertebrate sampling technique. The intent was to better understand the relationship between large river macroinvertebrate assemblages and habitat features. This information was to determine an acceptable sampling design to support development of a large river bioassessment protocol (LR-BP). Specific objectives included determining the appropriate number of habitat point-samples to be collected, examining how varying reach length affects assemblage characteristics, and determining an appropriate laboratory subsample size to accompany the resulting field sampling method. At each site, both banks of 12 transects separated by increasingly larger distances were sampled. Analyses were conducted using Monte Carlo methods. Interpretation of results relied on the metric values of total taxa richness, mayfly taxa richness, caddisfly taxa richness, Diptera richness, % mayflies, % caddisflies, % Tanytarsini, % non-Tanytarsini dipterans and non-insects, and % tolerant individuals.
This research indicates that, using the sampling technique discussed herein, a representative sample of the benthic macroinvertebrate fauna in the study reaches was collected by sampling both banks of 6 transects spaced at 100 m intervals over a 500-m distance. It is hypothesized that these results were achieved because the sampling method and design effectively sampled the benthic macroinvertebrate fauna of the dominant habitat types within the reach.

It is recommended that the field method be coupled with a fixed laboratory subsample size of 300 organisms for bioassessment purposes, with the recognition that a subsample size of 500 organisms may be needed to meet the objectives of some studies. This recommendation is based on the response of the tested metrics, and the observation that the ability to separate sites of different macroinvertebrate composition generally did not increase with larger subsample sizes. It is likely this approach will over-sample sites of uniform composition, but the goal was to develop a standardized LR-BP that would perform well across sites of differing habitat composition. It should be noted that the LR-BP for macroinvertebrates has only been tested in main-channel habitats. It may work equally well on off-channel habitats, but this remains to be tested.

While the method has been designed to perform well in a variety of habitats, resulting data should be interpreted with appreciation for coarse habitat characteristics. This information would ideally be derived from habitat data collected concomitantly with the faunal data. Sites can then be categorized into river types (e.g., impounded vs. free-flowing or lowland vs. upland rivers) or even habitat types within a specific river (e.g., sandy- vs. cobble-bottom) in a more controlled environment (i.e., in the laboratory), thus increasing the overall integrity of any and all site assessments. Possible modifications to the method to streamline its future application in the field are provided.

PUBLISHED REPORT A Review of Biological Assessment Tools and Biocriteria for Streams and Rivers in New England States 10/01/2004
SHELTON, A. D. AND K. A. BLOCKSOM. A Review of Biological Assessment Tools and Biocriteria for Streams and Rivers in New England States. U.S. Environmental Protection Agency, Washington, DC, EPA/600/R-04/168 (NTIS PB2005-109882), 2004.
Abstract: The primary purpose of this document is to serve as a detailed description of the biological assessment programs for wadeable streams and rivers within U.S. Environmental Protection Agency Region 1 states (i.e., Connecticut, Maine, Massachusetts, New Hampshire, Rhode Island and Vermont). Specifically, this report concentrates on the target assemblages (e.g., benthic macroinvertebrates, periphyton and/or fish) and the specific methods used by each state to determine whether biocriteria set for aquatic life use are met in wadeable streams and rivers. The information contained in this report is critical to the eventual use of state data in assessing water resources on a national scale because it provides the necessary level of detail on New England state bioassessment methodologies in a single document. In addition, this report serves as a valuable resource for other states, tribes and municipalities, both those developing bioassessment tools and those with existing programs.

PUBLISHED REPORT Genetic Diversity of Stream Fish in the Mid-Atlantic Integrated Assessment Area 09/30/2004
Bagley, M J., S A. Jackson, S E. Franson, AND E R. Waits. Genetic Diversity of Stream Fish in the Mid-Atlantic Integrated Assessment Area. U.S. Environmental Protection Agency, Washington, DC, EPA/600/R-04/175 (NTIS PB2006-114613), 2004.
Abstract: In this report, we describe the results of research to assess the genetics of three stream fish species in the Mid-Atlantic region. This research had three specific goals. First, we sought to examine the utility of genetic analysis for purposes of taxonomic identification. Bioassessments depend critically on accurate taxonomic identification, yet for many species morphological identification is problematic and prone to error. The use of "DNA taxonomy" may provide an effective solution to improve the accuracy, consistency, and precision of taxonomic identification. Second, we sought to understand the population structure of the three target species within the Mid-Atlantic region. Most species are divided into population segments that may be partially or completely reproductively isolated. These populations represent the fundamental resource units for these species, and therefore are important to delineate when performing ecological assessments. If the population boundaries are similar for different species, then precise biogeographic provinces could be identified using this methodology. Finally, we sought to determine the relationship between measurements of genetic diversity for these three stream fish and stressor profiles previously generated for the Mid-Atlantic region. If a genetic signature of stressors can be detected based on their cumulative impacts on populations, measurements of genetic diversity may provide an effective leading indicator of population level impacts prior to population extirpations.

PUBLISHED REPORT Electrofishing in Boatable Rivers: Does Sampling Design Affect Bioassessment Metrics? 09/30/2004
Flotemersch, J E. AND K A. Blocksom. Electrofishing in Boatable Rivers: Does Sampling Design Affect Bioassessment Metrics? U.S. Environmental Protection Agency, Washington, DC, EPA/600/R-04/174 (NTIS PB2006-114609), 2004.
Abstract: Data were collected from 60 boatable sites using an electrofishing design that permitted comparisons of the effects of designs and distances on fish assemblage metrics. Sites were classified a priori as Run-of-the-River (ROR) or Restricted Flow (RF). Data representing four different design options (i.e., 1000 and 2000 m for both single and paired banks) were extracted from the dataset and analyzed.
Friedman tests comparing metric values among the designs detected significant differences for all richness metrics at both types of sites and for catch per unit effort and percent tolerant species at ROR sites. Richness metrics were generally higher for the two 2000-m designs than for the two 1000-m designs. When plotted against cumulative electrofishing distance, the percent change in metrics declined sharply within approximately 1000 m, after which metrics usually varied by less than 10 percent. These data demonstrate that designs electrofishing 1000 m of shoreline are sufficient for bioassessments on boatable rivers similar to those in this study, regardless of whether the shoreline is along a single bank or distributed equally among paired banks. However, at sites with depths > 4 m, it may be advisable to employ nighttime electrofishing or increase day electrofishing designs to 2000 m.

 

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