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DNA DAMAGE AND VITELLOGENIN RESPONSES IN FERAL FISH EXPOSED TO URBANIZATION AND WASTE WATER TREATMENT EFFLUENT IN SOUTH CAROLINA, USA
OTTER, R., D. A. GORDON, J. R. MEIER, AND S. KLAINE. DNA DAMAGE AND VITELLOGENIN RESPONSES IN FERAL FISH EXPOSED TO URBANIZATION AND WASTE WATER TREATMENT EFFLUENT IN SOUTH CAROLINA, USA. Presented at SETAC-Europe, Hague, NETHERLANDS, May 07 - 11, 2006.
This study was designed to look at the impact of urbanization and a wastewater treatment plant (WWTP) by using field-collected bluegill (Lepomis machrochkrus). Fish were collected from four locations in the same river: above city (reference); below city and above the WWTP; directly below the WWTP; and three km downstream of the WWTP and below an impoundment. Results show a significant increase in DNA damage in blood cells (comet assay) below urbanized areas. This effect was reduced downstream of the WWTP, however DNA damage increased at the site furthest downstream, suggesting another stressor source. Juvenile fish collected directly below a WWTP outfall showed a 12-fold induction in vitellogenin (Vg) gene expression over reference fish. Vg expressions in fish collected 3 km downstream of the WWTP were similar to reference fish. Habitat quality assessments, along with chemical concentrations of polyaromatic hydrocarbons in sediments and synthetic estrogens and alkyphenols concentrations in water, showed that habitat quality between all sites was similar and chemical stressors could in part explain the biological effects seen in bluegill.
The indeterminate condition of exposure indicator research stands to change markedly with the ability to connect molecular biological technologies with cellular or tissue effects and outcomes. Three focal areas of ecological research aim to develop a sequence of approaches where "the earliest recognizable signatures of exposure" (i.e., unique patterns of up- and down-regulated genes and proteins) are identified for numerous stressors, demonstrable in case studies and incorporated into Agency, State and Regional studies supported by EMAP and other programs.
Area 1, Computational Toxicology Research: Exposure assessment has historically been based on use of chemical analysis data to generate exposure models. While biological activity of chemicals has been recognized to be important for exposure risk assessments, measurement of such activity has been limited to whole organism toxicity tests. Use of molecular approaches will:
improve extrapolation between components of source-to-outcome continuum (source , exposure , dose , effect , outcome)
Using a systems modeling approach, gene and protein expression data, in small fish models (fathead minnow and zebrafish), will be integrated with metabolomic and histopathological data. This will assist in prediction of environmental transformation and chemical effects based on structural characteristics, and enhance quantitative risk assessments, including areas of uncertainty such as a basis for extrapolation of effects of endocrine disrupting chemicals, interspecies extrapolation, complex chemical mixtures and dose-response assessment.
Area 2, Ecological Research-Environmental Diagnostics: Development of molecular diagnostic indicators contributes to several of the GPRA Diagnostic Research Goals. Methods will employ DNA microarray technology and expression proteomics, focusing on species of relevance to aquatic ecosystem risk assessment. Significantly, these diagnostic indicators will open the door to understanding subcellular interactions resulting from exposure to complex chemical mixtures.
define relationship between genetic disposition of populations and degree/specificity of stressor-specific gene transcriptional response in aquatic organisms (fish and invertebrates)
identify of chemical mixture induced transcriptional "patterns" using microarrays and hyperspectral scanning - via collaboration with DOE Sandia National Labs
apply molecular indicators to watershed level stressor study, including pilot studies with targeted pesticides and toxins indicators
develop molecular indicators of exposure for invertebrates (Daphnia, Lumbriculus, Chironomus)
Area 3, Exposure Research in Endocrine Disruptors:
Subobjective 1: Develop exposure methods, measurement protocols, and models for assessment of risk management practices of endocrine disrupting compounds. As risk management approaches are identified and developed, there will be a need to identify, adapt and develop bioassay screening tools and other analytical methods to assess their efficacy. Measurements research will be performed to define management needs. This effort will entail cross-lab participation from NRMRL, NERL and NHEERL.
Subobjective 2: Determine extent of environmental and human exposures to EDCs, characterize sources and factors influencing these exposures, develop and evaluate risk management strategies to reduce exposures. In order to develop effective risk management strategies, it is important to understand the extent of exposures to endocrine disrupting compounds and factors influencing source-to-exposure-to-dose relationships.
apply molecular indicators of exposure to estrogenic compounds in selected wastewater treatment plants located in ten USEPA Regions
identify differential gene expression following exposure of fathead minnows to environmental androgens and androgen-like compounds
apply molecular indicators of exposu
Record Details:Record Type: DOCUMENT (PRESENTATION/ABSTRACT)
Organization:U.S. ENVIRONMENTAL PROTECTION AGENCY
OFFICE OF RESEARCH AND DEVELOPMENT
NATIONAL EXPOSURE RESEARCH LAB
ECOLOGICAL EXPOSURE RESEARCH DIVISION
MOLECULAR INDICATORS RESEARCH BRANCH