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Cell-based metabolomics for assessing chemical exposure and toxicity of environmental surface waters (presentation)
Teng, Q., D. Ekman, Dan Villeneuve, J. Cavallin, J. Davis, G. Ankley, AND Tim Collette. Cell-based metabolomics for assessing chemical exposure and toxicity of environmental surface waters (presentation). Presented at 11th International Conference of the Metabolomics Society, San Francisco, CA, June 29 - July 02, 2015.
Poster presented at the 11th International Conference of the Metabolomics Society, San Francisco, CA, June 29 to July 2, 2015.
Introduction: Waste water treatment plants (WWTPs), concentrated animal feeding operations (CAFOs), mining activities, and agricultural operations release contaminants that negatively affect surface water quality. Traditional methods using live animals (e.g. fish) to monitor/assess contaminant exposure and impacts in affected ecosystems are both resource and time intensive and are not sustainable in the long term. This study describes an optimized cell culture-based method employing metabolomics combined with high throughput NMR spectroscopic analysis to obtain multi-endpoint cellular responses to stressor exposure. We report results of applying cell culture-based metabolomics using zebrafish (Danio rerio) liver cells (ZFL) to determine impacted biochemical processes and putative biomarkers of exposure to surface waters from contaminated and pristine sites.Methods: Several surface waters were collected at a Great Lakes Area of Concern (AOC), specifically the St. Louis River AOC into which the Western Lake Superior Sanitary District WWTP flows. ZFL cells were then exposed to culture media prepared with the surface water samples using different doses and durations as well as media prepared from water collected at a reference site and using purified laboratory water. The polar fractions of cellular extracts were analyzed by flow NMR automation. Cellular responses were determined by evaluating changes in endogenous metabolite profiles using a combination of PCA and t-test filtered difference spectra.Preliminary Results: PCA analysis indicated that metabolic changes were time- and dose-dependent for the ZFL cells exposed to the environmental surface waters collected at two WWTPs and downstream sites. However, no significant changes were observed after exposure to the surface water from the pristine site, compared to the control water (Lake Superior water). The t-test filtered difference spectra determined that metabolites involved in energy production varied as a function of the exposure concentration and duration. The other primary responses observed were impacts on lipid and glutathione metabolism.Novel Aspects: Cell-based metabolomics is applied to field studies.
Record Details:Record Type: DOCUMENT (PRESENTATION/SLIDE)
Organization:U.S. ENVIRONMENTAL PROTECTION AGENCY
OFFICE OF RESEARCH AND DEVELOPMENT
NATIONAL EXPOSURE RESEARCH LABORATORY
ECOSYSTEMS RESEARCH DIVISION