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Do They Eat Petroleum Hydrocarbons? Bioremediation of Toluene by Oil-Water Separator Bacteria. LISA L. LOFTIS and JAMES E. CHAMPINE. Biology Department, Southeast Missouri State University. Site History. Former Gasoline Station Soil and Groundwater Contamination
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Do They Eat Petroleum Hydrocarbons? Bioremediation of Toluene by Oil-Water Separator Bacteria LISA L. LOFTIS and JAMES E. CHAMPINE. Biology Department, SoutheastMissouri State University.
Site History • Former Gasoline Station • Soil and Groundwater Contamination • Why Remediation System was chosen
Hypothesis • Contaminated groundwater represents a natural enrichment for petroleum hydrocarbon degrading bacteria
Approach • Use toluene as a surrogate for petroleum related hydrocarbons that may be degraded in the aquifer • Test groundwater biofilm and pure cultures from the site for toluene degradation activity
Site Materials • Before and After groundwater samples • Oil/Water Separator Biofilm (Jar and Vial) • Collection of Bacterial Isolates
Chromatography Conditions • Carrier Gas • DB1 30 m capillary column • 200ºC oven temperature • Flame Ionization Detector
Serum Bottle Cultures • 20 mL of sterile strength 0.1x TSB • Teflon Rubber Stoppers • Aluminum Crimps • Amended with 10 L of Toluene • Aerobic
Isolations • Before • Beige (from After groundwater) • ECP 6 • ECP 7
Conclusions • Intrinsic Bioremediation did occur at the site • Site cleaned up in half of the estimated time • Bacteria with toluene degradation activity were isolated
Acknowledgments • Southeast Missouri State University Undergraduate Research Program • Heather DuShell • Dr. Bjorn Olesen and the Department of Chemistry • Vicki Howell, Biology Dept. Secretary