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Do They Eat Petroleum Hydrocarbons? Bioremediation of Toluene by Oil-Water Separator Bacteria

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

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  1. 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.

  2. Site History • Former Gasoline Station • Soil and Groundwater Contamination • Why Remediation System was chosen

  3. Site Map

  4. Soil Vapor Extraction Trailer

  5. Hypothesis • Contaminated groundwater represents a natural enrichment for petroleum hydrocarbon degrading bacteria

  6. 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

  7. Site Materials • Before and After groundwater samples • Oil/Water Separator Biofilm (Jar and Vial) • Collection of Bacterial Isolates

  8. Gas Chromatograph

  9. Chromatography Conditions • Carrier Gas • DB1 30 m capillary column • 200ºC oven temperature • Flame Ionization Detector

  10. Typical chromatogram

  11. Serum Bottle Cultures • 20 mL of sterile strength 0.1x TSB • Teflon Rubber Stoppers • Aluminum Crimps • Amended with 10 L of Toluene • Aerobic

  12. Serum Bottle Cultures

  13. Before Groundwater Experiment

  14. After Groundwater Experiment

  15. Isolations • Before • Beige (from After groundwater) • ECP 6 • ECP 7

  16. Toluene Degraders ECP 6 and ECP 7

  17. Conclusions • Intrinsic Bioremediation did occur at the site • Site cleaned up in half of the estimated time • Bacteria with toluene degradation activity were isolated

  18. Acknowledgments • Southeast Missouri State University Undergraduate Research Program • Heather DuShell • Dr. Bjorn Olesen and the Department of Chemistry • Vicki Howell, Biology Dept. Secretary

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