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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,

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Presentation on theme: "Do They Eat Petroleum Hydrocarbons? Bioremediation of Toluene by Oil-Water Separator Bacteria LISA L. LOFTIS and JAMES E. CHAMPINE. Biology Department,"— Presentation transcript:

1 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.

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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