Download - Do They Eat Petroleum Hydrocarbons? Bioremediation of Toluene by Oil-Water Separator Bacteria
Do They Eat Petroleum Do They Eat Petroleum Hydrocarbons? Hydrocarbons? Bioremediation of Toluene Bioremediation of Toluene by Oil-Water Separator by Oil-Water Separator BacteriaBacteria LISA L. LOFTIS and JAMES E. CHAMPINE. LISA L. LOFTIS and JAMES E. CHAMPINE. Biology Department, SoutheastBiology Department, Southeast Missouri State Missouri State University.University.
Site HistorySite History Former Gasoline StationFormer Gasoline Station Soil and Groundwater ContaminationSoil and Groundwater Contamination Why Remediation System was chosenWhy Remediation System was chosen
Site MapSite Map
Soil Vapor Extraction TrailerSoil Vapor Extraction Trailer
HypothesisHypothesis Contaminated groundwater represents a Contaminated groundwater represents a
natural enrichment for petroleum natural enrichment for petroleum hydrocarbon degrading bacteriahydrocarbon degrading bacteria
ApproachApproach Use toluene as a surrogate for petroleum Use toluene as a surrogate for petroleum
related hydrocarbons that may be related hydrocarbons that may be degraded in the aquiferdegraded in the aquifer
Test groundwater biofilm and pure Test groundwater biofilm and pure cultures from the site for toluene cultures from the site for toluene degradation activitydegradation activity
Site MaterialsSite Materials Before and After Before and After
groundwater samples groundwater samples Oil/Water Separator Oil/Water Separator
Biofilm (Jar and Vial)Biofilm (Jar and Vial) Collection of Collection of
Bacterial IsolatesBacterial Isolates
Gas ChromatographGas Chromatograph
Chromatography Chromatography ConditionsConditions
Carrier GasCarrier Gas DB1 30 m capillary columnDB1 30 m capillary column 200200ºC oven temperatureºC oven temperature Flame Ionization DetectorFlame Ionization Detector
Typical chromatogramTypical chromatogram
Serum Bottle CulturesSerum Bottle Cultures 20 mL of sterile 20 mL of sterile
strength 0.1x TSBstrength 0.1x TSB Teflon Rubber Teflon Rubber
StoppersStoppers Aluminum CrimpsAluminum Crimps Amended with 10 Amended with 10
L of TolueneL of Toluene AerobicAerobic
Serum Bottle CulturesSerum Bottle Cultures
Before Groundwater Before Groundwater ExperimentExperimentBefore Groundwater
010000000200000003000000040000000
Date
Area
Und
er C
urve
After Groundwater After Groundwater ExperimentExperiment
After Groundwater (red bacteria)
0200000400000600000800000
1000000
Date
Area
Und
er C
urve
IsolationsIsolations Before Before Beige (from After Beige (from After
groundwater)groundwater) ECP 6ECP 6 ECP 7ECP 7
Toluene Degraders Toluene Degraders ECP 6 and ECP 7ECP 6 and ECP 7
ECP 6 and ECP 7
020000000400000006000000080000000
100000000
Area
Und
er C
urve
Series2Series1
ConclusionsConclusions Intrinsic Bioremediation did occur at the Intrinsic Bioremediation did occur at the
sitesite Site cleaned up in half of the estimated Site cleaned up in half of the estimated
timetime Bacteria with toluene degradation activity Bacteria with toluene degradation activity
were isolatedwere isolated
AcknowledgmentsAcknowledgments Southeast Missouri State University Southeast Missouri State University
Undergraduate Research ProgramUndergraduate Research Program Heather DuShellHeather DuShell Dr. Bjorn Olesen and the Department of Dr. Bjorn Olesen and the Department of
ChemistryChemistry Vicki Howell, Biology Dept. Secretary Vicki Howell, Biology Dept. Secretary