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2002 Progress Report: Continuation of an Investigation into the Anaerobic Intrinsic Bioremediation of Whole Gasoline
EPA Grant Number: R827015C017Subproject: this is subproject number 017 , established and managed by the Center Director under grant R827015
(EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
Center: IPEC University of Tulsa (TU)
Center Director: Sublette, Kerry L.
Title: Continuation of an Investigation into the Anaerobic Intrinsic Bioremediation of Whole Gasoline
Investigators: Suflita, Joseph , Townsend, G. Todd
Institution: University of Oklahoma
EPA Project Officer: Krishnan, Bala S.
Project Period: March 1, 2001 through February 28, 2002 (Extended to August 28, 2002)
Project Period Covered by this Report: March 1, 2001 through February 28, 2002
RFA: Integrated Petroleum Environmental Consortium (IPEC) (1999)
Research Category: Hazardous Waste/Remediation , Targeted Research
Description:
Objective:The primary objective of this project is to investigate the ability of microorganisms from contaminated aquifers to degrade a variety of hydrocarbon molecules under different electron accepting conditions. Specifically, the degradation of several alicyclic compounds with either sulfate or carbon dioxide as electron acceptor is examined. In addition, the ability of hydrocarbons in several crude oils to stimulate sulfate reduction and methanogenesis is examined. Compounds contained in the oils are noted for their relative susceptibility to anaerobic microbial decay and the preferred electron acceptor similarly examined.
Progress Summary:The biodegradation of petroleum hydrocarbons by anoxic aquifer sediments previously contaminated by natural gas condensate was examined under methanogenic and sulfatereducing conditions. To explore the substrate range of the resident microbiota, two crude oils, rich in high molecular weight constituents, were chosen as substrates: artificially weathered Alaska North Slope crude, an oil that is depleted in low molecular weight alkanes and monoaromatic compounds but retains its normal complement of mid-toheavy range hydrocarbons (C15-C34), and Alba crude oil, an oil that is naturally depleted in n-alkanes of all sizes but retains most of its polycyclic aromatic hydrocarbons. We examined in detail the change in crude oil composition over a 400+ day incubation period to determine the resident microbiota's substrate preference when presented with a petroleum hydrocarbon mixture and the role of sulfate in the biodegradation of individual hydrocarbon components.
Weathered Alaska North Slope oil amendments resulted in a drastic increase in both methanogenesis and sulfate reduction. Alba crude oil amended incubations displayed a relatively modest stimulation of methanogenesis and sulfate reduction (Figure 1).
Figure 1. Cumulative methanogenesis and sulfate reduction in incubations amended with weathered Alaska North Slope Crude (LI) and Alba (D) compared to autoclaved (FI) and unamended (-] ) control incubations.
Hydrocarbon analysis revealed that the n-alkane fraction of the weathered Alaska North Slope crude, consisting predominately of hydrocarbons fourteen to thirty-four carbons in length, was completely biodegraded under both sulfate-reducing and methanogenic conditions after thirteen months of incubation (Figure 2). Unlike the unbranched nalkanes, the branched isoprenoid alkanes pristine and phytane were not biodegraded under either condition. The time course for degradation of two representative alkanes, heptadecane (C17H36) and triacontane (C30IL62), is also given in Figure 2. As illustrated by these examples, the entire range of n-alkanes was consumed simultaneously with only slightly reduced rates for the longer chain congeners. The n-alkanes were biodegraded approximately twice as fast in the presence of sulfate as they were in its absence. Nevertheless, by the end of the incubation period, all unsubstituted n-alkanes originating from the weathered Alaska North Slope crude were completely consumed under both conditions.
Figure 2. n-Alkane biodegradation in weathered Alaska North Slope crude oil. (A) Chromatograms of residual oil after 13 months of incubation. (B) Time course of biodegradation of two n-alkanes.
Polycyclic aromatic hydrocarbons on the USEPA priority pollutant list were also examined. In sulfate reducing incubations amended with either oil, naphthalene, 2methylnaphthalene, and 2-ethyinaphthalene were biodegraded in that order of preference (Figure 2). In incubations amended with Alba oil, the coeluting 2,6-and 2,7dimethylnaphthalenes were also biodegraded. The I -methyl- and I -ethyl-naphthalene congeners as well as the dimethyl-substituted naphthalenes within these incubations were recalcitrant. No biodegradation of naphthalene or its alkylated congeners was seen in incubations held under methanogenic conditions. We believe this is the first report of 2ethyinaphthalene and of 2,6- and 2,7-dimetylynaphthalene biodegradation under anaerobic conditions
Figure 3. Selective biodegradation of naphthalene, 2-methylnaphthalene, and 2ethyinaphthalene in crude oil amended incubations held under sulfate reducing conditions after 14 months.
These results indicate that n-alkanes, regardless of their size, are relatively labile in anaerobic compartments and their biodegradation in terrestrial environments is not limited by electron acceptor availability. Polycyclic aromatic hydrocarbons, on the other hand, are much more recalcitrant, and their biodegradation involves biodegradative activities that are both sulfate-dependent and congener-specific. This information should be useful in judging the extent of in situ biodegradation of crude oil in terrestrial envirorunents and in making decisions regarding risk-based corrective actions.
We report that these aquifer sediments harbor microorganisms are able to biodegrade a wide range of high molecular weight alkanes, even though they are not significant components of the native contamination, under both sulfate-reducing and methanogenic conditions. Selected naphthalenes were also biodegraded exclusively under sulfatereducing conditions. These results demonstrate the presence of diverse petroleum biodegradative activities in anaerobic terrestrial environments that transcends previous hydrocarbon exposure. This study, which took advantage of complex hydrocarbon mixtures as substrate and unenriched microbial communities as inoculum, shed light on the extent to which in situ biodegradative processes are acting upon residual contamination and hopefully will influence decisions regarding the need for remediation.
Journal Articles on this Report: 2 Displayed | Download in RIS Format
| Other subproject views: | All 2 publications | 2 publications in selected types | All 2 journal articles |
| Other center views: | All 135 publications | 26 publications in selected types | All 19 journal articles |
| Type | Citation | ||
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Rios-Hernandez, L. A., Gieg, L. M., and Suflita, J. M. Biodegradation of an alicyclic hydrocarbon by a sulfate-reducing enrichment from a gas condensate-contaminated aquifer. (2003) Applied and Environmental Mirobiology, 69, (1), 434-443. |
R827015C017 (2002) R827015C017 (Final) |
not available |
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Townsend GT, Prince RC, Suflita JM (2003) Anaerobic oxidation of crude oil hydrocarbons by the resident microorganisms of a contaminated anoxic aquifer. Environmental Science and Technology 37:5213-5218 |
R827015C004 (2001) R827015C017 (2002) R827015C017 (Final) |
not available |
groundwater, hydrocarbons, biodegradation, alicyclic, alkanes, sulfate reduction, methanogenesis, ethylcyclopentane, anaerobic, community analysis, denaturing gradient gel electrophoresis, gas chromatography/mass spectrometry (GCMS), PAH'S, bacteria, bioremediation, pathway, intermediates, microbial ecology, aquifer, oil, gas condensate, contamination, gasoline, fumarate addition. , Ecosystem Protection/Environmental Exposure & Risk, POLLUTANTS/TOXICS, INTERNATIONAL COOPERATION, Geographic Area, TREATMENT/CONTROL, Scientific Discipline, Waste, RFA, Remediation, Biology, Civil/Environmental Engineering, Northwest, Microbiology, Bioavailability, Chemicals, Chemistry, Hazardous Waste, Biochemistry, Environmental Engineering, Environmental Microbiology, Groundwater remediation, Hazardous, Oil Spills, Treatment Technologies, Bioremediation, Engineering, State, risk assessment, gasoline, anaerobic bioconversion, biodegradation, hydrocarbons, anaerobic biodegradation, anaerobic bioremediation, Ft. Lupton, CO, groundwater, risk assessments, electron acceptor, petrochemical waste, biological markers, petroleum contaminants, intrinsic bioremediation, hazardous waste treatment, BTEX
Progress and Final Reports:
Original Abstract
Final Report
Main Center Abstract and Reports:
R827015 IPEC University of Tulsa (TU)
Subprojects under this Center:
(EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
R827015C001 Evaluation of Road Base Material Derived from Tank Bottom Sludges
R827015C002 Passive Sampling Devices (PSDs) for Bioavailability Screening of Soils Containing Petrochemicals
R827015C003 Demonstration of a Subsurface Drainage System for the Remediation of Brine-Impacted Soil
R827015C004 Anaerobic Intrinsic Bioremediation of Whole Gasoline
R827015C005 Microflora Involved in Phytoremediation of Polyaromatic Hydrocarbons
R827015C006 Microbial Treatment of Naturally Occurring Radioactive Material (NORM)
R827015C007 Using Plants to Remediate Petroleum-Contaminated Soil
R827015C008 The Use of Nitrate for the Control of Sulfide Formation in Oklahoma Oil Fields
R827015C009 Surfactant-Enhanced Treatment of Oil-Contaminated Soils and Oil-Based Drill Cuttings
R827015C010 Novel Materials for Facile Separation of Petroleum Products from Aqueous Mixtures Via Magnetic Filtration
R827015C011 Development of Relevant Ecological Screening Criteria (RESC) for Petroleum Hydrocarbon-Contaminated Exploration and Production Sites
R827015C012 Humate-Induced Remediation of Petroleum Contaminated Surface Soils
R827015C013 New Process for Plugging Abandoned Wells
R827015C014 Enhancement of Microbial Sulfate Reduction for the Remediation of Hydrocarbon Contaminated Aquifers - A Laboratory and Field Scale Demonstration
R827015C015 Locating Oil-Water Interfaces in Process Vessels
R827015C016 Remediation of Brine Spills with Hay
R827015C017 Continuation of an Investigation into the Anaerobic Intrinsic Bioremediation of Whole Gasoline
R827015C018 Using Plants to Remediate Petroleum-Contaminated Soil
R827015C019 Biodegradation of Petroleum Hydrocarbons in Salt-Impacted Soil by Native Halophiles or Halotolerants and Strategies for Enhanced Degradation
R827015C020 Anaerobic Intrinsic Bioremediation of MTBE
R827015C021 Evaluation of Commercial, Microbial-Based Products to Treat Paraffin Deposition in Tank Bottoms and Oil Production Equipment
R827015C022 A Continuation: Humate-Induced Remediation of Petroleum Contaminated Surface Soils
R827015C023 Data for Design of Vapor Recovery Units for Crude Oil Stock Tank Emissions
R827015C024 Development of an Environmentally Friendly and Economical Process for Plugging Abandoned Wells
R827015C025 A Continuation of Remediation of Brine Spills with Hay
R827015C026 Identifying the Signature of the Natural Attenuation of MTBE in Goundwater Using Molecular Methods and "Bug Traps"
R827015C027 Identifying the Signature of Natural Attenuation in the Microbial
Ecology of Hydrocarbon Contaminated Groundwater Using Molecular Methods and
"Bug Traps"
R827015C028 Using Plants to Remediate Petroleum-Contaminated Soil: Project Continuation
R827015C030 Effective Stormwater and Sediment Control During Pipeline Construction Using a New Filter Fence Concept
R827015C031 Evaluation of Sub-micellar Synthetic Surfactants versus Biosurfactants for Enhanced LNAPL Recovery
R827015C032 Utilization of the Carbon and Hydrogen Isotopic Composition of Individual Compounds in Refined Hydrocarbon Products To Monitor Their Fate in the Environment
R830633 Integrated Petroleum Environmental Consortium (IPEC)
R830633C001 Development of an Environmentally Friendly and Economical Process for Plugging Abandoned Wells (Phase II)
R830633C002 A Continuation of Remediation of Brine Spills with Hay
R830633C003 Effective Stormwater and Sediment Control During Pipeline Construction Using a New Filter Fence Concept
R830633C004 Evaluation of Sub-micellar Synthetic Surfactants versus Biosurfactants for Enhanced LNAPL Recovery
R830633C005 Utilization of the Carbon and Hydrogen Isotopic Composition of Individual Compounds in Refined Hydrocarbon Products To Monitor Their Fate in the Environment
R830633C006 Evaluation of Commercial, Microbial-Based Products to Treat Paraffin Deposition in Tank Bottoms and Oil Production Equipment
R830633C007 Identifying the Signature of the Natural Attenuation in the Microbial Ecology of Hydrocarbon Contaminated Groundwater Using Molecular Methods and “Bug Traps”
R830633C008 Using Plants to Remediate Petroleum-Contaminated Soil: Project Continuation
R830633C009 Use of Earthworms to Accelerate the Restoration of Oil and Brine Impacted Sites
X832428C001 Effective Stormwater and Sediment Control During Pipeline Construction Using a New Filter Fence Concept
X832428C002 Paraffin Control in Oil Wells Using Anaerobic Microorganisms
X832428C003 Fiber Rolls as a Tool for Re-Vegetation of Oil-Brine Contaminated Watersheds