Microbial characterization of a JP-4 fuel-contaminated site using a combined lipid biomarker/polymerase chain reaction–denaturing gradient gel electrophoresis (PCR–DGGE)-based approach

Authors

  • John R. Stephen,

    1. Center for Environmental Biotechnology, The University of Tennessee, 10515 Research Drive, Suite 300, Knoxville, TN 37932, USA.,
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  • Yun-Juan Chang,

    1. Center for Environmental Biotechnology, The University of Tennessee, 10515 Research Drive, Suite 300, Knoxville, TN 37932, USA.,
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  • Ying Dong Gan,

    1. Center for Environmental Biotechnology, The University of Tennessee, 10515 Research Drive, Suite 300, Knoxville, TN 37932, USA.,
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  • Aaron Peacock,

    1. Center for Environmental Biotechnology, The University of Tennessee, 10515 Research Drive, Suite 300, Knoxville, TN 37932, USA.,
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  • Susan M. Pfiffner,

    1. Center for Environmental Biotechnology, The University of Tennessee, 10515 Research Drive, Suite 300, Knoxville, TN 37932, USA.,
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  • Michael J. Barcelona,

    1. National Center for Integrated Bioremediation Research and Development, Department of Civil and Environmental Engineering, University of Michigan, MI 48109, USA.,
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  • David C. White,

    1. Center for Environmental Biotechnology, The University of Tennessee, 10515 Research Drive, Suite 300, Knoxville, TN 37932, USA.,
    2. Environmental Sciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA.
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  • Sarah J. Macnaughton

    1. Center for Environmental Biotechnology, The University of Tennessee, 10515 Research Drive, Suite 300, Knoxville, TN 37932, USA.,
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Sarah J. Macnaughton E-mail sjmac@utk.edu; Tel. (+1) 423 974 8005; Fax (+1) 423 974 8027.

Abstract

The impact of pollution on soil microbial communities and subsequent bioremediation can be measured quantitatively in situ using direct, non-culture- dependent techniques. Such techniques have advantages over culture-based methods, which often account for less than 1% of the extant microbial community. In 1988, a JP-4 fuel spill contaminated the glacio-fluvial aquifer at Wurtsmith Air Force Base, Michigan, USA. In this study, lipid biomarker characterization of the bacterial and eukaryotic communities was combined with polymerase chain reaction– denaturing gradient gel electrophoresis (PCR–DGGE) analysis of the eubacterial community to evaluate correlation between contaminant (JP-4 fuel) concentration and community structure shifts. Vadose, capillary fringe and saturated zone samples were taken from cores within and up- and down-gradient from the contaminant plume. Lipid biomarker analysis indicated that samples from within the plume contained increased biomass, with large proportions of typically Gram-negative bacteria. Outside the plume, lipid profiles indicated low-biomass microbial communities compared with those within the initial spill site. 16S rDNA sequences derived from DGGE profiles from within the initial spill site suggested dominance of the eubacterial community by a limited number of phylogenetically diverse organisms. Used in tandem with pollutant quantification, these molecular techniques should facilitate significant improvements over current assessment procedures for the determination of remediation end-points.

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