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Applied and Environmental Microbiology, April 2001, p. 1542-1550, Vol. 67, No. 4
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.4.1542-1550.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Succession of Phenotypic, Genotypic, and Metabolic Community Characteristics during In Vitro Bioslurry Treatment of Polycyclic Aromatic Hydrocarbon-Contaminated Sediments

David B. Ringelberg,1 Jeffrey W. Talley,1,* Edward J. Perkins,1 Samuel G. Tucker,1 Richard G. Luthy,2 Edward J. Bouwer,3 and Herbert L. Fredrickson1

Environmental Laboratory, U.S. Army Engineer Research and Development Center, Vicksburg, Mississippi 391801; Department of Civil and Environmental Engineering, Stanford University, Stanford, California 94305-40202; and Department of Geography and Environmental Engineering, Johns Hopkins University, Baltimore, Maryland 212183

Received 26 September 2000/Accepted 8 January 2001

Dredged harbor sediment contaminated with polycyclic aromatic hydrocarbons (PAHs) was removed from the Milwaukee Confined Disposal Facility and examined for in situ biodegradative capacity. Molecular techniques were used to determine the successional characteristics of the indigenous microbiota during a 4-month bioslurry evaluation. Ester-linked phospholipid fatty acids (PLFA), multiplex PCR of targeted genes, and radiorespirometry techniques were used to define in situ microbial phenotypic, genotypic, and metabolic responses, respectively. Soxhlet extractions revealed a loss in total PAH concentrations of 52%. Individual PAHs showed reductions as great as 75% (i.e., acenapthene and fluorene). Rates of 14C-PAH mineralization (percent/day) were greatest for phenanthrene, followed by pyrene and then chrysene. There was no mineralization capacity for benzo[a]pyrene. Ester-linked phospholipid fatty acid analysis revealed a threefold increase in total microbial biomass and a dynamic microbial community composition that showed a strong correlation with observed changes in the PAH chemistry (canonical r2 of 0.999). Nucleic acid analyses showed copies of genes encoding PAH-degrading enzymes (extradiol dioxygenases, hydroxylases, and meta-cleavage enzymes) to increase by as much as 4 orders of magnitude. Shifts in gene copy numbers showed strong correlations with shifts in specific subsets of the extant microbial community. Specifically, declines in the concentrations of three-ring PAH moieties (i.e., phenanthrene) correlated with PLFA indicative of certain gram-negative bacteria (i.e., Rhodococcus spp. and/or actinomycetes) and genes encoding for naphthalene-, biphenyl-, and catechol-2,3-dioxygenase degradative enzymes. The results of this study suggest that the intrinsic biodegradative potential of an environmental site can be derived from the polyphasic characterization of the in situ microbial community.

* Corresponding author. Mailing address: U.S. Army Environmental Research and Development Center, 3909 Halls Ferry Rd., Vicksburg, MS 39180-6199. Phone: (601) 634-2856. Fax: (601) 634-4844. E-mail: talleyj{at}wes.army.mil.

Applied and Environmental Microbiology, April 2001, p. 1542-1550, Vol. 67, No. 4
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.4.1542-1550.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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