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Applied and Environmental Microbiology, July 2001, p. 3149-3160, Vol. 67, No. 7
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.7.3149-3160.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Diversity and Characterization of Sulfate-Reducing Bacteria in Groundwater at a Uranium Mill Tailings Site

Yun-Juan Chang,1 Aaron D. Peacock,1 Philip E. Long,2 John R. Stephen,3 James P. McKinley,2 Sarah J. Macnaughton,4 A. K. M. Anwar Hussain,1 Arnold M. Saxton,5 and David C. White1,*

Center for Biomarker Analysis1 and Department of Animal Science,5 The University of Tennessee, Knoxville, Tennessee 37932-2575; Environmental Technology, Pacific Northwest National Laboratory, Richland, Washington 993522; and Crop and Weed Science, Horticulture Research International, Wellesbourne, Warwick CV35 9EF,3 and AEA Technology Environment, Abingdon, OX 14 3BD,4 United Kingdom

Received 5 February 2001/Accepted 26 March 2001

Microbially mediated reduction and immobilization of U(VI) to U(IV) plays a role in both natural attenuation and accelerated bioremediation of uranium-contaminated sites. To realize bioremediation potential and accurately predict natural attenuation, it is important to first understand the microbial diversity of such sites. In this paper, the distribution of sulfate-reducing bacteria (SRB) in contaminated groundwater associated with a uranium mill tailings disposal site at Shiprock, N.Mex., was investigated. Two culture-independent analyses were employed: sequencing of clone libraries of PCR-amplified dissimilatory sulfite reductase (DSR) gene fragments and phospholipid fatty acid (PLFA) biomarker analysis. A remarkable diversity among the DSR sequences was revealed, including sequences from delta -Proteobacteria, gram-positive organisms, and the Nitrospira division. PLFA analysis detected at least 52 different mid-chain-branched saturate PLFA and included a high proportion of 10me16:0. Desulfotomaculum and Desulfotomaculum-like sequences were the most dominant DSR genes detected. Those belonging to SRB within delta -Proteobacteria were mainly recovered from low-uranium (<= 302 ppb) samples. One Desulfotomaculum-like sequence cluster overwhelmingly dominated high-U (>1,500 ppb) sites. Logistic regression showed a significant influence of uranium concentration over the dominance of this cluster of sequences (P = 0.0001). This strong association indicates that Desulfotomaculum has remarkable tolerance and adaptation to high levels of uranium and suggests the organism's possible involvement in natural attenuation of uranium. The in situ activity level of Desulfotomaculum in uranium-contaminated environments and its comparison to the activities of other SRB and other functional groups should be an important area for future research.

* Corresponding author. Mailing address: Center for Biomarker Analysis, The University of Tennessee, 10515 Research Dr., Suite 300, Knoxville, TN 37932-2575. Phone: (865) 974-8001. Fax: (865) 974-8027. E-mail: Milipids{at}aol.com

Applied and Environmental Microbiology, July 2001, p. 3149-3160, Vol. 67, No. 7
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.7.3149-3160.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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