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Applied and Environmental Microbiology, March 2003, p. 1337-1346, Vol. 69, No. 3
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.3.1337-1346.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Microbial Populations Stimulated for Hexavalent Uranium Reduction in Uranium Mine Sediment

Yohey Suzuki,1* Shelly D. Kelly,2 Kenneth M. Kemner,2 and Jillian F. Banfield3

Department of Geology and Geophysics, University of Wisconsin—Madison, Madison, Wisconsin 53706,1 Environmental Research Division, Argonne National Laboratory, Argonne, Illinois 60439,2 Department of Earth and Planetary Sciences, University of California—Berkeley, Berkeley, California 947203

Received 23 May 2002/ Accepted 6 September 2002

Uranium-contaminated sediment and water collected from an inactive uranium mine were incubated anaerobically with organic substrates. Stimulated microbial populations removed U almost entirely from solution within 1 month. X-ray absorption near-edge structure analysis showed that U(VI) was reduced to U(IV) during the incubation. Observations by transmission electron microscopy, selected area diffraction pattern analysis, and energy-dispersive X-ray spectroscopic analysis showed two distinct types of prokaryotic cells that precipitated only a U(IV) mineral uraninite (UO2) or both uraninite and metal sulfides. Prokaryotic cells associated with uraninite and metal sulfides were inferred to be sulfate-reducing bacteria. Phylogenetic analysis of 16S ribosomal DNA obtained from the original and incubated sediments revealed that microbial populations were changed from microaerophilic Proteobacteria to anaerobic low-G+C gram-positive sporeforming bacteria by the incubation. Forty-two out of 94 clones from the incubated sediment were related to sulfate-reducing Desulfosporosinus spp., and 23 were related to fermentative Clostridium spp. The results suggest that, if in situ bioremediation were attempted in the uranium mine ponds, Desulfosporosinus spp. would be a major contributor to U(VI) and sulfate reduction and Clostridium spp. to U(VI) reduction.

* Corresponding author. Present address: Frontier Research System for Extremophiles, Japan Marine Science & Technology Center, 2-15, Natushima-cho, Yokosuka 237-006, Japan. Phone: 81(468)67 9710. Fax: 81(468)67 9715. E-mail: yohey{at}jamstec.go.jp.

Applied and Environmental Microbiology, March 2003, p. 1337-1346, Vol. 69, No. 3
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.3.1337-1346.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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