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Applied and Environmental Microbiology, August 2008, p. 5211-5219, Vol. 74, No. 16
0099-2240/08/$08.00+0     doi:10.1128/AEM.00649-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Quantitative Microbial Community Analysis of Three Different Sulfidic Mine Tailing Dumps Generating Acid Mine Drainage{triangledown}

Dagmar Kock and Axel Schippers*

Federal Institute for Geosciences and Natural Resources (BGR), Section Geomicrobiology, Stilleweg 2, 30655 Hannover, Germany

Received 17 March 2008/ Accepted 5 June 2008

The microbial communities of three different sulfidic and acidic mine waste tailing dumps located in Botswana, Germany, and Sweden were quantitatively analyzed using quantitative real-time PCR (Q-PCR), fluorescence in situ hybridization (FISH), catalyzed reporter deposition-FISH (CARD-FISH), Sybr green II direct counting, and the most probable number (MPN) cultivation technique. Depth profiles of cell numbers showed that the compositions of the microbial communities are greatly different at the three sites and also strongly varied between zones of oxidized and unoxidized tailings. Maximum cell numbers of up to 109 cells g–1 dry weight were determined in the pyrite or pyrrhotite oxidation zones, whereas cell numbers in unoxidized tailings were significantly lower. Bacteria dominated over Archaea and Eukarya at all tailing sites. The acidophilic Fe(II)- and/or sulfur-oxidizing Acidithiobacillus spp. dominated over the acidophilic Fe(II)-oxidizing Leptospirillum spp. among the Bacteria at two sites. The two genera were equally abundant at the third site. The acidophilic Fe(II)- and sulfur-oxidizing Sulfobacillus spp. were generally less abundant. The acidophilic Fe(III)-reducing Acidiphilium spp. could be found at only one site. The neutrophilic Fe(III)-reducing Geobacteraceae as well as the dsrA gene of sulfate reducers were quantifiable at all three sites. FISH analysis provided reliable data only for tailing zones with high microbial activity, whereas CARD-FISH, Q-PCR, Sybr green II staining, and MPN were suitable methods for a quantitative microbial community analysis of tailings in general.

* Corresponding author. Mailing address: Federal Institute for Geosciences and Natural Resources (BGR), Stilleweg 2, 30655 Hannover, Germany. Phone: 49 511 643 3103. Fax: 49 511 643 2304. E-mail: a.schippers{at}bgr.de

{triangledown} Published ahead of print on 27 June 2008.

Applied and Environmental Microbiology, August 2008, p. 5211-5219, Vol. 74, No. 16
0099-2240/08/$08.00+0     doi:10.1128/AEM.00649-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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