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Applied and Environmental Microbiology, April 2003, p. 1936-1943, Vol. 69, No. 4
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.4.1936-1943.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Enumeration and Characterization of Acidophilic Microorganisms Isolated from a Pilot Plant Stirred-Tank Bioleaching Operation

Naoko Okibe,1 Mariekie Gericke,2 Kevin B. Hallberg,1 and D. Barrie Johnson1*

School of Biological Sciences, University of Wales, Bangor LL57 2UW, United Kingdom,1 Mintek, Randburg 2125, South Africa2

Received 13 August 2002/ Accepted 2 January 2003

Microorganisms were enumerated and isolated on selective solid media from a pilot-scale stirred-tank bioleaching operation in which a polymetallic sulfide concentrate was subjected to biologically accelerated oxidation at 45°C. Four distinct prokaryotes were isolated: three bacteria (an Acidithiobacillus caldus-like organism, a thermophilic Leptospirillum sp., and a Sulfobacillus sp.) and one archaeon (a Ferroplasma-like isolate). The relative numbers of these prokaryotes changed in the three reactors sampled, and the Ferroplasma isolate became increasingly dominant as mineral oxidation progressed, eventually accounting for >99% of plate isolates in the third of three in-line reactors. The identities of the isolates were confirmed by analyses of their 16S rRNA genes, and some key physiological traits (e.g., oxidation of iron and/or sulfur and autotrophy or heterotrophy) were examined. More detailed studies were carried out with the Leptospirillum and Ferroplasma isolates. The data presented here represent the first quantitative study of the microorganisms in a metal leaching situation and confirm that mixed cultures of iron- and sulfur-oxidizing prokaryotic acidophiles catalyze the accelerated dissolution of sulfidic minerals in industrial tank bioleaching operations. The results show that indigenous acidophilic microbial populations change as mineral dissolution becomes more extensive.

* Corresponding author. Mailing address: School of Biological Sciences, University of Wales, Bangor LL57 2UW, United Kingdom. Phone: 44 1248 382358. Fax: 44 1248 370731. E-mail d.b.johnson{at}bangor.ac.uk.

Applied and Environmental Microbiology, April 2003, p. 1936-1943, Vol. 69, No. 4
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.4.1936-1943.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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