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Applied and Environmental Microbiology, June 2002, p. 3046-3054, Vol. 68, No. 6
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.6.3046-3054.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Isolation and Metabolic Characteristics of Previously Uncultured Members of the Order Aquificales in a Subsurface Gold Mine

Ken Takai,1* Hisako Hirayama,1 Yuri Sakihama,2 Fumio Inagaki,1 Yu Yamato,3 and Koki Horikoshi1,2

Sunground Animalcule Retrieval (SUGAR) Project, Frontier Research System for Extremophiles, Japan Marine Science & Technology Center, 2-15 Natsushima-cho, Yokosuka 237-0061,1 Department of Applied Chemistry, Faculty of Engineering, Toyo University, 2100 Kujirai, Kawagoe, Saitama 350-0852,2 Hishikari Mine, Sumitomo Metal Mining Co. Ltd., Maeme 3844, Hishikari-cho, Kagoshima Prefecture 895-2701, Japan3

Received 25 October 2001/ Accepted 4 April 2002

Culture-dependent and -independent techniques were combined to characterize the physiological properties and the ecological impacts of culture-resistant phylotypes of thermophiles within the order Aquificales from a subsurface hot aquifer of a Japanese gold mine. Thermophilic bacteria phylogenetically associated with previously uncultured phylotypes of Aquificales were successfully isolated. 16S ribosomal DNA clone analysis of the entire microbial DNA assemblage and fluorescence in situ whole-cell hybridization analysis indicated that the isolates dominated the microbial population in the subsurface aquifer. The isolates were facultatively anaerobic, hydrogen- or sulfur/thiosulfate-oxidizing, thermophilic chemolithoautotrophs utilizing molecular oxygen, nitrate, ferric iron, arsenate, selenate, and selenite as electron acceptors. Their versatile energy-generating systems may reflect the geochemical conditions of their habitat in the geothermally active subsurface gold mine.

* Corresponding author. Mailing address: Subground Animalcule Retrieval (SUGAR) Project, Frontier Research System for Extremophiles, Japan Marine Science & Technology Center (JAMSTEC), 2-15 Natsushima-cho, Yokosuka 237-0061, Japan. Phone: 81-468-67-9677. Fax: 81-468-66-6364. E-mail: kent{at}jamstec.go.jp.

Applied and Environmental Microbiology, June 2002, p. 3046-3054, Vol. 68, No. 6
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.6.3046-3054.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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