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Applied and Environmental Microbiology, September 2001, p. 4242-4248, Vol. 67, No. 9
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.9.4242-4248.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Phylogenetic Diversity Analysis of Subterranean Hot Springs in Iceland

Viggó Thór Marteinsson,1,* Sigurbjörg Hauksdóttir,1 Cédric F. V. Hobel,1 Hrefna Kristmannsdóttir,2 Gudmundur Oli Hreggvidsson,1,3 and Jakob K. Kristjánsson1,3

Prokaria Ltd., IS-112 Reykjavík,1 and Orkustofnun2 and Institute of Biology, University of Iceland,3 IS-108 Reykjavík, Iceland

Received 15 February 2001/Accepted 27 June 2001

Geothermal energy has been harnessed and used for domestic heating in Iceland. In wells that are typically drilled to a depth of 1,500 to 2,000 m, the temperature of the source water is 50 to 130°C. The bottoms of the boreholes can therefore be regarded as subterranean hot springs and provide a unique opportunity to study the subterranean biosphere. Large volumes of geothermal fluid from five wells and a mixture of geothermal water from 50 geothermal wells (hot tap water) were sampled and concentrated through a 0.2-µm-pore-size filter. Cells were observed in wells RG-39 (91.4°C) and MG-18 (71.8°C) and in hot tap water (76°C), but no cells were detected in wells SN-4, SN-5 (95 to 117°C), and RV-5 (130°C). Archaea and Bacteria were detected by whole-cell fluorescent in situ hybridization. DNAs were extracted from the biomass, and small-subunit rRNA genes (16S rDNAs) were amplified by PCR using primers specific for the Archaea and Bacteria domains. The PCR products were cloned and sequenced. The sequence analysis showed 11 new operational taxonomic units (OTUs) out of 14, 3 of which were affiliated with known surface OTUs. Samples from RG-39 and hot tap water were inoculated into enrichment media and incubated at 65 and 85°C. Growth was observed only in media based on geothermal water. 16S rDNA analysis showed enrichments dominated with Desulfurococcales relatives. Two strains belonging to Desulfurococcus mobilis and to the Thermus/Deinococcus group were isolated from borehole RG-39. The results indicate that subsurface volcanic zones are an environment that provides a rich subsurface for novel thermophiles.

* Corresponding author. Mailing address: Prokaria Ltd., Gylfaflöt 5, 112 Reykjavík, Iceland. Phone: (354) 5707900. Fax: (354) 5707901. E-mail: viggo{at}prokaria.com.

Applied and Environmental Microbiology, September 2001, p. 4242-4248, Vol. 67, No. 9
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.9.4242-4248.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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