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Classification of isolates from locations in Austria and Yellowstone National Park as Geobacillus tepidamans sp. nov.

¹ Center for NanoBiotechnology, University of Applied Life Sciences and Natural Resources, A-1180 Wien, Austria
² Thermal Biology Institute, Montana State University, Bozeman, MT 59717-3142, USA
³ Department of Chemistry, University of Applied Life Sciences and Natural Resources, A-1190 Wien, Austria

Correspondence
Paul Messner
paul.messner{at}boku.ac.at

Two moderately thermophilic, Gram-positive, spore-forming bacteria were isolated from different geographical locations and sources; strain GS5-97^T from a beet sugar factory in Leopoldsdorf, Lower Austria, and strain YNP10 from a geothermally heated soil, Yellowstone National Park, USA. The sequences of their 16S rRNA genes were found to be 99·8 % identical, and DNA–DNA hybridization experiments revealed that strains GS5-97^T and YNP10 share 89·9 mol% similarity to each other, but only 34·3 and 39·2 mol% similarity, respectively, to Geobacillus caldoxylosilyticus DSM 12041^T, which is their closest related type strain. A polyphasic analysis showed that these two isolates were more similar to each other than to other characterized geobacilli. Their DNA G+C content was 43·2 and 42·4 mol%, respectively, and they were identical with respect to many phenotypic features (e.g. T_opt 55 °C; pH_opt 7·0). Both strains clearly displayed best growth when cultured aerobically. They differed slightly in their cellular fatty acid profiles and polar lipid pattern, and genotypically they could also be distinguished based on randomly amplified polymorphic DNA fingerprints and internal transcribed spacer analysis. Freeze-etching experiments revealed oblique surface layer (S-layer) lattices in both strains, and biochemical analyses of the purified S-layer proteins indicated the occurrence of glycosylation. Based on the properties of these organisms relative to those currently documented for the genus Geobacillus and for the various sister genera in the Bacillus radiation, a novel species is proposed, Geobacillus tepidamans sp. nov., with GS5-97^T (=ATCC BAA-942^T=DSM 16325^T) as the type strain. Strain YNP10 has been deposited in the American Type Culture Collection as ATCC BAA-943.

The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of Geobacillus tepidamans strain GS5-97^T is AY563003.

Details of media composition, morphological characteristics, chemical analyses and fatty acid compositions are available as supplementary material in IJSEM Online.

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