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Halobacillus profundi sp. nov. and Halobacillus kuroshimensis sp. nov., moderately halophilic bacteria isolated from a deep-sea methane cold seep

¹ Graduate School of Biosphere Science, Hiroshima University, Kagamiyama, Higashi-Hiroshima 739-8528, Japan
² Japan Agency for Marine-Earth Science and Technology, Natsushima-cho, Yokosuka 237-0061, Japan
³ Center for Advanced Marine Core Research, Kochi University, Monobe, Nankoku 783-8502, Japan

Correspondence
Takeshi Naganuma
takn{at}hiroshima-u.ac.jp

Two Gram-positive, rod-shaped, moderately halophilic bacteria were isolated from a deep-sea carbonate rock at a methane cold seep in Kuroshima Knoll, Japan. These bacteria, strains IS-Hb4^T and IS-Hb7^T, were spore-forming and non-motile. They were able to grow at temperatures as low as 9 °C and hydrostatic pressures up to 30 MPa. Based on high sequence similarity of their 16S rRNA genes to those of type strains of the genus Halobacillus, from 96.4 % (strain IS-Hb7^T to Halobacillus halophilus NCIMB 9251^T) to 99.4 % (strain IS-Hb4^T to Halobacillus dabanensis D-8^T), the strains were shown to belong to this genus. DNA–DNA relatedness values of 49.5 % and 1.0–33.0 %, respectively, were determined between strains IS-Hb4^T and IS-Hb7^T and between these strains and other Halobacillus type strains. Both strains showed the major menaquinone MK7 and L-orn–D-Asp cell-wall peptidoglycan type. Straight-chain C_{16 : 0}, unsaturated C_{16 : 1}7c alcohol and C_{18 : 1}7c and cyclopropane C_{19 : 0} cyc fatty acids were predominant in both strains. The DNA G+C contents of IS-Hb4^T and IS-Hb7^T were respectively 43.3 and 42.1 mol%. Physiological and biochemical analyses combined with DNA–DNA hybridization results allowed us to place strains IS-Hb4^T (=JCM 14154^T=DSM 18394^T) and IS-Hb7^T (=JCM 14155^T=DSM 18393^T) in the genus Halobacillus as the respective type strains of the novel species Halobacillus profundi sp. nov. and Halobacillus kuroshimensis sp. nov.

The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene sequences of strains IS-Hb4^T and IS-Hb7^T are AB189298 and AB195680.

Genetic relatedness of strains IS-Hb4^T and IS-Hb7^T to related Halobacillus type strains and differential carbon source utilization patterns are available with the online version of this paper.

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