The genus Halobacillus, with the type species Halobacillus halophilus, was established and described by Spring and co-workers in 1996 (Claus et al., 1983;Spring et al., 1996). So far, nine species of this genus have been identified (Amoozegar et al., 2003a, b;Yoon et al., 2003Yoon et al., , 2004Yoon et al., , 2005Liu et al., 2005). The increasing number of publications on bio-applications and other aspects of Halobacillus and the large number of 16S rRNA gene sequences deposited in databases for unidentified species reflect the wide distribution of these bacteria and their considerable scientific interest (Burja et al., 1999;Pinar et al., 2001;Yang et al., 2002; Rivadeneyra et al., 2004). In our investigations of the diversity of halophilic bacteria in various habitats over the last few years, we obtained strains IS-Hb4 T and IS-Hb7 T in an endemic population from a carbonate rock collected at a deep-sea methane seep at Kuroshima Knoll, south-western Japan. Living in the special environment of low temperature, high pressure and low oxygen availability, these bacteria possess several features that are different from those of described species that would be considered evidence of interesting adaptation.Samples from which the bacteria were isolated, carbonate rocks authentically formed in a methane seep of the Kuroshima Knoll, located at 642 m depth at 24 u 07.89 N 124 u 11.29 E (Fujikura et al., 2003;Inagaki et al., 2004), were collected with the remotely operated vehicle Dolphin 3K, operated by the Japan Agency for Marine Earth Science and Abbreviation: CZE, capillary zone electrophoresis.