Halosarcina pallida gen. nov., sp. nov., a halophilic archaeon from a low-salt, sulfide-rich spring A novel halophilic archaeon, strain BZ256 T , was isolated from Zodletone Spring, a sulfide-and sulfur-rich spring in south-western Oklahoma, USA. Cells were non-motile, non-flagellated cocci that divided along two axes, resulting in the formation of sarcina-like clusters. Strain BZ256 T grew at salt concentrations ranging from 1.3 to 4.3 M NaCl, with optimum growth at approximately 3.4 M, and required at least 1 mM Mg 2+ for growth. The pH range for growth was 5.0 to at least 8.5, and the temperature range for growth was 25-45 6C. The two diether phospholipids that are typical of members of the order Halobacteriales, namely phosphatidylglycerol and phosphatidylglycerol phosphate methyl ester, were present in strain BZ256 T , as were two glycolipids chromatographically identical to S-DGD-1 and DGD-1. The 16S rRNA gene sequence of strain BZ256 T showed 96.8 % similarity to that of the type strain of Halogeometricum borinquense, the closest recognized species within the order Halobacteriales. The DNA G+C content of strain BZ256 T was 65.4 mol%. Microscopic, physiological, biochemical and phylogenetic comparisons between strain BZ256 T and recognized genera of extremely halophilic archaea suggest that this strain represents a member of a novel genus and species within the family Halobacteriaceae, for which the name Halosarcina pallida gen. nov., sp. nov. is proposed. The type strain of Halosarcina pallida is BZ256 T (5KCTC 4017 T 5JCM 14848 T ).Members of the family Halobacteriaceae, the single family recognized within the order Halobacteriales, have long been identified as the most abundant micro-organisms in hypersaline environments (Oren, 1994). At the time of writing, the family Halobacteriaceae comprised 25 recognized genera and 101 recognized species. Recently, the extent of the family has been undergoing rapid expansion, with the description (as of June 2007) of five new genera and 20 novel species since 2006. This expansion has been due not only to the identification of novel taxa isolated from well-known hypersaline environments, such as the genera Halovivax, Halostagnicola, Haloplanus and Haloquadratum (Castillo et al., 2006a, b;Bardavid et al., 2007;Burns et al., 2007), but also to the recognition that members of the Halobacteriales can grow within saline microniches in non-saline environments at relatively low salt concentrations (Elshahed et al., 2004a;Savage et al., 2007). In addition, recent studies have reported the isolation of novel halobacterial strains (Purdy et al., 2004;Fukushima et al., 2007) or the detection of novel Halobacteriales-affiliated 16S rRNA gene sequences from moderate-to low-salinity systems (Munson et al., 1997;Takai et al., 2001;Walsh et al., 2005). These studies have shown that the order Halobacteriales is more diverse than previously believed. In the present study, we report on the isolation and characterization of strain BZ256 T from a sulfur-and sulfide-rich spring in south-...