Bacillus okhensis sp nov., a halotolerant and alkalitolerant bacterium from an Indian saltpan

A strictly aerobic, rod-shaped bacterium (0.6-0.8 x 2-3 mu m), designated strain Kh10-101(T), was isolated from a saltpan (22 degrees 15' N, 69 degrees 1' E) in the vicinity of Port Okha, India. The creamish pigmented colonies of strain Kh10-101(T) were round, flat and translucent with irregular margins and a smooth surface. The strain possessed up to three subpolar flagella, and was motile by a corkscrew motion. The strain grew optimally at 37 degrees C (temperature growth range 25-40 degrees C in a complex glucose-containing medium with 5% NaCl (NaCI growth range 0-10%) at pH 9 (pH growth range pH 7-10), indicating that it was a mesophilic halotolerant alkaliphile. The strain was sensitive to lincomycin, meticillin, cefuroxime and cephalexin, but resistant to gentamicin, tetracycline and cotrimazine. Spores were not detected and cells were heat sensitive. The isolate metabolized a range of carbohydrates and hydrolysed casein, gelatin and starch. Growth was not observed on aromatic compounds, Tween 40 or Tween 80. Nitrate was not reduced and catalase was produced. Electron microscopic examination of thin sections revealed a single thick Gram-positive cell wall. The DNA G + C content was 41 +/- 1 mol%. Phylogenetic analyses of the 16S rRNA gene sequence revealed that strain Kh10-101(T) was a member of the sixth rRNA group of the genus Bacillus, which includes alkalitolerant, alkaliphilic; and halotolerant species. The halotolerant obligate alkaliphile Bacillus krulwichiae is the closest relative of strain Kh10-101(T) (96% similarity) but a number of phenotypic differences suggest that strain Kh10-101(T) (=JCM 13040(T) =ATCC BAA-1137(T)) should be designated the type strain of a new species, for which the name Bacillus okhensis sp. nov. is proposed.