Two Gram-negative, rod-shaped, halophilic archaea, designated strains HIS40-3 and HDS3-1, were isolated from anchovy fish sauce (nam-pla) collected from two different locations in Thailand. The two strains were able to grow at 20–60 °C (optimum 37–40 °C), at 1.7–5.1 M NaCl (optimum 2.6–3.4 M NaCl) and at pH 5.5–8.5 (optimum pH 6.0–6.5). Hypotonic treatment with less than 1.7 M NaCl caused cell lysis. The major polar lipids of the isolates were CC and CC derivatives of phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate, two glycolipids and one unidentified lipid. The DNA G+C contents were 64.0–65.4 mol%. In addition to phenotypic and chemotaxonomic characteristics, phylogenetic analysis based on 16S rRNA gene sequence similarities showed that strains HIS40-3 and HDS3-1 were related most closely to species of the genus . Levels of 16S rRNA gene sequence similarity between strains HIS40-3 and HDS3-1 and the type strains of recognized species were 99.1–96.6 %. The two novel strains could be distinguished from recognized species on the basis of low levels of DNA–DNA relatedness and differences in whole-cell protein patterns and phenotypic properties. Levels of 16S rRNA gene sequence similarity and DNA–DNA relatedness between the two strains were 99.7 and 77.7 %, respectively, suggesting that they should be classified as representing a single species. Based on these taxonomic data, strains HIS40-3 and HDS3-1 are considered to represent a novel species of the genus , for which the name sp. nov. is proposed. The type strain is HIS40-3 (=BCC 24370 =JCM 14663 =PCU 303).


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Two-dimensional TLC of polar lipids extracted from strain HIS40-3 (a) and strain HDS3-1 (b). Separation of components was achieved by developing the plate with chloroform/methanol/water (65:25:4 by vol., horizontal direction) and then with chloroform/methanol/acetic acid/water (85:12:15:4 by vol., vertical). Spots were visualized by staining with anisaldehyde reagent and the plate was heated to 150 °C for 10 min to show glycolipids and phospholipids. PG, Phosphatidylglycerol; PGP-Me, phosphatidylglycerol phosphate methyl ester; PGS, phosphatidylglycerol sulfate; GL, unidentified glycolipid; Un, unknown lipid; Pi, pigment.


Phylogenetic trees showing the relationships between strains HIS40-3 and HDS3-1 and related archaeal species based on 16S rRNA gene sequences. [PDF](20 KB)


Whole-cell proteins extracted from strains HIS40-3 and HDS3-1 and strains of related species, following separation by SDS-PAGE (10 %). Proteins were stained with Coomassie blue. Lanes: 1, molecular mass markers; 2, JCM 8980 ; 3, JCM 10476 ; 4, JCM 12890 ; 5, strain HIS40-3 ; 6, strain HDS3-1. Arrows indicated major differences in protein bands.

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