A novel halophilic archaeon, strain MH1-52-1T, was isolated from solar salt imported from Australia. Cells were pleomorphic, non-motile and Gram-negative. Strain MH1-52-1T required at least 3.0 M NaCl and 1 mM Mg2+ for growth. Strain MH1-52-1T was able to grow at pH 4.0–6.0 (optimum, pH 4.4–4.5) and 15–45 °C (optimum, 37 °C). The diether phospholipids phosphatidylglycerol and phosphatidylglycerol phosphate methyl ester, derived from both C20C20 and C20C25 archaeol, were present. Four unidentified glycolipids were also detected. The 16S rRNA gene sequence showed the highest similarity to that of Halobacterium noricense A1T (91.7 %); there were lower levels of similarity to other members of the family Halobacteriaceae. The G+C content of its DNA was 61.4 mol%. Based on our phenotypic, genotypic and phylogenetic analyses, it is proposed that the isolate should be classified as a representative of a new genus and species, for which the name Halarchaeum acidiphilum gen. nov., sp. nov. is proposed. The type strain of Halarchaeum acidiphilum is MH1-52-1T (=JCM 16109T =DSM 22442T =CECT 7534T).

A novel extremely halophilic archaeon, strain 194-10 T , was isolated from a solar salt sample imported into Japan from the Philippines. Strain 194-10 T was pleomorphic, neutrophilic and mesophilic and required at least 10 % (w/v) NaCl but no MgSO 4 . 7H 2 O for growth; it exhibited optimal growth at 15 % (w/v) NaCl and 60 mM MgSO 4 . 7H 2 O. Strain 194-10 T grew at 20-45 6C (optimum, 30 6C) and pH 6.0-9.0 (optimum, pH 6.5-7.0). The G+C content of its DNA was 59.8 mol%. 16S rRNA gene sequence analysis revealed closest proximity to Halostagnicola larsenii XH-48 T (98.5 % similarity), the sole representative of the genus Halostagnicola. Polar lipid analysis revealed that strain 194-10 T contained phosphatidylglycerol and phosphatidylglycerol phosphate methyl ester (the latter derived from both C 20 C 20 and C 20 C 25 archaeol) and several unidentified glycolipids. The results of DNA-DNA hybridization (20.7 % relatedness between Hst. larsenii JCM 13463 T and strain 194-10 T ) and physiological and biochemical characteristics allowed differentiation of strain 194-10 T from Hst. larsenii XH-48 T . Therefore, strain 194-10 T represents a novel species of the genus Halostagnicola, for which the name Halostagnicola kamekurae sp. nov. is proposed, with the type strain 194-10 T (5DSM 22427 T 5JCM 16110 T 5CECT 7536 T ).The haloarchaea constitute a large group of extremely halophilic, aerobic archaea that are placed in the order Halobacteriales, family Halobacteriaceae (Grant et al., 2001). At the time of writing (December 2009), the family Halobacteriaceae contains 27 genera comprising 104 species (Savage et al., 2008;Oren et al., 2009; http://www.the-icsp. org/taxa/halobacterlist.htm; http://www.bacterio.cict.fr/) that display a wide variety of physiological characteristics, including ranges of salinity, temperature and pH for growth. The genus Halostagnicola, belonging to the family Halobacteriaceae, currently contains only one species and one strain, Halostagnicola larsenii XH-48 T , isolated from sediment of Lake Xilinhot, a saline lake in Inner Mongolia, China (Castillo et al., 2006). The strain is pleomorphic, neutrophilic and requires at least 15 % (w/v) NaCl. The highest 16S rRNA gene sequence similarities are obtained with Natrialba aegyptiaca 40 T and Natrialba asiatica 172P1 T . Polar lipids are phosphatidylglycerol and phosphatidylglycerol phosphate methyl ester, the latter derived from both C 20 C 20 and C 20 C 25 archaeol, and two unidentified glycolipids. We isolated a strain closely related to Hst. larsenii XH-48 T from an imported solar salt sample. In this report, we characterize strain 194-10 T and describe its identification. Hst. larsenii XH-48 T was obtained from the Japan Collection of Microorganisms as JCM 13463 T and used as a reference strain.Strain 194-10 T was isolated from a sample of solar salt imported into Japan from the Philippines. The solar salt sample has been on the market from a Japanese firm as 'Mangrove' (http://www.77ochiai.com/page/syouhin06-783gift.htm) as a 200 g package. To 1 g...

A Gram-negative, pleomorphic, aerobic, haloalkaliphilic archaeon, strain T , was isolated from commercial rock salt imported into Japan from China. The genus Halostagnicola belonging to the family Halobacteriaceae was first described by Castillo et al. (2006) and, at the time of writing, the genus comprised two species, Halostagnicola larsenii (Castillo et al., 2006) and Halostagnicola kamekurae (Nagaoka et al., 2010). Cells of the type strains of both species are pleomorphic, neutrophilic and strictly aerobic. In this report, we describe an alkaliphilic haloarchaeon, strain 167-74 T , isolated from commercial rock salt, that belongs to the genus Halostagnicola. Strain 167-74T was isolated from a sample of rock salt produced in Hubei Province, China. The rock salt is labelled 'BEST ancient rock salt' and is sold in Japan by Hanamasa Co., Ltd. The salt sample (1.0 g) was dissolved in 5 ml sterile 5 % NaCl solution and spread on JCM no. 167 medium agar plates. JCM no. 167 medium contains (l O (adjusted to pH 3.6 with HCl). All components, except Na 2 CO 3 , were dissolved in distilled water and made to 900 ml. The medium was adjusted to pH 6.5 with 40 % KOH, and autoclaved. Sodium carbonate was dissolved in 100 ml distilled water and autoclaved separately. After autoclaving, the two solutions were mixed aseptically. The final pH was 9.0. After incubation of agar plates at 37 u C for 2-4 weeks, various coloured colonies developed. Four colonies were transferred to fresh agar plates, and pure cultures were obtained by plating serial dilutions and repeated transfers on agar plates. Partial 16S rRNA gene sequences of the four isolates were almost the same, and the most alkaliphilic strain, 167-74 T , was chosen as a representative for further experiments.Hst. larsenii JCM 13463 T , obtained from the Japan Collection of Microorganisms, and Hst. kamekurae 194-10 T , isolated in our previous study (Nagaoka et al., 2010), were used as reference strains.Abbreviations: PG, phosphatidylglycerol; PGP-Me, phosphatidylglycerol phosphate methyl ester.

Three thermo-tolerant halophilic archaeal strains, SR-441T, SR-412 and SR-188, were isolated from commercial salt samples. Cells were non-motile pleomorphic rod-shaped, and stained Gram-negative. Colonies were pink-pigmented. The three strains were able to grow with 1.7-4.6 M NaCl (optimum, 2.5 M), at pH 6.5-9.0 (optimum, pH 8.0) and at 35-60 °C (optimum, 45 °C). The orthologous 16S rRNA gene sequence similarities amongst the three strains were 98.8-99.3 %, and the level of DNA-DNA relatedness was 71-74 and 72-75 % (reciprocally). The closest relative was Halopiger aswanensis JCM 11628T with 98.6 %-99.1 % similarity in the orthologous 16S rRNA gene sequences, followed by two more Halopiger species, Halopiger xanaduensis JCM 14033T (98.5 %-99.1 %) and Halopiger salifodinae JCM 9578T (95.5 %-95.6 %). DNA-DNA relatednesses between the three strains and H. aswanensis JCM 11628T and H. xanaduensis JCM 14033T were 61 and 54 %, respectively. The polar lipids of the three novel strains were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, and bis-sulfated diglycosyl archaeol-1. The most distinctive feature of the three strains was the ability to grow at 60 °C, while the maximum growth temperature of H. aswanensis is 55 °C. Based on phenotypic and phylogenetic analyses, the isolates are considered to represent a novel species of the genus Halopiger, for which the name Halopiger thermotolerans sp. nov. is proposed. The type strain is SR-441T (=JCM 19583T=KCTC 4248T) isolated from solar salt produced in Australia. SR-412 (=JCM 19582) and SR-188 (=JCM 19581) isolated from commercial salt samples are additional strains of the species.

A Gram-staining-negative, pleomorphic, aerobic, halophilic archaeon, designated strain 294-194-5(T), was isolated in Japan from commercial solar salt imported from the Philippines. Colonies of strain 294-194-5(T) were translucent and red. Strain 294-194-5(T) was able to grow at 20-50 °C (optimum, 37-45 °C), with 14-30 % (w/v) NaCl (optimum, 18 %), and at pH 6.5-8.5 (optimum, pH 8.0). MgCl2 was not required for growth. Phylogenetic analysis based on 16S rRNA gene sequence similarities showed that strain 294-194-5(T) was most closely related to Natronoarchaeum mannanilyticum YSM-123(T) (96.8-97.1 % sequence similarities). The major polar lipids of the novel strain were the C20C20 and C20C25 derivatives of phosphatidylglycerol and phosphatidylglycerol phosphate methyl ester and the same glycolipids (disulfated diglycosyl diether and one unidentified glycolipid) as detected in N. mannanilyticum YSM-123(T). The DNA G+C content of strain 294-194-5(T) was 63.0 mol%. The DNA-DNA relatedness values between the novel strain and N. mannanilyticum YSM-123(T)were 46.5 % and 48.5 % (reciprocal). Based on these data, strain 294-194-5(T) represents a novel species of the genus Natronoarchaeum, for which the name Natronoarchaeum philippinense sp. nov. is proposed. The type strain is 294-194-5(T) ( = JCM 16593(T) = CECT 7630(T)).