Two mesophilic, hydrogenotrophic methanogens, designated strains SWAN1T and AL-21, were isolated from two contrasting peatlands: a near circumneutral temperate minerotrophic fen in New York State, USA, and an acidic boreal poor fen site in Alaska, USA, respectively. Cells of the two strains were rod-shaped, non-motile, stained Gram-negative and resisted lysis with 0.1 % SDS. Cell size was 0.6×1.5–2.8 µm for strain SWAN1T and 0.45–0.85×1.5–35 µm for strain AL-21. The strains used H2/CO2 but not formate or other substrates for methanogenesis, grew optimally around 32–37 °C, and their growth spanned through a slightly low to neutral pH range (4.7–7.1). Strain AL-21 grew optimally closer to neutrality at pH 6.2, whereas strain SWAN1T showed a lower optimal pH at 5.4–5.7. The two strains were sensitive to NaCl with a maximal tolerance at 160 mM for strain SWAN1T and 50 mM for strain AL-21. Na2S was toxic at very low concentrations (0.01–0.8 mM), resulting in growth inhibition above these values. The DNA G+C content of the genomes was 35.7 mol% for strain SWAN1T and 35.8 mol% for strain AL-21. Phylogenetic analysis of the 16S rRNA gene sequences showed that the strains are members of the genus Methanobacterium . Strain SWAN1T shared 94–97 % similarity with the type strains of recognized species of the genus Methanobacterium , whereas strain AL-21 shared 99 % similarity with Methanobacterium lacus 17A1T. On the basis of phenotypic, genomic and phylogenetic characteristics, strain SWAN1T ( = DSM 25820T = JCM 18151T) is proposed as the type strain of a novel species, Methanobacterium paludis sp. nov., while strain AL-21 is proposed as a second strain of Methanobacterium lacus .
Halophilic archaeal strain YGHS32T was isolated from the Yinggehai marine solar saltern near Shanya city of Hainan Province, China. Cells of the strain were pleomorphic and lysed in distilled water, stained Gram-negative and formed red-pigmented colonies. Strain YGHS32T was able to grow at 20–50 °C (optimum 37 °C), in the presence of 0.9–4.8 M NaCl (optimum 2.1 M NaCl), with 0.005–1.0 M MgCl2 (optimum 0.3 M MgCl2) and at pH 6.0–8.5 (optimum pH 7.5). The minimal NaCl concentration to prevent cell lysis was 5 % (w/v). The major polar lipids of the strain were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and four major glycolipids chromatographically identical to sulfated mannosyl glucosyl diether, mannosyl glucosyl diether, glucosyl mannosyl glucosyl diether and a diglycosyl diether. Strain YGHS32T had two dissimilar 16S rRNA genes and both of them were phylogenetically related to those of Halomicroarcula pellucida JCM 17820T (92.9–96.3 % sequence similarity). The rpoB′ gene sequence similarity between strain YGHS32T and Halomicroarcula pellucida JCM 17820T was 91.3 %. The DNA G+C content of strain YGHS32T was 64.0 mol%. The DNA–DNA hybridization value between strain YGHS32T and Halomicroarcula pellucida JCM 17820T was 45 %. It was concluded that strain YGHS32T ( = CGMCC 1.12129T = JCM 18640T) represents a novel species of the genus Halomicroarcula , for which the name Halomicroarcula limicola sp. nov. is proposed. An emended description of the genus Halomicroarcula is also presented.
Thermococcus nautili, strain 30-1T (formerly reported as Thermococcus nautilus), was isolated from a hydrothermal chimney sample collected from the East Pacific Rise at a depth of 2633 m on the ‘La chainette PP57’ area. Cells were motile, irregular cocci with a polar tuft of flagella (0.8–1.5 µm) and divided by constriction. The micro-organism grew optimally at 87.5 °C (range 55–95 °C), at pH 7 (range pH 4–9) and with 2 % NaCl (range 1–4 %). Doubling time was 64 min in Zillig’s broth medium under optimal conditions. Growth was strictly anaerobic. It grew preferentially in the presence of elemental sulfur or cystine, which are reduced to H2S, on complex organic substrates such as yeast extract, tryptone, peptone, Casamino acids and casein. Slow growth was observed on starch and pyruvate. Strain 30-1T was resistant to chloramphenicol and tetracyclin (at 100 µg ml−1) but sensitive to kanamycin and rifampicin. The G+C content of the genomic DNA was 54 mol%. Strain 30-1T harboured three plasmids named pTN1, pTN2 and pTN3 and produced membrane vesicles that incorporate pTN1 and pTN3. As determined by 16S rRNA gene sequence analysis, strain 30-1T is related most closely to Thermococcus sp. AM4 (99.3 % similarity) and Thermococcus gammatolerans DSM 15229T (99.2 %). DNA–DNA hybridization values (in silico) with these two closest relatives were below the threshold value of 70 % (33 % with Thermococcus sp. AM4 and 32 % with T. gammatolerans DSM 15229T) and confirmed that strain 30-1 represents a novel species. On the basis of the data presented, strain 30-1T is considered to represent a novel species of the genus Thermococcus , for which the name Thermococcus nautili sp. nov. is proposed. The type strain is 30-1T ( = CNCM 4275 = JCM 19601).