- Volume 61, Issue 1, 2011

Two methane-producing archaea, designated Mic5c12T and Mic6c05T, were isolated from sludge deposited in a crude oil storage tank and a tubercle on the interior of a pipe transporting natural gas-containing brine, respectively. The isolates were Gram-staining-variable, non-motile rods and grew only on H2/CO2. Strain Mic6c05T produced methane from some alcohols without showing any growth; strain Mic5c12T did not utilize alcohols. The optimum growth conditions for strain Mic5c12T were 35 °C, pH 6.5 and 0–0.68 M NaCl and for strain Mic6c05T were 40 °C, pH 6.0–7.5 and 0.34 M NaCl. Strain Mic5c12T was halotolerant and strain Mic6c05T was halophilic. Comparative 16S rRNA gene sequence analysis revealed that strains Mic5c12T and Mic6c05T belonged to the genus Methanobacterium and their closest relative was Methanobacterium subterraneum A8pT (97.3 and 97.9 % 16S rRNA gene sequence similarity, respectively). The findings from the 16S rRNA gene sequence analyses were supported by analysis of McrA, the alpha subunit of methyl-coenzyme M reductase. On the basis of phylogenetic analyses and phenotypic characteristics, two novel species are proposed, Methanobacterium petrolearium sp. nov. and Methanobacterium ferruginis sp. nov., with type strains Mic5c12T (=NBRC 105198T =DSM 22353T) and Mic6c05T (=NBRC 105197T =DSM 21974T), respectively.

A mesophilic, non-motile, hydrogenotrophic, rod-shaped methanogen, designated M2T, was isolated from Holocene permafrost sediments of the Kolyma lowland in the Russian Arctic. Cells were 3–6 μm long and 0.45–0.5 μm wide. Strain M2T grew on H2/CO2 and formate. Optimum conditions for growth were 37 °C, pH 6.8–7.2 and 0.1 M NaCl. The DNA G+C content was 38.1 mol%. On the basis of 16S rRNA gene sequence comparison with known methanogens, strain M2T was affiliated with the genus Methanobacterium and was most closely related to Methanobacterium veterum MK4T and Methanobacterium bryantii DSM 863T (both 99 % 16S rRNA gene sequence similarity). However, no significant DNA–DNA relatedness was observed between strain M2T and these type strains. We propose that strain M2T represents a novel species, with the name Methanobacterium arcticum sp. nov., with type strain M2T (=DSM 19844T =VKM B-2371T).

An aerobic, Gram-negative, yellow-pigmented bacterial strain, designated 0533T, was isolated from frozen soil from the China No. 1 glacier. Phylogenetic analysis of the 16S rRNA gene sequence demonstrated that strain 0533T was a member of the genus Flavobacterium and exhibited 97.1–98.7 % 16S rRNA sequence similarity with its nearest phylogenetic neighbours. Strain 0533T exhibited phenotypic and chemotaxonomic characteristics common to the genus Flavobacterium: menaquinone-6 (MK-6) was the predominant quinone and iso-C15 : 0, C17 : 1 ω6c, anteiso-C15 : 0, iso-C15 : 0 3-OH, C15 : 1 ω6c, iso-C16 : 0 3-OH, summed feature 3 (comprising C16 : 1 ω7c and/or C16 : 1 ω6c), iso-C15 : 1 G and iso-C17 : 0 3-OH were the major fatty acids (>5 %). The DNA G+C content was 32.5 mol%. On the basis of phenotypic and genotypic data, a novel species, Flavobacterium sinopsychrotolerans sp. nov., is proposed. The type strain is 0533T (=CGMCC 1.8704T =JCM 16398T).

A novel bacterial strain, designated M2T2B15T, was isolated from fermented bovine products and was characterized by using a polyphasic approach. Colonies were reddish pink and circular with entire margins. Cells were strictly aerobic, Gram-reaction-negative, oxidase- and catalase-positive rods that lacked flagella and were motile by gliding. Flexirubin-type pigments were absent. 16S rRNA gene sequence analysis indicated that strain M2T2B15T was related most closely to Larkinella insperata LMG 22510T (94.4 % similarity) but shared <87 % similarity with other members of the phylum Bacteroidetes. The major cellular fatty acids were C16 : 1 ω5c, iso-C15 : 0 and iso-C17 : 0 3-OH. The polar lipids were phosphatidylethanolamine, phosphatidylserine, two unidentified aminophospholipids and two unidentified polar lipids. Menaquinone 7 (MK-7) was the major respiratory quinone. The G+C content of the DNA of strain M2T2B15T was 52 mol%. The phenotypic, genotypic and phylogenetic data presented clearly indicate that strain M2T2B15T represents a novel species of the genus Larkinella, for which the name Larkinella bovis sp. nov. is proposed. The type strain is M2T2B15T (=KACC 14040T =NBRC 106324T). Emended descriptions of the genus Larkinella and of Larkinella insperata Vancanneyt et al. 2006 are also proposed.

A Gram-stain-negative, non-flagellated, non-gliding, yellow-pigmented and rod-shaped bacterial strain, designated GSW-R14T, was isolated from seawater of Geoje Island in the South Sea, Korea. Strain GSW-R14T grew optimally at 25 °C, at pH 7.0–8.0 and in the presence of 2 % (w/v) NaCl. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain GSW-R14T belonged to the genus Flavobacterium, joining Flavobacterium gelidilacus LMG 21477T by a bootstrap resampling value of 100 %. Strain GSW-R14T exhibited 97.6 % 16S rRNA gene sequence similarity to F. gelidilacus LMG 21477T and similarities of 91.2–95.2 % to other members of the genus Flavobacterium. Strain GSW-R14T contained MK-6 as the predominant menaquinone. The fatty acid profile of strain GSW-R14T was similar to that of F. gelidilacus LMG 21477T. The DNA G+C content of strain GSW-R14T was 31.4 mol% and its DNA–DNA relatedness with F. gelidilacus LMG 21477T was 31 %. Strain GSW-R14T could be distinguished from F. gelidilacus and the other species of the genus Flavobacterium by its phylogenetic and genetic distinctiveness and by several phenotypic properties. On the basis of these data, strain GSW-R14T is considered to represent a novel species of the genus Flavobacterium, for which the name Flavobacterium ponti sp. nov. is proposed; the type strain is GSW-R14T (=KCTC 22802T =CCUG 58402T).

A Gram-staining-negative, pink bacterium, designated strain BUZ 2T, was isolated from coastal mud from the North Sea (Fedderwardersiel, Germany). Cells were rod-shaped and able to form multicellular filaments. Growth after 7 days was observed at 10–40 °C, at pH 6–8 and with 0–0.5 % NaCl. The phylogenetic tree based on 16S rRNA gene sequences indicated that strain BUZ 2T is a member of the family Cytophagaceae, its closest neighbours being Rudanella lutea 5715S-11T, Spirosoma linguale LMG 10896T and Spirosoma panaciterrae Gsoil 1519T (87.8, 86.4 and 86.1 % sequence similarity, respectively). The major fatty acids were summed feature 3 (comprising C16 : 1 ω7c and/or iso-C15 : 0 2-OH), C16 : 1 ω5c and iso-C15 : 0. The predominant respiratory quinone was MK-7 and the major polar lipids were phosphatidylethanolamine and several unidentified aminophospholipids. The DNA G+C content was 56.5 mol%. On the basis of this polyphasic study, we propose that strain BUZ 2T represents a novel genus and species, for which the name Fibrella aestuarina gen. nov., sp. nov. is proposed. The type strain of Fibrella aestuarina is BUZ 2T (=DSM 22563T =CCUG 58136T). An emended description of the genus Rudanella is also proposed.

A taxonomic study was carried out on strain F44-8T, which was isolated from a crude-oil-degrading consortium, enriched from marine sediment of the Beibu Gulf, PR China. The 16S rRNA gene sequence of strain F44-8T showed highest similarities to those of Flavobacterium frigoris LMG 21922T (93.3 %), Flavobacterium terrae R2A1-13T (93.3 %) and Flavobacterium gelidilacus LMG 21477T (93.1 %). Sequence similarities to other members of the genus Flavobacterium were <93.0 %. The dominant fatty acids of strain F44-8T were iso-C15 : 0, summed feature 3 (iso-C15 : 0 2-OH and/or C16 : 1 ω7c), iso-C15 : 1 G and iso-C17 : 0 3-OH. The DNA G+C content of strain F44-8T was 38.6 mol%. These results are consistent with characteristics of members of the genus Flavobacterium. Strain F44-8T could, however, be readily distinguished from all known Flavobacterium species by a number of phenotypic features. Therefore, according to the phenotypic and 16S rRNA gene sequence data, strain F44-8T represents a novel species in the genus Flavobacterium, for which the name Flavobacterium beibuense sp. nov. is proposed (type strain F44-8T =CCTCC AB 209067T =LMG 25233T =MCCC 1A02877T).

Three Gram-positive, catalase-negative, coccus-shaped organisms were isolated from the oral cavities of bears. The isolates were tentatively identified as a streptococcal species based on the results of biochemical tests. Comparative 16S rRNA gene sequencing studies confirmed that the organisms were members of the genus Streptococcus, but they did not correspond to any recognized species of the genus. The nearest phylogenetic relative of the new isolates was Streptococcus ratti ATCC 19645T (98.6 %), however, DNA–DNA hybridization analysis showed that the isolates displayed less than 15 % DNA–DNA relatedness with the type strain of S. ratti. Colonies of the novel strains grown on mitis salivarius agar showed an extracellular polysaccharide-producing colony morphology. Based on phenotypic and phylogenetic evidence, it is proposed that the novel isolates are classified in the genus Streptococcus as Streptococcus ursoris sp. nov. The type strain of S. ursoris is NUM 1615T (=JCM 16316T=DSM 22768T).

A novel strictly anaerobic, moderately halophilic and mesophilic bacterium, designated strain SOL3f37T, was isolated from a hydrocarbon-polluted soil surrounding a deep petroleum environment located in south Tunisia. Cells of strain SOL3f37T stained Gram-positive and were motile, straight and spore-forming. Strain SOL3f37T had a typical Gram-positive-type cell-wall structure, unlike the thick, multilayered cell wall of its closest relative Clostridiisalibacter paucivorans. The major fatty acids were iso-C15 : 0 (41 %), iso-C14 : 0 3-OH and/or iso-C15 : 0 dimethyl acetal (21.6 %), iso-C13 : 0 (4.4 %), anteiso-C15 : 0 (3.9 %) and iso-C15 : 1 (2.8 %). Strain SOL3f37T grew between 20 and 48 °C (optimum 40 °C) and at pH 6.2–8.1 (optimum pH 6.9). Strain SOL3f37T required at least 0.5 g NaCl l−1 and grew in the presence of NaCl concentrations up to 150 g l−1 (optimum 40 g l−1). Yeast extract (2 g l−1) was required for degradation of pyruvate, fumarate, fructose, glucose and mannitol. Also, strain SOL3f37T grew heterotrophically on yeast extract, peptone and bio-Trypticase, but was unable to grow on Casamino acids. Sulfate, thiosulfate, sulfite, elemental sulfur, fumarate, nitrate and nitrite were not reduced. The DNA G+C content was 30.7 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain SOL3f37T was a member of the family Clostridiaceae in the order Clostridiales; strain SOL3f37T was related to members of various genera of the family Clostridiaceae. It exhibited highest 16S rRNA gene sequence similarity (93.4 %) with Clostridiisalibacter paucivorans 37HS60T, 91.8 % with Thermohalobacter berrensis CTT3T and 91.7 % with Caloranaerobacter azorensis MV1087T. On the basis of genotypic, phenotypic and phylogenetic data, it is suggested that strain SOL3f37T represents a novel species in a new genus. The name Sporosalibacterium faouarense gen. nov., sp. nov. is proposed, with SOL3f37T (=DSM 21485T =JCM 15487T) as the type strain of Sporosalibacterium faouarense.

Two novel thermophilic, spore-forming bacterial strains, T-11T and E-112T, were isolated from hot springs in Tengchong and Eryuan counties of Yunnan province in south-west China. The strains were Gram-stain-positive rods, occurring singly or in chains. Growth of strain T-11T was observed between 30 and 75 °C (optimum 50 °C) and at pH 7–11 (optimum pH 8.5), while the temperature range for strain E-112T was 35–70 °C (optimum 55 °C) and the pH range was 7.0–11.0 (optimum pH 8.0). The DNA G+C contents of strains T-11T and E-112T were 41.1 and 42.6 mol%, respectively. On the basis of 16S rRNA gene sequence similarity, the two strains were shown to be related most closely to Anoxybacillus species. The chemotaxonomic characteristics [predominant isoprenoid quinone menaquinone 7 (MK-7); major fatty acids iso-C15 : 0 and iso-C17 : 0] also supported the affiliation of strains T-11T and E-112T to the genus Anoxybacillus. The results of DNA–DNA hybridization and physiological and biochemical tests allowed genotypic and phenotypic differentiation of strains T-11T and E-112T from Anoxybacillus species with validly published names. Strains T-11T and E-112T therefore represent two novel species, for which the names Anoxybacillus tengchongensis sp. nov. (type strain T-11T =CCTCC AB209237T =KCTC 13721T) and Anoxybacillus eryuanensis sp. nov. (type strain E-112T =CCTCC AB209236T =KCTC 13720T) are proposed.

A xylan-degrading bacterium, strain X11-1T, was isolated from soil collected in Nan province, Thailand. The strain was characterized based on its phenotypic and genotypic characteristics. Strain X11-1T was a Gram-stain-positive, facultatively anaerobic, spore-forming, rod-shaped bacterium. It contained meso-diaminopimelic acid in the cell-wall peptidoglycan. The major menaquinone was MK-7, anteiso-C15 : 0 (56.6 %) and C16 : 0 (14.0 %) were the predominant cellular fatty acids and diphosphatidylglycerol, phosphatidylmonomethylethanolamine, phosphatidylethanolamine and phosphatidylglycerol were the major phospholipids. The DNA G+C content was 51.6 mol%. Phylogenetic analysis using 16S rRNA gene sequences showed that strain X11-1T was affiliated to the genus Paenibacillus and was closely related to Paenibacillus naphthalenovorans KACC 11505T and Paenibacillus validus CCM 3894T, with 96.5 % sequence similarity. Therefore, the strain represents a novel species of the genus Paenibacillus, for which the name Paenibacillus xylanisolvens sp. nov. is proposed. The type strain is X11-1T (=KCTC 13042T =PCU 311T =TISTR 1829T).

A Gram-negative, aerobic bacterium, designated R-40509T, was isolated from mucus of the reef builder coral (Mussismilia hispida) located in the São Sebastião Channel, São Paulo, Brazil. The strain was oxidase-positive and catalase-negative, and required Na+ for growth. Its phylogenetic position was in the genus Marinobacterium and the closest related species were Marinobacterium sediminicola, Marinobacterium maritimum and Marinobacterium stanieri; the isolate exhibited 16S rRNA gene sequence similarities of 97.5–98.0 % with the type strains of these species. 16S rRNA gene sequence similarities with other type strains of the genus Marinobacterium were below 96 %. DNA–DNA hybridizations between strain R-40509T and the type strains of the phylogenetically closest species of the genus Marinobacterium revealed less than 70 % DNA–DNA relatedness, supporting the novel species status of the strain. Phenotypic characterization revealed that the strain was able to grow at 15–42 °C and in medium containing up to 9 % NaCl. The isolate could be differentiated from phenotypically related species by several features, including its ability to utilize d-alanine, l-alanine, bromosuccinic acid, β-hydroxybutyric acid and α-ketovaleric acid, but not acetate or l-arabinose. It produced acetoin (Voges–Proskauer), but did not have esterase lipase (C8) or catalase activities. It possessed C18 : 1 ω7c (35 %), summed feature 3 (iso-C15 : 0 2-OH and/or C16 : 1 ω7c; 25 %) and C16 : 0 (22 %) as major cellular fatty acids. The DNA G+C content was 58.5 mol%. The name Marinobacterium coralli sp. nov. is proposed to accommodate this novel isolate; the type strain is R-40509T (=LMG 25435T =CAIM 1449T).

An analysis of 16S rRNA gene sequences from archived clinical reference specimens has identified two novel Neisseria species. For each species, two strains from independent sources were identified. Amongst species with validly published names, the closest species to the newly identified organisms were Neisseria canis, N. dentiae, N. zoodegmatis, N. animaloris and N. weaveri. DNA–DNA hybridization studies demonstrated that the newly identified isolates represent species that are distinct from these nearest neighbours. Analysis of partial 23S rRNA gene sequences for the newly identified strains and their nearest neighbours provided additional support for the species designation. Bayesian analysis of 16S rRNA gene sequences suggested that the newly identified isolates belong to distinct but related species of the genus Neisseria, and are members of a clade that includes N. dentiae, N. bacilliformis and N. canis. The predominant cellular fatty acids [16 : 0, summed feature 3 (16 : 1ω7c and/or iso-15 : 0 2-OH) and 18 : 1ω7c], as well as biochemical and morphological analyses further support the designation of Neisseria wadsworthii sp. nov. (type strain 9715T =DSM 22247T =CIP 109934T) and Neisseria shayeganii sp. nov. (type strain 871T =DSM 22246T =CIP 109933T).

A novel strain of fermenting, aerotolerant, chemo-organoheterotrophic spirochaete designated PT was isolated from a sulfur ‘Thiodendron’ mat in a saline spring at the Staraya Russa resort (Novgorod Region, Russia). Cells of strain PT exhibited a helical shape. The spirochaete required sulfide in the growth medium and was able to oxidize it non-enzymically to elemental sulfur via the interaction of H2O2 with sulfide and deposit it in the periplasmic space. Growth occurred at 4–32 °C (optimum at 28–30 °C), pH 6.0–8.5 (optimum pH 7.0–7.5), and in 0.1–1 M NaCl (optimum 0.35 M). The isolate used several sugars and polysaccharides as carbon or energy sources but did not use peptides, amino acids, organic acids or alcohols. The products of glucose fermentation were formate, acetate, ethanol, pyruvate, CO2 and H2. The genomic DNA G+C content was 41.7 mol%. 16S rRNA gene sequence analysis showed that strain PT fell within a group of species in the genus Spirochaeta, including Spirochaeta litoralis, S. isovalerica and S. cellobiosiphila, with which it shared less then 89 % sequence similarity. On the basis of its morphology, physiology and other phenotypic properties, as well as its phylogenetic position, the new isolate is considered to represent a novel species of the genus Spirochaeta, for which the name Spirochaeta perfilievii sp. nov. is proposed. The type strain is PT (=DSM 19205T =VKM B-2514T).

A novel, hydrogen- and sulfur-oxidizing bacterium, designated strain 6CT, was isolated from a hot spring in Graendalur, south-western Iceland. Cells of this organism were Gram-reaction-negative, rod-shaped and motile. The strain grew aerobically and was capable of chemolithotrophic growth on thiosulfate and hydrogen, heterotrophic growth on pyruvate, oxalate, acetate and on glutamate in the presence of yeast extract and mixotrophic growth on several organic compounds, thiosulfate and/or hydrogen. During growth on thiosulfate, the final product was sulfate, resulting in a drop in pH from 6.8 to 2.7. Heterotrophic growth on pyruvate was observed at pH 4–7 (optimum pH 4) and 35–50 °C (optimum 45 °C). The DNA G+C content was 65.2 mol%. As determined by 16S rRNA gene sequence analysis, strain 6CT represents a distinct species belonging to the class Betaproteobacteria and is most closely related to Thiomonas intermedia DSM 18155T and Thiomonas perometabolis DSM 18570T. DNA–DNA hybridization between strain 6CT and Thiomonas intermedia DSM 18155T and Thiomonas perometabolis DSM 18570T gave relatedness values below 32 %. These results, together with physiological characteristics, showed that strain 6CT represents a novel species within the genus Thiomonas, for which the name Thiomonas islandica sp. nov. is proposed. The type strain is 6CT (=DSM 21436T =JCM 16107T).

Two Gram-negative, strictly aerobic, non-motile bacteria designated strains KMM 3851T and KMM 3842T were respectively isolated from a marine snail specimen (Umbonium costatum) and from surrounding sediments collected simultaneously from the shore of the Sea of Japan. Phylogenetic analysis based on 16S rRNA gene sequences showed that strains KMM 3851T and KMM 3842T were affiliated with the Roseobacter lineage of the class Alphaproteobacteria as a separate phylogenetic line adjacent to the members of the genus Thalassobacter. These novel isolates shared 98.5 % 16S rRNA gene sequence similarity and 15 % DNA–DNA relatedness. The major isoprenoid quinone of both strains was Q-10 and polar lipids consisted of phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol and aminophospholipids. In addition, strain KMM 3851T contained two unknown lipids, whereas strain KMM 3842T contained diphosphatidylglycerol. Fatty acid analysis revealed C18 : 1 ω7c and C16 : 0 as major components and small amounts of C18 : 2. The DNA G+C contents were 60.4 mol% (KMM 3851T) and 58.5 mol% (KMM 3842T). Based on distinctive phenotypic characteristics, DNA–DNA hybridization data and phylogenetic distance, strains KMM 3851T and KMM 3842T should be classified as representatives of two novel species in a new genus, Litoreibacter gen. nov., with the type species Litoreibacter albidus sp. nov. (type strain KMM 3851T =NRIC 0773T =JCM 16493T) and a second species Litoreibacter janthinus sp. nov. (type strain KMM 3842T =NRIC 0772T =JCM 16492T).

A Gram-reaction-negative, chemo-organotrophic, non-motile, non-spore-forming, rod-shaped bacterium (strain Gsoil 080T) was isolated from soil collected in a ginseng field in Pocheon Province, South Korea, and was investigated by using a polyphasic taxonomic approach. Comparative 16S rRNA gene sequence analysis showed that strain Gsoil 080T was related most closely to Inquilinus limosus strains AU0476T and AU1979 (98.9 % similarity to both). Strain Gsoil 080T shared ≤91.3 % 16S rRNA gene sequence similarity with the type strains of other recognized species examined. The genus Inquilinus belongs to the family Rhodospirillaceae in the order Rhodospirillales, class Alphaproteobacteria. The predominant ubiquinone was Q-10 and the major fatty acids were summed feature 7 (C18 : 1 ω9c/ω12t/ω7c) and C19 : 0 cyclo ω8c. The G+C content of the genomic DNA of strain Gsoil 080T was 69.9 mol%. The level of DNA–DNA relatedness between strain Gsoil 080T and I. limosus LMG 20952T was 12 %. The results of genotypic analyses in combination with chemotaxonomic and physiological data demonstrated that strain Gsoil 080T represents a novel species of the genus Inquilinus, for which the name Inquilinus ginsengisoli sp. nov. is proposed. The type strain is Gsoil 080T (=KCTC 12574T =LMG 23638T).

The distinction and monophyletic property of the basidiomycetous yeast species in the Bulleribasidium clade of the order Tremellales was resolved by molecular phylogenetic analysis based on the combined sequences of the 18S rRNA gene, internal transcribed spacer (ITS) region including 5.8S rRNA gene and 26S rRNA gene D1/D2 domain. The addition to the clade of new anamorphic species identified among ballistoconidium-forming yeasts isolated from China confirmed and strengthened the separation of this clade from other clades or lineages in the order Tremellales. A new anamorphic genus, Mingxiaea gen. nov. (type species Mingxiaea variabilis comb. nov.) is therefore proposed to accommodate the anamorphic species in the Bulleribasidium clade. Six new combinations are proposed for the described species of this clade which were formerly assigned to the genus Bullera. Four novel species in the new genus were identified among 16 ballistoconidium-forming yeast strains isolated from plant leaves collected in Hainan province, southern China, by D1/D2 and ITS sequence analyses. The novel species are described as Mingxiaea sanyaensis sp. nov. (type strain SY-3.23T =AS 2. 3623T =CBS 11408T), Mingxiaea hainanensis (type strain WZS-8.13T =AS 2.4161T =CBS 11409T), Mingxiaea foliicola (type strain WZS-8.14T =AS 2.3518T =CBS 11407T) and Mingxiaea wuzhishanensis (type strain WZS-29.8T =AS 2.4163T =CBS 11411T).