- Volume 60, Issue 5, 2010

A Gram-staining-positive, aerobic, non-motile and rod-shaped bacterium, designated P27T, was isolated from a traditional fermented seafood. The isolate grew optimally with 0–2.0 % (w/v) NaCl and at pH 6–7 and 30 °C. The predominant menaquinones were MK-12 and MK-11. The major cellular fatty acids were anteiso-C17 : 0, anteiso-C15 : 0 and iso-C16 : 0. The major cell-wall sugars were galactose, mannose and rhamnose. The peptidoglycan amino acids of strain P27T were 2,4-diaminobutyric acid, alanine, glutamic acid and glycine. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and an unidentified glycolipid. The genomic DNA G+C content of strain P27T was 69.0 mol%. Based on its 16S rRNA gene sequence, strain P27T showed highest pairwise similarity with Agromyces cerinus subsp. cerinus JCM 9083T (97.0 % similarity). Based on phenotypic, genotypic and phylogenetic studies, strain P27T represents a novel species in the genus Agromyces, for which the name Agromyces atrinae sp. nov. is proposed. The type strain is P27T (=KCTC 19593T =JCM 15913T).

A novel halophilic actinomycete, designated strain YIM 93223T, was isolated from a salt lake in Xinjiang Province, north-west China, and was subjected to a polyphasic taxonomic study. The isolate grew at 25–45 °C, at pH 6–8 and in the presence of 1–15 % (w/v) NaCl; no growth was observed in the absence of NaCl. Strain YIM 93223T contained meso-diaminopimelic acid, glutamic acid and alanine as cell-wall amino acids, and glucose and galactose as major whole-cell-wall sugars. Major fatty acids were iso-C16 : 0, C16 : 0 and C16 : 1 ω7c/iso-C15 : 0 2-OH. MK-8(H4) was the predominant menaquinone. The genomic DNA G+C content was 66.1 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain YIM 93223T belongs to the genus Amycolatopsis. It shared highest 16S rRNA gene sequence similarity with Amycolatopsis palatopharyngis 1BDZT (96.6 %) and Amycolatopsis marina Ms392AT (96.4 %), but lower values (94.5–96.2 %) with the type strains of other recognized species of the genus Amycolatopsis. On the basis of the data from this polyphasic study, strain YIM 93223T is considered to represent a novel species of the genus Amycolatopsis, for which the name Amycolatopsis halophila sp. nov. is proposed. The type strain is YIM 93223T (=DSM 45216T =KCTC 19403T).

A Gram-positively staining, aerobic, non-motile actinomycete, strain GW 12687T, that formed rose-pigmented colonies and branched substrate and aerial mycelia was studied in detail for its taxonomic position. On the basis of 16S rRNA gene sequence similarity studies, strain GW 12687T was grouped into the genus Nonomuraea, being most closely related to Nonomuraea dietziae (97.6 %), Nonomuraea africana (97.1 %), and Nonomuraea kuesteri (97.1 %). The 16S rRNA gene sequence similarity to other species of the genus Nonomuraea was ≤97 %. The chemotaxonomic characterization supported allocation of the strain to the genus Nonomuraea. The major menaquinone was MK-9(H4) with minor amounts of MK-9(H2), MK-9(H6), MK-9(H0) and MK-8(H4). The polar lipid profile contained the major compound diphosphatidylglycerol, moderate amounts of phosphatidylmonomethylethanolamine, phosphatidylethanolamine, hydroxy-phosphatidylethanolamine, hydroxy-phosphatidylmonomethylethanolamine, and an unknown aminophosphoglycolipid. Phosphatidylinositol mannosides and phosphatidylinositol were also present. The major fatty acids were iso- and anteiso- and 10-methyl-branched fatty acids. The results of physiological and biochemical tests allowed genotypic and phenotypic differentiation of strain GW 12687T from closely related species. Thus, GW 12687T represents a novel species of the genus Nonomuraea, for which the name Nonomuraea rosea sp. nov. is proposed, with GW 12687T (=DSM 45177T =CCUG 56107T) as the type strain.

A novel actinomycete, designated strain 5414T-18T, was isolated from an air sample collected from the Taean region, Korea. The strain contained oxidase and grew in the presence of 7 % NaCl. A neighbour-joining tree constructed on the basis of the 16S rRNA gene sequence showed that strain 5414T-18T is a member of the genus Terrabacter, sharing 97.8–98.3 % 16S rRNA gene sequence similarities to type strains of species of the genus Terrabacter (98.3 % sequence similarity with Terrabacter lapilli LR-26T). It contained peptidoglycan containing ll-diaminopimelic acid of A3γ type, with three glycine residues as the interpeptide bridge. Whole-cell sugars were glucose, mannose and ribose. Mycolic acids were absent. The predominant menaquinone was MK-8(H4). The major fatty acids (>7 % of total fatty acids) were iso-C15 : 0, iso-C16 : 0, C17 : 1 ω8c and iso-C14 : 0. The major polar lipids were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol and an unidentified phosphoglycolipid. The DNA G+C content of the type strain was 73 mol%. Strain 5414T-18T exhibited DNA–DNA relatedness levels of 44, 43, 39, 34 and 34 % to the type strains of Terrabacter lapilli, Terrabacter aerolatus, Terrabacter terrae, Terrabacter tumescens and Terracoccus luteus, respectively. These findings suggest that strain 5414T-18T represents a novel species within the genus Terrabacter. The name Terrabacter aeriphilus sp. nov. is proposed for this novel species, with the type strain 5414T-18T (=KACC 20693T=DSM 18563T).

A coryneform strain, 06-1773OT (=WAL 19168T), derived from a groin abscess sample was characterized using phenotypic and molecular taxonomic methods. Comparative analyses revealed more than 3 % divergence of the 16S rRNA gene sequence and about 10 % divergence of the partial rpoB gene sequence from the type strain of Corynebacterium glucuronolyticum. The strain could also be differentiated from C. glucuronolyticum by a set of phenotypic properties. A DNA–DNA relatedness study between strain WAL 19168T and C. glucuronolyticum CCUG 35055T showed a relatedness value of 13.3 % (13.7 % on repeat analysis). The genotypic and phenotypic data show that the strain merits classification within a novel species of Corynebacterium. We propose the name Corynebacterium pyruviciproducens sp. nov. for the novel species. The type strain is 06-1773OT (=WAL 19168T =CCUG 57046T =ATCC BAA-1742T).

A novel endophytic actinomycete, strain GMKU 367T, was isolated from roots of Thai glutinous rice plants (Oryza sativa L. ‘RD6’) collected from Pathum Thani Rice Research Center, Pathum Thani province, Thailand. Strain GMKU 367T formed cylindrical spores on aerial mycelium, but sporangium-like structures and fragmentation of substrate mycelium were not observed. The cell-wall amino acids contained meso-diaminopimelic acid, alanine, glutamic acid and acetylated muramic acid. The whole-cell sugars were arabinose, galactose, mannose, rhamnose and ribose. Major fatty acids were iso-C15 : 0, iso-C16 : 0 and C16 : 0. The diagnostic menaquinone was MK-9(H4). The polar phospholipids were phosphatidylethanolamine and hydroxyphosphatidylethanolamine. The G+C content of the genomic DNA was 71.1 mol%. Phylogenetic analyses based on 16S rRNA gene sequence data indicated that strain GMKU 367T differed from members of the family Pseudonocardiaceae. On the basis of the evidence presented in this polyphasic study, it is proposed that strain GMKU 367T represents a novel species in a new genus in the family Pseudonocardiaceae, with the name Actinophytocola oryzae gen. nov., sp. nov.; the type strain of Actinophytocola oryzae is GMKU 367T (=BCC 31372T =NBRC 105245T).

A Gram-stain-positive, aerobic actinomycete, strain YIM 60513T, was isolated from the stem of Gloriosa superba L. collected from tropical rainforest at Xishuangbanna, Yunnan Province, south-west China. 16S rRNA gene sequence analysis indicated that strain YIM 60513T belonged to the genus Saccharopolyspora and was closely related to Saccharopolyspora gregorii NCIB 12823T (99.1 % similarity) and Saccharopolyspora cebuensis SPE 10-1T (97.3 % similarity). Data for the predominant quinone [MK-9(H4)], major fatty acids (iso-C16 : 0, anteiso-C17 : 0 and C17 : 1 cis9) and G+C content of the genomic DNA (71.6 mol%) were similar to those for members of the genus Saccharopolyspora. The level of DNA–DNA relatedness between strain YIM 60513T and S. gregorii NCIB 12823T was 43 %. The combination of phylogenetic analysis, phenotypic differences, chemotaxonomic characteristics and DNA–DNA hybridization data supported the view that strain YIM 60513T should be distinguished from S. gregorii NCIB 12823T and S. cebuensis SPE 10-1T. Strain YIM 60513T therefore represents a novel species of the genus Saccharopolyspora, for which the name Saccharopolyspora gloriosae sp. nov. is proposed. The type strain is YIM 60513T (=KCTC 19243T =CCTCC AA 207006T).

Two actinomycete strains, 0351M-12T and 0385M-1T, were isolated from sediment samples collected from Yilan County and Dali City in Taiwan, respectively. The two isolates displayed characteristics which were similar to those of the genus Herbidospora. They both produced branched and unfragmented substrate mycelia, and aerial hyphae were not produced on Bennett's, glucose-asparagine, Hickey-Tresner, sucrose-nitrate, yeast extract-malt extract, or inorganic salts-starch agars. Short spore-chains were borne on the tips of sporophores arising from the substrate mycelia on oatmeal and oatmeal-nitrate agars. The spore chains contained 10–20 non-motile, smooth-surfaced, oval spores. Galactose, glucose, mannose and madurose were the whole-cell sugars in both strains and meso-diaminopimelic acid was present in their peptidoglycans. The phospholipid pattern was PIV. The predominant menaquinone was MK-10(H4) in both strains. Mycolic acids were not detected. Major cellular fatty acids were iso-C16 : 0 and 10-methyl C17 : 0. The DNA G+C contents of the two new isolates were 70.6 mol% (0351M-12T) and 70.7 mol% (0385M-1T), respectively. The relatedness between the isolates and Herbidospora cretacea NBRC 15474T was 40.7–48.5 %. The value of DNA–DNA hybridization between strain 0351M-12T and strain 0385M-1T was 53.6–54.9 %. These two isolates could also be distinguished from each other and from H. cretacea NBRC 15474T by differences in several phenotypic characteristics. We therefore propose the names Herbidospora yilanensis sp. nov. and Herbidospora daliensis sp. nov. for these novel bacteria, with strains 0351M-12T (=FIRDI 003T =BCRC 16875T =LMG 24337T) and 0385M-1T (=FIRDI 004T =BCRC 16876T =LMG 24336T) as the type strains, respectively.

Morphological and chemotaxonomic characterization of actinomycete strain MW2-25T, isolated from tropical forest soil in Nakhon Sawan Province, Thailand, clearly demonstrated that this strain belongs to the genus Dactylosporangium. Phylogenetic analysis using 16S rRNA gene sequences also indicated that this strain should be classified in the genus Dactylosporangium and showed that the closest relative was Dactylosporangium aurantiacum IFO 12592T (99.3 % sequence similarity). DNA–DNA hybridization values and some physiological and biochemical properties indicated that this strain could be readily distinguished from its closest phylogenetic relatives. On the basis of these phenotypic and genotypic data, this strain represents a novel species, for which the name Dactylosporangium maewongense sp. nov. is proposed. The type strain is MW2-25T (=BCC 34832T=JCM 15933T).

Three strains, KV-810T, KV-811 and KV-816, were isolated from mangrove soil from a southern island in Japan on media supplemented with ascorbic acid or rutin. These strains contained l-ornithine as the diagnostic diamino acid in the cell-wall peptidoglycan and DMK-9(H4) as the predominant menaquinone. The G+C content of the DNA was 70–72 mol%. These characteristics in combination with 16S rRNA gene sequence analysis revealed that the novel strains belonged to the genus Demequina. The DNA–DNA hybridization values showed that the three new strains belonged to the same species, a novel species of the genus Demequina. Therefore strains KV-810T, KV-811 and KV-816 are proposed as representing a novel species, Demequina salsinemoris sp. nov. The type strain is KV-810T (=DSM 22060T=NBRC 105323T).

Two extremely halophilic archaea, strains RO1-28T and RO1-22, were isolated from a marine solar saltern in Jiangsu, China. Both strains required at least 0.05 M Mg2+ and 1.7 M NaCl for growth. They were able to grow over a pH range of 6.0–8.5 and a temperature range of 25–55 °C, with optimal pH of 7.0 and optimal temperature of 37–40 °C. Based on 16S rRNA gene sequence analysis, strains RO1-28T and RO1-22 were closely related to Haladaptatus paucihalophilus, the single species of the genus Haladaptatus, with similarities of 94.0–95.2 %. The major polar lipids of the two strains were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and three glycolipids chromatographically identical to the glycolipids of Haladaptatus paucihalophilus JCM 13897T. Both strains RO1-28T and RO1-22 had a DNA G+C content of 54.0 mol% (HPLC). The DNA–DNA hybridization value between the two strains was more than 70 % (92 %) and both strains showed low levels of DNA–DNA relatedness (32 % and 33 %) with Haladaptatus paucihalophilus JCM 13897T. It was concluded that strains RO1-28T and RO1-22 represent a novel species of the genus Haladaptatus, for which the name Haladaptatus litoreus sp. nov. is proposed. The type strain is RO1-28T (=CGMCC 1.7737T =JCM 15771T).

Two isolates of non-alkaliphilic, extremely halophilic archaea, with very similar characteristics, were recovered from a marine solar saltern crystallizer. Cells were pleomorphic, motile and Gram-stain-negative and grew on a limited range of carbon sources, with pyruvate being the best substrate. Optimum growth occurred at 18–20 % (w/v) NaCl, pH 6.0–8.5 and 37–40 °C. Both isolates possessed typical archaeal lipids, and their 16S rRNA gene sequences were 99.8 % identical. Phylogenetic tree reconstructions indicated that they were most closely related to the haloalkaliphile Natronomonas pharaonis (97.5 % similarity to the type strain), but the different phenotypic properties and low DNA–DNA hybridization values between Nmn. pharaonis DSM 2160T and the two isolates suggested that they represent a novel species within the genus Natronomonas. The name Natronomonas moolapensis sp. nov. is proposed for these isolates, with the type strain being 8.8.11T (=JCM 14361T =CECT 7526T =DSM 18674T). An emended description of the genus Natronomonas is also provided.

A novel, extremely halophilic archaeon, D43T, was isolated from traditional salt-fermented seafood in Korea. The cells were Gram-negative-staining and motile. The strain grew at 15–50 °C, 10–30 % (w/v) NaCl and pH 6.0–8.0. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain D43T is affiliated with the family Halobacteriaceae in the domain Archaea and had 95.5 % 16S rRNA gene sequence similarity with Haladaptatus paucihalophilus DX253T. The sequence from strain D43T formed a clade with those from Hap. paucihalophilus regardless of which tree-generating algorithm was used. DNA–DNA hybridization experiments showed 25.8 % relatedness between the isolate and Hap. paucihalophilus KCTC 4006T. Major lipids were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and two unidentified glycolipids. The DNA G+C content of the isolate was 56.5 mol%. On the basis of this polyphasic taxonomic study, strain D43T represents a novel species in the genus Haladaptatus, for which the name Haladaptatus cibarius sp. nov. is proposed. The type strain is D43T (=DSM 19505T =JCM 15962T).

Strains 1.15.5T, 2.27.5, 5.24.4 and 6.14.5 were isolated from a solar saltern. They have flattened, rod-shaped cells and are aerobic, extremely halophilic members of the domain Archaea and family Halobacteriaceae. Cells stained Gram-negative and grew optimally in media around neutral pH and containing 20–24 % (w/v) (strains 1.15.5T and 2.27.5) or 22–24 % (w/v) (5.24.4 and 6.14.5) salts. Mg2+ was not required. The DNA G+C contents of these isolates were all close to 58 mol%, and DNA–DNA cross-hybridization showed a mean relatedness of 77 %. Their 16S rRNA gene sequences differed by no more than 1.6 % from each other. Phylogenetic tree reconstructions with other recognized members of the Halobacteriaceae indicated that they formed a distinct clade, with the closest relative being Halorubrum saccharovorum (86.6–87.6 % 16S rRNA gene sequence similarity to the type strain). The only major polar lipid of all four isolates was the sulfated diglycosyl diether lipid S-DGD-1. By phase-contrast microscopy, the long, flattened cells of these strains often displayed a ‘wing-like’ shape. The phenotypic and phylogenetic data support the placement of these isolates into a novel species in a new genus within the Halobacteriaceae, for which we propose the name Halonotius pteroides gen. nov., sp. nov. The type strain of Halonotius pteroides is 1.15.5T (=JCM 14355T =CECT 7525T =DSM 18729T), with the additional reference strains 2.27.5 (=JCM 14356 =DSM 18671), 5.24.4 (=JCM 14357 =DSM 18673) and 6.14.5 (=JCM 14358 =DSM 18692).

A novel, Gram-positive, non-motile, endospore-forming and moderately halophilic bacterium, strain GrbiT, was isolated from a traditional Korean fermented seafood. The organism grew optimally in the presence of 10–15 % NaCl, at 37 °C and pH 8.0. The peptidoglycan of the cell wall consisted of meso-diaminopimelic acid, and the predominant menaquinone was MK-7. The major fatty acids of strain GrbiT were iso-C16 : 0 (36.4 %), anteiso-C15 : 0 (30.3 %) and iso-C14 : 0 (18.2 %). The polar lipids were phosphatidylglycerol, diphosphatidylglycerol and an unidentified glycolipid. The genomic DNA G+C content was 42.5 mol%. Strain GrbiT was most closely related to the type strain Lentibacillus kapialis JCM 12580T, with which it shared 97.5 % 16S rRNA gene sequence similarity. The DNA–DNA hybridization value between strains GrbiT and L. kapialis JCM 12580T was 8 %. Based on phenotypic, genotypic and phylogenetic data, strain GrbiT should be classified as a novel species within the genus Lentibacillus, for which the name Lentibacillus jeotgali sp. nov. is proposed. The type strain is GrbiT (=KCTC 13300T=JCM 15795T).

Three strains of an unidentified Gram-stain-variable, fastidious, catalase-negative, capnophilic, non-spore-forming, coccus-shaped bacterium from human wound specimens were characterized by phenotypic and molecular taxonomic methods. Initially, these strains were anaerobic; with repeated culture, they became aerotolerant. Comparative 16S rRNA gene sequencing studies demonstrated that the unknown strains were genealogically homogeneous and constituted a novel subline within the genus Gemella. The unknown bacterium was readily distinguished from other Gemella species by biochemical tests. On the basis of both phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium from clinical specimens be classified as Gemella asaccharolytica sp. nov. The type strain is WAL 1945JT (=ATCC BAA-1630T =CCUG 57045T).

Five Gram-stain-positive, motile, aerobic strains were isolated from a clean room of the Kennedy Space Center where the Phoenix spacecraft was assembled. All strains are rod-shaped, spore-forming bacteria, whose spores were resistant to UV radiation up to 1000 J m−2. The spores were subterminally positioned and produced an external layer. A polyphasic taxonomic study including traditional biochemical tests, fatty acid analysis, cell-wall typing, lipid analyses, 16S rRNA gene sequencing and DNA–DNA hybridization studies was performed to characterize these novel strains. 16S rRNA gene sequencing and lipid analyses convincingly grouped these novel strains within the genus Bacillus as a cluster separate from already described species. The similarity of 16S rRNA gene sequences among the novel strains was >99 %, but the similarity was only about 97 % with their nearest neighbours Bacillus pocheonensis, Bacillus firmus and Bacillus bataviensis. DNA–DNA hybridization dissociation values were <24 % to the closest related type strains. The novel strains had a G+C content 35.6±0.5 mol% and could liquefy gelatin but did not utilize or produce acids from any of the carbon substrates tested. The major fatty acids were iso-C15 : 0 and anteiso-C15 : 0 and the cell-wall diamino acid was meso-diaminopimelic acid. Based on phylogenetic and phenotypic results, it is concluded that these strains represent a novel species of the genus Bacillus, for which the name Bacillus horneckiae sp. nov. is proposed. The type strain is 1P01SCT (=NRRL B-59162T =MTCC 9535T).