Reassociation Values Research Articles

Mycobacterium oryzae sp. nov., a scotochromogenic, rapidly growing species is able to infect human macrophage cell line.

Gram-stain-positive, acid-fast-positive, rapidly growing, rod-shaped bacteria (designated as strains JC290T, JC430 and JC431) were isolated from paddy cultivated soils on the Western Ghats of India. Phylogenetic analysis placed the three strains among the rapidly growing mycobacteria, being most closely related to Mycobacterium tokaiense 47503T (98.8 % 16S rRNA gene sequence similarity), Mycobacterium murale MA112/96T (98.8 %) and a few other Mycobacterium species. The level of DNA-DNA reassociation of the three strains with M. tokaiense DSM 44635T was 23.4±4 % (26.1±3 %, reciprocal analysis) and 21.4±2 % (22.1±4 %, reciprocal analysis). The three novel strains shared >99.9 % 16S rRNA gene sequence similarity and DNA-DNA reassociation values >85 %. Furthermore, phylogenetic analysis based on concatenated sequences (3071 bp) of four housekeeping genes (16S rRNA, hsp65, rpoB and sodA) revealed that strain JC290T is clearly distinct from all other Mycobacteriumspecies. The three strains had diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositolmannosides, unidentified phospholipids, unidentified glycolipids and an unidentified lipid as polar lipids. The predominant isoprenoid quinone for all three strains was MK-9(H2). Fatty acids were C17 : 1ω7c, C16 : 0, C18 : 1ω9c, C16 : 1ω7c/C16 : 1ω6c and C19 : 1ω7c/C19 : 1ω6c for all the three strains. On the basis of phenotypic, chemotaxonomic and phylogenetic data, it was concluded that strains JC290T, JC430 and JC431 are members of a novel species within the genus Mycobacterium and for which the name Mycobacterium oryzae sp. nov. is proposed. The type strain is JC290T (=KCTC 39560T=LMG 28809T).

Streptococcus halotolerans sp. nov. isolated from the respiratory tract of Marmota himalayana in Qinghai-Tibet Plateau of China.

Two Gramstaining-positive, catalase-negative, α-hemolytic, coccus-shaped organisms were isolated separately from the respiratory tracts of two Marmota himalayana animals from the Qinghai-Tibet Plateau, PR China. Morphological, biological, biochemical, and molecular genetic studies were performed on these two isolates (HTS9T and HTS12). Their biochemical characteristics, such as acid production from different sugars and enzymatic activities, indicated that they represented a member of the genus Streptococcus. They are most closely related to Streptococcus thoraltensis CIP 105518T based on sequence analysis of their 16S rRNA, groEL, sodA and rpoB genes, with similarities of 97.6, 89.9, 92.6 and 91.1 % the four genes respectively. The whole genome phylogenetic tree reconstructed using 372 core genes from 65 genomes of members of the genus Streptococcus validates that HTS9T forms a distinct subline and exhibits specific phylogenetic affinity with S. thoraltensis. In silico DNA-DNA hybridization of HTS9T showed a DNA reassociation value of 32.1 %, closest to that of S. thoraltensis CIP 105518T. Based on their phenotypic characteristics and in particular the phylogenetic findings (DNA-DNA hybridization, three phylogenetic trees built from the partial 16S rRNA/housekeeping genes, and from 372 core genes of 65 genomes of members of the genus Streptococcus), we propose with confidence that strains HTS9T and HTS12 should be classified as representing a novel species of the genus Streptococcus, Streptococcus halotolerans sp. nov. The type strain is HTS9T (=DSM 101996T=CGMCC1.15532T). Genome analysis of Streptococcus halotolerans sp. nov. shows that its genome is 1 823 556 bp long with a DNA G+C content of 39.9 mol% and contains 2068 genes.

Reclassification of Eubacterium desmolans as Butyricicoccus desmolans comb. nov., and description of Butyricicoccus faecihominis sp. nov., a butyrate-producing bacterium from human faeces.

A Gram-positive-staining, coccoid-shaped, non-motile, asporogenous, obligately anaerobic and butyrate-producing bacterium was recovered from a healthy human's faeces. The organism was isolated by the enrichment culture technique using yeast extract-casein hydrolysate-fatty acids broth supplemented with 0.5 % mucin. Phylogenetic analysis of 16S rRNA gene sequences demonstrated that the novel strain should be classified as a member of the Eubacterium desmolans-related cluster in the family Ruminococcaceae. Furthermore, this analysis demonstrated that the type strains of Butyricicoccus pullicaecorum (95.6 %) and Eubacterium desmolans (94.7 %) were the closest phylogenetic neighbours to strain YIT 12789T. However, DNA‒DNA reassociation values with these closest strains were less than 20 %. On the basis of the phenotypic, genotypic and chemotaxonomic features, the novel coccoid-shaped bacterium should be designated as a representative of a novel species of the genus Butyricicoccus, for which the name Butyricicoccus faecihominis sp. nov. is proposed. The type strain is YIT 12789T (=JCM 31056T=DSM 100989T). It is also proposed that Eubacterium desmolans be reclassified in the genus Butyricicoccus as Butyricicoccus desmolans comb. nov.

Paenibacillus silagei sp. nov. isolated from corn silage.

A Gram-stain-negative, rod-shaped, endospore-forming bacterium, strain LOOC204T, was isolated from corn silage that had been prepared and stored in Hokkaido, Japan. The isolate grew well at 4-37 °C and at pH 6.0-8.5. The major cellular fatty acids were anteiso-C15 : 0, C16 : 0, iso-C14 : 0 and iso-C16 : 0. The major menaquinone was MK-7. Diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, three unidentified aminophospholipids, one unidentified phospholipid, two unidentified aminolipids and three unidentified lipids were detected as the cellular polar lipids. The genomic DNA G+C content of LOOC204T was 52.2±0.7 mol%. A comparative 16S rRNA gene sequence analysis showed that LOOC204T represented a member of the genus Paenibacillus, and high levels of sequence similarity of LOOC204T with Paenibacillus borealis KK19T (97.5 %) and Paenibacillusjilunlii Be17T (97.4 %) were found. Based on sequence analysis of the RNA polymerase β subunit (rpoB) gene, LOOC204T was clearly separated from its phylogenetic neighbors in the genus Paenibacillus. DNA-DNA hybridisation experiments showed that LOOC204T displayed less than 40 % reassociation values toward its nearest phylogenetic neighbours of the genus Paenibacillus, indicating the unique taxonomic status of LOOC204T. Based on these data, we conclude that strain LOOC204T represents a novel species of the genus Paenibacillus, named Paenibacillus silageisp. nov. The type strain is LOOC204T (=JCM 30974T=DSM 101953T).

Reclassification of Alteromonas fuliginea (Romanenko et al. 1995) as Pseudoalteromonas fuliginea comb. nov. and an emended description.

A new aerobic marine bacterium, strain S3431, was isolated from swab samples of an unidentified polychaete near Canal Concepción, Chile. This strain was thought to represent a new taxon within the genus Pseudoalteromonas. Although DNA-DNA reassociation values showed less than 70 % genomic DNA relatedness to established Pseudoalteromonas type strains, it shared 78 % DNA-DNA relatedness with Alteromonas fuliginea DSM 15748 (=KMM 216) (Romanenko et al., 1994). A. fuliginea has later been considered a heterotypic synonym of Pseudoalteromonas citrea(Ivanova et al., 1998). Relatedness between strains S3431, A. fuliginea DSM 15748 and the type strain P. citrea LMG 12323T was therefore studied. Physiological traits and genomic information were shared at a high level by strains S3431 and DSM 15748, but not between these and P. citrea LMG 12323T. There was only approximately 20 % DNA-DNA relatedness between P. citrea LMG 12323T and strains S3431 and DSM 15748. Based on the available phylogenetic and phenotypic data, the reclassification of A. fuliginea DSM 15748 (Romanenko et al., 1995) → Pseudoalteromonas citrea(Ivanova et al., 1998) as Pseudoalteromonas fuligineacomb. nov. is proposed, and strain S3431 should be assigned to this new species. The name Pseudoalteromonas fuliginea is proposed with KMM 216T (=DSM 15748T=CIP 105339T) as the type strain.

Marinomonas gallaica sp. nov. and Marinomonas atlantica sp. nov., isolated from reared clams (Ruditapes decussatus).

Three Gram-negative bacterial strains (Cmf 17.2T, Rd 20.33 and Cmf 18.22T) isolated from reared clams in Galicia were subjected to a taxonomic study, based on genetic and phenotypic characterization. Analysis of the 16S rRNA gene allowed the identification of the strains as members of the genus Marinomonas, sharing the highest similarity with Marinomonas aquimarina CECT 5080T (97.8 %-98.5 % 16S rRNA gene sequence similarity). Phylogenetic analysis of the sequences showed that the three isolates formed two different groups distantly related to their closest relative, M. aquimarina. DNA-DNA hybridizations were performed to confirm the taxonomic position and the results were below the recommended threshold for species delimitation, specifically 44.5 % (Cmf 17.2T with M. aquimarina CECT 5080T) and 55 % (Cmf 18.22Twith M. aquimarina CECT 5080T). Furthermore, the average nucleotide identity (ANIb, ANIm and OrthoANI) and in silico estimated DNA-DNA reassociation values among Cmf 17.2T, Cmf 18.22T and M. aquimarina CECT 5080T were in all cases below the respective threshold for species differentiation. The estimated G+C content of the genomic DNA was found to be 45.3 % (Cmf 17.2T) and 44.6 % (Cmf 18.22T). The principal fatty acids of the strains were found to be summed feature 3 (C16 : 1 ω7c/C16 : 1ω6c), summed feature 8 (C18 : 1ω7c/C18 : 1ω6c), C16 : 0, C12 : 0 and C10 : 0 3-OH. The results obtained on the characterization of the clam isolates indicate that they represent two novel species of the genus Marinomonas, for which the names Marinomonas gallaica sp. nov. (type strain Cmf 17.2T=CECT 9049T=LMG 29243T) and Marinomonas atlantica sp. nov. (type strain Cmf 18.22T=CECT 9050T=LMG 29244T) are proposed.

Pelagibacterium lixinzhangensis sp. nov., a Novel Member of the Genus Pelagibacterium.

A Gram-negative, rod shaped and non-spore-forming bacterium, strain H642(T), was isolated from a desert sample collected from Xinjiang province of China, and subjected to a polyphasic taxonomic investigation. Strain H642(T) grew between 20 and 40 °C (optimal 28-37 °C), pH 6.0-10.0 (optimal 7.0-9.0) and at NaCl concentrations between 1.0 and 20.0 % (optimal 2.0-8.0 %). It was positive for catalase, but negative for oxidase. Ubiquinone-10 was present as the major respiratory ubiquinone. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, four unknown glycolipids and one unknown lipid. Phylogenetic analysis of the 16S rRNA gene sequence revealed that strain H642(T) belongs to the genus Pelagibacterium. Strain H642(T) was most closely related to Pelagibacterium halotolerans (with 98.4 % similarity), Pelagibacterium nitratireducens (97.3 %) and Pelagibacterium luteolum (96.2 %). DNA-DNA reassociation values between strain H642(T), P. halotolerans B2(T) and P. nitratireducens JLT2005(T) were 38.0 and 25.3 %, respectively. The genomic DNA G + C content of strain H642(T) was determined to be 44.4 mol%. Based on genotypic, chemotaxonomic and phenotypic data, strain H642(T) represents a novel species of the genus Pelagibacterium, for which the name Pelagibacterium lixinzhangensis sp. nov is proposed. The type strain is H642(T) (=CGMCC 1.10230(T)=JCM 16597(T)).

Leeuwenhoekiella nanhaiensis sp. nov., isolated from deep-sea water.

A novel heterotrophic, aerobic, Gram-stain-negative, rod-shaped and yellow bacterium, designated strain G18T, was isolated from a water sample collected from the deep South China Sea. Strain G18T grew at 4-40 °C (optimum 28-32 °C), at pH 6.0-8.0 (optimum pH 6.5-7.5) and with 0-12 % (w/v) NaCl (optimum 3-4 %). The organism was mesophilic and piezotolerant, its optimal growth pressure was 0.1 MPa, which was lower than that at the depth from which it was isolated. Its optimal growth temperature was higher than that at the depth of its isolation. The predominant cellular fatty acids were C15 : 0iso, C17 : 0iso 3-OH and C15 : 1iso. The major polar lipids were composed of phosphatidylethanolamine, one unknown aminolipid and one unknown polar lipid. The major respiratory quinone was menaquinone 6. The G+C content of the genomic DNA was 35 mol%. Phylogenetic analysis, based on 16S rRNA gene sequences, revealed that strain G18T clustered with species of the genus Leeuwenhoekiella with validly published names within the family Flavobacteriaceae with 95.9-98.2 % sequence similarity. DNA-DNA reassociation values ranged from 9 to 42 %. Differential phenotypic properties, together with the phylogenetic distinctiveness, suggest that strain G18T differs from species of the genus Leeuwenhoekiella with validly published names. On the basis of the polyphasic evidence, strain G18T represents a novel species, isolated from deep-sea, of the genus Leeuwenhoekiella for which the name Leeuwenhoekiella nanhaiensis sp. nov. is proposed. The type strain is G18T ( = CCTCC AB 2015204T = KCTC 42729T).

Bacillus ectoiniformans sp. nov., a halotolerant bacterium isolated from deep-sea sediments.

A halotolerant, Gram-positive bacterium (strain NE-14T), which was isolated from sediment samples of the South China Sea, was subjected to a taxonomic study. Strain NE-14T grew well at wide temperature and pH ranges, 10.0-45.0 °C and pH 6-10, with an optimum at 30 °C and pH 8.0, respectively. Growth of strain NE-14T was observed at total salt concentrations of 0-10 % (w/v) with optimum at 2 % (w/v). Phylogenetic analysis based on 16S rRNA gene sequence comparison indicated that the isolate belongs to the genus Bacillus. Strain NE-14T was related most closely to Bacillus shackletonii LMG 18435T, Bacillus bataviensis LMG 21833T, Bacillus idriensis SMC 4352-2T and Bacillus drentensis LMG 21831T with 16S rRNA gene sequence similarities of 96.2, 95.9, 95.8 and 95.7 %, respectively. DNA-DNA hybridization between strain NE-14T and B. shackletonii LMG 18435T, B. bataviensis LMG 21833T, B. idriensis SMC 4352-2T and B. drentensis LMG 21831T gave reassociation values of about 27.4, 22.4, 16.4 and 15.9 %, respectively. The DNA G+C content of strain NE-14T was 39.2 mol%. The predominant isoprenoid quinone was menaquinone 7 (MK-7). The cell-wall peptidoglycan contained meso-diaminopimelic acid as the diagnostic diamino acid. The predominant cellular fatty acids of strain NE-14T were iso-C15 : 0 and anteiso-C15 : 0. The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and phospholipid. Based on the phenotypic and phylogenetic characteristics, it is proposed that strain NE-14T be classified as representing a novel species of the genus Bacillus, for which the name Bacillus ectoiniformans sp. nov. is proposed. The type strain is NE-14T ( = DSM 28970T = JCM 30397T).

Multilocus sequence analysis of the genus Kurthia, and a description of Kurthia populi sp. nov.

Four novel bacterial strains belonging to the genus Kurthia were isolated from the surface of a weevil of the family Curculionidae (strain 10y-14T), and from bark samples of hybrid poplar, Populus × euramericana (strains 6-3, 2-5 and 06C10-3-14), in Puyang, Henan Province, China. Phylogenetic analyses of the 16S rRNA gene and multilocus sequence analysis (MLSA) data showed that the four strains form a distinct cluster in the genus Kurthia, indicating that they all belong to a single taxon within the genus. DNA-DNA hybridization levels between strain 10y-4T and Kurthia huakuii LAM0618T and Kurthia massiliensis DSM 24639T were 58.31 and 53.92 %, respectively. This indicates that the four novel strains represent a species distinct from these two closely related species. The DNA G+C content of the novel strains was 42.1-42.6 %. The major fatty acids were iso-C15 : 0 and anteiso-C15 : 0.The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, an unknown phospholipid and unidentified aminophospholipids. The predominant menaquinones were MK-7 (90 %) and MK-6 (10 %). The major cell-wall amino acids were lysine, alanine, glutamic acid and glycine. On the basis of the MLSA and 16S rRNA gene sequence phylogenetic analyses, DNA-DNA reassociation values, DNA base composition, and biochemical and phenotypic characteristics, the four strains are considered to represent a novel species within the genus Kurthia, for which the name Kurthia populi sp. nov. is proposed. The type strain is 10y-14T ( = CFCC 11600T = KCTC 33522T).

Ornithinimicrobium algicola sp. nov., a marine actinobacterium isolated from the green alga of the genus Ulva.

A Gram-staining-positive, non-spore-forming actinobacterium, strain JC311T, isolated from marine green alga of the genus Ulva was studied to examine its taxonomic position. On the basis of the 16S rRNA gene sequence similarity studies, strain JC311T was shown represent a member of the genus Ornithinimicrobium and to be closely related to Ornithinimicrobium pekingense LW6T (98.6 %), Ornithinimicrobium kibberense K22-20T (98.3 %) and Ornithinimicrobium humiphilum HKI 0124T (98.1 %). However, strain JC311T showed less than 22 % DNA reassociation value (based on DNA-DNA hybridization) with O. pekingense JCM14001T, O. kibberense JCM12763T and O. humiphilum KCTC19901T. The predominant menaquinone of strain JC311T was MK-8(H4). The peptidoglycan contained l-ornithine as the diagnostic diamino acid. The polar lipid profile consisted of the lipids diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, glycophospholipid, aminophospholipid, phospholipid and two unidentified lipids. The major fatty acids iso-C16 : 0, iso-C15 : 0, iso-C17 : 1ω9c and iso-C17 : 0 were consistent with the fatty acid patterns reported for members of the genus Ornithinimicrobium. The distinct genomic, morphological, physiological and chemotaxonomic differences from the previously described taxa support the classification of JC311T as a representative of a novel species of the genus Ornithinimicrobium, for which we propose the name Ornithinimicrobium algicola sp. nov., with the type strain JC311T ( = KCTC 39559 T = LMG 28808T).

Bacillus endolithicus sp. nov., isolated from pebbles.

Strain JC267T was isolated from pebbles collected from Pingleshwar beach, Gujarat, India. Cells are Gram-stain-positive, facultatively anaerobic, non-motile rods forming sub-terminal endospores in swollen ellipsoidal to oval sporangia. Strain JC267T contains anteiso-C15 : 0, iso-C15 : 0, iso-C14 : 0, iso-C16 : 0, C16 : 0 and anteiso-C17 : 0 as major (>5 %) cellular fatty acids. Polar lipids include phosphatidylglycerol, phospholipids (PL1-3), glycolipids (GL1-2) and an unidentified lipid. Cell-wall amino acids are composed of diagnostic meso-diaminopimelic acid, dl-alanine and a small amount of d-glutamic acid. The genomic DNA G+C content of strain JC267T is 45.5 mol%. The 16S rRNA gene sequence of strain JC267T showed highest sequence similarities of < 98.41 % with all species of the genus Bacillus when subjected to EzTaxon-e blast analysis. The reassociation values based on DNA-DNA hybridization of strain JC267T with Bacillus halosaccharovorans IBRC-M 10095T and Bacillus niabensis JCM 16399T were 26 ± 1 % and 34 ± 3 %, respectively. Based on taxonomic data obtained using a polyphasic approach, strain JC267T represents a novel species of the genus Bacillus, for which the name Bacillus endolithicus sp. nov. is proposed. The type strain is JC267T ( = IBRC-M 10914T = KCTC 33579T).

Aeromicrobium camelliae sp. nov., isolated from Pu'er tea.

A novel Gram-reaction-positive, aerobic and non-spore-forming rod-shaped bacterial strain, YS17T, was isolated from ripened Pu'er tea. Growth of the strain was observed at 15-50 °C (optimum 30-37 °C) and at pH 5.5-10.5 (optimum 6.0-9.5). Phylogenetic analysis of 16S rRNA gene sequences indicated that the strain represented a member of the genus Aeromicrobium. The strains most closely related to YS17T were Aeromicrobium erythreum DSM 8599T, Aeromicrobium alkaliterrae JCM 13518T and Aeromicrobium ginsengisoli JCM 14732T, with 16S rRNA gene sequence similarities of 96.8, 96.8 and 96.7 %, respectively. DNA-DNA hybridization of YS17T with the type strains of the most closely related species, A. erythreum DSM 8599T, A. alkaliterrae JCM 13518T and A. ginsengisoli JCM 14732T, yielded reassociation values of 10.9, 16.8 and 10.9 %, respectively. The diagnostic diamino acid of the cell wall peptidoglycan was ll-diaminopimelic acid. The predominant menaquinones were menaquinone MK-9(H4) (76 %) and MK-8(H4) (17 %). The major fatty acids were C16 : 0, 10-methyl C18 : 0 and C18 : 1ω9c. The DNA G+C content of YS17T was 66 mol%. YS17T could be differentiated from recognized species of the genus Aeromicrobium on the basis of phenotypic characteristics, chemotaxonomic differences, phylogenetic analysis and DNA-DNA hybridization data. On the basis of evidence from the polyphasic analyses performed as part of this study a novel species, Aeromicrobium camelliae sp. nov., is proposed, with strain YS17T ( = CGMCC 1.12942T = JCM 30952T) as the type strain.

Umezawaea endophytica sp. nov., isolated from tobacco root samples.

Two Gram-staining positive, aerobic and non-motile actinomycete strains, designated YIM 2047X(T) and YIM 2047D, were isolated from tobacco root samples collected from Shiling county, Yunnan province, South-West China. The isolates grew at 15-40 °C, pH 6.0-8.0 and 0-4.5 % NaCl (w/v). The whole-cell hydrolysates contained meso-diaminopimelic acid, galactose, mannose, ribose and rhamnose. The polar lipids detected were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylmethylethanolamine, phosphatidylinositol and phosphatidylinositol mannoside. The major cellar fatty acids identified were iso-C14:0, iso-C15:0, anteiso-C15:0, iso-C16:0, C16:0, C17:1 ω8c and C17:0. MK-9 (H4) was the predominant menaquinone. The genomic DNA G+C contents were 73.4 and 74.2 mol%, respectively. These chemotaxonomic data, together with their morphological properties, were consistent with the assignment of the two strains to the genus Umezawaea. They showed highest similarities to Umezawaea tangerina JCM 10302(T) on phylogenetic analysis of their 16S rRNA gene sequences and were found to form a coherent cluster. However, the DNA-DNA hybridization values of YIM 2047X(T) and YIM 2047D with U. tangerina JCM 10302(T) were 46.5 ± 3.1 and 57.5 ± 1.6 %, respectively; while the reassociation value between themselves was 80.4 ± 2.3 %. The results of physiological and biochemical tests allowed the two new isolates to be differentiated phenotypically from the recognized strain U. tangerina JCM 10302(T). On the basis of results from this polyphasic study, the strains were characterized as a novel species of the genus Umezawaea, for which the name Umezawaea endophytica sp. nov. is proposed. The type strain is YIM 2047X(T) (=KCTC 39538(T) = CPCC 204132(T)).

Epilithonimonas psychrotolerans sp. nov., isolated from alpine permafrost.

A bacterial strain, designated TSBY 57T, was isolated during a study on the phylogenetic diversity of culturable bacteria from alpine permafrost in Tianshan Mountains, China, and was classified by means of a polyphasic taxonomic approach. The novel strain was found to belong to the genus Epilithonimonas and was distinguished from recognized species of this genus. Strain TSBY 57T grew aerobically, at 0-30 °C, with 0-1.5% (w/v) NaCl and at pH 6-8.Cells were Gram-stain-negative, non-motile, non-spore-forming rods. Compared with the reference strains, the novel strain was psychrotolerant. The predominant fatty acids were summed feature 3 (consisting of C16:1ω7c and/or C16:1ω6c), anteiso-C15:0 and iso-C15:0.The sole respiratory quinone was MK-6.Phosphatidylethanolamine was predominant in the polar lipid profile of strain TSBY 57T. These chemotaxonomic traits were in good agreement with the characteristics of the genus Epilithonimonas. On the basis of 16S rRNA gene sequence similarity, strain TSBY 57T was a member of the genus Epilithonimonas and was closely related to Epilithonimonas tenax DSM 16811T (99.0%), Epilithonimonas ginsengisoli DCY78T (98.6%) and Epilithonimonas lactis H1T (98.5%). However, DNA-DNA reassociation values between strain TSBY 57T and E. tenax DSM 16811T, E. ginsengisoli DCY78T and E. lactis H1T were 39.5 ± 2.6, 37.7 ± 1.0 and 37.3 ± 1.1%, respectively. The G+C content of the DNA was 34.4 ± 0.2 mol%. Based on data from this polyphasic taxonomic study, strain TSBY 57T represents a novel species of the genus Epilithonimonas, for which the name Epilithonimonas psychrotolerans sp. nov. is proposed. The type strain is TSBY 57T ( = NRRL B-51307T=CCTCC AB 207182T).

Zhihengliuella somnathii sp. nov., a halotolerant actinobacterium from the rhizosphere of a halophyte Salicornia brachiata.

Two novel, Gram-stain-positive, rod-shaped, halotolerent bacteria, strains JG 03(T) and JG 05 were isolated from the rhizosphere of Salicornia brachiata, an extreme halophyte. Comparative analyses of 16S rRNA gene sequences showed that they were closely related to members of the genus Zhihengliuella, with sequence similarities of 96.9-99.1%. The sequence similarity of strains JG 03(T )and JG 05 with each other was 99.4%. DNA-DNA hybridization of JG 03(T) and JG 05 with other species of the genus Zhihengliuella with validly published names showed reassociation values of 19.8%-53.4% and a value of 91.4% between each other. The peptidoglycan type of both strains was A4α and MK-9 and MK-10 were the predominant menaquinones. The predominant fatty acid in JG 03(T) was anteiso-C15 : 0 and anteiso-C17 : 0. However, iso-C15 : 0, anteiso-C15 : 0 and anteiso-C17 : 0 were the major fatty acids in strain JG 05. The DNA G+C content of strains JG 03(T) and JG 05 was 70.0 and 70.1 mol%, respectively. In nutrient broth medium both strains grew at NaCl concentrations of up to 15% (w/v). On the basis of chemotaxonomic characteristics and phylogenetic analyses, strains JG 03(T) and JG 05 should be affiliated to the genus Zhihengliuella. Strains JG 03(T) and JG 05 represent a novel species of the genus Zhihengliuella for which the name Zhihengliuella somnathii sp. nov. is proposed. The type strain is JG 03(T) ( = DSM 23187(T) = IMCC 253(T)).

Alteromonas gracilis sp. nov., a marine polysaccharide-producing bacterium.

A novel exopolysaccharide-producing bacterium, designated strain 9a2(T), was isolated from Pacific Ocean sediment. The strain was Gram-stain-negative, motile, strictly aerobic, oxidase- and catalase-positive, and required NaCl for growth. Its major isoprenoid quinone was ubiquinone-8 (Q-8), and its cellular fatty acid profile consisted mainly of C16 : 1ω7c, C18 : 1ω9c and C16 : 0. The DNA G+C content was 46.6 mol%. 16S rRNA gene sequence analysis suggested that strain 9a2(T) is a member of the genus Alteromonas . Strain 9a2(T) exhibited closest phylogenetic affinity to Alteromonas macleodii NBRC 102226(T) (99.3% 16S rRNA gene sequence similarity), A. marina SW-47(T) (99.3%), A. litorea TF-22(T) (99.0%), A. australica H17(T) (98.7%), A. simiduii BCRC 17572(T) (98.5%), A. stellipolaris LMG 21861(T) (98.3%) and A. hispanica F-32(T) (98.2%). The DNA-DNA reassociation values between strain 9a2(T) and A. macleodii JCM 20772(T), A. marina JCM 11804(T), A. litorea JCM 12188(T), A. australica CIP 109921(T), A. simiduii JCM 13896(T), A. stellipolaris LMG 21861(T) and A. hispanica LMG 22958(T) were below 70%. Strain 9a2(T) contained phosphatidylethanolamine, phosphatidylglycerol and an unidentified polar lipid. Owing to differences in phenotypic and chemotaxonomic characteristics, phylogenetic analysis based on 16S rRNA gene sequences and DNA-DNA relatedness data, the isolate merits classification as representing a novel species, for which the name Alteromonas gracilis sp. nov. is proposed. The type strain of this species is 9a2(T) ( =JCM 30236(T) =NCIMB 14947(T)).

Planifilum caeni sp. nov., a novel member of thermoactinomycete isolated from sludge compost.

A Gram positive, thermophilic, and filamentous bacterium, designated strain GIESS004(T), was isolated from a sludge compost from Guangzhou, China. Growth occurred at 45-55 °C (optimum 50-53 °C) and pH 6.0-10.0 (optimum pH 7.0-7.5). The organism formed cream-yellow colonies with radial wrinkles. Aerial mycelium was not produced in any of the growth media tested. Single spores were produced along the substrate hyphae. Strain GIESS004(T) contained MK-7 as the predominant menaquinone and iso-C(16:0), iso-C(15:0) and iso-C(17:0) as the major cellular fatty acids. The cell wall contained meso-diaminopimelic acid. The DNA G+C content was 60.7 mol%. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain GIESS004(T) was associated with the genus Planifilum and formed a cluster with its closest relative of Planifilum composti P8(T) (99.5 % similarity). However, the DNA-DNA pairing study showed that the isolate displayed reassociation value of only 57.6 ± 2.3 % with P. composti P8(T). On the basis of the polyphasic taxonomic results, strain GIESS004(T) (= CCTCC AB2013107(T) = MCCC 1K00414(T)) is designated as the type strain of a novel species of the genus Planifilum, for which the name Planifilum caeni sp. nov. is proposed.

Microbacterium hydrothermale sp. nov., an actinobacterium isolated from hydrothermal sediment.

A Gram-stain-positive, aerobic, non-motile, rod-shaped bacterium, strain 0704C9-2(T), was isolated from hydrothermal sediment of the Indian Ocean. The organism grew with 0-5% (w/v) NaCl and at 10-37 °C, with optimal growth occurring with 1% NaCl and at 28-30 °C. Comparative 16S rRNA gene sequence analysis revealed that strain 0704C9-2(T) belonged to the genus Microbacterium. It exhibited highest 16S rRNA gene sequence similarity with Microbacterium testaceum DSM 20166(T) (98.4%). Levels of similarity with the type strains of all other recognized species of the genus Microbacterium were less than 98.0%. DNA-DNA hybridization experiments with strain 0704C9-2(T) and its closest relative, M. testaceum DSM 20166(T), revealed a low reassociation value of 42.9%. The DNA G+C content of strain 0704C9-2(T) was 73.3 mol%. The cell-wall peptidoglycan contained ornithine and the acyl type was glycolyl. The major whole-cell sugars were mannose, galactose, rhamnose and glucose. The major cellular fatty acids were anteiso-C15:0, anteiso-C17:0, iso-C16:0 and iso-C15:0. The predominant menaquinones were MK-11, MK-10 and MK-12. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, two unknown glycolipids and an unknown phospholipid. On the basis of phenotypic, phylogenetic and genotypic data, strain 0704C9-2(T) represents a novel species within the genus Microbacterium, for which the name Microbacterium hydrothermale sp. nov. is proposed. The type strain is 0704C9-2(T) ( = LMG 27542(T) = CGMCC 1.12512(T)).

Comparative polyphasic characterization of Streptococcus phocae strains with different host origin and description of the subspecies Streptococcus phocae subsp. salmonis subsp. nov.

A polyphasic study was undertaken to clarify the taxonomic position of Streptococcus phocae strains isolated from Atlantic salmon (Salmo salar) cage-farmed in Chile. Four salmon and three seal isolates showed minor differences in the SDS-PAGE protein analysis. Thus, a major protein band present in the salmon isolates, of approximately 22.4 kDa, was absent in the pinniped strains, regardless of the growth media employed. In addition, the pinniped strains showed protein bands with molecular masses of 71.5 and 14.2 kDa, when grown on trypticase soy agar supplemented with 1% NaCl, or 25.6 kDa, when grown on Columbia blood agar, not present in the Atlantic salmon strains. A high similarity in the matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS spectra of the strains was observed, although some minor peaks were absent in the fish isolates. Fatty acid methyl esters from isolates with different host origin significantly (P<0.05) differed in the content of C16:0, C17:0, C18:1ω9c, C20:4ω6,9,12,15c and summed features 3, 5 and 8. The salmon isolates formed a separate cluster in the phylogenetic analysis of housekeeping genes, separately or as concatenated sequences. Sequence divergences among salmon and seal strains were in the range of inter-subspecies differentiation for groEL (2.5%), gyrB (1.8%), recN (2.1%), rpoB (1.7%) and sodA (2.0%) genes. DNA-DNA hybridization results confirmed those of sequencing, showing reassociation values between seal and salmon strains close to the borderline of species definition. Differences in growth at low temperatures and in the haemolytic capacities were also observed between both groups of isolates. On the basis of all these results, the salmon isolates represent a novel subspecies of S. phocae, for which the name Streptococcus phocae subsp. salmonis subsp. nov. is proposed. The type strain is C-4T (=CECT 7921T=DSM 24768T). The subspecies Streptococcus phocae subsp. phocae subsp. nov. is automatically created. An emended description of S. phocae is also provided.