- Volume 63, Issue Pt_1, 2013

An aerobic, rod-shaped, Gram-positive, oxidase- and catalase-positive bacterial isolate, strain YLB-01T, was characterized using phenotypic and molecular taxonomic methods. 16S rRNA gene sequence analysis revealed that the isolate belonged to the genus Microbacterium and represented an evolutionary lineage that was distinct from recognized species of the genus Microbacterium . The isolate showed <97 % 16S rRNA gene sequence similarity with respect to the type strains of all of the members of the genus Microbacterium with validly published names. Cell-wall hydrolysate from the isolate contained the amino acids ornithine, alanine, glycine homoserine and glutamic acid, and the cell-wall sugars consisted of ribose, glucose and galactose. The main respiratory quinones were MK-8, 9, 10 and 11. The major cellular fatty acids were anteiso-C15 : 0 (38.5 %), anteiso-C17 : 0 (23.1 %) and C16 : 0 (18.9 %). The polar lipids contained diphosphatidylglycerol, phosphatidylglycerol, an unidentified phospholipid and two unidentified glycolipids. The DNA G+C content of strain YLB-01T was 71 mol%. On the basis of the morphological, physiological and chemotaxonomic data and the results of comparative 16S rRNA gene sequence analysis, this isolate represents a novel species of the genus Microbacterium , for which the name Microbacterium sediminis sp. nov. is proposed. The type strain is YLB-01T ( = DSM 23767T = CCTCC AB2010363T = MCCC 1A06153T).

A Gram-positive, non-motile, non-spore-forming actinobacterium designated strain Sua-BAC020T was isolated from brine from Sua salt pan in Botswana. The strain was alkaliphilic and moderately halophilic, displaying optimal growth at 35–37 °C, pH 9 and 2.5 % (w/v) NaCl. Comparative 16S rRNA gene sequence analysis showed that strain Sua-BAC020T belonged to the genus Nesterenkonia , sharing 96.2–99.0 % sequence similarity with the type strains of recognized species within this genus. DNA–DNA hybridization with the type strains of species that showed the closest phylogenetic affiliation, Nesterenkonia xinjiangensis (16S rRNA gene sequence similarity, 98.9 %), Nesterenkonia aethiopica (99.0 %), Nesterenkonia halophila (97.5 %), Nesterenkonia flava (97.4 %) and Nesterenkonia halobia (97.2 %), gave relatedness values of 10–45 %. The peptidoglycan type of strain Sua-BAC020T was A4α, l-Lys–Gly–d-Asp. Cells of the isolate contained phosphatidylglycerol, diphosphatidylglycerol, phosphatidylinositol and unidentified glycolipids as major polar lipids, MK-8, MK-9 and MK-7 were the predominant menaquinones, and the major fatty acids (>10 %) were anteiso-C15 : 0 and anteiso-C17 : 0. The DNA G+C content of strain Sua-BAC020T was 64.8 mol%. Based on DNA–DNA hybridization, and physiological and biochemical tests, strain Sua-BAC020T is distinct from all recognized Nesterenkonia species, suggesting that this strain represents a novel species, for which the name Nesterenkonia suensis sp. nov. is proposed. The type strain is Sua-BAC020T ( = DSM 22748T = NCCB 100309T).

A Gram-staining-positive, filamentous bacterium, which developed large globose sporangia at the ends of long sporangiophores on aerial mycelium, was isolated from dry soil collected in a deciduous forest in Thailand. The cell-wall peptidoglycan of the novel bacterium, which was designated strain A-T 1846T, contained meso-diaminopimelic acid and the whole-cell sugars comprised rhamnose, ribose, mannose, glucose and galactose. The predominant menaquinone was MK-9(H4). Mycolic acids were not detected. The identified phospholipids were phosphatidylmethylethanolamine, phosphatidylethanolamine, hydroxyphosphatidylethanolamine, phosphatidylinositol, phosphatidylglycerol and diphosphatidylglycerol. The predominant cellular fatty acids were iso-C16 : 0, iso-C16 : 0 2-OH and C16 : 0 10-methyl. The G+C content of the genomic DNA was 72 mol%. The phenotypic and chemotaxonomic characteristics of the novel strain resembled those of established members of the genus Kutzneria . Phylogenetic analyses based on 16S rRNA gene sequences also indicated that this strain belonged to the genus Kutzneria but as a representative of a putative novel species. In hybridization experiments, the DNA–DNA relatedness values recorded between the novel strain and established members of the genus Kutzneria all fell well below 70 %. Based on the phenotypic, chemotaxonomic and genotypic evidence and the results of the DNA–DNA hybridizations, strain A-T 1846T represents a novel species in the genus Kutzneria , for which the name Kutzneria buriramensis sp. nov. is proposed. The type strain is A-T 1846T ( = BCC 29373T  = NBRC 107931T). An emended description of the genus Kutzneria is provided.

Two novel bacterial strains, designated Kc1T and Kc5T, were isolated from soil in Japan. Cells of the novel strains were Gram-reaction-positive, aerobic or facultatively anaerobic, motile rods. Phylogenetic analyses based on 16S rRNA gene sequences indicated that both strains belonged to the genus Cellulomonas . The 16S rRNA gene sequences of strains Kc1T and Kc5T showed closest similarity to that of Cellulomonas terrae DB5T (98.1 % and 98.4 % similarity, respectively), and the 16S rRNA gene similarity between the two novel strains was 97.8 %. In both strains, the major menaquinone was MK-9(H4), the predominant polar lipids were diphosphatidylglycerol and phosphatidylinositol mannosides, and the peptidoglycan contained ornithine and glutamic acid. Cell-wall sugars were identified as rhamnose, galactose and mannose in strain Kc1T and rhamnose and glucose in strain Kc5T. The DNA G+C contents of strains Kc1T and Kc5T were 73.6 mol% and 75.8 mol%, respectively. Based on the chemotaxonomic and physiological data and the results of DNA–DNA hybridizations, the two strains represent two novel species within the genus Cellulomonas , for which the names Cellulomonas soli sp. nov. (type strain Kc1T  = DSM 24484T  = JCM 17535T) and Cellulomonas oligotrophica sp. nov. (type strain Kc5T = DSM 24482T = JCM 17534T) are proposed.

A novel actinomycete strain, PS7-2T, which produced spore chains borne on the tips of short sporophores, was isolated from soil collected from a temperate peat swamp forest in Phu-Sang National Park, Phayao province, Thailand. The isolate contained glutamic acid, glycine, alanine, 3-hydroxy-diaminopimelic acid and meso-diaminopimelic acid in the cell-wall peptidoglycan. The whole-cell sugars were glucose, mannose, rhamnose and xylose, and the major phospholipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and phosphatidylethanolamine. The predominant menaquinones were MK-10(H8) and MK-9(H8) and the predominant cellular fatty acids were iso-C15 : 0, anteiso-C15 : 0 and anteiso-C17 : 0. The G+C content of the genomic DNA was 72.3 mol%. On the basis of 16S rRNA gene sequence analysis, strain PS7-2T clustered with members of the genus Asanoa and appeared most closely related to the type strains of Asanoa hainanensis (99.5 % sequence similarity), Asanoa iriomotensis (99.0 %), Asanoa ishikariensis (98.9 %) and Asanoa ferruginea (98.5 %). DNA–DNA hybridizations and some physiological and biochemical properties indicated that strain PS7-2T could be readily distinguished from its closest phylogenetic relatives. Based on the phenotypic and genotypic evidence and DNA–DNA relatedness values, strain PS7-2T represents a novel species in the genus Asanoa , for which the name Asanoa siamensis sp. nov. is proposed; the type strain is PS7-2T ( = BCC 41921T = NBRC 107932T).

Three Gram-reaction-positive bacteria, isolated from the air in a medical practice (strains WS4601T, WS4602) or a pharmaceutical clean room (strain WS4599), were characterized using a polyphasic approach. Phylogenetic analyses based on 16S rRNA and recA gene sequences of the three novel strains showed that they formed a distinct lineage within the genus Micrococcus , sharing 16S rRNA gene sequence similarities of 96.1–98.0 % with other species of this genus. Chemotaxonomic features also supported the classification of the three novel strains within the genus Micrococcus . The major cellular fatty acids of strain WS4601T were anteiso-C15 : 0 and iso-C15 : 0, the cell-wall peptidoglycan was of type A3α (l-Lys–l-Ala), and the predominant respiratory quinones were MK-7(H2) and MK-8(H2). The polar lipid profile contained diphosphatidylglycerol and phosphatidylglycerol, but no phosphatidylinositol. The G+C content of the genomic DNA was 70.4 mol%. Numerous physiological properties were found that clearly distinguished strains WS4599, WS4601T and WS4602 from established members of the genus Micrococcus . Based on the phenotypic and phylogenetic data, strains WS4599, WS4601T and WS4602 are considered to represent three different strains of a novel species of the genus Micrococcus , for which the name Micrococcus cohnii sp. nov. is proposed. The type strain is WS4601T ( = DSM 23974T = LMG 26183T).

A novel actinobacterial strain, MBRL 353T, was isolated from a sample collected from a limestone quarry at Hundung, Manipur, India. Comparison of 16S rRNA gene sequences of strain MBRL 353T and other members of the genus Rhodococcus showed sequence similarities ranging from 95.5 to 98.2 %, with strain MBRL 353T showing closest sequence similarity to Rhodococcus triatomae IMMIB RIV-085T (98.2 %) and Rhodococcus equi DSM 20307T (97.2 %). DNA–DNA hybridization results, however, revealed that DNA–DNA relatedness values between strain MBRL 353T and R. triatomae DSM 44892T (43.4 %) and R. equi DSM 20307T (33.4 %) were well below the 70 % limit for species identification. Strain MBRL 353T contained meso-diaminopimelic acid as the diagnostic diamino acid and galactose and arabinose in the cell wall. Mycolic acids were present. The major fatty acids were C16 : 0 (45.7 %), C18 : 1ω9c (18.2 %) and 10-methyl C18 : 0 (11.3 %). The only menaquinone detected was MK-8(H2), while the major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannoside and one unknown phospholipid. The G+C content of the genomic DNA was 69.2 mol%. The phenotypic and genotypic data showed that strain MBRL 353T merits recognition as a representative of a novel species of the genus Rhodococcus for which the name Rhodococcus canchipurensis sp. nov. is proposed; the type strain is MBRL 353T ( = KCTC 19851T = JCM 17578T).

A Gram-positive, non-spore-forming actinobacterium (01-Gi-040T) isolated from an indoor wall was studied to examine its taxonomic position. The isolate formed a very rudimentary substrate-mycelium that fragmented into rod-shaped to coccoid cells. On the basis of the 16S rRNA gene sequence similarity studies, strain 01-Gi-040T was shown to belong to the genus Ornithinimicrobium closely related to Ornithinimicrobium kibberense K22-20T (97.1 %), Ornithinimicrobium humiphilum DSM 12362T (96.2 %) and Ornithinimicrobium pekingense LW6T (96.1 %). A close relationship was also found with Arsenicicoccus bolidensis CCUG 47306T (95.9 %) and Arsenicicoccus piscis Kis4-19T (95.7 %) and a moderate relationship to the type strains of the genus Serinicoccus (94.0–94.1 %). The predominant menaquinone of strain 01-Gi-040T was MK-8(H4). The peptidoglycan contained ornithine as the diagnostic diamino acid. The polar lipid profile consisted of the lipids phosphatidylinositol, phosphatidylglycerol, diphosphatidylglycerol, an unknown phospholipid, an unknown aminolipid and two unknown phosphoglycolipids. The major fatty acids iso-C15 : 0, iso-C16 : 0 and iso-C17 : 0 were consistent with the fatty acid patterns reported for members of the genus Ornithinimicrobium . The results of DNA–DNA hybridizations, physiological and biochemical tests allowed phenotypic differentiation of strain 01-Gi-040T from the three recognized species of the genus Ornithinimicrobium . Strain 01-Gi-040T represents a novel species of the genus Ornithinimicrobium , for which we propose the name Ornithinimicrobium murale sp. nov., with the type strain 01-Gi-040T ( = DSM 22056T = CCM 7610T).

Several intermediate-growing, photochromogenic bacteria were isolated from sphagnum peat bogs in northern Minnesota, USA. Acid-fast staining and 16S rRNA gene sequence analysis placed these environmental isolates in the genus Mycobacterium , and colony morphologies and PCR restriction analysis patterns of the isolates were similar. Partial sequences of hsp65 and dnaJ1 from these isolates showed that Mycobacterium arupense ATCC BAA-1242T was the closest mycobacterial relative, and common biochemical characteristics and antibiotic susceptibilities existed between the isolates and M. arupense ATCC BAA-1242T. However, compared to nonchromogenic M. arupense ATCC BAA-1242T, the environmental isolates were photochromogenic, had a different mycolic acid profile and had reduced cell-surface hydrophobicity in liquid culture. The data reported here support the conclusion that the isolates are representatives of a novel mycobacterial species, for which the name Mycobacterium minnesotense sp. nov. is proposed. The type strain is DL49T ( = DSM 45633T = JCM 17932T = NCCB 100399T).

Two Gram-positive, rod-shaped, non-spore-forming bacteria (strains RHLT1-17T and RHLT2-1T) were isolated from Hailuogou glacier in Szechwan province, PR China. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the strains belonged to the genus Nocardioides and shared 97.8 % sequence similarity with each other and 97.6 and 98.4 % sequence similarity, respectively, with Nocardioides kribbensis KSL-2T. Strain RHLT1-17T grew at 0–35 °C and strain RHLT2-1T grew at 0–25 °C. The major cellular fatty acids of strain RHLT1-17T were C17 : 1ω8c (32.69 %) and iso-C16 : 0 (21.74 %). The major cellular fatty acids of strain RHLT2-1T were C18 : 1ω9c (28.72 %), summed feature 3 (17.14 %; comprising C16 : 1ω7c and/or C16 : 1ω6c), iso-C16 : 0 (14.35 %), C16 : 0 (9.96 %) and iso-C14 : 0 (8.34 %). Both strains contained ll-2,6-diaminopimelic acid in the cell-wall peptidoglycan and MK-8(H4) as the predominant menaquinone. On the basis of data obtained using a polyphasic approach, two novel species, Nocardioides szechwanensis sp. nov. (type strain RHLT1-17T  = CGMCC 1.11147T  = NBRC 108562T) and Nocardioides psychrotolerans sp. nov. (type strain RHLT2-1T  = CGMCC 1.11156T  = NBRC 108563T), are proposed.

A Gram-reaction-positive, non-spore-forming, aerobic actinobacterial strain (2C6-41T) was isolated from the activated sludge from an industrial wastewater treatment plant in Daegu, South Korea. Its taxonomic position was investigated by using a polyphasic approach. On the basis of 16S rRNA gene sequence similarity, closest phylogenetic relatives to strain 2C6-41T were Brevibacterium pityocampae DSM 21720T (97.2 %), Brevibacterium salitolerans KCTC 19616T (96.7 %), Brevibacterium album KCTC 19173T (96.2 %) and Brevibacterium samyangense KCCM 42316T (96.2 %). The DNA G+C content of strain 2C6-41T was 66.4 mol%. Chemotaxonomic data, which included MK-8(H2) as the major menaquinone; meso-diaminopimelic acid, glutamic acid and alanine as cell-wall amino acids; ribose, mannose and glucose as major cell-wall sugars; and anteiso-C15 : 0, anteiso-C17 : 0, C16 : 0 and iso-C15 : 0 as major fatty acids, supported the affiliation of strain 2C6-41T to the genus Brevibacterium . The aromatic ring cleavage enzyme catechol 1,2-dioxygenase was not detected in strain 2C6-41T, but catechol 2,3-dioxygenase was detected. The results of physiological and biochemical tests, and the low level of DNA–DNA relatedness to the closest phylogenetic relative enabled strain 2C6-41T to be differentiated genotypically and phenotypically from recognized species of the genus Brevibacterium . The isolate is therefore considered to represent a novel species in the genus Brevibacterium , for which the name Brevibacterium daeguense sp. nov. is proposed. The type strain is 2C6-41T ( = KCTC 19800T = JCM 17458T).

A slow-growing non-chromogenic mycobacterium was isolated from a patient with pulmonary disease. Phenotypically, strain 05-1390T was similar to Mycobacterium intracellulare ATCC 13950T. The 16S rRNA gene sequence (1385 bp) of strain 05-1390T showed a high degree of similarity to those of the M. intracellulare complex, namely Mycobacterium marseillense 5351974T (100 %), M. intracellulare ATCC 13950T (99.8 %) and Mycobacterium chimaera DSM 44623T (99.9 %). Phylogenetic analysis based on internal transcribed spacer 1 (ITS1) and the hsp65 gene indicated that strain 05-1390T was closely related to M. intracellulare ATCC 13950T, but that it was a distinct phylogenetic entity. Of particular interest, an analysis based on the rpoB gene (701 bp) showed that it is closely related to Mycobacterium parascrofulaceum ATCC BAA-614T (99.4 %), a scotochromogenic strain, rather than to the M. intracellulare -related strains. Unique MALDI-TOF MS profiles also supported the taxonomic status of this strain as a distinct species. These data support the conclusion that strain 05-1390T represents a novel mycobacterial species, for which the name Mycobacterium yongonense sp. nov. is proposed; the type strain is 05-1390T ( = DSM 45126T = KCTC 19555T).

A novel actinomycete, strain A4036T, was isolated from a soil sample collected from the Jabi district in Abuja, Nigeria. The taxonomic position of strain A4036T was established using a combination of genotypic and phenotypic analyses. The organism formed extensively branched substrate and aerial hyphae that generated spiral chains of spores with warty surfaces. The cell wall contained meso-diaminopimelic acid and the cell-wall sugars were glucose, madurose, mannose and ribose. The predominant menaquinone was MK-9(H4). The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylmethylethanolamine, phosphatidylinositol mannoside, hydroxy-phosphatidylethanolamine, hydroxy-phosphatidylmethylethanolamine, two unidentified phospholipids and four unknown glucosamine-containing phospholipids. The major cellular fatty acids were iso-C16 : 0 2-OH, iso-C16 : 0 and 10-methyl C17 : 0. On the basis of 16S rRNA gene sequence similarity studies, strain A4036T grouped in the genus Nonomuraea , being most closely related to Nonomuraea angiospora IFO 13155T (99.05 %), Nonomuraea candida HMC10T (98.78 %), Nonomuraea kuesteri GW 14-1925T (98.49 %), Nonomuraea endophytica YIM 65601T (98.42 %), Nonomuraea maheshkhaliensis 16-5-14T (98.40 %), Nonomuraea turkmeniaca DSM 43926T (98.38 %), Nonomuraea helvata IFO 14681T (98.29 %), Nonomuraea rubra DSM 43768T (98.10 %) and Nonomuraea salmonea DSM 43678T (98.06 %). Levels of 16S rRNA gene sequence similarity to the type strains of other species of the genus Nonomuraea were <98 %. Despite the high 16S rRNA gene sequence similarities, DNA–DNA relatedness values and phenotypic data demonstrated that strain A4036T was clearly distinguished from all closely related species of the genus Nonomuraea . Thus, this isolate is considered to represent a novel species of the genus Nonomuraea , for which the name Nonomuraea jabiensis sp. nov. is proposed. The type strain is A4036T ( = DSM 45507T = KCTC 19870T).

16S rRNA gene sequences deposited for the type strains of Paraoerskovia marina (CTT-37T; GenBank accession no. AB445007) and Koreibacter algae (DSW-2T; FM995611) show a similarity of 100 %. Consequently, the type strains were subjected to a polyphasic recharacterization under direct comparison in order to clarify their taxonomic position. PvuII RiboPrint patterns and quantitative ratios of cellular fatty acids revealed strain-specific differences between P. marina DSM 21750T ( = CTT-37T) and K. algae DSM 22126T ( = DSW-2T). The percentage of DNA–DNA binding of 94 % indicated that the two type strains belong to the same genomospecies. Agreement in the peptidoglycan structure and polar lipid pattern, highly similar fatty acid profiles and MALDI-TOF mass spectra, the ability to produce acid from the same carbon sources, corresponding enzymic activities and DNA G+C contents of 70.8±0.3 mol%, in addition to the consistent characteristics reported in the original descriptions, support the view that the two strains should be affiliated to the same species. According to Rules 38 and 42 of the Bacteriological Code, Koreibacter algae should be reclassified as later heterotypic synonym of Paraoerskovia marina , and the descriptions of the genus Paraoerskovia Khan et al. 2009 and of Paraoerskovia marina Khan et al. 2009 are emended accordingly.

Three filamentous actinomycetes, strains YIM 65594T, YIM 65638 and YIM 65642, were isolated from the surface-sterilized roots of Artemisia annua L. collected from Yunnan province, south-west China. These strains were found to have morphological and chemotaxonomic characteristics typical of the genus Streptomyces . The organisms formed an extensively branched substrate mycelium, with abundant aerial hyphae that differentiated into spores. The cell wall of the isolates contained ll-diaminopimelic acid and the menaquinones were MK-9(H8) and MK-9(H6). The major fatty acids were anteiso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 0. Phylogenetic analysis of the 16S rRNA gene sequences of these strains revealed that the strains clustered together and were most closely related to Streptomyces kunmingensis NBRC 14463T, with 98.5–98.6 % 16S rRNA gene sequence similarity. The results of DNA–DNA hybridization and physiological tests allowed the genotypic and phenotypic differentiation of strains YIM 65594T, YIM 65638 and YIM 65642 from related species. However, the high level of DNA–DNA relatedness between them showed that these three strains belong to the same species. Strain YIM 65594T ( = DSM 41984T = CCTCC AA 209036T) was selected as the type strain to represent this novel species, for which the name Streptomyces endophyticus sp. nov. is proposed.

Two Gram-stain-positive, non-endospore-forming actinobacteria (Ca8Tand Ca14) were isolated from a bioreactor with extensive phosphorus removal. Based on 16S rRNA gene sequence similarity comparisons, strains Ca8T and Ca14 were shown to belong to the genus Gordonia and were most closely related to Gordonia hirsuta DSM 44140T (98.0 % sequence similarity) and Gordonia hydrophobica DSM 44015T (97.2 %). In comparison with the sequences of the type strains of all other species of the genus Gordonia tested, similarities were below 97 %. The quinone systems of the strains were determined to consist predominantly of MK-9H2. The polar lipid profile for both organisms consisted of diphosphatidylglycerol, phosphatidylglycerol, phospatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannoside. Whole-organism hydrolysates contained meso-diaminopimelic acid as the diamino acid of the peptidoglycan; mycolic acids were detected as well. These chemotaxonomic traits and the major fatty acids, which were C16 : 1 cis-9, C16 : 0 and C18 : 1 and tuberculostearic acid strongly supported the grouping of strains Ca8T and Ca14 into the genus Gordonia . The two strains showed a DNA–DNA similarity of 96 %. DNA–DNA hybridizations of strain Ca8T with G. hirsuta DSM 44140T and G. hydrophobica DSM 44015T resulted in values of 26.3 and 25.0 %, respectively. These results and those of the physiological and biochemical tests allowed a clear phenotypic differentiation of strains Ca8T and Ca14 from the most closely related species of the genus Gordonia . It is concluded that strains Ca8T and Ca14 represent a novel species, for which the name Gordonia phosphorivorans sp. nov. is proposed, with the type strain Ca8T ( = DSM 45630T = CCUG 61533T = CCM 7957T = LMG 26648T).