- Volume 63, Issue Pt_12, 2013

A novel bacterial strain, designated M8T, was isolated from milk of a female macaque bred in captivity. The strain was Gram-stain-positive, anaerobic, irregular coccoid–rod-shaped without catalase activity. Analysis of 16S rRNA gene sequence similarity revealed that the isolate was most closely related to Alloscardovia omnicolens CCUG 31649T (96.4 %) and Metascardovia criceti OMB105T (96.6 %). Sequences of hsp60, fusA, and xfp genes also confirmed that the strain was most closely related to the type strains of A. omnicolens and M. criceti . The isolate produced fructose-6-phosphate phosphoketolase which is in agreement with classification within the family Bifidobacteriaceae . The major fatty acids were C18 : 1ω9c (35.8 %), C16 : 1 (6.2 %) and C14 : 0 (5.7 %). Polar lipid analysis revealed five different glycolipids, two unidentified phospholipids and diphosphatidylglycerol. The peptidoglycan was of the type A4α l-Lys–d-Asp with the presence of d(l)-alanine, d-glutamine, d-asparagine and l-lysine. The DNA G+C content of strain M8T was 50.1 mol%. On the basis of genetic, phylogenetic and phenotypic data, strain M8T represents a novel species of the genus Alloscardovia for which the name Alloscardovia macacae sp. nov. is proposed. The type strain is M8T ( = DSM 24762T = CCM 7944T). In addition, our results also revealed that Alloscardovia omnicolens DSM 21503T and Metascardovia criceti DSM 17774T do not belong to different genera within the family Bifidobacteriaceae . We therefore propose to reclassify Metascardovia criceti as Alloscardovia criceti comb. nov. An emended description of the genus Alloscardovia is also provided.

Two actinomycete strains isolated from marine sediment samples, designated YIM M11335T (from the Indian Ocean) and 12A09T (from the South China Sea), were obtained and examined by a polyphasic approach. The two Gram-staining-positive, aerobic strains produced branched substrate mycelia and aerial hyphae that were not fragmented, and no diffusible pigment was produced on the media tested. At maturity, spore chains and single spores were formed on aerial hyphae and substrate mycelium, respectively. Whole-organism hydrolysates of both strains contained meso-diaminopimelic acid and the diagnostic sugars glucose and galactose. Their predominant menaquinones were MK-10(H4), MK-10(H6), MK-11(H4), MK-11(H6) and MK-11(H8) for strain YIM 11335T and MK-10(H4), MK-10(H6), MK-11(H4), MK-11(H6) and MK-11(H8) for strain 12A09T. The polar lipids detected in the two strains were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol mannoside, phosphatidylinositol, phosphatidylcholine, an unknown phosphoglycolipid and several unknown glycolipids, phospholipids and polar lipids. The major fatty acids (>10 %) were iso-C16 : 0 and C16 : 0 for strain YIM 11335T and iso-C16 : 0 for strain 12A09T. The G+C contents of the genomic DNA of strains YIM 11335T and 12A09T were 70.7 % and 74.4 %, respectively. DNA–DNA hybridization relatedness values of these two isolates with the type strains Nocardiopsis arabia DSM 45083T and Streptomonospora halophila YIM 91355T supported the hypothesis they are representatives of two different species. Based on phylogenetic analysis, phenotypic and genotypic data, it is concluded that the two isolates belong to the genus Streptomonospora of the family Nocardiopsaceae and that the type strain of N. arabia should be reclassified as a representative of Streptomonospora arabica comb. nov. The names proposed for the two novel species are Streptomonospora sediminis sp. nov. (type strain YIM M11335T = DSM 45723T = CCTCC AB 2012051T) and Streptomonospora nanhaiensis sp. nov. (type strain 12A09T = KCTC 29145T = CCTCC AB 2013140T), respectively. An emended description of the genus Streptomonospora is also proposed in the light of the new data.

A Gram-positive, strictly aerobic and heterotrophic, non-spore-forming actinobacterium (strain B2T) isolated from a recirculating aquaculture system was studied for its taxonomic position. Strain B2T formed a rudimentary substrate-mycelium that fragmented into short rod-shaped to coccoid cells (0.5 µm×0.5–2.2 µm or 0.5–1.0 µm in diameter). Colonies were yellow, smooth, circular and 1.5–2.0 mm in diameter after incubation on TSA for 3 days at 30 °C. Strain B2T grew at 20–40 °C (optimal, 30 °C) and pH 5.5–9.5 (optimal, 6.5–7.0) and in the presence of 0–9 % (w/v) NaCl (optimal, 1 %). The predominant menaquinone of strain B2T was MK-8(H4). The cell-wall peptidoglycan of strain B2T contained the amino acids ornithine, glutamic acid, alanine, glycine and aspartic acid. The major polar lipids were phosphatidylglycerol and diphosphatidylglycerol. The major fatty acids were iso-C15 : 0, iso-C16 : 0 and summed feature 9. Its DNA G+C content was 68.3 mol% (T m). Analysis of 16S rRNA gene sequences indicated that strain B2T was related phylogenetically to members of the genus Ornithinimicrobium with highest similarity (96.6 %) to Ornithinimicrobium kibberense DSM 17687T, followed by Ornithinimicrobium humiphilum DSM 12362T (96.3 %), Ornithinimicrobium pekingense LW6T (96.2 %) and Ornithinimicrobium murale 01-Gi-040T (94.8 %). On basis of phenotypic, chemotaxonomic and phylogenetic data, it was concluded that strain B2T represents a novel species of the genus Ornithinimicrobium , for which the name Ornithinimicrobium tianjinense sp. nov. is proposed. The type strain is B2T ( = CGMCC 1.12160T = JCM 18464T).

Two groups of Gram-stain positive, aerobic bacterial strains were isolated from raw cow’s milk, from a milking machine and from bulk tank milk. Based on their 16S rRNA gene sequences these isolates formed two distinct groups within the genus Corynebacterium . The sequence similarities of the isolates to the type strains of species of the genus Corynebacterium were below 98.4 %. The presence of menaquinones MK-8(H2) and MK-9(H2), the predominant fatty acid 18 : 1 cis 9 and a polar lipid pattern with several phospholipids but without aminolipids was in accord with the characteristics of this genus. The results of DNA–DNA hybridization, biochemical tests and chemotaxonomic properties allowed genotypic and phenotypic differentiation of the strains from all known species of the genus Corynebacterium . Therefore, the isolates were assigned to two novel species of this genus for which the names Corynebacterium frankenforstense sp. nov. (type strain ST18T = DSM 45800T = CCUG 63371T), and Corynebacterium lactis sp. nov. (type strain RW2-5T = DSM 45799T = CCUG 63372T) are proposed, respectively.

Strain MBRL 34T, isolated from a sample of limestone quarry located at Hundung, Manipur, India, was characterized by polyphasic taxonomy. The strain showed the highest 16S rRNA gene sequence similarity with Micromonospora echinaurantica DSM 43904T (98.4 %), but formed a monophyletic clade with Micromonospora coerulea DSM 43143T (98.3 %) in the neighbour-joining tree. DNA–DNA hybridization experiments gave a DNA–DNA relatedness value of 53.1 % between MBRL 34T and M. coerulea DSM 43143T. Strain MBRL 34T contained meso-diaminopimelic acid, galactose and glucose in the whole-cell hydrolysates along with small amounts of mannose, xylose, rhamnose and ribose. The major polar lipids detected were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol and phosphatidylinositolmannoside, along with an unknown lipid. MK-10(H6), MK-10(H2) MK-11(H4) and MK-10(H4) were the predominant menaquinones detected. The major fatty acids were iso-C16 : 0 and iso-C15 : 0. The G+C content of the genomic DNA was 73.5 %. Based on the taxonomic characteristics from a polyphasic study, strain MBRL 34T merits recognition as a representative of a novel species of the genus Micromonospora for which the name Micromonospora kangleipakensis sp. nov. is proposed; the type strain is MBRL 34T ( = DSM 45612T = JCM 17696T).

Three independent strains of a rapidly growing, non-chromogenic member of the genus Mycobacterium were isolated from lymph nodes of French cattle. Identification of the isolates was carried out using a polyphasic approach. The nearly complete SSU rRNA gene sequences (>1200 bp) of the strains MLB-A23, MLB-A30 and MLB-A84T were identical. A phylogenetic analysis of these unique SSU rRNA gene sequences showed that these strains were most closely related to Mycobacterium intermedium . Further phylogenetic analysis based on concatenated sequences (2854 bp) of four housekeeping genes (hsp65, rpoB, sodA and tuf), the transfer–messenger RNA (tmRNA) and SSU rRNA genes indicated that these three strains represented a distinct species that shares a common ancestor with M. intermedium . Phylogenetic and phenotypic data strongly indicate that the strains MLB-A23, MLB-A30 and MLB-A84T belong to a novel mycobacterial species for which the name Mycobacterium bourgelatii sp. nov. is proposed. The type strain is MLB-A84T ( = CIP 110557T = DSM 45746T).

A novel actinomycete, designated Z4T, was isolated from soil in Yaan, Sichuan Province, south China. The taxonomic status of this strain was established using a polyphasic approach. The morphological and chemotaxonomic characteristics of the organism are typical of the members of the genus Streptomyces . Phylogenetic analysis based on the almost complete 16S rRNA gene sequence indicated that strain Z4T belonged to the genus Streptomyces , branching off next to Streptomyces durhamensis ATCC 23194T (98.7 %), Streptomyces puniciscabiei KACC 20253T (98.7 %) and Streptomyces filipinensis ATCC 23905T (98.6 %). However, DNA–DNA hybridization studies and phenotypic differences between strain Z4T and closely related species of the genus Streptomyces suggested that strain Z4T represented a different genomic species. It is therefore proposed that Z4T ( = CGMCC 4.7035T = KCTC 29111T) represents the type strain of a novel species of the genus Streptomyces , for which the name Streptomyces yaanensis sp. nov. is proposed.

Two Gram-stain-positive, non-motile, non-spore-forming, rod-shaped actinobacterial strains were isolated from the surface-sterilized roots of mugwort (Artemisia princeps) and horse-weed (Conyza canadensis), and subjected to taxonomic characterization. 16S rRNA gene sequence analysis indicated that the isolates, designated MWE 3-5T and HWE 2-02T, should be placed in the genus Nocardioides of the family Nocardioidaceae . The strains were closely related to Nocardioides hankookensis DS-30T, which exhibited 16S rRNA gene sequence similarity values of 97.99 and 99.09 % with strains MWE 3-5T and HWE 2-02T, respectively. The genome relatedness of N. hankookensis DS-30T with strain MWE 3-5T was 35.8 %, and that with strain HWE 2-02T was 36.4 %, whereas that between the two isolates was 43.2 %. Strains MWE 3-5T and HWE 2-02T possessed MK-8(H4) as the major isoprenoid quinone, and ll-diaminopimelic acid in the cell-wall peptidoglycan. The main fatty acids were iso-C16 : 0, iso-C17 : 0 and C18 : 1ω9c for strain MWE 3-5T and iso-C16 : 0, 10-methyl C18 : 0 and C18 : 1ω9c for strain HWE 2-02T. Based on phenotypic, genotypic and phylogenetic studies, the following two novel species are proposed: Nocardioides endophyticus sp. nov. (type strain, MWE 3-5T = KCTC 29122T = JCM 18532T) and Nocardioides conyzicola sp. nov. (type strain, HWE 2-02T = KCTC 29121T = JCM 18531T).

A Gram-stain-positive, aerobic, non-motile, non-spore-forming, short rod-shaped actinobacterium, designated strain PB243T, was isolated from grass soil sampled in Daejeon, Republic of Korea. Comparative 16S rRNA gene sequence studies showed the isolate was clearly affiliated with the class Actinobacteria , and most closely related to Amnibacterium kyonggiense KEMC 51201-037T, showing 98.8 % 16S rRNA gene sequence similarity. Cells of strain PB243T formed yellow colonies on R2A agar, contained MK-11 and MK-12 as the predominant menaquinones, l-2,4-diaminobutyric acid as the diagnostic cell-wall diamino acid, and anteiso-C15 : 0 and iso-C16 : 0 among the major fatty acids. The acyl type of the muramic acid was acetyl. The G+C content of the genomic DNA of strain PB243T was 71.5 mol%. Thus, the combined genotypic and phenotypic data supported the conclusion that strain PB243T represents a novel species of the genus Amnibacterium , for which the name Amnibacterium soli sp. nov. is proposed. The type strain is PB243T ( = KCTC 33147T = JCM 19015T).

Cellulolytic bacteria A191T, A192 and A193 isolated from the soil of Sakhalin fir forest in Hokkaido, Japan were studied phenotypically, genotypically and phylogenetically. Analysis of their 16S rRNA gene and gyrB sequences and DNA base composition suggested that these isolates were conspecific and members of the genus Streptomyces . However, levels of 16S rRNA gene and gyrB sequence similarity between the isolates and the type strains of their closest relatives in the genus Streptomyces were no higher than 97.9 and 95.0 %, respectively, implying that these isolates were distinctive. Moreover, the results of DNA−DNA hybridization experiments and physiological characterization clearly differentiated these isolates from their closest neighbours. It is therefore concluded that these isolates represent a novel species of the genus Streptomyces , for which the name Streptomyces abietis is proposed. The type strain is A191T ( = NBRC 109094T = DSM 42080T).

A novel Gram-staining-positive actinobacterium, designated H85-3T, was isolated from a sea sediment sample and its taxonomic position was investigated by a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain H85-3T was closely related to the members of the genus Zhihengliuella with pairwise sequence similarities of 97.4–98.6 %. The peptidoglycan of strain H85-3T was found to be of the A4α type with lysine as the diagnostic diamino acid. The menaquinones were MK-9, MK-10 and MK-8 (56 : 30 : 14) and the major cellular fatty acids were anteiso-C15 : 0, iso-C15 : 0 and anteiso-C17 : 0. These data supported the affiliation of strain H85-3T to the genus Zhihengliuella . Meanwhile, the results of DNA–DNA hybridization, along with the differences in some phenotypic characteristics, indicated that strain H85-3T should be distinguished from the recognized species of the genus Zhihengliuella . Therefore, strain H85-3T represents a novel species of the genus Zhihengliuella , for which the name Zhihengliuella flava sp. nov. is proposed; the type strain is H85-3T ( = NBRC 109021T = DSM 26152T). An emended description of the genus Zhihengliuella is also proposed.

A facultatively anaerobic, alkaliphilic, Gram-stain-positive, rod-shaped bacterium, designated strain 12/1T, isolated from alkaline wastewater drained sludge of a beverage industry facility located near New Delhi, India, was subjected to a polyphasic taxonomic study. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain 12/1T belonged to the genus Exiguobacterium and was most closely related to Exiguobacterium aurantiacum DSM 6208T (99.46 %), E. aquaticum IMTB-3094T (99.18 %), E. mexicanum 8NT (99.06 %), E. profundum 10CT (98.17 %), E. aestuarii TF-16T (98.1 %) and E. marinum TF-80T (98.03 %). The DNA G+C content of strain 12/1T was 55.6 mol%, major respiratory isoprenoid quinone was MK-7, major polar lipids were phosphatidylglycerol, diphosphatidylglycerol and phosphatidylethanolamine and the cell-wall peptidoglycan was of the A3α l-Lys–Gly type, characteristics consistent with its affiliation to the genus Exiguobacterium . Strain 12/1T showed levels of DNA–DNA hybridization of less than 70 % with the closely related species of the genus Exiguobacterium . Overall, the phenotypic, chemotaxonomic and phylogenetic data presented in this study suggest that strain 12/1T represents a novel species of the genus Exiguobacterium , for which the name Exiguobacterium alkaliphilum sp. nov. is proposed. The type strain is 12/1T ( = CCM 8459T = DSM 21148T).

A bacterial strain, designated PECAE04T, was isolated from root nodules of Cicer arietinum in Spain. Phylogenetic analysis based on 16S rRNA gene sequence placed the isolate into the genus Paenibacillus with its closest relative being Paenibacillus castaneae Ch-32T with 98.4 % 16S rRNA gene sequence similarity followed by Paenibacillus glycanilyticus DS-1T, Paenibacillus prosopidis PW21T, Paenibacillus xinjiangensis B538T and Paenibacillus catalpae D75T with similarities ranging from 97.9 to 96.8 %. DNA–DNA hybridization measurements showed values lower than 20 % between the strain PECAE04T and any of these species. The isolate was a Gram-stain-positive, motile, sporulating rod. Catalase and oxidase activities were positive. Aesculin was hydrolysed but casein and gelatin were not. Acetoin production, H2S production, nitrate reduction and urease and caseinase production were negative. Growth was supported by many carbohydrates and organic acids as carbon sources. MK-7 was the predominant menaquinone and anteiso-C15 : 0, iso-C16 : 0 and C16 : 0 were the major fatty acids. Major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, a glycolipid, three phospholipids and an unidentified lipid. Meso-diaminopimelic acid was not detected in the peptidoglycan. The DNA G+C content was 52.9 mol%. Phylogenetic, chemotaxonomic and phenotypic analyses showed that strain PECAE04T should be considered to be a representative of a novel species of the genus Paenibacillus , for which the name Paenibacillus endophyticus sp. nov. is proposed. The type strain is PECAE04T ( = LMG 27297T = CECT 8234T).

An obligately alkaliphilic, anaerobic, thermo- and halotolerant, spore-forming bacterium was isolated from sediments of soda lake Magadi (Kenya) and designated strain Z-1001T. Cells of strain Z-1001T were straight, Gram-positive rods, slowly motile. Strain Z-1001T was found to be an obligate anaerobe. It grew within a pH range from 7.5 to 10.7 with an optimum at 9.25–9.5 (at 40 °C), a temperature range from 20 to 57 °C with an optimum at 45–50 °C, and a NaCl concentration range from 0 to 1.55 M with an optimum at 1.2–1.4 M. Peptides, such as meat and yeast extracts, peptone and tryptone, were fermented by Z-1001T. Carbohydrates did not support growth. With yeast extract as an electron donor, strain Z-1001T reduced , , , Fe(III) citrate and anthraquinone-2,6-disulfonate (AQDS) as electron acceptors. The isolate was able to grow oligotrophically with a very small amount of yeast extract: 0.03 g l−1. The main fatty acids were C16 : 0, C16 : 1ω7c , C18 : 0 and C18 : 1ω9. The DNA G+C content of the isolate was 35.6 mol%. 16S rRNA gene sequence analysis showed that strain Z-1001T is a member of family Natranaerobiaceae , clustering with the type strain of Natranaerobius thermophilus (95.8–96.0 % sequence similarity). On the basis of physiological and phylogenetic data it is proposed that strain Z-1001T ( = DSM 24923T = VKM B-2666T) represents a novel genus and species, Natranaerobaculum magadiense gen. nov., sp. nov.

Three lactic acid bacteria were isolated from faeces of a jackal (Canis mesomelas) and raccoons (Procyron lotor). The isolates formed a subcluster in the Lactobacillus salivarius phylogenetic group, closely related to Lactobacillus animalis , Lactobacillus apodemi and Lactobacillus murinus , by phylogenetic analysis based on 16S rRNA and recA gene sequences. Levels of DNA–DNA relatedness revealed that the isolates belonged to the same taxon and were genetically separated from their phylogenetic relatives. The three strains were non-motile, obligately homofermentative and produced l-lactic acid as the main end-product from d-glucose. The strains metabolized raffinose. The major cellular fatty acids in the three strains were C16 : 0, C18 : 1ω9c and C19 : 1 cyclo 9,10. Based on the data provided, it is concluded that the three strains represent a novel species of the genus Lactobacillus , for which the name Lactobacillus faecis sp. nov. is proposed. The type strain is AFL13-2T ( = JCM 17300T = DSM 23956T).

A novel Gram-stain-positive, facultatively anaerobic, non-motile and lactic-acid-producing bacterium, designated strain ORL-24T, was isolated from the gut of the camel cricket, Diestrammena coreana. Optimal growth occurred at 37 °C, pH 8 and with 0 % (w/v) NaCl. The ratio of l-lactate to d-lactate in strain ORL-24T was 96 : 4. Lancefield antigen D was not detected. The strain was negative for oxidase activity and catalase activity. According to a phylogenetic analysis based on 16S rRNA gene sequences, strain ORL-24T was most closely related to the type strain of Enterococcus asini (96.9 % similarity). Comparative pheS and rpoA sequence analyses of strain ORL-24T indicated that the strain belonged to the genus Enterococcus . The major fatty acids were C16 : 0 and C18 : 1ω9c. The DNA G+C content was 41.3 mol%. Based on phenotypic, genotypic and phylogenetic analyses, strain ORL-24T represents a novel species of the genus Enterococcus , for which the name Enterococcus diestrammenae is proposed. The type strain is ORL-24T ( = KACC 16708T = JCM 18359T).

A novel filamentous bacterium, strain SCSIO 11153T, was isolated from a sediment sample collected from the Indian Ocean (80° 03.099′ E 01° 03.300′ N) at a depth of 4593 m. Good growth was observed at 50–55 °C and pH 7.0 with 3 % NaCl. It formed ivory–white colonies with radial wrinkles. Aerial mycelium was absent on the media tested. Phenotypic characteristics and 16S rRNA gene sequence analysis indicated that strain SCSIO 11153T belonged to the family Thermoactinomycetaceae . It exhibited 96.4 % and 96.2 % 16S rRNA gene sequence similarities to Melghirimyces algeriensis NariEXT and Melghirimyces thermohalophilus Nari11AT, respectively, while lower sequence similarity values (<95.4 %) were observed between strain SCSIO 11153T and other species of genera in the family Thermoactinomycetaceae . The menaquinone type was MK-7. Major cellular fatty acids were iso-C15 : 0, anteiso-C15 : 0 and iso-C17 : 0. The polar lipids were diphosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylethanolamine and phosphatidylglycerol. The DNA G+C content of strain SCSIO 11153T was 52.6 mol%. On the basis of the genotypic and phenotypic characteristics, it is proposed that strain SCSIO 11153T represents a novel species of the genus Melghirimyces with the name Melghirimyces profundicolus sp. nov. The type strain is SCSIO 11153T ( = DSM 45787T = CCTCC AA 2012007T = NBRC 109068T).

A novel Gram-stain-positive bacterium, designated strain YIM M11385T, was isolated from a marine sediment sample collected from the South Bay, Little Andaman Island, India with a salinity of 35 p.p.m., pH 8.5. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain YIM M11385T belongs to the genus Marininema , supported by a bootstrap value of 100 %. The taxonomic position of this organism was further established by using a polyphasic approach. Strain YIM M11385T grew optimally at 28 °C, pH 7.0 and in the presence of 0–5 % (w/v) NaCl. The 16S rRNA gene sequence similarity between strain YIM M11385T and Marininema mesophilum SCSIO 10219T was 98.3 %. Strain YIM M11385T exhibited a quinone system with only MK-7, the polar lipid profile included diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine as major components, and the major fatty acids were anteiso-C15 : 0, iso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 0. The level of DNA–DNA relatedness between strain YIM M11385T and M. mesophilum SCSIO 10219T was 59.36 %. On the basis of genotypic and phenotypic data, it is apparent that strain YIM M11385T represents a novel species of the genus Marininema , for which the name Marininema halotolerans sp. nov. is proposed. The type strain is YIM M11385T ( = CCTCC AB 2012052T = DSM 45789T). In addition, we propose that the description of the genus Marininema should be further emended based on the results of the present study.