- Volume 66, Issue 9, 2016

The taxonomic position of the mountain soil actinomycete, strain CYP1-1BT, was clarified by a polyphasic study. The strain produced a single spore, or occasionally a chain of spores, on aerial mycelium. Chemotaxonomic data supported the classification of CYP1-1BT as representing a member of the genus Actinomadura on the basis of the presence of meso-diaminopimelic acid in the peptidoglycan; galactose, glucose, madurose and ribose as whole cell sugars; MK-9(H6), MK-9(H8) and MK-9(H4) as dominant menaquinones; C16 : 0, 10-methylated C18 : 0 and C18 : 1ω9c as the major cellular fatty acids; and diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and phosphatidylinositol manosides as the predominant phospholipids. The DNA G+C content was 74.3 mol%. On the basis of the combination of morphological and chemotaxonomic characteristics, CYP1-1BT was identified as representing a member of the genus Actinomadura . On the basis of the results of 16S rRNA gene analysis, CYP1-1BT, was shown to be closely related to Actinomadura nitritigenes DSM 44137T (98.9 %). Phenotypic, genotypic and DNA–DNA hybridization data supported the hypothesis that CYP1-1BT represents a novel species of the genus Actinomadura for which the name Actinomadura montaniterrae sp. nov. is proposed. The type strain is CYP1-1BT (=JCM 16995T=KCTC 39784T=PCU 349T=TISTR 2400T).

A nocardioform strain IFM 11456T was isolated from the aqueous humor from a patient with endophthalmitis and was characterized to its taxonomic position. IFM 11456T contained arabinose, galactose and meso-diaminopimelic acid in whole-cell hydrolysates and mycolic acids that co-migrated with those from the type strain of Nocardia asteroides . The acyl type of muramic acid was N-glycolyl. The diagnostic polar lipids were phosphatidylethanolamine, diphosphatidylglycerol and two unidentified glycolipids and the predominant menaquinone was MK-8(H4, ω-cycl.). These characteristics are typical of members of the genus Nocardia . Results of phylogenetic analyses based on 16S rRNA gene sequences indicated that the isolate represented a novel species of the genus Nocardia and was most closely related to the type strains of Nocardia mikamii JCM 15508T (98.1 %) and Nocardia aobensis IFM 0372T (98.1 %). However, analysis of partial gyrB sequences showed that strain IFM 11456T had 90.2 % similarity to Nocardia concava IFM 0354T and 90 % to Nocardia niigatensis IFM 0330T. The DNA–DNA relatedness values for strain IFM 11456T compared with N. mikamii JCM 15508T, N. aobensis IFM 0372T and N. concava IFM 0354T ranged from 24.4 to 39.9 %. Phenotypic characteristics that differentiated IFM 11456T from phylogenetically related species were growth at 45 °C, utilization of citrate and growth with inositol as a sole carbon source. On the basis of this polyphasic study, the isolate represents a novel species within the genus Nocardia , for which the name Nocardia shinanonensis sp. nov. is proposed. The type strain is IFM 11456T (=NBRC 109590T=TBRC 5149T).

Two bacterial strains, HKU54T and HKU55, were isolated from the oral cavity of two Chinese cobras (Naja atra) in Hong Kong. 16S rRNA gene sequence analysis revealed 100 % sequence identity between HKU54T and HKU55, and the two strains shared 99.0 % sequence identities with T sukamurella inchonensis ATCC 700082T. The two strains had unique biochemical profiles distinguishable from closely related species of the genus Tsukamurella . DNA–DNA hybridization confirmed that they belonged to the same species (≥92.1±7.9 % DNA–DNA relatedness) but were distinct from all other known species of the genus Tsukamurella (≤52.6±5.3 % DNA–DNA relatedness). Chemotaxonomic and morphological analyses of the two strains also demonstrated results consistent with their classification in the genus Tsukamurella . The DNA G+C contents of strains HKU54T and HKU55 were 69.2±1.5 mol% and 69.2±1.3 mol% (mean±sd; n=3) respectively. A novel species, Tsukamurella serpentis sp. nov., is proposed to accommodate strains HKU54T and HKU55, with HKU54T (=JCM 31017T=DSM 100915T) designated as the type strain.

A Gram-reaction-positive, aerobic, non-motile, irregular coccoid strain, designated YIM 7505T, was isolated from a leaf of Sweet Basil. Phylogenetic analysis on the basis of 16S rRNA gene sequence comparisons revealed that strain YIM 7505T was closely related to Flexivirga alba NBRC 107580T (98.9 % 16S rRNA gene sequence similarity) and formed a robust clade with F. alba NBRC 107580T in the neighbour-joining tree. Optimum growth of strain YIM 7505T was observed at 28–35 °C, pH 7.0 and in the presence of 0–3.0 % NaCl (w/v). The chemotaxonomic profiles of the strain comprised of anteiso-C16 : 0 as the major cellular fatty acid and MK-8(H4) as the respiratory menaquinone. The peptidoglycan of strain YIM 7505T contained serine, alanine, glycine, glutamic acid and lysine. The polar lipids contained diphosphatidylglycerol, phosphatidylglycerol, six unidentified phospholipids, four unidentified glycolipids, an unidentified aminolipid and an unidentified aminophospholipid. The G+C contents of the genomic DNA of strain YIM 7505T was 66.7 mol%. DNA–DNA hybridizations of strain YIM 7505T with F. alba NBRC 107580T gave relatedness values of 50.6±2.2 %. On the basis of the data recorded from the present study, strain YIM 7505T is considered to represent a novel species of the genus Flexivirga, for which the name Flexivirga endophytica sp. nov. is proposed. The type strain is YIM 7505T (=KCTC 39536T=CGMCC 1.15085T).

A polyphasic taxonomic study was performed on two Gram-positive-staining, anaerobic, pleomorphic, rod-shaped strains isolated from human bone and tissue samples. Sequencing of the 16S rRNA genes revealed that the strains belong to a novel species within the genus Propionibacterium , most closely related to Propionibacterium acnes subsp. acnes and Propionibacterium acnes subsp. elongatum with similarity values of 98.4 % and 98.1 %, respectively. In addition, protein-coding genes for rpoB, recA and gyrB clearly separated the novel organism from all species and subspecies of the genus Propionibacterium . However, a DNA–DNA hybridization analysis between the novel organism and the type strain P. acnes ATCC 6919T revealed a value of only 61.1 %. Furthermore, whole genome analysis using the program OrthoANI gave a value of 88.5 %, which is significantly below the cut-off value of 95 % for species delineation. The major fatty acids were iso-C15 : 0, anteiso-C15 : 0 and iso-C17 : 0. The DNA G+C content of the type strain was 59.7 mol%. When taken collectively, phenotypic, molecular genetic, chemotaxonomic and phylogenetic information demonstrate that the organism represents a distinct, albeit close relative of P. acnes On the basis of the results presented, the organism represents a novel member of the genus Propionibacterium for which the name Propionibacterium namnetense sp. nov. is proposed. The type strain is NTS 31307302T (=DSM 29427T=CCUG 66358T).

A diversity study on the presence of strains representing the genus Micromonospora in Pisum sativum nodules collected from Cañizal (Spain) has provided evidence of the high number of isolates that might represent novel species. In the present work, we have characterized three of these isolates: GUI23T, GUI43T and GUI63T. Phenotypic and genotypic analyses confirmed that all strains represent novel species of the genus Micromonospora with the following proposed names: Micromonospora ureilytica sp. nov., type strain GUI23T (=CECT 9022T=DSM 101692T), Micromonospora noduli sp. nov., type strain GUI43T (=CECT 9020T=DSM 101694T), and Micromonospora vinacea sp. nov., type strain GUI63T (=CECT 9019T=DSM 101695T).

A Gram-stain-positive, alkaliphilic, non-spore-forming, non-motile bacterium, designated VDS11T, was isolated from a soil sample collected from the hexachlorocyclohexane dumpsite, located at Ummari Village, Lucknow, Uttar Pradesh, India. 16S rRNA gene sequence analysis indicated that strain VDS11T occupies a distinct phylogenetic position within the genus Corynebacterium , showing the highest sequence similarity with Corynebacterium humireducens MFC-5T (98.7 %) and Corynebacterium nasicanis 2673/12T (98.4 %). The DNA G+C content was 50.6 mol% and the DNA–DNA hybridization (DDH) relatedness value with C. humireducens MFC-5T and C. nasicanis 2673/12T was 49.7 and 39.6 %, respectively. Strain VDS11T contained C16 : 0 (28.3), C18 : 1ω9c (52.3), C18 : 0 (1.3) and C20 : 0 (3.0) as the major cellular fatty acids. The major isoprenoid quinone was MK-9(H2). Strain VDS11T contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, unidentified aminolipids, glycolipids, phospholipids and unidentified lipids as the major polar lipids. The peptidoglycan type was A31 (meso 2, 6-diaminopimelic acid, directly cross-linked). Based on the results of DDH studies and the biochemical and physiological data, strain VDS11T represents a novel species of the genus Corynebacteriumfor which the name Corynebacterium pollutisoli sp. nov., has been proposed. The type strain is VDS11T(=DSM 100104T=MCC 2722T=KCTC 39687T).

Three novel isolates belonging to the genus Streptomyces , designated JR-35T, JR-46 and WH-9T, were isolated from bamboo forest soil in Damyang, Korea. The 16S rRNA gene sequences of strains JR-35T and JR-46 showed highest similarities with Streptomyces olivochromogenes NBRC 3178T (99.1 %), Streptomyces siamensis KC-038T (98.9 %), Streptomyces chartreusis NBRC 12753T (98.9 %), Streptomyces resistomycificus NRRL ISP-5133T (98.9 %) and Streptomyces bobili JCM 4627T (98.8 %), and strain WH-9Tshowed highest sequence similarities with Streptomyces . bobili JCM 4627T (99.2 %), Streptomyces phaeoluteigriseus NRRL ISP-5182T (99.2 %), Streptomyces alboniger NBRC 12738T (99.2 %), Streptomyces galilaeus JCM 4757T (99.1 %) and Streptomyces pseudovenezuelae NBRC 12904T (99.1 %). The predominant menaquinones were MK-9 (H6) and MK-9 (H8). The major fatty acids were anteiso-C15 : 0, iso-C16 : 0, iso-C14 : 0 and iso-C15 : 0 for strains JR-35T and JR-46 and anteiso-C15 : 0, iso-C15 : 0 and iso-C16 : 0 for strain WH-9T. The G+C content of the genomic DNA of strains JR-35T, JR-46 and WH-9T were 69.4, 74.4 and 74.1 mol%, respectively. Based on the phenotypic and genotypic data, the three strains are assigned to two novel species of the genus Streptomyces , for which the names Streptomyces rhizosphaerihabitans sp. nov. (type stain JR-35T=KACC 17181T=NBRC 109807T) and Streptomyces adustus sp. nov. (type strain WH-9T=KACC 17197T=NBRC 109810T) are proposed.

A novel Gram-staining-positive, aerobic, non-motile and coccus-shaped bacterium, designated strain TBS-100T, was isolated from the faeces of a crested ibis, Nipponia nippon. The phylogenetic analysis based on the 16S rRNA gene sequences showed that the closest relative of TBS-100T was Flexivirga alba DSM 24460T with 97.11 % sequence similarity, and that strain TBS-100T belonged to the genus Flexivirga . The optimum growth conditions for strain TBS-100T were 30 °C, at a pH of 7 and in the presence of 0 % (w/v) NaCl. The primary cellular fatty acids of strain TBS-100T were anteiso-C17 : 0 and iso-C17 : 0. The predominant isoprenoid quinones were MK-8 (H4) (70.2 %) and MK-8 (H6) (29.7 %). The polar lipids were diphosphatidylglycerol, phosphatidylinositol, seven unidentified lipids and an unidentified phospholipid. The whole-cell sugars of strain TBS-100T were ribose, glucose, galactose, rhamnose and mannose. The peptidoglycan contained alanine, lysine, glutamic acid, glycine and aspartic acid. The DNA G+C content was 64.8 mol%. The phenotypic, phylogenetic and genotypic analyses indicated that strain TBS-100T represents a novel species of the genus Flexivirga for which the name Flexivirga lutea sp. nov. is proposed. The type strain is TBS-100T (=KCTC 39625T=JCM 31200T). In addition, an emended description of the genus Flexivirga is proposed.

Five actinomycete isolates, CPB1-1T, CPB2-10, BM1-4, CPB3-1 and CPB1-18, belonging to the genus Streptomyces were isolated from marine sediments collected from Chumphon Province, Thailand. They produced open loops of warty spore chains on aerial mycelia. ll-Diaminopimelic acid, glucose and ribose were found in their whole-cell hydrolysates. Polar lipids found were diphosphatidylglycerol, phosphatidylethanolamine, lysophosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannoside. Menaquinones were MK-9(H6), MK-9(H8), MK-10(H6) and MK-10(H8). Major cellular fatty acids were anteiso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 0. The taxonomic position of the strains was described using a polyphasic approach. blastn analysis of the 16S rRNA gene sequence revealed that these five strains exhibited the highest similarities with ‘ Streptomyces mangrovicola ' GY1 (99.0 %), Streptomyces fenghuangensis GIMN4.003T (98.6 %), Streptomyces barkulensis RC 1831T (98.5 %) and Streptomyces radiopugnans R97T (98.3 %). However, their phenotypic characteristics and 16S rRNA gene sequences as well as DNA–DNA relatedness differentiated these five strains from the other species of the genus Streptomyces . Here, we propose the novel actinomycetes all being representatives of the same novel species, Streptomyces verrucosisporus , with type strain CPB1-1T (=JCM 18519T=PCU 343T=TISTR 2344T).

An actinobacterium, designated strain CPCC 203993T, was isolated from a rhizosphere soil sample collected from Heilongjiang Province, northeast China, and was characterized using a polyphasic taxonomy approach. Cells of the strain were Gram-stain-positive, non-motile and non-endospore-forming cocci. The 16S rRNA gene sequence comparison of strain CPCC 203993T with members of the genus Microlunatu s yielded 93.9 % to 97.8 % similarities. In the phylogenetic tree based on 16S rRNA gene sequences, strain CPCC 203993T was affiliated to the clade of the genus Microlunatus next to Microlunatus parietis DSM 22083T, while the DNA–DNA hybridization value of 31.5 % (±1.8 %) between strain CPCC 203993T and Microlunatus. parietis DSM 22083T was far below 70 %. This result indicated that strain CPCC 203993T represented a different genomic species from M. parietis . Chemotaxonomically, the strain contained ll-2,6-diaminopimelic acid as the diagnostic diamino acid, MK-9(H4) as the only menaquinone, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, three unidentified glycolipids and one unidentified phospholipid in the polar lipids extracts, and anteiso-C15 : 0, iso-C15 : 0 and iso-C16 : 0 as the major cellular fatty acids, without mycolic acids. The genomic DNA G+C content was 64.04 mol%. The above evidence from the polyphasic study merit the recognition of strain CPCC 203993T as a representative of a novel species of the genus Microlunatus , for which Microlunatus nigridraconis sp. nov. is proposed. The type strain is CPCC 203993T (=DSM 29529T=NBRC 110715T=KCTC 29689T).

From our recent survey of non-pigmented rapidly growing mycobacteria in the Parisian water system, three groups of isolates (taxons 1–3) corresponding to possible novel species were selected for taxonomic study. The three taxa each formed creamy white, rough colonies, had an optimal growth temperature of 30 °C, hydrolyzed Tween 80, were catalase-positive at 22 °C and expressed arylsulfatase activity. All three were susceptible to amikacin, ciprofloxacin and tigecycline. The three taxa produced specific sets of mycolic acids, including one family that has never previously been described, as determined by thin layer chromatography and nuclear magnetic resonance. The partial rpoB sequences (723 bp) showed 4–6 % divergence from each other and more than 5 % differences from the most similar species. Partial 16S rRNA gene sequences showed 99 % identity within each species. The most similar sequences for 16S rRNA genes (98–99 % identity over 1444–1461 bp) were found in the Mycobacterium fortuitum group, Mycobacterium septicum and Mycobacterium farcinogenes . The three taxa formed a new clade (bootstrap value, 99 %) on trees reconstructed from concatenated partial 16S rRNA, hsp65 and rpoB sequences. The above results led us to propose three novel species for the three groups of isolates, namely Mycobacterium lutetiense sp. nov. [type strain 071T=ParisRGMnew_1T (CIP 110656T=DSM 46713T)], Mycobacterium montmartrense sp. nov. [type strain 196T=ParisRGMnew_2T (CIP 110655T=DSM 46714T)] and Mycobacteriu marcueilense sp. nov. [type strain of 269T=ParisRGMnew_3T (CIP 110654T=DSM 46715T)].

Four Gram-stain-positive, aerobic, motile bacterial strains were isolated from the bark tissue of Populus × euramericana canker. Growth occurred between 10 and 37 °C and at pH 6–10, with optimal growth at 28–30 °C and pH 7.0–8.0. Growth occurred at 0–3 % (w/v) salinity. The strains were positive for oxidase and catalase activity. The major fatty acids were anteiso-C15 : 0 and C16 : 0. The phospholipid profiles contained diphosphatidylglycerol, phosphatidylinositol, phosphatidylglycerol, two phospholipids and five glycolipids. The peptidoglycan type was A4α, which is based on l-Lys–d-Ser–d-Asp. The DNA G+C content was 58.5 mol%. Based on 16S rRNA gene sequence analysis, as well as physiological and biochemical characteristics, the strains are considered to represent a novel species of a new genus in the family Jonesiaceae . The name proposed is Populibacterium corticicola gen. nov., sp. nov. The type strain of Populibacterium corticicola is 2D-4T (=CFCC 11886T=KCTC 33576T).

A Gram-reaction-positive, motile, yellow-pigmented and rod-shaped bacterial strain, designated AR33T, was isolated from the rhizosphere of Salix caprea L. growing in a former zinc/lead mining and processing site in Austria. A polyphasic approach was applied to determine its taxonomic position. 16S rRNA gene sequence analysis, and morphological and chemotaxonomic properties showed that strain AR33T belongs to the genus Agromyces . Strain AR33T had peptidoglycan type B2γ and the major menaquinones were MK-11, MK-10 and MK-12. The main branched-chain fatty acids were anteiso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 0. Strain AR33T showed catalase and oxidase activity and multiple heavy metal resistances to zinc, lead and cadmium. The DNA G+C content was 70.1 mol%. Levels of 16S rRNA gene sequence similarity with closely related recognized species of the genus Agromyces ranged between 98 and 99 %. However, DNA–DNA hybridization between strain AR33T and the type strains of three Agromyces species showed values lower than 42 % relatedness. Therefore, differential phenotypic characteristics together with DNA–DNA relatedness suggested that strain AR33T can be recognized as representing a distinct Agromyces species, for which the name Agromyces aureus sp. nov. is proposed. The type strain is AR33T (=DSM 101731T=LMG 29235T).

A hyperthermophilic, autotrophic iron and nitrate reducer, strain Su06T, was isolated from an active deep-sea hydrothermal vent chimney on the Endeavour Segment in the north-eastern Pacific Ocean. It was obligately anaerobic, hydrogenotrophic and reduced Fe(III) oxide to magnetite and NO3 − to N2. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the strain was more than 97 % similar to other species of the genera Pyrodictium and Hyperthermus . Therefore, overall genome relatedness index analyses were performed to establish whether strain Su06T represents a novel species. For each analysis, strain Su06T was most similar to Pyrodictium occultum PL-19T. Relative to this strain, the average nucleotide identity score for strain Su06T was 72 %, the genome-to-genome direct comparison score was 13–19 % and the species identification score at the protein level was 89 %. For each analysis, strain Su06T was below the species delineation cutoff. Based on its whole genome sequence and its unique phenotypic characteristics, strain Su06T is suggested to represent a novel species of the genus Pyrodictium , for which the name Pyrodictium delaneyi is proposed. The type strain is Su06T (=DSM 28599T=ATCC BAA-2559T).

A Gram-staining-negative, rod-shaped, facultatively anaerobic, non-motile and pale-red-pigmented bacterium, designated SAORIC-476T, was isolated from deep-sea water from the Pacific Ocean. 16S rRNA gene sequence analyses showed that strain SAORIC-476T was most closely related to Rubrivirga marina SAORIC-28T (96.8 % similarity) and formed a robust phylogenetic clade with Rubrivirga marina of the family Rhodothermaceae . Optimal growth of strain SAORIC-476T was observed at 25 °C, pH 7.5 and in the presence of 3.0 % (w/v) NaCl. The DNA G+C content of strain SAORIC-476T was 66.2 mol%, and the sole isoprenoid quinone was MK-7. The predominant cellular fatty acids were summed feature 9 (iso-C17 : 1ω9c and/or 10-methyl C16 : 0), iso-C17 : 0, C17 : 1ω8c and iso-C15 : 0. The major polar lipids constituted phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, three unknown phospholipids and four unknown polar lipids. On the basis of taxonomic data obtained in this study, it was concluded that strain SAORIC-476T represents a novel species of the genus Rubrivirga , for which the name Rubrivirga profundi sp. nov. is proposed. The type strain of Rubrivirga profundi is SAORIC-476T (=NBRC 110607T=KACC 18401T).

A Gram-staining-negative, strictly aerobic and yellow-pigmented bacterium, designated strain S2-22T, was isolated from estuary sediment in South Korea. Cells of strain S2-22T were oxidase- and catalase-positive rods without gliding motility. Growth was observed at 15–43 °C (optimum, 35–37 °C), at pH 5.5–9.0 (optimum, pH 6.5–7.5) and in the presence of 0.0–10.0 % (w/v) NaCl (optimum, 2.0 %). Phylogenetic analysis based on 16S rRNA gene sequences showed that strain S2-22T formed a tight phyletic lineage with Zeaxanthinibacter enoshimensis TD-ZE3T with a high bootstrap value and their 16S rRNA gene sequence similarity was 94.6 %. The respiratory quinone detected was menaquinone 6 (MK-6) only and iso-C15 : 0, iso-C17 : 0 3-OH, summed feature 3 (comprising C16 : 1ω7c/C16 : 1ω6c) and iso-C15 : 1G were the major fatty acids. Phosphatidylethanolamine was identified as the major polar lipid and five unidentified aminolipids, two unidentified phospholipids and three unidentified lipids were also detected as minor polar lipids. The G+C content of the genomic DNA was 45.5 mol%. Based on the phenotypic, chemotaxonomic and molecular features, strain S2-22T clearly represents a novel species of the genus Zeaxanthinibacter , for which the name Zeaxanthinibacter aestuarii sp. nov. is proposed. The type strain is S2-22T (=KACC 18503T=JCM 31155T). An emended description of the genus Zeaxanthinibacter is also proposed.