- Volume 65, Issue Pt_7, 2015

A Gram-stain-positive, facultatively anaerobic, non-spore-forming, non-flagellated and rod-shaped or ovoid bacterial strain, designated BS-12MT, was isolated from a tidal flat sediment on the South Sea, South Korea. Strain BS-12MT grew optimally at 35 °C, at pH 7.0–8.0 and in the presence of 2.0 % (w/v) NaCl. The neighbour-joining phylogenetic tree based on 16S rRNA gene sequences revealed that strain BS-12MT fell within the cluster comprising the type strains of species of the genus Demequina, joining the type strain of Demequina aestuarii with which it shared the highest sequence similarity (98.6 %). It exhibited 16S rRNA gene sequence similarity of 96.1–97.9 % to the type strains of other species of the genus Demequina. The peptidoglycan type of strain BS-12MT was A4β based on l-Orn − l-Ser − d-Glu. Strain BS-12MT contained demethylmenaquinone-9(H4) as the major menaquinone and anteiso-C15:0 and C16:0 as the major fatty acids. The major polar lipids of strain BS-12MT were phosphatidylinositol and phosphatidylinositolmannoside. The DNA G+C content of strain BS-12MT was 70.7 mol% and its DNA–DNA relatedness values with the type strains of five phylogenetically related species of the genus Demequina were 15–34 %. Differential phenotypic properties, together with phylogenetic and genetic distinctiveness, revealed that strain BS-12MT is separate from other species of the genus Demequina. On the basis of the data presented, strain BS-12MT is considered to represent a novel species of the genus Demequina, for which the name Demequina activiva sp. nov. is proposed. The type strain is BS-12MT ( = KCTC 29674T = NBRC 110675T).

A novel bacterium (strain NIO-1002T) belonging to the genus Microbacterium was isolated from a marine sediment sample in Chorao Island, Goa Province, India. Its morphology, physiology, biochemical features and 16S rRNA gene sequence were characterized. Cells of this strain were Gram-stain-positive, non-motile, non-spore-forming rods that formed yellow-pigmented colonies. It grew in 0–12 % (w/v) NaCl and at 25–37 °C, with optimal growth at 30 °C. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain NIO-1002T is associated with members of the genus Microbacterium, with highest sequence similarity with Microbacterium hominis CIP 105731T (98.1 %) and Microbacterium testaceum KCTC 9103T (98.0 %). Within the phylogenetic tree, this novel strain shared a branching point with M. hominis CIP 105731T. The DNA G+C content was 66.5 mol% and DNA–DNA hybridization relatedness between NIO-1002T, M. hominis CIP 105731T and M. testaceum KCTC 9103T was 39.0 ± 2.0 % and 41.0 ± 2.0 %, respectively. The major fatty acids were ai-C15 : 0, i-C16 : 0 and ai-C17 : 0 and the diamino acid in the cell-wall peptidoglycan of NIO-1002T was lysine. Data obtained from DNA–DNA hybridization and chemotaxonomic phenotypic analysis support the conclusion that strain NIO-1002T represents a novel species within the genus Microbacterium. The name Microbacterium enclense sp. nov. is proposed, with NIO-1002T ( = NCIM 5454T = DSM 25125T = CCTCC AB 2011120T) as the type strain.

Two isolates from water, D16Q19 and D16R27, were shown to be highly similar in their 16S rRNA, 16S–23S internal transcribed spacer (ITS), hsp65 and rpoB gene sequences to ‘Mycobacterium franklinii’ DSM 45524, described in 2011 but with the name not validly published. They are all nonpigmented rapid growers and are related phenotypically and genetically to the Mycobacterium chelonae–Mycobacterium abscessus group. Extensive characterization by phenotypic analysis, biochemical tests, drug susceptibility testing, PCR restriction enzyme analysis of the hsp65 gene and ITS, DNA sequencing of housekeeping genes and DNA–DNA hybridization demonstrated that ‘M. franklinii’ DSM 45524, D16Q19 and D16R27 belong to a single species that is separated from other members of the M. chelonae–M. abscessus group. On the basis of these results we propose the formal recognition of Mycobacterium franklinii sp. nov. Strain DSM 45524T ( = ATCC BAA-2149T) is the type strain.

A Gram-staining-positive, white-coloured, aerobic, non-motile, catalase-positive and oxidase-negative, endophytic actinobacterium, designated strain EGI 6500322T, was isolated from the surface-sterilized root of the halophyte Salsola affinis C. A. Mey collected from Urumqi, Xinjiang province, north-west China. Growth occurred at 5–35 °C (optimum 25–30 °C), at pH 5–10 (optimum pH 7–8) and with 0–13 % NaCl (w/v) (optimum 0–5 %). The predominant menaquinone was MK-9 (93.1 %). The major cellular fatty acids were anteiso-C15:0 (49.5 %) and iso-C15:0 (15.1 %). The cell-wall peptidoglycan contained lysine, alanine and glutamic acid. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, two unknown phospholipids and one unknown glycolipid. The DNA G+C content of strain EGI 6500322T was 62.0 mol%. Based on 16S rRNA gene sequence analysis, the nearest phylogenetic neighbours of strain EGI 6500322T were identified as Arthrobacter ardleyensis DSM 17432T (98.38 %) and Arthrobacter bergerei DSM 16367T (98.37 %). The DNA–DNA relatedness between strain EGI 6500322T and Arthrobacter ardleyensis DSM 17432T and Arthrobacter bergerei DSM 16367T was 53.4 ± 4.1 % and 30.5 ± 1.7 %, respectively. On the basis of the phylogenetic analysis, chemotaxonomic data, physiological characteristics and DNA–DNA hybridization data, strain EGI 6500322T should represent a novel species of the genus Arthrobacter, for which the name Arthrobacter endophyticus sp. nov. is proposed. The type strain is EGI 6500322T ( = KCTC 29490T = JCM 30091T).

From the whole blood of Korean native cattle, Hanwoo (Bos taurus coreanae), a previously undescribed, rapidly growing, scotochromogenic isolate of the genus Mycobacterium is reported. Its 16S rRNA gene sequence, and the sequences of three other genes (hsp65, recA and rpoB) were unique and phylogenetic analysis based on 16S rRNA gene sequence (1420 bp) placed the organism into the rapidly growing Mycobacterium group close to Mycobacterium smegmatis (98.5 % sequence similarity). However, phylogenetic analyses based on three different gene sequences (hsp65, recA and rpoB) revealed its location to be distinct from the branch of rapidly growing species. Culture and biochemical characteristics were generally similar to those of Mycobacterium fortuitum. Unique matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) MS profiles of lipids, unique fatty acid profile, unique mycolic acids profiles and a low DNA–DNA relatedness to M. fortuitum (23.6 %) and M. smegmatis (39.7 %) strongly supported the taxonomic status of this strain as a representative of a novel species of rapidly growing mycobacteria named Mycobacterium anyangense. The type strain is strain QIA-38T ( = JCM 30275T = KCTC 29443T).

A novel marine actinomycete, designated B374T, was isolated from a marine sponge, Glodia corticostylifera, which was collected from São Paulo, Brasil. The taxonomic position of B374T was established by using data derived from a polyphasic approach. The organism showed a combination of chemotaxonomic and morphological characteristics consistent with its classification in the genus Marmoricola and it formed a distinct phyletic line in the clade of the genus Marmoricola, based on 16S rRNA gene sequences. Strain B374T was most closely related to Marmoricola aequoreus SST-45T (98.5 % 16S rRNA gene sequence similarity), but was distinguished from this strain and from the other type strains of species of the genus Marmoricola on the basis of a combination of phenotypic properties. The data obtained, therefore, indicates that isolate B374T ( = CBMAI 1089T = DSM 28169T) should be classified as a novel species of the genus Marmoricola, for which the name Marmoricola aquaticus sp. nov. is proposed.

A Gram-staining-positive, rod-shaped, endospore-forming, aerobic bacterium (FJAT-17212T) was isolated from the rhizosphere soil of a medical plant, Prunella vulgaris (common selfheal), on the Wuyishan mountain of China. Isolate FJAT-17212T grew at 10–50 °C (optimum 30 °C), pH 5–11 (optimum pH 7) and with 0–6  % (w/v) NaCl (optimum 2  %). Phylogenetic analyses based on 16S rRNA gene sequences showed that isolate FJAT-17212T was a member of the genus Bacillus and was most closely related to Bacillus galactosidilyticus DSM 15595T (97.3  %). DNA–DNA relatedness between isolate FJAT-17212T and B. galactosidilyticus DSM 15595T was low (35.2  % ± 2.3). The diagnostic diamino acid of the peptidoglycan of isolate FJAT-17212T was meso-diaminopimelic acid and the predominant isoprenoid quinone was MK-7 (80.8  %). The major cellular fatty acids were iso-C15 : 0 (35.7  %), anteiso-C15 : 0 (29.8  %), iso-C14 : 0 (9.9  %) and iso-C16 : 0 (9.9  %) and the DNA G+C content was 39.8  mol%. Phenotypic, chemotaxonomic and genotypic properties clearly indicated that isolate FJAT-17212T represents a novel species within the genus Bacillus, for which the name Bacillus wuyishanensis sp. nov. is proposed. The type strain is FJAT-17212T ( = DSM 27848T = CGMCC 1.12709T).

Seven coagulase-negative, oxidase-negative and novobiocin-susceptible staphylococci assigned tentatively as Staphylococcus petrasii were investigated in this study in order to elucidate their taxonomic position. All strains were initially shown to form a genetically homogeneous group separated from remaining species of the genus Staphylococcus by using a repetitive sequence-based PCR fingerprinting with the (GTG)5 primer. Phylogenetic analysis based on 16S rRNA gene, hsp60, rpoB, dnaJ, gap and tuf sequences showed that the group is closely related to Staphylococcus petrasii but separated from the three hitherto known subspecies, S. petrasii subsp. petrasii, S. petrasii subsp. croceilyticus and S. petrasii subsp. jettensis. Further investigation using automated ribotyping, MALDI-TOF mass spectrometry, fatty acid methyl ester analysis, DNA–DNA hybridization and extensive biotyping confirmed that the analysed group represents a novel subspecies within S. petrasii, for which the name Staphylococcus petrasii subsp. pragensis subsp. nov. is proposed. The type strain is NRL/St 12/356T ( = CCM 8529T = LMG 28327T).

A Gram-stain-positive, rod-shaped, endospore-forming, aerobic bacterium (FJAT-14571T) was isolated from a soil sample in Taiwan. Strain FJAT-14571T grew at 20–40 °C (optimum 35 °C), pH 6–10 (optimum pH 8) and 0–2 % (w/v) NaCl (optimum 0 %). Phylogenetic analyses based on 16S rRNA gene sequences showed that strain FJAT-14571T was a member of the genus Bacillus and was most closely related to Bacillus oceanisediminis DSM 24771T (96.2 %). DNA–DNA relatedness between strain FJAT-14571T and B. oceanisediminis DSM 24771T was low (32.0 % ± 0.88 %). The diagnostic diamino acid of the peptidoglycan of strain FJAT-14571T was meso-diaminopimelic acid and the predominant menaquinone was MK-7 (96.6 %). The major cellular fatty acids were iso-C15 : 0 (46.4 %), anteiso-C15 : 0 (7.6 %), iso-C17 : 0 (8.2 %) and iso-C16 : 0 (10.0 %) and the DNA G+C content was 40.8 mol%. Phenotypic, chemotaxonomic and genotypic properties clearly indicated that strain FJAT-14571T represents a novel species within the genus Bacillus, for which the name Bacillus taiwanensis sp. nov. is proposed. The type strain is FJAT-14571T ( = DSM 27845T = CGMCC1.1 2698T).

A Gram-stain-positive, rod-shaped, endospore-forming bacterium (designated strain CC-Bw-5T) was isolated from chopped tomato stems. The isolate grew at 20–40 °C, pH 6.0–8.0 and tolerated 6  % (w/v) NaCl. The most closely related strains in terms of 16S rRNA gene sequence similarity were Bacillus isabeliae (95.3  %) and Bacillus oleronius (95.3  %). The G+C content of the genomic DNA was 37.2 ± 3.6 mol%. Strain CC-Bw-5T was determined to possess C16 : 0, iso-C14 : 0, iso-C15 : 0, anteiso-C15 : 0 and iso-C16 : 0 as predominant fatty acids. The polar lipid profile consisted of predominant amounts of diphosphatidylglycerol, and moderate-to-trace amounts of phosphatidylethanolamine, phosphatidylglycerol, two unidentified glycolipids, an unidentified phospholipid and an unidentified phosphoglycolipid. The cell-wall peptidoglycan contained meso-diaminopimelic acid; menaquinone (MK-7) was the predominant respiratory quinone. According to distinct phylogenetic, phenotypic and chemotaxonomic features, strain CC-Bw-5T is proposed to represent a novel species within the genus Bacillus for which the name Bacillus lycopersici sp. nov. is proposed. The type strain is CC-Bw-5T ( = BCRC 80623T = JCM 19140T).

Two strains of lactic acid bacteria, designated 117T and 4195T, were isolated from goat milk in Valtellina, Italy and from cow milk in Valle Trompia, Italy, respectively, and characterized taxonomically by a polyphasic approach. The strains were Gram-stain-positive, coccoid, non-spore-forming and catalase-negative bacteria. Morphological, physiological and phylogenetic data indicated that these isolates belonged to the genus Lactococcus. Strain 117T was closely related to Lactococcus fujiensis, Lactococcus lactis subsp. lactis, L. lactis subsp. cremoris, L. lactis subsp. hordniae, L. lactis subsp. tructae and Lactococcus taiwanensis, showing 93–94 % and 82–89 % 16S rRNA and rpoB gene sequence similarities, respectively. Strain 4195T was closely related to Lactococcus chungangensis, Lactococcus raffinolactis, Lactococcus plantarum and Lactococcus piscium, showing 92–98  % and 86–99  % 16S rRNA and rpoB gene sequence similarities, respectively. Based on this evidence and the data obtained in the present study, the milk isolates represent two novel species of the genus Lactococcus, for which the names Lactococcus hircilactis sp. nov., and Lactococcus laudensis sp. nov. are proposed. The respective type strains are 117T ( = LMG 28352T = DSM 28960T) and 4195T ( = LMG 28353T = DSM 28961T).

The rhizosphere-isolated bacteria belonging to the Bacillus amyloliquefaciens subsp. plantarum and Bacillus methylotrophicus clades are an important group of strains that are used as plant growth promoters and antagonists of plant pathogens. These properties have made these strains the focus of commercial interest. Here, we present the draft genome sequence of B. methylotrophicus KACC 13105T ( = CBMB205T). Comparative genomic analysis showed only minor differences between this strain and the genome of the B. amyloliquefaciens subsp. plantarum type strain, with the genomes sharing approximately 95 % of the same genes. The results of morphological, physiological, chemotaxonomic and phylogenetic analyses indicate that the type strains of these two taxa are highly similar. In fact, our results show that the type strain of B. amyloliquefaciens subsp. plantarum FZB42T ( = DSM 23117T = BGSC 10A6T) does not cluster with other members of the B. amyloliquefaciens taxon. Instead, it clusters well within a clade of strains that are assigned to B. methylotrophicus, including the type strain of that species. Therefore, we propose that the subspecies B. amyloliquefaciens subsp. plantarum should be reclassified as a later heterotypic synonym of B. methylotrophicus.

A novel bacterial strain, S1-9T, was isolated from a lead–zinc tailing in Nanjing, Jiangsu Province, China. Cells of strain S1-9T were Gram-stain-negative, ellipsoidal endospore-forming, aerobic rods and motile by means of peritrichous flagella. On the basis of 16S rRNA gene sequence analysis, strain S1-9T was shown to belong to the genus Paenibacillus and the closest phylogenetic relatives were Paenibacillus glucanolyticus DSM 5162T (96.8 % similarity), Paenibacillus lautus NRRL NRS-666T (96.5 %) and Paenibacillus lactis MB 1871T (95.4 %). The predominant menaquinone was MK-7. The major cellular fatty acids were anteiso-C15:0 and iso-C16:0. The polar lipid profile contained phosphatidylglycerol, phosphatidylethanolamine, diphosphatidylglycerol, two unknown phospholipids and two unknown lipids. The total DNA G+C content of strain S1-9T was 49.9 mol%. Based on the low levels of DNA–DNA relatedness (ranging from 21.8 to 48.4 %) to the type strains of the above species of the genus Paenibacillus and unique phenotypic characteristics, strain S1-9T is considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus qingshengii sp. nov. is proposed. The type strain is S1-9T ( = CCTCC AB 2014290T = JCM 30613T).

A novel Gram-stain-positive, aerobic, endospore-forming, rod-shaped bacterial strain YN3T was isolated from ripened Pu'er tea. Phylogenetic analysis of 16S rRNA gene sequences showed that the strain belonged to the family Sporolactobacillaceae and was closely related to Pullulanibacillus naganoensis DSM 10191T (95.8 % 16S rRNA gene sequence similarity) and Pullulanibacillus uraniitolerans DSM 19429T (95.4 %). Growth of the strain was observed at 20–50 °C (optimum 30–37 °C), at pH 4.0–8.0 (optimum pH 5.0–6.0). The strain had a cell-wall type A1γ peptidoglycan with meso-diaminopimelic acid as the diagnostic diamino acid. The predominant menaquinone was menaquinone-7 (MK-7). The major fatty acids were anteiso-C15:0, anteiso-C17:0 and C18:1ω7c. The DNA G+C content of strain YN3T was 38.7 mol%. Strain YN3T could be differentiated from recognized species of the genus Pullulanibacillus based on phenotypic characteristics, chemotaxonomic differences, phylogenetic analysis and DNA–DNA hybridization data. On the basis of polyphasic evidence from this study, Pullulanibacillus pueri sp. nov., is proposed, with strain YN3T ( = CGMCC 1.12777T = JCM 30075T) as the type strain.

Bacterial strain THMBR28T was isolated from decomposing algal scum that was collected during an algal bloom in Taihu lake, China. Cells of strain THMBR28T were Gram-staining-positive, facultatively anaerobic and rod-shaped. Growth was observed at 20–45 °C (optimum, 30 °C), at pH 5.0–9.5 (optimum, pH 6.5–7.5), and in the presence of 0–1.0  % (w/v) NaCl (optimum, 0.5  %). Strain THMBR28T contained MK-7 as the major menaquinone and iso-C15 : 0 as the major cellular fatty acid. The polar lipid profile contained phosphatidylglycerol, phosphatidylmonomethylethanolamine, phosphatidylethanolamine and six unidentified polar lipids. The diamino acid found in the cell-wall peptidoglycan was meso-diaminopimelic acid. The DNA G+C content was 57.6 mol% (T m ). Phylogenetic analysis of 16S rRNA gene sequences showed that strain THMBR28T belonged to the genus Tumebacillus, most closely related to Tumebacillus ginsengisoli DSM 18389T (95.0  %) and Tumebacillus permanentifrigoris Eur1 9.5T (93.4  %). Based on phylogenetic and phenotypic characterization, it is concluded that strain THMBR28T represents a novel species of the genus Tumebacillus, for which the name Tumebacillus algifaecis sp. nov. is proposed, with THMBR28T ( = CGMCC 1.10949T = NBRC 108765T) as the type strain.

A Gram-stain-positive, endospore-forming, rod-shaped bacterial strain, designated D5T, was isolated from seawater collected from a sandy beach in a southern state of Malaysia and subjected to a polyphasic taxonomic study. Sequence analysis of the 16S rRNA gene demonstrated that this isolate belongs to the genus Jeotgalibacillus, with 99.87 % similarity to Jeotgalibacillus alimentarius JCM 10872T. DNA–DNA hybridization of strain D5T with J. alimentarius JCM 10872T demonstrated 26.3 % relatedness. The peptidoglycan type was A1α linked directly to l-lysine as the diamino acid. The predominant quinones identified in strain D5T were menaquinones MK-7 and MK-8.The major fatty acids were iso-C15:0 and anteiso-C15:0. The G+C content of its DNA was 43.0 mol%. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and sulfoquinovosyl diacylglycerol, as well as two unknown phospholipids and three unknown lipids. The phenotypic, chemotaxonomic and genotypic data indicated that strain D5T represents a novel species of the genus Jeotgalibacillus, for which the name Jeotgalibacillus malaysiensis sp. nov. is proposed (type strain D5T = DSM 28777T = KCTC33550T). An emended description of the genus Jeotgalibacillus is also provided.