- Volume 62, Issue Pt_5, 2012

A Gram-positive and aerobic bacterium, designated strain YIM 49065T, was isolated from rhizospheric soil of Jatropha curcas in Yunnan, China. This isolate formed branched and fragmented mycelia containing ll-diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan. The major cellular fatty acid profile was characterized by iso-C14 : 0, iso-C15 : 0 and iso-C16 : 0, and the predominant menaquinone was MK-8(H4). The DNA G+C content was 69.6 mol%. A phylogenetic analysis based on 16S rRNA gene sequence demonstrated that strain YIM 49065T was closely associated with Intrasporangium calvum DSM 43043T and Humihabitans oryzae KV-657T, exhibiting 98.8 % and 98.6 % 16S rRNA gene sequence similarities, respectively. Furthermore, DNA–DNA hybridizations among strains YIM 49065T, I. calvum DSM 43043T and H. oryzae DSM 22331T indicated that this isolate represented a novel species in the genus Intrasporangium . On the basis of these data, we proposed one novel species, Intrasporangium mesophilum sp. nov., for strain YIM 49065T ( = DSM 23217T  = CCTCC AA 209077T). Additionally, the comparison of biochemical and phylogenetic characters supported the reclassification of Humihabitans oryzae as a separate species within the genus Intrasporangium , Intrasporangium oryzae comb. nov. (type strain KV-657T  = JCM 15924T  = NBRC 101802T  = NRRL B-24470T).

A Gram-positive, strictly aerobic, non-spore-forming, irregular short rod, strain CAU 9625T, was isolated from a sediment of the Yellow Sea in the Republic of Korea. Strain CAU 9625T grew optimally at 37 °C, at pH 8.0 and in the presence of 1 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain CAU 9625T belonged to the genus Gulosibacter , which has one known member, Gulosibacter molinativorax . Strain CAU 9625T and G. molinativorax ON4T shared 97.8 % 16S rRNA gene sequence similarity and formed a distinct cluster (99 % bootstrap support) within the family Microbacteriaceae . DNA–DNA relatedness between strain CAU 9625T and G. molinativorax DSM 13485T was 35.4±0.9 %. The predominant menaquinone was MK-9. The major whole-cell sugars were ribose and glucose. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, an unidentified phospholipid and an unidentified lipid. The fatty acid composition was similar to that of G. molinativorax DSM 13485T, with anteiso-C15 : 0 as the predominant fatty acid. The DNA G+C content of strain CAU 9625T was 66.2 mol%. The phylogenetic and genetic distinctiveness and several differentiating phenotypic and chemotaxonomic properties revealed that strain CAU 9625T was distinguishable from G. molinativorax and other phylogenetic neighbours. On the basis of these data, strain CAU 9625T represents a novel species of the genus Gulosibacter , for which the name Gulosibacter chungangensis sp. nov. is proposed. The type strain is CAU 9625T ( = KCTC 13959T  = CCUG 60841T).

An actinomycete, designated XMU 198T, was isolated from the rhizosphere soil of a pharmaceutical plant, Typhonium giganteum Engl., collected in Xiamen City, China. 16S rRNA gene sequence analysis showed that the isolate exhibited highest sequence similarities with Kribbella flavida KACC 20148T, K. karoonensis Q41T and K. alba YIM 31975T (98.7, 98.4 and 98.2 %, respectively). The chemotaxonomic characteristics further supported the assignment of strain XMU 198T to the genus Kribbella : ll-diaminopimelic acid in the cell-wall peptidoglycan; glucose and galactose with minor amounts of ribose as the whole-cell sugars; polar lipids comprising phosphatidylglycerol, diphosphatidylglycerol, phosphatidylcholine, phosphatidylinositol and unidentified phospholipids; a fatty acid profile characterized by the predominance of iso-C16 : 0, iso-C14 : 0 and anteiso-C15 : 0; and MK-9(H4) as the main menaquinone. Gyrase subunit B gene (gyrB) sequence analysis showed that the genetic distances between strain XMU 198T and all other members of the genus Kribbella were greater than 0.014, the value used as the threshold for species delineation within this genus. A wide range of genotypic and phenotypic characteristics, as well as DNA–DNA relatedness between strain XMU 198T and K. flavida DSM 17836T (41.18 %), K. karoonensis Q41T (38.02 %) and K. alba DSM 15500T (50.58 %), distinguished the isolate from its closest phylogenetic neighbours. On the basis of the above data, a novel species of the genus Kribbella , Kribbella amoyensis sp. nov., is proposed. The type strain is XMU 198T ( = DSM 24683T = NBRC 107914T).

In this study, the taxonomic position of an actinobacterium, strain YIM M 10931T, which was isolated from a mangrove sediment sample collected in Dugong Creek, Little Andaman, India, was determined by a polyphasic approach. This Gram-positive, aerobic strain produced branched substrate mycelium and aerial hyphae, which differentiated into short, hooked or spiral spore chains. The organism contained meso-diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan. The whole cell sugars consisted of mannose, ribose, glucose, galactose and madurose. The cellular fatty acid profile mainly consisted of iso-C16 : 0, 10-methyl C18 : 0 and C16 : 0. The quinone system was predominantly composed of MK-9(H8) (45.5 %) and MK-9(H6) (39 %). The phospholipids detected were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol mannoside, phosphatidylinositol and two unknown phospholipids. The organism showed a combination of morphological and chemotaxonomic properties typical of members of the genus Actinomadura . Moreover, phylogenetic analysis based on a 16S rRNA gene sequence generated from the strain identified its closest relatives as Actinomadura cremea DSM 43676T (98.4 % sequence similarity), Actinomadura rifamycini DSM 43936T (97.4 %) and Actinomadura apis IM17-1T (96.9 %). It was obvious from the resulting phylogenetic trees that strain YIM M 10931T belongs to a distinct subclade within the evolutionary radiation of the genus Actinomadura . DNA–DNA hybridizations of strain YIM M 10931T with A. cremea DSM 43676T and A. rifamycini DSM 43936T were performed and further confirmed that the isolate represents a separate genomic species. Based on the phenotypic and genotypic characteristics presented, it is proposed that strain YIM M 10931T represents a novel species within the genus Actinomadura , for which the name Actinomadura sediminis sp. nov. is proposed; the type strain is YIM M 10931T ( = CCTCC AA 2010009T = DSM 45500T).

A Gram-stain-positive actinobacterium, strain PM267T, was isolated from a marine sediment sample in the Mediterranean Sea. The novel strain produced extensively branched substrate and aerial hyphae that carried spiral spore chains. Substrate and aerial mycelia were cream–white and white, respectively. Diffusible pigments were not observed. 16S rRNA gene sequence analysis revealed that strain PM267T belonged to the genus Streptomyces and shared a gene sequence similarity of 97.1 % with Streptomyces artemisiae YIM 63135T and Streptomyces armeniacus JCM 3070T. Values <97 % were obtained with other sequences representing members of the genus Streptomyces . The cell wall peptidoglycan contained ll -diaminopimelic acid. MK-9(H8) was the major menaquinone. The phospholipid pattern included phosphatidylethanolamine as diagnostic lipid (type II). Major fatty acids found were iso- and anteiso- fatty acids. The G+C content of the DNA was 71.2 mol%. The strain was halotolerant and was able to grow in the presence of 9 % (w/v) NaCl (with an optimum of 2 %). On the basis of these results and additional physiological data obtained in the present study, strain PM267T represents a novel species within the genus Streptomyces for which the name Streptomyces pharmamarensis sp. nov. is proposed (type strain PM267T  = CECT 7841T  = DSM 42032T).

Strain MJ31T, a Gram-reaction-positive, aerobic, rod-shaped, non-motile bacterium, was isolated from a sludge sample collected at the Daejeon sewage-disposal plant, in South Korea, and characterized in order to determine its taxonomic position. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain MJ31T belonged to the genus Nocardioides , appearing most closely related to Nocardioides dubius KSL-104T (98.6 % sequence similarity), Nocardioides jensenii DSM 20641T (97.6 %), Nocardioides daedukensis MDN22T (97.2 %) and Nocardioides mesophilus MSL-22T (97.0 %). The chemotaxonomic properties of strain MJ31T were consistent with those of the genus Nocardioides : MK-8(H4) was the predominant menaquinone, iso-C16 : 0, iso-C17 : 0 and C18 : 1ω9c were the predominant cellular fatty acids, and the cell-wall peptidoglycan was based on ll-2,6-diaminopimelic acid. The genomic DNA G+C content of strain MJ31T was 71.2 mol%. Some differential phenotypic properties and low DNA–DNA relatedness values (<28 %) with the type strains of closely related species indicated that strain MJ31T represents a novel species, for which the name Nocardioides daejeonensis sp. nov. is proposed. The type strain is MJ31T ( = KCTC 19772T = JCM 16922T).

A Gram-positive, acid-fast and rapidly growing rod, designated F4T, was isolated from a soil sample of Haikou in China. The isolate shared 98.2 % 16S rRNA gene sequence similarity with Mycobacterium monacense B9-21-178T, 96.2 % hsp65 sequence similarity with M. monacense FI-05352 and 79.6 % 16S–23S rRNA internal transcribed spacer sequence similarity with M. monacense B9-21-178T. DNA–DNA relatedness between the isolate and M. monacense DSM 44395T was 43.5 %. The morphological analysis and physiological tests also showed that the isolate differed from any strain reported to date. The mycolic acid profile and the cellular fatty acid composition were also determined. On the basis of phenotypic and chemotaxonomic characteristics and phylogenetic data, it was concluded that strain F4T ( = CGMCC 4.5724T = JCM 17423T) merited classification as the type strain of a novel species, for which the name Mycobacterium litorale sp. nov. is proposed.

A novel moderately halophilic bacterium, designated strain K170T, was isolated from Keke Salt Lake in Qinghai, China. The strain grew with 0–22 % (w/v) NaCl, at 4–50 °C and at pH 6–11, with optimum growth in 3 % (w/v) NaCl, at 40 °C and at pH 8. The predominant respiratory quinone was menaquinone 7 (MK-7). The polar lipids included diphosphatidylglycerol, phosphatidylglycerol, unidentified phospholipids, aminolipids and glycolipids. The major cellular fatty acids were anteiso-C15 : 0, iso-C15 : 0 and anteiso-C17 : 0. The DNA G+C content was 35.8 mol%. Phylogenetic analysis based on the full-length 16S rRNA gene sequence revealed that strain K170T was a member of the genus Gracilibacillus . High levels of 16S rRNA gene sequence similarity were found between strain K170T and Gracilibacillus boraciitolerans DSM 17256T (97.3 %) and Gracilibacillus thailandensis JCM 15569T (97.1 %). 16S rRNA gene sequence similarities between strain K170T and the type strains of other recognized members of the genus Gracilibacillus were below 97 %. The DNA–DNA hybridization values of strain K170T with G. boraciitolerans DSM 17256T and G. thailandensis JCM 15569T were 21.9 % and 34.3 %, respectively. On the basis of these results, strain K170T is considered to represent a novel species of the genus Gracilibacillus , for which the name Gracilibacillus kekensis sp. nov. is proposed; the type strain is K170T ( = CGMCC 1.10681T = DSM 23178T).

Three Gram-staining-positive, strictly aerobic, motile, catalase-positive, endospore-forming rods, designated WCC 4582T, WCC 4581 and WCC 4583, were isolated from two different food sources and a pharmaceuticals production site. The three isolates were highly similar in their 16S rRNA gene sequences (100 % similarity) and groEL sequences (99.2–100 % similarity), Fourier-transform infrared spectroscopic fingerprints and other features tested. The isolates were most closely related to Bacillus horneckiae ; the isolates and the type strain of B. horneckiae shared 97.6 % and 89.6 % 16S rRNA gene and groEL sequence similarities, respectively. The organisms grew optimally at 30 °C, at pH 7 and in the presence of 0.5 % (w/v) NaCl. The cell-wall peptidoglycan of WCC 4582T contained meso-diaminopimelic acid (A1γ) and the genomic DNA G+C content was 36.4 mol%. DNA–DNA relatedness between strain WCC 4582T and B. horneckiae NRRL B-59162T was 17 %. The three isolates are considered to represent a novel species of the genus Bacillus , for which the name Bacillus kochii sp. nov. is proposed. The type strain is WCC 4582T ( = DSM 23667T = CCUG 59877T = LMG 25855T).

A Gram-staining-positive, non-motile and non-spore-forming lactic acid bacterium, designated strain M2T, was isolated from fermented brown algae (Undaria pinnatifida) kimchi in South Korea. Cells of the isolate were facultatively anaerobic ovoids and showed catalase- and oxidase-negative reactions. Growth of strain M2T was observed at 4–35 °C and at pH 5.0–9.0. The G+C content of the genomic DNA was 42.5 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain M2T belonged to the genus Leuconostoc and was most closely related to Leuconostoc inhae IH003T, Leuconostoc kimchii IH25T, Leuconostoc gasicomitatum LMG 18811T, Leuconostoc gelidum DSM 5578T, Leuconostoc palmae TMW2.694T and Leuconostoc holzapfelii BFE 7000T with 98.9 %, 98.8 %, 98.8 %, 98.7 %, 98.5 % and 98.2 % sequence similarity, respectively. DNA–DNA hybridization values between strain M2T and Leuconostoc inhae KACC 12281T, Leuconostoc kimchii IH25T, Leuconostoc gelidum KACC 12256T, Leuconostoc gasicomitatum KACC 13854T, Leuconostoc palmae DSM 21144T and Leuconostoc holzapfelii DSM 21478T were 13.8±3.2 %, 14.3±3.4 %, 9.9±1.0 %, 13.2±0.8 %, 22.4±4.9 % and 16.2±4.6 %, respectively, which allowed differentiation of strain M2T from the closely related species of the genus Leuconostoc . On the basis of phenotypic and molecular properties, strain M2T represents a novel species in the genus Leuconostoc , for which the name Leuconostoc miyukkimchii sp. nov. is proposed. The type strain is M2T ( = KACC 15353T  = JCM 17445T).

‘Bacillus macroides’ ATCC 12905T ( = DSM 54T = LMG 18474T), isolated in 1947 from cow dung, was not included in the Approved Lists of Bacterial Names and so it lost standing in bacteriological nomenclature. Reinvestigation of the strain, including DNA–DNA relatedness experiments, revealed that ‘Bacillus macroides’ is genomically distinct from its closest relatives Lysinibacillus xylanilyticus , Lysinibacillus boronitolerans and Lysinibacillus fusiformis (as determined by 16S rRNA gene sequence analysis, with pairwise similarity values of 99.2, 98.8 and 98.5 %, respectively, with the type strains of these species). Further analysis showed that ‘Bacillus macroides’ shares the A4α l-Lys–d-Asp peptidoglycan type with other members of the genus Lysinibacillus and can thus be attributed to this genus. These results, combined with additional phenotypic data, justify the description of strain LMG 18474T ( = DSM 54T = ATCC 12905T) as Lysinibacillus macroides sp. nov., nom. rev.

Two strains of lactic acid bacteria, designated SU 18T and SU 83, were isolated from silage prepared with Sudan grass [Sorghum sudanense (Piper) Stapf.]. The isolates were Gram-stain-positive, catalase-negative, facultatively anaerobic rods that did not produce gas from glucose. The isolates exhibited ≥93.5 % DNA–DNA relatedness to each other and shared the same phenotypic characteristics, which indicated that they belonged to a single species. The DNA G+C content was 58.5–59.2 mol%. On the basis of 16S rRNA gene sequence analysis, the isolates were placed in the genus Lactobacillus . Their closest phylogenetic neighbours were Lactobacillus manihotivorans JCM 12514T and Lactobacillus camelliae JCM 13995T (95.9 and 96.8 % 16S rRNA gene sequence similarity, respectively, with strain SU 18T). Ribotyping revealed that strain SU 18T was well separated from L. manihotivorans JCM 12514T and L. camelliae JCM 13995T. Strain SU 18T exhibited ≤23.7 % DNA–DNA relatedness with its closest phylogenetic neighbours. The isolates represent a novel species in the genus Lactobacillus , for which the name Lactobacillus nasuensis sp. nov. is proposed. The type strain is SU 18T ( = JCM 17158T  = CGMCC 1.10801T). The description of the genus Lactobacillus is also amended.

Two novel Gram-stain-negative, aerobic strains (ZXM100T and ZXM098) were isolated from seawater in the coastal region of Qingdao (36.027° N 120.184° E), China, during a massive green algae bloom. Cells were ovoid to irregular short rods and lacked flagella. Poly-β-hydroxybutyrate was accumulated. Cells did not contain bacteriochlorophyll a. The isolates grew at NaCl concentrations of 3–9 % and 22–28 °C. C18 : 1ω7c, C18 : 0 and C16 : 0 were the major fatty acid components. The polar lipids of ZXM100T were phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, a single lipid and an unidentified aminolipid. The predominant isoprenoid quinone of ZXM100T was Q-10. Phylogenetic analysis based on 16S rRNA gene sequences showed that strains belonged to the family Rhodobacteraceae . Based on the results obtained in this study, strain ZXM100T was considered to represent a novel species in a new genus, for which the name Lentibacter algarum gen. nov., sp. nov. is proposed. The type strain is ZXM100T ( = LMG 24861T = CGMCC 1.10234T); the DNA G+C content of the type strain is 54.6 mol%.

A yellow-pigmented bacterial strain designated TNR-25T was isolated from spring water in Taiwan and was characterized using a polyphasic taxonomic approach. Strain TNR-25T was Gram-negative, obligately aerobic, rod-shaped, non-motile and non-spore-forming. Growth occurred at 15–40 °C (optimum, 25 °C), at pH 6.0–10.0 (optimum, pH 7.0) and with 0–0.5 % NaCl (optimum, 0 %). Phylogenetic analyses based on 16S rRNA gene sequences showed that strain TNR-25T belonged to the genus Inhella and its closest neighbour was Inhella inkyongensis IMCC1713T with 98.1 % sequence similarity. The major fatty acids (>10 %) of strain TNR-25T were summed feature 3 (comprising C16 : 1ω7c and/or C16 : 1ω6c) and C16 : 0. The major cellular hydroxy fatty acids were C10 : 0 3-OH and C12 : 0 3-OH. The isoprenoid quinone was Q-8 and the DNA G+C content was 69.6 mol%. The polar lipid profile consisted of a mixture of phosphatidylethanolamine, phosphatidylglycerol, phosphatidylserine, diphosphatidylglycerol and several uncharacterized phospholipids. The DNA–DNA relatedness between strain TNR-25T and I. inkyongensis IMCC1713T was about 30.6–35.5 %. On the basis of the genotypic and phenotypic data, strain TNR-25T represents a novel species in the genus Inhella , for which the name Inhella fonticola sp. nov. is proposed; the type strain is TNR-25T ( = BCRC 80211T = LMG 25721T).

Two strains (JA492T and JA590) of spiral-shaped, anaerobic, Gram-stain-negative, motile, purple non-sulfur bacteria were isolated from aquatic sediments from a bird sanctuary and a stream, respectively, and were characterized by a polyphasic taxonomic approach. Bacteriochlorophyll a and carotenoids (rhodopin, lycopene, hydroxylycopene glucoside and dihydroxylycopene diglucoside) were present as photosynthetic pigments. Intracellular photosynthetic membranes were of the stacked type. The major fatty acids were C18 : 1ω7c, C16 : 0 and summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c) in both strains. Ubiquinones and menaquinones were present as major quinone components. The genomic DNA G+C contents of strains JA492T and JA590 were 63.8 and 61.5 mol%, respectively. Both strains were closely related (mean DNA–DNA hybridization >70 %). Phylogenetic analysis showed that the strains clustered with species of the genus Phaeospirillum of the family Rhodospirillaceae , class Alphaproteobacteria . Based on 16S rRNA gene sequence analysis, both strains showed highest sequence similarity with Phaeospirillum oryzae JA317T (97.2–97.4 %), Phaeospirillum molischianum DSM 120T (96.5–96.7 %), Phaeospirillum fulvum DSM 113T (96.7–96.9 %) and Phaeospirillum chandramohanii JA145T (96.5–96.7 %). DNA–DNA relatedness between strain JA492T and its closest relative in the genus Phaeospirillum was less than 42 %. It is evident from phenotypic, chemotaxonomic and molecular genetic data that strain JA492T represents a novel species of the genus Phaeospirillum , for which the name Phaeospirillum tilakii sp. nov. is proposed; the type strain is JA492T ( = NBRC 107650T = KCTC 15012T).

A facultatively anaerobic, chitinolytic bacterium, strain KL-9T, was isolated from a freshwater lake in Taiwan and characterized by using a polyphasic taxonomic approach. Cells of strain KL-9T were Gram-negative, rod-shaped, motile by means of a single polar flagellum and non-spore-forming. Growth occurred at 15–40 °C (optimum, 30–37 °C), at pH 7.0–9.0 (optimum, pH 8.0) and with 0–1.0 % NaCl (optimum, 0 %). The predominant fatty acids were summed feature 3 (comprising C16 : 1ω7c and/or C16 : 1ω6c) and C16 : 0. The major isoprenoid quinone was Q-8. The DNA G+C content of strain KL-9T was 64.6 mol%. The polar lipid profile consisted of a mixture of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidyldimethylethanolamine and several uncharacterized phospholipids and aminolipids. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain KL-9T formed a distinct lineage with respect to closely related genera within the class Betaproteobacteria , being most closely related to members of the genera Leeia , Chitinimonas , Silvimonas and Andreprevotia . Levels of 16S rRNA gene sequence similarity with respect to the type strains of type species of these genera were below 91 %. On the basis of genotypic and phenotypic data, strain KL-9T is thus considered to represent a novel species of a new genus within the class Betaproteobacteria , for which the name Chitinivorax tropicus gen. nov., sp. nov. is proposed. The type strain of Chitinivorax tropicus is KL-9T ( = BCRC 80168T = LMG 25530T).

A bacterial strain designated antisso-13T was isolated from a salt pan in southern Taiwan and characterized using a polyphasic taxonomic approach. Strain antisso-13T was Gram-negative, aerobic, creamy white in colour, rod-shaped and motile by single monopolar flagellum. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain antisso-13T belonged to the genus Neptuniibacter and its closest neighbour was Neptuniibacter caesariensis MED92T, with 96.7 % sequence similarity. Phylogenetic analyses based on rpoB and recA gene sequences and deduced amino acid sequences also revealed that the novel strain and N. caesariensis MED92T formed a distinct phylogenetic cluster. Strain antisso-13T exhibited optimal growth at 25–30 °C, with 2.0–4.0 % NaCl and at pH 7.0. Cellular fatty acids were C16 : 1ω7c (39.8 %), C18 : 1ω7c (29.4 %), C16 : 0 (15.5 %), C10 : 0 3-OH (7.5 %), anteiso-C11 : 0 (1.8 %), C18 : 0 (1.8 %), C11 : 0 2-OH (1.6 %), iso-C10 : 0 (1.2 %) and C14 : 0 (1.1 %). The major respiratory quinone was ubiquinone Q-8. The polar lipid profile consisted of a mixture of phosphatidylglycerol, phosphatidylethanolamine and several uncharacterized polar lipids and the DNA G+C content was 54.2 mol%. The results of physiological and biochemical tests allowed clear phenotypic differentiation of this isolate from previously described members of the genus Neptuniibacter . It is evident from the genotypic and phenotypic data that strain antisso-13T should be classified as a representative of a novel species in the genus Neptuniibacter . The name proposed for this taxon is Neptuniibacter halophilus sp. nov.; the type strain is antisso-13T ( = LMG 25378T = BCRC 80079T). An emended description of the genus Neptuniibacter is provided.